Euglena gracilis: Biochemical properties of a membrane bound ecto-phosphatase activity modulated by fluoroaluminate complexes and different trophic conditions.
The ecto-phosphatases belong to a group of enzymes closely associated with the cell surface that has its catalytic site facing the extracellular environment, where different phosphorylated substrates can be hydrolyzed. In the present work, we biochemically characterized the ecto-phosphatase activity of the freshwater microalgae Euglena gracilis, a model microorganism, ubiquitously distributed and resistant to several environmental stressors. The ecto-phosphatase activity is acidic, stimulated by copper and presents the following apparent kinetic parameters: Km = 2.52 ± 0.12 mM p-NPP and Vmax = 3.62 ± 0.06 nmol p-NP × h-1 × 106 cells. We observed that zinc, orthovanadate, molybdate, fluoride, and inorganic phosphate inhibit the ecto-phosphatase activity with different magnitudes. Fluoroaluminate complexes are also inhibitors of this ecto-phosphatase activity. They can be formed in the enzyme reaction conditions and are likely to occur in a natural environment where E. gracilis can be found. The ecto-phosphatase activity is constant through the culture growth phases and is negatively modulated after continuous subculturing in the dark when a shift from phototrophic to the heterotrophic metabolism is likely. The analysis of those biochemical parameters may contribute to understanding the role of E. gracilis ecto-phosphatase activity in natural environments.


Effects of pharmacological intervention on coagulopathy and organ function in xenoperfused kidneys.
Following pig to primate kidney transplantation, xenogenic activation of the coagulation (XAC) system of the recipient eventually leading to organ dysfunction and disseminated intravascular coagulation (DIC) can be observed. Using an ex-vivo perfusion circuit based on low-dose heparin-mediated anticoagulation and exogenous complement inhibition by C1- Inhibitor (C1-Inh), we have analysed XAC following contact of human blood with porcine endothelium. Porcine kidneys (n = 23) were recovered following in situ cold perfusion with histidine-tryptophan-ketoglutarate (HTK) solution and were connected to a perfusion circuit utilizing freshly drawn pooled human AB blood. Kidney survival during organ perfusion with human blood, CI-Inh, heparin but without any further pharmacological intervention was 126 +/- 78 min. XAC was observed with significantly elevated levels of D-dimer and thrombin antithrombin complexes (TAT). Pharmacological intervention with nitroprusside and prostacycline resulted in increased organ survival (220 +/- 28 min and 180 +/- 85 min respectively) but failed to inhibit XAC. In contrast, addition of activated protein C (APC) significantly reduced the increase in D-dimer and TAT and prolonged organ survival to 240 min (+/-0). On histology, no remarkable signs of XAC were observed. We conclude that exogenous APC is able to reduce XAC in this ex vivo perfusion model.


Chemically-tagged Mitsunobu reagents for use in solution-phase chemical library synthesis.
A general method for high-throughput product purification of Mitsunobu reactions is described. Tagged phosphine and azodicarboxylate reagents are used to synthesize individual library members in solution-phase. Workup and purification are easily accomplished by post-reaction sequestration of the tagged reagents and reagent byproducts by a complementary functionalized ion exchange resin. The reagents are utilized in a 3 step library synthesis.


Attenuation of ganglioside GM1 accumulation in the brain of GM1 gangliosidosis mice by neonatal intravenous gene transfer.
A single intravenous injection with 4 x 10(7) PFU of recombinant adenovirus encoding mouse beta-galactosidase cDNA to newborn mice provided widespread increases of beta-galactosidase activity, and attenuated the development of the disease including the brain at least for 60 days. The beta-galactosidase activity showed 2-4 times as high a normal activity in the liver and lung, and 50 times in the heart. In the brain, while the activity was only 10-20% of normal, the efficacy of the treatment was distinct. At the 30th day after the injection, significant attenuation of ganglioside GM1 accumulation in the cerebrum was shown in three out of seven mice. At the 60th day after the injection, the amount of ganglioside GM1 was above the normal range in all treated mice, which was speculated to be the result of reaccumulation. However, the values were still definitely lower in most of the treated mice than those in untreated mice. In the histopathological study, X-gal-positive cells, which showed the expression of exogenous beta-galactosidase gene, were observed in the brain. It is noteworthy that neonatal administration via blood vessels provided access to the central nervous system because of the incompletely formed blood-brain barrier.


There is no additional bactericidal efficacy of Environmental Protection Agency-registered disinfectant towelettes after surface drying or beyond label contact time.
Disinfectant towelettes are commonly used for surface disinfection to prevent health care-associated infections; however, there is limited consensus as to whether a surface needs to remain wet for the full label contact time after the disinfectant towelette has been used in order for complete efficacy to be achieved. The purpose of this study was to quantify the effect of contact time, including times before and after a product dries, on bactericidal efficacy of 6 towelette products registered by the Environmental Protection Agency . Six disinfectant towelette products were tested at varying contact times, including defined label contact times. Quantitative Environmental Protection Agency MB-33-00 was used to measure disinfectant efficacy against Staphylococcus aureus on Formica. Complete dry time for each disinfectant was measured gravimetrically. There were significant differences in dry times among the towelette products; contact time did not have a significant effect on bactericidal efficacy. There was no longitudinal effect when a disinfectant's contact time was greater than defined label contact time, irrespective of whether the product was wet or dry on the surface. Overall, bactericidal efficacy varied by towelette product tested and surface area wiped. Wiping larger surface areas may lead to decreased bactericidal efficacy but is product dependent. There was no additional bactericidal effect after a product dried, indicating that extended contact times beyond when the product dries will not enhance disinfection.


Structure determination of membrane proteins in five easy pieces.
Rotational Alignment (RA) solid-state NMR provides the basis for a general method for determining the structures of membrane proteins in phospholipid bilayers under physiological conditions. Membrane proteins are high priority targets for structure determination, and are challenging for existing experimental methods. Because membrane proteins reside in liquid crystalline phospholipid bilayer membranes it is important to study them in this type of environment. The RA solid-state NMR approach we have developed can be summarized in five steps, and incorporates methods of molecular biology, biochemistry, sample preparation, the implementation of NMR experiments, and structure calculations. It relies on solid-state NMR spectroscopy to obtain high-resolution spectra and residue-specific structural restraints for membrane proteins that undergo rotational diffusion around the membrane normal, but whose mobility is otherwise restricted by interactions with the membrane phospholipids. High resolution spectra of membrane proteins alone and in complex with other proteins and ligands set the stage for structure determination and functional studies of these proteins in their native, functional environment.


Cardioprotective effects of co-administration of thymoquinone and ischemic postconditioning in diabetic rats.
Ischemia/reperfusion (I/R) is a leading cause of myocardial infarction (MI) injury, contributing to excess injury to cardiac tissues involved in inflammation, apoptosis, and oxidative stress. The present study was conducted to examine the effects of combined thymoquinone (TQ) with ischemic postconditioning (IPostC) therapy on apoptosis and inflammation due to I/R injury in diabetic rat hearts. A single dose injection of streptozotocin (STZ; 60 mg/kg) was administered to thirty-two Wistar male rats to induce diabetes. Hearts were fixed on a Langendorff setting and exposed to a 30 min regional ischemia subsequently to 60 min reperfusion. IPostC was induced at the onset of reperfusion by 3 cycles of 30 sec R/I. ELISA, Western blotting assay, and TUNEL staining were applied to assess the cardioprotective effect of IPostC and TQ against I/R injury in diabetic and non-diabetic rats. Administration of TQ alone in non-diabetic isolated hearts significantly diminished CK-MB, TNF-α, IL-1β, and apoptosis and enhanced p-GSK-3β and Bcl-2 (P<0.05). Following administration of TQ, the cardioprotective effects of IPostC by elevating p-GSK-3β and Bcl-2 and alleviating apoptosis and inflammation were reestablished compared with non-IPostC diabetic hearts. These results provide substantial evidence that co-administration of TQ plus IPostC can exert cardioprotective effects on diabetic myocardium during I/R damage by attenuating the inflammatory response and apoptosis.


A ZO1-GFP fusion protein to study the dynamics of tight junctions in living cells.
To examine the dynamics of tight junctions (TJs) in living cells, chimera between the TJ-associated protein ZO-1 and green fluorescent protein (GFP) were constructed. If ZO-1 fused to the C-terminus of GFP (ZO1-CGFP) was stably expressed in MDCK cells, it was fully incorporated into TJs and colocalized with endogenous ZO-1. The GFP tag did not influence cell growth, transepithelial electrical resistance, and paracellular mannitol transport. The morphology of the transfected cells was unchanged. The ZO1-CGFP MDCK cell line thus represents an excellent tool to study TJ dynamics. The influence of the external calcium ion concentration on the formation and dynamics of TJs in living cells was thus explored. Upon opening of the TJs under short-term treatment with EGTA (up to 20 min), the localization of ZO1-CGFP at the membrane persisted. The rim-like pattern around the individual cells appeared fuzzier than in non-treated cells. Long-term calcium depletion resulted in the localization of ZO1-CGFP in the cytoplasm and the nucleus. After restoration of normal Ca(2+) concentrations, cell-cell contacts were restored and the localization of ZO1-CGFP was indistinguishable from the one in control cells kept at normal Ca(2+) concentrations. It remains open how the different localizations of ZO-1 correspond to changes in the signal transduction activity of the molecule.


The photoadaptive response to ultraviolet exposure in human skin using ultraviolet spectrophotometry.
Both pigmentation and non-pigmentary processes contribute to the development of photoadaptation yet the exact contribution of either in the resting state and in response to ultraviolet (UV) radiation is unclear. The purpose of this study was to estimate independently these changes occurring in the epidermis following repeated exposure to UV in two groups with differing degrees of constitutive pigmentation. We describe a mathematical model for explaining the spectral absorbance of excised human epidermis based on the absorbance of constituent chromophores. The model was applied to spectral absorbance data measured on samples of epidermis excised from pre-irradiated skin and from skin obtained following UV irradiation on 3 successive days. We found that in Asian skin there was only a mild photoadaptive response, principally by a small increase in pigmentation. On the other hand, the significant adaptive response in Caucasian skin was through hyperplasia of the epidermis, with tanning contributing only to a much smaller degree. This study has enabled us to study independently the pigmentary and non-pigmentary pathways and has shown that in those people with a lower degree of constitutive pigment, the primary mechanism of photoadaptation is via the non-pigmentary route.


A one month high fat diet disrupts the gut microbiome and integrity of the colon inducing adiposity and behavioral despair in male Sprague Dawley rats.
High-fat diet (HFD) is associated with gut microbiome dysfunction and mental disorders. However, the time-dependence as to when this occurs is unclear. We hypothesized that a short-term HFD causes colonic tissue integrity changes resulting in behavioral changes. Rats were fed HFD or low-fat diet (LFD) for a month and gut microbiome, colon, and behavior were evaluated. Behavioral despair was found in the HFD group. Although obesity was absent, the HFD group showed increased percent weight gain, epididymal fat tissue, and leptin expression. Moreover, the HFD group had increased colonic damage, decreased expression of the tight junction proteins, and higher lipopolysaccharides (LPS) in serum. Metagenomic analysis revealed that the HFD group had more Bacteroides and less S24-7 which correlated with the decreased claudin-5. Finally, HFD group showed an increase of microglia percent area, increased astrocytic projections, and decreased phospho-mTOR. In conclusion, HFD consumption in a short period is still sufficient to disrupt gut integrity resulting in LPS infiltration, alterations in the brain, and behavioral despair even in the absence of obesity.


Identification of AGT and CD44 in methotrexate-resistant colorectal cancer and reversal of methotrexate-resistance.
This study aims to screen out hub genes in 2 methotrexate-resistant colorectal cancer (CRC) cells (HT29 and Caco2), compared with parental CRC cells and reverse methotrexate-resistance in methotrexate-resistant CRC. GEO database and R software were utilized to analyze the gene expression profiles GSE11440 and GSE16066. Venn diagram was used to identify intersection differentially expressed genes (DEGs) between GSE11440 and GSE16066. Protein-protein interaction (PPI) was utilized to screen out central node genes. Hub genes were determined by volcano graphs, heatmaps and box plots. The functional enrichment analysis was exhibited with DAVID. The GEPIA was used to obtain survival curves to analyze association between patient prognosis and hub genes. Western blotting was used to detect the expressions of hub genes. CCK-8 assay was used to show MTX-resistant CRC cell viability following CD44 inhibitor (THIQ) and AGT inhibitor (O6-BG) treatments. In our results, there were 180 intersection DEGs between GSE11440 and GSE16066. CD44 and AGT were screened out as hub genes by PPI, heatmaps, volcano and box plots. In the 2 MTX-resistant CRC cells, the expressions of CD44 and AGT were up-regulated compared with parental CRC cells. The results of western blotting showed that CD44 and AGT were up-regulated in MTX-resistant HT29 and Caco2 cells compared with parental CRC cells. CCK-8 assay results showed that the combination of MTX with O6-BG or THIQ could significantly reduce the activity of MTX-resistant CRC cells. This research screened out CD44 and AGT in MTX-resistant CRC cells by bioinformatics and suggested that the combination of MTX with O6-BG or THIQ could enhance the sensitivity of MTX-resistant CRC cells to MTX. This research provides a new strategy for overcoming MTX-resistance in CRC.


A fundamental study for quantitative measurement of ultrasound contrast concentration by low mechanical index contrast ultrasonography.
In high mechanical index (MI) contrast ultrasonography it has been shown that the power of contrast signal intensity (CI) has a strong linear correlation with the concentration of the ultrasound contrast agent under conditions of constant applied acoustic pressure. However, it is unclear whether the linearity is preserved in low-MI contrast ultrasonography. Thus, we investigated the relationship between ultrasound contrast concentration and CI in vitro. Solutions of the ultrasound contrast agents Definity and Imagent were prepared at concentrations of 0.5, 2, 8, 32, and 128 μl/l. Placing a jelly block between the transducer and the solution, the solutions were transmitted using pulse subtraction imaging with an MI of 0.05, 0.1, and 0.5. CI was measured in dB in a region of interest 3 mm in height placed just below the border between the jelly and the solution. Data were plotted using double logarithm scales, where the concentration was expressed in dB as 10 × log (concentration). CI in dB had a strong linear correlation with concentration in dB for both agents with any MI. Best fitted slopes were close to 1, indicating that the power of CI is proportional to the concentration. In low-MI contrast ultrasonography, the power of CI is proportional to contrast concentration, and CI in dB is logarithmic to the concentration. Thus, the microbubble concentration can be quantitatively measured even in low-MI contrast ultrasonography.


[Private nurses in associations: an enrichment for practice].
After a year of informal meetings between private nurses in the town of Bron (69), the Bronidels association was created. Projects have been developed and completed within this framework, and this structuring has enabled the Idels to be even more reactive in the context of the Covid-19 crisis. Although private practice is above all an isolated practice, the association has highlighted the desire of nursing professionals to come together and the benefits they withdraw from it.


Pregnancy outcomes of multiple repeated cesarean sections in King Chulalongkorn Memorial Hospital.
To determine the incidence of maternal and neonatal morbidity and mortality in women with history of at least one previous cesarean section and to compare the pregnancy outcome of the women with the history of one previous cesarean section to the women with the history of two or more previous cesarean section. We performed a retrospective study of 458 patients undergoing repeated cesarean section from 1998 to 2005. Various factors that may be associated with repeated cesarean sections and pregnancy outcomes were assessed and analyzed. Maternal morbidity rate was 18.6%, including operative complications (17.5%) and post operative complications (1.7%). The operative complications included intraperitoneal adhesion 45 cases (9.8%), excessive blood loss (> 1000 ml) 29 cases (6.3%), placenta previa 5 cases (1.1%), placenta adherens 6 cases (1.3%) and requirement of blood transfusion 10 cases (2.2%). Postoperative complications included postpartum hemorrhage 4 cases (0.9%). No maternal mortality was found in this study. Neonatal morbidity rate was 20.5%, including neonatal jaundice 33 case (7.20), large for gestational age 37 cases (8.1%), and preterm babies 20 cases (4.4%). There were no statistically significant differences of maternal morbidity and neonatal morbidity between the pregnant women with the history of one previous cesarean section and the women with the history of two or more previous cesarean section. Maternal and neonatal morbidity in repeated cesarean section were low.


The impact of sports participation on mental health and social outcomes in adults: a systematic review and the 'Mental Health through Sport' conceptual model.
Sport is a subset of physical activity that can be particularly beneficial for short-and-long-term physical and mental health, and social outcomes in adults. This study presents the results of an updated systematic review of the mental health and social outcomes of community and elite-level sport participation for adults. The findings have informed the development of the 'Mental Health through Sport' conceptual model for adults. Nine electronic databases were searched, with studies published between 2012 and March 2020 screened for inclusion. Eligible qualitative and quantitative studies reported on the relationship between sport participation and mental health and/or social outcomes in adult populations. Risk of bias (ROB) was determined using the Quality Assessment Tool (quantitative studies) or Critical Appraisal Skills Programme (qualitative studies). The search strategy located 8528 articles, of which, 29 involving adults 18-84 years were included for analysis. Data was extracted for demographics, methodology, and study outcomes, and results presented according to study design. The evidence indicates that participation in sport (community and elite) is related to better mental health, including improved psychological well-being (for example, higher self-esteem and life satisfaction) and lower psychological ill-being (for example, reduced levels of depression, anxiety, and stress), and improved social outcomes (for example, improved self-control, pro-social behavior, interpersonal communication, and fostering a sense of belonging). Overall, adults participating in team sport had more favorable health outcomes than those participating in individual sport, and those participating in sports more often generally report the greatest benefits; however, some evidence suggests that adults in elite sport may experience higher levels of psychological distress. Low ROB was observed for qualitative studies, but quantitative studies demonstrated inconsistencies in methodological quality. The findings of this review confirm that participation in sport of any form (team or individual) is beneficial for improving mental health and social outcomes amongst adults. Team sports, however, may provide more potent and additional benefits for mental and social outcomes across adulthood. This review also provides preliminary evidence for the Mental Health through Sport model, though further experimental and longitudinal evidence is needed to establish the mechanisms responsible for sports effect on mental health and moderators of intervention effects. Additional qualitative work is also required to gain a better understanding of the relationship between specific elements of the sporting environment and mental health and social outcomes in adult participants.


Killing activity of human umbilical cord blood-derived TCRValpha24(+) NKT cells against normal and malignant hematological cells in vitro: a comparative study with NK cells or OKT3 activated T lymphocytes or with adult peripheral blood NKT cells.
We aimed to determine the effects of human umbilical cord blood (UCB)-derived natural killer T (NKT) cells as immunological effectors against hematological malignancies, as well as auto- or allo-dendritic cells (DCs) or EB transformed cell lines (EBCLs). TCRValpha24(+) Vbeta11(+) UCB- or PB-NKT cells were isolated by sorting and activated by alpha-galactosylceramide-pulsed autologous DCs. UCB-NK cells were induced from CD34(+) cells by stem cell factor plus IL-15. UCB-T cells were primarily activated by anti-CD3 monoclonal antibody. All those effectors were cultured with IL-2 (100 U/ml), and their cytotoxic activities were evaluated by (51)Cr-release assay. UCB-NKT cells were cultured with IL-12, IL-18 or higher dose of IL-2 (1000 U/ml), and again tested for the cytotoxicity against selected targets. UCB-NKT cells exhibited a pattern of killing activity against various hematological malignancies similar to that of UCB-NK cells, but could not kill K562, which was a vulnerable target for NK cells. The level of activity was quite similar to that of PB-NKT cells. In contrast, OKT-3-activated UCB-T lymphocytes showed a stronger and wider spectrum of killing compared with UCB-NK or NKT cells. IL-12, IL-18 or a higher dose of IL-2 upregulated the activity; however several targets, including fresh leukemic cells, still remained resistant. NKT cells killed auto- or allo-DCs at a level similar to that of T cells, but could not kill allo-EBCLs, which were efficiently killed by T cells. While NK cells showed only marginal or no killing against DC or EBCLs. The anti-cancer activity of human NKT cells depends on the concentrations or the combination of Th1-cytokines. Basically, those cells might not be contributing to the immune surveillance of hematological malignancies, as shown by a relatively low cytotoxicity against malignant cells, together with the quite strong killing against auto-DCs.


sumSTAAR: A flexible framework for gene-based association studies using GWAS summary statistics.
Gene-based association analysis is an effective gene-mapping tool. Many gene-based methods have been proposed recently. However, their power depends on the underlying genetic architecture, which is rarely known in complex traits, and so it is likely that a combination of such methods could serve as a universal approach. Several frameworks combining different gene-based methods have been developed. However, they all imply a fixed set of methods, weights and functional annotations. Moreover, most of them use individual phenotypes and genotypes as input data. Here, we introduce sumSTAAR, a framework for gene-based association analysis using summary statistics obtained from genome-wide association studies (GWAS). It is an extended and modified version of STAAR framework proposed by Li and colleagues in 2020. The sumSTAAR framework offers a wider range of gene-based methods to combine. It allows the user to arbitrarily define a set of these methods, weighting functions and probabilities of genetic variants being causal. The methods used in the framework were adapted to analyse genes with large number of SNPs to decrease the running time. The framework includes the polygene pruning procedure to guard against the influence of the strong GWAS signals outside the gene. We also present new improved matrices of correlations between the genotypes of variants within genes. These matrices estimated on a sample of 265,000 individuals are a state-of-the-art replacement of widely used matrices based on the 1000 Genomes Project data.


Glycated Hemoglobin A1c, Lung Function, and Hospitalizations Among Adults with Asthma.
Insulin resistance and metabolic dysfunction have been associated with asthma risk and asthma severity. To examine the association between glycated hemoglobin A1c (HbA1c), asthma-related hospitalizations, and lung function measures among adults in the United Kingdom. A cross-sectional study was conducted of 47,606 adults aged 40 to 69 years who participated in the UK Biobank and had asthma but no diagnosis of diabetes mellitus. HbA1c level was analyzed as a continuous measure and also categorized as normal (<42 mmol/mol) or as consistent with prediabetes/diabetes (≥42 mmol/mol). An asthma-related hospitalization was defined as ever having had a hospitalization with an International Classification of Diseases, Ninth/Tenth Revision, Clinical Modification code of a main diagnosis compatible with asthma (International Classification of Diseases, Ninth Revision, Clinical Modification code 493.x or International Classification of Diseases, Tenth Revision, Clinical Modification codes J45.x and J46.x). Logistic or linear regression was used for the multivariable analysis of asthma hospitalizations and lung function measures (FEV1, forced vital capacity [FVC], and FEV1/FVC). All models were adjusted for age, sex, ethnic background, body mass index, average annual household income, current smoking status, pack-years of smoking, fasting time, and C-reactive protein level. Both HbA1c level (odds ratio, 1.03; 95% CI, 1.01-1.04) and an HbA1c level in the prediabetes/diabetes range (odds ratio, 1.68; 95% CI, 1.18-2.41) were associated with 1 or more asthma hospitalizations. Moreover, both HbA1c level and an HbA1c level in the prediabetes/diabetes range were significantly and inversely associated with FEV1 and FVC. HbA1c is linked to asthma-related hospitalizations and small decrements in FEV1 and FVC among British adults with asthma but no diagnosis of diabetes mellitus.


Von Willebrand's disease. Clinical picture, diagnosis, and treatment.
Recent developments regarding von Willebrand's disease, a complex disorder of hemostasis, have been helpful in understanding the clinical and laboratory manifestations of the disorder. This article discusses the pathogenesis and classification of the disorder, the role of von Willebrand factor in hemostasis, genetic counseling, and approaches to treatment.


Using Yoga to Reduce Anxiety in Children: Exploring School-Based Yoga Among Rural Third- and Fourth-Grade Students.
Anxiety is becoming more common in children and can negatively affect social and academic performance. The purpose of this study was to explore the impact of 10 min of yoga on anxiety in third- and fourth-graders over 8 weeks. Participants completed 10 min of yoga daily during the school week. Participants completed the Screen for Child Anxiety Related Emotional Disorders anxiety screening tool at the beginning and after the program. Paired sample t test noted a significant difference between pretest and posttest raw scores in the category of generalized anxiety disorder. On average, participants had significantly decreased raw anxiety scores after completing the program (mean = -0.0308), t(60) = -3.137, p < .05. This study demonstrates that yoga practiced as little as 10 min a day over 8 weeks can have a significant impact on decreasing anxiety in children aged 8-10 years.


Total Synthesis of Lycopalhine A.
The total synthesis of lycopalhine A has been accomplished. The synthesis features construction of the tricyclic system via cleavage of a cyclopropane ring and an ensuing intramolecular Michael addition, stereoselective introduction of a 2-aminoethyl moiety via a reaction of allyltrimethylsilane to a sulfonyliminium ion, and a stereoselective intramolecular aldol reaction.


Where is the patient in diabetes performance measures? The case for including patient-centered and self-management measures.
Health policies are important determinants of clinician and patient behavior, and an important policy issue is what items are included in healthcare quality and performance measures. There is consensus that patient-centered care and self-management support are essential evidence-based components of good diabetes care. However, most major diabetes performance measures such as the National Committee for Quality Assurance (NCQA)/American Diabetes Association (ADA) Provider Recognition Program indexes have not included self-management or psychosocial items. We review the case for and propose a set of patient-centered, self-management indicators to be included as a standard part of diabetes quality indicators. The proposed indicators include: patient self-management goal(s), measures of health behaviors (e.g., healthy eating, medication taking, physical activity, and smoking status), quality of life, and patient-centered collaborative care. We discuss the evidence and the concerns about patient-report measures and summarize successful incorporation of such patient-centered measures in other countries and by the American Association of Diabetes Educators (AADE). The adage that "what gets measured, gets done" applies to diabetes management and many other areas of healthcare. Inclusion of the proposed indicators in national diabetes performance measures would be consistent with Institute of Medicine (IOM), ADA, Centers for Disease Control (CDC), Diabetes Attitudes, Wishes, and Needs (DAWN), AADE, and Society of Behavioral Medicine (SBM) recommendations. Such action would enhance both the priority and delivery of quality, patient-centered care, and diabetes self-management support.


Exploring factors underlying the attitude of community pharmacists to generic substitution: a nationwide study from Poland.
Generic uptake will increasingly be promoted by governments in the face of increasing healthcare costs and global economic uncertainties. The purpose of this study was to investigate attitudes towards generic substitution among community pharmacists, with a focus on the perception of the efficacy, knowledge of the generics characteristics, as well as the willingness to recommend generic substitution. Community pharmacies in Poland. The survey was conducted in 2013 by telephone interviews with 802 holders of an MSc degree in pharmacy working as community pharmacists. Stratified sampling was implemented to make the study representative in geographic terms. Pharmacists' attitudes towards generics drugs. The study showed that only 40 % of pharmacists always inform patients about their right to choose a generic substitute. It was also shown that the less time a pharmacist has been practising, the less likely they are to invite consumers to choose between generic and innovator products. The likelihood of informing was not affected by pharmacist's sex or age, or by pharmacy location or status (chain vs. independent pharmacy) (p > 0.05). Pharmacists varied in their approach to their statutory obligation to inform about a generic; a more or less equal share of respondents were either in favour or against it. Approximately 60 % pharmacists were shown to be familiar with the definition of a generic medicine. Pharmacists with shorter time of practice proved to know more about generics. However, more than 30 % respondents failed to choose the correct statement on generic versus reference medicine dosage. The majority of respondents (67 %) believed there are no differences in efficacy between generics and innovator drugs, whereas 31 % claimed that original brands could be more effective. A significant correlation was demonstrated between the views of pharmacists on the therapeutic efficacy and their willingness to substitute for generics whenever permitted by a physician. It is important to address all concerns pharmacists may have over generics, for example by implementing comprehensive awareness-raising campaigns. Also, pharmacotherapy monitoring systems (i.e. provided in a framework of pharmaceutical care) could be considered to identify any safety or quality concerns that may arise.


Osteoporosis unmasked by hyperthyroidism in a young man with osteogenesis imperfecta.
A young man presented with a vertebral fracture and generalized osteopenia that was initially attributed to coexisting hyperthyroidism. However, the rarity of fractures in young men with hyperthyroidism and the finding of blue sclerae led to the diagnosis of osteogenesis imperfecta. Since vertebral fractures are uncommon in young men with osteogenesis imperfecta, we postulate that the combined effects of the two disorders contributed to his clinical presentation.


Motor functioning during and following treatment with chemotherapy for pediatric acute lymphoblastic leukemia.
This systematic review evaluated empirical studies examining motor skills in children during and following treatment for acute lymphoblastic leukemia. Most studies indicated that children on-treatment display poorer gross and fine motor abilities than healthy peers, but generally have intact visual-motor integration skills. Studies have reported gross motor difficulties in 5-54% of survivors. There is some limited evidence for long-term fine motor deficits. The evidence for visual-motor integration difficulties in the survivor population is less consistent. Larger studies with a longitudinal design are needed to further specify the onset and timing of motor difficulties and ascertain risk factors.


Donor T cells administered over HLA class II barriers mediate antitumor immunity without broad off-target toxicity in a NOD/scid mouse model of acute leukemia.
Alloreactive (allo)-HLA-directed T cell responses after HLA-mismatched allogeneic hematopoietic stem cell transplantation and donor lymphocyte infusion are typically considered detrimental responses mediating graft-versus-host disease (GVHD). Allo-HLA-reactive T cells with beneficial and selective graft-versus-leukemia (GVL) reactivity, however, can also be identified within an HLA-mismatched context. We investigated whether allo-HLA class II-directed T cells with beneficial GVL reactivity induced in NOD/scid mice engrafted with human chronic myelogenous leukemia in lymphoid blast crisis after treatment with donor lymphocyte infusion - mediated detrimental xenogeneic GVHD as a result of broad off-target cross-reactivity. The results demonstrate that beneficial GVL reactivity and xenogeneic GVHD are mediated by separate T cells. GVL reactivity was mediated by human T cells recognizing allo-HLA class II molecules, whereas xenoreactivity was exerted by human T cells recognizing H-2 molecules. Taken together, our data indicate a limited risk for detrimental off-target effects by allo-HLA class II-directed T cells and thereby provide a basis for the development of strategies for selecting allo-HLA-restricted T cells with selective GVL reactivity for adoptive transfer after HLA-mismatched allogeneic hematopoietic stem cell transplantation.


Visual localization of briefly presented peripheral targets.
Reported here are the results from two experiments designed to investigate Mateeff and Gourevich's (1983, 1984) claim that adult observers make large constant errors when judging the direction of briefly presented peripheral targets, with respect to a continuously visible scale. Experiment 1 involved a virtually exact duplication of Mateeff and Gourevich's paradigm. Experiment 2 involved adjusting the position of a light emitting diode (LED) so as to match the direction of a LED previously flashed in the periphery. No significant constant errors were found in either experiment.


Minimally invasive oesophagectomy: The first case report of a thoracolaparoscopic oesophagectomy done in the Caribbean.
Minimally invasive oesophagectomy is one of the options in the armamentarium of a modern surgeon when faced with treating oesophageal cancer. Trinidad and Tobago has successfully treated a patient using this method. This case report adds to the rich surgical history in the Caribbean. Our patient presented with progressive dysphagia, and after being worked up was diagnosed with oesophageal cancer. He underwent neoadjuvant chemotherapy and had a minimally invasive McKeown oesophagectomy done by a fellowship trained, advanced laparoscopic oesophageal and gastric surgeon. The patient had an uneventful recovery and was tumor free with excellent quality of life at 4 years, showing that the Caribbean is keeping up with the rest of the surgical world.


The stimulation of pp60v-src kinase activity by vanadate in intact cells accompanies a new phosphorylation state of the enzyme.
We have observed that intact wild type Rous sarcoma virus-transformed chicken embryo fibroblasts (CEF), when incubated with micromolar sodium orthovanadate for as little as 4 h, results in a 2-3-fold increase in pp60v-src kinase activity as judged by the IgG kinase assay. The addition of sodium vanadate or vanadyl sulfate to transformed cell lysates or the immunoprecipitate assay was without effect on the activity of isolated pp60v-src kinase activity. The increased kinase activity is reflected intracellularly by an increased phosphotyrosine content of a known substrate of pp60v-src, a 36-kDa phosphoprotein, without a change in total phosphate incorporation into cellular proteins. Pulse and pulse-chase experiments with [35S]methionine indicated that there was no change in the net or rate of biosynthesis of pp60v-src which might have accounted for the elevated kinase activity. Along with this stimulation of pp60v-src kinase activity, we found a parallel increase in the phosphate content of the enzyme and the appearance of an electrophoretic variant after vanadate treatment. The increased phosphorylation of the kinase is accounted for by an increase in phosphotyrosine content without a change in phosphoserine content. The increased phosphotyrosine content is shown to be resident in the carboxyl-terminal fragment and, unexpectedly, in the amino-terminal fragment of the kinase. The phosphotyrosine present in the amino-terminal portion may be a de novo phosphorylation, but could also represent a tyrosine site(s) not easily detected under unstimulated conditions in the absence of vanadate.


Efficacy and safety of sacubitril/valsartan compared with enalapril in patients with chronic heart failure and reduced ejection fraction: Results from PARADIGM-HF India sub-study.
To determine efficacy and safety of sacubitril/valsartan compared with enalapril in Indian patients of PARADIGM-HF trial. A randomized, double-blind, active-controlled, phase III sub-study (NCT01035255) was conducted between April 2010 and May 2014. Patients with chronic heart failure (HF), aged >18 years with left ventricular ejection fraction ≤40% were randomized (1:1) to receive either sacubitril/valsartan 200 mg twice-daily or enalapril 10 mg twice-daily. The primary endpoint was to compare efficacy of sacubitril/valsartan to enalapril in delaying time-to-first occurrence of the composite endpoint (cardiovascular [CV] death or HF hospitalization). The trial was stopped after a median follow-up of 27 months, because the boundary for benefit with sacubitril/valsartan had crossed. Among 637 Indian patients in PARADIGM-HF (sacubitril/valsartan, n = 322 and enalapril, n = 315), the primary outcome, CV death, and the first hospitalization for HF occurred in 21.81% and 24.76% (HR 0.89; 95% CI, 0.646-1.231), 17.45% and 20.63% (HR 0.87; 95% CI, 0.605-1.236), and 7.48% and 9.52% (HR 0.78; 95% CI, 0.461-1.350) patients in the sacubitril/valsartan and enalapril group, respectively. The all-cause mortality (19.0% vs. 21.9%) and adverse events (78.4% vs. 82.2%) were comparatively lower in the sacubitril/valsartan than enalapril group. No significant difference was seen between the benefits of treatment in Indian and the total PARADIGM-HF cohort (p value for interaction >0.05). Results support the use of sacubitril/valsartan in Indian patients with chronic HF with reduced ejection fraction with treatment benefits similar to global PARADIGM-HF cohort.


Neurophilia is stronger for educators than students in Brazil.
Educational products claiming to be "brain-based" are common. Due to neurophilia, including a brain in a product's marketing can enhance perceptions. However, schooling background may play a protective role. As previous neuromarketing research has been conducted predominantly in English speakers, we examined whether the effects of neurophilia extend to a Portuguese-speaking Brazilian population. Teachers and students (N = 262) viewed one of four advertisements for a hypothetical product translating to ''Right Brain'' or ''Right Start'' Training; half the advertisements contained an MRI brain image. Participants rated their perceptions of interest, efficacy, and scientific rationale. The presence of a brain image or the word 'brain' did not influence responses. However, occupation had a significant effect: teachers' ratings were higher than students' ratings. Importantly, teachers were more susceptible to neurocontent, rating "Right Brain" training significantly higher than students. These results thus highlight the need to improve teachers' neuroscience literacy.


[Neurostimulation of the recurrent laryngeal nerve--a routine method in thyroid gland surgery?].
Thyroid operations belong to the most frequent procedures in Germany (100,000/year). An important quality parameter is the incidence of postoperative recurrent nerve paralysis. Intraoperative identification by visualization of the nerve with a paralysis rate of 1% is the present gold standard in dedicated centers. Can this results be further improved by use of an intraoperative neuromonitoring system (NM). The use of NM is helpful in difficult situations (recurrent goiter, advanced carcinoma, anatomic variants), but a quality improvement is not yet proven. NM can not replace the current nerve identification by meticulous preparation of anatomic structures. It should be used at the discretion of the operative surgeon.


The effect of growth hormone on sleep-related cardio-respiratory control in Prader-Willi syndrome.
To evaluate the effects of growth hormone (GH) treatment on control of breathing, heart rate and blood pressure during sleep in Prader-Willi Syndrome (PWS). In a prospective clinical case series study, sixteen consecutive PWS patients (median age 16 months at enrolment) were followed-up 6 months (2-32 months) after commencing GH treatment. We compared heart rate (HR), Pulse Transit Time (PTT; an index of blood pressure, BP) and ventilatory responses to standard chemostimuli (4% CO(2) and 100% O(2)) during quiet sleep prior to and after commencing GH treatment. Growth hormone treatment increased arterial oxygenation during sleep but did not significantly improve breathing stability (apnoea-hypopnoea index remained unchanged). GH treatment did not alter ventilatory, HR and PTT chemoreceptor-mediated responsiveness (p = 0.23-0.97) but did significantly improve the coupling between and HR and PTT, indicating that HR and BP rose (or fell) in parallel after but not before GH therapy (p = 0.01). Growth hormone treatment improves arterial oxygenation and cardiovascular function during sleep; these changes are not owing to improved (stronger) chemoreflex-mediated autonomic drive.


Reducing the barriers to RN educational mobility.
How can the needs of RN students be met without compromising the quality of the educational experience? The model of flexible clinical scheduling described here has built-in safeguards to preserve quality and promote flexibility. The author discusses the benefits for both students and faculty.


A robust optimization model for microgrid considering hybrid renewable energy sources under uncertainties.
Hybrid renewable energy sources and microgrids will determine future electricity generation and supply. Therefore, evaluating the uncertain intermittent output power is essential to building long-term sustainable and reliable microgrid operations to fulfill the growing energy demands. To address this, we proposed a robust mixed-integer linear programming model for the microgrid to minimize the day-ahead cost. To validate the proposed model piecewise linear curve is to deal with uncertainties of wind turbine, photovoltaic, and electrical load. The proposed solution is demonstrated through a case study compared under a robust worst-case scenario, deterministic model, and max-min robust optimization that aim to find optimal robustness. So, a piecewise linear curve is considered to obtain uncertain parameters in order to deal with uncertainties and predict the day-ahead cost. This study illustrates how the Uncertainty Budget Set selection used to integrate renewable energy sources into a microgrid, which manages the energy system. Therefore, the model complexity was slightly modified by adjusting the Uncertainty Budget Set to obtain the optimal decision and control the load demand and uncertainty of renewable energy sources. The comparative results demonstrate that the proposed robust optimization can achieve high solutions under microgrid's availability and is intended to confirm that the proposed method is more cost-effective than alternative optimization techniques. Additionally, the effectiveness and advantage of the proposed methodology in the IEEE 33-node system are validated in this case study by comparing it to the existing optimization. The comparison results show that the proposed robust optimization methods illustrate the model's efficiency, concluding remarks, and managerial insights of the research.


Functional analysis of an overexpressed glutathione S-transferase BdGSTd5 involved in malathion and malaoxon detoxification in Bactrocera dorsalis.
Glutathione S-transferases (GSTs) are one of the three detoxification enzyme families. The constitutive and inducible overexpression of GSTs genes plays an important role in insecticide resistance. Previous study showed that malathion resistance was polygenic, and elevated GSTs activity was one of the important factor participating in malathion resistance of Bactrocera dorsalis (Hendel), a serious economic pest worldwide. BdGSTd5 overexpression was inducible upon exposure to malathion. However, the involvement of BdGSTd5 in malathion resistance has not been clarified. In this study, we found that BdGSTd5 sequence harbored the conserved region of delta class GSTs, which were overexpressed in malathion resistant strain of B. dorsalis compared to malathion susceptible strain. The highest mRNA expression level of BdGSTd5 was found in 1-day-old adult, and the levels decreased with aging. The dsBdGSTd5 injection effectively silenced (73.4% reduction) the expression of BdGSTd5 and caused significant increase in susceptibility to malathion with a cumulative mortality increasing of 13.5% at 72 h post malathion treatment (p < 0.05). Cytotoxicity assay demonstrated that BdGSTd5 was capable of malathion detoxification. Molecular docking analysis further indicated the interactive potential of BdGSTd5 with malathion and its toxic oxide malaoxon. The recombinant BdGSTd5 exhibited glutathione-conjugating activity toward 1-chloro-2, 4-dinitrobenzene and malathion and malaoxon metabolic capacity with significant reduction (p < 0.05) of the peak areas by 90.0% and 73.1%, respectively. Taken together, the overexpressed BdGSTd5 contributes to malathion metabolism and resistance, which detoxify the malathion in B. dorsalis via directly depleting malathion and malaoxon.


Durability of every-8-week aflibercept maintenance therapy in treatment-experienced neovascular age-related macular degeneration.
To determine the proportion of treatment-experienced eyes with exudative age-related macular degeneration successfully treated with every-4-week aflibercept that can be kept dry on fixed every-8-week aflibercept injections (maintenance). In this retrospective chart review, we evaluated our cohort of patients treated with a treatment paradigm for CNV in AMD. Initially, patients were treated with bevacizumab or ranibizumab and switched to every-4-week aflibercept when therapeutic responses were not durable or were suboptimal. Maintenance every-8-week therapy was initiated when the retina was completely dry on every-4-week aflibercept therapy. The primary outcome measure was recurrence of exudation on optical coherence tomography (OCT) during maintenance. Thirty-six eyes of 31 consecutive patients with median age of 79 years (range, 65-89) were included. Maintenance was started after a median of 34 (range, 8-88) injections. Recurrence was observed in 20 eyes (55%). Of these, 11 eyes (31%) reactivated at 8 weeks. Median time to failure of maintenance schedule was 40 weeks by Kaplan-Meier analysis. Baseline demographic and anatomic characteristics were not associated with failure of maintenance schedule. In treatment-experienced eyes that respond completely to every-4-week aflibercept, maintenance therapy with every-8-week injections can only temporarily maintain anatomic success with the majority of eyes developing recurring activity. This regimen fails early in one third of eyes and has a median effective duration of 40 weeks. Aflibercept appears to be inadequate to maintain control of exudation in most eyes in at least half of eyes undergoing long-term therapy.


Catalyzing community action within a national campaign: VERB community and national partnerships.
The VERB campaign used a social marketing approach to deliver its message through the mass media, school and community promotions, and partnerships to encourage children aged 9-13 years (tweens) to be physically active every day. This paper presents the VERB campaign's community and national partnership strategy, highlights three successful partnerships, and discusses challenges associated with the efforts. The national advertising generated awareness of and affinity for the product's brand and motivated the primary audience to seek out the product. The campaign's national and community partners were engaged to facilitate a product-distribution channel. The campaign developed a three-pronged partnership strategy to integrate the promotion with the placement of the campaign's product (physical activity): (1) reframe the way physical activity is positioned and delivered; (2) connect the brand to the point-of-purchase; and (3) refer (or drive) the audience to the action outlets, opportunities, places, spaces and programs to purchase the product. The VERB campaign provided partners with marketing training and resources to assist them as they leveraged tweens' brand awareness and supported regular physical activity among tweens. The method of technical assistance and the types of marketing tools were provided in relationship to four characteristics of the partner: (1) partner's network, (2) leaders and champions in the network, (3) partner's financial resources for community campaigns; and (4) partner's understanding of the marketing mindset. Coordinated, collaborative, and strong mass-media and community-based interventions within a national social marketing campaign can sustain the immediate effects of such campaigns.


Assessment of heat and storage conditions on gamma-ray and electron beam irradiated UHMWPE by electron spin resonance.
Electron spin resonance (ESR) spectroscopic study was undertaken to explore the nature of any remaining radicals in ultra-high molecular weight polyethylene (UHMWPE) after irradiation with gamma-rays and electron beams to a dose of 25 KGy in air or N2 environment. The decay of radicals was studied by observing the ESR intensity change as a function of time. The following post-irradiation conditions were employed: (1) storage in air or N2 at 25 degrees C, and (2) heat treatment in air or N2 at 80, 100, and 120 degrees C for time intervals up to 8 h. The study suggests that radicals remaining trapped in the matrix of UHMWPE could be dramatically scavenged by heat treatment, independently of the atmosphere during irradiation and the heating process. The melting temperature and mechanical properties of irradiated UHMWPE under the experimental conditions employed were shown not to alter significantly by heat treatment, except in the presence of air.


Additive lymph node dissection may be necessary in minute submucosal cancer of the stomach after endoscopic resection.
In early gastric cancer (EGC), minute submucosal (SM1) invasion of the stomach has been regarded as an expanded indication for endoscopic resection (ER). The exact prediction of SM1 invasion before ER may be difficult. Thus, SM1 invasion may be important to decide additive treatment after ER. This study was designed to investigate the incidence of lymph node metastasis (LNM) in SM1-EGC based on surgical specimens and to evaluate the factors that indicate additional treatment after ER. From May 2005 to December 2008, 1,676 patients with EGC underwent surgery at Severance and Gangnam Severance Hospital, Seoul, Korea. Among them, 126 patients were diagnosed with differentiated SM1-EGC. The clinicopathologic characteristics were analyzed with respect to LNM and lymphovascular invasion (LVI), which is a known independent risk factor for LNM. Intratumoral marker immunohistochemistry was examined as a predictor of LVI. The mean SM1 invasion depth was 621.3 ± 745.6 μm. The LNM rates did not differ significantly between differentiated SM1-EGC (6.3%) and SM1-EGC (4.1%) meeting the expanded indication for ER. Female gender, moderate differentiation, LVI, and LVI grade were positively correlated with LNM. Female gender and elevated lesion morphology were associated with LVI. The expression levels of VEGF-C and OPHN1 were higher in LVI-positive tissues. The LNM rate in differentiated SM1-EGC meeting the expanded ER criteria was 4.1% in the present study, indicating that additional lymph node dissection may be necessary after ER in some cases of SM1-EGC.


Properties of (Na+ plus K+)-activated ATPase in rat liver plasma membranes enriched with bile canaliculi.
Liver plasma membranes enriched in bile canaliculi were isolated from rat liver by a modification of the technique of Song et al. (J. Cell Biol. (1969) 41, 124-132) in order to study the possible role of ATPase in bile secretion. Optimum conditions for assaying (Na+ plus K+)-activated ATPase in this membrane fraction were defined using male rats averaging 220 g in weight. (Na+ plus K+)-activated ATPase activity was documented by demonstrating specific cation requirements for Na+ and K+, while the divalent cation, Ca(2+), and the cardiac glycosides, ouabain and scillaren, were inhibitory. (Na+ plus K+)-activated ATPase activity averaged 10.07 plus or minus 2.80 mumol Pi/mg protein per h compared to 50.03 plus or minus 11.41 for Mg(2+)-activated ATPase and 58.66 plus or minus 10.07 for 5'-nucleotidase. Concentrations of ouabain and scillaren which previously inhibited canalicular bile secretion in the isolated perfused rat liver produced complete inhibition of (Na+ plus K+)-activated ATPase without any effect on Mg(2+)-activated ATPase. Both (Na+ plus K+)-activated ATPase and Mg(2+)-activated ATPase demonstrated temperature dependence but differed in temperature optima. Temperature induced changes in specific activity of (Na+ plus K+)-activated ATPase directly paralleled previously demonstrated temperature optima for bile secretion. These studies indicate that (Na+ plus K+)-activated ATPase is present in fractions of rat liver plasma membranes that are highly enriched in bile canaliculi and provide a model for further study of the effects of various physiological and chemical modifiers of bile secretion and cholestasis.


Leptin-induced signal transduction pathways.
Leptin is a multifunctional cytokine and hormone that primarily acts in the hypothalamus and plays a key role in the regulation of food intake and energy expenditure. In addition, it has direct effects on many cell types on the periphery. Leptin acts through its receptor, the product of the db gene, which has six isoforms. Only one of them (OB-Rb) has full signalling capabilities and is able to activate the Jak/STAT pathway, the major pathway used by leptin to exert its effects. However, some signalling events can be initiated by the short isoforms. Besides Jak/STAT, other pathways, such as MAPK and the 5'-AMP-activated protein kinase (AMPK) pathway, are also involved in leptin signalling. Leptin also interacts with insulin signalling. In this paper, we give an overview of the signal transduction mechanisms that are related to the actions of leptin.


Genetic manipulation of ureteric bud tip progenitors in the mammalian kidney through an Adamts18 enhancer driven tet-on inducible system.
The ureteric epithelial progenitor (UEP) population within the embryonic kidney generates the arborized epithelial network of the kidney's collecting system and plays a critical role in the expansion and induction of the surrounding nephron progenitor pool. Adamts18 shows UEP- restricted expression in the kidney and progenitor tip-restricted expression in several other organs undergoing branching epithelial growth. Adamts18 is encoded by 23 exons. Genetic removal of genomic sequence spanning exons 1 to 3 led to a specific loss of Adamts18 expression in UEPs, suggesting this region may encode a UEP-specific enhancer. Intron 2 (3 kb) was shown to have enhancer activity driving expression of the doxycycline inducible tet-on transcriptional regulator (rtTA) in an Adamts18en-rtTA transgenic mouse strain. Crossing Adamts18en-rtTA mice to a doxycycline dependent GFP reporter mouse enabled the live imaging of embryonic kidney explants. This facilitated the analysis of ureteric epithelial branching events at the cellular level. Ablation of UEPs at the initiation of ureteric bud outgrowth through the doxycycline-mediated induction of Diphtheria Toxin A (DTA) generated a range of phenotypes from complete kidneys agenesis, to duplex kidneys with double ureters. The latter outcome points to the potential of regulative processes to restore UEPs. In contrast, overexpression of YAP prior to ureteric bud outgrowth led to a complete failure of kidney development. Elevating YAP levels at later stages retarded branching growth. A similar phenotype was observed with the overexpression of MYC within the branch-tip localized UEP population. These experiments showcase the utility of the Adamts18en-rtTA transgenic model to the investigation of cellular and molecular events specific to branch tip progenitors within the mammalian kidney complementing existing CRE-dependent genetic tools. Further, the illustrative examples point to areas where new insight may be gained into the regulation of UEP programs.


Total antioxidant status and oxidative stress in recurrent aphthous stomatitis.
Recurrent aphthous stomatitis is an idiopathic, chronic, recurrent inflammatory disease of the oral mucosa. It is thought that oxidative stress caused by systemic inflammation plays a basic role in the etiopathogenesis of recurrent aphthous stomatitis. The aim of this study is to review oxidative status and DNA damage in recurrent aphthous stomatitis. The study included 42 patients with an active recurrent aphthous stomatitis lesion and 39 healthy volunteers with similar demographic characteristics. DNA damage was analyzed using alkaline single cell gel electrophoresis (comet assay). Plasma levels of total antioxidant status and total oxidative status were determined by using an automated measurement method. Oxidative stress index was calculated as total oxidative status/total antioxidant status and × 100. The total oxidative status and oxidative stress index values were significantly higher in the recurrent aphthous stomatitis group compared to the control group, while total antioxidant status values were significantly lower. In the recurrent aphthous stomatitis group, DNA damage was observed to be significantly higher than the control group. In correlation analysis, significant correlation was found between DNA damage and the oxidative stress index and total oxidative status values in the recurrent aphthous stomatitis group. This is the first report in the literature that demonstrates association of recurrent aphthous stomatitis with increased oxidative status.


Self-efficacy, support and sustainability - a qualitative study of the experience of establishing breastfeeding for first-time Australian mothers following early discharge.
Ensuring women receive optimal breastfeeding support is of key importance to the health of mothers and their infants. Early discharge within 24 h of birth is increasingly common across Australia, and the practice of postnatal home visiting varies between settings. The reduction in length of stay without expansion of home visits reduces midwives' ability to support breastfeeding. The impact of early discharge on first-time mothers establishing breastfeeding was unknown. The study aim was to understand the experiences of first-time Australian mothers establishing breastfeeding when discharged from the hospital within 24 h of a normal vaginal birth. A qualitative interpretive method was used. Semi-structured interviews with 12 women following early discharge were conducted. Data were audio recorded, professionally transcribed, and subjected to a thematic analysis. Three interconnected themes of 'self-efficacy', 'support' and 'sustainability' were identified. Self-efficacy influenced the women's readiness and motivation to be discharged home early and played a role in how some of the mothers overcame breastfeeding challenges. Social, semi-professional and professional breastfeeding supports were key in women's experiences. Sustainability referred to and describes what women valued in relation to continuation of their breastfeeding journey. This study found accessible people-based breastfeeding services in the community are valued following early discharge. Furthermore, there is demand for more evidence-based breastfeeding educational resources, potentially in the form of interactive applications or websites. Additionally, a focus on holistic and individualised breastfeeding assessment and care plans prior to discharge that link women with ongoing breastfeeding services is paramount.


Psychological understanding of anthropomorphism in theatre architecture.
Traces of collective projective mechanisms in architecture in general are sedimenting for millenniums. Such case is also with theatre, an old institution of a great social importance. This paper discovers anthropomorphic elements of theatrical architecture. They have been transformed through history into patterns outlined and explained by the author.


Rapid cycling bipolar affective disorder and recurrent strokes secondary to high blood homocysteine.
The interface between psychiatric disorders and organicity has been a matter for contentious debate. To report an interesting clinical case of moderate homocystinuria presenting with significant psychiatric and neurological deficits. A case report highlighting the impact of homocystinuria on producing intractable rapid cycling bipolar affective disorder. Homocystinuria is a frequently missed cause for treatment-resistant bipolar affective disorder.


Characterization of the antifungal and antiprotozoal antibiotic partricin and structural studies on partricins A and B.
Partricin, a heptaene macrolide antibiotic, has been separated into three polyene components, partricins A, B and C, and one non-polyene component by countercurrent distribution. Treatment of partricin with base gave p-(methylamino)acetophenone and p-aminoacetophenone from partricins A and B, respectively, identifying both as members of the aromatic subgroup of the heptaene antibiotics. Both partricins A and B yield mycosamine on mild acid hydrolysis. NMR and mass spectral studies on products of ozonolysis or hydrogenolysis of acetyl derivatives provided evidence for the partial structures 1 approximately 9.


Nuclear envelope permeability measured by fluorescence microphotolysis of single liver cell nuclei.
Fluorescence microphotolysis ("photobleaching") has been widely used to measure translational diffusion coefficients of lipids and proteins in cell membranes. This communication shows that fluorescence microphotolysis can be also employed for measurement of membrane transport in single cells and organelles. The influx of fluorescently labeled dextrans of graded molecular size into leaky human erythrocyte ghosts and isolated rat liver cell nuclei has been measured. For the nuclear envelope, a functional pore radius of 56-59 A is derived.


Bis(nitrato-κO)(1,4,8,11-tetra-aza-cyclo-tetra-decane-κ4 N)zinc(II) methanol monosolvate.
The two ZnII atoms in the crystal structure of the title complex, [Zn(NO3)2(C10H24N4)]·CH3OH, have a distorted octa-hedral coordination sphere, defined by 1,4,8,11-tetra-aza-cyclo-tetra-decane (cyclam) N atoms in the equatorial plane and nitrate O atoms in the axial sites. The conformation of the cyclam is trans-III (R, R, S, S), which is typical for metal-cyclam complexes. Nitrate anions are involved in intra- and inter-molecular hydrogen bonding with the N-H groups of the ZnII-cyclam unit. Together with the methanol solvent mol-ecule, the hydrogen-bonding network connects the ZnII-cyclam units into ribbons running parallel to the a axis.


Structural characterization of the core oligosaccharide isolated from the lipopolysaccharide of the haloalkaliphilic bacterium Salinivibrio sharmensis strain BAG(T).
Salinivibrio genus is included in the family Vibrionaceae and up to now is constituted by only five members. All the species are moderately halophilic bacteria found in salted meats, brines, and several hypersaline environments. Halophilic microorganisms are good sources of biomolecules, such as proteases, that have a great industrial interest as demonstrated by recent studies. All these bacteria possess on their outer membrane amphiphilic molecules named lipopolysaccharides, which are of great interest because of their involvement in the mechanisms of interaction between the microbial life and environmental factors. A novel haloalkaliphilic, facultative anaerobic and Gram-negative Salinivibrio-like microorganism, named S. sharmensis strain BAG(T), was recovered from a saline lake in Ras Mohammed Park (Egypt). The aim of this work is the isolation and structural characterization of the core oligosaccharidic fraction of the lipopolysaccharide from this bacterium. By means of HPAEC-PAD we were able to purify two glycoforms, fully depicted by ESI FT-ICR mass spectrometry, chemical analysis, and NMR spectroscopy. Like other haloalkaliphilic bacteria, the core region was found to be characterized by the presence of several negatively charged residues, such as uronic acids. All the data contributed to give the following structure α-D-Glc-(1-->4)-β-D-GalNAc-(1--4)-β-D-Glc1-->4α-D-GlcA-(1-->2)-α-L,D-Hep-(1-->3)-α-D,D-Hep-(1-->5)-α-D-Kdo4P-(2-->6)-LipidA2<--1β-D-GlcA.


Truncated hemoglobin HbN protects Mycobacterium bovis from nitric oxide.
Mycobacterium tuberculosis, the causative agent of human tuberculosis, and Mycobacterium bovis each express two genes, glbN and glbO, encoding distantly related truncated hemoglobins (trHbs), trHbN and trHbO, respectively. Here we report that disruption of M. bovis bacillus Calmette-Guérin glbN caused a dramatic reduction in the NO-consuming activity of stationary phase cells, and that activity could be restored fully by complementing knockout cells with glbN. Aerobic respiration of knockout cells was inhibited markedly by NO in comparison to that of wild-type cells, indicating a protective function for trHbN. TyrB10, which is highly conserved in trHbs and interacts with the bound oxygen, was found essential for NO consumption. Titration of oxygenated trHbN (trHbN.O(2)) with NO resulted in stoichiometric oxidation of the protein with nitrate as the major product of the reaction. The second-order rate constant for the reaction between trHbN.O(2) and NO at 23 degrees C was 745 microM(-1).s(-1), demonstrating that trHbN detoxifies NO 20-fold more rapidly than myoglobin. These results establish a role for a trHb and demonstrate an NO-metabolizing activity in M. tuberculosis or M. bovis. trHbN thus might play an important role in persistence of mycobacterial infection by virtue of trHbN's ability to detoxify NO.


Size-switchable polymer-based nanomedicines in the advanced therapy of rheumatoid arthritis.
Chronic inflammatory diseases such as rheumatoid arthritis represent a substantial socio-economic impact and have a high prevalence in the modern world. Nano-sized polymer therapeutics have shown suitable characteristics for becoming the next generation of anti-inflammatory nanomedicines. Here, we present biocompatible and stimuli-sensitive N-(2-hydroxypropyl)methacrylamide based polymer conjugates with the anti-inflammatory drug dexamethasone (DEX), which has been tailored for prolonged blood circulation, enhanced inflammatory site accumulation, site-specific drug release and subsequent elimination of the carrier via urine excretion. The hydrodynamic size of novel polymer-DEX nanomedicine was adjusted to prolong its blood circulation whilst maintaining the renal excretability of the polymer carrier after drug release in inflamed tissue. The therapeutic efficacy of the studied polymer nanomedicines was evaluated in a model of dissipated chronic arthritis, i.e. collagen II-induced arthritis, in mice. The pH-sensitive drug attachment enabled enhanced blood circulation with minimal systemic drug release, as well as rapid drug activation in affected joints. Importantly, unlike free DEX, the polymer nanomedicines were able to diminish joint inflammation and arthritis-induced bone damage - even at a reduced dosing regimen - as evaluated by micro computed tomography (micro-CT).


[Sacral stress fracture in a recreational long-distance runner].
The current boom in recreational sports leads to an increase in the number of long-distance runners. In addition to typical disease patterns (e.g. of the Achilles tendon) stress fractures are seen more and more frequently. These mostly occur in the lower leg and foot bones and are rarely seen in the pelvis (e.g. sacrum). Atypical and protracted complaints therefore require a detailed medical history of the patient and should if in any doubt lead to radiological imaging. Once correctly diagnosed, rest from sports and sometimes semi-weightbearing or non-weightbearing is indicated to prevent the development of fracture non-union.


Introduction to microRNAs: Biogenesis, Action, Relevance of Tissue microRNAs in Disease Pathogenesis, Diagnosis and Therapy-The Concept of Circulating microRNAs.
MicroRNAs as the endogenous mediators of RNA interference have principal roles in gene expression regulation. Since their discovery in the early 1990s, their number has steadily grown to approximately 2500 known microRNAs at present in humans. MicroRNAs encoded by distinct genes regulate the expression of about 30-60 % of human protein coding genes by targeting their messenger RNAs (mRNAs) and induce mostly posttranscriptional inhibition, or in some cases enhancement. MicroRNAs, as fine regulators of the gene expression, have important roles in development, the physiological functioning of the organism, e.g. organogenesis, immune functioning, vascular system, etc. The deregulation of microRNA expression has been observed in many disorders, such as in carcinogenesis. Given their tissue specificity and stability, and specific disease-related alterations, tissue microRNAs can be exploited as excellent biomarkers in the diagnosis. Moreover, microRNAs might also be envisaged as novel therapeutic targets. Beside tissue microRNAs, novel data show that microRNAs are also present in body fluids that could further extend their diagnostic utility as minimally invasive biomarkers of various diseases, but also raises intriguing questions regarding their biological relevance. In this introductory chapter, we summarise the most relevant features of microRNAs including their biogenesis, action, the biological, pathological, diagnostic and potential therapeutical relevance of tissue microRNAs.


Prenatal diagnosis of a short-rib-polydactylia syndrome type Saldino-Noonan at 17 weeks' gestation.
A case is described in which the family history and the sonographic findings during the 17th gestational week led to the suspicion of a short-rib-polydactylia syndrome of the Saldino-Noonan type and an abortion was induced. The ultrasound image disclosed cystic structures in the topographic area of the kidney, marked ascites and oligohydramnios and indicated a severe fetal developmental disturbance. Due to the oligohydramnios the extremities could not be interpreted sonographically. The pathological-anatomical findings confirmed the suspected short-rib-polydactylia syndrome of the Saldino-Noonan type. Typical skeletal changes, dysgenetic cystic kidneys and pancreatic fibrosis were especially indicative of the syndrome. The complete agenesia of the islets of Langerhans, present in our observation, has, as far as we know, previously not been reported in the case of short-rib-polydactylia syndromes.


Comparison of scintigraphy and magnetic resonance imaging for stress injuries of bone.
To compare findings of radiography, scintigraphy, and magnetic resonance imaging (MRI) in stress injuries of bone and evaluate changes of these findings with time correlated with clinical symptoms. Prospective study. A primary care hospital outpatient orthopedic clinic. All 31 patients with stress injuries of bone who visited our clinic from July 1996 to June 2000. Radiography, scintigraphy, MRI, and clinical examinations were performed on the same day or at least within 1 week of each other, and the findings were compared. If symptoms of stress injury of bone continued, these examinations were repeated at intervals of 2 months until symptoms disappeared. These radiologic findings were assessed by an independent radiologist who was blinded to the clinical symptoms of the patients. Correlation accuracy of MRI and scintigraphy findings with clinical symptoms. Even with negative initial radiographic findings, all initial scintigraphy and MRI indicated stress injury of bone. There were no patients with positive/negative examinations. Grade of scintigraphy and MRI were closely correlated, and these findings also correlated with the degree of clinical symptoms. Compared with scintigraphy, MRI showed more diagnostic information, such as fracture line and periosteal edema. Areas of increased activity in scintigraphy were consistent with the grades of MRI, especially high signal intensity areas of STIR (short tau inversion recovery) image. From these observations, MRI is less invasive, provides more information than scintigraphy, and is recommended for initial diagnosis and assessment stages of stress injury of bone.


Global, regional, and national time trends in mortality for stroke, 1990-2019: An age-period-cohort analysis for the global burden of disease 2019 study and implications for stroke prevention.
Despite the fact that stroke is the second leading cause of death globally, a comprehensive and comparable assessment of mortality, and epidemiologic trends has not been conducted for most regions.We estimated the global and regional burden of stroke from 1990 to 2019 using data from the 2019 Global Study of Diseases, Injuries, and Risk Factors. For the period between 1990 and 2019, we used an age-period-cohort model to calculate the annual percentage changes in mortality (net drifts), local drifts, and period and cohort relative risks (period/cohort effects). Meanwhile, to quantify the temporal trends in stroke age-standardised mortality rate (ASMR), Average annual percentage changes (AAPCs) were determined by sex, area. With the potential to uncover disparities and treatment gaps in stroke care, this approach enables the examination and differentiation of age, period, and cohort effects in mortality trends. Global stroke deaths in 2019 were 6,552,725 (95% UI 5,995,200 to 7,015,139). Between 1990 and 2019, the ASMR declined globally by 36.43% (95% UI -41.65 to -31.2), with decreases in all SDI quintiles. The net drift in stroke mortality from 1990 to 2019 varied from -2.83% per year (95% confidence interval [CI]:-3.39 to -2.77) in countries with a high Socio-demographic Index (SDI) to -1.21% per year (95% CI: -1.26 to -1.16) in countries with a low SDI. During the past 30 years, favorable mortality reductions were generally found in high-SDI countries (net drift = -3.1% [95% CI: -3.4 to -2.8] per year) and high-middle SDI countries (-2.8% [-3.0 to -2.6]). However, 31 of 204 countries had either increasing trends (net drifts≥0.0%) or stagnated reductions (≥ - 0.5%) in mortality. The relative risk of mortality generally showed improving trends over time and in successively younger birth cohorts among high and high-middle SDI countries, with the exceptions of Kuwait, Ukraine, Kazakhstan, Guam, RussianFederation, Lithuania, Turkey, Montenegro, Serbia, Bosnia and Herzegovin, and Bulgaria. Notwithstanding mortality from stroke has increased globally over the past 30 years, adverse period and cohort effects have been found in many countries, calling into question the adequacy of healthcare for stroke patients of all ages. These lapses have a significant impact on the likelihood of achieving the Sustainable Development Goal (SDG) targets on mortality from age 60+ and NCDs.


A report on the diagnosis of intermediate hyperglycemia in Korea: A pooled analysis of four community-based cohort studies.
Many studies show poor agreement between fasting plasma glucose (FPG)-based and 2-h postchallenge glucose (2-h PG)-based criteria to assess glucose metabolism. We examined the rate of agreement between FPG- and 2-h PG-based criteria in the diagnosis of intermediate hyperglycemia in four representative cohort studies in South Korea and compared the clinical characteristics and biochemical parameters in subjects with impaired fasting glucose (IFG) according to their FPG values. Of 6234 subjects from four population-based studies performed from 1993 to 2000, 4610 individuals with data from a 75g oral glucose tolerance test (OGTT) and no previous history of diabetes were selected. We examined the concordance rate between the FPG and 2-h PG-based criteria. We also investigated the differences in the clinical characteristics and biochemical parameters between individuals with IFG according to their FPG values. The fasting and 2-h PG criteria had large discordance rates in the diagnosis of diabetes and impaired glucose tolerance (IGT) in Korean adults. When individuals with IFG were classified into stage 1 [5.6-6.1mmol/L (100-109mg/dL)] and stage 2 [6.1-7.0mmol/L (110-125mg/dL)] IFG, individuals with stage 2 IFG are more obese and had higher blood pressure and total cholesterol and triglycerides concentrations compared with those with stage 1 IFG. In addition, more individuals with stage 2 IFG were with diabetes as determined by a 2-h PG>or=11.1mmol/L (14.1% vs. 1.9%) (P<0.05). Considering the poorer metabolic profile and higher percentage of people with diabetes by OGTT, these data indicate that, in the Korean population, individuals with stage 2 IFG should be treated differently from those with stage 1 IFG. To detect more cases of diabetes, the OGTT is recommended for all individuals with stage 2 IFG and cases with stage 1 IFG with some additional risk factors for diabetes.


FAP expression in alpha cells of Langherhans insulae-implications for FAPI radiopharmaceuticals' use.
Radiopharmaceuticals targeting fibroblast activation protein (FAP) alpha are increasingly studied for diagnostic and therapeutic applications. We discovered FAP expression at immunohistochemistry (IHC) in the alpha cells of the Langerhans insulae of few patients. Therefore, we planned an investigation aimed at describing FAP expression in the pancreas and discussing the implications for radioligand applications. We retrospectively included 40 patients from 2 institutions (20 pts each) according to the following inclusion/exclusion criteria: (i) pathology proven pancreatic ductal adenocarcinoma and neuroendocrine tumors (NET), 10 pts per each group at each center; (ii) and availability of paraffin-embedded tissue; and (iii) clinical-pathological records. We performed IHC analysis and applied a semiquantitative visual scoring system (0, negative staining; 1, present in less than 30%; 2, present in more than 30% of the area). FAP expression was assessed according to histology-NET (n = 20) vs ductal adenocarcinoma (n = 20)-and to previous treatments within the adenocarcinoma group. The local ethics committee approved the study (No. INT 21/16, 28 January 2016). The population consisted of 24 males and 16 females, with a median age of 68 and a range of 14-84 years; 8/20 adenocarcinoma patients received chemotherapy. In all the Langerhans insulae (40/40), pancreatic alpha cells were found to express FAP, with a score of 2. No difference was found among NET (20/20) and adenocarcinoma (20/20), nor according to neoadjuvant chemotherapy in the adenocarcinoma cohort (received or not received). Pancreatic Langerhans islet alpha cells normally express FAP. This is not expected to influence the diagnostic accuracy of FAP-targeting tracers. In the therapeutic setting, our results suggest the need to better elucidate FAPI radioligands' effects on the Langerhans insulae function.


Sustained vs. oscillating expressions of Ngn2, Dll1 and Hes1: a model of neural differentiation of embryonic telencephalon.
Neural progenitor cells show oscillatory expression of the Notch ligand Delta-like1 (Dll1), the Notch target Hes1 and the proneural gene Neurogenin 2 (Ngn2) during embryonic development of the mammalian telencephalon. On the other hand, expression of these genes is sustained in postmitotic neurons (upregulated for Ngn2 and Dll1, down regulated for Hes1). These facts suggest that a switch from oscillatory to sustained expression of proneural and other genes is critical in neural fate decisions. Moreover, despite controversies over the role of Numb in determining the neural fate in mammals, there is evidence that inheritance of Numb during neurogenic cell division is involved in neural differentiation. It is also known that mNumb activates Notch1 receptor degradation. The arrest of oscillations in a given cell may be due to increasing degradation of Notch1 brought about by mNumb during neurogenic division. We introduce a modification in a previous model of the gene network for two cells coupled by the Delta-Notch pathway (Wang et al., 2011). We analyze the consequences of an asymmetry between two neighbor cells in the rate of degradation of Notch (mimicking the effect of asymmetric inheritance of mNumb during the neurogenic division). The results show that a slight difference in Notch degradation between the two cells keeps oscillation going in one of them while oscillation stops in the other. Moreover, when Delta-Notch coupling is canceled, both cells show sustained expression (upregulated levels for Ngn2 and Dll1, downregulated for Hes1). We show that the model is stable against parameter variations. Moreover, to take into account the possible influence of the environment on both cells, neighboring cells are included in a mean field approximation. Both, parameter fluctuations and effects of the environment lead to asynchronous oscillations of Hes1/Ngn2 in different progenitor cells.


Stability studies of crude plant material of Bacopa monnieri and quantitative determination of bacopaside I and bacoside A by HPLC.
Bacopa monnieri (BM) contains several dammarane-type triterpenoid saponins including bacopaside I and bacoside A. These bioactive compounds may be used as chemical markers for the quality control of different BM products used for promoting mental health and intellect. Quantification of bacopaside I and bacoside A in crude plant material of BM stored under the stability study conditions by HPLC. Crude BM samples were stored at long-term (LS; 30°C and 65% RH), accelerated (AS; 40°C and 75% RH) and real-time (RT) study conditions. HPLC of BM extracts was carried out using a LiChroCART Purospher® STAR RP-18 endcapped column along with a guard column, Purospher STAR RP 18e 4.0 4.0 mm 5 µm using a gradient of acetonitrile (A) and water containing 0.05% (v/v) orthophosphoric acid (B) at a flow rate 1.5 mL/min with UV detection at 205 nm. The linear range of bacopaside I and bacoside A was 0.2 to 1 mg/mL. With the help of a regression equation the coefficient of determination (r²) values for bacopaside I and bacoside A were found to be > 0.999 and > 0.994 respectively. Relative standard deviation (RSD) values were < 4.0 for all the concentrations injected (n = 3). The HPLC study indicated that BM samples kept under LS condition are rich in saponin contents as compared with the samples stored under AS and RT study conditions. The study indicated that BM plant material should be used fresh to obtain maximum concentration of active saponins or it should be stored under LS conditions up to 3 months.


Simultaneous 31P- and 1H-nuclear magnetic resonance studies of hypoxia and ischemia in the cat brain.
The objective of this study was to evaluate simultaneous 31P/1H nuclear magnetic resonance (NMR) spectroscopy as a technique for monitoring and correlating changes in brain energy metabolism during hypoxia and ischemia. Five cats were studied with a protocol that involved 20 min of hypoxia (PaO2 20 mm), 60 min of recovery, 10 min of hypoxia with relative ischemia (bilateral carotid occlusion, PaO2 20 mm), and 60 min of recovery. Bifrontal and biparietal electrocorticograms (ECoG) were monitored continuously during the entire protocol. The results demonstrate that the degree of metabolic response is different in individual cats, but a number of quantitative relationships between metabolic parameters are consistently observed for all cats. First, there is agreement between increases in lactate and changes in intracellular pH; the observed relationship corresponds to an in vivo cerebral buffer capacity of 29 mumol/g/pH unit. Second, the delayed recovery of PCr is due to the effect of metabolic acidosis on the creatine kinase equilibrium and not to a delayed recovery of the ATP/ADP ratio. Third, the observed rate of lactate clearance from the cell is zero-order (k = 0.36 mumol/g/min) for lactate levels greater than 5 microns/g and may be composed of both lactate efflux from the cell and lactate oxidation.


Quasi-elastic light scattering of antigen-antibody complexes.
Many biological properties of immune complexes (IC) depend upon their size. Quasi-elastic light scattering (QLS) was used to measure a mean equivalent hydrodynamic radius (Rh) and variance of the distribution of model IC composed of bovine serum albumin (BSA) as antigen (Ag) and combinations of two or three well-characterized monoclonal antibodies (MAb) which bound noncompetitively to unique epitopes on BSA. With the molar ratio (X) of each MAb to Ag fixed, Rh increased with concn. Rh was maximal at equivalence (X = 0.5) with two MAb and at slight MAb excess (X = 0.67) with three MAb. The largest Rh with two MAb was about 200 A, and Rh was uniformly different amongst the three combinations of two MAb IC. The largest hydrodynamic radius of individual complexes which formed with two MAb was estimated to be about 400 A; even larger individual complexes were formed with three MAb. Size changes following alteration of solution concns were also followed with QLS. Kinetics of two MAb IC association were too fast to observe; dissociation following large dilution (40-fold) required 5-10 min to attain a new steady state, much less at small dilution. With three MAb, Rh dropped sharply in 5 min and became steady after 1-2 hr. These results suggest that conventional chromatographic and ultracentrifugation techniques for studying IC size, involving large dilution and long measurement time, provide misleading results. Association of three MAb produced a rapid initial increase of Rh in several min, followed by diverse behavior which depended upon concn. From high to low concn, these included (1) exponential growth of Rh with time and appearance of visible macroscopic particles; (2) metastable states for several hr followed by slow growth to large size over several days, leading to formation of particles; and (3) rapid growth to steady state conditions with no visible particles. This heretofore unobserved equilibrium and kinetic behavior of model IC in solution may be reflected in the behavior of more complex, naturally occurring IC.


Structure of peroxisomes and activity of the marker enzyme catalase in digestive epithelial cells in relation to PAH content of mussels from two Basque estuaries (Bay of Biscay): seasonal and site-specific variations.
The aim of the present work was to study the seasonal as well as the site-specific variations in the structure of peroxisomes and in the activity of the peroxisomal marker enzyme catalase in digestive epithelial cells of mussels to validate the potential use of these parameters as early biomarkers of environmental organic pollution in estuarine ecosystems. For this purpose, mussels were sampled monthly for 14 months in two Basque estuaries (Bay of Biscay) with different degrees of pollution. Stereological procedures were applied to detect changes in peroxisome structure, and microspectrophotometry was used to quantify changes in catalase activity. The animals from the two studied sampling sites presented differences in polycyclic aromatic hydrocarbon (PAH) burdens, mussels from Plentzia generally showing lower total PAH contents than mussels from Galea. The peroxisome structure of the animals from the two estuaries suffered seasonal variations that were of different kind and intensity in both sites. In this way, a strong peroxisome proliferatory response was found in mussels sampled in Plentzia during the summer months, while mussels from Galea presented few variations along the year. Catalase activity behaved similarly in the animals sampled in the two estuaries, with higher values in spring. It appeared that mussels exposed chronically to PAHs and other pollutants, such as those from Galea, lost their ability to respond to this exposure in terms of peroxisome proliferation. In contrast, mussels collected in Plentzia effectively responded to an increased bioavailability of organic pollutants during the summer by increasing peroxisome volume and surface and numerical densities in digestive epithelial cells. However, these increases were transient because elevated PAH body burdens detected in mussels sampled in Plentzia in autumn were not accompanied by a peroxisome proliferatory response. Further studies are needed before changes in peroxisomal structure and in the activity of catalase could be used as early biomarkers to assess environmental quality in pollution monitoring programs like the Mussel Watch.


Weight loss and modeled cost savings in a digital diabetes prevention program.
Participation in the National Diabetes Prevention Program (DPP) can improve individual health through reduced risk of type 2 diabetes and save the healthcare system substantial medical costs associated with a diagnosis of type 2 diabetes and its associated complications. There is less evidence of outcomes and cost savings associated with a fully digital delivery of the DPP. This study assessed 13,593 members who provided an initial digital weight and subsequently achieved various weight loss and engagement outcomes during their participation in a digital DPP. Analyzed data included both complete observations and missing observations imputed using maximum likelihood estimation. Findings include members' behavioral correlates of weight loss and a literature-based cost-savings estimate associated with achieving three mutually exclusive weight loss or engagement benchmarks: ≥5% weight loss, >2% but <5% weight loss, and completion of ≥4 educational lessons. 11,976 members (88%) provided a weight after 2 months of participation, enabling calculation of their weight nadir. Considering complete data, 97% of members maintained or lost weight. Using the imputed data for these calculations, 32.0% of members achieved ≥5%, 32.4% achieved >2% but <5%, 32.0% maintained ±2%, and 3.6% gained weight. Members who lost the most weight achieved their weight nadir furthest into the program (mean day = 189, SE = 1.4) and had the longest active engagement (mean days = 268, SE = 1.4), particularly compared to members who gained weight (mean nadir day = 119, SE = 3.7; active engagement mean days = 199, SE = 4.9) (both p ≤ 0.0001). Modeled 1-year cost-savings estimates ranged from $11,229,160 to $12,960,875. Members of a fully digital DPP achieved clinical and engagement outcomes during their participation in the program that confer important health benefits and cost savings.


Hypothalamic DNA methylation in rats with dihydrotestosterone-induced polycystic ovary syndrome: effects of low-frequency electro-acupuncture.
What is the central question of this study? What is the role of hypothalamic DNA methylation in the development of polycystic ovary syndrome (PCOS) and the response to electro-acupuncture treatment. What is the main finding and its importance? Global DNA methylation and expression of DNA methyltransferases (DNMTs) were increased in PCOS-like rats, and electro-acupuncture (EA) decreased global DNA methylation and DNMT3b expression. Pyrosequencing showed that the DNA methylation of some PCOS candidate genes was changed in the PCOS and PCOS+EA groups, suggesting that hypothalamic DNA methylation plays an important role in the development of PCOS and in mediating the effects of electro-acupuncture treatment. Polycystic ovary syndrome (PCOS) is a common reproductive and endocrine disease of unknown aetiology. Recently, epigenetic studies focusing on DNA methylation in PCOS have received much attention, but the mechanisms are still unclear. In the present study, we used the 5α-dihydrotestosterone-induced PCOS-like rat model and treated the rats with electro-acupuncture (EA). Rats were randomly divided into four groups - controls, diet-induced obesity, PCOS and PCOS+EA. We examined the reproductive, metabolic and behavioural phenotypes, validated the effect of EA, and explored the role of hypothalamic DNA methylation by analysing the methylation of global DNA and selected candidate genes. The PCOS rats presented with reproductive dysfunctions such as lack of regular oestrous cyclicity, metabolic disorders such as increased body weight and insulin resistance, and depression and anxiety-like behaviours. EA improved the reproductive functions, decreased body weight and improved experimental depressive behaviour. Furthermore, global DNA methylation and the expression of DNA methyltransferases (DNMTs) were increased in PCOS rats compared to the control group, and EA decreased the global DNA methylation and the expression of DNMT3b. In addition, pyrosequencing showed that the DNA methylation of certain CpG sites in targeted genes (Plcg1, Camk2b, Esr2 and Pgr) was increased in the PCOS group, but the DNA methylation of Camk2b and Ar was decreased after EA treatment. These results indicate that hypothalamic DNA methylation might be correlated with the development of PCOS and that EA has an effect on hypothalamic DNA methylation in PCOS rats.


Circulating anti-C3b IgG in lupus nephritis: A large cohort study.
The current study aimed to analyze the clinical significance and bio-functional properties of anti-C3b IgG based on a lupus nephritis cohort. We found that the prevalence of anti-C3b IgG in our cohort was 47.8%. Patients with positive anti-C3b IgG had significantly higher SLEDAI, lower circulating C3 and C4 levels. Anti-C3b IgG levels were positively correlated with C3 or C1q deposition in kidneys and several active pathological lesions. The positivity of anti-C3b IgG was an independent risk factor for the composite endpoints in the subgroup of proliferative lupus nephritis patients. In vitro, the purified IgG fractions from positive patients resulted in increased C3a generation through the alternative pathway, and interfered factor H and CR1 binding to C3b. Our findings indicated that anti-C3b IgG associated with local renal injury and long-term outcomes in lupus nephritis patients, possibly through leading to the complement alternative pathway over-activation.