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pubmed23n0812_12689

Cochlear and brainstem audiologic findings in normal hearing tinnitus subjects in comparison with non-tinnitus control group.

While most tinnitus cases have some degree of hearing impairment, a small percent of the patients admitted to Ear, Nose and Throat Clinics or Hearing Evaluation Centers are those who complain of tinnitus despite having normal hearing thresholds. Present study was performed in order to better understanding of the probable causes of tinnitus and to investigate possible changes in the cochlear and auditory brainstem function in normal hearing patients with chronic tinnitus. Altogether, 63 ears (31 ears with tinnitus and 32 ears without tinnitus) were examined. The prevalence of transient evoked otoacoustic emissions and characteristics of the auditory brainstem response components including wave latencies and wave amplitudes was determined in the two groups and analyzed with appropriate statistical methods. There was no difference between the prevalence of transient evoked emissions in the two groups. The mean difference between absolute latencies of waves I, III and V was less than 0.1 ms between the two groups that were not statistically significant. Also, the interpeak latency values of I-III, III-V and I-V in both groups had no significant difference. Only the V/I amplitude ratio in the tinnitus group was significantly larger than the other group (p =0.04). The changes observed in amplitude of waves, especially in the later ones, can be considered as an Audiologic finding in normal hearing tinnitus subjects and its possible role in generation of tinnitus in these patients must be investigated.

Cochlear and brainstem audiologic findings in normal hearing tinnitus subjects in comparison with non-tinnitus control group. While most tinnitus cases have some degree of hearing impairment, a small percent of the patients admitted to Ear, Nose and Throat Clinics or Hearing Evaluation Centers are those who complain of tinnitus despite having normal hearing thresholds. Present study was performed in order to better understanding of the probable causes of tinnitus and to investigate possible changes in the cochlear and auditory brainstem function in normal hearing patients with chronic tinnitus. Altogether, 63 ears (31 ears with tinnitus and 32 ears without tinnitus) were examined. The prevalence of transient evoked otoacoustic emissions and characteristics of the auditory brainstem response components including wave latencies and wave amplitudes was determined in the two groups and analyzed with appropriate statistical methods. There was no difference between the prevalence of transient evoked emissions in the two groups. The mean difference between absolute latencies of waves I, III and V was less than 0.1 ms between the two groups that were not statistically significant. Also, the interpeak latency values of I-III, III-V and I-V in both groups had no significant difference. Only the V/I amplitude ratio in the tinnitus group was significantly larger than the other group (p =0.04). The changes observed in amplitude of waves, especially in the later ones, can be considered as an Audiologic finding in normal hearing tinnitus subjects and its possible role in generation of tinnitus in these patients must be investigated.

pubmed23n0864_10403

WAY208466 inhibits glutamate release at hippocampal nerve terminals.

Evidence suggests that the glutamatergic system plays a crucial role in the pathophysiology and treatment of depression. This study investigates the effect of WAY208466, a 5-HT6 receptor agonist exhibiting an antidepressant effect, on glutamate release from rat hippocampal nerve terminals (synaptosomes). WAY208466 inhibited the Ca(2+)-dependent release of glutamate that was evoked by exposing the synaptosomes to the potassium channel blocker 4-aminopyridine, and the selective 5-HT6 receptor antagonist SB258585 blocked this phenomenon. The WAY208466-mediated inhibition of glutamate release was associated with a reduction of 4-aminopyridine-induced increase in the cytosolic free Ca(2+) concentration ([Ca(2+)]C) mediated via Cav2.2 (N-type) and Cav2.1 (P/Q-type) channels. WAY208466 did not alter the resting synaptosomal membrane potential or 4-aminopyridine-mediated depolarization; thus, the inhibition of the Ca(2+) influx could not be attributed to the decrease in synaptosomal excitability caused by 5-HT6 receptor activation. Furthermore, the effect of WAY208466 on 4-aminopyridine-evoked glutamate release was prevented by a Gi/Go-protein inhibitor pertussis toxin, adenylate cyclase inhibitor SQ22536, and a protein kinase A inhibitor H89. These results suggest that WAY208466 acts at the 5-HT6 receptors present in the hippocampal nerve terminals to suppress the Gi/Go-protein-coupled adenylate cyclase/protein kinase A cascade, which subsequently reduces the Ca(2+) influx via N- and P/Q-type Ca(2+) channels to inhibit the evoked glutamate release. This finding implicated a potential therapeutic role of 5-HT6 receptor agonist in the treatment of depression and other neurological diseases associated with glutamate excitotoxicity.

WAY208466 inhibits glutamate release at hippocampal nerve terminals. Evidence suggests that the glutamatergic system plays a crucial role in the pathophysiology and treatment of depression. This study investigates the effect of WAY208466, a 5-HT6 receptor agonist exhibiting an antidepressant effect, on glutamate release from rat hippocampal nerve terminals (synaptosomes). WAY208466 inhibited the Ca(2+)-dependent release of glutamate that was evoked by exposing the synaptosomes to the potassium channel blocker 4-aminopyridine, and the selective 5-HT6 receptor antagonist SB258585 blocked this phenomenon. The WAY208466-mediated inhibition of glutamate release was associated with a reduction of 4-aminopyridine-induced increase in the cytosolic free Ca(2+) concentration ([Ca(2+)]C) mediated via Cav2.2 (N-type) and Cav2.1 (P/Q-type) channels. WAY208466 did not alter the resting synaptosomal membrane potential or 4-aminopyridine-mediated depolarization; thus, the inhibition of the Ca(2+) influx could not be attributed to the decrease in synaptosomal excitability caused by 5-HT6 receptor activation. Furthermore, the effect of WAY208466 on 4-aminopyridine-evoked glutamate release was prevented by a Gi/Go-protein inhibitor pertussis toxin, adenylate cyclase inhibitor SQ22536, and a protein kinase A inhibitor H89. These results suggest that WAY208466 acts at the 5-HT6 receptors present in the hippocampal nerve terminals to suppress the Gi/Go-protein-coupled adenylate cyclase/protein kinase A cascade, which subsequently reduces the Ca(2+) influx via N- and P/Q-type Ca(2+) channels to inhibit the evoked glutamate release. This finding implicated a potential therapeutic role of 5-HT6 receptor agonist in the treatment of depression and other neurological diseases associated with glutamate excitotoxicity.

pubmed23n0376_3611

The impact of the thermal sensitivity of cytochrome c oxidase on the respiration rate of Arctic charr red muscle mitochondria. [email protected].

To assess if cytochrome c oxidase could determine the response of mitochondrial respiration to changes in environmental temperature in ectotherms, we performed KCN titration of the respiration rate and cytochrome c oxidase activity in mitochondria from Arctic charr (Salvelinusfontinalis) muscle at four different temperatures (1 degrees C, 6 degrees C, 12 degrees C, and 18 degrees C). Our data showed an excess of cytochrome c oxidase activity over the mitochondrial state 3 respiration rate. Mitochondrial oxygen consumption rates reached approximately 12% of the cytochrome c oxidase maximal capacity at every temperature. Also, following titration, the mitochondrial respiration rate significantly decreased when KCN reached concentrations that inhibit almost 90% of the cytochrome c oxidase activity. This strongly supports the idea that the thermal sensitivity of the maximal mitochondrial respiration rate cannot be dictated by the effect of temperature on cytochrome c oxidase catalytic capacity. Furthermore, the strong similarity of the Q10s of mitochondrial respiration and cytochrome c oxidase activity suggests a functional or structural link between the two. The functional link could be coevolution of parts of the mitochondrial system to maintain optimal functions in most of the temperature range encountered by organisms.

The impact of the thermal sensitivity of cytochrome c oxidase on the respiration rate of Arctic charr red muscle mitochondria. [email protected]. To assess if cytochrome c oxidase could determine the response of mitochondrial respiration to changes in environmental temperature in ectotherms, we performed KCN titration of the respiration rate and cytochrome c oxidase activity in mitochondria from Arctic charr (Salvelinusfontinalis) muscle at four different temperatures (1 degrees C, 6 degrees C, 12 degrees C, and 18 degrees C). Our data showed an excess of cytochrome c oxidase activity over the mitochondrial state 3 respiration rate. Mitochondrial oxygen consumption rates reached approximately 12% of the cytochrome c oxidase maximal capacity at every temperature. Also, following titration, the mitochondrial respiration rate significantly decreased when KCN reached concentrations that inhibit almost 90% of the cytochrome c oxidase activity. This strongly supports the idea that the thermal sensitivity of the maximal mitochondrial respiration rate cannot be dictated by the effect of temperature on cytochrome c oxidase catalytic capacity. Furthermore, the strong similarity of the Q10s of mitochondrial respiration and cytochrome c oxidase activity suggests a functional or structural link between the two. The functional link could be coevolution of parts of the mitochondrial system to maintain optimal functions in most of the temperature range encountered by organisms.

pubmed23n1052_8587

Pulmonary Sparganosis: Tunnel Sign and Migrating Sign on Computed Tomography.

A 77-year-old woman presented at our hospital to undergo a close examination of an abnormal shadow which was observed on a chest radiograph. Contrast-enhanced computed tomography (CT) images in the lung window revealed a tortuous tunnel structure (tunnel sign), which was suspected to be the migration path of a parasite. Furthermore, CT images in the mediastinal window showed a linear filling defect from the right inferior pulmonary vein to the venous ostium in the left atrium (migrating sign), which was suspected to be a migrating parasite in the pulmonary vein. Tunnel and migrating signs on chest CT images were helpful in diagnosing pulmonary sparganosis.

Pulmonary Sparganosis: Tunnel Sign and Migrating Sign on Computed Tomography. A 77-year-old woman presented at our hospital to undergo a close examination of an abnormal shadow which was observed on a chest radiograph. Contrast-enhanced computed tomography (CT) images in the lung window revealed a tortuous tunnel structure (tunnel sign), which was suspected to be the migration path of a parasite. Furthermore, CT images in the mediastinal window showed a linear filling defect from the right inferior pulmonary vein to the venous ostium in the left atrium (migrating sign), which was suspected to be a migrating parasite in the pulmonary vein. Tunnel and migrating signs on chest CT images were helpful in diagnosing pulmonary sparganosis.

pubmed23n0571_7204

The ester group: how hydrofluoroalkane-philic is it?

Pressurized metered-dose inhalers (pMDIs) have been recognized as potential devices for the delivery of systemically acting drugs, including biomolecules, to and through the lungs. Therefore, the development of novel excipients capable of imparting stability to suspension formulations in hydrofluoroalkane (HFA) propellants is of great relevance because many of the drugs of interest are poorly soluble in HFAs. In this work, we use ab initio calculations and chemical force microscopy (CFM) to determine the HFA-philicity of the biodegradable and biocompatible ester moiety quantitatively. The complementary information obtained from the binding energy calculations and adhesion force measurements are used to gain microscopic insight into the relationship between the chemistry of the moiety of interest and its solvation in HFA. A lactide (LA)-based copolymer surfactant was synthesized and characterized, and its ability to stabilize a dispersion of micronized budesonide in HFA227 was demonstrated. These results corroborate the ab initio calculations and CFM and show that the LA-based moiety is a suitable candidate for enhancing the stability of dispersions in HFA-based pMDIs.

The ester group: how hydrofluoroalkane-philic is it? Pressurized metered-dose inhalers (pMDIs) have been recognized as potential devices for the delivery of systemically acting drugs, including biomolecules, to and through the lungs. Therefore, the development of novel excipients capable of imparting stability to suspension formulations in hydrofluoroalkane (HFA) propellants is of great relevance because many of the drugs of interest are poorly soluble in HFAs. In this work, we use ab initio calculations and chemical force microscopy (CFM) to determine the HFA-philicity of the biodegradable and biocompatible ester moiety quantitatively. The complementary information obtained from the binding energy calculations and adhesion force measurements are used to gain microscopic insight into the relationship between the chemistry of the moiety of interest and its solvation in HFA. A lactide (LA)-based copolymer surfactant was synthesized and characterized, and its ability to stabilize a dispersion of micronized budesonide in HFA227 was demonstrated. These results corroborate the ab initio calculations and CFM and show that the LA-based moiety is a suitable candidate for enhancing the stability of dispersions in HFA-based pMDIs.

pubmed23n0836_18620

Correlation of Real Time PCR Cycle Threshold Cut-Off with Bordetella pertussis Clinical Severity.

Bordetella pertussis testing performed using real-time polymerase chain reaction (RT-PCR) is interpreted based on a cycle threshold (Ct) value. At Public Health Ontario Laboratories (PHOL), a Ct value <36 is reported as positive, and Ct values ≥36 and <40 are reported as indeterminate. PHOL reported indeterminate results to physicians and public health units until May 2012, after which these results were only reported to physicians. We investigated the association between Ct value and disease symptom and severity to examine the significance of indeterminate results clinically, epidemiologically and for public health reporting. B. pertussis positive and indeterminate RT-PCR results were linked to pertussis cases reported in the provincial Integrated Public Health Information System (iPHIS), using deterministic linkage. Patients with positive RT-PCR results had a lower median age of 10.8 years compared to 12.0 years for patients with indeterminate results (p = 0.24). Hospitalized patients had significantly lower Ct values than non-hospitalized patients (median Ct values of 20.7 vs. 31.6, p<0.001). The proportion of patients reporting the most indicative symptoms of pertussis did not differ between patients with positive vs. indeterminate RT-PCR results. Taking the most indicative symptoms of pertussis as the gold-standard, the positive predictive value of the RT-PCR test was 68.1%. RT-PCR test results should be interpreted in the context of the clinical symptoms, age, vaccination status, prevalence, and other factors. Further information on interpretation of indeterminate RT-PCR results may be needed, and the utility of reporting to public health practitioners should be re-evaluated.

Correlation of Real Time PCR Cycle Threshold Cut-Off with Bordetella pertussis Clinical Severity. Bordetella pertussis testing performed using real-time polymerase chain reaction (RT-PCR) is interpreted based on a cycle threshold (Ct) value. At Public Health Ontario Laboratories (PHOL), a Ct value <36 is reported as positive, and Ct values ≥36 and <40 are reported as indeterminate. PHOL reported indeterminate results to physicians and public health units until May 2012, after which these results were only reported to physicians. We investigated the association between Ct value and disease symptom and severity to examine the significance of indeterminate results clinically, epidemiologically and for public health reporting. B. pertussis positive and indeterminate RT-PCR results were linked to pertussis cases reported in the provincial Integrated Public Health Information System (iPHIS), using deterministic linkage. Patients with positive RT-PCR results had a lower median age of 10.8 years compared to 12.0 years for patients with indeterminate results (p = 0.24). Hospitalized patients had significantly lower Ct values than non-hospitalized patients (median Ct values of 20.7 vs. 31.6, p<0.001). The proportion of patients reporting the most indicative symptoms of pertussis did not differ between patients with positive vs. indeterminate RT-PCR results. Taking the most indicative symptoms of pertussis as the gold-standard, the positive predictive value of the RT-PCR test was 68.1%. RT-PCR test results should be interpreted in the context of the clinical symptoms, age, vaccination status, prevalence, and other factors. Further information on interpretation of indeterminate RT-PCR results may be needed, and the utility of reporting to public health practitioners should be re-evaluated.

pubmed23n0416_20567

Small peripheral pulmonary carcinomas evaluated with dynamic MR imaging: correlation with tumor vascularity and prognosis.

To correlate the findings at contrast material-enhanced dynamic magnetic resonance (MR) imaging of small peripheral pulmonary carcinomas with tumor vascularity and prognosis. Ninety-four patients with small peripheral pulmonary carcinomas who underwent surgical resection were examined retrospectively. Pathologic specimens were stained with hematoxylin-eosin and elastin-van Gieson. CD34 and vascular endothelial growth factor (VEGF) were assessed immunohistochemically. Delineated CD34-positive cells were counted as microvessels. Dynamic MR imaging was performed prior to and at 1, 2, 3, 4, 5, 6, and 8 minutes after injection of a bolus of gadopentetate dimeglumine. The two observers reviewed all images, and two pathologists performed all histologic analyses; a decision was determined with consensus. The maximum enhancement ratio (MER), the time lapse between the completion of the injection and the point of maximum signal intensity (Tmax), the washout ratio, and the slope value of the time-signal intensity curve were correlated with the microvessel density. VEGF-positive and VEGF-negative tumors were compared. All statistical analyses were performed by using nonparametric methods. The MER and the slope value were positively correlated, and the Tmax was negatively correlated (Spearman rank test, P <.0001, all comparisons) with the microvessel counts. The distribution of elastic and collagen fibers correlated with the washout ratio (Kruskal-Wallis test, P <.001). There was a statistically significant difference between the slope value of VEGF-positive tumors and that of VEGF-negative tumors (Mann-Whitney U test, P <.0001). Patients with VEGF-positive tumors had a significantly shorter overall survival than did those with VEGF-negative tumors (log-rank test, P <.0001). Dynamic MR imaging findings correlate with tumor vascularity and may be helpful in the prediction of prognosis.

Small peripheral pulmonary carcinomas evaluated with dynamic MR imaging: correlation with tumor vascularity and prognosis. To correlate the findings at contrast material-enhanced dynamic magnetic resonance (MR) imaging of small peripheral pulmonary carcinomas with tumor vascularity and prognosis. Ninety-four patients with small peripheral pulmonary carcinomas who underwent surgical resection were examined retrospectively. Pathologic specimens were stained with hematoxylin-eosin and elastin-van Gieson. CD34 and vascular endothelial growth factor (VEGF) were assessed immunohistochemically. Delineated CD34-positive cells were counted as microvessels. Dynamic MR imaging was performed prior to and at 1, 2, 3, 4, 5, 6, and 8 minutes after injection of a bolus of gadopentetate dimeglumine. The two observers reviewed all images, and two pathologists performed all histologic analyses; a decision was determined with consensus. The maximum enhancement ratio (MER), the time lapse between the completion of the injection and the point of maximum signal intensity (Tmax), the washout ratio, and the slope value of the time-signal intensity curve were correlated with the microvessel density. VEGF-positive and VEGF-negative tumors were compared. All statistical analyses were performed by using nonparametric methods. The MER and the slope value were positively correlated, and the Tmax was negatively correlated (Spearman rank test, P <.0001, all comparisons) with the microvessel counts. The distribution of elastic and collagen fibers correlated with the washout ratio (Kruskal-Wallis test, P <.001). There was a statistically significant difference between the slope value of VEGF-positive tumors and that of VEGF-negative tumors (Mann-Whitney U test, P <.0001). Patients with VEGF-positive tumors had a significantly shorter overall survival than did those with VEGF-negative tumors (log-rank test, P <.0001). Dynamic MR imaging findings correlate with tumor vascularity and may be helpful in the prediction of prognosis.

pubmed23n0492_4114

Adsorptive stripping voltammetric determination of vanadium as chloranilic acid complex.

Traces of vanadium(V) can be determined up to 21 ng/l by adsorptive stripping voltammetry using 2,5-dichloro-1,4-dihydroxy-3,6-benzoquinone (chloranilic acid) as complex forming reagent. The complex is stable in a 0.1 mol/l acetate buffer (pH 4.6) solution and adsorbed at the electrode surface in a potential range between -150 mV and -400 mV. The adsorption process is solved by alternating current (ac) voltammetric measurements. The method is applied to different real samples.

Adsorptive stripping voltammetric determination of vanadium as chloranilic acid complex. Traces of vanadium(V) can be determined up to 21 ng/l by adsorptive stripping voltammetry using 2,5-dichloro-1,4-dihydroxy-3,6-benzoquinone (chloranilic acid) as complex forming reagent. The complex is stable in a 0.1 mol/l acetate buffer (pH 4.6) solution and adsorbed at the electrode surface in a potential range between -150 mV and -400 mV. The adsorption process is solved by alternating current (ac) voltammetric measurements. The method is applied to different real samples.

pubmed23n0911_24194

The renoprotective effects of mannitol and udenafil in renal ischemia-reperfusion injury model.

The aim of this study was to investigate and compare the effects of udenafil and mannitol in an experimental renal ischemia-reperfusion (I/R) injury model. A total of 64 female Wister Albino rats were used. Right nephrectomy was performed in all groups. In the control group; I/R injury was not performed. In the I/R group; left renal pedicle was clamped for 45 minutes and then underwent 60 minutes and 24 hours of reperfusion. In the mannitol group; 1 mL 20% mannitol was given intravenously 15 minutes before clamping. In the udenafil group; 10-mg/kg udenafil was given orally 1 hour before clamping. Creatinine (Cr), blood urea nitrogen (BUN), Cr clearance, malondialdehyde, neutrophil gelatinase associated lipocalin (NGAL), histological examination and DNA damage (Comet Assay method) levels were compared in tissue, serum and urine samples. Udenafil had a better protective effect than mannitol according to biochemical parameters (Cr, BUN, Cr clearance, and NGAL levels) and histopathological findings when compared with the I/R group. In the Comet sampling analysis no significant difference was detected. Udenafil has a better renoprotective effect than mannitol against I/R injury and this effect supports more functional improvements. Further clinical trials are needed to demonstrate those effects and clinical utility of udenafil for that purpose in humans.

The renoprotective effects of mannitol and udenafil in renal ischemia-reperfusion injury model. The aim of this study was to investigate and compare the effects of udenafil and mannitol in an experimental renal ischemia-reperfusion (I/R) injury model. A total of 64 female Wister Albino rats were used. Right nephrectomy was performed in all groups. In the control group; I/R injury was not performed. In the I/R group; left renal pedicle was clamped for 45 minutes and then underwent 60 minutes and 24 hours of reperfusion. In the mannitol group; 1 mL 20% mannitol was given intravenously 15 minutes before clamping. In the udenafil group; 10-mg/kg udenafil was given orally 1 hour before clamping. Creatinine (Cr), blood urea nitrogen (BUN), Cr clearance, malondialdehyde, neutrophil gelatinase associated lipocalin (NGAL), histological examination and DNA damage (Comet Assay method) levels were compared in tissue, serum and urine samples. Udenafil had a better protective effect than mannitol according to biochemical parameters (Cr, BUN, Cr clearance, and NGAL levels) and histopathological findings when compared with the I/R group. In the Comet sampling analysis no significant difference was detected. Udenafil has a better renoprotective effect than mannitol against I/R injury and this effect supports more functional improvements. Further clinical trials are needed to demonstrate those effects and clinical utility of udenafil for that purpose in humans.

pubmed23n0910_927

Allospecific Memory B Cell Responses Are Dependent on Autophagy.

Long-lived, donor-reactive memory B cells (Bmems) can produce alloantibodies that mediate transplant injury. Autophagy, an intrinsic mechanism of cell organelle/component recycling, is required for Bmem survival in infectious and model antigen systems, but whether autophagy affects alloreactive Bmem is unknown. We studied mice with an inducible yellow fluorescent protein (YFP) reporter expressed under the activation-induced cytidine deaminase (AID) promoter active in B cells undergoing germinal center reactions. Up to 12 months after allogeneic sensitization, splenic YFP<sup+</sup B cells were predominantly IgD<sup-</sup IgM<sup-</sup IgG<sup+</sup and expressed CD73, CD80, and PD-L2, consistent with Bmems. Labeled cells contained significantly more cells with autophagosomes and more autophagosomes per cell than unlabeled, naïve B cells. To test for a functional link, we quantified alloantibody formation in mice with B cells conditionally deficient in the requisite autophagy gene ATG7. These experiments revealed absent B cell ATG7 (1) prevented B cell autophagy, (2) inhibited secondary alloantibody responses without altering primary alloantibody formation, and (3) diminished frequencies of alloreactive Bmems. Pharmacological autophagy inhibition with 3-methyladenine had similar effects on wild-type mice. Together with new documentation of increased autophagosomes within human Bmems, our data indicate that targeting autophagy has potential for eliminating donor-reactive Bmems in transplant recipients.

Allospecific Memory B Cell Responses Are Dependent on Autophagy. Long-lived, donor-reactive memory B cells (Bmems) can produce alloantibodies that mediate transplant injury. Autophagy, an intrinsic mechanism of cell organelle/component recycling, is required for Bmem survival in infectious and model antigen systems, but whether autophagy affects alloreactive Bmem is unknown. We studied mice with an inducible yellow fluorescent protein (YFP) reporter expressed under the activation-induced cytidine deaminase (AID) promoter active in B cells undergoing germinal center reactions. Up to 12 months after allogeneic sensitization, splenic YFP<sup+</sup B cells were predominantly IgD<sup-</sup IgM<sup-</sup IgG<sup+</sup and expressed CD73, CD80, and PD-L2, consistent with Bmems. Labeled cells contained significantly more cells with autophagosomes and more autophagosomes per cell than unlabeled, naïve B cells. To test for a functional link, we quantified alloantibody formation in mice with B cells conditionally deficient in the requisite autophagy gene ATG7. These experiments revealed absent B cell ATG7 (1) prevented B cell autophagy, (2) inhibited secondary alloantibody responses without altering primary alloantibody formation, and (3) diminished frequencies of alloreactive Bmems. Pharmacological autophagy inhibition with 3-methyladenine had similar effects on wild-type mice. Together with new documentation of increased autophagosomes within human Bmems, our data indicate that targeting autophagy has potential for eliminating donor-reactive Bmems in transplant recipients.

pubmed23n0029_26

Sphenoethmoidal encephalocele: a case report with review of the literature.

A rare case of sphenoethmoidal encephalocele with detailed pathological studies is reported. Only a few cases have been documented and were reviewed. The etiology and embryology of the congenital malformation remains speculative with an agreement on their development at an early embryonic stage. The mass presenting in the nasopharyngeal cavity remains often undetected unless causing respiratory embarrassment and is frequently misdiagnosed. Thorough radiological investigations detect the bone defect and outline the extent of the herniated mass and associated cerebral abnormalities. Small lesions have been successfully repaired. Larger ones prove less amenable to surgery and carry a high mortality.

Sphenoethmoidal encephalocele: a case report with review of the literature. A rare case of sphenoethmoidal encephalocele with detailed pathological studies is reported. Only a few cases have been documented and were reviewed. The etiology and embryology of the congenital malformation remains speculative with an agreement on their development at an early embryonic stage. The mass presenting in the nasopharyngeal cavity remains often undetected unless causing respiratory embarrassment and is frequently misdiagnosed. Thorough radiological investigations detect the bone defect and outline the extent of the herniated mass and associated cerebral abnormalities. Small lesions have been successfully repaired. Larger ones prove less amenable to surgery and carry a high mortality.

pubmed23n0699_24511

Nasal polyps do not contain olfactory structures.

Nasal polyposis can lead to olfactory dysfunction, either due to physical obstruction of the olfactory cleft or physiological disruption of the olfactory neuroepithelium. Where medical therapy has failed to relieve symptoms of nasal polyposis, surgical excision can be considered. However, removal of polyps medial to the middle turbinate is controversial: some believe this will relieve physical obstruction to odourants, others state that removal here risks excising olfactory neuroepithelium. We stained 25 nasal polypectomy samples from the area medial to the middle turbinate with olfactory marker protein. We confirmed that our staining method worked on normal olfactory tissue. However, no positive staining of nasal polyps was demonstrated. We conclude that nasal polyps medial to the middle turbinate do not contain olfactory neurons, and surgical excision is not contraindicated.

Nasal polyps do not contain olfactory structures. Nasal polyposis can lead to olfactory dysfunction, either due to physical obstruction of the olfactory cleft or physiological disruption of the olfactory neuroepithelium. Where medical therapy has failed to relieve symptoms of nasal polyposis, surgical excision can be considered. However, removal of polyps medial to the middle turbinate is controversial: some believe this will relieve physical obstruction to odourants, others state that removal here risks excising olfactory neuroepithelium. We stained 25 nasal polypectomy samples from the area medial to the middle turbinate with olfactory marker protein. We confirmed that our staining method worked on normal olfactory tissue. However, no positive staining of nasal polyps was demonstrated. We conclude that nasal polyps medial to the middle turbinate do not contain olfactory neurons, and surgical excision is not contraindicated.

pubmed23n0405_11692

Repair of deaminated bases in DNA.

Deamination of DNA bases can occur spontaneously, generating highly mutagenic lesions such as uracil, hypoxanthine, and xanthine. When cells are under oxidative stress that is induced either by oxidizing agents or by mitochondrial dysfunction, additional deamination products such as 5-hydroxymethyluracil (5-HMU) and 5-hydroxyuracil (5-OH-Ura) are formed. The cellular level of these highly mutagenic lesions is increased substantially when cells are exposed to DNA damaging agent, such as ionizing radiation, redox reagents, nitric oxide, and others. The cellular repair of deamination products is predominantly through the base excision repair (BER) pathway, a major cellular repair pathway that is initiated by lesion specific DNA glycosylases. In BER, the lesions are removed by the combined action of a DNA glycosylase and an AP endonuclease, leaving behind a one-base gap. The gapped product is then further repaired by the sequential action of DNA polymerase and DNA ligase. DNA glycosylases that recognize uracil, 5-OH-Ura, 5-HMU (derived from 5-methylcytosine) and a T/G mismatch (derived from a 5-methylcytosine/G pair) are present in most cells. Many of these glycosylases have been cloned and well characterized. In yeast and mammalian cells, hypoxanthine is efficiently removed by methylpurine N-glycosylase, and it is thought that BER might be an important pathway for the repair of hypoxanthine. In contrast, no glycosylase that can recognize xanthine has been identified in either yeast or mammalian cells. In Escherichia coli, the major enzyme activity that initiates the repair of hypoxanthine and xanthine is endonuclease V. Endonuclease V is an endonuclease that hydrolyzes the second phosphodiester bond 3' to the lesion. It is hypothesized that the cleaved DNA is further repaired through an alternative excision repair (AER) pathway that requires the participation of either a 5' endonuclease or a 3'-5' exonuclease to remove the damaged base. The repair process is then completed by the sequential actions of DNA polymerase and DNA ligase. Endonuclease V sequence homologs are present in all kingdoms, and it is conceivable that endonuclease V might also be a major enzyme that initiates the repair of hypoxanthine and xanthine in mammalian cells.

Repair of deaminated bases in DNA. Deamination of DNA bases can occur spontaneously, generating highly mutagenic lesions such as uracil, hypoxanthine, and xanthine. When cells are under oxidative stress that is induced either by oxidizing agents or by mitochondrial dysfunction, additional deamination products such as 5-hydroxymethyluracil (5-HMU) and 5-hydroxyuracil (5-OH-Ura) are formed. The cellular level of these highly mutagenic lesions is increased substantially when cells are exposed to DNA damaging agent, such as ionizing radiation, redox reagents, nitric oxide, and others. The cellular repair of deamination products is predominantly through the base excision repair (BER) pathway, a major cellular repair pathway that is initiated by lesion specific DNA glycosylases. In BER, the lesions are removed by the combined action of a DNA glycosylase and an AP endonuclease, leaving behind a one-base gap. The gapped product is then further repaired by the sequential action of DNA polymerase and DNA ligase. DNA glycosylases that recognize uracil, 5-OH-Ura, 5-HMU (derived from 5-methylcytosine) and a T/G mismatch (derived from a 5-methylcytosine/G pair) are present in most cells. Many of these glycosylases have been cloned and well characterized. In yeast and mammalian cells, hypoxanthine is efficiently removed by methylpurine N-glycosylase, and it is thought that BER might be an important pathway for the repair of hypoxanthine. In contrast, no glycosylase that can recognize xanthine has been identified in either yeast or mammalian cells. In Escherichia coli, the major enzyme activity that initiates the repair of hypoxanthine and xanthine is endonuclease V. Endonuclease V is an endonuclease that hydrolyzes the second phosphodiester bond 3' to the lesion. It is hypothesized that the cleaved DNA is further repaired through an alternative excision repair (AER) pathway that requires the participation of either a 5' endonuclease or a 3'-5' exonuclease to remove the damaged base. The repair process is then completed by the sequential actions of DNA polymerase and DNA ligase. Endonuclease V sequence homologs are present in all kingdoms, and it is conceivable that endonuclease V might also be a major enzyme that initiates the repair of hypoxanthine and xanthine in mammalian cells.

pubmed23n0672_3793

[Septate uteri: must we treat all of them?].

Septate uterus is the most common female genital malformation. It can be responsible for recurrent miscarriages and infertility. Hysteroscopic septoplasty is an effective procedure. It can prevent abortions and probably increase fertility. The hysteroscopic section of the septum has become a simple and safe technique, with the use of small hysteroscopes and that of bipolar energy with trained surgeons. These are the reasons why hysteroscopic septoplasty can be proposed to women with septate uterus and a desire for pregnancy.

[Septate uteri: must we treat all of them?]. Septate uterus is the most common female genital malformation. It can be responsible for recurrent miscarriages and infertility. Hysteroscopic septoplasty is an effective procedure. It can prevent abortions and probably increase fertility. The hysteroscopic section of the septum has become a simple and safe technique, with the use of small hysteroscopes and that of bipolar energy with trained surgeons. These are the reasons why hysteroscopic septoplasty can be proposed to women with septate uterus and a desire for pregnancy.

pubmed23n0866_14308

Combined gadoxetic acid and gadofosveset enhanced liver MRI for detection and characterization of liver metastases.

To compare gadoxetic acid alone and combined gadoxetic acid/gadofosveset trisodium-enhanced liver MRI for detection of metastases and differentiation of metastases from haemangiomas. Ninety-one patients underwent gadoxetic acid-enhanced liver MRI before and after additional injection of gadofosveset. First, two readers retrospectively identified metastases on gadoxetic acid alone enhanced delayed hepatobiliary phase T1-weighted images together with all other MR images (dynamic images, T2-weighted images, diffusion-weighted images). Second, readers assessed additional T1-weighted images obtained after administration of gadofosveset trisodium. For both interpretations, readers rated lesion conspicuity and confidence in differentiating metastases from haemangiomas. Results were compared using alternative free-response receiver-operating characteristic (AFROC) and conventional ROC methods. Histology and follow-up served as reference standard. There were 145 metastases and 16 haemangiomas. Both readers detected more metastases using combined gadoxetic acid/gadofosveset (reader 1 = 130; reader 2 = 124) compared to gadoxetic acid alone (reader 1 = 104; reader 2 = 103). Sensitivity of combined gadoxetic acid/gadofosveset (reader 1 = 90 %; reader 2 = 86 %) was higher than that of gadoxetic acid alone (reader 1 = 72 %; reader 2 = 71 %, both P &lt; 0.01). AFROC-AUC was higher for the combined technique (0.92 vs. 0.86, P &lt; 0.001). Sensitivity for correct differentiation of metastases from haemangiomas was higher for the combined technique (reader 1 = 98 %; reader 2 = 99 % vs. reader 1 = 86 %; reader 2 = 91 %, both P &lt; 0.01). ROC-AUC was significantly higher for the combined technique (reader 1 = 1.00; reader 2 = 1.00 vs. reader 1 = 0.87; reader 2 = 0.92, both P &lt; 0.01). Combined gadoxetic acid/gadofosveset-enhanced MRI improves detection and characterization of liver metastases compared to gadoxetic acid alone. • Combined gadoxetic acid and gadofosveset-enhanced liver MRI significantly improves detection of metastases. • The combined enhancement technique improves the accuracy to differentiate metastases from haemangiomas. • Prospective studies need to determine the clinical impact of the combined technique.

Combined gadoxetic acid and gadofosveset enhanced liver MRI for detection and characterization of liver metastases. To compare gadoxetic acid alone and combined gadoxetic acid/gadofosveset trisodium-enhanced liver MRI for detection of metastases and differentiation of metastases from haemangiomas. Ninety-one patients underwent gadoxetic acid-enhanced liver MRI before and after additional injection of gadofosveset. First, two readers retrospectively identified metastases on gadoxetic acid alone enhanced delayed hepatobiliary phase T1-weighted images together with all other MR images (dynamic images, T2-weighted images, diffusion-weighted images). Second, readers assessed additional T1-weighted images obtained after administration of gadofosveset trisodium. For both interpretations, readers rated lesion conspicuity and confidence in differentiating metastases from haemangiomas. Results were compared using alternative free-response receiver-operating characteristic (AFROC) and conventional ROC methods. Histology and follow-up served as reference standard. There were 145 metastases and 16 haemangiomas. Both readers detected more metastases using combined gadoxetic acid/gadofosveset (reader 1 = 130; reader 2 = 124) compared to gadoxetic acid alone (reader 1 = 104; reader 2 = 103). Sensitivity of combined gadoxetic acid/gadofosveset (reader 1 = 90 %; reader 2 = 86 %) was higher than that of gadoxetic acid alone (reader 1 = 72 %; reader 2 = 71 %, both P &lt; 0.01). AFROC-AUC was higher for the combined technique (0.92 vs. 0.86, P &lt; 0.001). Sensitivity for correct differentiation of metastases from haemangiomas was higher for the combined technique (reader 1 = 98 %; reader 2 = 99 % vs. reader 1 = 86 %; reader 2 = 91 %, both P &lt; 0.01). ROC-AUC was significantly higher for the combined technique (reader 1 = 1.00; reader 2 = 1.00 vs. reader 1 = 0.87; reader 2 = 0.92, both P &lt; 0.01). Combined gadoxetic acid/gadofosveset-enhanced MRI improves detection and characterization of liver metastases compared to gadoxetic acid alone. • Combined gadoxetic acid and gadofosveset-enhanced liver MRI significantly improves detection of metastases. • The combined enhancement technique improves the accuracy to differentiate metastases from haemangiomas. • Prospective studies need to determine the clinical impact of the combined technique.

pubmed23n0375_4740

[Nurturing young surgeons in a competitive society].

Creating and maintaining harmony between natural science and social science is very important for the establishment of a truly rich society in the 21st century. Young surgeons must acquire deep knowledge and excellent techniques, in addition to contributing to social development and human happiness. This paper outlines my personal views of what young surgeons need to do to succeed at all levels in the current competitive society.

[Nurturing young surgeons in a competitive society]. Creating and maintaining harmony between natural science and social science is very important for the establishment of a truly rich society in the 21st century. Young surgeons must acquire deep knowledge and excellent techniques, in addition to contributing to social development and human happiness. This paper outlines my personal views of what young surgeons need to do to succeed at all levels in the current competitive society.

pubmed23n0829_5079

Home Versus Away Competition: Effect on Psychophysiological Variables in Elite Rugby Union.

This study evaluated the effect of game venue and starting status on precompetitive psychophysiological measures in elite rugby union. Saliva samples were taken from players (starting XV, n = 15, and nonstarters, n = 9) on a control day and 90 min before 4 games played consecutively at home and away venues against local rivals and league leaders. Precompetition psychological states were assessed using the Competitive State Anxiety Inventory-2. The squad recorded 2 wins (home) and 2 losses (away) over the study period. Calculated effect sizes (ESs) showed higher pregame cortisol- (C) and testosterone- (T) difference values before all games than on a baseline control day (ES 0.7-1.5). Similar findings were observed for cognitive and somatic anxiety. Small between-venues C differences were observed in starting XV players (ES 0.2-0.25). Conversely, lower home T- (ES 0.95) and higher away C- (ES 0.6) difference values were observed in nonstarters. Lower T-difference values were apparent in nonstarters (vs starting XV) before home games, providing evidence of a between-groups effect (ES 0.92). Findings show an anticipatory rise in psychophysiological variables before competition. Knowledge of starting status appears a moderating factor in the magnitude of player endocrine response between home and away games.

Home Versus Away Competition: Effect on Psychophysiological Variables in Elite Rugby Union. This study evaluated the effect of game venue and starting status on precompetitive psychophysiological measures in elite rugby union. Saliva samples were taken from players (starting XV, n = 15, and nonstarters, n = 9) on a control day and 90 min before 4 games played consecutively at home and away venues against local rivals and league leaders. Precompetition psychological states were assessed using the Competitive State Anxiety Inventory-2. The squad recorded 2 wins (home) and 2 losses (away) over the study period. Calculated effect sizes (ESs) showed higher pregame cortisol- (C) and testosterone- (T) difference values before all games than on a baseline control day (ES 0.7-1.5). Similar findings were observed for cognitive and somatic anxiety. Small between-venues C differences were observed in starting XV players (ES 0.2-0.25). Conversely, lower home T- (ES 0.95) and higher away C- (ES 0.6) difference values were observed in nonstarters. Lower T-difference values were apparent in nonstarters (vs starting XV) before home games, providing evidence of a between-groups effect (ES 0.92). Findings show an anticipatory rise in psychophysiological variables before competition. Knowledge of starting status appears a moderating factor in the magnitude of player endocrine response between home and away games.

pubmed23n0220_17300

Vasoactivity generated from platelets by immune complexes in pre-eclampsia.

Circulating immune complexes in excess of the equivalent of 20 micrograms/ml heat-aggregated IgG were found in fourteen out of twenty patients diagnosed as having preeclampsia. Only six of the nineteen controls tested had similar levels of immune complexes. Recent studies have established that concentrations of heat aggregated IgG in excess of 20 micrograms/ml activate human platelets to release sufficient concentrations of vasoactive agents to constrict a human blood vessel in vitro. It is therefore suggested that in vivo platelet activation by circulating immune complexes may release sufficient concentrations of vasoactive agents to contribute to the hypertension in pre-eclampsia.

Vasoactivity generated from platelets by immune complexes in pre-eclampsia. Circulating immune complexes in excess of the equivalent of 20 micrograms/ml heat-aggregated IgG were found in fourteen out of twenty patients diagnosed as having preeclampsia. Only six of the nineteen controls tested had similar levels of immune complexes. Recent studies have established that concentrations of heat aggregated IgG in excess of 20 micrograms/ml activate human platelets to release sufficient concentrations of vasoactive agents to constrict a human blood vessel in vitro. It is therefore suggested that in vivo platelet activation by circulating immune complexes may release sufficient concentrations of vasoactive agents to contribute to the hypertension in pre-eclampsia.

pubmed23n0320_19246

Mapping of vascular dendritic cells in atherosclerotic arteries suggests their involvement in local immune-inflammatory reactions.

We previously demonstrated that vascular dendritic cells (VDCs) are present in the intima of large arteries and that their numbers are increased in atherosclerotic lesions. This study was undertaken to determine whether VDCs are involved in immune-mediated reactions in atherogenesis. Specimens of carotid artery and aorta were obtained at operation. VDCs were identified with anti-CD1a or with S-100. Co-localisation of VDCs with different intimal cells, including T-cells and macrophages, was studied using a double immunostaining procedure. In areas where the co-localising cells were detected, the peculiarities of expression of HLA-DR, ICAM-1, VCAM-1 were examined. In all the atherosclerotic plaques, VDCs were seen in contact with T-cells, but these co-localising cells were irregularly distributed and were mainly found in zones of neovascularisation containing inflammatory infiltrates. In other areas, T-cell/VDC co-localisation was rarely detected but VDCs were often found in contact with macrophages. VDCs were detected also in the media beneath atherosclerotic lesions and in the adventitia, where they were mostly around vasa vasorum, especially in areas exhibiting signs of acute inflammation. In these areas VDCs expressed ICAM-1, VCAM-1 and were in contact with T-cells. In both plaques and in the adventitia, the areas with co-localising VDCs and T-cells corresponded to the areas with HLA-DR expression. The results suggest that VDCs are involved in T-cell activation in atherogenesis. There are two regions within the arterial wall where VDC/T-cell co-localisation mostly occurs, namely, in zones of neovascularisation containing inflammatory infiltrates located within atherosclerotic lesions, and in areas with inflammatory infiltrates around vasa vasorum in the adventitia. Possibly, some intimal VDCs migrate through the media and adventitia to adjacent lymph nodes where they present atherosclerosis associated antigens. We also speculate that VDC/macrophage contacts are essential in processing immune information in atherogenesis.

Mapping of vascular dendritic cells in atherosclerotic arteries suggests their involvement in local immune-inflammatory reactions. We previously demonstrated that vascular dendritic cells (VDCs) are present in the intima of large arteries and that their numbers are increased in atherosclerotic lesions. This study was undertaken to determine whether VDCs are involved in immune-mediated reactions in atherogenesis. Specimens of carotid artery and aorta were obtained at operation. VDCs were identified with anti-CD1a or with S-100. Co-localisation of VDCs with different intimal cells, including T-cells and macrophages, was studied using a double immunostaining procedure. In areas where the co-localising cells were detected, the peculiarities of expression of HLA-DR, ICAM-1, VCAM-1 were examined. In all the atherosclerotic plaques, VDCs were seen in contact with T-cells, but these co-localising cells were irregularly distributed and were mainly found in zones of neovascularisation containing inflammatory infiltrates. In other areas, T-cell/VDC co-localisation was rarely detected but VDCs were often found in contact with macrophages. VDCs were detected also in the media beneath atherosclerotic lesions and in the adventitia, where they were mostly around vasa vasorum, especially in areas exhibiting signs of acute inflammation. In these areas VDCs expressed ICAM-1, VCAM-1 and were in contact with T-cells. In both plaques and in the adventitia, the areas with co-localising VDCs and T-cells corresponded to the areas with HLA-DR expression. The results suggest that VDCs are involved in T-cell activation in atherogenesis. There are two regions within the arterial wall where VDC/T-cell co-localisation mostly occurs, namely, in zones of neovascularisation containing inflammatory infiltrates located within atherosclerotic lesions, and in areas with inflammatory infiltrates around vasa vasorum in the adventitia. Possibly, some intimal VDCs migrate through the media and adventitia to adjacent lymph nodes where they present atherosclerosis associated antigens. We also speculate that VDC/macrophage contacts are essential in processing immune information in atherogenesis.

pubmed23n0722_2277

A novel mammalian social structure in Indo-Pacific bottlenose dolphins (Tursiops sp.): complex male alliances in an open social network.

Terrestrial mammals with differentiated social relationships live in 'semi-closed groups' that occasionally accept new members emigrating from other groups. Bottlenose dolphins (Tursiops sp.) in Shark Bay, Western Australia, exhibit a fission-fusion grouping pattern with strongly differentiated relationships, including nested male alliances. Previous studies failed to detect a group membership 'boundary', suggesting that the dolphins live in an open social network. However, two alternative hypotheses have not been excluded. The community defence model posits that the dolphins live in a large semi-closed 'chimpanzee-like' community defended by males and predicts that a dominant alliance(s) will range over the entire community range. The mating season defence model predicts that alliances will defend mating-season territories or sets of females. Here, both models are tested and rejected: no alliances ranged over the entire community range and alliances showed extensive overlap in mating season ranges and consorted females. The Shark Bay dolphins, therefore, present a combination of traits that is unique among mammals: complex male alliances in an open social network. The open social network of dolphins is linked to their relatively low costs of locomotion. This reveals a surprising and previously unrecognized convergence between adaptations reducing travel costs and complex intergroup-alliance relationships in dolphins, elephants and humans.

A novel mammalian social structure in Indo-Pacific bottlenose dolphins (Tursiops sp.): complex male alliances in an open social network. Terrestrial mammals with differentiated social relationships live in 'semi-closed groups' that occasionally accept new members emigrating from other groups. Bottlenose dolphins (Tursiops sp.) in Shark Bay, Western Australia, exhibit a fission-fusion grouping pattern with strongly differentiated relationships, including nested male alliances. Previous studies failed to detect a group membership 'boundary', suggesting that the dolphins live in an open social network. However, two alternative hypotheses have not been excluded. The community defence model posits that the dolphins live in a large semi-closed 'chimpanzee-like' community defended by males and predicts that a dominant alliance(s) will range over the entire community range. The mating season defence model predicts that alliances will defend mating-season territories or sets of females. Here, both models are tested and rejected: no alliances ranged over the entire community range and alliances showed extensive overlap in mating season ranges and consorted females. The Shark Bay dolphins, therefore, present a combination of traits that is unique among mammals: complex male alliances in an open social network. The open social network of dolphins is linked to their relatively low costs of locomotion. This reveals a surprising and previously unrecognized convergence between adaptations reducing travel costs and complex intergroup-alliance relationships in dolphins, elephants and humans.

pubmed23n0673_5713

Epileptic seizures: Quakes of the brain?

A dynamical analogy supported by five scale-free statistics (the Gutenberg-Richter distribution of event sizes, the distribution of interevent intervals, the Omori and inverse Omori laws, and the conditional waiting time until the next event) is shown to exist between two classes of seizures ("focal" in humans and generalized in animals) and earthquakes. Increments in excitatory interneuronal coupling in animals expose the system's dependence on this parameter and its dynamical transmutability: moderate increases lead to power-law behavior of seizure energy and interevent times, while marked ones to scale-free (power-law) coextensive with characteristic scales and events. The coextensivity of power law and characteristic size regimes is predicted by models of coupled heterogeneous threshold oscillators of relaxation and underscores the role of coupling strength in shaping the dynamics of these systems.

Epileptic seizures: Quakes of the brain? A dynamical analogy supported by five scale-free statistics (the Gutenberg-Richter distribution of event sizes, the distribution of interevent intervals, the Omori and inverse Omori laws, and the conditional waiting time until the next event) is shown to exist between two classes of seizures ("focal" in humans and generalized in animals) and earthquakes. Increments in excitatory interneuronal coupling in animals expose the system's dependence on this parameter and its dynamical transmutability: moderate increases lead to power-law behavior of seizure energy and interevent times, while marked ones to scale-free (power-law) coextensive with characteristic scales and events. The coextensivity of power law and characteristic size regimes is predicted by models of coupled heterogeneous threshold oscillators of relaxation and underscores the role of coupling strength in shaping the dynamics of these systems.

pubmed23n0352_21190

In vitro and in vivo extrapolations of genotoxin exposures: consideration of factors which influence dose-response thresholds.

The concept of a threshold of activity of a genotoxic agent is primarily based upon considerations of protective mechanisms and multiple cellular targets, which require inactivation before a toxic response is produced. In this paper, we have considered and evaluated the influences of compound metabolism, DNA lesion formation, mutation induction and sequence content, aneuploidy induction and the influence of repair enzymes upon genetic endpoints produced by both DNA reactive chemicals and by those chemicals which modify non-DNA cellular targets. Thresholds of activity have been evaluated by critical analysis of the published literature and original data analysing both the role of sequence context upon point mutation induction and DNA repair mechanisms upon the sensitivity of cultured cells to the induction of aneuploidy. In the case of DNA reactive chemicals, the presence of a threshold of chemical activity will be dependent upon cellular activities such as those of the Phase II enzymes reducing the activity of chemicals before lesion formation takes place and/or those of the DNA repair enzymes which reduce the proportion of DNA lesions which are processed into DNA sequence changes. Under such conditions, a given exposure of a DNA reactive chemical does not produce a linear or semi-linear increase in DNA lesions or in mutation frequency. However, even when these protective mechanisms are overwhelmed by the high exposures of genotoxic chemicals the biological effects of a genotoxin may be influenced by the sequence context of the gene under consideration. Here, we demonstrate that point mutations are detected at relatively higher frequencies in the non-coding introns compared with the coding exons. Many of the base changes detected in the exons do not produce amino acid changes in the proteins coded for by the genes being monitored for mutation induction. Both sequence context and the types of base changes induced may provide a "buffering" effect reducing the biological consequences of mutation induction. Spindle damaging chemicals, such as colcemid and vinblastine, induce aneuploidy by modifying the numbers of spindle fibres which regulate the segregation of chromosomes during mitosis and meiosis. The redundancy of spindle fibres in the dividing mammalian cell leads to the prediction that only chemical exposures which damage most, if not all, of the fibres will lead to the induction of polyploidy and/or aneuploidy. Such predicted thresholds of chemical activity can be observed when both chromosome loss and non-disjunction are measured in wild type cultures. However, we observed a substantial increase in sensitivity to aneugenic chemicals when measurements were made in primary cell cultures derived from xerodoma pigmentosum and trichothiodystrophy patients. Further studies are necessary to evaluate the consequences of the genetic background of tester strains upon the nature of the dose-response curve of aneugenic chemicals.

In vitro and in vivo extrapolations of genotoxin exposures: consideration of factors which influence dose-response thresholds. The concept of a threshold of activity of a genotoxic agent is primarily based upon considerations of protective mechanisms and multiple cellular targets, which require inactivation before a toxic response is produced. In this paper, we have considered and evaluated the influences of compound metabolism, DNA lesion formation, mutation induction and sequence content, aneuploidy induction and the influence of repair enzymes upon genetic endpoints produced by both DNA reactive chemicals and by those chemicals which modify non-DNA cellular targets. Thresholds of activity have been evaluated by critical analysis of the published literature and original data analysing both the role of sequence context upon point mutation induction and DNA repair mechanisms upon the sensitivity of cultured cells to the induction of aneuploidy. In the case of DNA reactive chemicals, the presence of a threshold of chemical activity will be dependent upon cellular activities such as those of the Phase II enzymes reducing the activity of chemicals before lesion formation takes place and/or those of the DNA repair enzymes which reduce the proportion of DNA lesions which are processed into DNA sequence changes. Under such conditions, a given exposure of a DNA reactive chemical does not produce a linear or semi-linear increase in DNA lesions or in mutation frequency. However, even when these protective mechanisms are overwhelmed by the high exposures of genotoxic chemicals the biological effects of a genotoxin may be influenced by the sequence context of the gene under consideration. Here, we demonstrate that point mutations are detected at relatively higher frequencies in the non-coding introns compared with the coding exons. Many of the base changes detected in the exons do not produce amino acid changes in the proteins coded for by the genes being monitored for mutation induction. Both sequence context and the types of base changes induced may provide a "buffering" effect reducing the biological consequences of mutation induction. Spindle damaging chemicals, such as colcemid and vinblastine, induce aneuploidy by modifying the numbers of spindle fibres which regulate the segregation of chromosomes during mitosis and meiosis. The redundancy of spindle fibres in the dividing mammalian cell leads to the prediction that only chemical exposures which damage most, if not all, of the fibres will lead to the induction of polyploidy and/or aneuploidy. Such predicted thresholds of chemical activity can be observed when both chromosome loss and non-disjunction are measured in wild type cultures. However, we observed a substantial increase in sensitivity to aneugenic chemicals when measurements were made in primary cell cultures derived from xerodoma pigmentosum and trichothiodystrophy patients. Further studies are necessary to evaluate the consequences of the genetic background of tester strains upon the nature of the dose-response curve of aneugenic chemicals.

pubmed23n0902_22516

[Detection and prevention of blindness in patients with glaucoma is a socio-economical challenge].

Glaucoma is the most common cause of incurable blindness. If untreated, glaucoma leads to permanent vision loss starting with unnoticeable blind spots in the peripheral visual field. Due to the age-related nature of glaucoma, the disease is becoming an increasing socio-economical challenge. Extrapolations have predicted more than 110 million people with glaucoma in 2040. Twice as many patients were diagnosed with glaucoma in Denmark in 2011 as in 1996. Light is shedded on the need for improved awareness of the significant present and future burden of glaucoma.

[Detection and prevention of blindness in patients with glaucoma is a socio-economical challenge]. Glaucoma is the most common cause of incurable blindness. If untreated, glaucoma leads to permanent vision loss starting with unnoticeable blind spots in the peripheral visual field. Due to the age-related nature of glaucoma, the disease is becoming an increasing socio-economical challenge. Extrapolations have predicted more than 110 million people with glaucoma in 2040. Twice as many patients were diagnosed with glaucoma in Denmark in 2011 as in 1996. Light is shedded on the need for improved awareness of the significant present and future burden of glaucoma.

pubmed23n0256_5007

Assessment of lymphokine profiles in activated lymphocytes by semiquantitative PCR.

Several methods are currently used to detect the expression of specific mRNAs in leukocytes. While Northern blot analysis and RNase protection assays are commonly chosen for quantitative assessment of mRNA levels, these methods require a significant quantity of RNA, making their use unfeasible when limiting numbers of cells are available. Alternatively, use of the reverse transcription-polymerase chain reaction (RT-PCR) technique allows detection of specific mRNAs even at low copy number. It is, however, difficult to establish the conditions which allow consistent semi-quantitative assessment of specific mRNA expression, using the RT-PCR method. We report here a modification of the RT-PCR technique, which has enabled us to compare lymphokine mRNA expression profiles in mixed cell populations activated either in vivo or in vitro. This modification is based on the use of standard RNAs generated by in vitro transcription of size-modified CD3 and IFN-gamma-specific PCR products subcloned into the pGEM3 plasmid. Equal amounts of standard RNAs are introduced into each sample, reverse transcribed and co-amplified with cellular mRNA to control the reproducibility and efficiency of the method. The template therefore follows the cellular RNA through all steps of the analysis, and the corresponding 32P-labelled PCR products are subsequently separated by PAGE procedure. The amount of radioactivity incorporated into lymphokine-specific bands is determined by densitometry and normalized against the density of standard bands. Under optimal PCR conditions this method is linear over a 50-fold range of dilutions. The technique is specific, reproducible and fast, allowing an analysis of lymphokine-specific mRNA profiles in samples containing 10(4)-10(6) cells.

Assessment of lymphokine profiles in activated lymphocytes by semiquantitative PCR. Several methods are currently used to detect the expression of specific mRNAs in leukocytes. While Northern blot analysis and RNase protection assays are commonly chosen for quantitative assessment of mRNA levels, these methods require a significant quantity of RNA, making their use unfeasible when limiting numbers of cells are available. Alternatively, use of the reverse transcription-polymerase chain reaction (RT-PCR) technique allows detection of specific mRNAs even at low copy number. It is, however, difficult to establish the conditions which allow consistent semi-quantitative assessment of specific mRNA expression, using the RT-PCR method. We report here a modification of the RT-PCR technique, which has enabled us to compare lymphokine mRNA expression profiles in mixed cell populations activated either in vivo or in vitro. This modification is based on the use of standard RNAs generated by in vitro transcription of size-modified CD3 and IFN-gamma-specific PCR products subcloned into the pGEM3 plasmid. Equal amounts of standard RNAs are introduced into each sample, reverse transcribed and co-amplified with cellular mRNA to control the reproducibility and efficiency of the method. The template therefore follows the cellular RNA through all steps of the analysis, and the corresponding 32P-labelled PCR products are subsequently separated by PAGE procedure. The amount of radioactivity incorporated into lymphokine-specific bands is determined by densitometry and normalized against the density of standard bands. Under optimal PCR conditions this method is linear over a 50-fold range of dilutions. The technique is specific, reproducible and fast, allowing an analysis of lymphokine-specific mRNA profiles in samples containing 10(4)-10(6) cells.

pubmed23n0254_14762

Mechanical and contractile properties of in situ localized mesenteric arteries in normotensive and spontaneously hypertensive rats.

An in situ model was developed for studying mechanical properties of mesenteric arteries in rats. A branch of the mesenteric artery was exposed and dissected in normotensive (WKY) and spontaneously hypertensive rats (SHR). A catheter was introduced into the larger branch of the mesenteric artery and connected to a pressure chamber. The artery was submitted to transmural pressures ranging from 0 to 200 mmHg per steps of 25 mmHg and observed using a microscope-video-camera system. The diameter-pressure relations were established under basal conditions, under contraction (phenylephrine 10(-6) M), and after abolition of the smooth muscle tone by potassium cyanide (KCN, 0.1 mg/mL). The arterial segment was then fixed (glutaraldehyde 2.5%), and the wall cross-sectional areas were measured in transverse sections. Compliances, distensibility, wall tensions, and wall stresses were calculated from diameter, pressure, and media thickness values under three conditions. Active tension and active stress were defined as differences in wall stresses and wall tensions calculated under passive and active conditions. Comparison of WKY and SHR when arteries were studied at the respective operating pressure indicates (1) thicker and stiffer mesenteric arteries in SHRs than in WKY rats, (2) similar wall stresses in mesenteric arteries from WKY and SHRs despite larger wall tensions in the hypertensive group, and (3) larger contractility to phenylephrine in SHRs than in WKY mesenteric arteries.

Mechanical and contractile properties of in situ localized mesenteric arteries in normotensive and spontaneously hypertensive rats. An in situ model was developed for studying mechanical properties of mesenteric arteries in rats. A branch of the mesenteric artery was exposed and dissected in normotensive (WKY) and spontaneously hypertensive rats (SHR). A catheter was introduced into the larger branch of the mesenteric artery and connected to a pressure chamber. The artery was submitted to transmural pressures ranging from 0 to 200 mmHg per steps of 25 mmHg and observed using a microscope-video-camera system. The diameter-pressure relations were established under basal conditions, under contraction (phenylephrine 10(-6) M), and after abolition of the smooth muscle tone by potassium cyanide (KCN, 0.1 mg/mL). The arterial segment was then fixed (glutaraldehyde 2.5%), and the wall cross-sectional areas were measured in transverse sections. Compliances, distensibility, wall tensions, and wall stresses were calculated from diameter, pressure, and media thickness values under three conditions. Active tension and active stress were defined as differences in wall stresses and wall tensions calculated under passive and active conditions. Comparison of WKY and SHR when arteries were studied at the respective operating pressure indicates (1) thicker and stiffer mesenteric arteries in SHRs than in WKY rats, (2) similar wall stresses in mesenteric arteries from WKY and SHRs despite larger wall tensions in the hypertensive group, and (3) larger contractility to phenylephrine in SHRs than in WKY mesenteric arteries.

pubmed23n0839_24812

Cultivating the uncultured: growing the recalcitrant cluster-2 Frankia strains.

The repeated failures reported in cultivating some microbial lineages are a major challenge in microbial ecology and probably linked, in the case of Frankia microsymbionts to atypical patterns of auxotrophy. Comparative genomics of the so far uncultured cluster-2 Candidatus Frankia datiscae Dg1, with cultivated Frankiae has revealed genome reduction, but no obvious physiological impairments. A direct physiological assay on nodule tissues from Coriaria myrtifolia infected with a closely-related strain permitted the identification of a requirement for alkaline conditions. A high pH growth medium permitted the recovery of a slow-growing actinobacterium. The strain obtained, called BMG5.1, has short hyphae, produced diazovesicles in nitrogen-free media, and fulfilled Koch's postulates by inducing effective nodules on axenically grown Coriaria spp. and Datisca glomerata. Analysis of the draft genome confirmed its close proximity to the Candidatus Frankia datiscae Dg1 genome with the absence of 38 genes (trehalose synthase, fumarylacetoacetase, etc) in BMG5.1 and the presence of 77 other genes (CRISPR, lanthionine synthase, glutathione synthetase, catalase, Na+/H+ antiporter, etc) not found in Dg1. A multi-gene phylogeny placed the two cluster-2 strains together at the root of the Frankia radiation.

Cultivating the uncultured: growing the recalcitrant cluster-2 Frankia strains. The repeated failures reported in cultivating some microbial lineages are a major challenge in microbial ecology and probably linked, in the case of Frankia microsymbionts to atypical patterns of auxotrophy. Comparative genomics of the so far uncultured cluster-2 Candidatus Frankia datiscae Dg1, with cultivated Frankiae has revealed genome reduction, but no obvious physiological impairments. A direct physiological assay on nodule tissues from Coriaria myrtifolia infected with a closely-related strain permitted the identification of a requirement for alkaline conditions. A high pH growth medium permitted the recovery of a slow-growing actinobacterium. The strain obtained, called BMG5.1, has short hyphae, produced diazovesicles in nitrogen-free media, and fulfilled Koch's postulates by inducing effective nodules on axenically grown Coriaria spp. and Datisca glomerata. Analysis of the draft genome confirmed its close proximity to the Candidatus Frankia datiscae Dg1 genome with the absence of 38 genes (trehalose synthase, fumarylacetoacetase, etc) in BMG5.1 and the presence of 77 other genes (CRISPR, lanthionine synthase, glutathione synthetase, catalase, Na+/H+ antiporter, etc) not found in Dg1. A multi-gene phylogeny placed the two cluster-2 strains together at the root of the Frankia radiation.

pubmed23n0678_2323

Whitnall ligament anatomy revisited.

To examine the concentrations of elastic fibres in Whitnall ligament and the intermuscular transverse ligament (ITL), and to examine the anatomic relationships between Whitnall ligament and its surrounding structures. Observational anatomic study. Full thickness sagittal sections of 12 upper eyelids (6 right and 6 left), fixed in 10 % buffered formalin, were studied in 10 Asian cadavers (mean age at death: 77.1 years, range: 62-92 years). We analysed the concentrations of elastic fibres in Whitnall ligament and the ITL, as well as examined the relationship between Whitnall ligament and the connective tissue septa, the ITL and the levator palpebrae superioris (LPS) muscle. Concentrations of elastic fibres in Whitnall ligament and the ITL, relationship between Whitnall ligament and its surrounding structures. The concentration of elastic fibres in Whitnall ligament was low compared to the ITL. In most specimens, both Whitnall ligament and the ITL were loosely attached to the LPS muscle through areas of adipose tissue or cavities. Although in all specimens, connective tissue septa were demonstrated next to Whitnall ligament, they were more often directed parallel to the ligament rather than radially to it. Both Whitnall ligament, with its low concentration of elastic fibres, and the ITL, with its high content of elastic fibres, show loose attachments to the LPS muscle through areas of adipose tissue or cavities. Whitnall ligament may work as the check ligament of the LPS muscle as well as part of the pulley system with the ITL.

Whitnall ligament anatomy revisited. To examine the concentrations of elastic fibres in Whitnall ligament and the intermuscular transverse ligament (ITL), and to examine the anatomic relationships between Whitnall ligament and its surrounding structures. Observational anatomic study. Full thickness sagittal sections of 12 upper eyelids (6 right and 6 left), fixed in 10 % buffered formalin, were studied in 10 Asian cadavers (mean age at death: 77.1 years, range: 62-92 years). We analysed the concentrations of elastic fibres in Whitnall ligament and the ITL, as well as examined the relationship between Whitnall ligament and the connective tissue septa, the ITL and the levator palpebrae superioris (LPS) muscle. Concentrations of elastic fibres in Whitnall ligament and the ITL, relationship between Whitnall ligament and its surrounding structures. The concentration of elastic fibres in Whitnall ligament was low compared to the ITL. In most specimens, both Whitnall ligament and the ITL were loosely attached to the LPS muscle through areas of adipose tissue or cavities. Although in all specimens, connective tissue septa were demonstrated next to Whitnall ligament, they were more often directed parallel to the ligament rather than radially to it. Both Whitnall ligament, with its low concentration of elastic fibres, and the ITL, with its high content of elastic fibres, show loose attachments to the LPS muscle through areas of adipose tissue or cavities. Whitnall ligament may work as the check ligament of the LPS muscle as well as part of the pulley system with the ITL.

pubmed23n0044_13624

[Effects of agonists and antagonists of benzodiazepine, GABA and NMDA receptors, on caffeine-induced seizures in mice].

In mice, tonic convulsive seizure induced by intravenous administration of caffeine (adenosine A1, A2 receptors antagonist) was significantly potentiated by any one of L-PIA (adenosine A1 receptor agonist), NECA (adenosine A2 receptor agonist) and 2-ClAd (adenosine A1, A2 receptors agonist). The caffeine-induced seizure was unaffected by diazepam (benzodiazepine receptor agonist), but was inhibited by Ro 15-1788 (antagonist or partial agonist). beta-DMCM (antagonist or inverse agonist) increased the seizure. Muscimol (GABA-a receptor agonist), baclofen (GABA-b receptor agonist) and AOAA (GABA transaminase inhibitor) did not show significant effect on caffeine-induced convulsion. Bicuculline (GABA-a receptor antagonist) and picrotoxin (chloride channel blocker) significantly potentiated the convulsion at the doses which did not induce it. Caffeine-induced convulsion was potentiated by NMDA with its non-convulsive dose. CPP (competitive NMDA receptor antagonist) and MK-801 (non-competitive NMDA receptor antagonist) significantly inhibited the seizures. These results suggest that caffeine-induced seizure is not caused by blockade of adenosine receptors. Caffeine may act to beta-carboline sensitive benzodiazepine receptor (Type 1) which has no linkage with GABA-a receptor. Furthermore, it is implied that caffeine plays some role at NMDA receptor calcium ion channel complex.

[Effects of agonists and antagonists of benzodiazepine, GABA and NMDA receptors, on caffeine-induced seizures in mice]. In mice, tonic convulsive seizure induced by intravenous administration of caffeine (adenosine A1, A2 receptors antagonist) was significantly potentiated by any one of L-PIA (adenosine A1 receptor agonist), NECA (adenosine A2 receptor agonist) and 2-ClAd (adenosine A1, A2 receptors agonist). The caffeine-induced seizure was unaffected by diazepam (benzodiazepine receptor agonist), but was inhibited by Ro 15-1788 (antagonist or partial agonist). beta-DMCM (antagonist or inverse agonist) increased the seizure. Muscimol (GABA-a receptor agonist), baclofen (GABA-b receptor agonist) and AOAA (GABA transaminase inhibitor) did not show significant effect on caffeine-induced convulsion. Bicuculline (GABA-a receptor antagonist) and picrotoxin (chloride channel blocker) significantly potentiated the convulsion at the doses which did not induce it. Caffeine-induced convulsion was potentiated by NMDA with its non-convulsive dose. CPP (competitive NMDA receptor antagonist) and MK-801 (non-competitive NMDA receptor antagonist) significantly inhibited the seizures. These results suggest that caffeine-induced seizure is not caused by blockade of adenosine receptors. Caffeine may act to beta-carboline sensitive benzodiazepine receptor (Type 1) which has no linkage with GABA-a receptor. Furthermore, it is implied that caffeine plays some role at NMDA receptor calcium ion channel complex.

pubmed23n0987_16211

Increasing Rates of Diagnosis, Substantial Co-Occurrence, and Variable Treatment Patterns of Eosinophilic Gastritis, Gastroenteritis, and Colitis Based on 10-Year Data Across a Multicenter Consortium.

The literature related to eosinophilic gastritis (EG), gastroenteritis (EGE), and colitis (EC) is limited. We aimed to characterize rates of diagnosis, clinical features, and initial treatments for patients with EG, EGE, and EC. In this retrospective study, data were collected from 6 centers in the Consortium of Eosinophilic Gastrointestinal Researchers from 2005 to 2016. We analyzed demographics, time trends in diagnosis, medical history, presenting symptoms, disease overlap, and initial treatment patterns/responses. Of 373 subjects (317 children and 56 adults), 38% had EG, 33% EGE, and 29% EC. Rates of diagnosis of all diseases increased over time. There was no male predominance, and the majority of subjects had atopy. Presenting symptoms were similar between diseases with nausea/vomiting and abdominal pain, the most common. One hundred fifty-four subjects (41%) had eosinophilic inflammation outside of their primary disease location with the esophagus the second most common gastrointestinal (GI) segment involved. Multisite inflammation was more common in children than in adults (68% vs 37%; P &lt; 0.001). Initial treatment patterns varied highly between centers. One hundred-nine subjects (29%) had follow-up within 6 months, and the majority had clinical, endoscopic, and histologic improvements. In this cohort, EG, EGE, and EC were diagnosed more frequently over time, and inflammation of GI segments outside the primary disease site co-occurrence of atopy was common with a lack of male predominance. Symptoms were similar between diseases, and initial treatment strategies were highly variable. Future investigation should assess the cause of the increased prevalence of eosinophilic GI disorders and prospectively assess outcomes to establish treatment algorithms.

Increasing Rates of Diagnosis, Substantial Co-Occurrence, and Variable Treatment Patterns of Eosinophilic Gastritis, Gastroenteritis, and Colitis Based on 10-Year Data Across a Multicenter Consortium. The literature related to eosinophilic gastritis (EG), gastroenteritis (EGE), and colitis (EC) is limited. We aimed to characterize rates of diagnosis, clinical features, and initial treatments for patients with EG, EGE, and EC. In this retrospective study, data were collected from 6 centers in the Consortium of Eosinophilic Gastrointestinal Researchers from 2005 to 2016. We analyzed demographics, time trends in diagnosis, medical history, presenting symptoms, disease overlap, and initial treatment patterns/responses. Of 373 subjects (317 children and 56 adults), 38% had EG, 33% EGE, and 29% EC. Rates of diagnosis of all diseases increased over time. There was no male predominance, and the majority of subjects had atopy. Presenting symptoms were similar between diseases with nausea/vomiting and abdominal pain, the most common. One hundred fifty-four subjects (41%) had eosinophilic inflammation outside of their primary disease location with the esophagus the second most common gastrointestinal (GI) segment involved. Multisite inflammation was more common in children than in adults (68% vs 37%; P &lt; 0.001). Initial treatment patterns varied highly between centers. One hundred-nine subjects (29%) had follow-up within 6 months, and the majority had clinical, endoscopic, and histologic improvements. In this cohort, EG, EGE, and EC were diagnosed more frequently over time, and inflammation of GI segments outside the primary disease site co-occurrence of atopy was common with a lack of male predominance. Symptoms were similar between diseases, and initial treatment strategies were highly variable. Future investigation should assess the cause of the increased prevalence of eosinophilic GI disorders and prospectively assess outcomes to establish treatment algorithms.

pubmed23n0790_17502

Automated ribotyping and antibiotic resistance determining of Bacillus spp from conjunctiva of diabetic patients.

We aimed to characterize the phenotype and genotype of Bacillus spp isolated from diabetic patients' eyes, by studying the drug sensitivity patterns with a disc-diffusion method. Fifty eyes of 25 patients with type II diabetes mellitus, with at least 10 years of diabetes history, were included in the study. We analyzed the eyes for the presence of Bacillus spp.; presumptive isolates were identified by morphological, and biochemical tests, and confirmed by the VITEK system. Automated EcoRI ribotyping was performed with a RiboPrinter(®) Microbial Characterization System. We determined the antibiotic resistance of the isolates by the Kirby-Bauer disc diffusion test. Seven out of 25 patients were on insulin treatment; 7 on oral anti-diabetic medication; and 11 on combination therapy of insulin and oral medications. Among the 28 Bacillus spp isolates, 14 were B. cereus, 11 were B. pumilus, 2 were B. mojavensis and 1 was B. subtilis. Almost all the strains were either resistant or multiresistant, particularly towards cefuroxime, methicillin, and ceftazidime. Diabetic patients seem to be more prone to B. cereus infections than healthy individuals. It would be greatly beneficial to understand and recognize the prevalence of microorganisms and their resistance patterns for better outcome in ocular surgeries.

Automated ribotyping and antibiotic resistance determining of Bacillus spp from conjunctiva of diabetic patients. We aimed to characterize the phenotype and genotype of Bacillus spp isolated from diabetic patients' eyes, by studying the drug sensitivity patterns with a disc-diffusion method. Fifty eyes of 25 patients with type II diabetes mellitus, with at least 10 years of diabetes history, were included in the study. We analyzed the eyes for the presence of Bacillus spp.; presumptive isolates were identified by morphological, and biochemical tests, and confirmed by the VITEK system. Automated EcoRI ribotyping was performed with a RiboPrinter(®) Microbial Characterization System. We determined the antibiotic resistance of the isolates by the Kirby-Bauer disc diffusion test. Seven out of 25 patients were on insulin treatment; 7 on oral anti-diabetic medication; and 11 on combination therapy of insulin and oral medications. Among the 28 Bacillus spp isolates, 14 were B. cereus, 11 were B. pumilus, 2 were B. mojavensis and 1 was B. subtilis. Almost all the strains were either resistant or multiresistant, particularly towards cefuroxime, methicillin, and ceftazidime. Diabetic patients seem to be more prone to B. cereus infections than healthy individuals. It would be greatly beneficial to understand and recognize the prevalence of microorganisms and their resistance patterns for better outcome in ocular surgeries.

pubmed23n1000_15498

[Standards and quality control points of alginate-based medical devices].

To analyze standards of alginate-based medical devices at home and abroad, and to emphasize key issues of quality control that should be concerned about. Based on investigation of alginate application in medical devices and alginate-related medical standards, alginate-related technical indicators and quality control points were comprehensively analyzed. With the rapid development of alginate-based medical materials and medical devices, the relevant standards at home and abroad have been elaborated on the basic technical indicators and detection methods. In addition to Chinese Pharmacopoeia, China has issued one alginate standard for tissue engineering and three alginate related product standards. Considering the special physical and chemical properties of alginate, researchers also need to focus on the sterilization method, expiry date, molecular weight, and ratio of α-L-guluronic acid to β-D-mannuronic acid of alginate, and impurity content.

[Standards and quality control points of alginate-based medical devices]. To analyze standards of alginate-based medical devices at home and abroad, and to emphasize key issues of quality control that should be concerned about. Based on investigation of alginate application in medical devices and alginate-related medical standards, alginate-related technical indicators and quality control points were comprehensively analyzed. With the rapid development of alginate-based medical materials and medical devices, the relevant standards at home and abroad have been elaborated on the basic technical indicators and detection methods. In addition to Chinese Pharmacopoeia, China has issued one alginate standard for tissue engineering and three alginate related product standards. Considering the special physical and chemical properties of alginate, researchers also need to focus on the sterilization method, expiry date, molecular weight, and ratio of α-L-guluronic acid to β-D-mannuronic acid of alginate, and impurity content.

pubmed23n0279_13871

Abdominal body mass distribution and elevated blood pressure are associated with increased risk of death from cardiovascular diseases and cancer in middle-aged men. The results of a 15- to 20-year follow-up in the Paris prospective study I.

The associations of blood pressure (BP), body mass index (BMI) and iliac-to-thigh circumference index (CI) with total, cardiovascular and cancer mortality were investigated in 7312 middle-aged men, initially free of coronary heart disease, known cancer and not treated for hypertension, who have been followed for 15 to 20 years in the Paris prospective study I. Using Cox survival regression analysis, total mortality (1208 deaths) was found to be highest in relatively lean men (BMI &lt; 24.4 kg/m2) with elevated blood pressure (mean BP &gt; or = 96 mmHg) and central pattern of body mass distribution (CI &gt; 1.82). Cancer causes accounted for a large proportion of the increased mortality risk. In parallel, mortality from cardiovascular diseases increased independently with blood pressure and iliac-to-thigh circumference index, but increased with body mass index only in men with low blood pressure (mean BP &lt; 96 mmHg). Taking into account serum cholesterol and cigarette smoking levels as covariates and excluding deaths occurring 5 to 10 years after the examination only slightly attenuated the intensity of this pattern of association.

Abdominal body mass distribution and elevated blood pressure are associated with increased risk of death from cardiovascular diseases and cancer in middle-aged men. The results of a 15- to 20-year follow-up in the Paris prospective study I. The associations of blood pressure (BP), body mass index (BMI) and iliac-to-thigh circumference index (CI) with total, cardiovascular and cancer mortality were investigated in 7312 middle-aged men, initially free of coronary heart disease, known cancer and not treated for hypertension, who have been followed for 15 to 20 years in the Paris prospective study I. Using Cox survival regression analysis, total mortality (1208 deaths) was found to be highest in relatively lean men (BMI &lt; 24.4 kg/m2) with elevated blood pressure (mean BP &gt; or = 96 mmHg) and central pattern of body mass distribution (CI &gt; 1.82). Cancer causes accounted for a large proportion of the increased mortality risk. In parallel, mortality from cardiovascular diseases increased independently with blood pressure and iliac-to-thigh circumference index, but increased with body mass index only in men with low blood pressure (mean BP &lt; 96 mmHg). Taking into account serum cholesterol and cigarette smoking levels as covariates and excluding deaths occurring 5 to 10 years after the examination only slightly attenuated the intensity of this pattern of association.

pubmed23n0719_11634

Evaluation of exercise capacity with cardiopulmonary exercise testing and BNP levels in adult patients with single or systemic right ventricles.

The aim of the study was to evaluate exercise capacity using cardiopulmonary exercise test (CpET) and serum B-type natriuretic peptide (BNP) levels in patients with single or systemic right ventricles. The study group included 40 patients (16 males) - 17 with transposition of the great arteries after Senning operation, 13 with corrected transposition of the great arteries and 10 with single ventricle after Fontan operation, aged 19-55 years (mean 28.8 ±9.5 years). The control group included 22 healthy individuals (10 males) aged 23-49 years (mean 30.6 ±6.1 years). The majority of patients reported good exercise tolerance - accordingly 27 were classified in NYHA class I (67.5%), 12 (30%) in class II, and only 1 (0.5%) in class III. Cardiopulmonary exercise test revealed significantly lower exercise capacity in study patients than in control subjects. In the study vs. control group VO(2max) was 21.7 ±5.9 vs. 34.2 ±7.4 ml/kg/min (p = 0.00001), maximum heart rate at peak exercise (HRmax) 152.5 ±32.3 vs. 187.2 ±15.6 bpm (p = 0.00001), VE/VCO(2) slope 34.8 ±7.1 vs. 25.7 ±3.2 (p = 0.00001), forced vital capacity (FVC) 3.7 ±0.9l vs. 4.6 ±0.3 (p = 0.03), forced expiratory volume in 1 s (FEV(1)) 3.0 ±0.7 vs. 3.7 ±0.9l (p = 0.0002) respectively. Serum BNP concentrations were higher in study patients than in control subjects; 71.8 ±74.4 vs. 10.7 ±8.1 (pg/ml) respectively (p = 0.00001). No significant correlations between BNP levels and CpET parameters were found. Patients with a morphological right ventricle serving the systemic circulation and those with common ventricle physiology after Fontan operation show markedly reduced exercise capacity. They are also characterized by higher serum BNP concentrations, which do not however correlate with CpET parameters.

Evaluation of exercise capacity with cardiopulmonary exercise testing and BNP levels in adult patients with single or systemic right ventricles. The aim of the study was to evaluate exercise capacity using cardiopulmonary exercise test (CpET) and serum B-type natriuretic peptide (BNP) levels in patients with single or systemic right ventricles. The study group included 40 patients (16 males) - 17 with transposition of the great arteries after Senning operation, 13 with corrected transposition of the great arteries and 10 with single ventricle after Fontan operation, aged 19-55 years (mean 28.8 ±9.5 years). The control group included 22 healthy individuals (10 males) aged 23-49 years (mean 30.6 ±6.1 years). The majority of patients reported good exercise tolerance - accordingly 27 were classified in NYHA class I (67.5%), 12 (30%) in class II, and only 1 (0.5%) in class III. Cardiopulmonary exercise test revealed significantly lower exercise capacity in study patients than in control subjects. In the study vs. control group VO(2max) was 21.7 ±5.9 vs. 34.2 ±7.4 ml/kg/min (p = 0.00001), maximum heart rate at peak exercise (HRmax) 152.5 ±32.3 vs. 187.2 ±15.6 bpm (p = 0.00001), VE/VCO(2) slope 34.8 ±7.1 vs. 25.7 ±3.2 (p = 0.00001), forced vital capacity (FVC) 3.7 ±0.9l vs. 4.6 ±0.3 (p = 0.03), forced expiratory volume in 1 s (FEV(1)) 3.0 ±0.7 vs. 3.7 ±0.9l (p = 0.0002) respectively. Serum BNP concentrations were higher in study patients than in control subjects; 71.8 ±74.4 vs. 10.7 ±8.1 (pg/ml) respectively (p = 0.00001). No significant correlations between BNP levels and CpET parameters were found. Patients with a morphological right ventricle serving the systemic circulation and those with common ventricle physiology after Fontan operation show markedly reduced exercise capacity. They are also characterized by higher serum BNP concentrations, which do not however correlate with CpET parameters.

pubmed23n0011_3766

Prospective analysis of valvular replacement without venting the left ventricle.

The present study is a prospective analysis of a series of consecutive valve replacement and combined (valvular plus coronary artery) operations performed without left ventricular decompression. Results indicate that left ventricular venting (direct or indirect) is unnecessary provided that cardioplegic arrest is used, that the venous line is positioned in the right atrium, and that left-sided pressures are monitored. Left ventricular distention, as measured by left ventricular pressure recording, did not occur; air embolism, as determined clinically and by electroencephalographic (EEG) monitoring, was not encountered. The routine use of a left ventricular vent has been abandoned in most intracardiac operations at our center.

Prospective analysis of valvular replacement without venting the left ventricle. The present study is a prospective analysis of a series of consecutive valve replacement and combined (valvular plus coronary artery) operations performed without left ventricular decompression. Results indicate that left ventricular venting (direct or indirect) is unnecessary provided that cardioplegic arrest is used, that the venous line is positioned in the right atrium, and that left-sided pressures are monitored. Left ventricular distention, as measured by left ventricular pressure recording, did not occur; air embolism, as determined clinically and by electroencephalographic (EEG) monitoring, was not encountered. The routine use of a left ventricular vent has been abandoned in most intracardiac operations at our center.

pubmed23n0876_20473

A mitochondrial DNA hypomorph of cytochrome oxidase specifically impairs male fertility in Drosophila melanogaster.

Due to their strict maternal inheritance in most animals and plants, mitochondrial genomes are predicted to accumulate mutations that are beneficial or neutral in females but harmful in males. Although a few male-harming mtDNA mutations have been identified, consistent with this 'Mother's Curse', their effect on females has been largely unexplored. Here, we identify COII(G177S), a mtDNA hypomorph of cytochrome oxidase II, which specifically impairs male fertility due to defects in sperm development and function without impairing other male or female functions. COII(G177S) represents one of the clearest examples of a 'male-harming' mtDNA mutation in animals and suggest that the hypomorphic mtDNA mutations like COII(G177S) might specifically impair male gametogenesis. Intriguingly, some D. melanogaster nuclear genetic backgrounds can fully rescue COII(G177S) -associated sterility, consistent with previously proposed models that nuclear genomes can regulate the phenotypic manifestation of mtDNA mutations.

A mitochondrial DNA hypomorph of cytochrome oxidase specifically impairs male fertility in Drosophila melanogaster. Due to their strict maternal inheritance in most animals and plants, mitochondrial genomes are predicted to accumulate mutations that are beneficial or neutral in females but harmful in males. Although a few male-harming mtDNA mutations have been identified, consistent with this 'Mother's Curse', their effect on females has been largely unexplored. Here, we identify COII(G177S), a mtDNA hypomorph of cytochrome oxidase II, which specifically impairs male fertility due to defects in sperm development and function without impairing other male or female functions. COII(G177S) represents one of the clearest examples of a 'male-harming' mtDNA mutation in animals and suggest that the hypomorphic mtDNA mutations like COII(G177S) might specifically impair male gametogenesis. Intriguingly, some D. melanogaster nuclear genetic backgrounds can fully rescue COII(G177S) -associated sterility, consistent with previously proposed models that nuclear genomes can regulate the phenotypic manifestation of mtDNA mutations.

pubmed23n1085_22706

Comparison of lag screws and double Y-shaped miniplates in the fixation of anterior mandibular fractures.

Mandibular fractures constitute about 80.79% of maxillofacial injuries in Alexandria University, either as isolated mandibular fractures or as a part of panfacial fractures. The combination of symphyseal and parasymphyseal fractures represent 47.09% of the total mandibular fractures. To compare the effectiveness of lag screws <ivs</i double Y-shaped miniplates in the fixation of anterior mandibular fractures. This study is a prospective randomized controlled clinical trial, performed on sixteen patients with anterior mandibular fractures. Patients were divided equally into two groups, each consisting of eight patients. Group 1: Underwent open reduction and internal fixation using two lag screws. Group 2: Underwent open reduction and internal fixation using double Y-shaped plates. The following parameters were assessed: operating time in minutes, pain using a visual analog scale, edema, surgical wound healing for signs and symptoms of infection, occlusion status and stability, maximal mouth opening, and sensory nerve function. Cone beam computed tomography was performed at 3 and 6 mo to measure bone density and assess the progression of fracture healing. The study included 13 males (81.3%) and 3 females (18.8%) aged 26 to 45 years (mean age was 35.69 ± 6.01 years). The cause of trauma was road traffic accidents in 10 patients (62.5%), interpersonal violence in 3 patients (18.8%) and other causes in 3 patients (18.8%). The fractures comprised 10 parasymphyseal fractures (62.5%) and 6 symphyseal fractures (37.5%). The values of all parameters were comparable in both groups with no statistically significant difference except for the mean bone density at 3 mo postoperatively which was 946.38 ± 66.29 in group 1 and 830.36 ± 95.53 in group 2 (<iP</i = 0.015). Both lag screws and double Y-shaped miniplates provide favorable means of fixation for mandibular fractures in the anterior region. Fractures fixed with lag screws show greater mean bone density at 3 mo post-operation, indicative of higher primary stability and faster early bone healing. Further studies with larger sample sizes are required to verify these conclusions.

Comparison of lag screws and double Y-shaped miniplates in the fixation of anterior mandibular fractures. Mandibular fractures constitute about 80.79% of maxillofacial injuries in Alexandria University, either as isolated mandibular fractures or as a part of panfacial fractures. The combination of symphyseal and parasymphyseal fractures represent 47.09% of the total mandibular fractures. To compare the effectiveness of lag screws <ivs</i double Y-shaped miniplates in the fixation of anterior mandibular fractures. This study is a prospective randomized controlled clinical trial, performed on sixteen patients with anterior mandibular fractures. Patients were divided equally into two groups, each consisting of eight patients. Group 1: Underwent open reduction and internal fixation using two lag screws. Group 2: Underwent open reduction and internal fixation using double Y-shaped plates. The following parameters were assessed: operating time in minutes, pain using a visual analog scale, edema, surgical wound healing for signs and symptoms of infection, occlusion status and stability, maximal mouth opening, and sensory nerve function. Cone beam computed tomography was performed at 3 and 6 mo to measure bone density and assess the progression of fracture healing. The study included 13 males (81.3%) and 3 females (18.8%) aged 26 to 45 years (mean age was 35.69 ± 6.01 years). The cause of trauma was road traffic accidents in 10 patients (62.5%), interpersonal violence in 3 patients (18.8%) and other causes in 3 patients (18.8%). The fractures comprised 10 parasymphyseal fractures (62.5%) and 6 symphyseal fractures (37.5%). The values of all parameters were comparable in both groups with no statistically significant difference except for the mean bone density at 3 mo postoperatively which was 946.38 ± 66.29 in group 1 and 830.36 ± 95.53 in group 2 (<iP</i = 0.015). Both lag screws and double Y-shaped miniplates provide favorable means of fixation for mandibular fractures in the anterior region. Fractures fixed with lag screws show greater mean bone density at 3 mo post-operation, indicative of higher primary stability and faster early bone healing. Further studies with larger sample sizes are required to verify these conclusions.

pubmed23n1057_7199

Epigenetic Regulation of Dental Pulp Stem Cell Fate.

Epigenetic regulation, mainly involving DNA methylation, histone modification, and noncoding RNAs, affects gene expression without modifying the primary DNA sequence and modulates cell fate. Mesenchymal stem cells derived from dental pulp, also called dental pulp stem cells (DPSCs), exhibit multipotent differentiation capacity and can promote various biological processes, including odontogenesis, osteogenesis, angiogenesis, myogenesis, and chondrogenesis. Over the past decades, increased attention has been attracted by the use of DPSCs in the field of regenerative medicine. According to a series of studies, epigenetic regulation is essential for DPSCs to differentiate into specialized cells. In this review, we summarize the mechanisms involved in the epigenetic regulation of the fate of DPSCs.

Epigenetic Regulation of Dental Pulp Stem Cell Fate. Epigenetic regulation, mainly involving DNA methylation, histone modification, and noncoding RNAs, affects gene expression without modifying the primary DNA sequence and modulates cell fate. Mesenchymal stem cells derived from dental pulp, also called dental pulp stem cells (DPSCs), exhibit multipotent differentiation capacity and can promote various biological processes, including odontogenesis, osteogenesis, angiogenesis, myogenesis, and chondrogenesis. Over the past decades, increased attention has been attracted by the use of DPSCs in the field of regenerative medicine. According to a series of studies, epigenetic regulation is essential for DPSCs to differentiate into specialized cells. In this review, we summarize the mechanisms involved in the epigenetic regulation of the fate of DPSCs.

pubmed23n1035_3309

Polarization and function of tumor-associated macrophages mediate graphene oxide-induced photothermal cancer therapy.

Polarization status of tumor-associated macrophages (TAMs) plays an essential role in tumor growth and invasion. However, emerging treatment like photothermal therapy (PTT), photodynamic therapy (PDT) paid little attention on TAMs. In recent years, photothermal therapy (PTT) has gained immense attention in the anti-tumor strategy field while the effect of PTT on macrophage polarization in a tumor microenvironment has rarely been reported. Here, we used graphene oxide (GO) combined with polyethylene glycol (PEG) as the photothermal material to induce heating effect in macrophages to define its anti-tumor effect in vitro and in vivo. Firstly, we treated the macrophage cell line RAW264.7 with near infrared (NIR) light irradiation and detected their polarization status by flow cytometric and mRNA expression analysis. Following this, we analyzed the migration and invasion ability of an osteosarcoma HOS cell line cultured in a conditioned medium (CM) that contains cytokine generated by macrophages with or without NIR treatment. Finally, we investigated the in vivo effects of NIR-induced macrophage polarization on osteosarcoma growth and invasion. GO-PEG (GP) showed great photothermal effect, thermal stability, and biocompatibility in vitro and in vivo. Photothermal materials can alleviate interleukin-4-induced M2 polarization of macrophages and modulate their anti-tumor capability. Thus, the migration and invasion capabilities of HOS cells were weakened, leading to an anti-tumor effect in a mouse subcutaneous tumor model. In conclusion, our study identified PTT treatment as an approach for preventing osteosarcoma invasion by inhibition of M2 polarization.

Polarization and function of tumor-associated macrophages mediate graphene oxide-induced photothermal cancer therapy. Polarization status of tumor-associated macrophages (TAMs) plays an essential role in tumor growth and invasion. However, emerging treatment like photothermal therapy (PTT), photodynamic therapy (PDT) paid little attention on TAMs. In recent years, photothermal therapy (PTT) has gained immense attention in the anti-tumor strategy field while the effect of PTT on macrophage polarization in a tumor microenvironment has rarely been reported. Here, we used graphene oxide (GO) combined with polyethylene glycol (PEG) as the photothermal material to induce heating effect in macrophages to define its anti-tumor effect in vitro and in vivo. Firstly, we treated the macrophage cell line RAW264.7 with near infrared (NIR) light irradiation and detected their polarization status by flow cytometric and mRNA expression analysis. Following this, we analyzed the migration and invasion ability of an osteosarcoma HOS cell line cultured in a conditioned medium (CM) that contains cytokine generated by macrophages with or without NIR treatment. Finally, we investigated the in vivo effects of NIR-induced macrophage polarization on osteosarcoma growth and invasion. GO-PEG (GP) showed great photothermal effect, thermal stability, and biocompatibility in vitro and in vivo. Photothermal materials can alleviate interleukin-4-induced M2 polarization of macrophages and modulate their anti-tumor capability. Thus, the migration and invasion capabilities of HOS cells were weakened, leading to an anti-tumor effect in a mouse subcutaneous tumor model. In conclusion, our study identified PTT treatment as an approach for preventing osteosarcoma invasion by inhibition of M2 polarization.

pubmed23n0647_8113

Electro-acupuncture to prevent prolonged postoperative ileus: a randomized clinical trial.

To examine whether acupuncture can prevent prolonged postoperative ileus (PPOI) after intraperitoneal surgery for colon cancer. Ninety patients were recruited from the Fudan University Cancer Hospital, Shanghai, China. After surgery, patients were randomized to receive acupuncture (once daily, starting on postoperative day 1, for up to six consecutive days) or usual care. PPOI was defined as an inability to pass flatus or have a bowel movement by 96 h after surgery. The main outcomes were time to first flatus, time to first bowel movement, and electrogastroenterography. Secondary outcomes were quality of life (QOL) measures, including pain, nausea, insomnia, abdominal distension/fullness, and sense of well-being. No significant differences in PPOI on day 4 (P = 0.71) or QOL measures were found between the groups. There were also no group differences when the data were analyzed by examining those whose PPOI had resolved by day 5 (P = 0.69) or day 6 (P = 0.88). No adverse events related to acupuncture were reported. Acupuncture did not prevent PPOI and was not useful for treating PPOI once it had developed in this population.

Electro-acupuncture to prevent prolonged postoperative ileus: a randomized clinical trial. To examine whether acupuncture can prevent prolonged postoperative ileus (PPOI) after intraperitoneal surgery for colon cancer. Ninety patients were recruited from the Fudan University Cancer Hospital, Shanghai, China. After surgery, patients were randomized to receive acupuncture (once daily, starting on postoperative day 1, for up to six consecutive days) or usual care. PPOI was defined as an inability to pass flatus or have a bowel movement by 96 h after surgery. The main outcomes were time to first flatus, time to first bowel movement, and electrogastroenterography. Secondary outcomes were quality of life (QOL) measures, including pain, nausea, insomnia, abdominal distension/fullness, and sense of well-being. No significant differences in PPOI on day 4 (P = 0.71) or QOL measures were found between the groups. There were also no group differences when the data were analyzed by examining those whose PPOI had resolved by day 5 (P = 0.69) or day 6 (P = 0.88). No adverse events related to acupuncture were reported. Acupuncture did not prevent PPOI and was not useful for treating PPOI once it had developed in this population.

pubmed23n0879_2341

A stochastic model of the bovine spongiform encephalopathy epidemic in Canada.

Bovine spongiform encephalopathy (BSE) appeared in the United Kingdom in the mid 1980s, and has been attributed to the use of meat and bone meal (MBM) in cattle feed contaminated with a scrapie-like agent. Import of infectious materials from a country where BSE has occurred is believed to be the major factor underlying the spread of the BSE epidemic to other countries. This study presents a new stochastic model developed to estimate risk of BSE from importation of cattle infected with the BSE agent. The model describes the propagation of the BSE agent through the Canadian cattle herd through rendering and feeding processes, following importation of cattle with infectious prions. This model was used estimate the annual number of newly infected animals each year over the period 1980-2019. Model predictions suggested that the number of BSE infections in Canada might have been approximately 40-fold greater than the actual number of clinically diagnosed cases. Under complete compliance with the 2007 ban on feeding MBM, this model further predicts that BSE is disappearing from the Canadian cattle system. A series of sensitivity analyses was also conducted to test the robustness of model predictions to alternative assumptions about factors affecting the evolution of the Canadian BSE epidemic.

A stochastic model of the bovine spongiform encephalopathy epidemic in Canada. Bovine spongiform encephalopathy (BSE) appeared in the United Kingdom in the mid 1980s, and has been attributed to the use of meat and bone meal (MBM) in cattle feed contaminated with a scrapie-like agent. Import of infectious materials from a country where BSE has occurred is believed to be the major factor underlying the spread of the BSE epidemic to other countries. This study presents a new stochastic model developed to estimate risk of BSE from importation of cattle infected with the BSE agent. The model describes the propagation of the BSE agent through the Canadian cattle herd through rendering and feeding processes, following importation of cattle with infectious prions. This model was used estimate the annual number of newly infected animals each year over the period 1980-2019. Model predictions suggested that the number of BSE infections in Canada might have been approximately 40-fold greater than the actual number of clinically diagnosed cases. Under complete compliance with the 2007 ban on feeding MBM, this model further predicts that BSE is disappearing from the Canadian cattle system. A series of sensitivity analyses was also conducted to test the robustness of model predictions to alternative assumptions about factors affecting the evolution of the Canadian BSE epidemic.

pubmed23n0767_9725

MTA resorption and periradicular healing in an open-apex incisor: A case report.

This case report describes the periradicular healing and resorption of an unintentional extrusion of mineral trioxide aggregate (MTA) in an open-apex central incisor. A 22-year old female with a symptomatic open-apex right maxillary central incisor associated with a periradicular lesion was referred for evaluation and treatment. After chemomechanical debridement, the apical third of the root canal was filled with MTA to create an apical plug. Postoperative radiographs showed the extrusion of MTA into the periradicular lesion. The tooth was then restored with a post and crown. At the 2-year follow-up, the tooth was asymptomatic and radiographs revealed complete healing of the periradicular area. At the 7-year follow-up, complete resorption of the extruded MTA was evident. The results of this case study indicate that complete resorption of extruded MTA is possible in the long term; however, the extrusion of MTA in open-apex tooth should still be avoided.

MTA resorption and periradicular healing in an open-apex incisor: A case report. This case report describes the periradicular healing and resorption of an unintentional extrusion of mineral trioxide aggregate (MTA) in an open-apex central incisor. A 22-year old female with a symptomatic open-apex right maxillary central incisor associated with a periradicular lesion was referred for evaluation and treatment. After chemomechanical debridement, the apical third of the root canal was filled with MTA to create an apical plug. Postoperative radiographs showed the extrusion of MTA into the periradicular lesion. The tooth was then restored with a post and crown. At the 2-year follow-up, the tooth was asymptomatic and radiographs revealed complete healing of the periradicular area. At the 7-year follow-up, complete resorption of the extruded MTA was evident. The results of this case study indicate that complete resorption of extruded MTA is possible in the long term; however, the extrusion of MTA in open-apex tooth should still be avoided.

pubmed23n1127_19628

Return to sport post-knee arthroplasty : an umbrella review for consensus guidelines.

Return to sport following undergoing total (TKA) and unicompartmental knee arthroplasty (UKA) has been researched with meta-analyses and systematic reviews of varying quality. The aim of this study is to create an umbrella review to consolidate the data into consensus guidelines for returning to sports following TKA and UKA. Systematic reviews and meta-analyses written between 2010 and 2020 were systematically searched. Studies were independently screened by two reviewers and methodology quality was assessed. Variables for analysis included objective classification of which sports are safe to participate in postoperatively, time to return to sport, prognostic indicators of returning, and reasons patients do not. A total of 410 articles were found, including 58 duplicates. Seven articles meeting inclusion criteria reported that 34% to 100% of patients who underwent TKA or UKA were able to return to sports at 13 weeks and 12 weeks respectively, with UKA patients more likely to do so. Prior experience with the sport was the most significant prognostic indicator for return. These patients were likely to participate in low-impact sports, particularly walking, cycling, golf, and swimming. Moderate-impact sport participation, such as doubles tennis and skiing, may be considered on a case-by-case basis considering the patient's prior experience. There is insufficient long-term data on the risks to return to high-impact sport, such as decreased implant survivorship. There is a consensus that patients can return to low-impact sports following TKA or UKA. Return to moderate-impact sport was dependent on a case-by-case basis, with emphasis on the patient's prior experience in the sport. Return to high-impact sports was not supported. Patients undergoing UKA return to sport one week sooner and with more success than TKA. Future studies are needed to assess long-term outcomes following return to high-impact sports to establish evidence-based recommendations. This review summarizes all available data for the most up-to-date and evidence-based guidelines for returning to sport following TKA and UKA to replace guidelines based on subjective physician survey data. Cite this article: <iBone Jt Open</i 2022;3(3):245-251.

Return to sport post-knee arthroplasty : an umbrella review for consensus guidelines. Return to sport following undergoing total (TKA) and unicompartmental knee arthroplasty (UKA) has been researched with meta-analyses and systematic reviews of varying quality. The aim of this study is to create an umbrella review to consolidate the data into consensus guidelines for returning to sports following TKA and UKA. Systematic reviews and meta-analyses written between 2010 and 2020 were systematically searched. Studies were independently screened by two reviewers and methodology quality was assessed. Variables for analysis included objective classification of which sports are safe to participate in postoperatively, time to return to sport, prognostic indicators of returning, and reasons patients do not. A total of 410 articles were found, including 58 duplicates. Seven articles meeting inclusion criteria reported that 34% to 100% of patients who underwent TKA or UKA were able to return to sports at 13 weeks and 12 weeks respectively, with UKA patients more likely to do so. Prior experience with the sport was the most significant prognostic indicator for return. These patients were likely to participate in low-impact sports, particularly walking, cycling, golf, and swimming. Moderate-impact sport participation, such as doubles tennis and skiing, may be considered on a case-by-case basis considering the patient's prior experience. There is insufficient long-term data on the risks to return to high-impact sport, such as decreased implant survivorship. There is a consensus that patients can return to low-impact sports following TKA or UKA. Return to moderate-impact sport was dependent on a case-by-case basis, with emphasis on the patient's prior experience in the sport. Return to high-impact sports was not supported. Patients undergoing UKA return to sport one week sooner and with more success than TKA. Future studies are needed to assess long-term outcomes following return to high-impact sports to establish evidence-based recommendations. This review summarizes all available data for the most up-to-date and evidence-based guidelines for returning to sport following TKA and UKA to replace guidelines based on subjective physician survey data. Cite this article: <iBone Jt Open</i 2022;3(3):245-251.

pubmed23n0261_14546

The relative contribution of dental services to the changes and geographical variations in caries status of 5- and 12-year-old children in England and Wales in the 1980s.

The aim of this study was to assess the relative contribution of dental services to the changes and geographical variations in caries status of 5- and 12-year-old children in England and Wales in the 1980s. A secondary aim was to assess the association between caries experience and social factors. An ecological study analysing data at a district health authority level in England and Wales was conducted. Twenty-eight, 43 and 34 per cent of the variations in 5-year-old dmft in 1985, 1987 and 1989 respectively were explained by dental service activity indicators and 53, 62 and 57 per cent by social factors. Twenty-nine per cent of the variation in 12-year-old DMFT in 1988 was explained by dental service activity indicators and 46 per cent by social factors.

The relative contribution of dental services to the changes and geographical variations in caries status of 5- and 12-year-old children in England and Wales in the 1980s. The aim of this study was to assess the relative contribution of dental services to the changes and geographical variations in caries status of 5- and 12-year-old children in England and Wales in the 1980s. A secondary aim was to assess the association between caries experience and social factors. An ecological study analysing data at a district health authority level in England and Wales was conducted. Twenty-eight, 43 and 34 per cent of the variations in 5-year-old dmft in 1985, 1987 and 1989 respectively were explained by dental service activity indicators and 53, 62 and 57 per cent by social factors. Twenty-nine per cent of the variation in 12-year-old DMFT in 1988 was explained by dental service activity indicators and 46 per cent by social factors.

pubmed23n0074_12563

Coronal biparietal diameter. A reliable alternative to the traditional transverse biparietal diameter.

The traditional ultrasonically derived fetal biparietal diameter (BPD) is important in the determination of fetal age, weight, and growth. This measure cannot be obtained, however, with direct occiput anterior or posterior or with a head in the pelvis. This study looked at the BPD utilizing coronal (anatomically at right angles to the traditional BPD through the thalami, third ventricle, and cavum septum pellucidum) rather than transverse section through the fetal head. In 265 patients at 14 to 41 weeks' gestation, simultaneous measurement of the coronal and transverse BPDs revealed that they were interchangeable (r = 0.99). The major drawback of the coronal BPD is the inability to obtain a head circumference because of the absence of the occipitofrontal diameter.

Coronal biparietal diameter. A reliable alternative to the traditional transverse biparietal diameter. The traditional ultrasonically derived fetal biparietal diameter (BPD) is important in the determination of fetal age, weight, and growth. This measure cannot be obtained, however, with direct occiput anterior or posterior or with a head in the pelvis. This study looked at the BPD utilizing coronal (anatomically at right angles to the traditional BPD through the thalami, third ventricle, and cavum septum pellucidum) rather than transverse section through the fetal head. In 265 patients at 14 to 41 weeks' gestation, simultaneous measurement of the coronal and transverse BPDs revealed that they were interchangeable (r = 0.99). The major drawback of the coronal BPD is the inability to obtain a head circumference because of the absence of the occipitofrontal diameter.

pubmed23n1105_13469

Context Modulates Attention to Faces in Dynamic Social Scenes in Children and Adults with Autism Spectrum Disorder.

Individuals with autism spectrum disorder (ASD) have been found to view social scenes differently compared to typically developing (TD) peers, but results can vary depending on context and age. We used eye-tracking in children and adults (age 6-63) to assess allocation of visual attention in a dynamic social orientation paradigm previously used only in younger children. The ASD group (n = 94) looked less at the actor's face compared to TD (n = 38) when they were engaged in activity (mean percentage of looking time, ASD = 30.7% vs TD = 34.9%; Cohen's d = 0.56; p value &lt; 0.03) or looking at a moving toy (24.5% vs 33.2%; d = 0.65; p value &lt; 0.001). Findings indicate that there are qualitative differences in allocation of visual attention to social stimuli across ages in ASD.ClinicalTrials.gov identifier: NCT02668991.

Context Modulates Attention to Faces in Dynamic Social Scenes in Children and Adults with Autism Spectrum Disorder. Individuals with autism spectrum disorder (ASD) have been found to view social scenes differently compared to typically developing (TD) peers, but results can vary depending on context and age. We used eye-tracking in children and adults (age 6-63) to assess allocation of visual attention in a dynamic social orientation paradigm previously used only in younger children. The ASD group (n = 94) looked less at the actor's face compared to TD (n = 38) when they were engaged in activity (mean percentage of looking time, ASD = 30.7% vs TD = 34.9%; Cohen's d = 0.56; p value &lt; 0.03) or looking at a moving toy (24.5% vs 33.2%; d = 0.65; p value &lt; 0.001). Findings indicate that there are qualitative differences in allocation of visual attention to social stimuli across ages in ASD.ClinicalTrials.gov identifier: NCT02668991.

pubmed23n0969_6107

Detection of patients at high risk for nonocclusive mesenteric ischemia after cardiovascular surgery.

Nonocclusive mesenteric ischemia (NOMI) is a rare but life-threatening complication after cardiovascular surgery. Early diagnosis and treatment is essential for a chance to cure. The aim of this study is to identify the independent risk factors for NOMI based on the evaluation of 12 cases of NOMI after cardiovascular surgery. We retrospectively analyzed 12 patients with NOMI and 674 other patients without NOMI who underwent cardiovascular surgery in our hospital. We reviewed the clinical data on NOMI patients, including their characteristics and the clinical course. In addition, we performed a statistical comparison of each factor from both NOMI and non-NOMI groups to identify the independent risk factors for NOMI. The median duration between the cardiac surgery and the diagnosis of NOMI was 14.0 (10.3-20.3) days. The in-hospital mortality of NOMI patients was 75.0%. Age (p &lt; 0.05), peripheral arterial disease (p &lt; 0.001), postoperative hemodialysis (p &lt; 0.001), intraaortic balloon pump (p &lt; 0.05), norepinephrine (NOE) &gt; 0.10γ (p &lt; 0.0001), percutaneous cardiopulmonary support (p &lt; 0.001), sepsis (p &lt; 0.05), loss of sinus rhythm (p &lt; 0.05), prolonged ventilation (p &lt; 0.0001), and resternotomy for bleeding (p &lt; 0.05) showed significant differences between NOMI and non-NOMI groups. In the multivariate logistic regression model, prolonged ventilation [odds ratio (OR) = 18.1, p &lt; 0.001] and NOE &gt; 0.10 μg/kg/min (OR = 130.0, p &lt; 0.0001) were detected as independent risk factors for NOMI. We have identified the risk factors for NOMI based on the evaluation of the 12 cases of NOMI after cardiovascular surgery. This result may be useful in predicting NOMI, which is considered difficult in clinical practice. For the patient with suspected of NOMI who has these risk factors, early CT scan and surgical exploration should be performed without delay.

Detection of patients at high risk for nonocclusive mesenteric ischemia after cardiovascular surgery. Nonocclusive mesenteric ischemia (NOMI) is a rare but life-threatening complication after cardiovascular surgery. Early diagnosis and treatment is essential for a chance to cure. The aim of this study is to identify the independent risk factors for NOMI based on the evaluation of 12 cases of NOMI after cardiovascular surgery. We retrospectively analyzed 12 patients with NOMI and 674 other patients without NOMI who underwent cardiovascular surgery in our hospital. We reviewed the clinical data on NOMI patients, including their characteristics and the clinical course. In addition, we performed a statistical comparison of each factor from both NOMI and non-NOMI groups to identify the independent risk factors for NOMI. The median duration between the cardiac surgery and the diagnosis of NOMI was 14.0 (10.3-20.3) days. The in-hospital mortality of NOMI patients was 75.0%. Age (p &lt; 0.05), peripheral arterial disease (p &lt; 0.001), postoperative hemodialysis (p &lt; 0.001), intraaortic balloon pump (p &lt; 0.05), norepinephrine (NOE) &gt; 0.10γ (p &lt; 0.0001), percutaneous cardiopulmonary support (p &lt; 0.001), sepsis (p &lt; 0.05), loss of sinus rhythm (p &lt; 0.05), prolonged ventilation (p &lt; 0.0001), and resternotomy for bleeding (p &lt; 0.05) showed significant differences between NOMI and non-NOMI groups. In the multivariate logistic regression model, prolonged ventilation [odds ratio (OR) = 18.1, p &lt; 0.001] and NOE &gt; 0.10 μg/kg/min (OR = 130.0, p &lt; 0.0001) were detected as independent risk factors for NOMI. We have identified the risk factors for NOMI based on the evaluation of the 12 cases of NOMI after cardiovascular surgery. This result may be useful in predicting NOMI, which is considered difficult in clinical practice. For the patient with suspected of NOMI who has these risk factors, early CT scan and surgical exploration should be performed without delay.

pubmed23n0544_14654

Iron fortification reduces blood lead levels in children in Bangalore, India.

Chronic lead poisoning and iron deficiency are concentrated in urban children from lower socioeconomic strata, and both impair neurocognitive development. Our study objective was to determine if iron fortification reduces blood lead levels in urban, lead-exposed, iron-deficient children in Bangalore, India. A randomized, double-blind, controlled school-based feeding trial was done in 5- to 13-year-old iron-deficient children (n = 186). At baseline, a high prevalence of lead poisoning was found in the younger children. Subsequently, all 5- to 9-year-old children participating in the trial (n = 134) were followed to determine if iron fortification would affect their blood lead levels. Children were dewormed and fed 6 days/week for 16 weeks either an iron-fortified rice meal (approximately 15 mg of iron per day as ferric pyrophosphate) or an identical control meal without added iron. Feeding was directly supervised and compliance monitored. Hemoglobin, serum ferritin, C-reactive protein, transferrin receptor, zinc protoporphyrin, and blood lead concentrations were measured. The prevalence of iron deficiency was significantly reduced in the iron group (from 70% to 28%) compared with the control group (76% to 55%). There was a significant decrease in median blood lead concentration in the iron group compared with the control group. The prevalence of blood lead levels &gt; or =10 microg/dL was significantly reduced in the iron group (from 65% to 29%) compared with the control group (68% to 55%). Our findings suggest providing iron in a fortified food to lead-exposed children may reduce chronic lead intoxication. Iron fortification may be an effective and sustainable strategy to accompany environmental lead abatement.

Iron fortification reduces blood lead levels in children in Bangalore, India. Chronic lead poisoning and iron deficiency are concentrated in urban children from lower socioeconomic strata, and both impair neurocognitive development. Our study objective was to determine if iron fortification reduces blood lead levels in urban, lead-exposed, iron-deficient children in Bangalore, India. A randomized, double-blind, controlled school-based feeding trial was done in 5- to 13-year-old iron-deficient children (n = 186). At baseline, a high prevalence of lead poisoning was found in the younger children. Subsequently, all 5- to 9-year-old children participating in the trial (n = 134) were followed to determine if iron fortification would affect their blood lead levels. Children were dewormed and fed 6 days/week for 16 weeks either an iron-fortified rice meal (approximately 15 mg of iron per day as ferric pyrophosphate) or an identical control meal without added iron. Feeding was directly supervised and compliance monitored. Hemoglobin, serum ferritin, C-reactive protein, transferrin receptor, zinc protoporphyrin, and blood lead concentrations were measured. The prevalence of iron deficiency was significantly reduced in the iron group (from 70% to 28%) compared with the control group (76% to 55%). There was a significant decrease in median blood lead concentration in the iron group compared with the control group. The prevalence of blood lead levels &gt; or =10 microg/dL was significantly reduced in the iron group (from 65% to 29%) compared with the control group (68% to 55%). Our findings suggest providing iron in a fortified food to lead-exposed children may reduce chronic lead intoxication. Iron fortification may be an effective and sustainable strategy to accompany environmental lead abatement.

pubmed23n0348_6503

Effects of dopamine D(1-like) and D(2-like) agonists in rats that self-administer cocaine.

The reinforcing effects of D(1-like) and D(2-like) agonists, and their capacity to modify cocaine self-administration, were compared in rats with extensive cocaine self-administration experience. Cocaine (0.01-1.0 mg i.v.) dose-dependently maintained responding under a fixed ratio (FR) 5 schedule of reinforcement, and an inverted U-shaped function characterized the relationship between unit dose and self-administration behavior. When substituted for cocaine, the D(1-like) agonists SKF 82958 (0.001-0.032 mg i.v.) and SKF 77434 (0.001-0.1 mg i.v.) did not maintain responding above levels observed during saline substitution. In contrast, the D(2-like) agonists quinelorane (0.001-0.1 mg i.v.) and 7-hydroxy-dipropylaminotetralin (7-OH-DPAT; 0.01-0.32 mg i.v.) reliably maintained i.v. self-administration behavior that was characterized by inverted U-shaped dose-effect functions. Pretreatment with the D(1-like) agonists SKF 82958 and SKF 77434 (0.1-1.0 mg/kg i.p.) shifted the dose-effect function for cocaine self-administration downward, whereas pretreatment with the D(2-like) agonists quinelorane (0.01 mg/kg i.p.) and 7-OH-DPAT (0.32-1.0 mg/kg i.p.) shifted the cocaine dose-effect function to the left. Effects of D(1-like) and D(2-like) agonists on patterns of responding maintained by cocaine (0.32 mg i.v.) also differed: D(1-like) agonists increased the latency to the first response but did not otherwise alter patterns of cocaine self-administration, whereas D(2-like) agonists increased the intervals between self-administered cocaine injections. The results suggest that D(2-like) agonists, but not D(1-like) agonists, have prominent reinforcing effects and enhance the effects of self-administered cocaine in rats with extensive cocaine self-administration experience. Consequently, D(2) receptor-related neuronal mechanisms may be especially important in mediating the abuse-related effects of cocaine.

Effects of dopamine D(1-like) and D(2-like) agonists in rats that self-administer cocaine. The reinforcing effects of D(1-like) and D(2-like) agonists, and their capacity to modify cocaine self-administration, were compared in rats with extensive cocaine self-administration experience. Cocaine (0.01-1.0 mg i.v.) dose-dependently maintained responding under a fixed ratio (FR) 5 schedule of reinforcement, and an inverted U-shaped function characterized the relationship between unit dose and self-administration behavior. When substituted for cocaine, the D(1-like) agonists SKF 82958 (0.001-0.032 mg i.v.) and SKF 77434 (0.001-0.1 mg i.v.) did not maintain responding above levels observed during saline substitution. In contrast, the D(2-like) agonists quinelorane (0.001-0.1 mg i.v.) and 7-hydroxy-dipropylaminotetralin (7-OH-DPAT; 0.01-0.32 mg i.v.) reliably maintained i.v. self-administration behavior that was characterized by inverted U-shaped dose-effect functions. Pretreatment with the D(1-like) agonists SKF 82958 and SKF 77434 (0.1-1.0 mg/kg i.p.) shifted the dose-effect function for cocaine self-administration downward, whereas pretreatment with the D(2-like) agonists quinelorane (0.01 mg/kg i.p.) and 7-OH-DPAT (0.32-1.0 mg/kg i.p.) shifted the cocaine dose-effect function to the left. Effects of D(1-like) and D(2-like) agonists on patterns of responding maintained by cocaine (0.32 mg i.v.) also differed: D(1-like) agonists increased the latency to the first response but did not otherwise alter patterns of cocaine self-administration, whereas D(2-like) agonists increased the intervals between self-administered cocaine injections. The results suggest that D(2-like) agonists, but not D(1-like) agonists, have prominent reinforcing effects and enhance the effects of self-administered cocaine in rats with extensive cocaine self-administration experience. Consequently, D(2) receptor-related neuronal mechanisms may be especially important in mediating the abuse-related effects of cocaine.

pubmed23n0821_15094

Effects of Electronic Nicotine Delivery System on Larynx: Experimental Study.

We aimed to assess the effects of electronic nicotine delivery system (ENDS) or also termed electronic cigarette vapor on the laryngeal mucosa of rats. Sixteen female Wistar albino rats were divided into two groups. The study group was exposed to ENDS vapor for 1 hour/day for 4 weeks. The control group was not subjected to any chemical or physical stimulus. The vocal folds of the study and control group rats were evaluated histopathologically by hematoxylin and eosin staining and immunohistochemically by Ki67 staining. Epithelial distribution, inflammation, hyperplasia, and metaplasia were evaluated. Epithelial distribution and inflammation did not differ between the two groups. Two cases of hyperplasia were detected in the study group but there was no hyperplasia in the control group. Four cases of metaplasia were detected in the study group and one case in the control group. Statistical analysis revealed no significant difference between the study and control groups (P = 0.131 and 0.106, respectively). Exposure to ENDS for 4 weeks caused hyperplasia and metaplasia of the laryngeal mucosa of rats but this was not significant statistically. These results implemented that further studies with larger cohort and longer duration are required to evaluate long-term effects.

Effects of Electronic Nicotine Delivery System on Larynx: Experimental Study. We aimed to assess the effects of electronic nicotine delivery system (ENDS) or also termed electronic cigarette vapor on the laryngeal mucosa of rats. Sixteen female Wistar albino rats were divided into two groups. The study group was exposed to ENDS vapor for 1 hour/day for 4 weeks. The control group was not subjected to any chemical or physical stimulus. The vocal folds of the study and control group rats were evaluated histopathologically by hematoxylin and eosin staining and immunohistochemically by Ki67 staining. Epithelial distribution, inflammation, hyperplasia, and metaplasia were evaluated. Epithelial distribution and inflammation did not differ between the two groups. Two cases of hyperplasia were detected in the study group but there was no hyperplasia in the control group. Four cases of metaplasia were detected in the study group and one case in the control group. Statistical analysis revealed no significant difference between the study and control groups (P = 0.131 and 0.106, respectively). Exposure to ENDS for 4 weeks caused hyperplasia and metaplasia of the laryngeal mucosa of rats but this was not significant statistically. These results implemented that further studies with larger cohort and longer duration are required to evaluate long-term effects.

pubmed23n0778_6625

Online exercise for the design and simulation of PCR and PCR-RFLP experiments.

Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism of PCR products (PCR-RFLP) are extensively used molecular biology techniques. An exercise for the design and simulation of PCR and PCR-RFLP experiments will be a useful educational tool. An online PCR and PCR-RFLP exercise has been create that requires users to find the target genes, compare them, design primers, search for restriction endonucleases, and finally to simulate the experiment. Each user of the service is randomly assigned a gene from Escherichia coli; to complete the exercise, users must design an experiment capable of distinguishing among E. coli strains. By applying the experimental procedure to all completely sequenced E. coli, a basic understanding of strain comparison and clustering can also be acquired. Comparison of results obtained in different experiments is also very instructive. The exercise is freely available at http://insilico.ehu.es/edu.

Online exercise for the design and simulation of PCR and PCR-RFLP experiments. Polymerase Chain Reaction (PCR) and Restriction Fragment Length Polymorphism of PCR products (PCR-RFLP) are extensively used molecular biology techniques. An exercise for the design and simulation of PCR and PCR-RFLP experiments will be a useful educational tool. An online PCR and PCR-RFLP exercise has been create that requires users to find the target genes, compare them, design primers, search for restriction endonucleases, and finally to simulate the experiment. Each user of the service is randomly assigned a gene from Escherichia coli; to complete the exercise, users must design an experiment capable of distinguishing among E. coli strains. By applying the experimental procedure to all completely sequenced E. coli, a basic understanding of strain comparison and clustering can also be acquired. Comparison of results obtained in different experiments is also very instructive. The exercise is freely available at http://insilico.ehu.es/edu.

pubmed23n0537_5113

The lipoprotein lipase inhibitor ANGPTL3 is negatively regulated by thyroid hormone.

Whereas the role of thyroid hormone is clearly established in the regulation of cholesterol homeostasis, its involvement in the control of serum triglyceride (TG) levels remains largely debated. Angiopoietin-like proteins 3 and 4 have recently been characterized as potent lipoprotein lipase inhibitors and therefore as important components of plasma triglyceride homeostasis. In the present study, the role of thyroid hormone in the regulation of both ANGPTL4 and ANGPTL3 gene expression was investigated. In vivo studies revealed that thyroid hormone down-regulates ANGPTL3 but not ANGPTL4 gene expression in hypothyroid rats. Using thyroid hormone receptor (TR)-deficient mice, we show that thyroid hormone regulates ANGPTL3 gene expression in a TRbeta-dependent manner. Transfection studies revealed that this inhibition occurs at the transcriptional level in a DNA binding-independent fashion and requires the proximal (-171 to +66) region of the ANGPTL3 gene promoter. Moreover, site-directed mutagenesis experiments indicate that the HNF1 site within this proximal region mediates this TRbeta-dependent repression. Finally, co-transfection studies and electrophoretic mobility shift assays suggest that TRbeta antagonizes the HNF1alpha signaling pathway by inhibiting its transcriptional activity without interfering with its DNA-binding capacity. Taken together, our results lead to the identification of ANGPTL3 as a novel TRbeta target gene and provide a new potential mechanism to explain the hypotriglyceridemic properties of TRbeta agonists in vivo.

The lipoprotein lipase inhibitor ANGPTL3 is negatively regulated by thyroid hormone. Whereas the role of thyroid hormone is clearly established in the regulation of cholesterol homeostasis, its involvement in the control of serum triglyceride (TG) levels remains largely debated. Angiopoietin-like proteins 3 and 4 have recently been characterized as potent lipoprotein lipase inhibitors and therefore as important components of plasma triglyceride homeostasis. In the present study, the role of thyroid hormone in the regulation of both ANGPTL4 and ANGPTL3 gene expression was investigated. In vivo studies revealed that thyroid hormone down-regulates ANGPTL3 but not ANGPTL4 gene expression in hypothyroid rats. Using thyroid hormone receptor (TR)-deficient mice, we show that thyroid hormone regulates ANGPTL3 gene expression in a TRbeta-dependent manner. Transfection studies revealed that this inhibition occurs at the transcriptional level in a DNA binding-independent fashion and requires the proximal (-171 to +66) region of the ANGPTL3 gene promoter. Moreover, site-directed mutagenesis experiments indicate that the HNF1 site within this proximal region mediates this TRbeta-dependent repression. Finally, co-transfection studies and electrophoretic mobility shift assays suggest that TRbeta antagonizes the HNF1alpha signaling pathway by inhibiting its transcriptional activity without interfering with its DNA-binding capacity. Taken together, our results lead to the identification of ANGPTL3 as a novel TRbeta target gene and provide a new potential mechanism to explain the hypotriglyceridemic properties of TRbeta agonists in vivo.

pubmed23n0836_23232

Enhanced x-ray emission from nano-particle doped bacteria.

Recently, it has been greatly appreciated that intense light matter interaction is modified due to the nano- and microstructures in the target by--surface plasmons, laser energy localization scattering etc. Extreme laser intensities produce dense plasmas and collective mechanisms generate energetic electrons, ions and hard x-rays. Recently, it is postulated that the anharmonic electron motion, driven by ultrashort, high-intensity laser pulses, provides a universal mechanism for the laser absorption. Here, we provide the first demonstration of anharmonic-resonance-aided high laser-absorption in a biological system. At intensities of ∼ 10¹⁶⁻¹⁸ W/cm², 40 fs pulses excite a plasma formed with E. coli bacteria. The density-inhomogeneities due to the micro- and nanostructures in the bacterial target increase anharmonic resonance (AHR) heating and result in a 10⁴-fold enhancement in the hard x-ray yield compared to plain solid targets. These observations lead to novel high-energy x-ray sources that have implications to lithography, imaging and medical applications.

Enhanced x-ray emission from nano-particle doped bacteria. Recently, it has been greatly appreciated that intense light matter interaction is modified due to the nano- and microstructures in the target by--surface plasmons, laser energy localization scattering etc. Extreme laser intensities produce dense plasmas and collective mechanisms generate energetic electrons, ions and hard x-rays. Recently, it is postulated that the anharmonic electron motion, driven by ultrashort, high-intensity laser pulses, provides a universal mechanism for the laser absorption. Here, we provide the first demonstration of anharmonic-resonance-aided high laser-absorption in a biological system. At intensities of ∼ 10¹⁶⁻¹⁸ W/cm², 40 fs pulses excite a plasma formed with E. coli bacteria. The density-inhomogeneities due to the micro- and nanostructures in the bacterial target increase anharmonic resonance (AHR) heating and result in a 10⁴-fold enhancement in the hard x-ray yield compared to plain solid targets. These observations lead to novel high-energy x-ray sources that have implications to lithography, imaging and medical applications.

pubmed23n0843_13208

Fatal Insulin Overdoses: Case Report and Update on Testing Methodology.

Suicidal insulin overdoses are an under-recognized and uncommon cause of death, often relying on scene and nonspecific autopsy findings. Here, we present a case report of a fatal exogenous insulin overdose in a patient with type 1 diabetes. In our case, there were no contributory autopsy findings; however, serum analog aspart insulin levels were c. 10× the predicted therapeutic upper limit (4000, reference 6.6-55 uU/mL), which correlated with scene findings. This was specifically determined by a newly developed immunocapture liquid chromatography-tandem mass spectrometry assay, able to discriminate between various synthetic insulin analogs. Total insulin levels by immunoassay were highly elevated on the Siemens Advia Centaur, but not the Roche platforms (4741 vs. 5.2 uU/mL, respectively), showing variable sensitivity of detection within the same analog depending on assay. We discuss the prevalence and features to look for at autopsy in these types of cases. Additionally, analytical options for testing insulin levels, including new methodologies, guidance on collection of samples, as well as an outline of available historical reference range data are discussed.

Fatal Insulin Overdoses: Case Report and Update on Testing Methodology. Suicidal insulin overdoses are an under-recognized and uncommon cause of death, often relying on scene and nonspecific autopsy findings. Here, we present a case report of a fatal exogenous insulin overdose in a patient with type 1 diabetes. In our case, there were no contributory autopsy findings; however, serum analog aspart insulin levels were c. 10× the predicted therapeutic upper limit (4000, reference 6.6-55 uU/mL), which correlated with scene findings. This was specifically determined by a newly developed immunocapture liquid chromatography-tandem mass spectrometry assay, able to discriminate between various synthetic insulin analogs. Total insulin levels by immunoassay were highly elevated on the Siemens Advia Centaur, but not the Roche platforms (4741 vs. 5.2 uU/mL, respectively), showing variable sensitivity of detection within the same analog depending on assay. We discuss the prevalence and features to look for at autopsy in these types of cases. Additionally, analytical options for testing insulin levels, including new methodologies, guidance on collection of samples, as well as an outline of available historical reference range data are discussed.

pubmed23n0394_18490

Counterregulation of chromatin deacetylation and histone deacetylase occupancy at the integrated promoter of human immunodeficiency virus type 1 (HIV-1) by the HIV-1 repressor YY1 and HIV-1 activator Tat.

Repression of human immunodeficiency virus type 1 (HIV-1) transcription may contribute to the establishment or maintenance of proviral quiescence in infected CD4(+) cells. The host factors YY1 and LSF cooperatively recruit histone deacetylase 1 (HDAC1) to the HIV-1 long terminal repeat (LTR) and inhibit transcription. We demonstrate here regulation of occupancy of HDAC1 at a positioned nucleosome (nuc 1) near the transcription start site of integrated LTR. We find that expression of YY1 increases occupancy by HDAC1, decreases acetylation at nuc 1, and downregulates LTR expression. HDAC1 recruitment and histone hypoacetylation were also seen when Tat activation was inhibited by the overexpression of YY1. A YY1 mutant without an HDAC1 interaction domain and incompetent to inhibit LTR activation fails to recruit HDAC1 to LTR or decrease nuc 1 acetylation. Further, expression of a dominant-negative mutant of LSF (dnLSF), which inhibits LSF occupancy and LTR repression, results in acetylation and decreased HDAC1 occupancy at nuc 1. Conversely, exposure of cells to the histone deacetylase inhibitor trichostatin A or activation of LTR expression by HIV-1 Tat results in the displacement of HDAC1 from nuc 1, in association with increased acetylation of histone H4. Recruitment of HDAC1 to the LTR nuc 1 can counteract Tat activation and repress LTR expression. Significantly, when repression is overcome, LTR activation is associated with decreased HDAC1 occupancy. Since the persistence of integrated HIV-1 genomes despite potent suppression of viral replication is a major obstacle for current antiretroviral therapy, strategies to selectively disrupt the quiescence of chromosomal provirus may play a role in the future treatment of AIDS.

Counterregulation of chromatin deacetylation and histone deacetylase occupancy at the integrated promoter of human immunodeficiency virus type 1 (HIV-1) by the HIV-1 repressor YY1 and HIV-1 activator Tat. Repression of human immunodeficiency virus type 1 (HIV-1) transcription may contribute to the establishment or maintenance of proviral quiescence in infected CD4(+) cells. The host factors YY1 and LSF cooperatively recruit histone deacetylase 1 (HDAC1) to the HIV-1 long terminal repeat (LTR) and inhibit transcription. We demonstrate here regulation of occupancy of HDAC1 at a positioned nucleosome (nuc 1) near the transcription start site of integrated LTR. We find that expression of YY1 increases occupancy by HDAC1, decreases acetylation at nuc 1, and downregulates LTR expression. HDAC1 recruitment and histone hypoacetylation were also seen when Tat activation was inhibited by the overexpression of YY1. A YY1 mutant without an HDAC1 interaction domain and incompetent to inhibit LTR activation fails to recruit HDAC1 to LTR or decrease nuc 1 acetylation. Further, expression of a dominant-negative mutant of LSF (dnLSF), which inhibits LSF occupancy and LTR repression, results in acetylation and decreased HDAC1 occupancy at nuc 1. Conversely, exposure of cells to the histone deacetylase inhibitor trichostatin A or activation of LTR expression by HIV-1 Tat results in the displacement of HDAC1 from nuc 1, in association with increased acetylation of histone H4. Recruitment of HDAC1 to the LTR nuc 1 can counteract Tat activation and repress LTR expression. Significantly, when repression is overcome, LTR activation is associated with decreased HDAC1 occupancy. Since the persistence of integrated HIV-1 genomes despite potent suppression of viral replication is a major obstacle for current antiretroviral therapy, strategies to selectively disrupt the quiescence of chromosomal provirus may play a role in the future treatment of AIDS.

pubmed23n0110_8013

Inactive renin and aldosterone in Bartter's syndrome.

Studies of plasma samples of 3 subjects with Bartter's syndrome were compared to 8 subjects with other conditions. Despite high levels of active renin initially, with low levels of inactive renin, addition of either human nephrectomized plasma or sheep substrate not only increased active renin (by at least 3-fold) but also led to the appearance of large quantities of inactive renin (10-20 times the concentration originally present, much greater than the small increase seen with other plasmas). The activated inactive renin after substrate addition possibly had a larger and more variable molecular size (42,000-48,000) than normal inactive renin (42,500-44,500). Renin substrate in Bartter's plasma was present in similar amounts and had a normal or supranormal angiotensin generation rate with exogenous human renin. Bartter's substrate had a similar molecular weight (55,000) to that found in normal human plasma. The agent in the exogenous substrate preparations causing the increase in apparent active and inactive renin was not ultrafiltrable. However, an acidification procedure that destroyed exogenous substrate also removed the renin-increasing effect. Captopril increased renin but not aldosterone, while amiloride increased aldosterone but not renin. Neither agent improved serum potassium significantly in these patients on indomethacin.

Inactive renin and aldosterone in Bartter's syndrome. Studies of plasma samples of 3 subjects with Bartter's syndrome were compared to 8 subjects with other conditions. Despite high levels of active renin initially, with low levels of inactive renin, addition of either human nephrectomized plasma or sheep substrate not only increased active renin (by at least 3-fold) but also led to the appearance of large quantities of inactive renin (10-20 times the concentration originally present, much greater than the small increase seen with other plasmas). The activated inactive renin after substrate addition possibly had a larger and more variable molecular size (42,000-48,000) than normal inactive renin (42,500-44,500). Renin substrate in Bartter's plasma was present in similar amounts and had a normal or supranormal angiotensin generation rate with exogenous human renin. Bartter's substrate had a similar molecular weight (55,000) to that found in normal human plasma. The agent in the exogenous substrate preparations causing the increase in apparent active and inactive renin was not ultrafiltrable. However, an acidification procedure that destroyed exogenous substrate also removed the renin-increasing effect. Captopril increased renin but not aldosterone, while amiloride increased aldosterone but not renin. Neither agent improved serum potassium significantly in these patients on indomethacin.

pubmed23n1157_12426

Eicosapentaenoic Acid Ameliorates Cardiac Fibrosis and Tissue Inflammation in Spontaneously Hypertensive Rats.

Hypertension affects 1 in 3 adults in the United States and leads to left ventricular (LV) concentric hypertrophy, interstitial fibrosis, and increased stiffness. The treatment of cardiac fibrosis remains challenging and empiric. Eicosapentaenoic acid (EPA) is an omega-3 polyunsaturated fatty acid that is highly effective in reducing cardiovascular events in patients and cardiac fibrosis and hypertrophy in animals when administered before pressure overload by promoting the increase of anti-inflammatory M1 macrophages. In this study, we investigated whether EPA mitigates the exacerbation of cardiac remodeling and fibrosis induced by established hypertension, a situation that closely recapitulates a clinical scenario. Twelve-week-old spontaneously hypertensive rats were randomized to eat an EPA-enriched or control diet for 20 weeks. We report that rats eating the EPA-enriched diet exhibited a reduction of interstitial cardiac fibrosis and ameliorated LV diastolic dysfunction despite the continuous increase in blood pressure. However, we found that EPA did not have an impact on cardiac hypertrophy. Interestingly, the EPA diet increased mRNA expression of M2 macrophage marker Mrc1 and interleukin-10 in cardiac tissue. These findings indicated that the antifibrotic effects of EPA are mediated in part by phenotypic polarization of macrophages toward anti-inflammatory M2 macrophages and increases of the anti-inflammatory cytokine, interleukin-10. In summary, EPA prevents the exacerbation of cardiac fibrosis and LV diastolic dysfunction during sustained pressure overload. EPA could represent a novel treatment strategy for hypertensive cardiomyopathy.

Eicosapentaenoic Acid Ameliorates Cardiac Fibrosis and Tissue Inflammation in Spontaneously Hypertensive Rats. Hypertension affects 1 in 3 adults in the United States and leads to left ventricular (LV) concentric hypertrophy, interstitial fibrosis, and increased stiffness. The treatment of cardiac fibrosis remains challenging and empiric. Eicosapentaenoic acid (EPA) is an omega-3 polyunsaturated fatty acid that is highly effective in reducing cardiovascular events in patients and cardiac fibrosis and hypertrophy in animals when administered before pressure overload by promoting the increase of anti-inflammatory M1 macrophages. In this study, we investigated whether EPA mitigates the exacerbation of cardiac remodeling and fibrosis induced by established hypertension, a situation that closely recapitulates a clinical scenario. Twelve-week-old spontaneously hypertensive rats were randomized to eat an EPA-enriched or control diet for 20 weeks. We report that rats eating the EPA-enriched diet exhibited a reduction of interstitial cardiac fibrosis and ameliorated LV diastolic dysfunction despite the continuous increase in blood pressure. However, we found that EPA did not have an impact on cardiac hypertrophy. Interestingly, the EPA diet increased mRNA expression of M2 macrophage marker Mrc1 and interleukin-10 in cardiac tissue. These findings indicated that the antifibrotic effects of EPA are mediated in part by phenotypic polarization of macrophages toward anti-inflammatory M2 macrophages and increases of the anti-inflammatory cytokine, interleukin-10. In summary, EPA prevents the exacerbation of cardiac fibrosis and LV diastolic dysfunction during sustained pressure overload. EPA could represent a novel treatment strategy for hypertensive cardiomyopathy.

pubmed23n1010_21904

Work-related stress and cognitive performance among middle-aged adults: The Brazilian Longitudinal Study of Adult Health (ELSA-Brasil).

Assessing risk factors exposure, such as stress in the workplace during adulthood, may contribute to detecting early signs of cognitive impairment in order to implement effective actions to improve brain health and consequently to decrease cognitive disorders later in life. In this cross-sectional study, we aimed to investigate whether work-related stress is associated with low cognitive performance in middle-aged adults from the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) and whether social support mediates this relationship. Work-related stress was evaluated in 9,969 workers using the Demand-Control-Support Questionnaire. Cognitive function was assessed using the verbal fluency, trail-making version B, and delayed recall word tests. Work-related stress was associated with lower performance on the delayed recall, verbal fluency, and executive function tests in middle-aged adults. Social support may mediate the association between work demands and cognitive performance. These findings support that work-related stress is associated with cognitive performance during adulthood.

Work-related stress and cognitive performance among middle-aged adults: The Brazilian Longitudinal Study of Adult Health (ELSA-Brasil). Assessing risk factors exposure, such as stress in the workplace during adulthood, may contribute to detecting early signs of cognitive impairment in order to implement effective actions to improve brain health and consequently to decrease cognitive disorders later in life. In this cross-sectional study, we aimed to investigate whether work-related stress is associated with low cognitive performance in middle-aged adults from the Brazilian Longitudinal Study of Adult Health (ELSA-Brasil) and whether social support mediates this relationship. Work-related stress was evaluated in 9,969 workers using the Demand-Control-Support Questionnaire. Cognitive function was assessed using the verbal fluency, trail-making version B, and delayed recall word tests. Work-related stress was associated with lower performance on the delayed recall, verbal fluency, and executive function tests in middle-aged adults. Social support may mediate the association between work demands and cognitive performance. These findings support that work-related stress is associated with cognitive performance during adulthood.

pubmed23n0567_20779

Episodic death across species of desert shrubs.

Extreme events shape population and community trajectories. We report episodic mortality across common species of thousands of long-lived perennials individually tagged and monitored for 20 years in the Colorado Desert of California following severe regional drought. Demographic records from 1984 to 2004 show 15 years of virtual stasis in populations of adult shrubs and cacti, punctuated by a 55-100% die-off of six of the seven most common perennial species. In this episode, adults that experienced reduced growth in a lesser drought during 1984-1989 failed to survive the drought of 2002. The significance of this event is potentially profound because population dynamics of long-lived plants can be far more strongly affected by deaths of adults, which in deserts potentially live for centuries, than by seedling births or deaths. Differential mortality and rates of recovery during and after extreme climatic events quite likely determine the species composition of plant and associated animal communities for at least decades. The die-off recorded in this closely monitored community provides a unique window into the mechanics of this process of species decline and replacement.

Episodic death across species of desert shrubs. Extreme events shape population and community trajectories. We report episodic mortality across common species of thousands of long-lived perennials individually tagged and monitored for 20 years in the Colorado Desert of California following severe regional drought. Demographic records from 1984 to 2004 show 15 years of virtual stasis in populations of adult shrubs and cacti, punctuated by a 55-100% die-off of six of the seven most common perennial species. In this episode, adults that experienced reduced growth in a lesser drought during 1984-1989 failed to survive the drought of 2002. The significance of this event is potentially profound because population dynamics of long-lived plants can be far more strongly affected by deaths of adults, which in deserts potentially live for centuries, than by seedling births or deaths. Differential mortality and rates of recovery during and after extreme climatic events quite likely determine the species composition of plant and associated animal communities for at least decades. The die-off recorded in this closely monitored community provides a unique window into the mechanics of this process of species decline and replacement.

pubmed23n0762_8193

Testicular phosphoproteome in perfluorododecanoic acid-exposed rats.

Perfluorododecanoic acid (PFDoA) is a common environmental pollutant, which has been detected in human sera and has adverse effects on testicular function in animal models. Exploring phosphorylation events in testes helps elucidate the specific phosphorylation signals involved in testicular toxicity of PFDoA. Combining efficient prefractionation of tryptic peptide mixtures using self-packed reversed phase C18 columns with TiO2 and IMAC phosphopeptide enrichment techniques followed by 2D-LC-MS/MS, we identified 4077 unique phosphopeptides from 1777 proteins with a false discovery rate below 1.0% in the testes of rats exposed to PFDoA for 110 days. In addition, 937 novel phosphorylation sites were discovered in testicular proteins. Hundreds of phosphorylated proteins identified might be involved in spermatogenesis and sperm function. With increasing PFDoA dosage, the number of casein kinase 2 kinase-modified peptides significantly increased. Pathway analysis suggested that the mitogen-activated protein kinase pathway and cell division cycle protein 2 (CDC2) may have contributed to sperm activity and testicular function. By in vitro experiments, CDC2 phosphorylation activity was found to be likely involved in PFDoA-induced toxicity in Leydig cells. This study provides the first examination of the whole proteins' phosphorylation profile in rat testis and suggests that the MAPK pathway and CDC2 protein phosphorylation are critical for PFDoA testicular toxicity.

Testicular phosphoproteome in perfluorododecanoic acid-exposed rats. Perfluorododecanoic acid (PFDoA) is a common environmental pollutant, which has been detected in human sera and has adverse effects on testicular function in animal models. Exploring phosphorylation events in testes helps elucidate the specific phosphorylation signals involved in testicular toxicity of PFDoA. Combining efficient prefractionation of tryptic peptide mixtures using self-packed reversed phase C18 columns with TiO2 and IMAC phosphopeptide enrichment techniques followed by 2D-LC-MS/MS, we identified 4077 unique phosphopeptides from 1777 proteins with a false discovery rate below 1.0% in the testes of rats exposed to PFDoA for 110 days. In addition, 937 novel phosphorylation sites were discovered in testicular proteins. Hundreds of phosphorylated proteins identified might be involved in spermatogenesis and sperm function. With increasing PFDoA dosage, the number of casein kinase 2 kinase-modified peptides significantly increased. Pathway analysis suggested that the mitogen-activated protein kinase pathway and cell division cycle protein 2 (CDC2) may have contributed to sperm activity and testicular function. By in vitro experiments, CDC2 phosphorylation activity was found to be likely involved in PFDoA-induced toxicity in Leydig cells. This study provides the first examination of the whole proteins' phosphorylation profile in rat testis and suggests that the MAPK pathway and CDC2 protein phosphorylation are critical for PFDoA testicular toxicity.

pubmed23n0942_2103

Quality of Life and Stigma in Epilepsy, Perspectives from Selected Regions of Asia and Sub-Saharan Africa.

Epilepsy is an important and common worldwide public health problem that affects people of all ages. A significant number of individuals with epilepsy will be intractable to medication. These individuals experience an elevated mortality rate and negative psychosocial consequences of recurrent seizures. Surgery of epilepsy is highly effective to stop seizures in well-selected individuals, and seizure freedom is the most desirable result of epilepsy treatment due to the positive improvements in psychosocial function and the elimination of excess mortality associated with intractable epilepsy. Globally, there is inadequate data to fully assess epilepsy-related quality of life and stigma, although the preponderance of information we have points to a significant negative impact on people with epilepsy (PWE) and families of PWE. This review of the psychosocial impact of epilepsy focuses on regions of Asia and Sub-Saharan Africa that have been analyzed with population study approaches to determine the prevalence of epilepsy, treatment gaps, as well as factors impacting psychosocial function of PWE and their families. This review additionally identifies models of care for medically intractable epilepsy that have potential to significantly improve psychosocial function.

Quality of Life and Stigma in Epilepsy, Perspectives from Selected Regions of Asia and Sub-Saharan Africa. Epilepsy is an important and common worldwide public health problem that affects people of all ages. A significant number of individuals with epilepsy will be intractable to medication. These individuals experience an elevated mortality rate and negative psychosocial consequences of recurrent seizures. Surgery of epilepsy is highly effective to stop seizures in well-selected individuals, and seizure freedom is the most desirable result of epilepsy treatment due to the positive improvements in psychosocial function and the elimination of excess mortality associated with intractable epilepsy. Globally, there is inadequate data to fully assess epilepsy-related quality of life and stigma, although the preponderance of information we have points to a significant negative impact on people with epilepsy (PWE) and families of PWE. This review of the psychosocial impact of epilepsy focuses on regions of Asia and Sub-Saharan Africa that have been analyzed with population study approaches to determine the prevalence of epilepsy, treatment gaps, as well as factors impacting psychosocial function of PWE and their families. This review additionally identifies models of care for medically intractable epilepsy that have potential to significantly improve psychosocial function.

pubmed23n0112_15360

Drowning mortality in Los Angeles County, 1976 to 1984.

Drowning is the fourth leading cause of unintentional injury death in Los Angeles County. We examined data collected by the Los Angeles County Coroner's Office on drownings that occurred in the county from 1976 through 1984. There were 1587 drownings (1130 males and 457 females) during this nine-year period, for an annual rate of 2.36 drownings per 100,000 persons (3.44 for males and 1.33 for females). The largest proportion of drownings (44.5%) for both sexes, and in almost every age group, occurred in private swimming pools. Children 2 to 3 years of age had the highest swimming-pool drowning rate (7.95). The elderly also experienced high drowning rates, primarily in swimming pools and bathtubs. Drowning-site profiles varied dramatically by age and sex. These findings indicate a need for Los Angeles County to address the problem of drownings among infants and toddlers in private swimming pools and to investigate the failure of regulations requiring fencing of swimming pools to prevent these deaths. These findings also suggest several potential opportunities for preventive intervention by physicians and demonstrate that health professionals cannot rely on national drowning-site profiles when developing local drowning prevention strategies.

Drowning mortality in Los Angeles County, 1976 to 1984. Drowning is the fourth leading cause of unintentional injury death in Los Angeles County. We examined data collected by the Los Angeles County Coroner's Office on drownings that occurred in the county from 1976 through 1984. There were 1587 drownings (1130 males and 457 females) during this nine-year period, for an annual rate of 2.36 drownings per 100,000 persons (3.44 for males and 1.33 for females). The largest proportion of drownings (44.5%) for both sexes, and in almost every age group, occurred in private swimming pools. Children 2 to 3 years of age had the highest swimming-pool drowning rate (7.95). The elderly also experienced high drowning rates, primarily in swimming pools and bathtubs. Drowning-site profiles varied dramatically by age and sex. These findings indicate a need for Los Angeles County to address the problem of drownings among infants and toddlers in private swimming pools and to investigate the failure of regulations requiring fencing of swimming pools to prevent these deaths. These findings also suggest several potential opportunities for preventive intervention by physicians and demonstrate that health professionals cannot rely on national drowning-site profiles when developing local drowning prevention strategies.

pubmed23n0103_6278

Ciprofloxacin, azlocillin, ceftizoxime and amikacin alone and in combination against gram-negative bacilli in an infected chamber model.

Ciprofloxacin, azlocillin, ceftizoxime, and amikacin were studied alone and in combination against six Enterobacteriaceae and six strains of Pseudomonas aeruginosa in an infected chamber model in rabbits simulating a closed space infection. In-vivo results were compared with in-vitro tests of inhibition, killing and synergy. Ciprofloxacin was the most effective single agent, with efficacy against five of the six Enterobacteriaceae when used in low doses, and two of the six pseudomonads when used in high doses. The development of in-vitro resistance to ciprofloxacin was observed during therapy in strains which failed to be eradicated. Ciprofloxacin and azlocillin together was the most effective regimen, with efficacy against eleven of the twelve strains. Synergy, as determined by chequerboard testing, did not correlate with in-vivo outcome. Unlike mezlocillin, azlocillin, ceftizoxime or amikacin, MIC testing of ciprofloxacin was predictive of in-vivo success.

Ciprofloxacin, azlocillin, ceftizoxime and amikacin alone and in combination against gram-negative bacilli in an infected chamber model. Ciprofloxacin, azlocillin, ceftizoxime, and amikacin were studied alone and in combination against six Enterobacteriaceae and six strains of Pseudomonas aeruginosa in an infected chamber model in rabbits simulating a closed space infection. In-vivo results were compared with in-vitro tests of inhibition, killing and synergy. Ciprofloxacin was the most effective single agent, with efficacy against five of the six Enterobacteriaceae when used in low doses, and two of the six pseudomonads when used in high doses. The development of in-vitro resistance to ciprofloxacin was observed during therapy in strains which failed to be eradicated. Ciprofloxacin and azlocillin together was the most effective regimen, with efficacy against eleven of the twelve strains. Synergy, as determined by chequerboard testing, did not correlate with in-vivo outcome. Unlike mezlocillin, azlocillin, ceftizoxime or amikacin, MIC testing of ciprofloxacin was predictive of in-vivo success.

pubmed23n0920_11547

Relationship of Adverse Family Experiences to Resilience and School Engagement Among Vermont Youth.

Adverse childhood experiences (ACEs) are associated with a range of health outcomes and risk behaviors. In 2011-2012, the National Survey of Children's Health (NSCH) included questions about adverse family experiences (AFEs). AFE survey questions are similar to ACE questions, except there are no questions about emotional/physical/sexual trauma, and questions are asked of parents rather than children. Although the relationship between ACEs and work/school absenteeism has been studied, the relationships between AFEs of school-aged children, school performance, and buffering behaviors have not been explored in depth. We examined AFEs and measures of resilience and school engagement among 1330 Vermont children (6-17 years) included in the NSCH, using descriptive, bivariate, and multivariable analyses. The most prevalent AFEs were divorce/separation of parents; family income hardship; substance use problems; and mental illness, suicidality, or severe depression. Adjusting for sex, age, special health care needs, poverty level, and maternal physical/mental-emotional health status, children who had three or more AFEs had lower odds of completing all required homework [adjusted odds ratio (AOR) 3.3, 95% confidence interval (CI) 1.7-6.3] and higher odds of failing to exhibit resilience (AOR 2.1, 95% CI 1.2-3.8), compared to children having no AFEs. Children with three or more AFEs had difficulty engaging in school and completing homework, though poor outcomes were buffered when children showed resilience. Parents, school-based mental health professionals, and teachers could help identify children who may be less resilient and have difficulties completing homework assignments. Preventive approaches to children's emotional problems (e.g., promoting family health, using family-based approaches to treat emotional/behavioral problems) could be applied in schools and communities to foster resilience and improve school engagement of children.

Relationship of Adverse Family Experiences to Resilience and School Engagement Among Vermont Youth. Adverse childhood experiences (ACEs) are associated with a range of health outcomes and risk behaviors. In 2011-2012, the National Survey of Children's Health (NSCH) included questions about adverse family experiences (AFEs). AFE survey questions are similar to ACE questions, except there are no questions about emotional/physical/sexual trauma, and questions are asked of parents rather than children. Although the relationship between ACEs and work/school absenteeism has been studied, the relationships between AFEs of school-aged children, school performance, and buffering behaviors have not been explored in depth. We examined AFEs and measures of resilience and school engagement among 1330 Vermont children (6-17 years) included in the NSCH, using descriptive, bivariate, and multivariable analyses. The most prevalent AFEs were divorce/separation of parents; family income hardship; substance use problems; and mental illness, suicidality, or severe depression. Adjusting for sex, age, special health care needs, poverty level, and maternal physical/mental-emotional health status, children who had three or more AFEs had lower odds of completing all required homework [adjusted odds ratio (AOR) 3.3, 95% confidence interval (CI) 1.7-6.3] and higher odds of failing to exhibit resilience (AOR 2.1, 95% CI 1.2-3.8), compared to children having no AFEs. Children with three or more AFEs had difficulty engaging in school and completing homework, though poor outcomes were buffered when children showed resilience. Parents, school-based mental health professionals, and teachers could help identify children who may be less resilient and have difficulties completing homework assignments. Preventive approaches to children's emotional problems (e.g., promoting family health, using family-based approaches to treat emotional/behavioral problems) could be applied in schools and communities to foster resilience and improve school engagement of children.

pubmed23n1079_8763

Mixed hierarchical local structure in a disordered metal-organic framework.

Amorphous metal-organic frameworks (MOFs) are an emerging class of materials. However, their structural characterisation represents a significant challenge. Fe-BTC, and the commercial equivalent Basolite® F300, are MOFs with incredibly diverse catalytic ability, yet their disordered structures remain poorly understood. Here, we use advanced electron microscopy to identify a nanocomposite structure of Fe-BTC where nanocrystalline domains are embedded within an amorphous matrix, whilst synchrotron total scattering measurements reveal the extent of local atomic order within Fe-BTC. We use a polymerisation-based algorithm to generate an atomistic structure for Fe-BTC, the first example of this methodology applied to the amorphous MOF field outside the well-studied zeolitic imidazolate framework family. This demonstrates the applicability of this computational approach towards the modelling of other amorphous MOF systems with potential generality towards all MOF chemistries and connectivities. We find that the structures of Fe-BTC and Basolite® F300 can be represented by models containing a mixture of short- and medium-range order with a greater proportion of medium-range order in Basolite® F300 than in Fe-BTC. We conclude by discussing how our approach may allow for high-throughput computational discovery of functional, amorphous MOFs.

Mixed hierarchical local structure in a disordered metal-organic framework. Amorphous metal-organic frameworks (MOFs) are an emerging class of materials. However, their structural characterisation represents a significant challenge. Fe-BTC, and the commercial equivalent Basolite® F300, are MOFs with incredibly diverse catalytic ability, yet their disordered structures remain poorly understood. Here, we use advanced electron microscopy to identify a nanocomposite structure of Fe-BTC where nanocrystalline domains are embedded within an amorphous matrix, whilst synchrotron total scattering measurements reveal the extent of local atomic order within Fe-BTC. We use a polymerisation-based algorithm to generate an atomistic structure for Fe-BTC, the first example of this methodology applied to the amorphous MOF field outside the well-studied zeolitic imidazolate framework family. This demonstrates the applicability of this computational approach towards the modelling of other amorphous MOF systems with potential generality towards all MOF chemistries and connectivities. We find that the structures of Fe-BTC and Basolite® F300 can be represented by models containing a mixture of short- and medium-range order with a greater proportion of medium-range order in Basolite® F300 than in Fe-BTC. We conclude by discussing how our approach may allow for high-throughput computational discovery of functional, amorphous MOFs.

pubmed23n0210_4703

The human growth hormone gene family: structure and evolution of the chromosomal locus.

The structure of the human growth hormone gene cluster has been determined over a 78 kilobase region of DNA by the study of two overlapping cosmids. There are two growth hormone genes interspersed with three chorionic somatomammotropin genes, all in the same transcriptional orientation. One of the growth hormone genes lies in an active chromatin conformation in the pituitary and at least one of the chorionic somatomammotropin genes lies in an active chromatin conformation in the placenta. The two groups of genes are highly homologous throughout their 5' flanking and coding sequences, but diverge in their 3' flanking regions which raises the paradox of how genes so similar in structural and flanking sequences can be so differentially regulated. Analysis of the sequences of the genes and identification of at least three different classes of duplication units interspersed throughout the five gene cluster suggests that the cluster evolved quite recently and that the mechanism of gene duplication involved homologous but unequal exchange between middle repetitive elements of the Alu family.

The human growth hormone gene family: structure and evolution of the chromosomal locus. The structure of the human growth hormone gene cluster has been determined over a 78 kilobase region of DNA by the study of two overlapping cosmids. There are two growth hormone genes interspersed with three chorionic somatomammotropin genes, all in the same transcriptional orientation. One of the growth hormone genes lies in an active chromatin conformation in the pituitary and at least one of the chorionic somatomammotropin genes lies in an active chromatin conformation in the placenta. The two groups of genes are highly homologous throughout their 5' flanking and coding sequences, but diverge in their 3' flanking regions which raises the paradox of how genes so similar in structural and flanking sequences can be so differentially regulated. Analysis of the sequences of the genes and identification of at least three different classes of duplication units interspersed throughout the five gene cluster suggests that the cluster evolved quite recently and that the mechanism of gene duplication involved homologous but unequal exchange between middle repetitive elements of the Alu family.

pubmed23n0099_20529

beta-Endorphin: surface binding and internalization in thymoma cells.

The opioid peptide beta-endorphin binds to specific nonopioid binding sites (Mr 72,000) that are present on the surface of thymoma cells. beta-Endorphin is then internalized, apparently via the Mr 72,000 species, and is subsequently found within intracellular, vesicular structures. This process is accompanied by the down-regulation of the Mr 72,000 binding sites. Our findings suggest that beta-endorphin may modulate cellular functions, such as T-lymphocyte proliferation, at intracellular rather than cell surface sites.

beta-Endorphin: surface binding and internalization in thymoma cells. The opioid peptide beta-endorphin binds to specific nonopioid binding sites (Mr 72,000) that are present on the surface of thymoma cells. beta-Endorphin is then internalized, apparently via the Mr 72,000 species, and is subsequently found within intracellular, vesicular structures. This process is accompanied by the down-regulation of the Mr 72,000 binding sites. Our findings suggest that beta-endorphin may modulate cellular functions, such as T-lymphocyte proliferation, at intracellular rather than cell surface sites.

pubmed23n1091_4541

Diagnosis of an adenocarcinoma of Müllerian origin made as a result of its metastasis in the mouth floor.

Metastases derived from primary tumours distant to the oral and maxillofacial region account for only 1% of all malignancies at that location, usually with a poor prognosis. In women, the primary tumours that most frequently metastasise to the oral cavity are breast adenocarcinomas affecting the jawbones and soft tissues (41% and 24%, respectively), followed by adrenal glands and female genital organs to the jawbones (8%), and female genital organs to the soft tissues (15%). Metastatic tumours of the mouth are a challenging diagnosis because of their exceptional occurrence. We report on the case of an 83-year-old woman who consulted for a bleeding ulcerated tumour on the floor of the mouth. A biopsy-confirmed metastasis of an adenocarcinoma of Müllerian origin. The oral lesion was the first sign of undetected cancer. The patient agreed to surgical resection and was further referred to palliative care for her symptoms.

Diagnosis of an adenocarcinoma of Müllerian origin made as a result of its metastasis in the mouth floor. Metastases derived from primary tumours distant to the oral and maxillofacial region account for only 1% of all malignancies at that location, usually with a poor prognosis. In women, the primary tumours that most frequently metastasise to the oral cavity are breast adenocarcinomas affecting the jawbones and soft tissues (41% and 24%, respectively), followed by adrenal glands and female genital organs to the jawbones (8%), and female genital organs to the soft tissues (15%). Metastatic tumours of the mouth are a challenging diagnosis because of their exceptional occurrence. We report on the case of an 83-year-old woman who consulted for a bleeding ulcerated tumour on the floor of the mouth. A biopsy-confirmed metastasis of an adenocarcinoma of Müllerian origin. The oral lesion was the first sign of undetected cancer. The patient agreed to surgical resection and was further referred to palliative care for her symptoms.

pubmed23n0570_9603

Disease activity in pregnant women with Crohn's disease and birth outcomes: a regional Danish cohort study.

CD is associated with increased risk of adverse birth outcomes, but existing studies have not assessed the impact of disease activity during pregnancy. We examined the impact of disease activity on birth outcomes: LBW, preterm birth, LBW at term, and CAs. All births by CD women in North Jutland County, Denmark, from January 1, 1977 to December 31, 2005, were evaluated in a cohort study based on linkage between the Danish National Registry of Patients and the Medical Birth Registry. After identification of all births by CD women, review of medical records allowed collection of clinical details (including disease activity and drug therapy during pregnancy). The exposed cohort (N = 71) constituted pregnancies with low/moderate-high disease activity during pregnancy, and the unexposed cohort (N = 86) those with inactive disease. Logistic regression analyses were used to estimate the adjusted relative risks (with 95% confidence intervals) for adverse birth outcomes associated with disease activity in CD pregnancies. In subanalysis, we examined the impact of moderate-high activity. In women with disease activity, the adjusted risks of LBW, LBW at term, preterm birth, and CAs were 0.2 (0.0-2.6), 0.4 (0.0-3.7), 2.4 (0.6-9.5), and 0.8 (0.2-3.8), respectively. The crude risk of preterm birth was 3.4 (1.1-10.6) in those with moderate-high disease activity. Disease activity during pregnancy only increased the risk of preterm birth (especially in those with high disease activity). Further research is needed to assess the critical impact of disease activity in larger cohorts of CD women.

Disease activity in pregnant women with Crohn's disease and birth outcomes: a regional Danish cohort study. CD is associated with increased risk of adverse birth outcomes, but existing studies have not assessed the impact of disease activity during pregnancy. We examined the impact of disease activity on birth outcomes: LBW, preterm birth, LBW at term, and CAs. All births by CD women in North Jutland County, Denmark, from January 1, 1977 to December 31, 2005, were evaluated in a cohort study based on linkage between the Danish National Registry of Patients and the Medical Birth Registry. After identification of all births by CD women, review of medical records allowed collection of clinical details (including disease activity and drug therapy during pregnancy). The exposed cohort (N = 71) constituted pregnancies with low/moderate-high disease activity during pregnancy, and the unexposed cohort (N = 86) those with inactive disease. Logistic regression analyses were used to estimate the adjusted relative risks (with 95% confidence intervals) for adverse birth outcomes associated with disease activity in CD pregnancies. In subanalysis, we examined the impact of moderate-high activity. In women with disease activity, the adjusted risks of LBW, LBW at term, preterm birth, and CAs were 0.2 (0.0-2.6), 0.4 (0.0-3.7), 2.4 (0.6-9.5), and 0.8 (0.2-3.8), respectively. The crude risk of preterm birth was 3.4 (1.1-10.6) in those with moderate-high disease activity. Disease activity during pregnancy only increased the risk of preterm birth (especially in those with high disease activity). Further research is needed to assess the critical impact of disease activity in larger cohorts of CD women.

pubmed23n0532_17477

Robotic radical prostatectomy with preservation of the prostatic fascia: a feasibility study.

To describe a feasibility study of our ability to preserve the prostatic fascia in men undergoing robotic radical prostatectomy. The prostate is covered anterolaterally by prostatic fascia, also called lateral pelvic fascia or the parietal layer of endopelvic fascia. The prostatic fascia is rich in vessels, nerves, and smooth muscle. We hypothesized that preservation of this fascial layer may result in improved postoperative potency. The technique was first attempted in 15 men undergoing radical cystoprostatectomy, in which accidental entry to the prostatic tissue is not critical. Thereafter, it was performed in 6 impotent men undergoing robotic radical prostatectomy. The fascia was excised and stained for prostate-specific antigen and neural and muscle tissue. The technique was then performed in 35 potent men (Sexual Health Inventory for Men score greater than 21) undergoing robotic radical prostatectomy. Postoperative potency was evaluated with a self-administered questionnaire (Sexual Health Inventory for Men). Under the magnification of the da Vinci robotic system, and also shown histologically, the prostatic fascia is a multifascial layer of fibrovascular tissue, covering the anterolateral aspect of the prostate. It stains positive for smooth muscle and nerves, but negative for prostate-specific antigen. The amount of neural tissue in the fascia is variable, but never exceeds that in the neurovascular bundle. At 12 months of follow-up, 34 (97%) of 35 men undergoing fascia-preserving robotic radical prostatectomy had erections strong enough for vaginal penetration, and 30 (86%) had normal erections (Sexual Health Inventory for Men greater than 21). Preservation of the prostatic fascia is safe and feasible, without compromising the surgical margins, and allows enhanced preservation of neural tissue during robotic prostatectomy with an apparent improvement in potency.

Robotic radical prostatectomy with preservation of the prostatic fascia: a feasibility study. To describe a feasibility study of our ability to preserve the prostatic fascia in men undergoing robotic radical prostatectomy. The prostate is covered anterolaterally by prostatic fascia, also called lateral pelvic fascia or the parietal layer of endopelvic fascia. The prostatic fascia is rich in vessels, nerves, and smooth muscle. We hypothesized that preservation of this fascial layer may result in improved postoperative potency. The technique was first attempted in 15 men undergoing radical cystoprostatectomy, in which accidental entry to the prostatic tissue is not critical. Thereafter, it was performed in 6 impotent men undergoing robotic radical prostatectomy. The fascia was excised and stained for prostate-specific antigen and neural and muscle tissue. The technique was then performed in 35 potent men (Sexual Health Inventory for Men score greater than 21) undergoing robotic radical prostatectomy. Postoperative potency was evaluated with a self-administered questionnaire (Sexual Health Inventory for Men). Under the magnification of the da Vinci robotic system, and also shown histologically, the prostatic fascia is a multifascial layer of fibrovascular tissue, covering the anterolateral aspect of the prostate. It stains positive for smooth muscle and nerves, but negative for prostate-specific antigen. The amount of neural tissue in the fascia is variable, but never exceeds that in the neurovascular bundle. At 12 months of follow-up, 34 (97%) of 35 men undergoing fascia-preserving robotic radical prostatectomy had erections strong enough for vaginal penetration, and 30 (86%) had normal erections (Sexual Health Inventory for Men greater than 21). Preservation of the prostatic fascia is safe and feasible, without compromising the surgical margins, and allows enhanced preservation of neural tissue during robotic prostatectomy with an apparent improvement in potency.

pubmed23n0948_11589

Frequency of abnormal fractional flow reserve measurements among major coronary arteries.

Fractional flow reserve(FFR) is a validated tool for evaluating functional severity and guiding the revascularization of angiographically moderate coronary artery lesions. To study if there is a higher frequency of positive FFR measurements in the left anterior descending(LAD) versus other major coronary arteries and also evaluate the differences in the total length of the stent placed. A retrospective cohort study including all subjects (January 2011 to December 2015) who had fractional flow reserve (FFR) measured during coronary catheterization was conducted. Coronary catheterizations with FFR at a single tertiary care center were reviewed and FFR ≤ 0.80 post adenosine was deemed positive. The differences in the baseline characteristics and the degree of stenosis were compared between the different vessel groups. Of the 758 vessels included in the analysis, the majority were LAD(51.3%) followed by right coronary artery(RCA)(22.8%), Circumflex(22.2%), Left main(2.2%), and Ramus intermedius(1.5%). 25.1% of 758 vessels were FFR positive. The proportion of positive FFR were higher among LAD versus other vessels(33.2%vs.16.5%,p &lt; 0.001), while no differences were noted between RCA and circumflex(p = 0.87) or other vessels excluding LAD(p = 0.69). Of 175 patients who received stents, no statistical difference was noted in the median[range] total length of the stent between LAD(22[9-64]) and the other coronary arteries (18[8-42])(p = 0.19). In patients with an FFR &lt;0.75, we found that the stent length(median [range]) was significantly longer in LAD(28[9-42]) than the other coronary arteries(18[8-42])(p = 0.03). In our study, FFR was almost twice as likely to be positive in the LAD when compared to other major coronary arteries. Furthermore, there was a trend towards FFR positive LAD lesions needing longer stents than other coronary arteries. This data should encourage operators to evaluate moderate, long lesions in the LAD with FFR, as they have a higher probability of functional significance.

Frequency of abnormal fractional flow reserve measurements among major coronary arteries. Fractional flow reserve(FFR) is a validated tool for evaluating functional severity and guiding the revascularization of angiographically moderate coronary artery lesions. To study if there is a higher frequency of positive FFR measurements in the left anterior descending(LAD) versus other major coronary arteries and also evaluate the differences in the total length of the stent placed. A retrospective cohort study including all subjects (January 2011 to December 2015) who had fractional flow reserve (FFR) measured during coronary catheterization was conducted. Coronary catheterizations with FFR at a single tertiary care center were reviewed and FFR ≤ 0.80 post adenosine was deemed positive. The differences in the baseline characteristics and the degree of stenosis were compared between the different vessel groups. Of the 758 vessels included in the analysis, the majority were LAD(51.3%) followed by right coronary artery(RCA)(22.8%), Circumflex(22.2%), Left main(2.2%), and Ramus intermedius(1.5%). 25.1% of 758 vessels were FFR positive. The proportion of positive FFR were higher among LAD versus other vessels(33.2%vs.16.5%,p &lt; 0.001), while no differences were noted between RCA and circumflex(p = 0.87) or other vessels excluding LAD(p = 0.69). Of 175 patients who received stents, no statistical difference was noted in the median[range] total length of the stent between LAD(22[9-64]) and the other coronary arteries (18[8-42])(p = 0.19). In patients with an FFR &lt;0.75, we found that the stent length(median [range]) was significantly longer in LAD(28[9-42]) than the other coronary arteries(18[8-42])(p = 0.03). In our study, FFR was almost twice as likely to be positive in the LAD when compared to other major coronary arteries. Furthermore, there was a trend towards FFR positive LAD lesions needing longer stents than other coronary arteries. This data should encourage operators to evaluate moderate, long lesions in the LAD with FFR, as they have a higher probability of functional significance.

pubmed23n1141_22455

A prospective cohort study of methotrexate plus idarubicin in newly diagnosed primary CNS lymphoma.

High-dose methotrexate (HD-MTX)-based chemotherapy regimen is the first-line option for primary central nervous system lymphoma (PCNSL). This prospective cohort study aimed to evaluate the efficacy and adverse effects of HD-MTX plus idarubicin (IDA) in patients with newly diagnosed immunocompetent PCNSL. We recruited newly diagnosed PCNSL patients from January 2017 to August 2020. Patients were assigned into two groups: HD-MTX monotherapy and HD-MTX plus IDA (HD-MTX/IDA). In the HD-MTX monotherapy group, patients were treated with MTX 8 g/m<sup2</sup alone on day 1, while the HD-MTX/IDA group received MTX 8 g/m<sup2</sup on day 1 and IDA 10 mg/m<sup2</sup on day 2. Treatments were repeated every 3 weeks for 8 cycles except for progression and/or unacceptable toxicity. We recruited 61 PCNSL patients, including 36 in the HD-MTX and 25 in the HD-MTX/IDA group. The CR rate was 68% in the HD-MTX/IDA group and 72.22% of patients in the HD-MTX monotherapy group (p = 0.7221), while the overall response rate was 72% vs. 77.78% (p = 0.6063). Median PFS in HD-MTX/IDA group and HD-MTX monotherapy group were 15.6 months and 18.5 months, respectively (p = 0.6374). Median OS was not reached in both groups. There were no significant differences in adverse effects between the two groups. The combination of IDA with HD-MTX showed no obvious therapeutic advantage over HD-MTX monotherapy in newly diagnosed patients with PCNSL. HD-MTX dose of 8 g/m<sup2</sup monotherapy can still provide better therapeutic benefits in patients with acceptable adverse effects. Future studies could explore HD-MTX in combination with other chemotherapeutic agents in the first-line treatment of PCNSL.

A prospective cohort study of methotrexate plus idarubicin in newly diagnosed primary CNS lymphoma. High-dose methotrexate (HD-MTX)-based chemotherapy regimen is the first-line option for primary central nervous system lymphoma (PCNSL). This prospective cohort study aimed to evaluate the efficacy and adverse effects of HD-MTX plus idarubicin (IDA) in patients with newly diagnosed immunocompetent PCNSL. We recruited newly diagnosed PCNSL patients from January 2017 to August 2020. Patients were assigned into two groups: HD-MTX monotherapy and HD-MTX plus IDA (HD-MTX/IDA). In the HD-MTX monotherapy group, patients were treated with MTX 8 g/m<sup2</sup alone on day 1, while the HD-MTX/IDA group received MTX 8 g/m<sup2</sup on day 1 and IDA 10 mg/m<sup2</sup on day 2. Treatments were repeated every 3 weeks for 8 cycles except for progression and/or unacceptable toxicity. We recruited 61 PCNSL patients, including 36 in the HD-MTX and 25 in the HD-MTX/IDA group. The CR rate was 68% in the HD-MTX/IDA group and 72.22% of patients in the HD-MTX monotherapy group (p = 0.7221), while the overall response rate was 72% vs. 77.78% (p = 0.6063). Median PFS in HD-MTX/IDA group and HD-MTX monotherapy group were 15.6 months and 18.5 months, respectively (p = 0.6374). Median OS was not reached in both groups. There were no significant differences in adverse effects between the two groups. The combination of IDA with HD-MTX showed no obvious therapeutic advantage over HD-MTX monotherapy in newly diagnosed patients with PCNSL. HD-MTX dose of 8 g/m<sup2</sup monotherapy can still provide better therapeutic benefits in patients with acceptable adverse effects. Future studies could explore HD-MTX in combination with other chemotherapeutic agents in the first-line treatment of PCNSL.

pubmed23n0742_20286

Mouse models for filovirus infections.

The filoviruses marburg- and ebolaviruses can cause severe hemorrhagic fever (HF) in humans and nonhuman primates. Because many cases have occurred in geographical areas lacking a medical research infrastructure, most studies of the pathogenesis of filoviral HF, and all efforts to develop drugs and vaccines, have been carried out in biocontainment laboratories in non-endemic countries, using nonhuman primates (NHPs), guinea pigs and mice as animal models. NHPs appear to closely mirror filoviral HF in humans (based on limited clinical data), but only small numbers may be used in carefully regulated experiments; much research is therefore done in rodents. Because of their availability in large numbers and the existence of a wealth of reagents for biochemical and immunological testing, mice have become the preferred small animal model for filovirus research. Since the first experiments following the initial 1967 marburgvirus outbreak, wild-type or mouse-adapted viruses have been tested in immunocompetent or immunodeficient mice. In this paper, we review how these types of studies have been used to investigate the pathogenesis of filoviral disease, identify immune responses to infection and evaluate antiviral drugs and vaccines. We also discuss the strengths and weaknesses of murine models for filovirus research, and identify important questions for further study.

Mouse models for filovirus infections. The filoviruses marburg- and ebolaviruses can cause severe hemorrhagic fever (HF) in humans and nonhuman primates. Because many cases have occurred in geographical areas lacking a medical research infrastructure, most studies of the pathogenesis of filoviral HF, and all efforts to develop drugs and vaccines, have been carried out in biocontainment laboratories in non-endemic countries, using nonhuman primates (NHPs), guinea pigs and mice as animal models. NHPs appear to closely mirror filoviral HF in humans (based on limited clinical data), but only small numbers may be used in carefully regulated experiments; much research is therefore done in rodents. Because of their availability in large numbers and the existence of a wealth of reagents for biochemical and immunological testing, mice have become the preferred small animal model for filovirus research. Since the first experiments following the initial 1967 marburgvirus outbreak, wild-type or mouse-adapted viruses have been tested in immunocompetent or immunodeficient mice. In this paper, we review how these types of studies have been used to investigate the pathogenesis of filoviral disease, identify immune responses to infection and evaluate antiviral drugs and vaccines. We also discuss the strengths and weaknesses of murine models for filovirus research, and identify important questions for further study.

pubmed23n1099_3044

GPU-accelerated multitiered iterative phasing algorithm for fluctuation X-ray scattering.

The multitiered iterative phasing (MTIP) algorithm is used to determine the biological structures of macromolecules from fluctuation scattering data. It is an iterative algorithm that reconstructs the electron density of the sample by matching the computed fluctuation X-ray scattering data to the external observations, and by simultaneously enforcing constraints in real and Fourier space. This paper presents the first ever MTIP algorithm acceleration efforts on contemporary graphics processing units (GPUs). The Compute Unified Device Architecture (CUDA) programming model is used to accelerate the MTIP algorithm on NVIDIA GPUs. The computational performance of the CUDA-based MTIP algorithm implementation outperforms the CPU-based version by an order of magnitude. Furthermore, the Heterogeneous-Compute Interface for Portability (HIP) runtime APIs are used to demonstrate portability by accelerating the MTIP algorithm across NVIDIA and AMD GPUs.

GPU-accelerated multitiered iterative phasing algorithm for fluctuation X-ray scattering. The multitiered iterative phasing (MTIP) algorithm is used to determine the biological structures of macromolecules from fluctuation scattering data. It is an iterative algorithm that reconstructs the electron density of the sample by matching the computed fluctuation X-ray scattering data to the external observations, and by simultaneously enforcing constraints in real and Fourier space. This paper presents the first ever MTIP algorithm acceleration efforts on contemporary graphics processing units (GPUs). The Compute Unified Device Architecture (CUDA) programming model is used to accelerate the MTIP algorithm on NVIDIA GPUs. The computational performance of the CUDA-based MTIP algorithm implementation outperforms the CPU-based version by an order of magnitude. Furthermore, the Heterogeneous-Compute Interface for Portability (HIP) runtime APIs are used to demonstrate portability by accelerating the MTIP algorithm across NVIDIA and AMD GPUs.

pubmed23n0664_18620

Online monitoring of particle mass flow rate in bottom spray fluid bed coating--development and application.

The primary purpose of this study is to develop a visiometric process analyzer for online monitoring of particle mass flow rate in the bottom spray fluid bed coating process. The secondary purpose is to investigate the influences of partition gap and air accelerator insert size on particle mass flow rate using the developed visiometric process analyzer. Particle movement in the region between the product chamber and partition column was captured using a high speed camera. Mean particle velocity and number of particles in the images were determined by particle image velocimetry and morphological image processing method respectively. Mass flow rate was calculated using particle velocity, number of particles in the images, particle density and size information. Particle velocity and number findings were validated using image tracking and manual particle counting techniques respectively. Validation experiments showed that the proposed method was accurate. Partition gap was found to influence particle mass flow rate by limiting the rate of solids flux into the partition column; the air accelerator insert was found to influence particle mass flow rate by a Venturi effect. Partition gap and air accelerator insert diameter needed to be adjusted accordingly in relation to the other variability sources and diameter of coating cores respectively. The potential, challenges and possible solutions of the proposed visiometric process analyzer were further discussed.

Online monitoring of particle mass flow rate in bottom spray fluid bed coating--development and application. The primary purpose of this study is to develop a visiometric process analyzer for online monitoring of particle mass flow rate in the bottom spray fluid bed coating process. The secondary purpose is to investigate the influences of partition gap and air accelerator insert size on particle mass flow rate using the developed visiometric process analyzer. Particle movement in the region between the product chamber and partition column was captured using a high speed camera. Mean particle velocity and number of particles in the images were determined by particle image velocimetry and morphological image processing method respectively. Mass flow rate was calculated using particle velocity, number of particles in the images, particle density and size information. Particle velocity and number findings were validated using image tracking and manual particle counting techniques respectively. Validation experiments showed that the proposed method was accurate. Partition gap was found to influence particle mass flow rate by limiting the rate of solids flux into the partition column; the air accelerator insert was found to influence particle mass flow rate by a Venturi effect. Partition gap and air accelerator insert diameter needed to be adjusted accordingly in relation to the other variability sources and diameter of coating cores respectively. The potential, challenges and possible solutions of the proposed visiometric process analyzer were further discussed.

pubmed23n0947_6479

Using Google Earth to Assess Shade for Sun Protection in Urban Recreation Spaces: Methods and Results.

Shade in public spaces can lower the risk of and sun burning and skin cancer. However, existing methods of auditing shade require travel between sites, and sunny weather conditions. This study aimed to evaluate the feasibility of free computer software-Google Earth-for assessing shade in urban open spaces. A shade projection method was developed that uses Google Earth street view and aerial images to estimate shade at solar noon on the summer solstice, irrespective of the date of image capture. Three researchers used the method to separately estimate shade cover over pre-defined activity areas in a sample of 45 New Zealand urban open spaces, including 24 playgrounds, 12 beaches and 9 outdoor pools. Outcome measures included method accuracy (assessed by comparison with a subsample of field observations of 10 of the settings) and inter-rater reliability. Of the 164 activity areas identified in the 45 settings, most (83%) had no shade cover. The method identified most activity areas in playgrounds (85%) and beaches (93%) and was accurate for assessing shade over these areas (predictive values of 100%). Only 8% of activity areas at outdoor pools were identified, due to a lack of street view images. Reliability for shade cover estimates was excellent (intraclass correlation coefficient of 0.97, 95% CI 0.97-0.98). Google Earth appears to be a reasonably accurate and reliable and shade audit tool for playgrounds and beaches. The findings are relevant for programmes focused on supporting the development of healthy urban open spaces.

Using Google Earth to Assess Shade for Sun Protection in Urban Recreation Spaces: Methods and Results. Shade in public spaces can lower the risk of and sun burning and skin cancer. However, existing methods of auditing shade require travel between sites, and sunny weather conditions. This study aimed to evaluate the feasibility of free computer software-Google Earth-for assessing shade in urban open spaces. A shade projection method was developed that uses Google Earth street view and aerial images to estimate shade at solar noon on the summer solstice, irrespective of the date of image capture. Three researchers used the method to separately estimate shade cover over pre-defined activity areas in a sample of 45 New Zealand urban open spaces, including 24 playgrounds, 12 beaches and 9 outdoor pools. Outcome measures included method accuracy (assessed by comparison with a subsample of field observations of 10 of the settings) and inter-rater reliability. Of the 164 activity areas identified in the 45 settings, most (83%) had no shade cover. The method identified most activity areas in playgrounds (85%) and beaches (93%) and was accurate for assessing shade over these areas (predictive values of 100%). Only 8% of activity areas at outdoor pools were identified, due to a lack of street view images. Reliability for shade cover estimates was excellent (intraclass correlation coefficient of 0.97, 95% CI 0.97-0.98). Google Earth appears to be a reasonably accurate and reliable and shade audit tool for playgrounds and beaches. The findings are relevant for programmes focused on supporting the development of healthy urban open spaces.

pubmed23n1047_2319

The NK cell granule protein NKG7 regulates cytotoxic granule exocytosis and inflammation.

Immune-modulating therapies have revolutionized the treatment of chronic diseases, particularly cancer. However, their success is restricted and there is a need to identify new therapeutic targets. Here, we show that natural killer cell granule protein 7 (NKG7) is a regulator of lymphocyte granule exocytosis and downstream inflammation in a broad range of diseases. NKG7 expressed by CD4<sup+</sup and CD8<sup+</sup T cells played key roles in promoting inflammation during visceral leishmaniasis and malaria-two important parasitic diseases. Additionally, NKG7 expressed by natural killer cells was critical for controlling cancer initiation, growth and metastasis. NKG7 function in natural killer and CD8<sup+</sup T cells was linked with their ability to regulate the translocation of CD107a to the cell surface and kill cellular targets, while NKG7 also had a major impact on CD4<sup+</sup T cell activation following infection. Thus, we report a novel therapeutic target expressed on a range of immune cells with functions in different immune responses.

The NK cell granule protein NKG7 regulates cytotoxic granule exocytosis and inflammation. Immune-modulating therapies have revolutionized the treatment of chronic diseases, particularly cancer. However, their success is restricted and there is a need to identify new therapeutic targets. Here, we show that natural killer cell granule protein 7 (NKG7) is a regulator of lymphocyte granule exocytosis and downstream inflammation in a broad range of diseases. NKG7 expressed by CD4<sup+</sup and CD8<sup+</sup T cells played key roles in promoting inflammation during visceral leishmaniasis and malaria-two important parasitic diseases. Additionally, NKG7 expressed by natural killer cells was critical for controlling cancer initiation, growth and metastasis. NKG7 function in natural killer and CD8<sup+</sup T cells was linked with their ability to regulate the translocation of CD107a to the cell surface and kill cellular targets, while NKG7 also had a major impact on CD4<sup+</sup T cell activation following infection. Thus, we report a novel therapeutic target expressed on a range of immune cells with functions in different immune responses.

pubmed23n0391_22847

Galectin-1 is overexpressed in nasal polyps under budesonide and inhibits eosinophil migration.

Because of the importance of galectins for various cellular activities, the influence of the glucocorticoid budesonide on the level of expression of galectins-1 and -3 was investigated in human nasal polyposis. Ten nasal polyps obtained from surgical resection were maintained for 24 hours in the presence of various concentrations of budesonide. As quantitatively demonstrated by means of computer-assisted microscopy, 250 ng/ml (the highest dose tested) induced a pronounced increase of galectin-1 expression. This feature was observed in nasal polyps from allergic patients but not in those from nonallergic patients. Since eosinophils represent the main inflammatory cell population in nasal polyps, we investigated the effect of galectin-1 on their migration levels by means of quantitative phase-contrast computer-assisted videomicroscopy. Our results show that galectin-1 (coated on plastic supports) markedly reduced the migration levels of eosinophils in comparison to P-selectin. On the cellular level, marked modifications in the polymerization/depolymerization dynamics of the actin cytoskeleton (as revealed by means of computer-assisted fluorescence microscopy) and, to a much lesser extent, an increase in the adhesiveness of eosinophils to tested substrata were detectable. The present study therefore reveals a new galectin-1-mediated mechanism of action for glucocorticoid-mediated anti-inflammatory effects.

Galectin-1 is overexpressed in nasal polyps under budesonide and inhibits eosinophil migration. Because of the importance of galectins for various cellular activities, the influence of the glucocorticoid budesonide on the level of expression of galectins-1 and -3 was investigated in human nasal polyposis. Ten nasal polyps obtained from surgical resection were maintained for 24 hours in the presence of various concentrations of budesonide. As quantitatively demonstrated by means of computer-assisted microscopy, 250 ng/ml (the highest dose tested) induced a pronounced increase of galectin-1 expression. This feature was observed in nasal polyps from allergic patients but not in those from nonallergic patients. Since eosinophils represent the main inflammatory cell population in nasal polyps, we investigated the effect of galectin-1 on their migration levels by means of quantitative phase-contrast computer-assisted videomicroscopy. Our results show that galectin-1 (coated on plastic supports) markedly reduced the migration levels of eosinophils in comparison to P-selectin. On the cellular level, marked modifications in the polymerization/depolymerization dynamics of the actin cytoskeleton (as revealed by means of computer-assisted fluorescence microscopy) and, to a much lesser extent, an increase in the adhesiveness of eosinophils to tested substrata were detectable. The present study therefore reveals a new galectin-1-mediated mechanism of action for glucocorticoid-mediated anti-inflammatory effects.

pubmed23n0925_9189

The Future of Surveillance in the Context of Cancer Predisposition: Through the Murky Looking Glass.

At least 10% of children with cancer harbor a disease-associated pathogenic variant in a known cancer predisposition gene. It is widely accepted that pathogenic variants affecting other genes, epigenetic factors, or abnormalities in additional gene products may contribute to the etiology of many more childhood cancers. Effective preventive measures exist for only a few cancer types associated with predisposing conditions, but the development and implementation of surveillance protocols aimed at reducing morbidity and mortality in at-risk children through the early detection of cancer has emerged as an important clinical tool. The articles in this <iClinical Cancer Research</i series present international consensus generated recommendations for surveillance for a wide spectrum of cancer predisposition syndromes affecting children. In this article, we explore the challenges and opportunities for researchers and practitioners in the many fields affiliated with pediatric cancer, and we offer insights into what the future might hold as we continue our efforts to mitigate the impact of cancer susceptibility on children, their families and society. <iClin Cancer Res; 23(21); e133-e7. ©2017 AACR</i<bSee all articles in the online-only <iCCR</i Pediatric Oncology Series.</b

The Future of Surveillance in the Context of Cancer Predisposition: Through the Murky Looking Glass. At least 10% of children with cancer harbor a disease-associated pathogenic variant in a known cancer predisposition gene. It is widely accepted that pathogenic variants affecting other genes, epigenetic factors, or abnormalities in additional gene products may contribute to the etiology of many more childhood cancers. Effective preventive measures exist for only a few cancer types associated with predisposing conditions, but the development and implementation of surveillance protocols aimed at reducing morbidity and mortality in at-risk children through the early detection of cancer has emerged as an important clinical tool. The articles in this <iClinical Cancer Research</i series present international consensus generated recommendations for surveillance for a wide spectrum of cancer predisposition syndromes affecting children. In this article, we explore the challenges and opportunities for researchers and practitioners in the many fields affiliated with pediatric cancer, and we offer insights into what the future might hold as we continue our efforts to mitigate the impact of cancer susceptibility on children, their families and society. <iClin Cancer Res; 23(21); e133-e7. ©2017 AACR</i<bSee all articles in the online-only <iCCR</i Pediatric Oncology Series.</b

pubmed23n0695_6138

Preparation of monodisperse poly(N-isopropylacrylamide) microgel particles with homogenous cross-link density distribution.

Monodisperse microgel latex with homogeneous cross-link density distribution within the particles was prepared by feeding the monomer and cross-linker into the reaction mixture in a regulated way during the polymerization. To determine the appropriate monomer feeding parameters, the kinetics of the particle formation was investigated by HPLC. The swelling and optical characteristics of the prepared homogenously cross-linked microgel particles were compared to the properties of inhomogenously cross-linked microgels prepared by the normal precipitation polymerization method. The distribution of the cross-link density within the particles inserts a great influence on the characteristics of the system. The degree of swelling of the homogeneous particles is significantly higher than that of the heterogeneous microgel particles. Furthermore, at room temperature the pNIPAm latex containing the homogeneously cross-linked particles is transparent, while the heterogeneously cross-linked particles form a highly turbid system at the same 0.1 wt% concentration.

Preparation of monodisperse poly(N-isopropylacrylamide) microgel particles with homogenous cross-link density distribution. Monodisperse microgel latex with homogeneous cross-link density distribution within the particles was prepared by feeding the monomer and cross-linker into the reaction mixture in a regulated way during the polymerization. To determine the appropriate monomer feeding parameters, the kinetics of the particle formation was investigated by HPLC. The swelling and optical characteristics of the prepared homogenously cross-linked microgel particles were compared to the properties of inhomogenously cross-linked microgels prepared by the normal precipitation polymerization method. The distribution of the cross-link density within the particles inserts a great influence on the characteristics of the system. The degree of swelling of the homogeneous particles is significantly higher than that of the heterogeneous microgel particles. Furthermore, at room temperature the pNIPAm latex containing the homogeneously cross-linked particles is transparent, while the heterogeneously cross-linked particles form a highly turbid system at the same 0.1 wt% concentration.

pubmed23n0786_3121

The effect of mortality salience on implicit bias.

Previous research in terror management theory has shown that when individuals are reminded of their mortality, negative evaluations of out-group members increase. This previous research has used a variety of methods to investigate the change in attitudes toward out-group members. These methods generally permit participants time to consciously reflect during responding In the present study, the authors hypothesized that reminding individuals of their mortality would increase implicit bias toward out-group members when measured with the Implicit Association Test (IAT). In the experiment, White participants were randomly assigned to complete a written essay about imagining their own death (i.e., mortality salience condition) or an essay regarding an upcoming exam (i.e., control condition), and then participants completed an evaluative IAT. The results revealed that participants who had been reminded of their mortality took longer to associate names of Black individuals with positive words when compared to a control group. The implications of these findings are considered.

The effect of mortality salience on implicit bias. Previous research in terror management theory has shown that when individuals are reminded of their mortality, negative evaluations of out-group members increase. This previous research has used a variety of methods to investigate the change in attitudes toward out-group members. These methods generally permit participants time to consciously reflect during responding In the present study, the authors hypothesized that reminding individuals of their mortality would increase implicit bias toward out-group members when measured with the Implicit Association Test (IAT). In the experiment, White participants were randomly assigned to complete a written essay about imagining their own death (i.e., mortality salience condition) or an essay regarding an upcoming exam (i.e., control condition), and then participants completed an evaluative IAT. The results revealed that participants who had been reminded of their mortality took longer to associate names of Black individuals with positive words when compared to a control group. The implications of these findings are considered.

pubmed23n0648_3076

Randomized trial of effect of bifocal and prismatic bifocal spectacles on myopic progression: two-year results.

To determine whether bifocal and prismatic bifocal spectacles could control myopia in children with high rates of myopic progression. This was a randomized controlled clinical trial. One hundred thirty-five (73 girls and 62 boys) myopic Chinese Canadian children (myopia of &gt; or =1.00 diopters [D]) with myopic progression of at least 0.50 D in the preceding year were randomly assigned to 1 of 3 treatments: (1) single-vision lenses (n = 41), (2) +1.50-D executive bifocals (n = 48), or (3) +1.50-D executive bifocals with a 3-prism diopters base-in prism in the near segment of each lens (n = 46). Myopic progression measured by an automated refractor under cycloplegia and increase in axial length (secondary) measured by ultrasonography at 6-month intervals for 24 months. Only the data of the right eye were used. Of the 135 children (mean age, 10.29 years [SE, 0.15 years]; mean visual acuity, -3.08 D [SE, 0.10 D]), 131 (97%) completed the trial after 24 months. Myopic progression averaged -1.55 D (SE, 0.12 D) for those who wore single-vision lenses, -0.96 D (SE, 0.09 D) for those who wore bifocals, and -0.70 D (SE, 0.10 D) for those who wore prismatic bifocals. Axial length increased an average of 0.62 mm (SE, 0.04 mm), 0.41 mm (SE, 0.04 mm), and 0.41 mm (SE, 0.05 mm), respectively. The treatment effect of bifocals (0.59 D) and prismatic bifocals (0.85 D) was significant (P &lt; .001) and both bifocal groups had less axial elongation (0.21 mm) than the single-vision lens group (P &lt; .001). Bifocal lenses can moderately slow myopic progression in children with high rates of progression after 24 months. APPLICATIONS TO CLINICAL PRACTICE: Bifocal spectacles may be considered for slowing myopic progression in children with an annual progression rate of at least 0.50 D.

Randomized trial of effect of bifocal and prismatic bifocal spectacles on myopic progression: two-year results. To determine whether bifocal and prismatic bifocal spectacles could control myopia in children with high rates of myopic progression. This was a randomized controlled clinical trial. One hundred thirty-five (73 girls and 62 boys) myopic Chinese Canadian children (myopia of &gt; or =1.00 diopters [D]) with myopic progression of at least 0.50 D in the preceding year were randomly assigned to 1 of 3 treatments: (1) single-vision lenses (n = 41), (2) +1.50-D executive bifocals (n = 48), or (3) +1.50-D executive bifocals with a 3-prism diopters base-in prism in the near segment of each lens (n = 46). Myopic progression measured by an automated refractor under cycloplegia and increase in axial length (secondary) measured by ultrasonography at 6-month intervals for 24 months. Only the data of the right eye were used. Of the 135 children (mean age, 10.29 years [SE, 0.15 years]; mean visual acuity, -3.08 D [SE, 0.10 D]), 131 (97%) completed the trial after 24 months. Myopic progression averaged -1.55 D (SE, 0.12 D) for those who wore single-vision lenses, -0.96 D (SE, 0.09 D) for those who wore bifocals, and -0.70 D (SE, 0.10 D) for those who wore prismatic bifocals. Axial length increased an average of 0.62 mm (SE, 0.04 mm), 0.41 mm (SE, 0.04 mm), and 0.41 mm (SE, 0.05 mm), respectively. The treatment effect of bifocals (0.59 D) and prismatic bifocals (0.85 D) was significant (P &lt; .001) and both bifocal groups had less axial elongation (0.21 mm) than the single-vision lens group (P &lt; .001). Bifocal lenses can moderately slow myopic progression in children with high rates of progression after 24 months. APPLICATIONS TO CLINICAL PRACTICE: Bifocal spectacles may be considered for slowing myopic progression in children with an annual progression rate of at least 0.50 D.

pubmed23n0743_7352

Induction of apoptosis of bladder cancer cells by zinc-citrate compound.

Zinc is one of the trace minerals in the body and is known to have an anticancer effect by inducing apoptosis in prostate cancer. We aimed to investigate the antiproliferative effects of a zinc-citrate compound in bladder cancer. A bladder cancer cell line (MBT-2) was treated with a zinc-citrate compound at different time intervals and concentrations. Mitochondrial (m)-aconitase activity was determined by use of the aconitase assay. DNA laddering analysis was performed to investigate apoptosis of MBT-2 cells. The molecular mechanism of apoptosis was investigated by Western blot analysis of p53, p21(waf1), Bcl-2, Bcl-xL, and Bax and also by caspase-3 activity analysis. Treatment with the zinc-citrate compound resulted in a time- and dose-dependent decrease in cell number of MBT-2 cells. M-aconitase activity was significantly decreased. DNA laddering analysis indicated apoptosis of MBT-2 cells. The zinc-citrate compound increased the expression of p21(waf1) and p53 and reduced the expression of Bcl-2 and Bcl-xL proteins but induced expression of Bax protein. The zinc-citrate compound induced apoptosis of MBT-2 cells by activation of the caspase-3 pathway. We have shown that a zinc-citrate compound induces apoptotic cell death in a bladder cancer cell line, MBT-2, by caspase-3 activation through up-regulation of apoptotic proteins and down-regulation of antiapoptotic proteins.

Induction of apoptosis of bladder cancer cells by zinc-citrate compound. Zinc is one of the trace minerals in the body and is known to have an anticancer effect by inducing apoptosis in prostate cancer. We aimed to investigate the antiproliferative effects of a zinc-citrate compound in bladder cancer. A bladder cancer cell line (MBT-2) was treated with a zinc-citrate compound at different time intervals and concentrations. Mitochondrial (m)-aconitase activity was determined by use of the aconitase assay. DNA laddering analysis was performed to investigate apoptosis of MBT-2 cells. The molecular mechanism of apoptosis was investigated by Western blot analysis of p53, p21(waf1), Bcl-2, Bcl-xL, and Bax and also by caspase-3 activity analysis. Treatment with the zinc-citrate compound resulted in a time- and dose-dependent decrease in cell number of MBT-2 cells. M-aconitase activity was significantly decreased. DNA laddering analysis indicated apoptosis of MBT-2 cells. The zinc-citrate compound increased the expression of p21(waf1) and p53 and reduced the expression of Bcl-2 and Bcl-xL proteins but induced expression of Bax protein. The zinc-citrate compound induced apoptosis of MBT-2 cells by activation of the caspase-3 pathway. We have shown that a zinc-citrate compound induces apoptotic cell death in a bladder cancer cell line, MBT-2, by caspase-3 activation through up-regulation of apoptotic proteins and down-regulation of antiapoptotic proteins.

pubmed23n0832_2429

Modified method of recording and reproducing natural head position with a multicamera system and a laser level.

As computer-assisted surgical design becomes increasingly popular in maxillofacial surgery, recording patients' natural head position (NHP) and reproducing it in the virtual environment are vital for preoperative design and postoperative evaluation. Our objective was to test the repeatability and accuracy of recording NHP using a multicamera system and a laser level. A laser level was used to project a horizontal reference line on a physical model, and a 3-dimensional image was obtained using a multicamera system. In surgical simulation software, the recorded NHP was reproduced in the virtual head position by registering the coordinate axes with the horizontal reference on both the frontal and lateral views. The repeatability and accuracy of the method were assessed using a gyroscopic procedure as the gold standard. The interclass correlation coefficients for pitch and roll were 0.982 (0.966, 0.991) and 0.995 (0.992, 0.998), respectively, indicating a high degree of repeatability. Regarding accuracy, the lack of agreement in orientation between the new method and the gold standard was within the ranges for pitch (-0.69°, 1.71°) and for roll (-0.92°, 1.20°); these have no clinical significance. This method of recording and reproducing NHP with a multicamera system and a laser level is repeatable, accurate, and clinically feasible.

Modified method of recording and reproducing natural head position with a multicamera system and a laser level. As computer-assisted surgical design becomes increasingly popular in maxillofacial surgery, recording patients' natural head position (NHP) and reproducing it in the virtual environment are vital for preoperative design and postoperative evaluation. Our objective was to test the repeatability and accuracy of recording NHP using a multicamera system and a laser level. A laser level was used to project a horizontal reference line on a physical model, and a 3-dimensional image was obtained using a multicamera system. In surgical simulation software, the recorded NHP was reproduced in the virtual head position by registering the coordinate axes with the horizontal reference on both the frontal and lateral views. The repeatability and accuracy of the method were assessed using a gyroscopic procedure as the gold standard. The interclass correlation coefficients for pitch and roll were 0.982 (0.966, 0.991) and 0.995 (0.992, 0.998), respectively, indicating a high degree of repeatability. Regarding accuracy, the lack of agreement in orientation between the new method and the gold standard was within the ranges for pitch (-0.69°, 1.71°) and for roll (-0.92°, 1.20°); these have no clinical significance. This method of recording and reproducing NHP with a multicamera system and a laser level is repeatable, accurate, and clinically feasible.

pubmed23n0104_7909

Cognitive spatial-motor processes. 2. Information transmitted by the direction of two-dimensional arm movements and by neuronal populations in primate motor cortex and area 5.

We measured the amount of information transmitted by the direction of two-dimensional (2-D) arm movements of human subjects, and by neuronal populations in the motor cortex and area 5 of monkeys. We also compared the information transmitted by a motor cortical population when the predictability of the direction of movement was varied, i.e. when the target of the movement was the same, or different, in successive trials. The information transmitted by a neuronal population was measured using the "population vector" code (Georgopoulos et al. 1983, 1986) as the stimulated directional output of the population. We found the following. (a) The information transmitted per movement increased in all cases with input information but more slowly than the maximum possible. No asymptote had been reached at 6.64 bits of input information; the best human performance at that level was 4.5 bits. (b) The average information transmitted by a motor cortical population (N = 253 cells) was approximately 0.5 bits higher than that of best human performance at all levels of input information. These findings indicate that whereas information loss at the level of motor cortex, and during stages preceding it, increases with increasing input information, this loss remains constant at all levels of input information during processes intervening between motor cortex and movement. (c) The information transmitted by the neuronal population at a constant level of input information increased with the number of cells in the population and had not plateaued at the population size of 253 cells used in this study. (d) Consistently higher amounts of information were transmitted by motor cortical than by area 5 populations at all levels of input information (up to 7 bits) examined. The difference was approximately 0.16 bits. (e) Finally, the information transmitted by a motor cortical population was slightly higher (by 0.07 bits) when the movement was more than less predictable. Although this difference was small, it was consistent at all levels of input information (3-7 bits) and was statistically highly significant (p less than 0.005, paired t test). These results establish the information theoretical approach as a useful method for comparing (a) behavioral and neural data, (b) neural data between different brain areas, and (c) neural data obtained under different behavioral manipulations.

Cognitive spatial-motor processes. 2. Information transmitted by the direction of two-dimensional arm movements and by neuronal populations in primate motor cortex and area 5. We measured the amount of information transmitted by the direction of two-dimensional (2-D) arm movements of human subjects, and by neuronal populations in the motor cortex and area 5 of monkeys. We also compared the information transmitted by a motor cortical population when the predictability of the direction of movement was varied, i.e. when the target of the movement was the same, or different, in successive trials. The information transmitted by a neuronal population was measured using the "population vector" code (Georgopoulos et al. 1983, 1986) as the stimulated directional output of the population. We found the following. (a) The information transmitted per movement increased in all cases with input information but more slowly than the maximum possible. No asymptote had been reached at 6.64 bits of input information; the best human performance at that level was 4.5 bits. (b) The average information transmitted by a motor cortical population (N = 253 cells) was approximately 0.5 bits higher than that of best human performance at all levels of input information. These findings indicate that whereas information loss at the level of motor cortex, and during stages preceding it, increases with increasing input information, this loss remains constant at all levels of input information during processes intervening between motor cortex and movement. (c) The information transmitted by the neuronal population at a constant level of input information increased with the number of cells in the population and had not plateaued at the population size of 253 cells used in this study. (d) Consistently higher amounts of information were transmitted by motor cortical than by area 5 populations at all levels of input information (up to 7 bits) examined. The difference was approximately 0.16 bits. (e) Finally, the information transmitted by a motor cortical population was slightly higher (by 0.07 bits) when the movement was more than less predictable. Although this difference was small, it was consistent at all levels of input information (3-7 bits) and was statistically highly significant (p less than 0.005, paired t test). These results establish the information theoretical approach as a useful method for comparing (a) behavioral and neural data, (b) neural data between different brain areas, and (c) neural data obtained under different behavioral manipulations.

pubmed23n0819_21587

Macroscopic anatomical, histological and magnetic resonance imaging correlation of the lateral capsule of the knee.

The objective of the present study was to correlate macroscopic and microscopic anatomy of the lateral capsule of the knee joint with high-quality magnetic resonance imaging (MRI), with a hypothesis that a distinct lateral capsular ligament would be inconsistently observed via surgical dissection and that high-quality MRI imaging would correlate to findings from dissection. Ten fresh-frozen human cadaveric knee specimens were utilized for this study. MRI of each knee was obtained pre- and post-dissection. The lateral knee was dissected and analysed for the presence or absence of a discrete capsular thickening or an independent ligamentous structure. A musculoskeletal radiologist analysed the pre- and post-dissection MRI. Subsequently, two specimens with positive lateral capsular thickening were prepared for histology. On macroscopic dissection, none of the ten specimens were found to have a discrete lateral capsular ligament. A palpable macroscopic thickening of the lateral capsule was identified in 4/10 specimens. MRI analysis revealed a 2-4 mm thickening of the central third of the lateral capsule in 3/10 specimens. On histological analysis, the lateral capsular thickening demonstrated properties similar to both capsule and ligament. In fresh-frozen cadaveric specimens, macroscopic and MRI evaluation of the lateral capsule of the knee revealed variations in morphology without consistent capsuloligamentous anatomy and specifically no discrete lateral capsular ligament. Further investigation in the form of clinical and mechanical relevance of the lateral capsular structures is of paramount importance before limited anatomical data can be utilized to drive clinical decision-making and patient care.

Macroscopic anatomical, histological and magnetic resonance imaging correlation of the lateral capsule of the knee. The objective of the present study was to correlate macroscopic and microscopic anatomy of the lateral capsule of the knee joint with high-quality magnetic resonance imaging (MRI), with a hypothesis that a distinct lateral capsular ligament would be inconsistently observed via surgical dissection and that high-quality MRI imaging would correlate to findings from dissection. Ten fresh-frozen human cadaveric knee specimens were utilized for this study. MRI of each knee was obtained pre- and post-dissection. The lateral knee was dissected and analysed for the presence or absence of a discrete capsular thickening or an independent ligamentous structure. A musculoskeletal radiologist analysed the pre- and post-dissection MRI. Subsequently, two specimens with positive lateral capsular thickening were prepared for histology. On macroscopic dissection, none of the ten specimens were found to have a discrete lateral capsular ligament. A palpable macroscopic thickening of the lateral capsule was identified in 4/10 specimens. MRI analysis revealed a 2-4 mm thickening of the central third of the lateral capsule in 3/10 specimens. On histological analysis, the lateral capsular thickening demonstrated properties similar to both capsule and ligament. In fresh-frozen cadaveric specimens, macroscopic and MRI evaluation of the lateral capsule of the knee revealed variations in morphology without consistent capsuloligamentous anatomy and specifically no discrete lateral capsular ligament. Further investigation in the form of clinical and mechanical relevance of the lateral capsular structures is of paramount importance before limited anatomical data can be utilized to drive clinical decision-making and patient care.

pubmed23n0376_14948

Genetic polymorphism of CYP2D6 and CYP2C19 in east- and southern African populations including psychiatric patients.

The study was carried out to investigate the distribution of cytochrome P450 2D6 (CYP2D6) and CYP2C19 genotype frequencies in three African populations and to compare these frequencies between healthy individuals and psychiatric patients. Three hundred and eighty-four subjects from South Africa (Venda), Tanzania, and Zimbabwe who consented to the study were genotyped for CYP2D6 (CYP2D6*1, *2, *3, *4, *5, and *17) and CYP2C19 (CYP2C19*1, *2, and *3) by PCR-RFLP (polymerase chain reaction restriction fragment length polymorphism) techniques. The genotypes for CYP2D6 predicted a poor metabolizer frequency of 2.3% (2/88) in Tanzanian psychiatric patients, 1.9% (2/106) in Tanzanian healthy controls and 2.6% (2/76) in the South African Venda. The low-activity CYP2D6*17 allele frequency was higher in psychiatric patients (30%, 53/176) than in healthy individuals (20%, 43/212) in Tanzanians. The frequencies for CYP2C19*2 genotypes were predictive of a low prevalence of poor metabolizers (PMs). The CYP2C19*3 allele was absent in the three populations studied. There was no difference in CYP2D6 or CYP2C19 PM genotype frequencies between psychiatric patients and healthy subjects. The genotype results predict a low prevalence of people with deficient CYP2D6 and CYP2C19 activity among linguistically (Bantu) related populations of East and Southern Africa. The high frequency of the low-activity CYP2D6*17 allele predicts that the Bantu people have a reduced capacity to metabolise drugs that are CYP2D6 substrates.

Genetic polymorphism of CYP2D6 and CYP2C19 in east- and southern African populations including psychiatric patients. The study was carried out to investigate the distribution of cytochrome P450 2D6 (CYP2D6) and CYP2C19 genotype frequencies in three African populations and to compare these frequencies between healthy individuals and psychiatric patients. Three hundred and eighty-four subjects from South Africa (Venda), Tanzania, and Zimbabwe who consented to the study were genotyped for CYP2D6 (CYP2D6*1, *2, *3, *4, *5, and *17) and CYP2C19 (CYP2C19*1, *2, and *3) by PCR-RFLP (polymerase chain reaction restriction fragment length polymorphism) techniques. The genotypes for CYP2D6 predicted a poor metabolizer frequency of 2.3% (2/88) in Tanzanian psychiatric patients, 1.9% (2/106) in Tanzanian healthy controls and 2.6% (2/76) in the South African Venda. The low-activity CYP2D6*17 allele frequency was higher in psychiatric patients (30%, 53/176) than in healthy individuals (20%, 43/212) in Tanzanians. The frequencies for CYP2C19*2 genotypes were predictive of a low prevalence of poor metabolizers (PMs). The CYP2C19*3 allele was absent in the three populations studied. There was no difference in CYP2D6 or CYP2C19 PM genotype frequencies between psychiatric patients and healthy subjects. The genotype results predict a low prevalence of people with deficient CYP2D6 and CYP2C19 activity among linguistically (Bantu) related populations of East and Southern Africa. The high frequency of the low-activity CYP2D6*17 allele predicts that the Bantu people have a reduced capacity to metabolise drugs that are CYP2D6 substrates.

pubmed23n0819_9848

Retrieval capabilities of hierarchical networks: from Dyson to Hopfield.

We consider statistical-mechanics models for spin systems built on hierarchical structures, which provide a simple example of non-mean-field framework. We show that the coupling decay with spin distance can give rise to peculiar features and phase diagrams much richer than their mean-field counterpart. In particular, we consider the Dyson model, mimicking ferromagnetism in lattices, and we prove the existence of a number of metastabilities, beyond the ordered state, which become stable in the thermodynamic limit. Such a feature is retained when the hierarchical structure is coupled with the Hebb rule for learning, hence mimicking the modular architecture of neurons, and gives rise to an associative network able to perform single pattern retrieval as well as multiple-pattern retrieval, depending crucially on the external stimuli and on the rate of interaction decay with distance; however, those emergent multitasking features reduce the network capacity with respect to the mean-field counterpart. The analysis is accomplished through statistical mechanics, Markov chain theory, signal-to-noise ratio technique, and numerical simulations in full consistency. Our results shed light on the biological complexity shown by real networks, and suggest future directions for understanding more realistic models.

Retrieval capabilities of hierarchical networks: from Dyson to Hopfield. We consider statistical-mechanics models for spin systems built on hierarchical structures, which provide a simple example of non-mean-field framework. We show that the coupling decay with spin distance can give rise to peculiar features and phase diagrams much richer than their mean-field counterpart. In particular, we consider the Dyson model, mimicking ferromagnetism in lattices, and we prove the existence of a number of metastabilities, beyond the ordered state, which become stable in the thermodynamic limit. Such a feature is retained when the hierarchical structure is coupled with the Hebb rule for learning, hence mimicking the modular architecture of neurons, and gives rise to an associative network able to perform single pattern retrieval as well as multiple-pattern retrieval, depending crucially on the external stimuli and on the rate of interaction decay with distance; however, those emergent multitasking features reduce the network capacity with respect to the mean-field counterpart. The analysis is accomplished through statistical mechanics, Markov chain theory, signal-to-noise ratio technique, and numerical simulations in full consistency. Our results shed light on the biological complexity shown by real networks, and suggest future directions for understanding more realistic models.

pubmed23n1048_14591

Structure of a human 48<i>S</i> translational initiation complex.

A key step in translational initiation is the recruitment of the 43<iS</i preinitiation complex by the cap-binding complex [eukaryotic initiation factor 4F (eIF4F)] at the 5' end of messenger RNA (mRNA) to form the 48<iS</i initiation complex (i.e., the 48<iS</i). The 48<iS</i then scans along the mRNA to locate a start codon. To understand the mechanisms involved, we used cryo-electron microscopy to determine the structure of a reconstituted human 48<iS</i The structure reveals insights into early events of translation initiation complex assembly, as well as how eIF4F interacts with subunits of eIF3 near the mRNA exit channel in the 43<iS</i The location of eIF4F is consistent with a slotting model of mRNA recruitment and suggests that downstream mRNA is unwound at least in part by being "pulled" through the 40<iS</i subunit during scanning.

Structure of a human 48<i>S</i> translational initiation complex. A key step in translational initiation is the recruitment of the 43<iS</i preinitiation complex by the cap-binding complex [eukaryotic initiation factor 4F (eIF4F)] at the 5' end of messenger RNA (mRNA) to form the 48<iS</i initiation complex (i.e., the 48<iS</i). The 48<iS</i then scans along the mRNA to locate a start codon. To understand the mechanisms involved, we used cryo-electron microscopy to determine the structure of a reconstituted human 48<iS</i The structure reveals insights into early events of translation initiation complex assembly, as well as how eIF4F interacts with subunits of eIF3 near the mRNA exit channel in the 43<iS</i The location of eIF4F is consistent with a slotting model of mRNA recruitment and suggests that downstream mRNA is unwound at least in part by being "pulled" through the 40<iS</i subunit during scanning.

pubmed23n1075_8433

Factors associated with elevated low-density lipoprotein cholesterol levels among hill tribe people aged 30 years and over in Thailand: a cross-sectional study.

Low-density lipoprotein cholesterol (LDL-C) is one of the most important types of cholesterol and has an impact on health. Certain lifestyle and dietary habits in different populations may leads to increased levels of LDL-C, particularly among those with poor education and economic statuses, such as hill tribe people in Thailand. This study aimed to estimate the prevalence of and determine the factors associated with high LDL-C levels among hill tribe people in northern Thailand. A cross-sectional study was performed to gather information from six main hill tribe populations: Akha, Lahu, Hmong, Yao, Karen, and Lisu. Individuals who were aged over 30 years and living in 30 selected hill tribe villages were invited to participate in the study. A validated questionnaire and 5-mL blood specimens were used to obtain data. Correlation analyses, chi-square tests, t-tests, and logistic regression were used to detect correlations and associations. A total of 2552 participants were recruited into the study; 65.9% were females, and 64.1% were aged younger than 60 years old. Approximately 69.6% of participants had abnormal LDL-C levels; 33.6% had above-optimal levels, 24.3% had borderline high levels, 8.0% had high levels, and 3.7% had very high levels. A total of 17.4% of participants had low high-density lipoprotein cholesterol (HDL-C) and high LDL-C levels, while 14.9% had high triglyceride and LDL-C levels. After controlling for sex, age, religion, education, annual family income, and marital status in the multivariate model, three variables were found to be associated with high LDL-C levels: occupation, the amount of lard used in daily cooking, and glycated hemoglobin (HbA1c). Those who were working as agriculturalists had a 1.34-fold greater chance of having abnormal LDL-C than traders and others (95% CI = 1.09-1.34). Those who used moderate and high quantities of lard in their daily cooking had a 1.45-fold (95% CI = 1.15-1.82) and 1.31-fold (95% CI = 1.04-1.68) greater likelihood of having abnormal LDL-C levels than those who used low quantities, respectively. Those who had abnormal HbA1c levels were less likely to develop abnormal LDL-C levels than those who had normal HbA1c levels (AOR = 0.69, 95% CI = 0.51-92). Effective public health programs that do not conflict with the cultures of hill tribes are urgently needed, particularly programs encouraging the use of small quantities of lard for daily cooking practices.

Factors associated with elevated low-density lipoprotein cholesterol levels among hill tribe people aged 30 years and over in Thailand: a cross-sectional study. Low-density lipoprotein cholesterol (LDL-C) is one of the most important types of cholesterol and has an impact on health. Certain lifestyle and dietary habits in different populations may leads to increased levels of LDL-C, particularly among those with poor education and economic statuses, such as hill tribe people in Thailand. This study aimed to estimate the prevalence of and determine the factors associated with high LDL-C levels among hill tribe people in northern Thailand. A cross-sectional study was performed to gather information from six main hill tribe populations: Akha, Lahu, Hmong, Yao, Karen, and Lisu. Individuals who were aged over 30 years and living in 30 selected hill tribe villages were invited to participate in the study. A validated questionnaire and 5-mL blood specimens were used to obtain data. Correlation analyses, chi-square tests, t-tests, and logistic regression were used to detect correlations and associations. A total of 2552 participants were recruited into the study; 65.9% were females, and 64.1% were aged younger than 60 years old. Approximately 69.6% of participants had abnormal LDL-C levels; 33.6% had above-optimal levels, 24.3% had borderline high levels, 8.0% had high levels, and 3.7% had very high levels. A total of 17.4% of participants had low high-density lipoprotein cholesterol (HDL-C) and high LDL-C levels, while 14.9% had high triglyceride and LDL-C levels. After controlling for sex, age, religion, education, annual family income, and marital status in the multivariate model, three variables were found to be associated with high LDL-C levels: occupation, the amount of lard used in daily cooking, and glycated hemoglobin (HbA1c). Those who were working as agriculturalists had a 1.34-fold greater chance of having abnormal LDL-C than traders and others (95% CI = 1.09-1.34). Those who used moderate and high quantities of lard in their daily cooking had a 1.45-fold (95% CI = 1.15-1.82) and 1.31-fold (95% CI = 1.04-1.68) greater likelihood of having abnormal LDL-C levels than those who used low quantities, respectively. Those who had abnormal HbA1c levels were less likely to develop abnormal LDL-C levels than those who had normal HbA1c levels (AOR = 0.69, 95% CI = 0.51-92). Effective public health programs that do not conflict with the cultures of hill tribes are urgently needed, particularly programs encouraging the use of small quantities of lard for daily cooking practices.

pubmed23n0623_17547

Olanzapine treatment in anorexia nervosa: case report.

A 15 year old patient suffering from psychiatric disturbances looked for psychiatric help but refused hospital admission. Following an ambulatory treatment, the patient was diagnosed with Anorexia nervosa. The patient, a girl, was 175 centimeters tall, weighting only 39 kilos. Within the clinical picture, there were few dominant disorders present; anxiety, depression, low self-esteem, fear of feminization, with recurrent psychotic episodes. By the implementation of an intensive psychotherapeutic treatment, without the use of psychopharmacs, the weight was kept stable. In accordance with the girl's mother, a psychopharmacotherapy was commenced, a combination of olanzapine and paroxetine (the choice of psychopharmacs was lead by the side effects known). At the end of a 24-month period of a psychological treatment which was combined with psychopharmacotherapy, the patient exhibited no symptomatology and a stable clinical remission of the illness was achieved.

Olanzapine treatment in anorexia nervosa: case report. A 15 year old patient suffering from psychiatric disturbances looked for psychiatric help but refused hospital admission. Following an ambulatory treatment, the patient was diagnosed with Anorexia nervosa. The patient, a girl, was 175 centimeters tall, weighting only 39 kilos. Within the clinical picture, there were few dominant disorders present; anxiety, depression, low self-esteem, fear of feminization, with recurrent psychotic episodes. By the implementation of an intensive psychotherapeutic treatment, without the use of psychopharmacs, the weight was kept stable. In accordance with the girl's mother, a psychopharmacotherapy was commenced, a combination of olanzapine and paroxetine (the choice of psychopharmacs was lead by the side effects known). At the end of a 24-month period of a psychological treatment which was combined with psychopharmacotherapy, the patient exhibited no symptomatology and a stable clinical remission of the illness was achieved.

pubmed23n0257_16627

Influence of valine flooding on channeling of valine into tissue pools and on protein synthesis.

Rates of valine incorporation into protein were measured under control and valine-"flooding" conditions and included correction for the degree of recycling of unlabeled valine derived from the steady-state breakdown of tissue protein into the precursor pool (tRNA bound). The correction factor lambda, which is the ratio of the steady-state specific activity of valine in the tissue tRNA-bound pool to that in the arterial plasma, was determined for each of the tissues. In controls, values of lambda ranged from 0.31 in adrenals to 0.54 in heart; in flooded animals, values were higher, but only in liver was the value of lambda close to 1.0. In control and flooded rats, rates of protein synthesis were highest in liver and adrenals and lowest in skeletal muscle, with intermediate values in brain and heart. Flooding resulted in increased rates of protein synthesis in liver and decreased rates in adrenals. Rates of protein synthesis in brain, heart, and skeletal muscle were not statistically significantly affected by flooding.

Influence of valine flooding on channeling of valine into tissue pools and on protein synthesis. Rates of valine incorporation into protein were measured under control and valine-"flooding" conditions and included correction for the degree of recycling of unlabeled valine derived from the steady-state breakdown of tissue protein into the precursor pool (tRNA bound). The correction factor lambda, which is the ratio of the steady-state specific activity of valine in the tissue tRNA-bound pool to that in the arterial plasma, was determined for each of the tissues. In controls, values of lambda ranged from 0.31 in adrenals to 0.54 in heart; in flooded animals, values were higher, but only in liver was the value of lambda close to 1.0. In control and flooded rats, rates of protein synthesis were highest in liver and adrenals and lowest in skeletal muscle, with intermediate values in brain and heart. Flooding resulted in increased rates of protein synthesis in liver and decreased rates in adrenals. Rates of protein synthesis in brain, heart, and skeletal muscle were not statistically significantly affected by flooding.

pubmed23n0917_12088

[Clinical analysis of orbital and periorbital abscess in 17 patients].

<bObjective:</b To discuss the etiology, clinical features and treatment principles of the orbital and periorbital abscess. <bMethods:</b A retrospective case series of 17 cases with orbital and periorbital abscess between July 2010 and November 2015 were conducted. All patients(,) clinical data including medical history, etiology, abscess location, paranasal sinus involvement, eye involvement, microbiological test results, imaging features, treatment and prognosis were summarized and analyzed. <bResults:</b In all 17 patients, there were 10 males and 7 females with age from 3.0 to 71.0 years (the average age was 33.9 years).Eight patients(,) bacterial cultures of the pus and secretion were positive in all 17 patients. Orbital and periorbital abscess patients could manifest decreased vision, redness and swelling of eyelid, conjunctival congestion and edema, ocular motility disorders, displacement of eyeball, increased orbital pressure, abscess rupture etc. CT showed us the soft tissue mass, accompanied with sinusitis or paranasal sinus mass. MR performed with the long T(1) and T(2) signals. The signals of the abscess cavity were not uniform. For the etiology,11 cases were secondary to sinusitis, including 1 case of diabetes; 2 cases with orbital fractures.One case was secondary to orbital fracture repairment surgery. One case was secondary to the remnant of sequestrum and foreign bodys in the wound after repairment surgery. One case was injured by the hard object. One case was secondary to paranasal sinuses large B-cell lymphoma. One case had diabetic history and the blood sugar was controlled unstablly. For the treatment, 7 cases were treated by the drainage surgery which was performed via the sinus with endoscopic and abscess resection performed via the skin.Two cases were treatment by the abscess resection only.One case was treated by the drainage surgery performed via the sinus with endoscopic only. Six cases were treated by the drainage surgery performed via the skin. One case was only administered intravenous antibiotic. Sixteen cases acquired well prognosis without serious complications except 1 case which occurred central retinal artery and vein occlusion. <bConclusions:</b The orbital and periorbital abscess is mainly a complication of paranasal sinus infection, or secondary to trauma, surgery, tumor, etc; Orbital and periorbital abscess always manifest inflammatory neoplastic clinical features, the key of the diagnosis is to make sure the etiology; Incision and drainage of the abscess is the main treatment method when necessary. We can do the surgery with other departments to avoid the occurrence of serious complications.<i(Chin J Ophthalmol, 2017, 53:</i 588-593<i)</i.

[Clinical analysis of orbital and periorbital abscess in 17 patients]. <bObjective:</b To discuss the etiology, clinical features and treatment principles of the orbital and periorbital abscess. <bMethods:</b A retrospective case series of 17 cases with orbital and periorbital abscess between July 2010 and November 2015 were conducted. All patients(,) clinical data including medical history, etiology, abscess location, paranasal sinus involvement, eye involvement, microbiological test results, imaging features, treatment and prognosis were summarized and analyzed. <bResults:</b In all 17 patients, there were 10 males and 7 females with age from 3.0 to 71.0 years (the average age was 33.9 years).Eight patients(,) bacterial cultures of the pus and secretion were positive in all 17 patients. Orbital and periorbital abscess patients could manifest decreased vision, redness and swelling of eyelid, conjunctival congestion and edema, ocular motility disorders, displacement of eyeball, increased orbital pressure, abscess rupture etc. CT showed us the soft tissue mass, accompanied with sinusitis or paranasal sinus mass. MR performed with the long T(1) and T(2) signals. The signals of the abscess cavity were not uniform. For the etiology,11 cases were secondary to sinusitis, including 1 case of diabetes; 2 cases with orbital fractures.One case was secondary to orbital fracture repairment surgery. One case was secondary to the remnant of sequestrum and foreign bodys in the wound after repairment surgery. One case was injured by the hard object. One case was secondary to paranasal sinuses large B-cell lymphoma. One case had diabetic history and the blood sugar was controlled unstablly. For the treatment, 7 cases were treated by the drainage surgery which was performed via the sinus with endoscopic and abscess resection performed via the skin.Two cases were treatment by the abscess resection only.One case was treated by the drainage surgery performed via the sinus with endoscopic only. Six cases were treated by the drainage surgery performed via the skin. One case was only administered intravenous antibiotic. Sixteen cases acquired well prognosis without serious complications except 1 case which occurred central retinal artery and vein occlusion. <bConclusions:</b The orbital and periorbital abscess is mainly a complication of paranasal sinus infection, or secondary to trauma, surgery, tumor, etc; Orbital and periorbital abscess always manifest inflammatory neoplastic clinical features, the key of the diagnosis is to make sure the etiology; Incision and drainage of the abscess is the main treatment method when necessary. We can do the surgery with other departments to avoid the occurrence of serious complications.<i(Chin J Ophthalmol, 2017, 53:</i 588-593<i)</i.

pubmed23n0585_809

High grade glioma: imaging combined with pathological grade defines management and predicts prognosis.

There is ambiguity in pathological grading of high grade gliomas within the WHO 2000 classification, especially those with predominant oligodendroglial differentiation. All adult high grade gliomas treated radically, 1996-2005, were assessed. Cases in which pathology was grade III but radiology suggested glioblastoma (GBM) were classified as 'grade III/IV'; their pathology was reviewed. Data from 245 patients (52 grade III, 18 grade III/IV, 175 GBM) were analysed using a Cox Proportional Hazards model. On pathology review, features suggestive of more aggressive behaviour were found in all 18 grade III/IV tumours. Oligodendroglial components with both necrosis and microvascular proliferation were present in 7. MIB-1 counts for the last 8 were all above 14%, mean 27%. Median survivals were: grade III 34 months, grade III/IV 10 months, GBM 11 months. Survival was not significantly different between grade III/IV and GBM. Patients with grade III/IV tumours had significantly worse outcome than grade III, with a hazard of death 3.7 times higher. The results highlight the current inconsistency in pathological grading of high grade tumours, especially those with oligodendroglial elements. Patients with histological grade III tumours but radiological appearances suggestive of GBM should be managed as glioblastoma.

High grade glioma: imaging combined with pathological grade defines management and predicts prognosis. There is ambiguity in pathological grading of high grade gliomas within the WHO 2000 classification, especially those with predominant oligodendroglial differentiation. All adult high grade gliomas treated radically, 1996-2005, were assessed. Cases in which pathology was grade III but radiology suggested glioblastoma (GBM) were classified as 'grade III/IV'; their pathology was reviewed. Data from 245 patients (52 grade III, 18 grade III/IV, 175 GBM) were analysed using a Cox Proportional Hazards model. On pathology review, features suggestive of more aggressive behaviour were found in all 18 grade III/IV tumours. Oligodendroglial components with both necrosis and microvascular proliferation were present in 7. MIB-1 counts for the last 8 were all above 14%, mean 27%. Median survivals were: grade III 34 months, grade III/IV 10 months, GBM 11 months. Survival was not significantly different between grade III/IV and GBM. Patients with grade III/IV tumours had significantly worse outcome than grade III, with a hazard of death 3.7 times higher. The results highlight the current inconsistency in pathological grading of high grade tumours, especially those with oligodendroglial elements. Patients with histological grade III tumours but radiological appearances suggestive of GBM should be managed as glioblastoma.

pubmed23n1023_1893

Effects of staphylococcal infection and aseptic inflammation on bone mass and biomechanical properties in a rabbit model.

Orthopaedic implants are important devices aimed at relieving pain and improving mobility. Staphylococcal infection and aseptic loosening are two common events associated with inflammatory osteolysis that lead to implant failures. Bone mass and biomechanical properties are important indicators that could influence patient outcomes after revision surgery. However, the dynamics of bacterial infections and their influence on bone mass and biomechanical properties remain unclear. Hence, in this study, we developed rabbit aseptic inflammation and staphylococcal infection models to determine the effects of coagulase-positive and coagulase-negative bacterial infection, as well as aseptic inflammation, on the mass and biomechanical properties of the bone. Sixty New Zealand white rabbits were randomly assigned to 6 groups, and each group had 10 rabbits. The medullary cavities in rabbits of each group were injected with phosphate-buffered saline (100 ​μL), titanium (Ti)-wear particles (300 μg/100 ​μL), a low concentration of <iStaphylococcus epidermidis</i (10<sup5</sup/100 ​μL), a high concentration of <iS. epidermidis</i (10<sup8</sup/100 ​μL), a low concentration of <iStaphylococcus aureus</i (10<sup5</sup/100 ​μL), and a high concentration of <iS. aureus</i (10<sup8</sup/100 ​μL), respectively. At four and eight weeks after surgery, the rabbits were sacrificed, and the tibias on the surgical side were analysed via histopathology, microcomputed tomography, and nanoindentation testing. Histopathological analysis demonstrated that inflammatory responses and bacterial loads caused by high concentrations of staphylococcal infections, particularly coagulase-positive staphylococci, are more detrimental than low concentrations of bacterial infection and Ti-wear particles. Meanwhile, microcomputed tomography and nanoindentation testing showed that high concentrations of <iS. aureus</i caused the highest loss in bone mass and most biomechanical function impairment in rabbits experiencing aseptic inflammation and staphylococcal infections. Inflammatory osteolysis caused by a high concentration of coagulase-positive staphylococci is significantly associated with low bone mass and impaired biomechanical properties. It is necessary to obtain an overall assessment of the bone mass and biomechanical properties before revision surgery, especially when <iS. aureus</i infection is involved. In addition, a better understanding of these two parameters might help develop a reasonable treatment regimen and reduce the risk of adverse events after revision surgery.

Effects of staphylococcal infection and aseptic inflammation on bone mass and biomechanical properties in a rabbit model. Orthopaedic implants are important devices aimed at relieving pain and improving mobility. Staphylococcal infection and aseptic loosening are two common events associated with inflammatory osteolysis that lead to implant failures. Bone mass and biomechanical properties are important indicators that could influence patient outcomes after revision surgery. However, the dynamics of bacterial infections and their influence on bone mass and biomechanical properties remain unclear. Hence, in this study, we developed rabbit aseptic inflammation and staphylococcal infection models to determine the effects of coagulase-positive and coagulase-negative bacterial infection, as well as aseptic inflammation, on the mass and biomechanical properties of the bone. Sixty New Zealand white rabbits were randomly assigned to 6 groups, and each group had 10 rabbits. The medullary cavities in rabbits of each group were injected with phosphate-buffered saline (100 ​μL), titanium (Ti)-wear particles (300 μg/100 ​μL), a low concentration of <iStaphylococcus epidermidis</i (10<sup5</sup/100 ​μL), a high concentration of <iS. epidermidis</i (10<sup8</sup/100 ​μL), a low concentration of <iStaphylococcus aureus</i (10<sup5</sup/100 ​μL), and a high concentration of <iS. aureus</i (10<sup8</sup/100 ​μL), respectively. At four and eight weeks after surgery, the rabbits were sacrificed, and the tibias on the surgical side were analysed via histopathology, microcomputed tomography, and nanoindentation testing. Histopathological analysis demonstrated that inflammatory responses and bacterial loads caused by high concentrations of staphylococcal infections, particularly coagulase-positive staphylococci, are more detrimental than low concentrations of bacterial infection and Ti-wear particles. Meanwhile, microcomputed tomography and nanoindentation testing showed that high concentrations of <iS. aureus</i caused the highest loss in bone mass and most biomechanical function impairment in rabbits experiencing aseptic inflammation and staphylococcal infections. Inflammatory osteolysis caused by a high concentration of coagulase-positive staphylococci is significantly associated with low bone mass and impaired biomechanical properties. It is necessary to obtain an overall assessment of the bone mass and biomechanical properties before revision surgery, especially when <iS. aureus</i infection is involved. In addition, a better understanding of these two parameters might help develop a reasonable treatment regimen and reduce the risk of adverse events after revision surgery.

pubmed23n0723_12534

Dialect experience and perceptual integrality in phonological registers: fundamental frequency, voice quality and the first formant in Cham.

The perceptual integrality of f0, F1 and voice quality is investigated by looking at register, a phonological contrast that relies on these three properties in three dialects of Cham, an Austronesian language of Mainland Southeast Asia. The results of a Garner classification experiment confirm that the three acoustic properties integrate perceptually and that their patterns of integrality are similar in the three dialects. Moreover, they show that dialect-specific sensitivity to acoustic properties can cause salient dimensions to override weaker ones. Finally, the patterns of integrality found in Cham suggest that auditory integrality is not limited to acoustically similar properties.

Dialect experience and perceptual integrality in phonological registers: fundamental frequency, voice quality and the first formant in Cham. The perceptual integrality of f0, F1 and voice quality is investigated by looking at register, a phonological contrast that relies on these three properties in three dialects of Cham, an Austronesian language of Mainland Southeast Asia. The results of a Garner classification experiment confirm that the three acoustic properties integrate perceptually and that their patterns of integrality are similar in the three dialects. Moreover, they show that dialect-specific sensitivity to acoustic properties can cause salient dimensions to override weaker ones. Finally, the patterns of integrality found in Cham suggest that auditory integrality is not limited to acoustically similar properties.

pubmed23n0686_6117

ABO blood group alleles and the risk of pancreatic cancer in a Japanese population.

Several studies have investigated a possible association between the ABO blood group and the risk of pancreatic cancer (PC), but this association has not been fully evaluated in Asian populations. The present study aimed to assess the impact of genotype-derived ABO blood types, particularly ABO alleles, on the risk of PC in a Japanese population. We conducted a case-control study using 185 PC and 1465 control patients who visited Aichi Cancer Center in Nagoya, Japan. Using rs8176719 as a marker for the O allele, and rs8176746 and rs8176747 for the B allele, all participants' two ABO alleles were inferred. The impact of ABO blood type on PC risk was examined by multivariate analysis, with adjustment for potential confounders to estimate odds ratios (OR) and 95% confidence intervals (CI). An increased risk of PC was observed with the addition of any non-O allele (trend P = 0.012). Compared with subjects with the OO genotype, those with AO and BB genotypes had significantly increased OR of 1.67 (CI, 1.08-2.57) and 3.28 (CI, 1.38-7.80), respectively. Consistent with earlier reports showing a higher risk of PC for individuals with the non-O blood type, the previously reported protective allele (T) for rs505922 was found to be strongly correlated (r(2) = 0.96) with the O allele. In conclusion, this case-control study showed a statistically significant association between ABO blood group and PC risk in a Japanese population. Further studies are necessary to define the mechanisms by which the ABO gene or closely linked genetic variants influence PC risk.

ABO blood group alleles and the risk of pancreatic cancer in a Japanese population. Several studies have investigated a possible association between the ABO blood group and the risk of pancreatic cancer (PC), but this association has not been fully evaluated in Asian populations. The present study aimed to assess the impact of genotype-derived ABO blood types, particularly ABO alleles, on the risk of PC in a Japanese population. We conducted a case-control study using 185 PC and 1465 control patients who visited Aichi Cancer Center in Nagoya, Japan. Using rs8176719 as a marker for the O allele, and rs8176746 and rs8176747 for the B allele, all participants' two ABO alleles were inferred. The impact of ABO blood type on PC risk was examined by multivariate analysis, with adjustment for potential confounders to estimate odds ratios (OR) and 95% confidence intervals (CI). An increased risk of PC was observed with the addition of any non-O allele (trend P = 0.012). Compared with subjects with the OO genotype, those with AO and BB genotypes had significantly increased OR of 1.67 (CI, 1.08-2.57) and 3.28 (CI, 1.38-7.80), respectively. Consistent with earlier reports showing a higher risk of PC for individuals with the non-O blood type, the previously reported protective allele (T) for rs505922 was found to be strongly correlated (r(2) = 0.96) with the O allele. In conclusion, this case-control study showed a statistically significant association between ABO blood group and PC risk in a Japanese population. Further studies are necessary to define the mechanisms by which the ABO gene or closely linked genetic variants influence PC risk.

pubmed23n1023_25469

Participant incentives in rehabilitation research: a pilot study of researchers' perspectives.

Despite the use of research incentives in the recruitment and retention of participants for research studies, there remains debate over the acceptable use of participant incentives in research. There appears to be a paucity of guidelines that can assist researchers in demonstrating practically how incentives may be ethically used in research. This single site pilot study explored the experiences of key stakeholders involved in rehabilitation research to highlight what may constitute acceptable practices for incentives. A qualitative inquiry with use of semi-structured interviews with four key informants from a single site was undertaken. Data was audio-recorded and analysed thematically using deductive reasoning. The findings reflect a description of what incentives constitute; the issues around undue inducement and use of incentives in practice. Participants' offered their perceptions on perceived acceptable versus unacceptable practices in the use incentives in research. Participants shared their concern over research incentives being used to sway participation in research rather than reimburse participants for their expenses or offering a token of appreciation. There is a need for education of practitioners and researchers to develop skill to aid researchers to evaluate the ethical dilemmas related to the use of incentives in research.

Participant incentives in rehabilitation research: a pilot study of researchers' perspectives. Despite the use of research incentives in the recruitment and retention of participants for research studies, there remains debate over the acceptable use of participant incentives in research. There appears to be a paucity of guidelines that can assist researchers in demonstrating practically how incentives may be ethically used in research. This single site pilot study explored the experiences of key stakeholders involved in rehabilitation research to highlight what may constitute acceptable practices for incentives. A qualitative inquiry with use of semi-structured interviews with four key informants from a single site was undertaken. Data was audio-recorded and analysed thematically using deductive reasoning. The findings reflect a description of what incentives constitute; the issues around undue inducement and use of incentives in practice. Participants' offered their perceptions on perceived acceptable versus unacceptable practices in the use incentives in research. Participants shared their concern over research incentives being used to sway participation in research rather than reimburse participants for their expenses or offering a token of appreciation. There is a need for education of practitioners and researchers to develop skill to aid researchers to evaluate the ethical dilemmas related to the use of incentives in research.

pubmed23n0326_17721

Dioctahedral smectite neutralization activity of Clostridium difficile and Bacteroides fragilis toxins in vitro.

The neutralization activity of dioctahedral smectite for ten toxigenic Clostridium difficile and eight enterotoxigenic Bacteroides fragilis strains was studied using McCoy and HT 29/C1 cell lines, respectively. Minimalization of the cytopathic effect of C. difficile toxin B on McCoy cell lines by dioctahedral smectite dissolved in PBS was observed. After incubation with dioctahedral smectite the toxic effects of B. fragilis enterotoxins on HT/29C1 (human colon adenocarcinoma cell line) were eliminated. Best neutralization of B. fragilis toxin was achieved using dioctahedral smectite dissolved in BHI.

Dioctahedral smectite neutralization activity of Clostridium difficile and Bacteroides fragilis toxins in vitro. The neutralization activity of dioctahedral smectite for ten toxigenic Clostridium difficile and eight enterotoxigenic Bacteroides fragilis strains was studied using McCoy and HT 29/C1 cell lines, respectively. Minimalization of the cytopathic effect of C. difficile toxin B on McCoy cell lines by dioctahedral smectite dissolved in PBS was observed. After incubation with dioctahedral smectite the toxic effects of B. fragilis enterotoxins on HT/29C1 (human colon adenocarcinoma cell line) were eliminated. Best neutralization of B. fragilis toxin was achieved using dioctahedral smectite dissolved in BHI.

pubmed23n0666_16696

Zebrafish (Danio rerio) vary by strain and sex in their behavioral and transcriptional responses to selenium supplementation.

We used the Nadia, Gaighatta, Scientific Hatcheries, and TM1 zebrafish (Danio rerio) strains to test the hypothesis that variation among populations influences the behavioral and transcriptional responses to selenium supplementation. When fed a diet with control levels of selenium, zebrafish strains differed significantly in behavior, characterized as their mean horizontal and vertical swimming positions within the tank. The four strains also differed in brain expression of selenoprotein P1a (sepp1a), glutathione peroxidase 3 (gpx3), thioredoxin reductase 1 (txnrd1), and tRNA selenocysteine associated protein 1 (secp43). Iodothyronine deiodinase 2 (dio2) did not differ among strains but showed a sex-specific expression pattern. When supplemented with selenium, all strains spent a greater proportion of time near the front of the tank, but the response of vertical swimming depth varied by strain. Selenium supplementation also caused changes in selenoprotein expression in the brain that varied by strain for sepp1a, secp43, and dio2, and varied by strain and sex for txnrd1. Expression of gpx3 was unaffected by selenium. Our data indicate that selenium homeostasis in the brain may be a regulator of behavior in zebrafish, and the strain-specific effects of selenium supplementation suggest that genetic heterogeneity among populations can influence the results of selenium supplementation studies.

Zebrafish (Danio rerio) vary by strain and sex in their behavioral and transcriptional responses to selenium supplementation. We used the Nadia, Gaighatta, Scientific Hatcheries, and TM1 zebrafish (Danio rerio) strains to test the hypothesis that variation among populations influences the behavioral and transcriptional responses to selenium supplementation. When fed a diet with control levels of selenium, zebrafish strains differed significantly in behavior, characterized as their mean horizontal and vertical swimming positions within the tank. The four strains also differed in brain expression of selenoprotein P1a (sepp1a), glutathione peroxidase 3 (gpx3), thioredoxin reductase 1 (txnrd1), and tRNA selenocysteine associated protein 1 (secp43). Iodothyronine deiodinase 2 (dio2) did not differ among strains but showed a sex-specific expression pattern. When supplemented with selenium, all strains spent a greater proportion of time near the front of the tank, but the response of vertical swimming depth varied by strain. Selenium supplementation also caused changes in selenoprotein expression in the brain that varied by strain for sepp1a, secp43, and dio2, and varied by strain and sex for txnrd1. Expression of gpx3 was unaffected by selenium. Our data indicate that selenium homeostasis in the brain may be a regulator of behavior in zebrafish, and the strain-specific effects of selenium supplementation suggest that genetic heterogeneity among populations can influence the results of selenium supplementation studies.

pubmed23n0077_10520

Mechanistic investigation of doxycycline photosensitization by picosecond-pulsed and continuous wave laser irradiation of cells in culture.

In order to elucidate the photophysical mechanisms of cellular phototoxicity sensitized by doxycycline, MGH-U1 human bladder carcinoma cells in vitro were treated with 20.7 microM doxycycline and irradiated with either a pulsed (lambda = 355 nm, pulse duration = 24 ps) or a continuous wave (lambda = 351 nm) laser. Cumulative radiant exposure and irradiance were systematically varied in experiments with both lasers. Phototoxicity was assessed by epifluorescence microscopy of unfixed cells using rhodamine 123 labeling of mitochondria. With the continuous wave source, the cumulative radiant exposure required for induction of phototoxic injury was independent of irradiance. With the 24-ps-pulsed source, a significantly lower cumulative radiant exposure was required to induce the phototoxicity when the peak irradiance was 5.8 x 10(7) or 1.3 x 10(8) watts cm-2 compared with when peak irradiance was either lower (6.0 x 10(6) watts cm-2) or higher (7.6 x 10(8) watts cm-2). The measured fluorescence lifetimes of doxycycline in buffered saline solution were longer than the laser pulse duration of 24 ps. The increased efficiency of photosensitization at the optimal peak irradiance in the ps domain appears to result from sequential multiphoton absorption involving higher excited states of the singlet manifold. At the highest irradiance studied, on the other hand, reduced efficiency of photosensitization is attributed to increased photodegradation of doxycycline from higher excited states by processes such as photoionization. A model consistent with these observations is presented along with calculations, based on simple rate equations, that fit the essentials of the proposed model.

Mechanistic investigation of doxycycline photosensitization by picosecond-pulsed and continuous wave laser irradiation of cells in culture. In order to elucidate the photophysical mechanisms of cellular phototoxicity sensitized by doxycycline, MGH-U1 human bladder carcinoma cells in vitro were treated with 20.7 microM doxycycline and irradiated with either a pulsed (lambda = 355 nm, pulse duration = 24 ps) or a continuous wave (lambda = 351 nm) laser. Cumulative radiant exposure and irradiance were systematically varied in experiments with both lasers. Phototoxicity was assessed by epifluorescence microscopy of unfixed cells using rhodamine 123 labeling of mitochondria. With the continuous wave source, the cumulative radiant exposure required for induction of phototoxic injury was independent of irradiance. With the 24-ps-pulsed source, a significantly lower cumulative radiant exposure was required to induce the phototoxicity when the peak irradiance was 5.8 x 10(7) or 1.3 x 10(8) watts cm-2 compared with when peak irradiance was either lower (6.0 x 10(6) watts cm-2) or higher (7.6 x 10(8) watts cm-2). The measured fluorescence lifetimes of doxycycline in buffered saline solution were longer than the laser pulse duration of 24 ps. The increased efficiency of photosensitization at the optimal peak irradiance in the ps domain appears to result from sequential multiphoton absorption involving higher excited states of the singlet manifold. At the highest irradiance studied, on the other hand, reduced efficiency of photosensitization is attributed to increased photodegradation of doxycycline from higher excited states by processes such as photoionization. A model consistent with these observations is presented along with calculations, based on simple rate equations, that fit the essentials of the proposed model.