Datasets:

mjkmain

/

pubmed-subset-1M

like 0

pubmed23n0063_6137

Moment analysis of a multibreath nitrogen washout based on an alveolar gas dilution number.

A common method for analyzing a multibreath nitrogen washout (MBNW) is to perform moment analysis and derive the mean dilution number (MDN). A homogeneously mixed alveolar space with zero series dead space (VD = 0) will always result in a MDN = 1, regardless of breathing pattern. A higher MDN implies more inhomogeneity. But, if VD greater than 0, the MDN can become sensitive (artificially high) to VD/VT ratios. We present an alveolar-based mean dilution number (AMDN) that uses the cumulative expired alveolar volume. Unlike the MDN, the AMDN for a homogeneously mixed alveolar space is unity, regardless of VD or VT, and hence should be a more appropriate index of inhomogeneity at the alveolar level. Two sets of experiments were used to compare the AMDN with the MDN. First, a MBNW was performed by five healthy subjects at spontaneous VD/VT and at a low VD/VT achieved by a controlled increase in VT. Here, the MDN decreased from 1.98 +/- 0.1 to 1.79 +/- 0.06, whereas the AMDN was essentially unchanged (1.42 +/- 0.04 to 1.38 +/- 0.06). Second, MBNW values from seven healthy subjects, five with cystic fibrosis, and 10 asthmatic subjects (before and after bronchodilation) were analyzed. Compared with the MDN, the AMDN showed a significantly wider separation between clinical groups. Also, the AMDN demonstrated an increased variability within both sick groups versus a decrease in the healthy group. We conclude that the AMDN is superior to the MDN because of its decreased sensitivity to breathing pattern but increased sensitivity to degree of disease.

Moment analysis of a multibreath nitrogen washout based on an alveolar gas dilution number. A common method for analyzing a multibreath nitrogen washout (MBNW) is to perform moment analysis and derive the mean dilution number (MDN). A homogeneously mixed alveolar space with zero series dead space (VD = 0) will always result in a MDN = 1, regardless of breathing pattern. A higher MDN implies more inhomogeneity. But, if VD greater than 0, the MDN can become sensitive (artificially high) to VD/VT ratios. We present an alveolar-based mean dilution number (AMDN) that uses the cumulative expired alveolar volume. Unlike the MDN, the AMDN for a homogeneously mixed alveolar space is unity, regardless of VD or VT, and hence should be a more appropriate index of inhomogeneity at the alveolar level. Two sets of experiments were used to compare the AMDN with the MDN. First, a MBNW was performed by five healthy subjects at spontaneous VD/VT and at a low VD/VT achieved by a controlled increase in VT. Here, the MDN decreased from 1.98 +/- 0.1 to 1.79 +/- 0.06, whereas the AMDN was essentially unchanged (1.42 +/- 0.04 to 1.38 +/- 0.06). Second, MBNW values from seven healthy subjects, five with cystic fibrosis, and 10 asthmatic subjects (before and after bronchodilation) were analyzed. Compared with the MDN, the AMDN showed a significantly wider separation between clinical groups. Also, the AMDN demonstrated an increased variability within both sick groups versus a decrease in the healthy group. We conclude that the AMDN is superior to the MDN because of its decreased sensitivity to breathing pattern but increased sensitivity to degree of disease.

pubmed23n0603_11176

Yeast Barcoders: a chemogenomic application of a universal donor-strain collection carrying bar-code identifiers.

The ability to perform complex bioassays in parallel enables experiments that are otherwise impossible because of throughput and cost constraints. For example, highly parallel chemical-genetic screens using pooled collections of thousands of defined Saccharomyces cerevisiae gene deletion strains are feasible because each strain is bar-coded with unique DNA sequences. It is, however, time-consuming and expensive to individually bar-code individual strains. To provide a simple and general method of barcoding yeast collections, we built a set of donor strains, called Barcoders, with unique bar codes that can be systematically transferred to any S. cerevisiae collection. We applied this technology by generating a collection of bar-coded 'decreased abundance by mRNA perturbation' (DAmP) loss-of-function strains comprising 87.1% of all essential yeast genes. These experiments validate both the Barcoders and the DAmP strain collection as useful tools for genome-wide chemical-genetic assays.

Yeast Barcoders: a chemogenomic application of a universal donor-strain collection carrying bar-code identifiers. The ability to perform complex bioassays in parallel enables experiments that are otherwise impossible because of throughput and cost constraints. For example, highly parallel chemical-genetic screens using pooled collections of thousands of defined Saccharomyces cerevisiae gene deletion strains are feasible because each strain is bar-coded with unique DNA sequences. It is, however, time-consuming and expensive to individually bar-code individual strains. To provide a simple and general method of barcoding yeast collections, we built a set of donor strains, called Barcoders, with unique bar codes that can be systematically transferred to any S. cerevisiae collection. We applied this technology by generating a collection of bar-coded 'decreased abundance by mRNA perturbation' (DAmP) loss-of-function strains comprising 87.1% of all essential yeast genes. These experiments validate both the Barcoders and the DAmP strain collection as useful tools for genome-wide chemical-genetic assays.

pubmed23n0765_23333

Novel mutations and expression alterations in SMAD3/TGFBR2 genes in oral carcinoma correlate with poor prognosis.

Transforming growth factor beta (TGF-β) signaling is a pleiotropic cytokine signaling pathway, which controls cellular activities ranging from embryogenesis to apoptosis. Although many molecular alterations in this pathway have been described in cancers, the central point of concern, that is how these alterations influence the treatment outcome, has been addressed to a lesser extent. In this study, we have characterized the alterations of TGF-β-SMAD signaling in 97 oral squamous cell carcinoma (OSCC) samples and assessed the association between these alterations and the outcome of the treatment. Genomic level alteration analysis using reverse transcriptase polymerase chain reaction-single-strand conformation polymorphism/sequencing revealed that there were 25% samples harboring genomic level alterations in this pathway. Altogether, 21% samples showed TGFBR2 mutations, whereas three cases were found to harbor novel SMAD3 mutations. Notably, 14 out of 24 TGFBR2 mutations are of one type (c.*6C>A), which supplemented complementarity for hsa-miR-3189-5p. These samples showed significantly low TGFBR2 transcript levels (P = 0.026). In addition, transcript level studies using quantitative real-time PCR revealed a strong association between low TGFBR2 transcript levels and poor disease-free survival (P = 0.028) as well as poor overall survival (P = 0.013). In brief, our results showed that oral cancers with TGFBR2 downregulation comprise a different group with more aggressive nature. These results suggest that in OSCCs, TGFBR2 transcript levels may be developed as a promising prognostic biomarker. Furthermore, for the first time, this study reports SMAD3 mutations in oral carcinoma.

Novel mutations and expression alterations in SMAD3/TGFBR2 genes in oral carcinoma correlate with poor prognosis. Transforming growth factor beta (TGF-β) signaling is a pleiotropic cytokine signaling pathway, which controls cellular activities ranging from embryogenesis to apoptosis. Although many molecular alterations in this pathway have been described in cancers, the central point of concern, that is how these alterations influence the treatment outcome, has been addressed to a lesser extent. In this study, we have characterized the alterations of TGF-β-SMAD signaling in 97 oral squamous cell carcinoma (OSCC) samples and assessed the association between these alterations and the outcome of the treatment. Genomic level alteration analysis using reverse transcriptase polymerase chain reaction-single-strand conformation polymorphism/sequencing revealed that there were 25% samples harboring genomic level alterations in this pathway. Altogether, 21% samples showed TGFBR2 mutations, whereas three cases were found to harbor novel SMAD3 mutations. Notably, 14 out of 24 TGFBR2 mutations are of one type (c.*6C>A), which supplemented complementarity for hsa-miR-3189-5p. These samples showed significantly low TGFBR2 transcript levels (P = 0.026). In addition, transcript level studies using quantitative real-time PCR revealed a strong association between low TGFBR2 transcript levels and poor disease-free survival (P = 0.028) as well as poor overall survival (P = 0.013). In brief, our results showed that oral cancers with TGFBR2 downregulation comprise a different group with more aggressive nature. These results suggest that in OSCCs, TGFBR2 transcript levels may be developed as a promising prognostic biomarker. Furthermore, for the first time, this study reports SMAD3 mutations in oral carcinoma.

pubmed23n0904_20543

Fluvastatin inhibits intimal hyperplasia in wild-type but not Thbs1-null mice.

Thrombospondin-1 (TSP-1) is functionally important to intimal hyperplasia (IH) development. Statin drugs have beneficial pleiotropic effects, including reduced IH; however, the effect of statins on IH in a TSP-1-independent setting is unknown. Statins will be less effective in attenuating IH after vascular injury in TSP-1-null (Thbs1<sup-/-</sup) mice compared with wild-type (WT) mice. Carotid artery ligation was performed on WT and Thbs1<sup-/-</sup mice. Each strain was divided into two groups: no statin control or standard chow containing fluvastatin (10 or 40 mg/kg/d). After 28 d, analysis included morphometric analysis and real-time quantitative reverse transcription polymerase chain reaction on the arteries and enzyme-linked immunosorbent assay on plasma (TSP-1 WT, TSP-2 WT, and Thbs1<sup-/-</sup). Comparisons were made by analysis of variance, with P &lt; 0.05 considered significant. In no statin controls, WT mice had more IH than Thbs1<sup-/-</sup mice (0.46 ± 0.09 versus 0.15 ± 0.04). Fluvastatin reduced IH in the WT (0.46 ± 0.09 versus 0.23 ± 0.06), but not in Thbs1<sup-/-</sup groups (0.15 ± 0.04 versus 0.22 ± 0.07). No difference in IH existed between Thbs1<sup-/-</sup no statin controls and fluvastatin WT and Thbs1<sup-/-</sup groups. Statin dose did not affect IH. TSP-1 plasma levels were increased in fluvastatin WT. TSP-2 levels were decreased in fluvastatin WT and elevated in fluvastatin Thbs1<sup-/-</sup. Fluvastatin had no effect on tissue Thbs1 or Thbs2 gene expression. TSP-1 is necessary for robust IH after arterial injury. Because fluvastatin had no effect on IH in Thbs1<sup-/-</sup, the data suggest that the statin effect on IH may be largely TSP-1 dependent. Both statins and the presence of TSP-1 affect TSP-1 and TSP-2 plasma levels.

Fluvastatin inhibits intimal hyperplasia in wild-type but not Thbs1-null mice. Thrombospondin-1 (TSP-1) is functionally important to intimal hyperplasia (IH) development. Statin drugs have beneficial pleiotropic effects, including reduced IH; however, the effect of statins on IH in a TSP-1-independent setting is unknown. Statins will be less effective in attenuating IH after vascular injury in TSP-1-null (Thbs1<sup-/-</sup) mice compared with wild-type (WT) mice. Carotid artery ligation was performed on WT and Thbs1<sup-/-</sup mice. Each strain was divided into two groups: no statin control or standard chow containing fluvastatin (10 or 40 mg/kg/d). After 28 d, analysis included morphometric analysis and real-time quantitative reverse transcription polymerase chain reaction on the arteries and enzyme-linked immunosorbent assay on plasma (TSP-1 WT, TSP-2 WT, and Thbs1<sup-/-</sup). Comparisons were made by analysis of variance, with P &lt; 0.05 considered significant. In no statin controls, WT mice had more IH than Thbs1<sup-/-</sup mice (0.46 ± 0.09 versus 0.15 ± 0.04). Fluvastatin reduced IH in the WT (0.46 ± 0.09 versus 0.23 ± 0.06), but not in Thbs1<sup-/-</sup groups (0.15 ± 0.04 versus 0.22 ± 0.07). No difference in IH existed between Thbs1<sup-/-</sup no statin controls and fluvastatin WT and Thbs1<sup-/-</sup groups. Statin dose did not affect IH. TSP-1 plasma levels were increased in fluvastatin WT. TSP-2 levels were decreased in fluvastatin WT and elevated in fluvastatin Thbs1<sup-/-</sup. Fluvastatin had no effect on tissue Thbs1 or Thbs2 gene expression. TSP-1 is necessary for robust IH after arterial injury. Because fluvastatin had no effect on IH in Thbs1<sup-/-</sup, the data suggest that the statin effect on IH may be largely TSP-1 dependent. Both statins and the presence of TSP-1 affect TSP-1 and TSP-2 plasma levels.

pubmed23n0935_13793

Wanted: Expert operators for coronary chronic total occlusion interventions.

Coronary chronic total occlusions can be recanalized by expert operators with high success and acceptable complication rates. Developing a procedural plan (using the hybrid or another algorithm) is critical for success. Live case demonstrations appear safe and can be of high educational value. The key factor for achieving good outcomes in chronic total occlusion interventions remains operator expertise. Development of more expert operators will improve the treatment options for this challenging group of lesions and patients.

Wanted: Expert operators for coronary chronic total occlusion interventions. Coronary chronic total occlusions can be recanalized by expert operators with high success and acceptable complication rates. Developing a procedural plan (using the hybrid or another algorithm) is critical for success. Live case demonstrations appear safe and can be of high educational value. The key factor for achieving good outcomes in chronic total occlusion interventions remains operator expertise. Development of more expert operators will improve the treatment options for this challenging group of lesions and patients.

pubmed23n0894_3886

Potential prebiotic properties of cashew apple (Anacardium occidentale L.) agro-industrial byproduct on Lactobacillus species.

The prebiotic effects of a cashew apple (Anacardium occidentale L.) agro-industrial byproduct powder (CAP) on different potentially probiotic Lactobacillus strains, namely Lactobacillus acidophilus LA-05, Lactobacillus casei L-26 and Lactobacillus paracasei L-10, were assessed using in vitro experimental models. Accordingly, the growth of the Lactobacillus strains when cultivated in a broth containing CAP (20 or 30 g L<sup-1</sup ), glucose (20 g L<sup-1</sup ) or fructooligosaccharides (FOS) (20 g L<sup-1</sup ) was monitored over 48 h; the prebiotic activity scores of CAP were determined; and the changes in pH values, production of organic acids and consumption of sugars in growth media were verified. During the 48-h cultivation, similar viable cell counts were observed for the Lactobacillus strains grown in the different media tested. The CAP presented positive prebiotic activity scores toward all the tested Lactobacillus strains, indicating a desirable selective fermentable activity relative to enteric organisms. The cultivation of the Lactobacillus strains in broth containing glucose, FOS or CAP resulted in high viable cell counts, a decreased pH, the production of organic acids and the consumption of sugars over time, revealing intense bacterial metabolic activity. The CAP exerts potential prebiotic effects on different potentially probiotic Lactobacillus strains and should be an added-value ingredient for the food industry. © 2017 Society of Chemical Industry.

Potential prebiotic properties of cashew apple (Anacardium occidentale L.) agro-industrial byproduct on Lactobacillus species. The prebiotic effects of a cashew apple (Anacardium occidentale L.) agro-industrial byproduct powder (CAP) on different potentially probiotic Lactobacillus strains, namely Lactobacillus acidophilus LA-05, Lactobacillus casei L-26 and Lactobacillus paracasei L-10, were assessed using in vitro experimental models. Accordingly, the growth of the Lactobacillus strains when cultivated in a broth containing CAP (20 or 30 g L<sup-1</sup ), glucose (20 g L<sup-1</sup ) or fructooligosaccharides (FOS) (20 g L<sup-1</sup ) was monitored over 48 h; the prebiotic activity scores of CAP were determined; and the changes in pH values, production of organic acids and consumption of sugars in growth media were verified. During the 48-h cultivation, similar viable cell counts were observed for the Lactobacillus strains grown in the different media tested. The CAP presented positive prebiotic activity scores toward all the tested Lactobacillus strains, indicating a desirable selective fermentable activity relative to enteric organisms. The cultivation of the Lactobacillus strains in broth containing glucose, FOS or CAP resulted in high viable cell counts, a decreased pH, the production of organic acids and the consumption of sugars over time, revealing intense bacterial metabolic activity. The CAP exerts potential prebiotic effects on different potentially probiotic Lactobacillus strains and should be an added-value ingredient for the food industry. © 2017 Society of Chemical Industry.

pubmed23n1095_11064

Chronic Thromboembolic Pulmonary Hypertension Secondary to Thrombophilia and Incidentally Diagnosed Atrial Septal Defect.

A 46-year-old man developed chronic thromboembolic pulmonary hypertension and atrial fibrillation after acute pulmonary embolism. He was found incidentally to have an isolated secundum atrial septal defect, as well as a homozygous mutation for the plasminogen activator inhibitor-1 gene. He was successfully treated with pulmonary endarterectomy and atrial septal defect repair. He has continued to do well on a regimen of dabigatran. (<bLevel of Difficulty: Beginner.</b).

Chronic Thromboembolic Pulmonary Hypertension Secondary to Thrombophilia and Incidentally Diagnosed Atrial Septal Defect. A 46-year-old man developed chronic thromboembolic pulmonary hypertension and atrial fibrillation after acute pulmonary embolism. He was found incidentally to have an isolated secundum atrial septal defect, as well as a homozygous mutation for the plasminogen activator inhibitor-1 gene. He was successfully treated with pulmonary endarterectomy and atrial septal defect repair. He has continued to do well on a regimen of dabigatran. (<bLevel of Difficulty: Beginner.</b).

pubmed23n1028_23953

Adsorption-induced co-assembly of hairy and isotropic particles.

We use coarse-grained molecular dynamics simulations to study the behavior of polymer-tethered particles immersed in fluids of isotropic particles. Particles modified with weakly anchored, mobile ligands are considered. We discuss how the concentration of fluid particles affects the morphology of an isolated hairy particle. It is shown that hairy particles present different morphologies including typical core-shell, octopus-like and corn-like, depending on fluid-segment interactions and the fluid density. The mechanism of changes in the shape of hairy particles is explained. The reconfiguration of the polymer corona arises from adsorption of fluid particles "on chains". The adsorbed fluid particles form bridges between the chains. This causes the mobile ligands to merge into clusters on the core surface. A part of the core remains empty so the hairy particle becomes a Janus-like object. We also study co-assembly in mixtures of hairy and isotropic particles. Depending on the strength of fluid-segment interactions, hairy particles with fluid particles trapped inside their coronas remain isolated or form mixed clusters of different structures. The aggregation of hairy particles results from the formation of bridges between chains belonging to different cores by fluid particles.

Adsorption-induced co-assembly of hairy and isotropic particles. We use coarse-grained molecular dynamics simulations to study the behavior of polymer-tethered particles immersed in fluids of isotropic particles. Particles modified with weakly anchored, mobile ligands are considered. We discuss how the concentration of fluid particles affects the morphology of an isolated hairy particle. It is shown that hairy particles present different morphologies including typical core-shell, octopus-like and corn-like, depending on fluid-segment interactions and the fluid density. The mechanism of changes in the shape of hairy particles is explained. The reconfiguration of the polymer corona arises from adsorption of fluid particles "on chains". The adsorbed fluid particles form bridges between the chains. This causes the mobile ligands to merge into clusters on the core surface. A part of the core remains empty so the hairy particle becomes a Janus-like object. We also study co-assembly in mixtures of hairy and isotropic particles. Depending on the strength of fluid-segment interactions, hairy particles with fluid particles trapped inside their coronas remain isolated or form mixed clusters of different structures. The aggregation of hairy particles results from the formation of bridges between chains belonging to different cores by fluid particles.

pubmed23n1157_13234

Acute kidney injury in the tropics.

The tropics are a region consisting of more than 125 countries, accounting for 40% of the world's population. The region's population is expected to increase up to 60% in the coming decades. Many tropical countries continue to experience public health problems such as high rates of infectious diseases, lack of sanitation, climate change impacts, poor regulation of herbal medicines and low access to healthcare. These conditions produce the unique problem of tropical acute kidney injury (AKI), which is associated with high morbidity and mortality. Tropical infections such as leptospirosis, dengue and malaria have varied mechanisms of AKI, including both direct kidney invasion and indirect effects, depending on the disease characteristics. Animal toxins from snakebites and arthropods along with plant toxins, such as djenkol beans, starfruit and herbal medicine, are characterized by a harmful renal effect from each toxic substance. Environmental factors such as heat stress, natural disasters and chemical compounds also lead to AKI and have a systemic effect from their own pathogenesis. The long-term kidney prognosis varies among these etiologies depending on the cause and severity of disease. However, all these conditions are potentially preventable and treatable. Prompt management and good preventive approaches are needed. This article will focus on the epidemiology, pathogenesis and management of AKI associated with tropical infections, toxins and environment impacts.

Acute kidney injury in the tropics. The tropics are a region consisting of more than 125 countries, accounting for 40% of the world's population. The region's population is expected to increase up to 60% in the coming decades. Many tropical countries continue to experience public health problems such as high rates of infectious diseases, lack of sanitation, climate change impacts, poor regulation of herbal medicines and low access to healthcare. These conditions produce the unique problem of tropical acute kidney injury (AKI), which is associated with high morbidity and mortality. Tropical infections such as leptospirosis, dengue and malaria have varied mechanisms of AKI, including both direct kidney invasion and indirect effects, depending on the disease characteristics. Animal toxins from snakebites and arthropods along with plant toxins, such as djenkol beans, starfruit and herbal medicine, are characterized by a harmful renal effect from each toxic substance. Environmental factors such as heat stress, natural disasters and chemical compounds also lead to AKI and have a systemic effect from their own pathogenesis. The long-term kidney prognosis varies among these etiologies depending on the cause and severity of disease. However, all these conditions are potentially preventable and treatable. Prompt management and good preventive approaches are needed. This article will focus on the epidemiology, pathogenesis and management of AKI associated with tropical infections, toxins and environment impacts.

pubmed23n0581_1486

Preparation and biodegradation of sugar-containing poly(vinyl acetate) emulsions.

To accelerate the biodegradability of poly(vinyl acetate)-based emulsions, emulsion copolymerizations of vinyl sugars, including triacetylated N-acetyl-D-glucosamine (GlcNAc)-substituted 2-hydroxyethyl methacrylate (GlcNAc(Ac)3-substituted HEMA), glucose-substituted HEMA (GEMA) and 6-O-vinyladipoyl-D-glucose (6-O-VAG) with vinyl acetate (VAc), were carried out using poly(vinyl alcohol) as an emulsifying agent in the presence of poly[(butylene succinate)-co-(butylene adipate)] [poly(BS-co-BA)]. Copolymerization with GEMA produced a stable emulsion and that with 6-O-VAG also produced a homogeneous emulsion. Their biodegradation tests indicated that PVAc main chain scission was accelerated by copolymerization with vinyl sugars.

Preparation and biodegradation of sugar-containing poly(vinyl acetate) emulsions. To accelerate the biodegradability of poly(vinyl acetate)-based emulsions, emulsion copolymerizations of vinyl sugars, including triacetylated N-acetyl-D-glucosamine (GlcNAc)-substituted 2-hydroxyethyl methacrylate (GlcNAc(Ac)3-substituted HEMA), glucose-substituted HEMA (GEMA) and 6-O-vinyladipoyl-D-glucose (6-O-VAG) with vinyl acetate (VAc), were carried out using poly(vinyl alcohol) as an emulsifying agent in the presence of poly[(butylene succinate)-co-(butylene adipate)] [poly(BS-co-BA)]. Copolymerization with GEMA produced a stable emulsion and that with 6-O-VAG also produced a homogeneous emulsion. Their biodegradation tests indicated that PVAc main chain scission was accelerated by copolymerization with vinyl sugars.

pubmed23n0617_13398

Using empirical phase diagrams to understand the role of intramolecular dynamics in immunoglobulin G stability.

Understanding the relationship between protein dynamics and stability is of paramount importance to the fields of biology and pharmaceutics. Clarifying this relationship is complicated by the large amount of experimental data that must be generated and analyzed if motions that exist over the wide range of timescales are to be included. To address this issue, we propose an approach that utilizes a multidimensional vector-based empirical phase diagram (EPD) to analyze a set of dynamic results acquired across a temperature-pH perturbation plane. This approach is applied to a humanized immunoglobulin G1 (IgG1), a protein of major biological and pharmaceutical importance whose dynamic nature is linked to its multiple biological roles. Static and dynamic measurements are used to characterize the IgG and to construct both static and dynamic EPDs. Between pH 5 and 8, a single, pH-dependent transition is observed that corresponds to thermal unfolding of the IgG. Under more acidic conditions, evidence exists for the formation of a more compact, aggregation resistant state of the immunoglobulin, known as A-form. The dynamics-based EPD presents a considerably more detailed pattern of apparent phase transitions over the temperature-pH plane. The utility and potential applications of this approach are discussed.

Using empirical phase diagrams to understand the role of intramolecular dynamics in immunoglobulin G stability. Understanding the relationship between protein dynamics and stability is of paramount importance to the fields of biology and pharmaceutics. Clarifying this relationship is complicated by the large amount of experimental data that must be generated and analyzed if motions that exist over the wide range of timescales are to be included. To address this issue, we propose an approach that utilizes a multidimensional vector-based empirical phase diagram (EPD) to analyze a set of dynamic results acquired across a temperature-pH perturbation plane. This approach is applied to a humanized immunoglobulin G1 (IgG1), a protein of major biological and pharmaceutical importance whose dynamic nature is linked to its multiple biological roles. Static and dynamic measurements are used to characterize the IgG and to construct both static and dynamic EPDs. Between pH 5 and 8, a single, pH-dependent transition is observed that corresponds to thermal unfolding of the IgG. Under more acidic conditions, evidence exists for the formation of a more compact, aggregation resistant state of the immunoglobulin, known as A-form. The dynamics-based EPD presents a considerably more detailed pattern of apparent phase transitions over the temperature-pH plane. The utility and potential applications of this approach are discussed.

pubmed23n0770_4669

Understanding Washington: a nephrologist's perspective from inside the Beltway.

The major principles that drive U.S. federal health policy-making are: (1) fixed or reduced costs, (2) ensured outcomes (or no evidence of undertreatment), (3) streamlined administration, and (4) political viability. A corollary is that providers are uniquely sensitive to financial incentives. Understanding these principles is vital to understanding federal health policy. Critically, these principles are nonpartisan and have been supported and used by all administrations since President Reagan. This article examines the end-stage renal disease (ESRD) prospective payment system, colloquially called "The Bundle," in the context of these major principles. Successful health policy, successful legislation, and successful regulation building all require executive leadership, mutual trust, and compromise. This is demonstrated by the events surrounding the passage of the Medicare inpatient prospective payment system, which governs hospital reimbursement for Medicare beneficiaries, including those not covered in the ESRD program. Given that the ESRD benefit consumes 6.3% of the Medicare budget for approximately 2% of Medicare beneficiaries, if nephrology is to experience future success, we must change how both policymakers and the wider field of medicine perceive our specialty. Understanding the major principles behind health care policy may facilitate this goal.

Understanding Washington: a nephrologist's perspective from inside the Beltway. The major principles that drive U.S. federal health policy-making are: (1) fixed or reduced costs, (2) ensured outcomes (or no evidence of undertreatment), (3) streamlined administration, and (4) political viability. A corollary is that providers are uniquely sensitive to financial incentives. Understanding these principles is vital to understanding federal health policy. Critically, these principles are nonpartisan and have been supported and used by all administrations since President Reagan. This article examines the end-stage renal disease (ESRD) prospective payment system, colloquially called "The Bundle," in the context of these major principles. Successful health policy, successful legislation, and successful regulation building all require executive leadership, mutual trust, and compromise. This is demonstrated by the events surrounding the passage of the Medicare inpatient prospective payment system, which governs hospital reimbursement for Medicare beneficiaries, including those not covered in the ESRD program. Given that the ESRD benefit consumes 6.3% of the Medicare budget for approximately 2% of Medicare beneficiaries, if nephrology is to experience future success, we must change how both policymakers and the wider field of medicine perceive our specialty. Understanding the major principles behind health care policy may facilitate this goal.

pubmed23n0754_6060

Effect of postmortem interval on the graft endothelium during preservation and after transplantation for keratoconus.

To investigate the effect of postmortem intervals and prognostic factors on endothelial cell density (ECD) of human donor corneas during preservation and at 1 and 3 years after transplantation in patients transplanted for keratoconus. Two different studies were performed: (1) with 733 donor corneas selected for the preservation study and (2) 64 patients with keratoconus selected retrospectively from 2 hospital clinics. The corneas were evaluated on the basis of the ECD during preservation, study A, and at 1 and 3 years after transplantation, study B. The effect of ≥ 10 hours of postmortem interval on the percentage of corneal endothelial cell loss (ECL) was determined. The multiple regression showed no statistical significance (P = 0.827) of postmortem interval on ECL during preservation. However, for patients with keratoconus, the postmortem interval was statistically significant at both 1 year (P &lt; 0.0001) and 3 years after transplantation (P &lt; 0.0001). The postmortem interval has no influence on the ECD during preservation. However, it has a statistically significant effect on the ECL after transplantation for patients transplanted for keratoconus, and therefore, it becomes eligible to be one of the potential factors affecting the ECD apart from surgical trauma.

Effect of postmortem interval on the graft endothelium during preservation and after transplantation for keratoconus. To investigate the effect of postmortem intervals and prognostic factors on endothelial cell density (ECD) of human donor corneas during preservation and at 1 and 3 years after transplantation in patients transplanted for keratoconus. Two different studies were performed: (1) with 733 donor corneas selected for the preservation study and (2) 64 patients with keratoconus selected retrospectively from 2 hospital clinics. The corneas were evaluated on the basis of the ECD during preservation, study A, and at 1 and 3 years after transplantation, study B. The effect of ≥ 10 hours of postmortem interval on the percentage of corneal endothelial cell loss (ECL) was determined. The multiple regression showed no statistical significance (P = 0.827) of postmortem interval on ECL during preservation. However, for patients with keratoconus, the postmortem interval was statistically significant at both 1 year (P &lt; 0.0001) and 3 years after transplantation (P &lt; 0.0001). The postmortem interval has no influence on the ECD during preservation. However, it has a statistically significant effect on the ECL after transplantation for patients transplanted for keratoconus, and therefore, it becomes eligible to be one of the potential factors affecting the ECD apart from surgical trauma.

pubmed23n0765_8211

Water in the hydration shell of halide ions has significantly reduced Fermi resonance and moderately enhanced Raman cross section in the OH stretch regions.

Water in the presence of electrolytes plays an important role in biological and industrial processes. The properties of water, such as the intermolecular coupling, Fermi resonance (FR), hydrogen-bonding, and Raman cross section were investigated by measuring the Raman spectra in the OD and OH stretch regions in presence of alkali halides (NaX; X = F, Cl, Br, I). It is observed that the changes in spectral characteristics by the addition of NaX in D2O are similar to those obtained by the addition of H2O in D2O. The spectral width decreases significantly by the addition of NaX in D2O (H2O) than that in the isotopically diluted water. Quantitative estimation, on the basis of integrated Raman intensity, revealed that the relative Raman cross section, σ(H)/σ(b) (σ(H) and σ(b) are the average Raman cross section of water in the first hydration shell of X(-) and in bulk, respectively), in D2O and H2O is higher than those in the respective isotopically diluted water. These results suggest that water in the hydration shell has reduced FR and intermolecular coupling compared to those in bulk. In the isotopically diluted water, the relative Raman cross section increases with increase in size of the halide ions (σ(H)/σ(b) = 0.6, 1.1, 1.5, and 1.9 for F(-), Cl(-), Br(-), and I(-), respectively), which is assignable to the enhancement of Raman cross section by charge transfer from halide ions to the hydrating water. Nevertheless, the experimentally determined σ(H)/σ(b) is lower than the calculated values obtained on the basis of the energy of the charge transfer state of water. The weak enhancement of σ(H)/σ(b) signifies that the charge transfer transition in the hydration shell of halide ions causes little change in the OD (OH) bond lengths of hydrating water.

Water in the hydration shell of halide ions has significantly reduced Fermi resonance and moderately enhanced Raman cross section in the OH stretch regions. Water in the presence of electrolytes plays an important role in biological and industrial processes. The properties of water, such as the intermolecular coupling, Fermi resonance (FR), hydrogen-bonding, and Raman cross section were investigated by measuring the Raman spectra in the OD and OH stretch regions in presence of alkali halides (NaX; X = F, Cl, Br, I). It is observed that the changes in spectral characteristics by the addition of NaX in D2O are similar to those obtained by the addition of H2O in D2O. The spectral width decreases significantly by the addition of NaX in D2O (H2O) than that in the isotopically diluted water. Quantitative estimation, on the basis of integrated Raman intensity, revealed that the relative Raman cross section, σ(H)/σ(b) (σ(H) and σ(b) are the average Raman cross section of water in the first hydration shell of X(-) and in bulk, respectively), in D2O and H2O is higher than those in the respective isotopically diluted water. These results suggest that water in the hydration shell has reduced FR and intermolecular coupling compared to those in bulk. In the isotopically diluted water, the relative Raman cross section increases with increase in size of the halide ions (σ(H)/σ(b) = 0.6, 1.1, 1.5, and 1.9 for F(-), Cl(-), Br(-), and I(-), respectively), which is assignable to the enhancement of Raman cross section by charge transfer from halide ions to the hydrating water. Nevertheless, the experimentally determined σ(H)/σ(b) is lower than the calculated values obtained on the basis of the energy of the charge transfer state of water. The weak enhancement of σ(H)/σ(b) signifies that the charge transfer transition in the hydration shell of halide ions causes little change in the OD (OH) bond lengths of hydrating water.

pubmed23n1030_15764

Mast Cells and Blood Vessels Profile in Oral Carcinogenesis: An Immunohistochemistry Study.

The objectives of the present study were to evaluate angiogenesis and mast cell density in oral epithelial dysplasia and oral squamous cell carcinoma (OSCC). This was an observational, retrospective and quantitative study. The samples consisted of 60 tissue specimens from patients with squamous cell carcinoma, epithelial dysplasia and controls (n=20/group). Immunohistochemistry was performed using an anti-tryptase antibody to mast cells and anti-CD31 and anti-CD34 for blood vessels and we count the number of mast cells and determine the percentage of CD31 and CD34 antibody staining (vascular density). The mast cells had lower density in OSCC compared to control and dysplasia (p = 0.009). In angiogenesis, the expression of CD31 showed a higher percentage of blood vessels in OSCC (p &lt; 0.001), however, CD34 showed no difference between groups (p=0.092). The CD31 antibody presented as a high immunostaining in oral mucosa than CD34. The increased vascularity in squamous cell carcinoma suggests that angiogenesis begins when malignant transformation starts that seems to be inversely associated with the number of mast cells.

Mast Cells and Blood Vessels Profile in Oral Carcinogenesis: An Immunohistochemistry Study. The objectives of the present study were to evaluate angiogenesis and mast cell density in oral epithelial dysplasia and oral squamous cell carcinoma (OSCC). This was an observational, retrospective and quantitative study. The samples consisted of 60 tissue specimens from patients with squamous cell carcinoma, epithelial dysplasia and controls (n=20/group). Immunohistochemistry was performed using an anti-tryptase antibody to mast cells and anti-CD31 and anti-CD34 for blood vessels and we count the number of mast cells and determine the percentage of CD31 and CD34 antibody staining (vascular density). The mast cells had lower density in OSCC compared to control and dysplasia (p = 0.009). In angiogenesis, the expression of CD31 showed a higher percentage of blood vessels in OSCC (p &lt; 0.001), however, CD34 showed no difference between groups (p=0.092). The CD31 antibody presented as a high immunostaining in oral mucosa than CD34. The increased vascularity in squamous cell carcinoma suggests that angiogenesis begins when malignant transformation starts that seems to be inversely associated with the number of mast cells.

pubmed23n0490_65

[Zinc supplementation may recover taste for salt meals].

To evaluate the effect of zinc on the appetite for salt foods in children aged 8 months to 5 years. Double-blind, placebo-controlled study. Two groups of 20 children refusing to eat salt foods were followed during 6 months. The children in the first group received zinc chelate 1 mg/kg daily for 3 months. The second group received a placebo solution. The two groups were similar in terms of age, sex, weight, duration of breastfeeding, age at weaning, biochemical and hematological data. The response of children to treatment was informed by their mothers. 17/20 (85%) of the children receiving zinc chelate and 10/20 (50%) of the children receiving placebo improved their appetite for salt foods. The difference was statistically significant (p &lt; 0.05, chi-square test). Zinc supplementation may improve the acceptance of salt foods by children.

[Zinc supplementation may recover taste for salt meals]. To evaluate the effect of zinc on the appetite for salt foods in children aged 8 months to 5 years. Double-blind, placebo-controlled study. Two groups of 20 children refusing to eat salt foods were followed during 6 months. The children in the first group received zinc chelate 1 mg/kg daily for 3 months. The second group received a placebo solution. The two groups were similar in terms of age, sex, weight, duration of breastfeeding, age at weaning, biochemical and hematological data. The response of children to treatment was informed by their mothers. 17/20 (85%) of the children receiving zinc chelate and 10/20 (50%) of the children receiving placebo improved their appetite for salt foods. The difference was statistically significant (p &lt; 0.05, chi-square test). Zinc supplementation may improve the acceptance of salt foods by children.

pubmed23n0411_9947

Trends in the incidence of hepatocellular carcinoma in boys and girls in Taiwan after large-scale hepatitis B vaccination.

In July 1984, large-scale hepatitis B vaccination of newborns began in Taiwan. Vaccination decreased the overall incidence of childhood hepatocellular carcinoma (HCC). We conducted this study to learn whether the vaccination program had the same effect on boys and girls. We collected liver carcinoma (including HCC and hepatoblastoma) deaths from 1974 to 1999 from the Taiwan Mortality Registry and the 1974-1999 population data from the Taiwan Ministry of Interior to calculate the liver carcinoma mortality rate. The populations ages 0-14 and ages 15-100 in each calendar year were treated as the study group and the reference group, respectively. We divided the 1974-1999 calendar years into 4-year strata and calculated the mortality rates of each 4-year period. We used the 1980-1983 mortality rate as the standard to calculate 4-year-interval mortality rate ratios. Vaccination effects by age and gender were estimated dividing the study and the reference groups into male and female subgroups. We used a double-comparison method to confirm the effects of hepatitis B vaccination: the mortality rate trend of the study group (ages 0-14) compared with the reference group (ages 15-100) in the same period (1984-1999), and the mortality rate trend of the study group (age 0-14) compared with itself in the pre- and postvaccination periods (1974-1983, 1984-1999). Liver carcinoma mortality decreased significantly among both males and females after 1984. In the study group, the male mortality rate decreased by up to 70%, and the female mortality rate decreased by up to 62% in the 1996-1999 interval compared with the 1980-1983 period. Both the male and the female study groups' mortality rate trends decreased from 1983 to 1999 compared with the 1974-1983 period or compared with the same period of the reference groups. Our results indicate hepatitis B vaccination decreases childhood HCC in both boys and girls.

Trends in the incidence of hepatocellular carcinoma in boys and girls in Taiwan after large-scale hepatitis B vaccination. In July 1984, large-scale hepatitis B vaccination of newborns began in Taiwan. Vaccination decreased the overall incidence of childhood hepatocellular carcinoma (HCC). We conducted this study to learn whether the vaccination program had the same effect on boys and girls. We collected liver carcinoma (including HCC and hepatoblastoma) deaths from 1974 to 1999 from the Taiwan Mortality Registry and the 1974-1999 population data from the Taiwan Ministry of Interior to calculate the liver carcinoma mortality rate. The populations ages 0-14 and ages 15-100 in each calendar year were treated as the study group and the reference group, respectively. We divided the 1974-1999 calendar years into 4-year strata and calculated the mortality rates of each 4-year period. We used the 1980-1983 mortality rate as the standard to calculate 4-year-interval mortality rate ratios. Vaccination effects by age and gender were estimated dividing the study and the reference groups into male and female subgroups. We used a double-comparison method to confirm the effects of hepatitis B vaccination: the mortality rate trend of the study group (ages 0-14) compared with the reference group (ages 15-100) in the same period (1984-1999), and the mortality rate trend of the study group (age 0-14) compared with itself in the pre- and postvaccination periods (1974-1983, 1984-1999). Liver carcinoma mortality decreased significantly among both males and females after 1984. In the study group, the male mortality rate decreased by up to 70%, and the female mortality rate decreased by up to 62% in the 1996-1999 interval compared with the 1980-1983 period. Both the male and the female study groups' mortality rate trends decreased from 1983 to 1999 compared with the 1974-1983 period or compared with the same period of the reference groups. Our results indicate hepatitis B vaccination decreases childhood HCC in both boys and girls.

pubmed23n0910_15563

Intracellular Pharmacokinetics of Antibacterials and Their Clinical Implications.

The intracellular pharmacokinetics of the different classes of antimicrobials into surrogate markers of tissue accumulation (alveolar macrophages and/or total alveolar cells collected by means of bronchoalveolar lavage or peripheral white blood cells) was reviewed. The aim of this review was to discuss the clinical implications of the intracellular pharmacokinetics of antibacterials, either from the therapeutic or toxicological perspective. The different pharmacokinetic behaviour of antimicrobials within cells is mainly related to their physicochemical properties (hydrophilicity and lipophilicity), and may have several clinical implications. Therapeutic efficacy against intracellular pathogens has been correlated mainly with the intracellular concentrations achieved by the different antimicrobial agents. This is relevant especially for macrolides, tetracyclines, fluoroquinolones and rifampicin in the treatment of bacterial infections such as Legionella pneumophila pneumonia, Mycoplasma pneumoniae pneumonia, non-gonococcal urethritis and chronic staphylococcal infections. Additionally, intracellular accumulation of antibacterials was correlated with the possibility of causing organ-specific toxicity, as in the case of aminoglycosides in regard to the risk of nephrotoxicity. Finally, it should be kept in mind that intracellular accumulation may also represent a drug reservoir in the case of lipophilic antimicrobials. This may become extremely relevant from the clinical standpoint when treating critically ill patients with sepsis with antibacterials. The pathophysiology of sepsis may explain why it is necessary to start therapy with an increased loading dose of hydrophilic antimicrobials to promptly achieve therapeutically effective concentrations.

Intracellular Pharmacokinetics of Antibacterials and Their Clinical Implications. The intracellular pharmacokinetics of the different classes of antimicrobials into surrogate markers of tissue accumulation (alveolar macrophages and/or total alveolar cells collected by means of bronchoalveolar lavage or peripheral white blood cells) was reviewed. The aim of this review was to discuss the clinical implications of the intracellular pharmacokinetics of antibacterials, either from the therapeutic or toxicological perspective. The different pharmacokinetic behaviour of antimicrobials within cells is mainly related to their physicochemical properties (hydrophilicity and lipophilicity), and may have several clinical implications. Therapeutic efficacy against intracellular pathogens has been correlated mainly with the intracellular concentrations achieved by the different antimicrobial agents. This is relevant especially for macrolides, tetracyclines, fluoroquinolones and rifampicin in the treatment of bacterial infections such as Legionella pneumophila pneumonia, Mycoplasma pneumoniae pneumonia, non-gonococcal urethritis and chronic staphylococcal infections. Additionally, intracellular accumulation of antibacterials was correlated with the possibility of causing organ-specific toxicity, as in the case of aminoglycosides in regard to the risk of nephrotoxicity. Finally, it should be kept in mind that intracellular accumulation may also represent a drug reservoir in the case of lipophilic antimicrobials. This may become extremely relevant from the clinical standpoint when treating critically ill patients with sepsis with antibacterials. The pathophysiology of sepsis may explain why it is necessary to start therapy with an increased loading dose of hydrophilic antimicrobials to promptly achieve therapeutically effective concentrations.

pubmed23n0818_19330

New physics in resonant production of Higgs boson pairs.

We advocate a search for an extended scalar sector at the LHC via hh production, where h is the 125 GeV Higgs boson. A resonance feature in the hh invariant mass is a smoking gun of an s-channel heavy Higgs resonance, H. With one h decaying to two photons and the other decaying to b quarks, the resonant signal may be discoverable above the hh continuum background for M(H)&lt;1  TeV. The product of the scalar and top Yukawa couplings can be measured to better than 10%-20% accuracy, and its sign can be inferred from the hh line shape via interference effects.

New physics in resonant production of Higgs boson pairs. We advocate a search for an extended scalar sector at the LHC via hh production, where h is the 125 GeV Higgs boson. A resonance feature in the hh invariant mass is a smoking gun of an s-channel heavy Higgs resonance, H. With one h decaying to two photons and the other decaying to b quarks, the resonant signal may be discoverable above the hh continuum background for M(H)&lt;1  TeV. The product of the scalar and top Yukawa couplings can be measured to better than 10%-20% accuracy, and its sign can be inferred from the hh line shape via interference effects.

pubmed23n0842_15801

Digital imaging of colon tissue: method for evaluation of inflammation severity by spatial frequency features of the histological images.

The efficacy of histological analysis of colon sections used for evaluation of inflammation severity can be improved by means of digital imaging giving quantitative estimates of main diagnostic features. The aim of this study was to reveal most valuable diagnostic features reflecting inflammation severity in colon and elaborate the evaluation method for computer-aided diagnostics. Tissue specimens from 24 BALB/c mice and 15 patients were included in the study. Chronic and acute colon inflammation in mice was induced by oral administration of dextran sulphate sodium (DSS) solution, while mice in the control group did not get DSS. Human samples of inflamed colon tissue were obtained from patients with ulcerative colitis (n = 6). Non-inflamed colon tissue of control subjects (n = 9) was obtained from patients with irritable bowel syndrome or functional obstipation. Analysis of morphological changes in mice and human colon mucosa was performed using 4-μm haematoxylin-eosin (HE) sections. The features reflecting morphological changes in the images of colon mucosa were calculated by convolution of Gabor filter bank and array of pixel values. All features were generalized by calculating mean, histogram skewness and entropy of every image response. Principal component analysis was used to construct optimal representation of morphological changes. First principal component (PC1) was representing the major part of features variation (97 % in mice and 71 % in human specimens) and was selected as a measure of inflammation severity. Validation of new measure was performed by means of custom-made software realizing double blind comparison of differences in PC1 with expert's opinion about inflammation severity presented in two compared pictures. Overall accuracy of 80 % for mice and 67 % for human was reached. Principal component analysis of spatial frequency features of histological images may provide continuous scale estimation of inflammation severity of colon tissue.

Digital imaging of colon tissue: method for evaluation of inflammation severity by spatial frequency features of the histological images. The efficacy of histological analysis of colon sections used for evaluation of inflammation severity can be improved by means of digital imaging giving quantitative estimates of main diagnostic features. The aim of this study was to reveal most valuable diagnostic features reflecting inflammation severity in colon and elaborate the evaluation method for computer-aided diagnostics. Tissue specimens from 24 BALB/c mice and 15 patients were included in the study. Chronic and acute colon inflammation in mice was induced by oral administration of dextran sulphate sodium (DSS) solution, while mice in the control group did not get DSS. Human samples of inflamed colon tissue were obtained from patients with ulcerative colitis (n = 6). Non-inflamed colon tissue of control subjects (n = 9) was obtained from patients with irritable bowel syndrome or functional obstipation. Analysis of morphological changes in mice and human colon mucosa was performed using 4-μm haematoxylin-eosin (HE) sections. The features reflecting morphological changes in the images of colon mucosa were calculated by convolution of Gabor filter bank and array of pixel values. All features were generalized by calculating mean, histogram skewness and entropy of every image response. Principal component analysis was used to construct optimal representation of morphological changes. First principal component (PC1) was representing the major part of features variation (97 % in mice and 71 % in human specimens) and was selected as a measure of inflammation severity. Validation of new measure was performed by means of custom-made software realizing double blind comparison of differences in PC1 with expert's opinion about inflammation severity presented in two compared pictures. Overall accuracy of 80 % for mice and 67 % for human was reached. Principal component analysis of spatial frequency features of histological images may provide continuous scale estimation of inflammation severity of colon tissue.

pubmed23n1078_2033

Transcription factor EB promotes rheumatoid arthritis of Sprague-Dawley rats via regulating autophagy.

This study investigated the effect of autophagy-related gene transcription factor EB (TFEB) on the rheumatoid arthritis (RA) and explored whether TFEB regulated RA by autophagy. The Sprague-Dawley rats were divided into two groups (<in</i = 6). The rats were stimulated with the mixture of the type II collagen and Freund's adjuvant or PBS at the root of the tail. Results showed that swollen and deformed joints were discovered, the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were elevated, and hematoxylin and eosin staining showed the inflammatory cells infiltrate the synovial tissue in the RA rats, compared to the control group. Immunohistochemistry displayed that the expressions of TFEB and LC3B increased in the synovial tissues of RA rats, whereas p62 decreased. The silence of TFEB in the RA-fibroblast-like synoviocytes (RA-FLS) decreased the protein expressions of LC3B, compared to the siRNA NC group. Meanwhile, the activity of FLS was raised, whereas the levels of TNF-α and IL-6 decreased in RA-FLS with TFEB knockdown. In conclusion, our study revealed that TFEB plays a crucial role in the progress of RA by regulating autophagy, which might provide novel targets for the therapy of RA.

Transcription factor EB promotes rheumatoid arthritis of Sprague-Dawley rats via regulating autophagy. This study investigated the effect of autophagy-related gene transcription factor EB (TFEB) on the rheumatoid arthritis (RA) and explored whether TFEB regulated RA by autophagy. The Sprague-Dawley rats were divided into two groups (<in</i = 6). The rats were stimulated with the mixture of the type II collagen and Freund's adjuvant or PBS at the root of the tail. Results showed that swollen and deformed joints were discovered, the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were elevated, and hematoxylin and eosin staining showed the inflammatory cells infiltrate the synovial tissue in the RA rats, compared to the control group. Immunohistochemistry displayed that the expressions of TFEB and LC3B increased in the synovial tissues of RA rats, whereas p62 decreased. The silence of TFEB in the RA-fibroblast-like synoviocytes (RA-FLS) decreased the protein expressions of LC3B, compared to the siRNA NC group. Meanwhile, the activity of FLS was raised, whereas the levels of TNF-α and IL-6 decreased in RA-FLS with TFEB knockdown. In conclusion, our study revealed that TFEB plays a crucial role in the progress of RA by regulating autophagy, which might provide novel targets for the therapy of RA.

pubmed23n0864_13397

The Tactile Dimensions of Abstract Paintings: A Cross-Modal Study.

In our research, we tested for the existence of cross-modal visual and tactile associations in the experience of abstract art. Specifically, we measured the association of 60 abstract paintings with four couples of antonyms related to texture, such as warm or cold, smooth or rough, lightweight or heavy, soft or hard, investigating if the different modality of presentation on a computer screen (color versions: natural colors, inverted colors, black and white) gave rise to different associations relative to the four couples of opponent qualities. Second, we tested whether there might be differences between the ratings of the paintings when they were presented as images on a computer screen versus in real life at the museum. The results confirmed that associations between visual and tactile experience with such complex stimuli exist. In the case of the couple warm or cold, a significant inversion of associated qualities occurs when the images are presented in inverted colors as opposed to natural colors; furthermore, when presented in black and white, warm evaluations are "cooled down," but cold evaluations remain the same. The degree of smoothness could be considered not associated with the color versions. When seen in black and white, both the mean softness and the mean lightweight-ness of the paintings were reduced; however, in the last case, this effect was more evident for the most lightweight pictures. There is only a slight difference between the two presentations of the paintings as images presented on a computer screen and seen in real life, relative to the warm or cold and soft or hard dimensions. Of the four opponent qualities, the three pairs warm or cold, lightweight or heavy, and soft or hard showed the most interesting results in relation to the cross-modal associations.

The Tactile Dimensions of Abstract Paintings: A Cross-Modal Study. In our research, we tested for the existence of cross-modal visual and tactile associations in the experience of abstract art. Specifically, we measured the association of 60 abstract paintings with four couples of antonyms related to texture, such as warm or cold, smooth or rough, lightweight or heavy, soft or hard, investigating if the different modality of presentation on a computer screen (color versions: natural colors, inverted colors, black and white) gave rise to different associations relative to the four couples of opponent qualities. Second, we tested whether there might be differences between the ratings of the paintings when they were presented as images on a computer screen versus in real life at the museum. The results confirmed that associations between visual and tactile experience with such complex stimuli exist. In the case of the couple warm or cold, a significant inversion of associated qualities occurs when the images are presented in inverted colors as opposed to natural colors; furthermore, when presented in black and white, warm evaluations are "cooled down," but cold evaluations remain the same. The degree of smoothness could be considered not associated with the color versions. When seen in black and white, both the mean softness and the mean lightweight-ness of the paintings were reduced; however, in the last case, this effect was more evident for the most lightweight pictures. There is only a slight difference between the two presentations of the paintings as images presented on a computer screen and seen in real life, relative to the warm or cold and soft or hard dimensions. Of the four opponent qualities, the three pairs warm or cold, lightweight or heavy, and soft or hard showed the most interesting results in relation to the cross-modal associations.

pubmed23n1073_15075

Developing Personalized Models of Blood Pressure Estimation from Wearable Sensors Data Using Minimally-trained Domain Adversarial Neural Networks.

Blood pressure monitoring is an essential component of hypertension management and in the prediction of associated comorbidities. Blood pressure is a dynamic vital sign with frequent changes throughout a given day. Capturing blood pressure remotely and frequently (also known as ambulatory blood pressure monitoring) has traditionally been achieved by measuring blood pressure at discrete intervals using an inflatable cuff. However, there is growing interest in developing a cuffless ambulatory blood pressure monitoring system to measure blood pressure continuously. One such approach is by utilizing bioimpedance sensors to build regression models. A practical problem with this approach is that the amount of data required to confidently train such a regression model can be prohibitive. In this paper, we propose the application of the domain-adversarial training neural network (DANN) method on our multitask learning (MTL) blood pressure estimation model, allowing for knowledge transfer between subjects. Our proposed model obtains average root mean square error (RMSE) of 4.80 ± 0.74 mmHg for diastolic blood pressure and 7.34 ± 1.88 mmHg for systolic blood pressure when using three minutes of training data, 4.64 ± 0.60 mmHg and 7.10 ± 1.79 respectively when using four minutes of training data, and 4.48±0.57 mmHg and 6.79±1.70 respectively when using five minutes of training data. DANN improves training with minimal data in comparison to both directly training and to training with a pretrained model from another subject, decreasing RMSE by 0.19 to 0.26 mmHg (diastolic) and by 0.46 to 0.67 mmHg (systolic) in comparison to the best baseline models. We observe that four minutes of training data is the minimum requirement for our framework to exceed ISO standards within this cohort of patients.

Developing Personalized Models of Blood Pressure Estimation from Wearable Sensors Data Using Minimally-trained Domain Adversarial Neural Networks. Blood pressure monitoring is an essential component of hypertension management and in the prediction of associated comorbidities. Blood pressure is a dynamic vital sign with frequent changes throughout a given day. Capturing blood pressure remotely and frequently (also known as ambulatory blood pressure monitoring) has traditionally been achieved by measuring blood pressure at discrete intervals using an inflatable cuff. However, there is growing interest in developing a cuffless ambulatory blood pressure monitoring system to measure blood pressure continuously. One such approach is by utilizing bioimpedance sensors to build regression models. A practical problem with this approach is that the amount of data required to confidently train such a regression model can be prohibitive. In this paper, we propose the application of the domain-adversarial training neural network (DANN) method on our multitask learning (MTL) blood pressure estimation model, allowing for knowledge transfer between subjects. Our proposed model obtains average root mean square error (RMSE) of 4.80 ± 0.74 mmHg for diastolic blood pressure and 7.34 ± 1.88 mmHg for systolic blood pressure when using three minutes of training data, 4.64 ± 0.60 mmHg and 7.10 ± 1.79 respectively when using four minutes of training data, and 4.48±0.57 mmHg and 6.79±1.70 respectively when using five minutes of training data. DANN improves training with minimal data in comparison to both directly training and to training with a pretrained model from another subject, decreasing RMSE by 0.19 to 0.26 mmHg (diastolic) and by 0.46 to 0.67 mmHg (systolic) in comparison to the best baseline models. We observe that four minutes of training data is the minimum requirement for our framework to exceed ISO standards within this cohort of patients.

pubmed23n0919_19965

Biomarkers Associated with Atrial Fibrosis and Remodeling.

Atrial fibrillation is the most common rhythm disturbance encountered in clinical practice. Although often considered as solely arrhythmic in nature, current evidence has established that atrial myopathy constitutes both the substrate and the outcome of atrial fibrillation, thus initiating a vicious, self-perpetuating cycle. This myopathy is triggered by stress-induced (including pressure/volume overload, inflammation, oxidative stress) responses of atrial tissue, which in the long term become maladaptive, and combine elements of both structural, especially fibrosis, and electrical remodeling, with contemporary approaches yielding potentially useful biomarkers of these processes. Biomarker value becomes greater given the fact that they can both predict atrial fibrillation occurrence and treatment outcome. This mini-review will focus on the biomarkers of atrial remodeling (both electrical and structural) and fibrosis that have been validated in human studies, including biochemical, histological and imaging approaches.

Biomarkers Associated with Atrial Fibrosis and Remodeling. Atrial fibrillation is the most common rhythm disturbance encountered in clinical practice. Although often considered as solely arrhythmic in nature, current evidence has established that atrial myopathy constitutes both the substrate and the outcome of atrial fibrillation, thus initiating a vicious, self-perpetuating cycle. This myopathy is triggered by stress-induced (including pressure/volume overload, inflammation, oxidative stress) responses of atrial tissue, which in the long term become maladaptive, and combine elements of both structural, especially fibrosis, and electrical remodeling, with contemporary approaches yielding potentially useful biomarkers of these processes. Biomarker value becomes greater given the fact that they can both predict atrial fibrillation occurrence and treatment outcome. This mini-review will focus on the biomarkers of atrial remodeling (both electrical and structural) and fibrosis that have been validated in human studies, including biochemical, histological and imaging approaches.

pubmed23n0614_19533

HIV infection and AIDS 2: transmission and diagnosis.

This is a two-part unit on HIV infection and Aids. Part 1 reviewed the evolving global and national epidemiology of HIV infection and Aids. This second part examines routes of transmission, diagnosis and antiretroviral treatment.

HIV infection and AIDS 2: transmission and diagnosis. This is a two-part unit on HIV infection and Aids. Part 1 reviewed the evolving global and national epidemiology of HIV infection and Aids. This second part examines routes of transmission, diagnosis and antiretroviral treatment.

pubmed23n0522_13223

Human cytomegalovirus uracil DNA glycosylase associates with ppUL44 and accelerates the accumulation of viral DNA.

Human cytomegalovirus UL114 encodes a uracil-DNA glycosylase homolog that is highly conserved in all characterized herpesviruses that infect mammals. Previous studies demonstrated that the deletion of this nonessential gene delays significantly the onset of viral DNA synthesis and results in a prolonged replication cycle. The gene product, pUL114, also appears to be important in late phase DNA synthesis presumably by introducing single stranded breaks. A series of experiments was performed to formally assign the observed phenotype to pUL114 and to characterize the function of the protein in viral replication. A cell line expressing pUL114 complemented the observed phenotype of a UL114 deletion virus in trans, confirming that the observed defects were the result of a deficiency in this gene product. Stocks of recombinant viruses without elevated levels of uracil were produced in the complementing cells; however they retained the phenotype of poor growth in normal fibroblasts suggesting that poor replication was unrelated to uracil content of input genomes. Recombinant viruses expressing epitope tagged versions of this gene demonstrated that pUL114 was expressed at early times and that it localized to viral replication compartments. This protein also coprecipitated with the DNA polymerase processivity factor, ppUL44 suggesting that these proteins associate in infected cells. This apparent interaction did not appear to require other viral proteins since ppUL44 could recruit pUL114 to the nucleus in uninfected cells. An analysis of DNA replication kinetics revealed that the initial rate of DNA synthesis and the accumulation of progeny viral genomes were significantly reduced compared to the parent virus. These data suggest that pUL114 associates with ppUL44 and that it functions as part of the viral DNA replication complex to increase the efficiency of both early and late phase viral DNA synthesis.

Human cytomegalovirus uracil DNA glycosylase associates with ppUL44 and accelerates the accumulation of viral DNA. Human cytomegalovirus UL114 encodes a uracil-DNA glycosylase homolog that is highly conserved in all characterized herpesviruses that infect mammals. Previous studies demonstrated that the deletion of this nonessential gene delays significantly the onset of viral DNA synthesis and results in a prolonged replication cycle. The gene product, pUL114, also appears to be important in late phase DNA synthesis presumably by introducing single stranded breaks. A series of experiments was performed to formally assign the observed phenotype to pUL114 and to characterize the function of the protein in viral replication. A cell line expressing pUL114 complemented the observed phenotype of a UL114 deletion virus in trans, confirming that the observed defects were the result of a deficiency in this gene product. Stocks of recombinant viruses without elevated levels of uracil were produced in the complementing cells; however they retained the phenotype of poor growth in normal fibroblasts suggesting that poor replication was unrelated to uracil content of input genomes. Recombinant viruses expressing epitope tagged versions of this gene demonstrated that pUL114 was expressed at early times and that it localized to viral replication compartments. This protein also coprecipitated with the DNA polymerase processivity factor, ppUL44 suggesting that these proteins associate in infected cells. This apparent interaction did not appear to require other viral proteins since ppUL44 could recruit pUL114 to the nucleus in uninfected cells. An analysis of DNA replication kinetics revealed that the initial rate of DNA synthesis and the accumulation of progeny viral genomes were significantly reduced compared to the parent virus. These data suggest that pUL114 associates with ppUL44 and that it functions as part of the viral DNA replication complex to increase the efficiency of both early and late phase viral DNA synthesis.

pubmed23n0778_20891

Effects of regular exercise and nutritional guidance on body composition, blood pressure, muscle strength and health-related quality of life in community-dwelling Japanese women.

This study aimed to investigate the effects of 6-month regular exercise and nutritional guidance for body composition, blood pressure, muscle strength and health-related quality of life (HRQOL) in community-dwelling Japanese women aged 40-74 years. Participants were divided into an intervention group (n = 48) comprising women registered for health guidance and a control group without intervention (n = 66). The intervention group received 6-month exercise and nutritional guidance to modify lifestyle. Before and after the intervention period, body mass index (BMI), waist circumference, body fat percentage, blood pressure, muscle strength and HRQOL using the 36-item Short-Form Health Survey version 2 (SF-36) questionnaire were measured. At baseline, no significant differences were found between intervention and control groups. Waist circumference decreased significantly in the intervention group (from 82.4 to 79.9 cm) compared to the control group (from 80.5 to 79.7 cm). BMI and body fat percentage also decreased significantly more in the intervention group than in the control group. General health perception, vitality and social functioning in the SF-36 showed significantly greater improvements in the intervention group than in the control group. Six-month regular exercise and nutritional guidance had beneficial effects on body composition and health-related quality of life especially for mental components of SF-36. Based on these findings, our intervention was expected to provide benefits to mental components of HRQOL and facilitate sustained participation and motivation in modify lifestyles. As a result, beneficial effects on body composition might also be sustained.

Effects of regular exercise and nutritional guidance on body composition, blood pressure, muscle strength and health-related quality of life in community-dwelling Japanese women. This study aimed to investigate the effects of 6-month regular exercise and nutritional guidance for body composition, blood pressure, muscle strength and health-related quality of life (HRQOL) in community-dwelling Japanese women aged 40-74 years. Participants were divided into an intervention group (n = 48) comprising women registered for health guidance and a control group without intervention (n = 66). The intervention group received 6-month exercise and nutritional guidance to modify lifestyle. Before and after the intervention period, body mass index (BMI), waist circumference, body fat percentage, blood pressure, muscle strength and HRQOL using the 36-item Short-Form Health Survey version 2 (SF-36) questionnaire were measured. At baseline, no significant differences were found between intervention and control groups. Waist circumference decreased significantly in the intervention group (from 82.4 to 79.9 cm) compared to the control group (from 80.5 to 79.7 cm). BMI and body fat percentage also decreased significantly more in the intervention group than in the control group. General health perception, vitality and social functioning in the SF-36 showed significantly greater improvements in the intervention group than in the control group. Six-month regular exercise and nutritional guidance had beneficial effects on body composition and health-related quality of life especially for mental components of SF-36. Based on these findings, our intervention was expected to provide benefits to mental components of HRQOL and facilitate sustained participation and motivation in modify lifestyles. As a result, beneficial effects on body composition might also be sustained.

pubmed23n0837_7473

Operating a two-stage microbial fuel cell-constructed wetland for fuller wastewater treatment and more efficient electricity generation.

By integrating microbial fuel cells (MFCs) into constructed wetlands (CWs) the need and cost of building a reactor are eliminated, while CWs provide the simultaneous redox conditions required for optimum MFC performance. Two single-stage MFC-CWs, with dewatered alum sludge cake as the main wetland medium for enhanced phosphorus removal, were operated to determine the effects of electrode separation and flow regimes on power production and wastewater treatment. When the anode is buried and the cathode is at the air-water interface the system is inhibited by a large ohmic resistance resulting from the increased electrode separation. By placing the cathode directly above the anode and operating the system with simultaneous up-flow into the anode and down-flow into the cathode the ohmic resistance is reduced. The chemical oxygen demand (COD) removal efficiency was, however, reduced to 64% (compared with 79%). A two-stage system was subsequently run for fuller wastewater treatment and increased power production. The results indicate that a two-stage MFC-CW can increase the normalized energy recovery and improve removal efficiencies of COD, total nitrogen, NH4⁺, total phosphorus and reactive phosphorus to 93 ± 1.7%, 85 ± 5.2%, 90 ± 5.4%, 98 ± 5.3% and 99 ± 2.9%, respectively.

Operating a two-stage microbial fuel cell-constructed wetland for fuller wastewater treatment and more efficient electricity generation. By integrating microbial fuel cells (MFCs) into constructed wetlands (CWs) the need and cost of building a reactor are eliminated, while CWs provide the simultaneous redox conditions required for optimum MFC performance. Two single-stage MFC-CWs, with dewatered alum sludge cake as the main wetland medium for enhanced phosphorus removal, were operated to determine the effects of electrode separation and flow regimes on power production and wastewater treatment. When the anode is buried and the cathode is at the air-water interface the system is inhibited by a large ohmic resistance resulting from the increased electrode separation. By placing the cathode directly above the anode and operating the system with simultaneous up-flow into the anode and down-flow into the cathode the ohmic resistance is reduced. The chemical oxygen demand (COD) removal efficiency was, however, reduced to 64% (compared with 79%). A two-stage system was subsequently run for fuller wastewater treatment and increased power production. The results indicate that a two-stage MFC-CW can increase the normalized energy recovery and improve removal efficiencies of COD, total nitrogen, NH4⁺, total phosphorus and reactive phosphorus to 93 ± 1.7%, 85 ± 5.2%, 90 ± 5.4%, 98 ± 5.3% and 99 ± 2.9%, respectively.

pubmed23n0572_11104

Cyclin-dependent kinase 2 dependent phosphorylation of ATRIP regulates the G2-M checkpoint response to DNA damage.

The ATR-ATRIP kinase complex regulates cellular responses to DNA damage and replication stress. Mass spectrometry was used to identify phosphorylation sites on ATR and ATRIP to understand how the kinase complex is regulated by post-translational modifications. Two novel phosphorylation sites on ATRIP were identified, S224 and S239. Phosphopeptide-specific antibodies to S224 indicate that it is phosphorylated in a cell cycle-dependent manner. S224 matches a consensus site for cyclin-dependent kinase (CDK) phosphorylation and is phosphorylated by CDK2-cyclin A in vitro. S224 phosphorylation in cells is sensitive to CDK2 inhibitors. Mutation of S224 to alanine causes a defect in the ATR-ATRIP-dependent maintenance of the G(2)-M checkpoint to ionizing and UV radiation. Thus, ATRIP is a CDK2 substrate, and CDK2-dependent phosphorylation of S224 regulates the ability of ATR-ATRIP to promote cell cycle arrest in response to DNA damage.

Cyclin-dependent kinase 2 dependent phosphorylation of ATRIP regulates the G2-M checkpoint response to DNA damage. The ATR-ATRIP kinase complex regulates cellular responses to DNA damage and replication stress. Mass spectrometry was used to identify phosphorylation sites on ATR and ATRIP to understand how the kinase complex is regulated by post-translational modifications. Two novel phosphorylation sites on ATRIP were identified, S224 and S239. Phosphopeptide-specific antibodies to S224 indicate that it is phosphorylated in a cell cycle-dependent manner. S224 matches a consensus site for cyclin-dependent kinase (CDK) phosphorylation and is phosphorylated by CDK2-cyclin A in vitro. S224 phosphorylation in cells is sensitive to CDK2 inhibitors. Mutation of S224 to alanine causes a defect in the ATR-ATRIP-dependent maintenance of the G(2)-M checkpoint to ionizing and UV radiation. Thus, ATRIP is a CDK2 substrate, and CDK2-dependent phosphorylation of S224 regulates the ability of ATR-ATRIP to promote cell cycle arrest in response to DNA damage.

pubmed23n0715_6712

Experience of dental caries and its effects on early dental occlusion: a descriptive study.

Describe the occurrence of dental caries in a sample of pre-school children and school children, aged 3 and 12, and study the possible association between caries and malocclusion. We selected and analyzed the medical records of a sample of 588 patients who had their first dental examination at the Pediatric Dentistry Unit, Department of Oral and Maxillofacial Sciences of Policlinico Umberto I, "Sapienza" University of Rome. In the sample, 55.4% of the children had no decayed deciduous elements, while 44.6% had at least one decayed deciduous element. The prevalence of decayed permanent teeth was 10.2%, while 89.8% had no decayed permanent teeth. In the sample, 9.4% of the children showed advanced carious lesions, that needed tooth extraction and 6.6% needed a space maintainer for post-extractive interceptive treatment. In the sample, 26.7% of the examined patients had increased overjet, while 3.7% had decreased overjet and 25.4% of the sample had an increased overbite, 11, 2% had reduced over-bite values. A percentage over 10% of the sample had an anterior openbite in centric occlusion. The prevalence of posterior crossbite among entire samples was 19.8%. Although the incidence of caries disease was high in the selected samples, the study did not show a statistically significant association between caries and clinical orthodontic abnormalities, except for the association between the midline deviation and the severe carious diseases, necessitating extraction.

Experience of dental caries and its effects on early dental occlusion: a descriptive study. Describe the occurrence of dental caries in a sample of pre-school children and school children, aged 3 and 12, and study the possible association between caries and malocclusion. We selected and analyzed the medical records of a sample of 588 patients who had their first dental examination at the Pediatric Dentistry Unit, Department of Oral and Maxillofacial Sciences of Policlinico Umberto I, "Sapienza" University of Rome. In the sample, 55.4% of the children had no decayed deciduous elements, while 44.6% had at least one decayed deciduous element. The prevalence of decayed permanent teeth was 10.2%, while 89.8% had no decayed permanent teeth. In the sample, 9.4% of the children showed advanced carious lesions, that needed tooth extraction and 6.6% needed a space maintainer for post-extractive interceptive treatment. In the sample, 26.7% of the examined patients had increased overjet, while 3.7% had decreased overjet and 25.4% of the sample had an increased overbite, 11, 2% had reduced over-bite values. A percentage over 10% of the sample had an anterior openbite in centric occlusion. The prevalence of posterior crossbite among entire samples was 19.8%. Although the incidence of caries disease was high in the selected samples, the study did not show a statistically significant association between caries and clinical orthodontic abnormalities, except for the association between the midline deviation and the severe carious diseases, necessitating extraction.

pubmed23n0715_12813

Trigeminal neuralgia and persistent trigeminal artery.

We report a case of trigeminal neuralgia caused by persistent trigeminal artery (PTA) associated with asymptomatic left temporal cavernoma. Our patient presented unstable blood hypertension and the pain of typical trigeminal neuralgia over the second and third divisions of the nerve in the right side of the face. The attacks were often precipitated during physical exertion. MRI and Angio-MRI revealed the persistent carotid basilar anastomosis and occasionally left parietal cavernoma. After drug treatment of blood hypertension, spontaneous recovery of neuralgia was observed and we planned surgical treatment of left temporal cavernoma.

Trigeminal neuralgia and persistent trigeminal artery. We report a case of trigeminal neuralgia caused by persistent trigeminal artery (PTA) associated with asymptomatic left temporal cavernoma. Our patient presented unstable blood hypertension and the pain of typical trigeminal neuralgia over the second and third divisions of the nerve in the right side of the face. The attacks were often precipitated during physical exertion. MRI and Angio-MRI revealed the persistent carotid basilar anastomosis and occasionally left parietal cavernoma. After drug treatment of blood hypertension, spontaneous recovery of neuralgia was observed and we planned surgical treatment of left temporal cavernoma.

pubmed23n0694_2687

The effect of prophylactic lamivudine on hepatitis B virus reactivation in HBsAg-positive patients with diffuse large B-cell lymphoma undergoing prolonged rituximab therapy.

The association of prolonged rituximab therapy and hepatitis B virus (HBV) reactivation in diffuse large B-cell lymphoma (DLBCL) and the role of lamivudine prophylaxis remain undefined. The prevalence and mortality of HBV reactivation in HBsAg-positive patients with DLBCL undergoing rituximab-based treatment, who received prophylactic treatment with or without lamivudine, were retrospectively analyzed. From January 2003 to December 2009, there were 50 patients enrolled in the study, among of which 30 received the prophylactic treatment of lamivudine and 20 without prophylactic treatment of lamivudine. Among of the 50 patients, seven patients received further rituximab maintenance, once every 3 months for 2 years. Compared with lamivudine treatment group, it showed that there was significantly higher prevalence of HBV reactivation (60.0% vs 13.3%, P = .001), severe hepatitis (45.0% vs 6.7%, P = .004), and mortality (25.0% vs 3.3%, P = .032) in non-lamivudine prophylactic group; however, there was no statistically significant difference in the HBV DNA levels at reactivation (3.94 × 10(6) vs 8.30 × 10(5) copies/ml, P = .47) and the time from first dose of rituximab to HBV reactivation(207 vs 386 days, P = .28). For patients undergoing further rituximab maintanence treatment, the prevalence and mortality of HBV reactivation were 71.4 and 28.6%, respectively. The prevalence and mortality of HBV reactivation are 66.7% vs 75.0% (P = 1.00) and 0 vs 50.0% (P = .43) in lamivudine prophylactic and non-lamivudine prophylactic groups, respectively. The effect of lamivudine prophylaxis on preventing HBV reactivation was found to be less in patients undergoing longer duration of rituximab treatment. A longer duration of rituximab treatment contributed to higher morbidity and mortality of HBV reactivation in HbsAg-positive patients with DLBCL. Further study is warranted for the optimal management of hepatitis caused by HBV reactivation.

The effect of prophylactic lamivudine on hepatitis B virus reactivation in HBsAg-positive patients with diffuse large B-cell lymphoma undergoing prolonged rituximab therapy. The association of prolonged rituximab therapy and hepatitis B virus (HBV) reactivation in diffuse large B-cell lymphoma (DLBCL) and the role of lamivudine prophylaxis remain undefined. The prevalence and mortality of HBV reactivation in HBsAg-positive patients with DLBCL undergoing rituximab-based treatment, who received prophylactic treatment with or without lamivudine, were retrospectively analyzed. From January 2003 to December 2009, there were 50 patients enrolled in the study, among of which 30 received the prophylactic treatment of lamivudine and 20 without prophylactic treatment of lamivudine. Among of the 50 patients, seven patients received further rituximab maintenance, once every 3 months for 2 years. Compared with lamivudine treatment group, it showed that there was significantly higher prevalence of HBV reactivation (60.0% vs 13.3%, P = .001), severe hepatitis (45.0% vs 6.7%, P = .004), and mortality (25.0% vs 3.3%, P = .032) in non-lamivudine prophylactic group; however, there was no statistically significant difference in the HBV DNA levels at reactivation (3.94 × 10(6) vs 8.30 × 10(5) copies/ml, P = .47) and the time from first dose of rituximab to HBV reactivation(207 vs 386 days, P = .28). For patients undergoing further rituximab maintanence treatment, the prevalence and mortality of HBV reactivation were 71.4 and 28.6%, respectively. The prevalence and mortality of HBV reactivation are 66.7% vs 75.0% (P = 1.00) and 0 vs 50.0% (P = .43) in lamivudine prophylactic and non-lamivudine prophylactic groups, respectively. The effect of lamivudine prophylaxis on preventing HBV reactivation was found to be less in patients undergoing longer duration of rituximab treatment. A longer duration of rituximab treatment contributed to higher morbidity and mortality of HBV reactivation in HbsAg-positive patients with DLBCL. Further study is warranted for the optimal management of hepatitis caused by HBV reactivation.

pubmed23n0073_9782

Improving the adaptation of denture bases by anchorage to the casts: a comparative study.

The purpose of this study was to investigate the adaptation of complete maxillary dentures to stone casts processed by the conventional, and two anchoring, methods. The anchoring methods used holes drilled on the cast and a special flange extended onto the posterior aspect of the maxillary cast. Both anchoring methods improved the adaptation of denture bases by minimizing the discrepancy between the denture base and cast. The greatest discrepancies observed in all methods were at the central portion of the posterior border.

Improving the adaptation of denture bases by anchorage to the casts: a comparative study. The purpose of this study was to investigate the adaptation of complete maxillary dentures to stone casts processed by the conventional, and two anchoring, methods. The anchoring methods used holes drilled on the cast and a special flange extended onto the posterior aspect of the maxillary cast. Both anchoring methods improved the adaptation of denture bases by minimizing the discrepancy between the denture base and cast. The greatest discrepancies observed in all methods were at the central portion of the posterior border.

pubmed23n1080_23571

Formulating a Meaningful and Comprehensive Placental Phenotypic Classification.

While many placental lesions have been identified and defined, the significance of multiple overlapping lesions has not been addressed. The purpose of our analysis was to evaluate overlapping patterns of placental pathology and determine meaningful phenotypes associated with adverse birth outcomes. Placental pathology reports were obtained from a single hospital between 2009 and 2018. Placental lesions were grouped into four major categories: acute inflammation (AI), chronic inflammation (CI), maternal vascular malperfusion (MVM), and fetal vascular malperfusion (FVM). Within each category, lesions were classified as not present, low grade or high grade. Combinations of pathologies were evaluated in relation to preterm birth (&lt;37 weeks) and small for gestational age (SGA) infant (birthweight &lt;10th percentile). During the study period, 19,027 placentas were reviewed by pathologists. Results from interaction models indicate that MVM and MVM in combination with CI and/or FVM are associated with the greatest odds of SGA infant and PTB. When incorporating grade, we identified 21 phenotype groups, each with characteristic associations with the SGA infant and patterns of PTB. We have developed a comprehensive and meaningful placental phenotype that incorporates severity and multiplicity of placental lesions. We have also developed a web application to facilitate phenotype determination (https://placentaexpression.shinyapps.io/phenotype).

Formulating a Meaningful and Comprehensive Placental Phenotypic Classification. While many placental lesions have been identified and defined, the significance of multiple overlapping lesions has not been addressed. The purpose of our analysis was to evaluate overlapping patterns of placental pathology and determine meaningful phenotypes associated with adverse birth outcomes. Placental pathology reports were obtained from a single hospital between 2009 and 2018. Placental lesions were grouped into four major categories: acute inflammation (AI), chronic inflammation (CI), maternal vascular malperfusion (MVM), and fetal vascular malperfusion (FVM). Within each category, lesions were classified as not present, low grade or high grade. Combinations of pathologies were evaluated in relation to preterm birth (&lt;37 weeks) and small for gestational age (SGA) infant (birthweight &lt;10th percentile). During the study period, 19,027 placentas were reviewed by pathologists. Results from interaction models indicate that MVM and MVM in combination with CI and/or FVM are associated with the greatest odds of SGA infant and PTB. When incorporating grade, we identified 21 phenotype groups, each with characteristic associations with the SGA infant and patterns of PTB. We have developed a comprehensive and meaningful placental phenotype that incorporates severity and multiplicity of placental lesions. We have also developed a web application to facilitate phenotype determination (https://placentaexpression.shinyapps.io/phenotype).

pubmed23n0767_2374

[Training in emergency situations through medical simulation].

Alongside conventional teaching, simulation is an effective training technique. It comprises a series of techniques enabling experiences in real situations to be replaced by those in equivalent situations in an immersive and interactive way This type of training does not enable errors to be totally avoided, but helps to reduce the consequences.

[Training in emergency situations through medical simulation]. Alongside conventional teaching, simulation is an effective training technique. It comprises a series of techniques enabling experiences in real situations to be replaced by those in equivalent situations in an immersive and interactive way This type of training does not enable errors to be totally avoided, but helps to reduce the consequences.

pubmed23n0732_5159

Surface dilational moduli of poly (ethylene oxide), poly (methyl methacrylate), and their blend films.

Surface dilational moduli of poly (ethylene oxide) (PEO), poly (methyl methacrylate) (PMMA), and compatible PEO/PMMA blend films spread at the air-water interface were investigated as a function of surface concentration. The surface dilational modulus of an expanded PEO film increased as the surface concentration increased to 0.4 mg/m(2), which corresponds to the limiting surface area of PEO. After peaking at this value, the surface dilational modulus decreased with an increase in the PEO concentration. Lissajous orbits of PEO films exhibited positive hysteresis loops for all surface concentration ranges. On the other hand, the surface dilational modulus of a condensed PMMA film steeply increased as the surface concentration increased. Lissajous orbits of PMMA films changed from positive hysteresis loops to negative loops at the surface concentration at which the surface pressure reached in the plateau region. The magnitude of the surface dilational modulus of PMMA was larger than that of PEO at a fixed surface concentration. The surface dilational moduli of the PEO/PMMA blend films increased with the total surface concentration and their magnitudes were less than those of the individual PMMA films and larger than those of the individual PEO films at fixed surface concentrations. Lissajous orbits of the PEO/PMMA blend films also changed from positive hysteresis loops to negative loops beyond the surface concentration at which the plateau surface pressure of PEO was attained.

Surface dilational moduli of poly (ethylene oxide), poly (methyl methacrylate), and their blend films. Surface dilational moduli of poly (ethylene oxide) (PEO), poly (methyl methacrylate) (PMMA), and compatible PEO/PMMA blend films spread at the air-water interface were investigated as a function of surface concentration. The surface dilational modulus of an expanded PEO film increased as the surface concentration increased to 0.4 mg/m(2), which corresponds to the limiting surface area of PEO. After peaking at this value, the surface dilational modulus decreased with an increase in the PEO concentration. Lissajous orbits of PEO films exhibited positive hysteresis loops for all surface concentration ranges. On the other hand, the surface dilational modulus of a condensed PMMA film steeply increased as the surface concentration increased. Lissajous orbits of PMMA films changed from positive hysteresis loops to negative loops at the surface concentration at which the surface pressure reached in the plateau region. The magnitude of the surface dilational modulus of PMMA was larger than that of PEO at a fixed surface concentration. The surface dilational moduli of the PEO/PMMA blend films increased with the total surface concentration and their magnitudes were less than those of the individual PMMA films and larger than those of the individual PEO films at fixed surface concentrations. Lissajous orbits of the PEO/PMMA blend films also changed from positive hysteresis loops to negative loops beyond the surface concentration at which the plateau surface pressure of PEO was attained.

pubmed23n0952_22670

Rotational Thromboelastometry Rapidly Predicts Thrombocytopenia and Hypofibrinogenemia During Neonatal Cardiopulmonary Bypass.

Thrombocytopenia and hypofibrinogenemia during neonatal cardiopulmonary bypass (CPB) contribute to bleeding and morbidity. Rotational thromboelastometry (ROTEM) is a viscoelastic assay with a rapid turnaround time. Data validating ROTEM during neonatal cardiac surgery remain limited. This study examined perioperative hemostatic trends in neonates treated with standardized platelet and cryoprecipitate transfusion during CPB. We hypothesized that ROTEM would predict thrombocytopenia, hypofibrinogenemia, and the correction thereof. Forty-four neonates undergoing CPB were included in this prospective observational study. Blood samples were obtained at Baseline, On CPB, Post-CPB, and Postoperative. The ROTEM analysis included extrinsically activated (Extem) and fibrinogen-specific (Fibtem) assays. Platelet-specific thromboelastometry (Pltem) values were calculated. Platelet and cryoprecipitate transfusion was initiated prior to termination of CPB. Platelet count and Extem amplitude decreased significantly On CPB ( P &lt; .0001), increased significantly Post-CPB ( P &lt; .0001), and Postoperative values were not significantly different from Baseline. Extem amplitude at 10 minutes (A10) &gt; 46.5 mm (AUC = 0.941) and Pltem A10 &gt; 37.5 mm [area under curve (AUC) = 0.960] predicted platelet count &gt; 100 × 10<sup3</sup/μL, and they highly correlated with platelet count ( R = 0.89 and R = 0.90, respectively). Fibrinogen concentration and Fibtem amplitude decreased significantly On CPB ( P ≤ .0001) and normalized after cryoprecipitate transfusion. Fibtem A10 &gt; 9.5 mm predicted fibrinogen &gt;200 mg/dL (AUC = 0.817), but it correlated less well with fibrinogen concentration ( R = 0.65). ROTEM analysis during neonatal cardiac surgery is sensitive and specific for thrombocytopenia and hypofibrinogenemia, identifying deficits within 10 minutes. Platelet and cryoprecipitate transfusion during neonatal CPB normalizes platelet count, fibrinogen level, and ROTEM amplitudes.

Rotational Thromboelastometry Rapidly Predicts Thrombocytopenia and Hypofibrinogenemia During Neonatal Cardiopulmonary Bypass. Thrombocytopenia and hypofibrinogenemia during neonatal cardiopulmonary bypass (CPB) contribute to bleeding and morbidity. Rotational thromboelastometry (ROTEM) is a viscoelastic assay with a rapid turnaround time. Data validating ROTEM during neonatal cardiac surgery remain limited. This study examined perioperative hemostatic trends in neonates treated with standardized platelet and cryoprecipitate transfusion during CPB. We hypothesized that ROTEM would predict thrombocytopenia, hypofibrinogenemia, and the correction thereof. Forty-four neonates undergoing CPB were included in this prospective observational study. Blood samples were obtained at Baseline, On CPB, Post-CPB, and Postoperative. The ROTEM analysis included extrinsically activated (Extem) and fibrinogen-specific (Fibtem) assays. Platelet-specific thromboelastometry (Pltem) values were calculated. Platelet and cryoprecipitate transfusion was initiated prior to termination of CPB. Platelet count and Extem amplitude decreased significantly On CPB ( P &lt; .0001), increased significantly Post-CPB ( P &lt; .0001), and Postoperative values were not significantly different from Baseline. Extem amplitude at 10 minutes (A10) &gt; 46.5 mm (AUC = 0.941) and Pltem A10 &gt; 37.5 mm [area under curve (AUC) = 0.960] predicted platelet count &gt; 100 × 10<sup3</sup/μL, and they highly correlated with platelet count ( R = 0.89 and R = 0.90, respectively). Fibrinogen concentration and Fibtem amplitude decreased significantly On CPB ( P ≤ .0001) and normalized after cryoprecipitate transfusion. Fibtem A10 &gt; 9.5 mm predicted fibrinogen &gt;200 mg/dL (AUC = 0.817), but it correlated less well with fibrinogen concentration ( R = 0.65). ROTEM analysis during neonatal cardiac surgery is sensitive and specific for thrombocytopenia and hypofibrinogenemia, identifying deficits within 10 minutes. Platelet and cryoprecipitate transfusion during neonatal CPB normalizes platelet count, fibrinogen level, and ROTEM amplitudes.

pubmed23n0346_18556

Deletion of type IIalpha regulatory subunit delocalizes protein kinase A in mouse sperm without affecting motility or fertilization.

Cyclic AMP stimulates sperm motility in a variety of mammalian species, but the molecular details of the intracellular signaling pathway responsible for this effect are unclear. The type IIalpha isoform of protein kinase A (PKA) is induced late in spermatogenesis and is thought to localize PKA to the flagellar apparatus where it binds cAMP and stimulates motility. A targeted disruption of the type IIalpha regulatory subunit (RIIalpha) gene allowed us to examine the role of PKA localization in sperm motility and fertility. In wild type sperm, PKA is found primarily in the detergent-resistant particulate fraction and localizes to the mitochondrial-containing midpiece and the principal piece. In mutant sperm, there is a compensatory increase in RIalpha protein and a dramatic relocalization of PKA such that the majority of the holoenzyme now appears in the soluble fraction and colocalizes with the cytoplasmic droplet. Unexpectedly the RIIalpha mutant mice are fertile and have no significant changes in sperm motility. Our results demonstrate that the highly localized pattern of PKA seen in mature sperm is not essential for motility or fertilization.

Deletion of type IIalpha regulatory subunit delocalizes protein kinase A in mouse sperm without affecting motility or fertilization. Cyclic AMP stimulates sperm motility in a variety of mammalian species, but the molecular details of the intracellular signaling pathway responsible for this effect are unclear. The type IIalpha isoform of protein kinase A (PKA) is induced late in spermatogenesis and is thought to localize PKA to the flagellar apparatus where it binds cAMP and stimulates motility. A targeted disruption of the type IIalpha regulatory subunit (RIIalpha) gene allowed us to examine the role of PKA localization in sperm motility and fertility. In wild type sperm, PKA is found primarily in the detergent-resistant particulate fraction and localizes to the mitochondrial-containing midpiece and the principal piece. In mutant sperm, there is a compensatory increase in RIalpha protein and a dramatic relocalization of PKA such that the majority of the holoenzyme now appears in the soluble fraction and colocalizes with the cytoplasmic droplet. Unexpectedly the RIIalpha mutant mice are fertile and have no significant changes in sperm motility. Our results demonstrate that the highly localized pattern of PKA seen in mature sperm is not essential for motility or fertilization.

pubmed23n0097_8642

Rapid prenatal diagnosis of beta thalassemia using DNA amplification and nonradioactive probes.

We used in vitro DNA amplification by the polymerase chain reaction and nonradioactive probes for prenatal diagnosis of beta thalassemia in Chinese from the Guangdong province. Exact molecular diagnoses were made in all 20 fetuses studied over a 6-month period. We conclude that this method of prenatal diagnosis for beta thalassemia is a viable approach in many parts of the world where this disease is common.

Rapid prenatal diagnosis of beta thalassemia using DNA amplification and nonradioactive probes. We used in vitro DNA amplification by the polymerase chain reaction and nonradioactive probes for prenatal diagnosis of beta thalassemia in Chinese from the Guangdong province. Exact molecular diagnoses were made in all 20 fetuses studied over a 6-month period. We conclude that this method of prenatal diagnosis for beta thalassemia is a viable approach in many parts of the world where this disease is common.

pubmed23n1053_25455

Posterior approaches to the ankle - an analysis of 3 approaches for access to the posterior malleolar fracture.

An anatomical study to determine what degree of access to the posterior distal tibia could be gained by using 3 different approaches; the posterolateral, the posteromedial and the medial posteromedial approaches. A comparison study, between the anatomical dissection of 7 fresh frozen cadaveric lower legs and image analysis of CT data of posterior malleolar fractures from a prospectively collected database was conducted. All fractures have been classified using the Mason and Molloy classification. In comparing the posterior malleolar fracture fragment width to distal tibia width, the posterolateral fragment encompasses 60.1% (95% CI 56.8, 63.3) of the total width of the tibia. If the posteromedial fragment is included the fragments encompass the entire distal tibia (100%). In type 3 fractures, 81.4% (95% CI 75.5, 87.1) of the distal tibia width is involved. When comparing the fracture width to the approach, no approach achieves a complete exposure of the type 2B or 3 fracture patterns. The overall surface area of the type 2B and 3 fractures, is significantly greater than all the approaches. Considering the lateral to medial extent of the fracture, the posterolateral fragment mean width is 33% greater than what can be exposed by the posterolateral approach (mean 24.9 vs 16.8mm). In type 2B and 3 fractures, the horizontal exposure reduces to 39.8% and 47.6% respectively. In comparison, the PM approach exposes 47.6% of the type 2B fracture pattern and 57.1% of the type 3 fracture pattern and allows a preferable angle for hardware insertion. The MPM approach does not expose any of the posterolateral fragments in this study, however it does expose 92% (mean 21.9 vs. 23.8mm) of the medial to lateral width of a posteromedial fragment of a type 2B fracture. Each approach allows access to different parts and amounts of the posterior tibia. An understanding of and utilisation of these approaches can lead to adequate exposure for fixation of most posterior malleolus fracture patterns seen.

Posterior approaches to the ankle - an analysis of 3 approaches for access to the posterior malleolar fracture. An anatomical study to determine what degree of access to the posterior distal tibia could be gained by using 3 different approaches; the posterolateral, the posteromedial and the medial posteromedial approaches. A comparison study, between the anatomical dissection of 7 fresh frozen cadaveric lower legs and image analysis of CT data of posterior malleolar fractures from a prospectively collected database was conducted. All fractures have been classified using the Mason and Molloy classification. In comparing the posterior malleolar fracture fragment width to distal tibia width, the posterolateral fragment encompasses 60.1% (95% CI 56.8, 63.3) of the total width of the tibia. If the posteromedial fragment is included the fragments encompass the entire distal tibia (100%). In type 3 fractures, 81.4% (95% CI 75.5, 87.1) of the distal tibia width is involved. When comparing the fracture width to the approach, no approach achieves a complete exposure of the type 2B or 3 fracture patterns. The overall surface area of the type 2B and 3 fractures, is significantly greater than all the approaches. Considering the lateral to medial extent of the fracture, the posterolateral fragment mean width is 33% greater than what can be exposed by the posterolateral approach (mean 24.9 vs 16.8mm). In type 2B and 3 fractures, the horizontal exposure reduces to 39.8% and 47.6% respectively. In comparison, the PM approach exposes 47.6% of the type 2B fracture pattern and 57.1% of the type 3 fracture pattern and allows a preferable angle for hardware insertion. The MPM approach does not expose any of the posterolateral fragments in this study, however it does expose 92% (mean 21.9 vs. 23.8mm) of the medial to lateral width of a posteromedial fragment of a type 2B fracture. Each approach allows access to different parts and amounts of the posterior tibia. An understanding of and utilisation of these approaches can lead to adequate exposure for fixation of most posterior malleolus fracture patterns seen.

pubmed23n0574_3467

Metric analysis of the hard palate in children with Down syndrome: a comparative study.

The hard palate is viewed as playing an important role in the passive articulation of speech. Its probable role in the defective articulation of speech in individuals with Down syndrome has been examined in the present study. In individuals with Down syndrome, the hard palate is highly arched, constricted, and narrow and stair type with malformed misaligned teeth and a large and fissured tongue. As a result good palato-lingual contact is not achieved, with resulting defective articulation. Using orthodontic and prosthodontic principles could modify this situation, i.e. the anatomy of the hard palate. The altered palatal contour may give better placing to the tongue, leading to improved palato-lingual contact and articulation. The dimensional parameters measured were: average linear width (AVL), average curvilinear width (AVCL), average height (AVH) at different planes; average antero-posterior length (AAP), average volume (V), palatal arch length (PAL), and palatal index (PI). The findings were compared with those of controls of the same age and sex. The AVL, AVCL, AAP, PAL, V and PI values of patients with Down syndrome were found to be less than the corresponding values of controls and the average height values of patients with Down syndrome were greater than the corresponding values of controls. Statistical significance was observed in all measurements between the controls and the patients with Down syndrome, especially in those concerning the height and the volume of the oral cavity. Observations from this study have suggested that prostheses might be designed to modify the palatal anatomy and produce better articulation in people with Down syndrome.

Metric analysis of the hard palate in children with Down syndrome: a comparative study. The hard palate is viewed as playing an important role in the passive articulation of speech. Its probable role in the defective articulation of speech in individuals with Down syndrome has been examined in the present study. In individuals with Down syndrome, the hard palate is highly arched, constricted, and narrow and stair type with malformed misaligned teeth and a large and fissured tongue. As a result good palato-lingual contact is not achieved, with resulting defective articulation. Using orthodontic and prosthodontic principles could modify this situation, i.e. the anatomy of the hard palate. The altered palatal contour may give better placing to the tongue, leading to improved palato-lingual contact and articulation. The dimensional parameters measured were: average linear width (AVL), average curvilinear width (AVCL), average height (AVH) at different planes; average antero-posterior length (AAP), average volume (V), palatal arch length (PAL), and palatal index (PI). The findings were compared with those of controls of the same age and sex. The AVL, AVCL, AAP, PAL, V and PI values of patients with Down syndrome were found to be less than the corresponding values of controls and the average height values of patients with Down syndrome were greater than the corresponding values of controls. Statistical significance was observed in all measurements between the controls and the patients with Down syndrome, especially in those concerning the height and the volume of the oral cavity. Observations from this study have suggested that prostheses might be designed to modify the palatal anatomy and produce better articulation in people with Down syndrome.

pubmed23n0905_19192

GATA2 expression and biochemical recurrence following salvage radiation therapy for relapsing prostate cancer.

High GATA2 expression has been associated with an increased risk of poor clinical outcomes after radical prostatectomy; however, this has not been studied in relation to risk of biochemical recurrence (BCR) after salvage radiation therapy (SRT) for recurrent prostate cancer after radical prostatectomy. Our aim was to evaluate the association between protein expression levels of GATA2 in primary prostate cancer tumour samples and the risk of BCR after SRT. 109 males who were treated with SRT were included. The percentage of cells with nuclear staining and GATA2 staining intensity were both measured. These two measures were multiplied together to obtain a GATA2 H-score (range 0-12) which was our primary GATA2 staining measure. In unadjusted analysis, the risk of BCR was higher for patients with a GATA2 H-score &gt;4 (hazard ratio = 2.04, p = 0.033). In multivariable analysis adjusting for SRT dose, pre-SRT PSA, pathological tumour stage and Gleason score, this association weakened substantially (hazard ratio = 1.45, p = 0.31). This lack of an independent association with BCR appears to be the result of correlations between GATA2 H-score &gt;4 and higher pre-SRT PSA (p = 0.021), higher Gleason score (p = 0.044) and more severe pathological tumour stage (p = 0.068). Higher levels of GATA2 expression appear to be a marker of prostate cancer severity; however, these do not provide independent prognostic information regarding BCR beyond that of validated clinicopathological risk factors. Advances in knowledge: A higher GATA2 expression level appears to be correlated with known measures of prostate cancer severity and therefore is likely not an independent marker of outcome after SRT.

GATA2 expression and biochemical recurrence following salvage radiation therapy for relapsing prostate cancer. High GATA2 expression has been associated with an increased risk of poor clinical outcomes after radical prostatectomy; however, this has not been studied in relation to risk of biochemical recurrence (BCR) after salvage radiation therapy (SRT) for recurrent prostate cancer after radical prostatectomy. Our aim was to evaluate the association between protein expression levels of GATA2 in primary prostate cancer tumour samples and the risk of BCR after SRT. 109 males who were treated with SRT were included. The percentage of cells with nuclear staining and GATA2 staining intensity were both measured. These two measures were multiplied together to obtain a GATA2 H-score (range 0-12) which was our primary GATA2 staining measure. In unadjusted analysis, the risk of BCR was higher for patients with a GATA2 H-score &gt;4 (hazard ratio = 2.04, p = 0.033). In multivariable analysis adjusting for SRT dose, pre-SRT PSA, pathological tumour stage and Gleason score, this association weakened substantially (hazard ratio = 1.45, p = 0.31). This lack of an independent association with BCR appears to be the result of correlations between GATA2 H-score &gt;4 and higher pre-SRT PSA (p = 0.021), higher Gleason score (p = 0.044) and more severe pathological tumour stage (p = 0.068). Higher levels of GATA2 expression appear to be a marker of prostate cancer severity; however, these do not provide independent prognostic information regarding BCR beyond that of validated clinicopathological risk factors. Advances in knowledge: A higher GATA2 expression level appears to be correlated with known measures of prostate cancer severity and therefore is likely not an independent marker of outcome after SRT.

pubmed23n0110_13450

Effects of low X-ray doses in Saccharomyces cerevisiae.

Three strains of Saccharomyces cerevisiae with different capacities for repair of radiation damage (RAD, rad18, and rad52) have been tested for their colony forming ability (CFA) and growth rates after application of small X-ray doses from 3.8 mGy to 40 Gy. There was no reproducible increase in CFA observable after application of doses between 3.8 mGy and 4.7 Gy. X-ray doses of 40 Gy causing an inactivation of CFA from 90% to 50%, depending on the repair capacity of the strains used, caused a reduced increase in optical density during 2 h buffer treatment in comparison to unirradiated cells. This reduction however, is reversible as soon as the cells are transferred into nutrient medium. One hour after transfer into growth medium the portions of cells with large buds (G2 and M phase) and cells with small buds (S phase) are drastically different in irradiated cells from those obtained in unirradiated cells. The time necessary for separation of mother and daughter cells is prolonged by X-ray irradiation and the formation of new buds is retarded.

Effects of low X-ray doses in Saccharomyces cerevisiae. Three strains of Saccharomyces cerevisiae with different capacities for repair of radiation damage (RAD, rad18, and rad52) have been tested for their colony forming ability (CFA) and growth rates after application of small X-ray doses from 3.8 mGy to 40 Gy. There was no reproducible increase in CFA observable after application of doses between 3.8 mGy and 4.7 Gy. X-ray doses of 40 Gy causing an inactivation of CFA from 90% to 50%, depending on the repair capacity of the strains used, caused a reduced increase in optical density during 2 h buffer treatment in comparison to unirradiated cells. This reduction however, is reversible as soon as the cells are transferred into nutrient medium. One hour after transfer into growth medium the portions of cells with large buds (G2 and M phase) and cells with small buds (S phase) are drastically different in irradiated cells from those obtained in unirradiated cells. The time necessary for separation of mother and daughter cells is prolonged by X-ray irradiation and the formation of new buds is retarded.

pubmed23n0094_12869

[Effect of ascorbic acid on 25-hydroxyvitamin D3 metabolism in the kidneys and 1,25-dihydroxyvitamin D3 reception in the small intestine mucosa of guinea pigs].

Ascorbic acid deficiency in vitamin D-supplied guinea pigs caused a moderate decrease of Ca in the blood and osseous tissue, a 1.5-fold decrease of 2.5-hydroxyvitamin D (25-OH D) in blood serum, a 2-fold decrease of the 25-OH D 1-hydroxylase activity in kidneys and a 1.6-fold increase of the 24-hydroxylase activity. The concentration of 1.25-dihydroxyvitamin D3 (1.25-(OH)2D3) nuclear receptors in small intestinal mucosa diminished by 20-30%; in this case the percentage of occupied hormone receptors reduced from 11.8 to 8.6%. The affinity of receptors for 1.25-(OH)2D3 did not change thereby (Kd = 0.25-0.26 nM; Kd2 = 0.06-0.10 nM). At the same time the value of cooperativity coefficient showed a decrease-from 1.7 to 1.4, which was accompanied by a reduction of the maximum capacity of receptors (1.2-1.5-fold). Vitamin C depletion augmented the manifestation of vitamin D deficiency in guinea pigs and impeded their correction after administration of cholecalciferol. This markedly retarded the restoration of the 25-OH D level in the blood as well as the number of occupied and unoccupied nuclear receptors for 1.25-(OH)2D3. The experimental results illustrate the effects of ascorbic acid on the vitamin D hormonal system function, which is manifested both at the level of 1.25-(OH)2D3 synthesis in the kidneys and of its receptor binding in target tissues.

[Effect of ascorbic acid on 25-hydroxyvitamin D3 metabolism in the kidneys and 1,25-dihydroxyvitamin D3 reception in the small intestine mucosa of guinea pigs]. Ascorbic acid deficiency in vitamin D-supplied guinea pigs caused a moderate decrease of Ca in the blood and osseous tissue, a 1.5-fold decrease of 2.5-hydroxyvitamin D (25-OH D) in blood serum, a 2-fold decrease of the 25-OH D 1-hydroxylase activity in kidneys and a 1.6-fold increase of the 24-hydroxylase activity. The concentration of 1.25-dihydroxyvitamin D3 (1.25-(OH)2D3) nuclear receptors in small intestinal mucosa diminished by 20-30%; in this case the percentage of occupied hormone receptors reduced from 11.8 to 8.6%. The affinity of receptors for 1.25-(OH)2D3 did not change thereby (Kd = 0.25-0.26 nM; Kd2 = 0.06-0.10 nM). At the same time the value of cooperativity coefficient showed a decrease-from 1.7 to 1.4, which was accompanied by a reduction of the maximum capacity of receptors (1.2-1.5-fold). Vitamin C depletion augmented the manifestation of vitamin D deficiency in guinea pigs and impeded their correction after administration of cholecalciferol. This markedly retarded the restoration of the 25-OH D level in the blood as well as the number of occupied and unoccupied nuclear receptors for 1.25-(OH)2D3. The experimental results illustrate the effects of ascorbic acid on the vitamin D hormonal system function, which is manifested both at the level of 1.25-(OH)2D3 synthesis in the kidneys and of its receptor binding in target tissues.

pubmed23n0099_458

Percutaneous balloon valvuloplasty and coronary angioplasty for the treatment of calcific aortic stenosis and obstructive coronary artery disease in an elderly patient.

An 81-year-old man with severe calcific aortic stenosis and coronary artery disease who refused surgical therapy was treated with sequential percutaneous balloon aortic valvuloplasty (PBAV) and percutaneous transluminal angioplasty. Before percutaneous balloon valvuloplasty, the mean aortic gradient was 76 mmHg, and the aortic valve area was .45 cm2. The aortic valve was dilated using 15-mm and 18-mm balloons. The mean gradient decreased to 40 mmHG, and the aortic valve area increased to .62 cm2. Percutaneous transluminal coronary angioplasty (PTCA) was performed 2 weeks later, and an 85% proximal left circumflex stenosis was successfully dilated to 20%. No complications were noted during either procedure. At 6-month follow-up, the patient had returned to normal activities and was asymptomatic. Thus, combined therapy with PBAV and PTCA is technically feasible in selected elderly patients with calcific aortic stenosis and anatomically suitable coronary artery disease. This nonsurgical therapeutic approach may be useful in the treatment of selected patients who refuse or who are deferred from cardiac surgery.

Percutaneous balloon valvuloplasty and coronary angioplasty for the treatment of calcific aortic stenosis and obstructive coronary artery disease in an elderly patient. An 81-year-old man with severe calcific aortic stenosis and coronary artery disease who refused surgical therapy was treated with sequential percutaneous balloon aortic valvuloplasty (PBAV) and percutaneous transluminal angioplasty. Before percutaneous balloon valvuloplasty, the mean aortic gradient was 76 mmHg, and the aortic valve area was .45 cm2. The aortic valve was dilated using 15-mm and 18-mm balloons. The mean gradient decreased to 40 mmHG, and the aortic valve area increased to .62 cm2. Percutaneous transluminal coronary angioplasty (PTCA) was performed 2 weeks later, and an 85% proximal left circumflex stenosis was successfully dilated to 20%. No complications were noted during either procedure. At 6-month follow-up, the patient had returned to normal activities and was asymptomatic. Thus, combined therapy with PBAV and PTCA is technically feasible in selected elderly patients with calcific aortic stenosis and anatomically suitable coronary artery disease. This nonsurgical therapeutic approach may be useful in the treatment of selected patients who refuse or who are deferred from cardiac surgery.

pubmed23n0410_2314

Prognostic factors in patients with hepatocellular carcinoma treated by transcatheter arterial embolization.

Transcatheter arterial embolization induces marked antitumor response in patients with hepatocellular carcinoma, but the survival benefit of transcatheter arterial embolization remains to be determined. This study investigated prognostic factors in patients with advanced hepatocellular carcinoma treated by transcatheter arterial embolization. A total of 128 consecutive patients with non-resectable hepatocellular carcinoma, who had undergone transcatheter arterial embolization between May 1990 and August 1998, were analyzed to investigate prognostic factors. Median survival time and survival proportions at 1, 3 and 5 years were 3.3 years, 92.0, 54.6 and 23.4%, respectively. By multivariate analysis using the accelerated failure time model, age &lt;60 years, hepatitis C virus antibody positivity, serum albumin &gt;3.5 g/dl, absence of portal vein invasion and serum alpha-fetoprotein level &lt;400 ng/ml were significantly associated with favorable survival. For clinical application, we also propose a prognostic equation with combination of specific prognostic factors, by which survival curves of each patient could be predicted directly. The findings of the current study may be helpful in predicting the life expectancy of hepatocellular carcinoma patients treated by transcatheter arterial embolization and in designing future clinical trials of transcatheter arterial embolization for hepatocellular carcinoma.

Prognostic factors in patients with hepatocellular carcinoma treated by transcatheter arterial embolization. Transcatheter arterial embolization induces marked antitumor response in patients with hepatocellular carcinoma, but the survival benefit of transcatheter arterial embolization remains to be determined. This study investigated prognostic factors in patients with advanced hepatocellular carcinoma treated by transcatheter arterial embolization. A total of 128 consecutive patients with non-resectable hepatocellular carcinoma, who had undergone transcatheter arterial embolization between May 1990 and August 1998, were analyzed to investigate prognostic factors. Median survival time and survival proportions at 1, 3 and 5 years were 3.3 years, 92.0, 54.6 and 23.4%, respectively. By multivariate analysis using the accelerated failure time model, age &lt;60 years, hepatitis C virus antibody positivity, serum albumin &gt;3.5 g/dl, absence of portal vein invasion and serum alpha-fetoprotein level &lt;400 ng/ml were significantly associated with favorable survival. For clinical application, we also propose a prognostic equation with combination of specific prognostic factors, by which survival curves of each patient could be predicted directly. The findings of the current study may be helpful in predicting the life expectancy of hepatocellular carcinoma patients treated by transcatheter arterial embolization and in designing future clinical trials of transcatheter arterial embolization for hepatocellular carcinoma.

pubmed23n0509_17926

The importance of accounting for the uncertainty of published prognostic model estimates.

Reported is the importance of properly reflecting uncertainty associated with prognostic model estimates when calculating the survival benefit of a treatment or technology, using liver transplantation as an example. Monte Carlo simulation techniques were used to account for the uncertainty of prognostic model estimates using the standard errors of the regression coefficients and their correlations. These methods were applied to patients with primary biliary cirrhosis undergoing liver transplantation using a prognostic model from a historic cohort who did not undergo transplantation. The survival gain over 4 years from transplantation was estimated. Ignoring the uncertainty in the prognostic model, the estimated survival benefit of liver transplantation was 16.7 months (95 percent confidence interval [CI], 13.5 to 20.1), and was statistically significant (p &lt; .001). After adjusting for model uncertainty using the standard errors of the regression coefficients, the estimated survival benefit was 17.5 months (95 percent CI, -3.9 to 38.5) and was no longer statistically significant. An additional adjustment for the correlation between regression coefficients widened the 95 percent confidence interval slightly: the estimated survival benefit was 17.0 months (95 percent CI: -4.6 to 38.6). It is important that the precision of regression coefficients is available for users of published prognostic models. Ignoring this additional information substantially underestimates uncertainty, which can then impact misleadingly on policy decisions.

The importance of accounting for the uncertainty of published prognostic model estimates. Reported is the importance of properly reflecting uncertainty associated with prognostic model estimates when calculating the survival benefit of a treatment or technology, using liver transplantation as an example. Monte Carlo simulation techniques were used to account for the uncertainty of prognostic model estimates using the standard errors of the regression coefficients and their correlations. These methods were applied to patients with primary biliary cirrhosis undergoing liver transplantation using a prognostic model from a historic cohort who did not undergo transplantation. The survival gain over 4 years from transplantation was estimated. Ignoring the uncertainty in the prognostic model, the estimated survival benefit of liver transplantation was 16.7 months (95 percent confidence interval [CI], 13.5 to 20.1), and was statistically significant (p &lt; .001). After adjusting for model uncertainty using the standard errors of the regression coefficients, the estimated survival benefit was 17.5 months (95 percent CI, -3.9 to 38.5) and was no longer statistically significant. An additional adjustment for the correlation between regression coefficients widened the 95 percent confidence interval slightly: the estimated survival benefit was 17.0 months (95 percent CI: -4.6 to 38.6). It is important that the precision of regression coefficients is available for users of published prognostic models. Ignoring this additional information substantially underestimates uncertainty, which can then impact misleadingly on policy decisions.

pubmed23n0977_1551

CRISPR-Cas9 for cancer therapy: Opportunities and challenges.

Cancer is a genetic disease stemming from cumulative genetic/epigenetic aberrations. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9-mediated genome editing technology has been extensively applied in various cell types and organisms, both in vitro and in vivo, for efficient gene disruption and gene modification. CRISPR-Cas9 has shown great promise for the treatment of cancer. However, despite its advantages and tremendous potential, numerous challenges, such as fitness of edited cells, editing efficiency, delivery methods and potential off-target effects, remain to be solved for completely clinical application. Here, we present the potential applications and recent advances of CRISPR-Cas9 in cancer therapy, and discuss the challenges that might be encountered in clinical applications.

CRISPR-Cas9 for cancer therapy: Opportunities and challenges. Cancer is a genetic disease stemming from cumulative genetic/epigenetic aberrations. Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-Cas9-mediated genome editing technology has been extensively applied in various cell types and organisms, both in vitro and in vivo, for efficient gene disruption and gene modification. CRISPR-Cas9 has shown great promise for the treatment of cancer. However, despite its advantages and tremendous potential, numerous challenges, such as fitness of edited cells, editing efficiency, delivery methods and potential off-target effects, remain to be solved for completely clinical application. Here, we present the potential applications and recent advances of CRISPR-Cas9 in cancer therapy, and discuss the challenges that might be encountered in clinical applications.

pubmed23n0727_19885

From genes to social communication: molecular sensing by the vomeronasal organ.

The ability to distinguish molecular cues emitted by other individuals is a fundamental feature of social interactions such as finding and identifying a mate, establishing social hierarchies, and initiating interspecies defensive behaviors. In rodents, this ability involves the vomeronasal organ (VNO), a distinct chemoreceptive structure that is part of the olfactory system. Recent insights have led to unprecedented progress in identifying ligand and receptor families underlying vomeronasal recognition, characterizing the behavioral consequences caused by VNO activation, and defining higher neural circuits underlying the initiation of instinctive behaviors such as aggression. Here, we review such findings and discuss future areas for investigation, including large-scale mapping studies, immune system-VNO interactions, in vivo recording of neural activity, and optogenetic alteration of sexual and social behaviors.

From genes to social communication: molecular sensing by the vomeronasal organ. The ability to distinguish molecular cues emitted by other individuals is a fundamental feature of social interactions such as finding and identifying a mate, establishing social hierarchies, and initiating interspecies defensive behaviors. In rodents, this ability involves the vomeronasal organ (VNO), a distinct chemoreceptive structure that is part of the olfactory system. Recent insights have led to unprecedented progress in identifying ligand and receptor families underlying vomeronasal recognition, characterizing the behavioral consequences caused by VNO activation, and defining higher neural circuits underlying the initiation of instinctive behaviors such as aggression. Here, we review such findings and discuss future areas for investigation, including large-scale mapping studies, immune system-VNO interactions, in vivo recording of neural activity, and optogenetic alteration of sexual and social behaviors.

pubmed23n1021_868

Strategies to improve the implementation of healthy eating, physical activity and obesity prevention policies, practices or programmes within childcare services.

Despite the existence of effective interventions and best-practice guideline recommendations for childcare services to implement evidence-based policies, practices and programmes to promote child healthy eating, physical activity and prevent unhealthy weight gain, many services fail to do so. The primary aim of the review was to examine the effectiveness of strategies aimed at improving the implementation of policies, practices or programmes by childcare services that promote child healthy eating, physical activity and/or obesity prevention. The secondary aims of the review were to: 1. Examine the cost or cost-effectiveness of such strategies; 2. Examine any adverse effects of such strategies on childcare services, service staff or children; 3. Examine the effect of such strategies on child diet, physical activity or weight status. 4. Describe the acceptability, adoption, penetration, sustainability and appropriateness of such implementation strategies. We searched the following electronic databases on February 22 2019: Cochrane Central Register of Controlled trials (CENTRAL), MEDLINE, MEDLINE In Process, Embase, PsycINFO, ERIC, CINAHL and SCOPUS for relevant studies. We searched reference lists of included studies, handsearched two international implementation science journals, the World Health Organization International Clinical Trials Registry Platform (www.who.int/ictrp/) and ClinicalTrials.gov (www.clinicaltrials.gov). We included any study (randomised or nonrandomised) with a parallel control group that compared any strategy to improve the implementation of a healthy eating, physical activity or obesity prevention policy, practice or programme by staff of centre-based childcare services to no intervention, 'usual' practice or an alternative strategy. Centre-based childcare services included preschools, nurseries, long daycare services and kindergartens catering for children prior to compulsory schooling (typically up to the age of five to six years). Two review authors independently screened study titles and abstracts, extracted study data and assessed risk of bias; we resolved discrepancies via consensus. We performed meta-analysis using a random-effects model where studies with suitable data and homogeneity were identified; otherwise, findings were described narratively. Twenty-one studies, including 16 randomised and five nonrandomised, were included in the review. The studies sought to improve the implementation of policies, practices or programmes targeting healthy eating (six studies), physical activity (three studies) or both healthy eating and physical activity (12 studies). Studies were conducted in the United States (n = 12), Australia (n = 8) and Ireland (n = 1). Collectively, the 21 studies included a total of 1945 childcare services examining a range of implementation strategies including educational materials, educational meetings, audit and feedback, opinion leaders, small incentives or grants, educational outreach visits or academic detailing, reminders and tailored interventions. Most studies (n = 19) examined implementation strategies versus usual practice or minimal support control, and two compared alternative implementation strategies. For implementation outcomes, six studies (one RCT) were judged to be at high risk of bias overall. The review findings suggest that implementation strategies probably improve the implementation of policies, practices or programmes that promote child healthy eating, physical activity and/or obesity prevention in childcare services. Of the 19 studies that compared a strategy to usual practice or minimal support control, 11 studies (nine RCTs) used score-based measures of implementation (e.g. childcare service nutrition environment score). Nine of these studies were included in pooled analysis, which found an improvement in implementation outcomes (SMD 0.49; 95% CI 0.19 to 0.79; participants = 495; moderate-certainty evidence). Ten studies (seven RCTs) used dichotomous measures of implementation (e.g. proportion of childcare services implementing a policy or specific practice), with seven of these included in pooled analysis (OR 1.83; 95% CI 0.81 to 4.11; participants = 391; low-certainty evidence). Findings suggest that such interventions probably lead to little or no difference in child physical activity (four RCTs; moderate-certainty evidence) or weight status (three RCTs; moderate-certainty evidence), and may lead to little or no difference in child diet (two RCTs; low-certainty evidence). None of the studies reported the cost or cost-effectiveness of the intervention. Three studies assessed the adverse effects of the intervention on childcare service staff, children and parents, with all studies suggesting they have little to no difference in adverse effects (e.g. child injury) between groups (three RCTs; low-certainty evidence). Inconsistent quality of the evidence was identified across review outcomes and study designs, ranging from very low to moderate. The primary limitation of the review was the lack of conventional terminology in implementation science, which may have resulted in potentially relevant studies failing to be identified based on the search terms used. Current research suggests that implementation strategies probably improve the implementation of policies, practices or programmes by childcare services, and may have little or no effect on measures of adverse effects. However such strategies appear to have little to no impact on measures of child diet, physical activity or weight status.

Strategies to improve the implementation of healthy eating, physical activity and obesity prevention policies, practices or programmes within childcare services. Despite the existence of effective interventions and best-practice guideline recommendations for childcare services to implement evidence-based policies, practices and programmes to promote child healthy eating, physical activity and prevent unhealthy weight gain, many services fail to do so. The primary aim of the review was to examine the effectiveness of strategies aimed at improving the implementation of policies, practices or programmes by childcare services that promote child healthy eating, physical activity and/or obesity prevention. The secondary aims of the review were to: 1. Examine the cost or cost-effectiveness of such strategies; 2. Examine any adverse effects of such strategies on childcare services, service staff or children; 3. Examine the effect of such strategies on child diet, physical activity or weight status. 4. Describe the acceptability, adoption, penetration, sustainability and appropriateness of such implementation strategies. We searched the following electronic databases on February 22 2019: Cochrane Central Register of Controlled trials (CENTRAL), MEDLINE, MEDLINE In Process, Embase, PsycINFO, ERIC, CINAHL and SCOPUS for relevant studies. We searched reference lists of included studies, handsearched two international implementation science journals, the World Health Organization International Clinical Trials Registry Platform (www.who.int/ictrp/) and ClinicalTrials.gov (www.clinicaltrials.gov). We included any study (randomised or nonrandomised) with a parallel control group that compared any strategy to improve the implementation of a healthy eating, physical activity or obesity prevention policy, practice or programme by staff of centre-based childcare services to no intervention, 'usual' practice or an alternative strategy. Centre-based childcare services included preschools, nurseries, long daycare services and kindergartens catering for children prior to compulsory schooling (typically up to the age of five to six years). Two review authors independently screened study titles and abstracts, extracted study data and assessed risk of bias; we resolved discrepancies via consensus. We performed meta-analysis using a random-effects model where studies with suitable data and homogeneity were identified; otherwise, findings were described narratively. Twenty-one studies, including 16 randomised and five nonrandomised, were included in the review. The studies sought to improve the implementation of policies, practices or programmes targeting healthy eating (six studies), physical activity (three studies) or both healthy eating and physical activity (12 studies). Studies were conducted in the United States (n = 12), Australia (n = 8) and Ireland (n = 1). Collectively, the 21 studies included a total of 1945 childcare services examining a range of implementation strategies including educational materials, educational meetings, audit and feedback, opinion leaders, small incentives or grants, educational outreach visits or academic detailing, reminders and tailored interventions. Most studies (n = 19) examined implementation strategies versus usual practice or minimal support control, and two compared alternative implementation strategies. For implementation outcomes, six studies (one RCT) were judged to be at high risk of bias overall. The review findings suggest that implementation strategies probably improve the implementation of policies, practices or programmes that promote child healthy eating, physical activity and/or obesity prevention in childcare services. Of the 19 studies that compared a strategy to usual practice or minimal support control, 11 studies (nine RCTs) used score-based measures of implementation (e.g. childcare service nutrition environment score). Nine of these studies were included in pooled analysis, which found an improvement in implementation outcomes (SMD 0.49; 95% CI 0.19 to 0.79; participants = 495; moderate-certainty evidence). Ten studies (seven RCTs) used dichotomous measures of implementation (e.g. proportion of childcare services implementing a policy or specific practice), with seven of these included in pooled analysis (OR 1.83; 95% CI 0.81 to 4.11; participants = 391; low-certainty evidence). Findings suggest that such interventions probably lead to little or no difference in child physical activity (four RCTs; moderate-certainty evidence) or weight status (three RCTs; moderate-certainty evidence), and may lead to little or no difference in child diet (two RCTs; low-certainty evidence). None of the studies reported the cost or cost-effectiveness of the intervention. Three studies assessed the adverse effects of the intervention on childcare service staff, children and parents, with all studies suggesting they have little to no difference in adverse effects (e.g. child injury) between groups (three RCTs; low-certainty evidence). Inconsistent quality of the evidence was identified across review outcomes and study designs, ranging from very low to moderate. The primary limitation of the review was the lack of conventional terminology in implementation science, which may have resulted in potentially relevant studies failing to be identified based on the search terms used. Current research suggests that implementation strategies probably improve the implementation of policies, practices or programmes by childcare services, and may have little or no effect on measures of adverse effects. However such strategies appear to have little to no impact on measures of child diet, physical activity or weight status.

pubmed23n0110_16770

Immunohistochemical staining of non-Hodgkin's lymphoma in paraffin sections using the MB1 and MT1 monoclonal antibodies.

We have performed a single blind trial to assess the value of the monoclonal antibodies MB1 and MT1 in lymphoma classification. Sixty cases of non-Hodgkin's lymphoma (NHL) were stained with MB1 and MT1 using an indirect immunoperoxidase technique in paraffin sections. The majority of B tumours (27/33) stained with MB1, and most of the T tumours (24/27) stained with MT1. The MB1 antibody often produced rather weak staining but it was apparently highly specific for B cells, with only three (3/27) of the T tumours (two cases of 'malignant histiocytosis' of the intestine (MHI) and one pleomorphic T-cell lymphoma) displaying 'false' positivity. The MT1 antibody generally produced very strong staining, but it was not very selective, with 14/33 of the B lymphomas displaying 'false' positivity. the cross-reactivity observed in 17 cases led to only three misdiagnoses, two B tumours being designated as T lymphomas and one T tumour being designated as a B lymphoma. In a few cases (7/17), dual staining with both antibodies precluded firm diagnosis. In other cases (6/17), classification was possible despite some of the tumour cells showing dual staining. The seventeenth case was a plasmacytoma displaying MT1 positivity only. While the monoclonal antibodies MB1 and MT1 are of use in classifying lymphomas in paraffin section, they are not entirely lineage-specific, and the uncritical use of these two reagents alone may give rise to misdiagnosis; the use of a panel of monoclonal antibodies may yield more accurate results. As with any immunohistochemical marker, their limitations should be recognized; interpretation must be judicious and always in the context of the histological appearances.

Immunohistochemical staining of non-Hodgkin's lymphoma in paraffin sections using the MB1 and MT1 monoclonal antibodies. We have performed a single blind trial to assess the value of the monoclonal antibodies MB1 and MT1 in lymphoma classification. Sixty cases of non-Hodgkin's lymphoma (NHL) were stained with MB1 and MT1 using an indirect immunoperoxidase technique in paraffin sections. The majority of B tumours (27/33) stained with MB1, and most of the T tumours (24/27) stained with MT1. The MB1 antibody often produced rather weak staining but it was apparently highly specific for B cells, with only three (3/27) of the T tumours (two cases of 'malignant histiocytosis' of the intestine (MHI) and one pleomorphic T-cell lymphoma) displaying 'false' positivity. The MT1 antibody generally produced very strong staining, but it was not very selective, with 14/33 of the B lymphomas displaying 'false' positivity. the cross-reactivity observed in 17 cases led to only three misdiagnoses, two B tumours being designated as T lymphomas and one T tumour being designated as a B lymphoma. In a few cases (7/17), dual staining with both antibodies precluded firm diagnosis. In other cases (6/17), classification was possible despite some of the tumour cells showing dual staining. The seventeenth case was a plasmacytoma displaying MT1 positivity only. While the monoclonal antibodies MB1 and MT1 are of use in classifying lymphomas in paraffin section, they are not entirely lineage-specific, and the uncritical use of these two reagents alone may give rise to misdiagnosis; the use of a panel of monoclonal antibodies may yield more accurate results. As with any immunohistochemical marker, their limitations should be recognized; interpretation must be judicious and always in the context of the histological appearances.

pubmed23n0992_16124

Implications of exosomes as diagnostic and therapeutic strategies in cancer.

Exosomes offer a new perspective on the biology of cancer with both diagnostic and therapeutic concepts. Due to the cell-to-cell association, exosomes are involved in the progression, metastasis, and therapeutic efficacy of the tumor. They can be isolated from blood and other body fluids to determine the disease progression in the body, including cancer growth. In addition to being reservoirs of biochemical markers of cancer, exomes can be designed to restore tumor immunity. Tumor exosomes interact with different cells in the tumor microenvironment to confer beneficial modulations, responsible for stromal activity, angiogenesis, increased vascular permeability, and immune evasion. Exosomes also contribute to the metastasis with the aim of epithelial transmission to the mesenchyme and the formation of premetastatic niches. Moreover, exosomes protect cells against the cytotoxic effects of chemotherapeutic drugs and prevent the transmission of chemotherapy resistance to adjacent cells. Therefore, exosomes are essential for many fatal cancer agents, and understanding their origins and role in cancer is important. In this article, we attempted to clarify the potential of exosomes for the application in cancer diagnosis and therapy.

Implications of exosomes as diagnostic and therapeutic strategies in cancer. Exosomes offer a new perspective on the biology of cancer with both diagnostic and therapeutic concepts. Due to the cell-to-cell association, exosomes are involved in the progression, metastasis, and therapeutic efficacy of the tumor. They can be isolated from blood and other body fluids to determine the disease progression in the body, including cancer growth. In addition to being reservoirs of biochemical markers of cancer, exomes can be designed to restore tumor immunity. Tumor exosomes interact with different cells in the tumor microenvironment to confer beneficial modulations, responsible for stromal activity, angiogenesis, increased vascular permeability, and immune evasion. Exosomes also contribute to the metastasis with the aim of epithelial transmission to the mesenchyme and the formation of premetastatic niches. Moreover, exosomes protect cells against the cytotoxic effects of chemotherapeutic drugs and prevent the transmission of chemotherapy resistance to adjacent cells. Therefore, exosomes are essential for many fatal cancer agents, and understanding their origins and role in cancer is important. In this article, we attempted to clarify the potential of exosomes for the application in cancer diagnosis and therapy.

pubmed23n0039_7715

A comparison of the uptake of [75Se]selenite, [75Se]selenomethionine and [35S]methionine by tissues of ewes and lambs.

The fate of selenium, given as Na2(75)SeO3, or [75Se]selenomethionine, and of [35S]methionine administered intravenously to ewes and lambs, has been examined. The main intention was to follow the incorporation of selenium into protein in a number of tissues, including liver and kidney, and to measure the extent of that incorporation of selenoamino acid, particularly with respect to the administration of selenite. The ewes chosen were lactating ewes with lambs at foot, and the lambs were animals which had been weaned on to fodder low in selenium and were recovering from white muscle disease with selenium therapy. These two experimental situations were chosen as they offered conditions under which selenium incorporation might be considered to be maximal. Entry of isotope into milk was rapid and was greater when 75Se was given as the selenoamino acid than as selenite. In both ewes and lambs greater amounts of activity, derived from selenite, were bound to plasma proteins than to the proteins of milk. This was particularly evident in samples taken some hours after administration. This ability of the plasma to bind selenium was demonstrated by alkaline dialysis. Small, though significant amounts of selenium, derived from Na2(75)SeO3, were incorporated as selenoamino acids into the proteins of liver, kidney and pancreas, as well as into the proteins of milk and plasma. In ewes, both selenomethionine and selenocystine were identified chromatographically in enzyme digests of defatted liver and kidney. Some differences occurred in the distribution of labelled compounds in organs from lactating ewes and recovering lambs. The incorporation of selenium into protein is discussed briefly in relation to the recent findings of an association between selenium and the enzyme glutathione peroxidase.

A comparison of the uptake of [75Se]selenite, [75Se]selenomethionine and [35S]methionine by tissues of ewes and lambs. The fate of selenium, given as Na2(75)SeO3, or [75Se]selenomethionine, and of [35S]methionine administered intravenously to ewes and lambs, has been examined. The main intention was to follow the incorporation of selenium into protein in a number of tissues, including liver and kidney, and to measure the extent of that incorporation of selenoamino acid, particularly with respect to the administration of selenite. The ewes chosen were lactating ewes with lambs at foot, and the lambs were animals which had been weaned on to fodder low in selenium and were recovering from white muscle disease with selenium therapy. These two experimental situations were chosen as they offered conditions under which selenium incorporation might be considered to be maximal. Entry of isotope into milk was rapid and was greater when 75Se was given as the selenoamino acid than as selenite. In both ewes and lambs greater amounts of activity, derived from selenite, were bound to plasma proteins than to the proteins of milk. This was particularly evident in samples taken some hours after administration. This ability of the plasma to bind selenium was demonstrated by alkaline dialysis. Small, though significant amounts of selenium, derived from Na2(75)SeO3, were incorporated as selenoamino acids into the proteins of liver, kidney and pancreas, as well as into the proteins of milk and plasma. In ewes, both selenomethionine and selenocystine were identified chromatographically in enzyme digests of defatted liver and kidney. Some differences occurred in the distribution of labelled compounds in organs from lactating ewes and recovering lambs. The incorporation of selenium into protein is discussed briefly in relation to the recent findings of an association between selenium and the enzyme glutathione peroxidase.

pubmed23n0039_544

Hematologic changes in mice during and after exposure to severe hypobaric hypoxia.

Exposing mice to an atmospheric pressure of 300 mm Hg for 16 d caused a variety of hematologic effects. Hematocrit increased rapidly in the first 8 d of exposure and slowly in the second 8 d. Reticulocyte counts rose above normal, peaked on Day 8, and then fell rapidly toward the control level. Macrocytic erythrocytes, formed during exposure, remained macrocytic after the termination of exposure and after the loss of their reticulum. The posthypoxic mice proved sensitive for erythropoietin bioassay. Mice injected with normal dog serum showed a significantly higher incorporation of 59Fe than control mice injected with physiologic saline. A reduction of the duration of exposure to 10 d resulted in only a slight decrease in the sensitivity of the mouse bioassay system. However, a 16-d exposure at a pressure of 360 mm Hg resulted in considerably less sensitive bioassay animals.

Hematologic changes in mice during and after exposure to severe hypobaric hypoxia. Exposing mice to an atmospheric pressure of 300 mm Hg for 16 d caused a variety of hematologic effects. Hematocrit increased rapidly in the first 8 d of exposure and slowly in the second 8 d. Reticulocyte counts rose above normal, peaked on Day 8, and then fell rapidly toward the control level. Macrocytic erythrocytes, formed during exposure, remained macrocytic after the termination of exposure and after the loss of their reticulum. The posthypoxic mice proved sensitive for erythropoietin bioassay. Mice injected with normal dog serum showed a significantly higher incorporation of 59Fe than control mice injected with physiologic saline. A reduction of the duration of exposure to 10 d resulted in only a slight decrease in the sensitivity of the mouse bioassay system. However, a 16-d exposure at a pressure of 360 mm Hg resulted in considerably less sensitive bioassay animals.

pubmed23n0026_3951

Detection of IgG antibodies against pigeon intestinal mucosa antigens in pigeon breeders' sera using the immunofluorescent technique.

The serum of 102 pigeon breeders was examined for circulating IgG antibodies against antigens localized in pigeon intestine by the indirect immunofluorescent technique. 15 symptomatic breeders with a positive response to inhalation challenge had antibodies against mucosa tissue. The majority of asymptomatic breeders demonstrated either a fluorescence of intestinal excrements combined with a weak fluorescence of mucosa tissue or had no detectable antibodies in their serum. By the indirect immunofluorescence, antibodies were found more frequently than by immunoprecipitin reactions in agar.

Detection of IgG antibodies against pigeon intestinal mucosa antigens in pigeon breeders' sera using the immunofluorescent technique. The serum of 102 pigeon breeders was examined for circulating IgG antibodies against antigens localized in pigeon intestine by the indirect immunofluorescent technique. 15 symptomatic breeders with a positive response to inhalation challenge had antibodies against mucosa tissue. The majority of asymptomatic breeders demonstrated either a fluorescence of intestinal excrements combined with a weak fluorescence of mucosa tissue or had no detectable antibodies in their serum. By the indirect immunofluorescence, antibodies were found more frequently than by immunoprecipitin reactions in agar.

pubmed23n0835_14169

Pneumocystis pneumonia in HIV patients: a diagnostic challenge till date.

HIV has become a major health problem in India, patients commonly succumb to opportunistic infections (OIs), respiratory infections being an important cause of morbidity and their accurate diagnosis is still a challenge. Our aim was to study the occurrence of Pneumocystis pneumonia (PCP) in HIV/AIDS patients with respiratory complaints attending ART clinic and to compare various diagnostic methodologies. One hundred and twenty five HIV/AIDS patients presenting with respiratory symptoms like cough, fever, breathlessness etc, were enrolled, and induced sputum samples were collected. Samples were homogenized using glass beads and Dithiothretol. Smears were prepared and examined by Immunoflourescent staining (IFAT), Gomori methanamine silver staining (GMSS), Toludine blue O staining (TBO) and Giemsa staining for Pneumocystis jiroveci. Among the 125 patients who presented with respiratory complaints, 34 cases (27.2%) were diagnosed as having PCP. All 34 cases were detected by IFAT followed by GMSS, Giemsa and Toludine blue O staining in decreasing order. The mean CD4 count was 67.27cells/μl. PCP has become an important health problem in HIV/AIDS patients with low CD4 counts in India. IFAT remains the most sensitive method for the detection of this uncultivable organism. In resource poor settings where an immunoflourecent microscope is not available, diagnosis of PCP still remains problematic.

Pneumocystis pneumonia in HIV patients: a diagnostic challenge till date. HIV has become a major health problem in India, patients commonly succumb to opportunistic infections (OIs), respiratory infections being an important cause of morbidity and their accurate diagnosis is still a challenge. Our aim was to study the occurrence of Pneumocystis pneumonia (PCP) in HIV/AIDS patients with respiratory complaints attending ART clinic and to compare various diagnostic methodologies. One hundred and twenty five HIV/AIDS patients presenting with respiratory symptoms like cough, fever, breathlessness etc, were enrolled, and induced sputum samples were collected. Samples were homogenized using glass beads and Dithiothretol. Smears were prepared and examined by Immunoflourescent staining (IFAT), Gomori methanamine silver staining (GMSS), Toludine blue O staining (TBO) and Giemsa staining for Pneumocystis jiroveci. Among the 125 patients who presented with respiratory complaints, 34 cases (27.2%) were diagnosed as having PCP. All 34 cases were detected by IFAT followed by GMSS, Giemsa and Toludine blue O staining in decreasing order. The mean CD4 count was 67.27cells/μl. PCP has become an important health problem in HIV/AIDS patients with low CD4 counts in India. IFAT remains the most sensitive method for the detection of this uncultivable organism. In resource poor settings where an immunoflourecent microscope is not available, diagnosis of PCP still remains problematic.

pubmed23n0834_6306

Dalbavancin: A Review in Acute Bacterial Skin and Skin Structure Infections.

Intravenous dalbavancin (Dalvance™; Xydalba™) is approved for use in adult patients with acute bacterial skin and skin structure infections (ABSSSI), with the recommended regimen being a 1000 mg dose followed 1 week later by a 500 mg dose. In the multinational DISCOVER 1 and 2 trials in adult patients with ABSSSI, dalbavancin treatment was noninferior to vancomycin (for ≥ 3 days with an option to switch to oral linezolid to complete a 10- to 14-day course) in terms of early clinical success rates (assessed 48-72 h after initiation of treatment; primary endpoint required by the FDA to assess noninferiority in registration trials of ABSSSI). Clinical response rates were also similar in both treatment groups at the end of treatment (day 14-15), irrespective of geographic region or baseline characteristics, including by infection type, diabetes mellitus status, systemic inflammatory response syndrome status, causative pathogen and renal function. Dalbavancin was generally well tolerated, with adverse events generally being of mild to moderate intensity and transient. With its broad spectrum of activity against clinically relevant Gram-positive pathogens and its favourable pharmacokinetic profile that permits a convenient two-dose, once-weekly regimen with no requirement for therapeutic drug monitoring, dalbavancin is a promising emerging option for the treatment of ABSSSI in adult patients.

Dalbavancin: A Review in Acute Bacterial Skin and Skin Structure Infections. Intravenous dalbavancin (Dalvance™; Xydalba™) is approved for use in adult patients with acute bacterial skin and skin structure infections (ABSSSI), with the recommended regimen being a 1000 mg dose followed 1 week later by a 500 mg dose. In the multinational DISCOVER 1 and 2 trials in adult patients with ABSSSI, dalbavancin treatment was noninferior to vancomycin (for ≥ 3 days with an option to switch to oral linezolid to complete a 10- to 14-day course) in terms of early clinical success rates (assessed 48-72 h after initiation of treatment; primary endpoint required by the FDA to assess noninferiority in registration trials of ABSSSI). Clinical response rates were also similar in both treatment groups at the end of treatment (day 14-15), irrespective of geographic region or baseline characteristics, including by infection type, diabetes mellitus status, systemic inflammatory response syndrome status, causative pathogen and renal function. Dalbavancin was generally well tolerated, with adverse events generally being of mild to moderate intensity and transient. With its broad spectrum of activity against clinically relevant Gram-positive pathogens and its favourable pharmacokinetic profile that permits a convenient two-dose, once-weekly regimen with no requirement for therapeutic drug monitoring, dalbavancin is a promising emerging option for the treatment of ABSSSI in adult patients.

pubmed23n0985_24605

Development and evaluation of taste masked dry syrup formulation of potassium chloride.

Potassium chloride (KCl) syrup is widely used for the oral treatment of the hypokalemia. However, it is associated with unacceptable taste. In the present study, we sought to develop a palatable and easy to reconstitute KCl dry syrup as a commercially viable alternative to currently available KCl syrup. We explored the potential of Eudragit E100 as a taste-masking polymer to coat and improve the palatability of the KCl. With the help of fluid bed processor, KCl was coated with the solution containing varying amounts of Eudragit E100 (4, 6, 10 and 15%). Coating with 10% polymer solution enabled optimal fluid bed processing, higher entrapment of the KCl (81%) and better in vitro release profile in 0.1 N HCl and pH 6.8 phosphate buffer. A dry syrup formulation containing Eudragit E100 coated KCl with good physical and chemical stability in dry and reconstituted state was developed. The palatability of the optimized formulation and commercially available KCl syrup was evaluated using the Electronic Taste Sensing Machine. The developed formulation showed~ 2-fold better taste-masking compared to the commercial KCl syrup. Thus, present investigation describes the development of an effective alternative to the current KCl syrup that can offer better palatability, stability and patient compliance.

Development and evaluation of taste masked dry syrup formulation of potassium chloride. Potassium chloride (KCl) syrup is widely used for the oral treatment of the hypokalemia. However, it is associated with unacceptable taste. In the present study, we sought to develop a palatable and easy to reconstitute KCl dry syrup as a commercially viable alternative to currently available KCl syrup. We explored the potential of Eudragit E100 as a taste-masking polymer to coat and improve the palatability of the KCl. With the help of fluid bed processor, KCl was coated with the solution containing varying amounts of Eudragit E100 (4, 6, 10 and 15%). Coating with 10% polymer solution enabled optimal fluid bed processing, higher entrapment of the KCl (81%) and better in vitro release profile in 0.1 N HCl and pH 6.8 phosphate buffer. A dry syrup formulation containing Eudragit E100 coated KCl with good physical and chemical stability in dry and reconstituted state was developed. The palatability of the optimized formulation and commercially available KCl syrup was evaluated using the Electronic Taste Sensing Machine. The developed formulation showed~ 2-fold better taste-masking compared to the commercial KCl syrup. Thus, present investigation describes the development of an effective alternative to the current KCl syrup that can offer better palatability, stability and patient compliance.

pubmed23n0885_21081

Bmp5 Regulates Neural Crest Cell Survival and Proliferation via Two Different Signaling Pathways.

Neural crest progenitor cells, which give rise to many ectodermal and mesodermal derivatives, must maintain a delicate balance of apoptosis and proliferation for their final tissue contributions. Here we show that zebrafish bmp5 is expressed in neural crest progenitor cells and that it activates the Smad and Erk signaling pathways to regulate cell survival and proliferation, respectively. Loss-of-function analysis showed that Bmp5 was required for cell survival and this response is mediated by the Smad-Msxb signaling cascade. However, the Bmp5-Smad-Msxb signaling pathway had no effect on cell proliferation. In contrast, Bmp5 was sufficient to induce cell proliferation through the Mek-Erk-Id3 signaling cascade, whereas disruption of this signaling cascade had no effect on cell survival. Taken together, our results demonstrate an important regulatory mechanism for bone morphogenic protein-initiated signal transduction underlying the formation of neural crest progenitors. Stem Cells 2017;35:1003-1014.

Bmp5 Regulates Neural Crest Cell Survival and Proliferation via Two Different Signaling Pathways. Neural crest progenitor cells, which give rise to many ectodermal and mesodermal derivatives, must maintain a delicate balance of apoptosis and proliferation for their final tissue contributions. Here we show that zebrafish bmp5 is expressed in neural crest progenitor cells and that it activates the Smad and Erk signaling pathways to regulate cell survival and proliferation, respectively. Loss-of-function analysis showed that Bmp5 was required for cell survival and this response is mediated by the Smad-Msxb signaling cascade. However, the Bmp5-Smad-Msxb signaling pathway had no effect on cell proliferation. In contrast, Bmp5 was sufficient to induce cell proliferation through the Mek-Erk-Id3 signaling cascade, whereas disruption of this signaling cascade had no effect on cell survival. Taken together, our results demonstrate an important regulatory mechanism for bone morphogenic protein-initiated signal transduction underlying the formation of neural crest progenitors. Stem Cells 2017;35:1003-1014.

pubmed23n0010_4074

Mucosal temperature rises following long-term use of full dentures.

Mucosal temperatures were studied in twenty-two subjects, who had worn full upper and lower dentures for at least 5 years. By comparing with a group of twenty-two fully dentate subjects, it was found that in the denture wearers, temperatures were increased significantly in the lower jaw, but not in the upper jaw.

Mucosal temperature rises following long-term use of full dentures. Mucosal temperatures were studied in twenty-two subjects, who had worn full upper and lower dentures for at least 5 years. By comparing with a group of twenty-two fully dentate subjects, it was found that in the denture wearers, temperatures were increased significantly in the lower jaw, but not in the upper jaw.

pubmed23n0811_10070

Specific activation of dendritic cells enhances clearance of Bacillus anthracis following infection.

Dendritic cells are potent activators of the immune system and have a key role in linking innate and adaptive immune responses. In the current study we have used ex vivo pulsed bone marrow dendritic cells (BMDC) in a novel adoptive transfer strategy to protect against challenge with Bacillus anthracis, in a murine model. Pre-pulsing murine BMDC with either recombinant Protective Antigen (PA) or CpG significantly upregulated expression of the activation markers CD40, CD80, CD86 and MHC-II. Passive transfusion of mice with pulsed BMDC, concurrently with active immunisation with rPA in alum, significantly enhanced (p&lt;0.001) PA-specific splenocyte responses seven days post-immunisation. Parallel studies using ex vivo DCs expanded from human peripheral blood and activated under the same conditions as the murine DC, demonstrated that human DCs had a PA dose-related significant increase in the markers CD40, CD80 and CCR7 and that the increases in CD40 and CD80 were maintained when the other activating components, CpG and HK B. anthracis were added to the rPA in culture. Mice vaccinated on a single occasion intra-muscularly with rPA and alum and concurrently transfused intra-dermally with pulsed BMDC, demonstrated 100% survival following lethal B. anthracis challenge and had significantly enhanced (p&lt;0.05) bacterial clearance within 2 days, compared with mice vaccinated with rPA and alum alone.

Specific activation of dendritic cells enhances clearance of Bacillus anthracis following infection. Dendritic cells are potent activators of the immune system and have a key role in linking innate and adaptive immune responses. In the current study we have used ex vivo pulsed bone marrow dendritic cells (BMDC) in a novel adoptive transfer strategy to protect against challenge with Bacillus anthracis, in a murine model. Pre-pulsing murine BMDC with either recombinant Protective Antigen (PA) or CpG significantly upregulated expression of the activation markers CD40, CD80, CD86 and MHC-II. Passive transfusion of mice with pulsed BMDC, concurrently with active immunisation with rPA in alum, significantly enhanced (p&lt;0.001) PA-specific splenocyte responses seven days post-immunisation. Parallel studies using ex vivo DCs expanded from human peripheral blood and activated under the same conditions as the murine DC, demonstrated that human DCs had a PA dose-related significant increase in the markers CD40, CD80 and CCR7 and that the increases in CD40 and CD80 were maintained when the other activating components, CpG and HK B. anthracis were added to the rPA in culture. Mice vaccinated on a single occasion intra-muscularly with rPA and alum and concurrently transfused intra-dermally with pulsed BMDC, demonstrated 100% survival following lethal B. anthracis challenge and had significantly enhanced (p&lt;0.05) bacterial clearance within 2 days, compared with mice vaccinated with rPA and alum alone.

pubmed23n0109_1071

Infectious crystalline keratopathy.

Crystalline deposits developed in the anterior third of the stroma in a 60-year-old woman. The deposits resolved only after aggressive treatment with intravenously given penicillin and topical erythromycin and vancomycin hydrochloride. Review of reported cases indicated that infectious crystalline keratopathy is caused by chronic colonization of the stroma by bacteria, usually streptococci of the viridans group. Local tissue trauma, concomitant use of topical corticosteroids, an intact overlying epithelium and use of a bandage-type soft contact lens are factors in the development of the infection. Patients with crystalline formations in this setting should undergo early lamellar biopsy for histologic examination, culture and sensitivity testing, followed by aggressive therapy with appropriate antibiotics.

Infectious crystalline keratopathy. Crystalline deposits developed in the anterior third of the stroma in a 60-year-old woman. The deposits resolved only after aggressive treatment with intravenously given penicillin and topical erythromycin and vancomycin hydrochloride. Review of reported cases indicated that infectious crystalline keratopathy is caused by chronic colonization of the stroma by bacteria, usually streptococci of the viridans group. Local tissue trauma, concomitant use of topical corticosteroids, an intact overlying epithelium and use of a bandage-type soft contact lens are factors in the development of the infection. Patients with crystalline formations in this setting should undergo early lamellar biopsy for histologic examination, culture and sensitivity testing, followed by aggressive therapy with appropriate antibiotics.

pubmed23n0956_5067

Linking ecological health to co-occurring organic and inorganic chemical stressors in a groundwater-fed stream system.

Freshwaters are among the most endangered ecosystems worldwide, due predominantly to excessive anthropogenic practices compromising the future provisioning of ecosystem services. Despite increased awareness of the role of multiple stressors in accounting for ecological degradation in mixed land-use stream systems, risk assessment approaches applicable in field settings are still required. This study provides a first indication for ecological consequences of the interaction of organic and inorganic chemical stressors, not typically evaluated together, which may provide a missing link enabling the reconnection of chemical and ecological findings. Specifically, impaired ecological conditions - represented by lower abundance of meiobenthic individuals - were observed in the hyporheic zone where a contaminant groundwater plume discharged to the stream. These zones were characterized by high xenobiotic organic concentrations, and strongly reduced groundwater (e.g. elevated dissolved iron and arsenic) linked to the dissolution of iron hydroxides (iron reduction) caused by the degradation of xenobiotic compounds in the plume. Further research is still needed to separate whether impact is driven by a combined effect of organic and inorganic stressors impacting the ecological communities, or whether the conditions - when present simultaneously - are responsible for enabling a specific chemical stressor's availability (e.g. trace metals), and thus toxicity, along the study stream. Regardless, these findings suggest that benthic meioinvertebrates are promising indicators for supporting biological assessments of stream systems to sufficiently represent impacts resulting from the co-occurrence of stressors in different stream compartments. Importantly, identification of the governing circumstances is crucial for revealing key patterns and impact drivers that may be needed in correctly prioritizing stressor impacts in these systems. This study further highlights the importance of stream-aquifer interfaces for investigating chemical stressor effects in multiple stressor systems. This will require holistic approaches for linking contaminant hydrogeology and eco(toxico)logy in order to positively influence the sustainable management of water resources globally.

Linking ecological health to co-occurring organic and inorganic chemical stressors in a groundwater-fed stream system. Freshwaters are among the most endangered ecosystems worldwide, due predominantly to excessive anthropogenic practices compromising the future provisioning of ecosystem services. Despite increased awareness of the role of multiple stressors in accounting for ecological degradation in mixed land-use stream systems, risk assessment approaches applicable in field settings are still required. This study provides a first indication for ecological consequences of the interaction of organic and inorganic chemical stressors, not typically evaluated together, which may provide a missing link enabling the reconnection of chemical and ecological findings. Specifically, impaired ecological conditions - represented by lower abundance of meiobenthic individuals - were observed in the hyporheic zone where a contaminant groundwater plume discharged to the stream. These zones were characterized by high xenobiotic organic concentrations, and strongly reduced groundwater (e.g. elevated dissolved iron and arsenic) linked to the dissolution of iron hydroxides (iron reduction) caused by the degradation of xenobiotic compounds in the plume. Further research is still needed to separate whether impact is driven by a combined effect of organic and inorganic stressors impacting the ecological communities, or whether the conditions - when present simultaneously - are responsible for enabling a specific chemical stressor's availability (e.g. trace metals), and thus toxicity, along the study stream. Regardless, these findings suggest that benthic meioinvertebrates are promising indicators for supporting biological assessments of stream systems to sufficiently represent impacts resulting from the co-occurrence of stressors in different stream compartments. Importantly, identification of the governing circumstances is crucial for revealing key patterns and impact drivers that may be needed in correctly prioritizing stressor impacts in these systems. This study further highlights the importance of stream-aquifer interfaces for investigating chemical stressor effects in multiple stressor systems. This will require holistic approaches for linking contaminant hydrogeology and eco(toxico)logy in order to positively influence the sustainable management of water resources globally.

pubmed23n0266_2550

The St. Jude Medical valve. Experience with 1,000 cases.

We analyzed the long-term results of valve replacement with the St. Jude Medical bileaflet valve (St. Jude Medical, Inc., St. Paul, Minn.) in our first 1000 implantations between 1978 and 1992. A total of 399 patients had mitral valve replacement, 471 aortic valve, and 130 double (mitral and aortic) valve replacement. The average patient age was 64 +/- 15 years and the majority of patients (52%) had concomitant coronary disease. With 4328 patient-years of follow-up, 83% of the mitral group, 76% of the aortic group, and 77% of the double valve group were free of thromboembolism at 10 years after operation, and 87% of the mitral group, 82% of the aortic group, and 85% of the double valve group were free of valve-related hemorrhage. At 10 years, 91% of the mitral group, 84% of the aortic group, and 84% of the double valve group were free of valve-related death. However, overall survival at 10 years was only 42% +/- 4% for the mitral group, 43% +/- 4% for the aortic group, and 43% +/- 6% for the double valve group. For all three groups, age was a highly significant factor stratifying survival (p &lt; 0.001), as was the presence of coronary disease (all p &lt; 0.001). The excellent freedom from valve-related death at 10 years of 84% to 91% is in striking contrast to the overall survivals of 42% to 43% at 10 years. This difference suggests that the primary factors limiting long-term survival after valve replacement with the St. Jude Medical valve are not valve-related factors, but other patient factors such as age and concomitant coronary disease.

The St. Jude Medical valve. Experience with 1,000 cases. We analyzed the long-term results of valve replacement with the St. Jude Medical bileaflet valve (St. Jude Medical, Inc., St. Paul, Minn.) in our first 1000 implantations between 1978 and 1992. A total of 399 patients had mitral valve replacement, 471 aortic valve, and 130 double (mitral and aortic) valve replacement. The average patient age was 64 +/- 15 years and the majority of patients (52%) had concomitant coronary disease. With 4328 patient-years of follow-up, 83% of the mitral group, 76% of the aortic group, and 77% of the double valve group were free of thromboembolism at 10 years after operation, and 87% of the mitral group, 82% of the aortic group, and 85% of the double valve group were free of valve-related hemorrhage. At 10 years, 91% of the mitral group, 84% of the aortic group, and 84% of the double valve group were free of valve-related death. However, overall survival at 10 years was only 42% +/- 4% for the mitral group, 43% +/- 4% for the aortic group, and 43% +/- 6% for the double valve group. For all three groups, age was a highly significant factor stratifying survival (p &lt; 0.001), as was the presence of coronary disease (all p &lt; 0.001). The excellent freedom from valve-related death at 10 years of 84% to 91% is in striking contrast to the overall survivals of 42% to 43% at 10 years. This difference suggests that the primary factors limiting long-term survival after valve replacement with the St. Jude Medical valve are not valve-related factors, but other patient factors such as age and concomitant coronary disease.

pubmed23n0212_4565

The effectiveness of inactivated vaccines applied parenterally to sows to control Escherichia coli diarrhea in piglets in an industrial fattening farm.

Different vaccines against Escherichia coli diarrhea of piglets were applied parenterally in pregnant sows at an industrial fattening farm. The following vaccines were used: vaccine No. 1 with non-complete Freund's adjuvant. Tween 80 and Arlacel A, comprising O149:K91,K88; O139:K82; O8:K87,K88; O141:K85,K88; and O64:K? E. coli serotypes; vaccine No. 2 with paraffin oil instead of Freund's adjuvant, comprising the same E. coli serotypes as the vaccine No. 1; stable specific vaccine with 10% aluminum hydroxide, based on E. coli serotypes most frequently isolated from piglets which died at the farm (O149:K91,K88; O8:K87,K88; O20:K17; O64:K?); Gletvax K88 (Wellcome) and NOBI-VAC LT-K88 (Intervet International). The number of piglets which died up to the moment of weaning in comparison to the number of born ones was considered as an indicator of acquired protection. It was found that the most effective in conferring protection against E. coli diarrhea were: vaccine No. 1 and NOBI-VAC. The differences in the mortality rate between piglets originating from sows vaccinated with these vaccines and those from unvaccinated ones were statistically significant (P less than 0.05). No significant differences were noted between controls and animals vaccinated with the remaining vaccines.

The effectiveness of inactivated vaccines applied parenterally to sows to control Escherichia coli diarrhea in piglets in an industrial fattening farm. Different vaccines against Escherichia coli diarrhea of piglets were applied parenterally in pregnant sows at an industrial fattening farm. The following vaccines were used: vaccine No. 1 with non-complete Freund's adjuvant. Tween 80 and Arlacel A, comprising O149:K91,K88; O139:K82; O8:K87,K88; O141:K85,K88; and O64:K? E. coli serotypes; vaccine No. 2 with paraffin oil instead of Freund's adjuvant, comprising the same E. coli serotypes as the vaccine No. 1; stable specific vaccine with 10% aluminum hydroxide, based on E. coli serotypes most frequently isolated from piglets which died at the farm (O149:K91,K88; O8:K87,K88; O20:K17; O64:K?); Gletvax K88 (Wellcome) and NOBI-VAC LT-K88 (Intervet International). The number of piglets which died up to the moment of weaning in comparison to the number of born ones was considered as an indicator of acquired protection. It was found that the most effective in conferring protection against E. coli diarrhea were: vaccine No. 1 and NOBI-VAC. The differences in the mortality rate between piglets originating from sows vaccinated with these vaccines and those from unvaccinated ones were statistically significant (P less than 0.05). No significant differences were noted between controls and animals vaccinated with the remaining vaccines.

pubmed23n0661_13681

Inhibition of Plasmodium sporozoites infection by targeting the host cell.

There is a great need of new drugs against malaria because of the increasing spread of parasite resistance against the most commonly used drugs in the field. We found that monensin, a common veterinary antibiotic, has a strong inhibitory effect in Plasmodium berghei and Plasmodium yoelii sporozoites hepatocyte infection in vitro. Infection of host cells by another apicomplexan parasite with a similar mechanism of host cell invasion, Toxoplasma tachyzoites, was also inhibited. Treatment of mice with monensin abrogates liver infection with P. berghei sporozoites in vivo. We also found that at low concentrations monensin inhibits the infection of Plasmodium sporozoites by rendering host cells resistant to infection, rather than having a direct effect on sporozoites. Monensin effect is targeted to the initial stages of parasite invasion of the host cell with little or no effect on development, suggesting that this antibiotic affects an essential host cell component that is required for Plasmodium sporozoite invasion.

Inhibition of Plasmodium sporozoites infection by targeting the host cell. There is a great need of new drugs against malaria because of the increasing spread of parasite resistance against the most commonly used drugs in the field. We found that monensin, a common veterinary antibiotic, has a strong inhibitory effect in Plasmodium berghei and Plasmodium yoelii sporozoites hepatocyte infection in vitro. Infection of host cells by another apicomplexan parasite with a similar mechanism of host cell invasion, Toxoplasma tachyzoites, was also inhibited. Treatment of mice with monensin abrogates liver infection with P. berghei sporozoites in vivo. We also found that at low concentrations monensin inhibits the infection of Plasmodium sporozoites by rendering host cells resistant to infection, rather than having a direct effect on sporozoites. Monensin effect is targeted to the initial stages of parasite invasion of the host cell with little or no effect on development, suggesting that this antibiotic affects an essential host cell component that is required for Plasmodium sporozoite invasion.

pubmed23n0327_1242

Accumulation of rifampicin by Escherichia coli and Staphylococcus aureus.

A centrifugation method was used to investigate the accumulation of 14C-rifampicin by Staphylococcus aureus and Escherichia coli, and to characterize the mechanism of rifampicin transport into S. aureus. For both species, drug accumulation was rapid with the steady-state concentration (SSC) reached within 40 s of drug exposure. Rifampicin accumulation by S. aureus was temperature and pH dependent; the lower the experimental temperature and the lower the experimental pH, the lower was the concentration of rifampicin accumulated. Accumulation was unaffected by the presence of inhibitors of antibiotic efflux, carbonyl cyanide m-chlorophenylhydrazone (CCCP), dinitrophenol (DNP), or reserpine. Exposure to increasing concentrations of rifampicin suggested that the accumulation process was saturable above a rifampicin concentration of 0.2 mg/L. Michaelis-Menten kinetics gave an apparent Km and Vmax for rifampicin, calculated from a Lineweaver-Burk plot, of 0.05 mg/L (0.06 microM) and 3.8 ng rifampicin per second, respectively. However, calculations suggest that these values reflect those for binding of rifampicin to its target, RNA polymerase.

Accumulation of rifampicin by Escherichia coli and Staphylococcus aureus. A centrifugation method was used to investigate the accumulation of 14C-rifampicin by Staphylococcus aureus and Escherichia coli, and to characterize the mechanism of rifampicin transport into S. aureus. For both species, drug accumulation was rapid with the steady-state concentration (SSC) reached within 40 s of drug exposure. Rifampicin accumulation by S. aureus was temperature and pH dependent; the lower the experimental temperature and the lower the experimental pH, the lower was the concentration of rifampicin accumulated. Accumulation was unaffected by the presence of inhibitors of antibiotic efflux, carbonyl cyanide m-chlorophenylhydrazone (CCCP), dinitrophenol (DNP), or reserpine. Exposure to increasing concentrations of rifampicin suggested that the accumulation process was saturable above a rifampicin concentration of 0.2 mg/L. Michaelis-Menten kinetics gave an apparent Km and Vmax for rifampicin, calculated from a Lineweaver-Burk plot, of 0.05 mg/L (0.06 microM) and 3.8 ng rifampicin per second, respectively. However, calculations suggest that these values reflect those for binding of rifampicin to its target, RNA polymerase.

pubmed23n0733_4588

[Approach to perinatal mortality in the Netherlands: outcomes of a systematic expert study].

In 2009 the Minister of Health, Welfare and Sport ordered several measures to reduce the high perinatal mortality in the Netherlands. One of these was the carrying out of a descriptive study into pregnancy and birth in which the systematic consultation of experts played an important role. The main aims were to produce a list of subjects to study and to prioritize scientific research. In addition, representatives of the scientific committees of the professional groups involved and the heads of perinatology centres were also consulted. The 25 most important resulting research themes pertained to (a) early detection of risks during the preconceptional period, pregnancy and parturition, (b) recognition and tackling of societal, psychosocial, social and socio-economic risk factors, and (c) a more extensive and risk-led collaboration between all healthcare workers within the Dutch obstetric healthcare system. This study contributed to the study agenda of the Netherlands Organization for Health Research and Development (ZonMw) which resulted in a widely supported, specialty transcending research programme on all aspects of pregnancy and birth.

[Approach to perinatal mortality in the Netherlands: outcomes of a systematic expert study]. In 2009 the Minister of Health, Welfare and Sport ordered several measures to reduce the high perinatal mortality in the Netherlands. One of these was the carrying out of a descriptive study into pregnancy and birth in which the systematic consultation of experts played an important role. The main aims were to produce a list of subjects to study and to prioritize scientific research. In addition, representatives of the scientific committees of the professional groups involved and the heads of perinatology centres were also consulted. The 25 most important resulting research themes pertained to (a) early detection of risks during the preconceptional period, pregnancy and parturition, (b) recognition and tackling of societal, psychosocial, social and socio-economic risk factors, and (c) a more extensive and risk-led collaboration between all healthcare workers within the Dutch obstetric healthcare system. This study contributed to the study agenda of the Netherlands Organization for Health Research and Development (ZonMw) which resulted in a widely supported, specialty transcending research programme on all aspects of pregnancy and birth.

pubmed23n0057_15166

Protein-protein interactions of HIV-1 reverse transcriptase: implication of central and C-terminal regions in subunit binding.

Human immunodeficiency virus 1 reverse transcriptase (RT) purified from virions is composed of a approximately 51,000 Mr polypeptide and a approximately 66,000 Mr polypeptide that are thought to be in heterodimer structure (Chandra et al., 1986; Hansen et al., 1988; Starnes &amp; Cheng, 1989) and are identical except for a 15,000 Mr C-terminal truncation in the smaller species (Di Marzo-Veronese et al., 1986). We prepared individual bacterial-recombinant RTs as the approximately 66,000 Mr polypeptide (p66) or as the approximately 51,000 Mr polypeptide (p51) and then conducted various in vitro protein-protein binding experiments. Analytical ultracentrifugation studies in 0.25 M NaCl at pH 6.5 revealed that p66 was in monomer-dimer equilibrium with KA of 5.1 x 10(4) M-1. p51 failed to dimerize and behaved as a monomer under these conditions. Mixing of the p66 and p51 polypeptides resulted in a 1:1 heterodimer with KA of 4.9 x 10(5) M-1. These results on formation of the p66/p66 homodimer and p66/p51 heterodimer were confirmed by gel filtration analysis using FPLC Superose-12 columns. Binding between p66 and individual p66 segment polypeptides also was observed using an immunoprecipitation assay. Binding between p51 and p66 in this assay was resistant to the presence of approximately 1 M NaCl, suggesting that the binding free energy has a large hydrophobic component. C-Terminal truncation of p66 to yield a 29-kDa polypeptide eliminated binding to p66, and N-terminal truncation of p66 to yield a 15-kDa peptide also eliminated binding to p66. The results indicate that purified individual RT peptides p51 and p66 are capable of binding to form a 1:1 heterodimer and suggest that the central region of p66 is required for this subunit binding; the C-terminal region (15,000 Mr) of p66 appears to be required also, as p51 alone did not dimerize.

Protein-protein interactions of HIV-1 reverse transcriptase: implication of central and C-terminal regions in subunit binding. Human immunodeficiency virus 1 reverse transcriptase (RT) purified from virions is composed of a approximately 51,000 Mr polypeptide and a approximately 66,000 Mr polypeptide that are thought to be in heterodimer structure (Chandra et al., 1986; Hansen et al., 1988; Starnes &amp; Cheng, 1989) and are identical except for a 15,000 Mr C-terminal truncation in the smaller species (Di Marzo-Veronese et al., 1986). We prepared individual bacterial-recombinant RTs as the approximately 66,000 Mr polypeptide (p66) or as the approximately 51,000 Mr polypeptide (p51) and then conducted various in vitro protein-protein binding experiments. Analytical ultracentrifugation studies in 0.25 M NaCl at pH 6.5 revealed that p66 was in monomer-dimer equilibrium with KA of 5.1 x 10(4) M-1. p51 failed to dimerize and behaved as a monomer under these conditions. Mixing of the p66 and p51 polypeptides resulted in a 1:1 heterodimer with KA of 4.9 x 10(5) M-1. These results on formation of the p66/p66 homodimer and p66/p51 heterodimer were confirmed by gel filtration analysis using FPLC Superose-12 columns. Binding between p66 and individual p66 segment polypeptides also was observed using an immunoprecipitation assay. Binding between p51 and p66 in this assay was resistant to the presence of approximately 1 M NaCl, suggesting that the binding free energy has a large hydrophobic component. C-Terminal truncation of p66 to yield a 29-kDa polypeptide eliminated binding to p66, and N-terminal truncation of p66 to yield a 15-kDa peptide also eliminated binding to p66. The results indicate that purified individual RT peptides p51 and p66 are capable of binding to form a 1:1 heterodimer and suggest that the central region of p66 is required for this subunit binding; the C-terminal region (15,000 Mr) of p66 appears to be required also, as p51 alone did not dimerize.

pubmed23n0927_17285

Ouabain targets the Na⁺/K⁺-ATPase α₃ isoform to inhibit cancer cell proliferation and induce apoptosis.

Ouabain has been used for the treatment of heart failure and atrial fibrillation. Its potential anticancer effect has also attracted great interest. The aim of the present study was to evaluate the anticancer effect of ouabain and investigate its molecular target. The effects of ouabain on the viability of and induction of cellular death on OS-RC-2 renal cancer cells were examined using the MTT assay and acridine orange/ethidium bromide staining. The levels of Ca<sup2+</sup and reactive oxygen species were determined using Fura-3-acetoxymethyl ester and dichloro-dihydro-fluorescein diacetate probes, respectively. Apoptosis was examined using annexin V-fluorescein isothiocyanate/propidium iodide staining and western blotting. The expression profile of the different Na<sup+</sup/K<sup+</sup-ATPase (NKA) isoforms in NCI-H446 small cell lung cancer cells was determined using immunocytochemistry and reverse transcription polymerase chain reaction analysis. In the present study, it was demonstrated that ouabain inhibited cancer cell proliferation and induced apoptosis while no significant difference in the expression of NKA α<sub1</sub and α<sub3</sub isoforms was detected following 48 h of ouabain treatment. Furthermore, expression of NKA α<sub3</sub but not the α<sub1</sub isoform was associated with ouabain sensitivity. The results of the present study indicated that ouabain targets the NKA α<sub3</sub isoform, inhibits cancer cell proliferation and induces apoptosis.

Ouabain targets the Na⁺/K⁺-ATPase α₃ isoform to inhibit cancer cell proliferation and induce apoptosis. Ouabain has been used for the treatment of heart failure and atrial fibrillation. Its potential anticancer effect has also attracted great interest. The aim of the present study was to evaluate the anticancer effect of ouabain and investigate its molecular target. The effects of ouabain on the viability of and induction of cellular death on OS-RC-2 renal cancer cells were examined using the MTT assay and acridine orange/ethidium bromide staining. The levels of Ca<sup2+</sup and reactive oxygen species were determined using Fura-3-acetoxymethyl ester and dichloro-dihydro-fluorescein diacetate probes, respectively. Apoptosis was examined using annexin V-fluorescein isothiocyanate/propidium iodide staining and western blotting. The expression profile of the different Na<sup+</sup/K<sup+</sup-ATPase (NKA) isoforms in NCI-H446 small cell lung cancer cells was determined using immunocytochemistry and reverse transcription polymerase chain reaction analysis. In the present study, it was demonstrated that ouabain inhibited cancer cell proliferation and induced apoptosis while no significant difference in the expression of NKA α<sub1</sub and α<sub3</sub isoforms was detected following 48 h of ouabain treatment. Furthermore, expression of NKA α<sub3</sub but not the α<sub1</sub isoform was associated with ouabain sensitivity. The results of the present study indicated that ouabain targets the NKA α<sub3</sub isoform, inhibits cancer cell proliferation and induces apoptosis.

pubmed23n0967_785

Novel distillation process for effective and stable separation of high-concentration acetone-butanol-ethanol mixture from fermentation-pervaporation integration process.

One of the major obstacles of acetone-butanol-ethanol (ABE) fermentation from renewable biomass resources is the energy-intensive separation process. To decrease the energy demand of the ABE downstream separation processes, hybrid in situ separation system with conventional distillation is recognized as an effective method. However, in the distillation processes, the high reflux ratio of the ethanol column and the accumulation of ethanol on top of the water and butanol columns led to poor controllability and high operation cost of the distillations. In this study, vacuum distillation process which is based on a decanter-assisted ethanol-butanol-water recycle loop named E-TCD sequence was developed to improve the conventional separation sequence for ABE separation. The permeate of in situ pervaporation system was used as the feed. The distillation processes were simulated and optimized by iterative strategies. ABE mixture with acetone, butanol and ethanol concentrations of 115.8 g/L, 191.4 g/L and 17.8 g/L (the other composition was water) that obtained from fermentation-pervaporation integration process was used as the feed. A plant scaled to 1025 kg/h of ABE mixture was performed, and the product purities were 100 wt% of butanol, 99.7 wt% of acetone and 95 wt% of ethanol, respectively. Results showed that only 5.3 MJ/kg (of butanol) was required for ABE separation, which was only 37.54% of the energy cost in conventional distillation processes. Compared with the drawbacks of ethanol accumulation in butanol-water recycle loop and the extremely high recovery rate of ethanol in conventional distillation processes, simulation results obtained in the current work avoided the accumulation of ethanol based on the novel E-TCD sequence.

Novel distillation process for effective and stable separation of high-concentration acetone-butanol-ethanol mixture from fermentation-pervaporation integration process. One of the major obstacles of acetone-butanol-ethanol (ABE) fermentation from renewable biomass resources is the energy-intensive separation process. To decrease the energy demand of the ABE downstream separation processes, hybrid in situ separation system with conventional distillation is recognized as an effective method. However, in the distillation processes, the high reflux ratio of the ethanol column and the accumulation of ethanol on top of the water and butanol columns led to poor controllability and high operation cost of the distillations. In this study, vacuum distillation process which is based on a decanter-assisted ethanol-butanol-water recycle loop named E-TCD sequence was developed to improve the conventional separation sequence for ABE separation. The permeate of in situ pervaporation system was used as the feed. The distillation processes were simulated and optimized by iterative strategies. ABE mixture with acetone, butanol and ethanol concentrations of 115.8 g/L, 191.4 g/L and 17.8 g/L (the other composition was water) that obtained from fermentation-pervaporation integration process was used as the feed. A plant scaled to 1025 kg/h of ABE mixture was performed, and the product purities were 100 wt% of butanol, 99.7 wt% of acetone and 95 wt% of ethanol, respectively. Results showed that only 5.3 MJ/kg (of butanol) was required for ABE separation, which was only 37.54% of the energy cost in conventional distillation processes. Compared with the drawbacks of ethanol accumulation in butanol-water recycle loop and the extremely high recovery rate of ethanol in conventional distillation processes, simulation results obtained in the current work avoided the accumulation of ethanol based on the novel E-TCD sequence.

pubmed23n1095_18049

The impact of time to prostate specific antigen nadir on biochemical recurrence and mortality rates after radiation therapy for localized prostate cancer.

To investigate the effect of time to prostate-specific antigen (PSA) nadir (TTN) after radiation therapy (RTx) for prostate cancer (PCa) on biochemical recurrence (BCR) and overall survival (OS) rates. We analyzed 2,506 patients treated with RTx (external beam radiotherapy, brachytherapy or combinations) between years 2000 and 2021. Kaplan Meier and multivariable Cox regression models tested BCR-free survival and OS after stratification according to TTN (≤24 vs. 24.1-60 vs. &gt;60 months). Similar analyses were performed after stratification according to absolute PSA values at nadir (&lt;0.01 vs. 0.01-0.1 vs. 0.11-0.4 vs. &gt;0.4 ng/ml). Finally, we repeated analyses after setting the time point of PSA nadir as the beginning of follow up in survival analyses. 10-year BCR-free survival rates were 55.5, 81.7 and 91.1% and OS rates were 71.5, 79.4 and 96.1% for TTN ≤24 months, 24.1 month-60 month and &gt;60 months, respectively. Longer TTN was an independent predictor for BCR-free survival and OS (all P&lt;0.001). However, after accounting for lead-time bias, in multivariable analyses, this association remained only significant for BCR-free survival (P≤0.03), but not for OS (P≥0.1). Finally, compared to a PSA nadir of &lt;0.01 ng/ml, PSA nadir of 0.01-0.1 ng/ml, 0.11-0.4 ng/ml as well as &gt;0.4 ng/ml were independent predictors for shorter BCR-free survival (P≤0.02), but not OS (P≥0.08). Shorter time to TTN and high PSA values at nadir are indicative of early treatment failure (BCR) and OS. However, after accounting for lead-time bias, this effect only remained valid for BCR.

The impact of time to prostate specific antigen nadir on biochemical recurrence and mortality rates after radiation therapy for localized prostate cancer. To investigate the effect of time to prostate-specific antigen (PSA) nadir (TTN) after radiation therapy (RTx) for prostate cancer (PCa) on biochemical recurrence (BCR) and overall survival (OS) rates. We analyzed 2,506 patients treated with RTx (external beam radiotherapy, brachytherapy or combinations) between years 2000 and 2021. Kaplan Meier and multivariable Cox regression models tested BCR-free survival and OS after stratification according to TTN (≤24 vs. 24.1-60 vs. &gt;60 months). Similar analyses were performed after stratification according to absolute PSA values at nadir (&lt;0.01 vs. 0.01-0.1 vs. 0.11-0.4 vs. &gt;0.4 ng/ml). Finally, we repeated analyses after setting the time point of PSA nadir as the beginning of follow up in survival analyses. 10-year BCR-free survival rates were 55.5, 81.7 and 91.1% and OS rates were 71.5, 79.4 and 96.1% for TTN ≤24 months, 24.1 month-60 month and &gt;60 months, respectively. Longer TTN was an independent predictor for BCR-free survival and OS (all P&lt;0.001). However, after accounting for lead-time bias, in multivariable analyses, this association remained only significant for BCR-free survival (P≤0.03), but not for OS (P≥0.1). Finally, compared to a PSA nadir of &lt;0.01 ng/ml, PSA nadir of 0.01-0.1 ng/ml, 0.11-0.4 ng/ml as well as &gt;0.4 ng/ml were independent predictors for shorter BCR-free survival (P≤0.02), but not OS (P≥0.08). Shorter time to TTN and high PSA values at nadir are indicative of early treatment failure (BCR) and OS. However, after accounting for lead-time bias, this effect only remained valid for BCR.

pubmed23n0130_5130

Causes of glue ear. An historical review of theories and evidence.

Over the past one hundred years medical views on the cause of glue ear have frequently changed. The medical literature was reviewed to see if these changes reflected advances in the level of scientific support for different causes. This revealed that only a few of the many proposed causes command any scientific support. An explanation for the changing pattern of views on the aetiology of glue ear was therefore sought by considering secular changes in medical knowledge and belief in general. This suggested that the views held on the cause of glue ear at any given time are influenced and largely determined by the prevailing knowledge and beliefs of medicine as a whole. This phenomenon is not peculiar to glue ear--though conditions about which there is considerable uncertainty are probably more susceptible to such influences.

Causes of glue ear. An historical review of theories and evidence. Over the past one hundred years medical views on the cause of glue ear have frequently changed. The medical literature was reviewed to see if these changes reflected advances in the level of scientific support for different causes. This revealed that only a few of the many proposed causes command any scientific support. An explanation for the changing pattern of views on the aetiology of glue ear was therefore sought by considering secular changes in medical knowledge and belief in general. This suggested that the views held on the cause of glue ear at any given time are influenced and largely determined by the prevailing knowledge and beliefs of medicine as a whole. This phenomenon is not peculiar to glue ear--though conditions about which there is considerable uncertainty are probably more susceptible to such influences.

pubmed23n0809_16493

Non-anthropogenic dust exposure and asthma medication purchase in children.

Air pollution has been shown to increase frequency of asthma attacks, as usually measured by hospitalisation rates. We hypothesise that purchase of asthma reliever medications will reflect a broader association between the environmental exposure and asthma exacerbations. In a time series analysis, we estimated the association of dust storms with mild asthma manifestations, as indicated by medication purchases, during 2005-2011. We compared our results with the estimation of the association of dust storms with hospitalisations due to asthma and asthma-like symptoms. We detected 289 dust storms characterised by high levels of particulate matter &lt;10 μm in diameter. We identified 42,920 children with asthma, wheezing or asthma-like symptoms, of whom 2418 were hospitalised. We observed a higher risk of asthma medication purchase on the day of a mild dust storm (relative risk 1.05, 95% CI 1.00-1.10). The next peak in drug purchases was 3 days later and was more pronounced among Bedouin-Arab children. Stratified analyses showed higher risks for hospitalisation among Bedouin-Arab children; especially among children living in temporary houses (relative risk 1.33, 95% CI 1.04-1.71). We observed an increased risk of asthma medication purchase associated with mild dust storms. The risk observed for hospitalisation was more pronounced among the rural Bedouin-Arab population.

Non-anthropogenic dust exposure and asthma medication purchase in children. Air pollution has been shown to increase frequency of asthma attacks, as usually measured by hospitalisation rates. We hypothesise that purchase of asthma reliever medications will reflect a broader association between the environmental exposure and asthma exacerbations. In a time series analysis, we estimated the association of dust storms with mild asthma manifestations, as indicated by medication purchases, during 2005-2011. We compared our results with the estimation of the association of dust storms with hospitalisations due to asthma and asthma-like symptoms. We detected 289 dust storms characterised by high levels of particulate matter &lt;10 μm in diameter. We identified 42,920 children with asthma, wheezing or asthma-like symptoms, of whom 2418 were hospitalised. We observed a higher risk of asthma medication purchase on the day of a mild dust storm (relative risk 1.05, 95% CI 1.00-1.10). The next peak in drug purchases was 3 days later and was more pronounced among Bedouin-Arab children. Stratified analyses showed higher risks for hospitalisation among Bedouin-Arab children; especially among children living in temporary houses (relative risk 1.33, 95% CI 1.04-1.71). We observed an increased risk of asthma medication purchase associated with mild dust storms. The risk observed for hospitalisation was more pronounced among the rural Bedouin-Arab population.

pubmed23n1135_10915

Correction: Large-scale and fast synthesis of nano-hydroxyapatite powder by a microwave-hydrothermal method.

[This corrects the article DOI: 10.1039/C9RA00091G.].

Correction: Large-scale and fast synthesis of nano-hydroxyapatite powder by a microwave-hydrothermal method. [This corrects the article DOI: 10.1039/C9RA00091G.].

pubmed23n0012_5206

Dopamine uptake in serotoninergic terminals in vitro: a valuable tool for the histochemical differentiation of catecholaminergic and serotoninergic terminals in rat cerebral structures.

An in vitro method was developed to separately visualize dopaminergic, noradrenergic and serotoninergic terminals in the cerebral, hippocampal and cerebellar cortices of the rat, by means of the glyoxylic histochemical fluorescence method. Animals were pretreated with alpha-methylparatyrosine to deplete catecholamine stores. Thin vibratome sections were made and incubated in the presence of various exogenous amines and inhibitors of the catecholaminergic and serotoninergic transport systems. Experimental conditions insuring the best specificity were determined and the validity of the combined pharmacological and histochemical approach was further tested by using animals in which the cortical dopaminergic, noradrenergic or serotoninergic innervations were destroyed. Under the experimental conditions used, norepinephrine as well as alpha-methyl-norepinephrine were taken up in only noradrenergic and dopaminergic terminals. A separate visualization of the two systems was obtained by using specific uptake inhibitors. Dopamine was taken up not only in catecholaminergic but also in serotominergic terminals. The uptake of DA in serotoninergic fibers was inhibited by a specific inhibitor of the serotoninergic transport system or by the presence of serotonine in the incubating medium. The signification and the implications of thes results are discussed.

Dopamine uptake in serotoninergic terminals in vitro: a valuable tool for the histochemical differentiation of catecholaminergic and serotoninergic terminals in rat cerebral structures. An in vitro method was developed to separately visualize dopaminergic, noradrenergic and serotoninergic terminals in the cerebral, hippocampal and cerebellar cortices of the rat, by means of the glyoxylic histochemical fluorescence method. Animals were pretreated with alpha-methylparatyrosine to deplete catecholamine stores. Thin vibratome sections were made and incubated in the presence of various exogenous amines and inhibitors of the catecholaminergic and serotoninergic transport systems. Experimental conditions insuring the best specificity were determined and the validity of the combined pharmacological and histochemical approach was further tested by using animals in which the cortical dopaminergic, noradrenergic or serotoninergic innervations were destroyed. Under the experimental conditions used, norepinephrine as well as alpha-methyl-norepinephrine were taken up in only noradrenergic and dopaminergic terminals. A separate visualization of the two systems was obtained by using specific uptake inhibitors. Dopamine was taken up not only in catecholaminergic but also in serotominergic terminals. The uptake of DA in serotoninergic fibers was inhibited by a specific inhibitor of the serotoninergic transport system or by the presence of serotonine in the incubating medium. The signification and the implications of thes results are discussed.

pubmed23n0521_10811

[Children of homoparental families: psychological and sexual development].

In the last few years some studies have been made about homosexual parenthood and especially if this interferes with the child's development, identity and future sexual tendency. These studies prove that the parents' homosexuality does not seem to compromise the child's psychosexual development and that the child's possible homosexuality does not depend on the parents' sexual behaviour but on different factors linked to the relationship; as a matter of fact there are no relevant differences in the occurrence of homosexuality in children born from homosexual and heterosexual parents. The present study investigated the psychological and sexual development in a group of children of homoparental family. The sample observed included 37 children, 22 male and 15 female. The following psychodiagnostic tests were used for assessment: spontaneous drawing, draw-a-person test, family drawing, blacky pictures test. The developmental of the child's sexual identity was determined through the preferences and fantasies he/she showed while playing, in the relationship with his/her mates, in the way of dressing. We used interviews with the child and the parents. All the children showed a good adherence to their sexual role. The observations and interviews proved that the children's disorders are not linked to the parents' homosexuality. Being brought up by homosexual parents does not seem to compromise the child's future sexual tendency. However, in these children the sexual preference could change in adult life, being influenced by future situations and circumstances.

[Children of homoparental families: psychological and sexual development]. In the last few years some studies have been made about homosexual parenthood and especially if this interferes with the child's development, identity and future sexual tendency. These studies prove that the parents' homosexuality does not seem to compromise the child's psychosexual development and that the child's possible homosexuality does not depend on the parents' sexual behaviour but on different factors linked to the relationship; as a matter of fact there are no relevant differences in the occurrence of homosexuality in children born from homosexual and heterosexual parents. The present study investigated the psychological and sexual development in a group of children of homoparental family. The sample observed included 37 children, 22 male and 15 female. The following psychodiagnostic tests were used for assessment: spontaneous drawing, draw-a-person test, family drawing, blacky pictures test. The developmental of the child's sexual identity was determined through the preferences and fantasies he/she showed while playing, in the relationship with his/her mates, in the way of dressing. We used interviews with the child and the parents. All the children showed a good adherence to their sexual role. The observations and interviews proved that the children's disorders are not linked to the parents' homosexuality. Being brought up by homosexual parents does not seem to compromise the child's future sexual tendency. However, in these children the sexual preference could change in adult life, being influenced by future situations and circumstances.

pubmed23n1123_6347

Endocrine-Disrupting Chemicals and Their Adverse Effects on the Endoplasmic Reticulum.

There is growing concern regarding the health and safety issues of endocrine-disrupting chemicals (EDCs). Long-term exposure to EDCs has serious adverse health effects through both hormone-direct and hormone-indirect ways. Accordingly, some EDCs can be a pathogen and an inducer to the susceptibility of disease, even if they have a very low affinity on the estrogen receptor, or no estrogenic effect. Endoplasmic reticulum (ER) stress recently attracted attention in this research area. Because ER and ER stress could be key regulators of the EDC's adverse effects, such as the malfunction of the organ, as well as the death, apoptosis, and proliferation of a cell. In this review, we focused on finding evidence which shows that EDCs could be a trigger for ER stress and provide specific examples of EDCs, which are known to cause ER stress currently.

Endocrine-Disrupting Chemicals and Their Adverse Effects on the Endoplasmic Reticulum. There is growing concern regarding the health and safety issues of endocrine-disrupting chemicals (EDCs). Long-term exposure to EDCs has serious adverse health effects through both hormone-direct and hormone-indirect ways. Accordingly, some EDCs can be a pathogen and an inducer to the susceptibility of disease, even if they have a very low affinity on the estrogen receptor, or no estrogenic effect. Endoplasmic reticulum (ER) stress recently attracted attention in this research area. Because ER and ER stress could be key regulators of the EDC's adverse effects, such as the malfunction of the organ, as well as the death, apoptosis, and proliferation of a cell. In this review, we focused on finding evidence which shows that EDCs could be a trigger for ER stress and provide specific examples of EDCs, which are known to cause ER stress currently.

pubmed23n0383_1458

Exercise electrocardiogram testing and prognosis. Novel markers and predictive instruments.

Ample evidence now exists supporting the use of the exercise test primarily for prognostic, as opposed to diagnostic, purposes. Although limitations must be recognized, the Duke exercise treadmill score, the chronotropic response to exercise, and heart rate recovery appear to function as powerful and independent predictors of risk. With the possible exception of exercise-induced ischemia, as manifested by the ST-segment and angina components of the Duke exercise treadmill score, exercise predictors of risk are not clearly modifiable. Nonetheless, they are clinically quite useful since they may well identify patients who are or are not likely to gain benefit from further testing and aggressive therapies. How so? The "plain old" exercise treadmill test makes it possible to easily, safely, and inexpensively identify a large group of patients who are at low risk for death or major cardiac events. For this reason alone, the predictive instruments described in this article should be routinely incorporated into clinical practice. It makes no sense to perform expensive and potentially risky diagnostic tests, prescribe polypharmacy, or institute invasive therapeutic procedures in patients who are already at low risk. As an example, Weiner and colleagues found that coronary bypass grafting only benefited CASS registry patients who had a high-risk exercise test result. Future research will be needed to further refine risk stratification with the exercise test, and determine how best to use adjunctive imaging studies and to reduce risk among patients with prognostically important findings.

Exercise electrocardiogram testing and prognosis. Novel markers and predictive instruments. Ample evidence now exists supporting the use of the exercise test primarily for prognostic, as opposed to diagnostic, purposes. Although limitations must be recognized, the Duke exercise treadmill score, the chronotropic response to exercise, and heart rate recovery appear to function as powerful and independent predictors of risk. With the possible exception of exercise-induced ischemia, as manifested by the ST-segment and angina components of the Duke exercise treadmill score, exercise predictors of risk are not clearly modifiable. Nonetheless, they are clinically quite useful since they may well identify patients who are or are not likely to gain benefit from further testing and aggressive therapies. How so? The "plain old" exercise treadmill test makes it possible to easily, safely, and inexpensively identify a large group of patients who are at low risk for death or major cardiac events. For this reason alone, the predictive instruments described in this article should be routinely incorporated into clinical practice. It makes no sense to perform expensive and potentially risky diagnostic tests, prescribe polypharmacy, or institute invasive therapeutic procedures in patients who are already at low risk. As an example, Weiner and colleagues found that coronary bypass grafting only benefited CASS registry patients who had a high-risk exercise test result. Future research will be needed to further refine risk stratification with the exercise test, and determine how best to use adjunctive imaging studies and to reduce risk among patients with prognostically important findings.

pubmed23n0858_5593

Framing and the health policy process: a scoping review.

Framing research seeks to understand the forces that shape human behaviour in the policy process. It assumes that policy is a social construct and can be cast in a variety of ways to imply multiple legitimate value considerations. Frames provide the cognitive means of making sense of the social world, but discordance among them forms the basis of policy contestation. Framing, as both theory and method, has proven to generate considerable insight into the nature of policy debates in a variety of disciplines. Despite its salience for understanding health policy debates; however, little is known about the ways frames influence the health policy process. A scoping review using the Arksey and O'Malley framework was conducted. The literature on framing in the health sector was reviewed using nine health and social science databases. Articles were included that explicitly reported theory and methods used, data source(s), at least one frame, frame sponsor and evidence of a given frame's effect on the health policy process. A total of 52 articles, from 1996 to 2014, and representing 12 countries, were identified. Much of the research came from the policy studies/political science literature (n = 17) and used a constructivist epistemology. The term 'frame' was used as a label to describe a variety of ideas, packaged as values, social problems, metaphors or arguments. Frames were characterized at various levels of abstraction ranging from general ideological orientations to specific policy positions. Most articles presented multiple frames and showed how actors advocated for them in a highly contested political process. Framing is increasingly an important, yet overlooked aspect of the policy process. Further analysis on frames, framing processes and frame conflict can help researchers and policymakers to understand opaque and highly charged policy issues, which may facilitate the resolution of protracted policy controversies.

Framing and the health policy process: a scoping review. Framing research seeks to understand the forces that shape human behaviour in the policy process. It assumes that policy is a social construct and can be cast in a variety of ways to imply multiple legitimate value considerations. Frames provide the cognitive means of making sense of the social world, but discordance among them forms the basis of policy contestation. Framing, as both theory and method, has proven to generate considerable insight into the nature of policy debates in a variety of disciplines. Despite its salience for understanding health policy debates; however, little is known about the ways frames influence the health policy process. A scoping review using the Arksey and O'Malley framework was conducted. The literature on framing in the health sector was reviewed using nine health and social science databases. Articles were included that explicitly reported theory and methods used, data source(s), at least one frame, frame sponsor and evidence of a given frame's effect on the health policy process. A total of 52 articles, from 1996 to 2014, and representing 12 countries, were identified. Much of the research came from the policy studies/political science literature (n = 17) and used a constructivist epistemology. The term 'frame' was used as a label to describe a variety of ideas, packaged as values, social problems, metaphors or arguments. Frames were characterized at various levels of abstraction ranging from general ideological orientations to specific policy positions. Most articles presented multiple frames and showed how actors advocated for them in a highly contested political process. Framing is increasingly an important, yet overlooked aspect of the policy process. Further analysis on frames, framing processes and frame conflict can help researchers and policymakers to understand opaque and highly charged policy issues, which may facilitate the resolution of protracted policy controversies.

pubmed23n0073_12937

Glycoprotein specificity of cold-reactive IgM antilymphocyte autoantibodies in systemic lupus erythematosus.

Sera from patients with systemic lupus erythematosus frequently contain IgM antibodies to glycoproteins of Mr 46,000 and approximately 200,000 isolated from nonionic detergent lysates of mature T cells by affinity chromatography with solid-phase wheat germ agglutinin. Autoantibodies of this specificity correlate strongly with the presence of IgM anti-T cell autoantibodies, as determined by independent indirect immunofluorescence and complement-dependent microcytotoxicity assays, and are specifically absorbed by incubation of patient serum with viable T cells. Collectively, the data suggest that gp46 and, to a lesser extent, gp approximately 200 represent major targets of IgM antilymphocyte autoantibodies in systemic lupus erythematosus.

Glycoprotein specificity of cold-reactive IgM antilymphocyte autoantibodies in systemic lupus erythematosus. Sera from patients with systemic lupus erythematosus frequently contain IgM antibodies to glycoproteins of Mr 46,000 and approximately 200,000 isolated from nonionic detergent lysates of mature T cells by affinity chromatography with solid-phase wheat germ agglutinin. Autoantibodies of this specificity correlate strongly with the presence of IgM anti-T cell autoantibodies, as determined by independent indirect immunofluorescence and complement-dependent microcytotoxicity assays, and are specifically absorbed by incubation of patient serum with viable T cells. Collectively, the data suggest that gp46 and, to a lesser extent, gp approximately 200 represent major targets of IgM antilymphocyte autoantibodies in systemic lupus erythematosus.

pubmed23n0501_12619

The new rHuEPO alpha (epotin) in the management of anemia of end-stage renal disease in patients on maintenance hemodialysis.

Recombinant human erythropoietin has proved to be effective to treat anemia of end-stage renal disease (ESRD). The aim of this study was to assess the efficacy and safety profile of Epotin, a rHuEPO produced in the Middle East. One hundred thirty patients with Hct &lt;/= 27%; Hb &lt;/= 9 g/dL maintained on hemodialysis thrice weekly from 19centers in eight countries in the Middle East were recruited into this 13-week study. Depleted iron stores (TSTAT &lt;20% and/or Serum ferritin &lt; 100 microg/dL) were replenished prior to initiation of Epotin therapy, which was delivered intravenously in a dose of 150 U/kg body weight/week in three equal doses postdialysis and titrated according to hemoglobin (Hb) and hematocrit (Hct) response. Efficacy was assessed in terms of Hb/Hct response. Epotin raised the mean Hb level from 7.7 (+/- 1.2) g/dL to 12.0 (+/- 1.7) g/dL and Hct from 22.7 (+/- 4.1) % to 36.2 (+/- 5.7) % by week 13. The increase started to show significance at week 3. Targeting an absolute increase in Hb of 2.5 g/dL (Hct 7.5%) over a 13-week period, the success rate was of &lt;85.71%. Segregating patients into subgroups of men and women and chronic ESRD versus recent ESRD failed to reveal a significant differences in either the severity of the anemia or the response to Epotin. Side effects were similar to other erythropoietins; no dropouts were reported. In conclusion, Epotin is effective to treat anemia in patients on maintenance hemodialysis with an acceptable safety profile. No difference in response was observed between men and women, nor between patients with different levels of chronicity of ESRD.

The new rHuEPO alpha (epotin) in the management of anemia of end-stage renal disease in patients on maintenance hemodialysis. Recombinant human erythropoietin has proved to be effective to treat anemia of end-stage renal disease (ESRD). The aim of this study was to assess the efficacy and safety profile of Epotin, a rHuEPO produced in the Middle East. One hundred thirty patients with Hct &lt;/= 27%; Hb &lt;/= 9 g/dL maintained on hemodialysis thrice weekly from 19centers in eight countries in the Middle East were recruited into this 13-week study. Depleted iron stores (TSTAT &lt;20% and/or Serum ferritin &lt; 100 microg/dL) were replenished prior to initiation of Epotin therapy, which was delivered intravenously in a dose of 150 U/kg body weight/week in three equal doses postdialysis and titrated according to hemoglobin (Hb) and hematocrit (Hct) response. Efficacy was assessed in terms of Hb/Hct response. Epotin raised the mean Hb level from 7.7 (+/- 1.2) g/dL to 12.0 (+/- 1.7) g/dL and Hct from 22.7 (+/- 4.1) % to 36.2 (+/- 5.7) % by week 13. The increase started to show significance at week 3. Targeting an absolute increase in Hb of 2.5 g/dL (Hct 7.5%) over a 13-week period, the success rate was of &lt;85.71%. Segregating patients into subgroups of men and women and chronic ESRD versus recent ESRD failed to reveal a significant differences in either the severity of the anemia or the response to Epotin. Side effects were similar to other erythropoietins; no dropouts were reported. In conclusion, Epotin is effective to treat anemia in patients on maintenance hemodialysis with an acceptable safety profile. No difference in response was observed between men and women, nor between patients with different levels of chronicity of ESRD.

pubmed23n0974_3473

Electrochemical Hydrogen Evolution at the Interface of Monolayer VS₂ and Water from First-Principles Calculations.

Density functional theory calculations are carried out to study the hydrogen evolution reaction (HER) at the electrochemical double-layer interface of monolayer 2H phase VS<sub2</sub and water. Under typical conditions of HER, the catalyst surface is predicted to have a low hydrogen coverage of about 12%, whereas the aqueous solution side features a high hydronium concentration of about 8.3%. As a result, the HER takes place through the Volmer-Heyrovsky route, with an overall reaction barrier of about 1.0 eV, much larger than that of 1T phase VS<sub2</sub. This result demonstrates that 2H phase VS<sub2</sub is much less reactive than its 1T phase counterpart, and the 1T-to-2H phase transformation induced by thickness reduction may deteriorate the HER activity of VS<sub2</sub.

Electrochemical Hydrogen Evolution at the Interface of Monolayer VS₂ and Water from First-Principles Calculations. Density functional theory calculations are carried out to study the hydrogen evolution reaction (HER) at the electrochemical double-layer interface of monolayer 2H phase VS<sub2</sub and water. Under typical conditions of HER, the catalyst surface is predicted to have a low hydrogen coverage of about 12%, whereas the aqueous solution side features a high hydronium concentration of about 8.3%. As a result, the HER takes place through the Volmer-Heyrovsky route, with an overall reaction barrier of about 1.0 eV, much larger than that of 1T phase VS<sub2</sub. This result demonstrates that 2H phase VS<sub2</sub is much less reactive than its 1T phase counterpart, and the 1T-to-2H phase transformation induced by thickness reduction may deteriorate the HER activity of VS<sub2</sub.

pubmed23n0399_3781

Relationship of serum testosterone concentrations to mate preferences in rams.

This study examined systemic testosterone concentrations in rams that were classified according to their sexual behavior and partner preference as either female-oriented (FOR), male-oriented (MOR), or asexual (NOR). For this purpose, we measured testosterone concentrations under three separate conditions: in conscious rams during the nonbreeding season (June) and breeding season (November), and in anesthetized rams during the breeding season. Basal testosterone concentrations in conscious rams were not different among the three groups (P &gt; 0.05) in either season. However, when rams were anesthetized, mean systemic concentrations of testosterone in FORs (mean +/- SEM, 13.9 +/- 7.4 ng/ml serum) were greater (P &lt; 0.05) than in NORs (0.9 +/- 0.1 ng/ml), but not in MORs (2.2 +/- 6.2 ng/ml), whereas testosterone concentrations were not different between MORs and NORs (P &gt; 0.05). Concentrations of testosterone in the spermatic vein of FORs (127 +/- 66 ng/ml) were greater (P &lt; 0.05) than in MORs (41 +/- 10 ng/ml) and NORs (19 +/- 7 ng/ml). Serum LH concentrations were not different. Cortisol was higher (P &lt; 0.05) in anesthetized MORs (25.1 +/- 4.2 ng/ml) and NORs (27.2 +/- 4.4 ng/ml) than in FORs (10.9 +/- 1.8 ng/ml). These results demonstrate that circulating testosterone concentrations are related to sexual behavior only when rams are bled under anesthesia. Thus, differences in basal androgen concentrations in adulthood cannot be responsible for expression of male-oriented preferences or low libido in sheep. Instead, functional differences must exist between the brains of rams that differ in sexual preference expression.

Relationship of serum testosterone concentrations to mate preferences in rams. This study examined systemic testosterone concentrations in rams that were classified according to their sexual behavior and partner preference as either female-oriented (FOR), male-oriented (MOR), or asexual (NOR). For this purpose, we measured testosterone concentrations under three separate conditions: in conscious rams during the nonbreeding season (June) and breeding season (November), and in anesthetized rams during the breeding season. Basal testosterone concentrations in conscious rams were not different among the three groups (P &gt; 0.05) in either season. However, when rams were anesthetized, mean systemic concentrations of testosterone in FORs (mean +/- SEM, 13.9 +/- 7.4 ng/ml serum) were greater (P &lt; 0.05) than in NORs (0.9 +/- 0.1 ng/ml), but not in MORs (2.2 +/- 6.2 ng/ml), whereas testosterone concentrations were not different between MORs and NORs (P &gt; 0.05). Concentrations of testosterone in the spermatic vein of FORs (127 +/- 66 ng/ml) were greater (P &lt; 0.05) than in MORs (41 +/- 10 ng/ml) and NORs (19 +/- 7 ng/ml). Serum LH concentrations were not different. Cortisol was higher (P &lt; 0.05) in anesthetized MORs (25.1 +/- 4.2 ng/ml) and NORs (27.2 +/- 4.4 ng/ml) than in FORs (10.9 +/- 1.8 ng/ml). These results demonstrate that circulating testosterone concentrations are related to sexual behavior only when rams are bled under anesthesia. Thus, differences in basal androgen concentrations in adulthood cannot be responsible for expression of male-oriented preferences or low libido in sheep. Instead, functional differences must exist between the brains of rams that differ in sexual preference expression.

pubmed23n0090_17566

Structure-activity relationship of ligands of dihydrouracil dehydrogenase from mouse liver.

One hundred and five nucleobase analogues were screened as inhibitors of dihydrouracil dehydrogenase (DHUDase, EC 1.3.1.2) from mouse liver. 5-Benzyloxybenzyluracil, 1-deazauracil (2,6-pyridinediol), 3-deazauracil (2,4-pyridinediol), 5-benzyluracil, 5-nitrobarbituric acid and 5,6-dioxyuracil (alloxan) were identified as potent inhibitors of this activity, with apparent Ki values of 0.2, 0.5, 2.1, 3.4, 3.8 and 6.6 microM respectively. Both 5-benzyloxybenzyluracil and 1-deazauracil were also potent inhibitors of DHUDase from human livers. These findings along with an extensive review of literature allowed the formulation of a structure-activity relationship. The binding to DHUDase required intact C2 and C4 oxo groups. Replacement of N1 or N3 by an endocyclic carbon enhanced binding. In contrast, replacement of C5 or C6 by an endocyclic nitrogen abolished binding. Addition of a charged group to C5 and/or C6, and of a hydrophobic group to C5 but not C6 improved the binding.

Structure-activity relationship of ligands of dihydrouracil dehydrogenase from mouse liver. One hundred and five nucleobase analogues were screened as inhibitors of dihydrouracil dehydrogenase (DHUDase, EC 1.3.1.2) from mouse liver. 5-Benzyloxybenzyluracil, 1-deazauracil (2,6-pyridinediol), 3-deazauracil (2,4-pyridinediol), 5-benzyluracil, 5-nitrobarbituric acid and 5,6-dioxyuracil (alloxan) were identified as potent inhibitors of this activity, with apparent Ki values of 0.2, 0.5, 2.1, 3.4, 3.8 and 6.6 microM respectively. Both 5-benzyloxybenzyluracil and 1-deazauracil were also potent inhibitors of DHUDase from human livers. These findings along with an extensive review of literature allowed the formulation of a structure-activity relationship. The binding to DHUDase required intact C2 and C4 oxo groups. Replacement of N1 or N3 by an endocyclic carbon enhanced binding. In contrast, replacement of C5 or C6 by an endocyclic nitrogen abolished binding. Addition of a charged group to C5 and/or C6, and of a hydrophobic group to C5 but not C6 improved the binding.

pubmed23n1091_14608

Drug Development in Tissue-Agnostic Indications.

A better understanding of cancer biology has led to the development of targeted therapies specifically designed to modulate an altered molecular pathway in the cancer cells or their microenvironment. Despite the identification of molecular targets across cancer types, most of targeted therapies were developed per cancer type. In this ancestral paradigm, randomization was the gold-standard approach for market access. Randomization of large patient populations was feasible for drugs developed in common cancer types but more challenging in rare cancer types. The traditional paradigm of drug development in oncology was further challenged by the ever-expanding molecular segmentation of cancer with ever-smaller subgroups of patients who might benefit from specific targeted therapies or immunotherapies and the identification of molecular alterations against which drugs may be effective across cancer types. In this novel drug development paradigm, novel ways of evaluating the efficacy of drugs are highly needed in these small patient populations. One approach is to use each patient as his/her own control by comparing the efficacy of a drug to the efficacy of prior treatments received. This approach allows to overcome patient heterogeneity, especially in a tissue-agnostic drug development paradigm.

Drug Development in Tissue-Agnostic Indications. A better understanding of cancer biology has led to the development of targeted therapies specifically designed to modulate an altered molecular pathway in the cancer cells or their microenvironment. Despite the identification of molecular targets across cancer types, most of targeted therapies were developed per cancer type. In this ancestral paradigm, randomization was the gold-standard approach for market access. Randomization of large patient populations was feasible for drugs developed in common cancer types but more challenging in rare cancer types. The traditional paradigm of drug development in oncology was further challenged by the ever-expanding molecular segmentation of cancer with ever-smaller subgroups of patients who might benefit from specific targeted therapies or immunotherapies and the identification of molecular alterations against which drugs may be effective across cancer types. In this novel drug development paradigm, novel ways of evaluating the efficacy of drugs are highly needed in these small patient populations. One approach is to use each patient as his/her own control by comparing the efficacy of a drug to the efficacy of prior treatments received. This approach allows to overcome patient heterogeneity, especially in a tissue-agnostic drug development paradigm.

pubmed23n0387_20471

Three silent periods in the orbiculari oculi muscles of man: normal findings and some clinical vignettes.

To investigate how many true silent periods could be found in the orbiculari oculi muscles of man. 10 subjects, clinically healthy (5 male, 5 female), with a mean age of 34 years-old (range: 23 to 48) were evaluated by mean of the blink reflex at resting and during contraction of the orbiculari oculi reflex according to protocols validated internationally. Three responses called R1, R2 and R3 were obtained in the orbicular oculi muscle at resting state which had latencies and amplitudes within normal limits. What was new was to obtain three silent periods when the subjects were evaluated during muscle contraction. The duration of the first silent period was statistically longer than the second one (p &lt; 0.004) and shorter than the third silent period (p &lt; 0.0001). In addition, this test was found useful in detecting more specific findings in patients with hemifacial spasm and Meigge syndrome. This is by the first time that three silent periods in the orbicular oculi muscles are consistently demonstrated. The refractoriness of the alpha motoneurons and the action of gamma-collateral activity seem to be the main conditions leasing to display the first two periods of muscle suppression. The modification of gamma motoneurons firing as well as a pause of muscle spindles in facial muscles due to the action of nociceptive stimuli traveling unmyelinated C fibers of the supraorbital nerve might be the most important mechanisms involved in the production of the third silent period. These results enables further clinical application of this test.

Three silent periods in the orbiculari oculi muscles of man: normal findings and some clinical vignettes. To investigate how many true silent periods could be found in the orbiculari oculi muscles of man. 10 subjects, clinically healthy (5 male, 5 female), with a mean age of 34 years-old (range: 23 to 48) were evaluated by mean of the blink reflex at resting and during contraction of the orbiculari oculi reflex according to protocols validated internationally. Three responses called R1, R2 and R3 were obtained in the orbicular oculi muscle at resting state which had latencies and amplitudes within normal limits. What was new was to obtain three silent periods when the subjects were evaluated during muscle contraction. The duration of the first silent period was statistically longer than the second one (p &lt; 0.004) and shorter than the third silent period (p &lt; 0.0001). In addition, this test was found useful in detecting more specific findings in patients with hemifacial spasm and Meigge syndrome. This is by the first time that three silent periods in the orbicular oculi muscles are consistently demonstrated. The refractoriness of the alpha motoneurons and the action of gamma-collateral activity seem to be the main conditions leasing to display the first two periods of muscle suppression. The modification of gamma motoneurons firing as well as a pause of muscle spindles in facial muscles due to the action of nociceptive stimuli traveling unmyelinated C fibers of the supraorbital nerve might be the most important mechanisms involved in the production of the third silent period. These results enables further clinical application of this test.

pubmed23n0092_5788

Cis-acting elements of the sea urchin histone H2A modulator bind transcriptional factors.

Functional tests, performed by microinjection into Xenopus laevis oocytes, show that a DNA fragment containing the modulator of the early histone H2A gene of Paracentrotus lividus enhances transcription of a reporter gene when located, in the physiological orientation, upstream of the tk basal promoter. Gel retardation and DNase I footprinting assays further reveal that the H2A modulator contains at least two binding sites [upstream sequence elements 1 and 2 (USE 1 and USE 2)] for nuclear factors extracted from sea urchin embryos, which actively transcribe the early histone gene set. Interestingly, USE 1 is highly homologous to a cis-acting element previously identified in the H2A modulator of Psammechinus miliaris [Grosschedl, R., Mächler, M., Rohrer, U. &amp; Birnstiel, M. L. (1983) Nucleic Acids Res. 11, 8123-8136]. Finally, a cloned oligonucleotide containing the USE 1 sequence competes efficiently in Xenopus oocytes with the H2A modulator to prevent enhancement of transcription of the reporter gene. From these results, we conclude that USE 1 and perhaps USE 2 in the H2A modulator are upstream transcriptional elements that are recognized by trans-acting factors common to Xenopus and sea urchin.

Cis-acting elements of the sea urchin histone H2A modulator bind transcriptional factors. Functional tests, performed by microinjection into Xenopus laevis oocytes, show that a DNA fragment containing the modulator of the early histone H2A gene of Paracentrotus lividus enhances transcription of a reporter gene when located, in the physiological orientation, upstream of the tk basal promoter. Gel retardation and DNase I footprinting assays further reveal that the H2A modulator contains at least two binding sites [upstream sequence elements 1 and 2 (USE 1 and USE 2)] for nuclear factors extracted from sea urchin embryos, which actively transcribe the early histone gene set. Interestingly, USE 1 is highly homologous to a cis-acting element previously identified in the H2A modulator of Psammechinus miliaris [Grosschedl, R., Mächler, M., Rohrer, U. &amp; Birnstiel, M. L. (1983) Nucleic Acids Res. 11, 8123-8136]. Finally, a cloned oligonucleotide containing the USE 1 sequence competes efficiently in Xenopus oocytes with the H2A modulator to prevent enhancement of transcription of the reporter gene. From these results, we conclude that USE 1 and perhaps USE 2 in the H2A modulator are upstream transcriptional elements that are recognized by trans-acting factors common to Xenopus and sea urchin.

pubmed23n0378_19886

Behavioral treatment of insomnia: treatment outcome and the relevance of medical and psychiatric morbidity.

Recently, we undertook a case series study and found that behavior therapy for insomnia was effective as plied in the clinic setting and that the findings were similar to those in the "clinical trial" literature. In the present study, we evaluate a second set of case series data to assess (1) the replicability of our original findings, (2) if our treatment outcomes are statistically comparable to those in the literature, and (3) if medical and psychiatric morbidity influence treatment outcome. It was found that patients who completed four or more sessions of cognitive behavioral therapy for insomnia (CBT) were, on average, 33% improved. This average corresponded to a 56% reduction in wake time after sleep onset, a 34% reduction in sleep latency, a 29% increase in total sleep time, and a 13% decrease in number of awakenings per night. These findings are not significantly different from those reported in literature for both CBT and pharmacotherapy interventions. Medical and psychiatric comorbidity did not influence treatment outcome.

Behavioral treatment of insomnia: treatment outcome and the relevance of medical and psychiatric morbidity. Recently, we undertook a case series study and found that behavior therapy for insomnia was effective as plied in the clinic setting and that the findings were similar to those in the "clinical trial" literature. In the present study, we evaluate a second set of case series data to assess (1) the replicability of our original findings, (2) if our treatment outcomes are statistically comparable to those in the literature, and (3) if medical and psychiatric morbidity influence treatment outcome. It was found that patients who completed four or more sessions of cognitive behavioral therapy for insomnia (CBT) were, on average, 33% improved. This average corresponded to a 56% reduction in wake time after sleep onset, a 34% reduction in sleep latency, a 29% increase in total sleep time, and a 13% decrease in number of awakenings per night. These findings are not significantly different from those reported in literature for both CBT and pharmacotherapy interventions. Medical and psychiatric comorbidity did not influence treatment outcome.

pubmed23n0307_190

A large-conductance chloride channel in pigmented ciliary epithelial cells activated by GTPgammaS.

A large-conductance (or maxi-) chloride channel was identified in bovine pigmented ciliary epithelial (PCE) cells using inside-out excised patch clamp recording. The channel had a mean conductance of 293 pS when excised patches were bathed in symmetrical 130 mm NaCl although the conductance decreased to 209 pS when the solution bathing the cytoplasmic face of the patch contained only 33 mm NaCl. The channel was highly selective for chloride, with a PCl/PNa = 24. A flickery, reversible block was produced by the diuretic stilbene 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS), while 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) produced a permanent block. The channel was rarely active in cell-attached patches and usually required several minutes of polarization before activity could be detected in excised patches, a process known as metagenesis. Once activated, the channel was voltage-dependent and was mainly open within the voltage range -30 to +30 mV closing when the membrane was polarized to larger values. GTPgammaS (100 microM) activated the channel with a latency of 170 sec when applied to the cytoplasmic face of patches. This activation was not reversible upon return to control solution within the duration of the experiment. We assess the available evidence and suggest a role for this channel in volume regulation.

A large-conductance chloride channel in pigmented ciliary epithelial cells activated by GTPgammaS. A large-conductance (or maxi-) chloride channel was identified in bovine pigmented ciliary epithelial (PCE) cells using inside-out excised patch clamp recording. The channel had a mean conductance of 293 pS when excised patches were bathed in symmetrical 130 mm NaCl although the conductance decreased to 209 pS when the solution bathing the cytoplasmic face of the patch contained only 33 mm NaCl. The channel was highly selective for chloride, with a PCl/PNa = 24. A flickery, reversible block was produced by the diuretic stilbene 4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid (SITS), while 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) produced a permanent block. The channel was rarely active in cell-attached patches and usually required several minutes of polarization before activity could be detected in excised patches, a process known as metagenesis. Once activated, the channel was voltage-dependent and was mainly open within the voltage range -30 to +30 mV closing when the membrane was polarized to larger values. GTPgammaS (100 microM) activated the channel with a latency of 170 sec when applied to the cytoplasmic face of patches. This activation was not reversible upon return to control solution within the duration of the experiment. We assess the available evidence and suggest a role for this channel in volume regulation.

pubmed23n0983_4804

Stereotypical patterns of epileptiform calcium signal in hippocampal CA1, CA3, dentate gyrus and entorhinal cortex in freely moving mice.

Epilepsy is a multi-etiological brain dysfunction syndrome. Hippocampal neuronal damage induced by seizures may be one of the causes leading to cognitive impairment, but the underlying mechanism remains to be further elucidated. The kainic acid (KA) model of temporal lobe epilepsy is widely used in understanding of the epileptogenesis. Fiber photometry is a signal detection technology suitable for recording calcium activity of neurons in the deep brain of freely moving animal. Here, we used the optical fiber-based method to monitor the real-time neuronal population activities of freely moving mice after subcutaneous injection of KA. We observed that KA administration led to one to three kinds of stereotypical patterns of epileptiform calcium activity in CA1, CA3, and dentate gyrus (DG) of the hippocampus, as well as the entorhinal cortex (EC). There were three kinds of waves in the hippocampal CA1, which we named wave 1, wave 2 and slow flash. Wave 1 and wave 2 appeared in both the CA3 and DG regions, but the EC only showed wave 1. In these epileptiform calcium signals, we observed a high amplitude and long duration calcium wave as a part of wave 2, which resembled cortical spreading depression (CSD) and always appeared at or after the end of seizure. Because the same characteristic of epileptiform calcium signal appeared in different brain regions, calcium signal may not exist with region specificity, but may exhibit a cell type specific manner. Thus, our work provides a support for the pathogenesis of epilepsy and epileptiform signal transmission research.

Stereotypical patterns of epileptiform calcium signal in hippocampal CA1, CA3, dentate gyrus and entorhinal cortex in freely moving mice. Epilepsy is a multi-etiological brain dysfunction syndrome. Hippocampal neuronal damage induced by seizures may be one of the causes leading to cognitive impairment, but the underlying mechanism remains to be further elucidated. The kainic acid (KA) model of temporal lobe epilepsy is widely used in understanding of the epileptogenesis. Fiber photometry is a signal detection technology suitable for recording calcium activity of neurons in the deep brain of freely moving animal. Here, we used the optical fiber-based method to monitor the real-time neuronal population activities of freely moving mice after subcutaneous injection of KA. We observed that KA administration led to one to three kinds of stereotypical patterns of epileptiform calcium activity in CA1, CA3, and dentate gyrus (DG) of the hippocampus, as well as the entorhinal cortex (EC). There were three kinds of waves in the hippocampal CA1, which we named wave 1, wave 2 and slow flash. Wave 1 and wave 2 appeared in both the CA3 and DG regions, but the EC only showed wave 1. In these epileptiform calcium signals, we observed a high amplitude and long duration calcium wave as a part of wave 2, which resembled cortical spreading depression (CSD) and always appeared at or after the end of seizure. Because the same characteristic of epileptiform calcium signal appeared in different brain regions, calcium signal may not exist with region specificity, but may exhibit a cell type specific manner. Thus, our work provides a support for the pathogenesis of epilepsy and epileptiform signal transmission research.

pubmed23n0022_12154

Post meningococcal acute glomerular nephritis.

A case of meningococcal meningitis is described in which 10 days later there developed the histological lesions of acute exsudative proliferative glomerular nephritis without proteinuria, hematuria, hypertension or salt and water retention. The relationship between structural and functional changes in the kidney in glomerular nephritis is discussed in the light of these findings.

Post meningococcal acute glomerular nephritis. A case of meningococcal meningitis is described in which 10 days later there developed the histological lesions of acute exsudative proliferative glomerular nephritis without proteinuria, hematuria, hypertension or salt and water retention. The relationship between structural and functional changes in the kidney in glomerular nephritis is discussed in the light of these findings.

pubmed23n0755_2751

Possible role of Escherichia coli in propagation and perpetuation of chronic inflammation in ulcerative colitis.

This study investigated a possible role of Escherichia coli in propagation and perpetuation of the chronic inflammation in ulcerative colitis (UC). The lesions of UC are located superficially on the rectal and/or colonic mucosa. It is suggested that the commensal bacteria of the digestive tract may play a role in the pathogenesis of UC. Several studies have demonstrated proliferation of E. coli in the gut of UC patients. An increase in the number of E. coli in the inflamed tissue is most probably related to the abundance of iron ions produced by the bacteria. Colon mucosal biopsies were collected from 30 patients with acute-phase UC, both from tissues with inflammatory changes (n = 30) and unchanged tissue with no inflammatory changes (n = 30) from the same patient. Biopsies were also taken from 16 patients with irritable bowel syndrome diarrhea who comprised the control group. Quantitative and qualitative analysis of the biopsy specimens was performed using culture methods and real-time polymerase chain reaction (PCR). Genotyping of the E. coli isolates was done using pulsed-field gel electrophoresis. Multiplex PCR was used to compare the E. coli strains for the presence of genes responsible for synthesis of iron acquisition proteins: iroN, iutA, iha, ireA, chuA, and hlyA. We demonstrated that there was a significant increase in the number of E. coli at the sites of inflammation in patients with UC compared to the control group (P = 0.031). Comparative analysis of the restriction patterns of E. coli isolated from inflammatory and unchanged tissues showed that the local inflammatory changes did not promote specific E. coli strains. There was a significant difference in the frequency of the iroN gene in E. coli isolated from patients with UC as compared to the control group. The increase in the numbers of E. coli in the inflammatory tissues is related to the presence of chuA and iutA genes, which facilitate iron acquisition during chronic intestinal inflammatory processes.

Possible role of Escherichia coli in propagation and perpetuation of chronic inflammation in ulcerative colitis. This study investigated a possible role of Escherichia coli in propagation and perpetuation of the chronic inflammation in ulcerative colitis (UC). The lesions of UC are located superficially on the rectal and/or colonic mucosa. It is suggested that the commensal bacteria of the digestive tract may play a role in the pathogenesis of UC. Several studies have demonstrated proliferation of E. coli in the gut of UC patients. An increase in the number of E. coli in the inflamed tissue is most probably related to the abundance of iron ions produced by the bacteria. Colon mucosal biopsies were collected from 30 patients with acute-phase UC, both from tissues with inflammatory changes (n = 30) and unchanged tissue with no inflammatory changes (n = 30) from the same patient. Biopsies were also taken from 16 patients with irritable bowel syndrome diarrhea who comprised the control group. Quantitative and qualitative analysis of the biopsy specimens was performed using culture methods and real-time polymerase chain reaction (PCR). Genotyping of the E. coli isolates was done using pulsed-field gel electrophoresis. Multiplex PCR was used to compare the E. coli strains for the presence of genes responsible for synthesis of iron acquisition proteins: iroN, iutA, iha, ireA, chuA, and hlyA. We demonstrated that there was a significant increase in the number of E. coli at the sites of inflammation in patients with UC compared to the control group (P = 0.031). Comparative analysis of the restriction patterns of E. coli isolated from inflammatory and unchanged tissues showed that the local inflammatory changes did not promote specific E. coli strains. There was a significant difference in the frequency of the iroN gene in E. coli isolated from patients with UC as compared to the control group. The increase in the numbers of E. coli in the inflammatory tissues is related to the presence of chuA and iutA genes, which facilitate iron acquisition during chronic intestinal inflammatory processes.

pubmed23n0613_19344

Adjunctive analgesic therapy in veterinary medicine.

Adjunctive analgesic therapies are interventions for pain that involve agents or techniques other than the traditional analgesics (opioids, nonsteroidal anti-inflammatory drugs, and local anesthetics). Adjunctive therapies may be pharmacologic or nonpharmacologic in nature. The focus of this article is on pharmacologic interventions with potential utility as adjunctive analgesics in veterinary medicine. Pharmacology of selected agents, including medetomidine, ketamine, amantadine, gabapentin, systemic lidocaine, and pamidronate, is discussed in addition to evidence for their safety and efficacy and guidelines for their use in veterinary patients.

Adjunctive analgesic therapy in veterinary medicine. Adjunctive analgesic therapies are interventions for pain that involve agents or techniques other than the traditional analgesics (opioids, nonsteroidal anti-inflammatory drugs, and local anesthetics). Adjunctive therapies may be pharmacologic or nonpharmacologic in nature. The focus of this article is on pharmacologic interventions with potential utility as adjunctive analgesics in veterinary medicine. Pharmacology of selected agents, including medetomidine, ketamine, amantadine, gabapentin, systemic lidocaine, and pamidronate, is discussed in addition to evidence for their safety and efficacy and guidelines for their use in veterinary patients.

pubmed23n0353_8409

Cutaneous mucinosis and mastocytosis in a shar-pei.

A 7-year-old shar-pei was presented because of a recurrent dermatologic condition. Skin biopsies revealed an idiopathic (primary) cutaneous mucinosis that initially responded to corticosteroids. The condition reappeared 2 years later and subsequent biopsies revealed a mast cell tumor in some of the skin sites previously diagnosed with mucinosis.

Cutaneous mucinosis and mastocytosis in a shar-pei. A 7-year-old shar-pei was presented because of a recurrent dermatologic condition. Skin biopsies revealed an idiopathic (primary) cutaneous mucinosis that initially responded to corticosteroids. The condition reappeared 2 years later and subsequent biopsies revealed a mast cell tumor in some of the skin sites previously diagnosed with mucinosis.

pubmed23n0304_2732

Genetic alterations and oxidative metabolism in sporadic colorectal tumors from a Spanish community.

Deletions of loci on chromosomes 5q, 17p, 18q, and 22q, together with the incidence of p53 mutations and amplification of the double minute-2 gene were investigated in the sporadic colorectal tumors of 44 patients from a Spanish community. Chromosome deletions were analyzed by means of loss of heterozygosity analysis using a restriction fragment length polymorphism assay. Allelic losses were also detected by polymerase chain reaction (PCR)-single-stranded conformation polymorphism (SSCP) analysis of a polymorphic site in intron 2 of the p53 gene. The percentages of genetic deletions on the screened chromosomes were 39.3% (5q), 58.3% (17p), 40.9% (18q), and 40% (22q). Mutations in p53 exons 2-9 were examined by PCR-SSCP analysis and direct sequencing of the mutated region. Twenty of 44 tumor samples (45.45%) showed mutations at various exons except for exons 2, 3, and 9, the most frequent changes being G--&gt;T transversion and C--&gt;T transition. Because oxygen-free radicals play a role in the carcinogenesis process, we evaluated the oxidative status of the colorectal tumors. Antioxidant activities, lipid peroxidation, and DNA-damaged product concentrations in colon tumors and normal mucosa were compared. In tumor tissues, superoxide dismutase and catalase decreased fourfold and twofold, respectively, whereas glutathione peroxidase and reduced glutathione increased threefold. Malondialdehyde and 8-hydroxy-2-deoxyguanosine (8-OHdG) levels were twofold higher in colorectal tumors than in normal mucosa. Seven of 10 DNA tumor samples (70%) showing higher values of 8-OHdG also had genetic alterations at different chromosomal loci. In these samples, the p53 gene was deleted or mutated in 71.4% of cases. We concluded that the observed changes in the oxidative metabolism of the tumor cells and the consecutive increase in DNA damage may potentiate the genomic instability of different chromosomal regions, leading to further cell malignancy and tumor expansion.

Genetic alterations and oxidative metabolism in sporadic colorectal tumors from a Spanish community. Deletions of loci on chromosomes 5q, 17p, 18q, and 22q, together with the incidence of p53 mutations and amplification of the double minute-2 gene were investigated in the sporadic colorectal tumors of 44 patients from a Spanish community. Chromosome deletions were analyzed by means of loss of heterozygosity analysis using a restriction fragment length polymorphism assay. Allelic losses were also detected by polymerase chain reaction (PCR)-single-stranded conformation polymorphism (SSCP) analysis of a polymorphic site in intron 2 of the p53 gene. The percentages of genetic deletions on the screened chromosomes were 39.3% (5q), 58.3% (17p), 40.9% (18q), and 40% (22q). Mutations in p53 exons 2-9 were examined by PCR-SSCP analysis and direct sequencing of the mutated region. Twenty of 44 tumor samples (45.45%) showed mutations at various exons except for exons 2, 3, and 9, the most frequent changes being G--&gt;T transversion and C--&gt;T transition. Because oxygen-free radicals play a role in the carcinogenesis process, we evaluated the oxidative status of the colorectal tumors. Antioxidant activities, lipid peroxidation, and DNA-damaged product concentrations in colon tumors and normal mucosa were compared. In tumor tissues, superoxide dismutase and catalase decreased fourfold and twofold, respectively, whereas glutathione peroxidase and reduced glutathione increased threefold. Malondialdehyde and 8-hydroxy-2-deoxyguanosine (8-OHdG) levels were twofold higher in colorectal tumors than in normal mucosa. Seven of 10 DNA tumor samples (70%) showing higher values of 8-OHdG also had genetic alterations at different chromosomal loci. In these samples, the p53 gene was deleted or mutated in 71.4% of cases. We concluded that the observed changes in the oxidative metabolism of the tumor cells and the consecutive increase in DNA damage may potentiate the genomic instability of different chromosomal regions, leading to further cell malignancy and tumor expansion.

pubmed23n0944_2320

Multiple large-scale gene and genome duplications during the evolution of hexapods.

Polyploidy or whole genome duplication (WGD) is a major contributor to genome evolution and diversity. Although polyploidy is recognized as an important component of plant evolution, it is generally considered to play a relatively minor role in animal evolution. Ancient polyploidy is found in the ancestry of some animals, especially fishes, but there is little evidence for ancient WGDs in other metazoan lineages. Here we use recently published transcriptomes and genomes from more than 150 species across the insect phylogeny to investigate whether ancient WGDs occurred during the evolution of Hexapoda, the most diverse clade of animals. Using gene age distributions and phylogenomics, we found evidence for 18 ancient WGDs and six other large-scale bursts of gene duplication during insect evolution. These bursts of gene duplication occurred in the history of lineages such as the Lepidoptera, Trichoptera, and Odonata. To further corroborate the nature of these duplications, we evaluated the pattern of gene retention from putative WGDs observed in the gene age distributions. We found a relatively strong signal of convergent gene retention across many of the putative insect WGDs. Considering the phylogenetic breadth and depth of the insect phylogeny, this observation is consistent with polyploidy as we expect dosage balance to drive the parallel retention of genes. Together with recent research on plant evolution, our hexapod results suggest that genome duplications contributed to the evolution of two of the most diverse lineages of eukaryotes on Earth.

Multiple large-scale gene and genome duplications during the evolution of hexapods. Polyploidy or whole genome duplication (WGD) is a major contributor to genome evolution and diversity. Although polyploidy is recognized as an important component of plant evolution, it is generally considered to play a relatively minor role in animal evolution. Ancient polyploidy is found in the ancestry of some animals, especially fishes, but there is little evidence for ancient WGDs in other metazoan lineages. Here we use recently published transcriptomes and genomes from more than 150 species across the insect phylogeny to investigate whether ancient WGDs occurred during the evolution of Hexapoda, the most diverse clade of animals. Using gene age distributions and phylogenomics, we found evidence for 18 ancient WGDs and six other large-scale bursts of gene duplication during insect evolution. These bursts of gene duplication occurred in the history of lineages such as the Lepidoptera, Trichoptera, and Odonata. To further corroborate the nature of these duplications, we evaluated the pattern of gene retention from putative WGDs observed in the gene age distributions. We found a relatively strong signal of convergent gene retention across many of the putative insect WGDs. Considering the phylogenetic breadth and depth of the insect phylogeny, this observation is consistent with polyploidy as we expect dosage balance to drive the parallel retention of genes. Together with recent research on plant evolution, our hexapod results suggest that genome duplications contributed to the evolution of two of the most diverse lineages of eukaryotes on Earth.

pubmed23n0575_9734

Fly ash collected from electrostatic precipitators: microcrystalline structures and the mystery of the spheres.

Scanning electron micrographs demonstrate the presence of microcrystalline structures on the surface of coal-derived fly ash samples taken from electrostatic precipitator hoppers. Cenospheres (hollow spheres) were found to be packed with smaller cenospheres, which were also packed with spheres. Microspheres, apparently formed by uneven heating, are encapsulated in the parent sphere. Chemical analyses provide a basis for the postulation of a mechanism of formation for plerospheres (hollow spheres packed with spheres) and microcrystals.

Fly ash collected from electrostatic precipitators: microcrystalline structures and the mystery of the spheres. Scanning electron micrographs demonstrate the presence of microcrystalline structures on the surface of coal-derived fly ash samples taken from electrostatic precipitator hoppers. Cenospheres (hollow spheres) were found to be packed with smaller cenospheres, which were also packed with spheres. Microspheres, apparently formed by uneven heating, are encapsulated in the parent sphere. Chemical analyses provide a basis for the postulation of a mechanism of formation for plerospheres (hollow spheres packed with spheres) and microcrystals.

pubmed23n0594_18963

Stem cell biology and its therapeutic applications in the setting of spinal cord injury.

The development of an acute traumatic spinal cord injury (SCI) inevitably leads to a complex cascade of ischemia and inflammation that results in significant scar tissue formation. The development of such scar tissue provides a severe impediment to neural regeneration and healing with restoration of function. A multimodal approach to treatment is required because SCIs occur with differing levels of severity and over different lengths of time. To achieve significant breakthroughs in outcomes, such approaches must combine both neuroprotective and neuroregenerative treatments. Novel techniques modulating endogenous stem cells demonstrate great promise in promoting neuroregeneration and restoring function.

Stem cell biology and its therapeutic applications in the setting of spinal cord injury. The development of an acute traumatic spinal cord injury (SCI) inevitably leads to a complex cascade of ischemia and inflammation that results in significant scar tissue formation. The development of such scar tissue provides a severe impediment to neural regeneration and healing with restoration of function. A multimodal approach to treatment is required because SCIs occur with differing levels of severity and over different lengths of time. To achieve significant breakthroughs in outcomes, such approaches must combine both neuroprotective and neuroregenerative treatments. Novel techniques modulating endogenous stem cells demonstrate great promise in promoting neuroregeneration and restoring function.

pubmed23n0662_17760

Nurses' self-concept and perceived quality of care: a narrative analysis.

The perceptions of staff nurses on factors affecting patient care quality and safety have received little attention in the literature. Narrative analysis of comments provided by 106 staff nurses working in a medical-surgical setting revealed that nurses experienced contradictions and unmet expectations related to their professional role. The consequence was feelings of powerlessness, isolation, and low self-esteem, which affected nurses' perceived ability to provide quality patient care and ensure patient safety. This perceived inability to act in a professionally autonomous manner on behalf of patients, in turn, influenced nurses' professional self-concept. Recommendations are offered to enhance nurses' professional self-concept through staff development and policy changes.

Nurses' self-concept and perceived quality of care: a narrative analysis. The perceptions of staff nurses on factors affecting patient care quality and safety have received little attention in the literature. Narrative analysis of comments provided by 106 staff nurses working in a medical-surgical setting revealed that nurses experienced contradictions and unmet expectations related to their professional role. The consequence was feelings of powerlessness, isolation, and low self-esteem, which affected nurses' perceived ability to provide quality patient care and ensure patient safety. This perceived inability to act in a professionally autonomous manner on behalf of patients, in turn, influenced nurses' professional self-concept. Recommendations are offered to enhance nurses' professional self-concept through staff development and policy changes.