Critical reduction in beta-cell mass results in two distinct outcomes over time. Adaptation with impaired glucose tolerance or decompensated diabetes.
We have proposed that hyperglycemia-induced dedifferentiation of beta-cells is a critical factor for the loss of insulin secretory function in diabetes. Here we examined the effects of the duration of hyperglycemia on gene expression in islets of partially pancreatectomized (Px) rats. Islets were isolated, and mRNA was extracted from rats 4 and 14 weeks after Px or sham Px surgery. Px rats developed different degrees of hyperglycemia; low hyperglycemia was assigned to Px rats with fed blood glucose levels less than 150 mg/dl, and high hyperglycemia was assigned above 150 mg/dl. beta-Cell hypertrophy was present at both 4 and 14 weeks. At the same time points, high hyperglycemia rats showed a global alteration in gene expression with decreased mRNA for insulin, IAPP, islet-associated transcription factors (pancreatic and duodenal homeobox-1, BETA2/NeuroD, Nkx6.1, and hepatocyte nuclear factor 1 alpha), beta-cell metabolic enzymes (glucose transporter 2, glucokinase, mitochondrial glycerol phosphate dehydrogenase, and pyruvate carboxylase), and ion channels/pumps (Kir6.2, VDCC beta, and sarcoplasmic reticulum Ca(2+)-ATPase 3). Conversely, genes normally suppressed in beta-cells, such as lactate dehydrogenase-A, hexokinase I, glucose-6-phosphatase, stress genes (heme oxygenase-1, A20, and Fas), and the transcription factor c-Myc, were markedly increased. In contrast, gene expression in low hyperglycemia rats was only minimally changed at 4 weeks but significantly changed at 14 weeks, indicating that even low levels of hyperglycemia induce beta-cell dedifferentiation over time. In addition, whereas 2 weeks of correction of hyperglycemia completely reverses the changes in gene expression of Px rats at 4 weeks, the changes at 14 weeks were only partially reversed, indicating that the phenotype becomes resistant to reversal in the long term. In conclusion, chronic hyperglycemia induces a progressive loss of beta-cell phenotype with decreased expression of beta-cell-associated genes and increased expression of normally suppressed genes, these changes being present with even minimal levels of hyperglycemia. Thus, both the severity and duration of hyperglycemia appear to contribute to the deterioration of the beta-cell phenotype found in diabetes.


Responses to prolactin secretagogues in oestrogen-treated rats suggest that the defect in prolactin regulation produced by oestrogen is at the level of the pituitary gland.
Prolactin responses to pharmacological agents were used to characterize the defect in prolactin regulation which occurs after administration of high doses of oestrogen to rats. Animals with chronically implanted venous cannulae were injected with 2 mg oestradiol benzoate in oil and 2-3 days later prolactin concentrations were measured after injections of saline, thyrotrophin-releasing hormone (TRH), fenfluramine, apomorphine and butaclamol. The responses were compared with those in oil-injected animals. Hyperprolactinaemia in oestrogen-treated animals was unresponsive to apomorphine, but was even more sensitive to dopamine receptor blockade than controls. These results suggest that the lactotrophs in oestrogen-treated animals are already maximally suppressed by endogenous dopamine, though ineffectively. Although there was an increased prolactin response to TRH in oestrogen-treated animals, there was an impaired response to fenfluramine, indicating suppressed serotonergic prolactin-releasing factor mechanisms. Maximal endogenous dopaminergic activity and suppressed prolactin-releasing factor mechanisms are appropriate hypothalamic responses to hyperprolactinaemia. The operation of these responses in the earliest stages of the development of pituitary hyperplasia indicates that oestrogen induces a disturbance of prolactin regulation in the lactotroph, independent of hypothalamic control.


Nanomolar Protein Thermal Profiling with Modified Cyanine Dyes.
Protein properties and interactions have been widely investigated by using external labels. However, the micromolar sensitivity of the current dyes limits their applicability due to the high material consumption and assay cost. In response to this challenge, we synthesized a series of cyanine5 (Cy5) dye-based quencher molecules to develop an external dye technique to probe proteins at the nanomolar protein level in a high-throughput one-step assay format. Several families of Cy5 dye-based quenchers with ring and/or side-chain modifications were designed and synthesized by introducing organic small molecules or peptides. Our results showed that steric hindrance and electrostatic interactions are more important than hydrophobicity in the interaction between the luminescent negatively charged europium-chelate-labeled peptide (Eu-probe) and the quencher molecules. The presence of substituents on the quencher indolenine rings reduces their quenching property, whereas the increased positive charge on the indolenine side chain improved the interaction between the quenchers and the luminescent compound. The designed quencher structures entirely altered the dynamics of the Eu-probe (protein-probe) for studying protein stability and interactions, as we were able to reduce the quencher concentration 100-fold. Moreover, the new quencher molecules allowed us to conduct the experiments using neutral buffer conditions, known as the peptide-probe assay. These improvements enabled us to apply the method in a one-step format for nanomolar protein-ligand interaction and protein profiling studies instead of the previously developed two-step protocol. These improvements provide a faster and simpler method with lower material consumption.


Distinct mesoderm migration phenotypes in extra-embryonic and embryonic regions of the early mouse embryo.
In mouse embryo gastrulation, epiblast cells delaminate at the primitive streak to form mesoderm and definitive endoderm, through an epithelial-mesenchymal transition. Mosaic expression of a membrane reporter in nascent mesoderm enabled recording cell shape and trajectory through live imaging. Upon leaving the streak, cells changed shape and extended protrusions of distinct size and abundance depending on the neighboring germ layer, as well as the region of the embryo. Embryonic trajectories were meandrous but directional, while extra-embryonic mesoderm cells showed little net displacement. Embryonic and extra-embryonic mesoderm transcriptomes highlighted distinct guidance, cytoskeleton, adhesion, and extracellular matrix signatures. Specifically, intermediate filaments were highly expressed in extra-embryonic mesoderm, while live imaging for F-actin showed abundance of actin filaments in embryonic mesoderm only. Accordingly, Rhoa or Rac1 conditional deletion in mesoderm inhibited embryonic, but not extra-embryonic mesoderm migration. Overall, this indicates separate cytoskeleton regulation coordinating the morphology and migration of mesoderm subpopulations.


Predictors and outcomes of frequent emergency department users.
To identify predictors and outcomes associated with frequent emergency department (ED) users. Cross-sectional intake surveys, medical chart reviews, and telephone follow-up interviews of patients presenting with selected chief complaints were performed at five urban EDs during a one-month study period in 1995. Frequent use was defined by four or more self-reported, prior ED visits. Multivariate logistic regression identified predictors of frequent ED visitors from five domains (demographics, health status, health access, health care preference, and severity of acute illness). Associations between high use and selected outcomes were assessed with logistic regression models. All study components were completed by 2,333 of 3,455 eligible patients (67.5%). Demographics predicting frequent use included being a single parent, single or divorced marital status, high school education or less, and income of less than $10,000 (1995). Health status predictors included hospitalization in the preceding three months, high ratings of psychological distress, and asthma. Health access predictors included identifying an ED or a hospital clinic as the primary care site, having a primary care physician (PCP), and visiting a PCP in the past month. Choosing the ED for free care was the only health preference predictive of heavy use. Illness severity measures were higher in frequent visitors, although these were not independently predictive in the multivariate model. Outcomes correlated with heavy use include increased hospital admissions, higher rates of ED return visits, and lower patient satisfaction, but not willingness to return to the ED or follow-up with a doctor. Frequent ED visits are associated with socioeconomic distress, chronic illness, and high use of other health resources. Efforts to reduce ED visits require addressing the unique needs of these patients in the emergency and primary care settings.


A robotic platform to assess, guide and perturb rat forelimb movements.
Animal models are widely used to explore the mechanisms underlying sensorimotor control and learning. However, current experimental paradigms allow only limited control over task difficulty and cannot provide detailed information on forelimb kinematics and dynamics. Here we propose a novel robotic device for use in motor learning investigations with rats. The compact, highly transparent, three degree-of-freedom manipulandum is capable of rendering nominal forces of 2 N to guide or perturb rat forelimb movements, while providing objective and quantitative assessments of endpoint motor performance in a 50×30 mm(2) planar workspace. Preliminary experiments with six healthy rats show that the animals can be familiarized with the experimental setup and are able to grasp and manipulate the end-effector of the robot. Further, dynamic perturbations and guiding force fields (i.e., haptic tunnels) rendered by the device had significant influence on rat motor behavior (ANOVA, ). This approach opens up new research avenues for future characterizations of motor learning stages, both in healthy and in stroke models.


The assessment of psychological factors on upper extremity disability: A scoping review.
The primary purpose of this scoping review was to describe the nature and extent of the published research that assesses the relationship between psychological features and patient-reported outcome following surgery or rehabilitation of upper extremity disease or injury. Twenty-two included studies were examined for quantitative study design, outcome measure, inclusion/exclusion criteria, follow-up and recruitment strategy. Patient population and psychological assessment tools were examined for validity. Twenty-two studies met the inclusion criteria for this study. Only 7 of the 22 studies were longitudinal and the rest were cross sectional studies. Depression was the most common psychological status of interest and was included in 17 studies. Pain catastrophizing was the psychological status of interest in 5 of the studies. Four studies considered anxiety, 3 considered pain anxiety, 3 considered distress, 2 considered coping, 2 considered catastrophic thinking, and 2 considered fear avoidance beliefs. The majority of studies in this review were cross-sectional studies. Cross-sectional studies may not provide conclusive information about cause-and-effect relationships. This review encourages clinicians to be mindful of the psychological implications found in rehabilitation of individuals with upper extremity disease or injury along with being cognizant of choosing appropriate measurement tools that best represent each patient's characteristics and diagnoses. The nature of the research addressing psychological factors affecting outcomes after hand injury focus on negative traits and have limited strength to suggest causation as most have used cross-sectional designs. Stronger longitudinal designs and consideration of positive traits are needed in future studies.


Anti-inflammatory potential of total saponins derived from the roots of Panax ginseng in lipopolysaccharide-activated RAW 264.7 macrophages.
Ginseng, the root of Panax ginseng C.A. Meyer (Araliaceae), is a widely known traditional medicine that has been utilized throughout Asia for several thousand years. Ginseng saponins exert various important pharmacological effects regarding the control of a number of diseases. The aim of the present study was to identify the anti-inflammatory effects of total saponins extracted from ginseng (TSG) on lipopolysaccharide (LPS)-stimulated mouse RAW 264.7 macrophages. The inhibitory effects of TSG on LPS-induced nitric oxide (NO) production and LPS-induced tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) protein expression were determined by measuring the levels of nitrite and enzyme-linked immunosorbent assays, respectively. Furthermore, the effects of TSG on the mRNA expression levels and localizations of inducible NO synthase (iNOS), IL-1β and TNF-α, and their upstream signaling proteins, including nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs), were investigated by reverse transcription-polymerase chain reaction and western blotting, respectively. Following stimulation with LPS, elevated levels of NO production were detected in RAW 264.7 cells; however, TSG pretreatment significantly inhibited the production of NO (P<0.05), by suppressing the expression of iNOS. In addition, LPS-stimulated TNF-α and IL-1β production was significantly reduced by TSG (P<0.05). In the LPS-stimulated RAW 264.7 cells, NF-κB was translocated from the cytosol to the nucleus, whilst TSG pretreatment induced the sequestration of NF-κB in the cytosol by inhibiting inhibitor of κB degradation. TSG also contributed to downregulation of MAPKs in LPS-stimulated RAW 264.7 cells. These results suggested that TSG may exert anti-inflammatory activity, and that TSG may be considered a potential therapeutic for the treatment of inflammatory diseases associated with macrophage activation.


Revertant mosaicism in a human skin fragility disorder results from slipped mispairing and mitotic recombination.
Spontaneous gene repair, also called revertant mosaicism, has been documented in several genetic disorders involving organs that undergo self-regeneration, including the skin. Genetic reversion may occur through different mechanisms, and in a single individual, the mutation can be repaired in various ways. Here we describe a disseminated pattern of revertant mosaicism observed in 6 patients with Kindler syndrome (KS), a genodermatosis caused by loss of kindlin-1 (encoded by FERMT1) and clinically characterized by patchy skin pigmentation and atrophy. All patients presented duplication mutations (c.456dupA and c.676dupC) in FERMT1, and slipped mispairing in direct nucleotide repeats was identified as the reversion mechanism in all investigated revertant skin spots. The sequence around the mutations demonstrated high propensity to mutations, favoring both microinsertions and microdeletions. Additionally, in some revertant patches, mitotic recombination generated areas with homozygous normal keratinocytes. Restoration of kindlin-1 expression led to clinically and structurally normal skin. Since loss of kindlin-1 severely impairs keratinocyte proliferation, we predict that revertant cells have a selective advantage that allows their clonal expansion and, consequently, the improvement of the skin condition.


Robust model predictive control of nonlinear systems with unmodeled dynamics and bounded uncertainties based on neural networks.
This paper presents a neural network approach to robust model predictive control (MPC) for constrained discrete-time nonlinear systems with unmodeled dynamics affected by bounded uncertainties. The exact nonlinear model of underlying process is not precisely known, but a partially known nominal model is available. This partially known nonlinear model is first decomposed to an affine term plus an unknown high-order term via Jacobian linearization. The linearization residue combined with unmodeled dynamics is then modeled using an extreme learning machine via supervised learning. The minimax methodology is exploited to deal with bounded uncertainties. The minimax optimization problem is reformulated as a convex minimization problem and is iteratively solved by a two-layer recurrent neural network. The proposed neurodynamic approach to nonlinear MPC improves the computational efficiency and sheds a light for real-time implementability of MPC technology. Simulation results are provided to substantiate the effectiveness and characteristics of the proposed approach.


Gulf war depleted uranium risks.
US and British forces used depleted uranium (DU) in armor-piercing rounds to disable enemy tanks during the Gulf and Balkan Wars. Uranium particulate is generated by DU shell impact and particulate entrained in air may be inhaled or ingested by troops and nearby civilian populations. As uranium is slightly radioactive and chemically toxic, a number of critics have asserted that DU exposure has resulted in a variety of adverse health effects for exposed veterans and nearby civilian populations. The study described in this paper used mathematical modeling to estimate health risks from exposure to DU during the 1991 Gulf War for both US troops and nearby Iraqi civilians. The analysis found that the risks of DU-induced leukemia or birth defects are far too small to result in an observable increase in these health effects among exposed veterans or Iraqi civilians. The analysis indicated that only a few ( approximately 5) US veterans in vehicles accidentally targeted by US tanks received significant exposure levels, resulting in about a 1.4% lifetime risk of DU radiation-induced fatal cancer (compared with about a 24% risk of a fatal cancer from all other causes). These veterans may have also experienced temporary kidney damage. Iraqi children playing for 500 h in DU-destroyed vehicles are predicted to incur a cancer risk of about 0.4%. In vitro and animal tests suggest the possibility of chemically induced health effects from DU internalization, such as immune system impairment. Further study is needed to determine the applicability of these findings for Gulf War exposure to DU. Veterans and civilians who did not occupy DU-contaminated vehicles are unlikely to have internalized quantities of DU significantly in excess of normal internalization of natural uranium from the environment.


Three-dimensional mapping of gallbladder wall thickness on computed tomography using Laplace's equation.
Traditionally, maximum gallbladder wall thickness is measured at a single point on ultrasonography. The purpose of this work was to develop an automated technique to measure the thickness of the gallbladder wall over the entire gallbladder surface using computer tomography (CT). Subjects who had (5-mm) thick and thin (2.5-mm) reconstruction through the abdomen were selected from a research database. Their volumetric computed tomographic images were acquired using a multidetector GE Medical Systems LightSpeed 16 scanner at 120 kVp, approximately 250 mAs, with standard filter reconstruction algorithm and segmented in three dimensions. Two segmentation boundaries were obtained, an inner and an outer boundary of the gallbladder wall. The thickness of the wall was quantified by computing the distance between the boundaries over the entire volume using Laplace's equation from mathematical physics. The distance between the surfaces is found by computing normalized gradients that form a vector field, representing tangent vectors along field lines connecting both boundaries. The Laplacian technique was compared with the well-known Euclidean distance transformation (EDT) technique that provides a three-dimensional Euclidean distance mapping between the two extracted surfaces. The technique was tested on 10 subjects who had thin- and thick-section computed tomographic datasets reconstructed from a single scan. The mean thickness for the thick- and thin-section CT using Laplace was 3.18 and 2.93 mm, respectively. The smooth transition between surfaces resulting from the Laplace technique resulted in a coefficient of variation that was less than 1% compared to EDT. EDT technique is very sensitive to imperfect segmentations, resulting in higher variation compared to the Laplacian technique. The smooth transition between surfaces makes the Laplacian technique more robust compared to EDT for the measurement of CT gallbladder thickness.


MHC class II invariant chain-adjuvanted viral vectored vaccines enhances T cell responses in humans.
Strategies to enhance the induction of high magnitude T cell responses through vaccination are urgently needed. Major histocompatibility complex (MHC) class II-associated invariant chain (Ii) plays a critical role in antigen presentation, forming MHC class II peptide complexes for the generation of CD4+ T cell responses. Preclinical studies evaluating the fusion of Ii to antigens encoded in vector delivery systems have shown that this strategy may enhance T cell immune responses to the encoded antigen. We now assess this strategy in humans, using chimpanzee adenovirus 3 and modified vaccinia Ankara vectors encoding human Ii fused to the nonstructural (NS) antigens of hepatitis C virus (HCV) in a heterologous prime/boost regimen. Vaccination was well tolerated and enhanced the peak magnitude, breadth, and proliferative capacity of anti-HCV T cell responses compared to non-Ii vaccines in humans. Very high frequencies of HCV-specific T cells were elicited in humans. Polyfunctional HCV-specific CD8+ and CD4+ responses were induced with up to 30% of CD3+CD8+ cells targeting single HCV epitopes; these were mostly effector memory cells with a high proportion expressing T cell activation and cytolytic markers. No volunteers developed anti-Ii T cell or antibody responses. Using a mouse model and in vitro experiments, we show that Ii fused to NS increases HCV immune responses through enhanced ubiquitination and proteasomal degradation. This strategy could be used to develop more potent HCV vaccines that may contribute to the HCV elimination targets and paves the way for developing class II Ii vaccines against cancer and other infections.


A Classification Method for Seed Viability Assessment with Infrared Thermography.
This paper presents a viability assessment method for Pisum sativum L. seeds based on the infrared thermography technique. In this work, different artificial treatments were conducted to prepare seeds samples with different viability. Thermal images and visible images were recorded every five minutes during the standard five day germination test. After the test, the root length of each sample was measured, which can be used as the viability index of that seed. Each individual seed area in the visible images was segmented with an edge detection method, and the average temperature of the corresponding area in the infrared images was calculated as the representative temperature for this seed at that time. The temperature curve of each seed during germination was plotted. Thirteen characteristic parameters extracted from the temperature curve were analyzed to show the difference of the temperature fluctuations between the seeds samples with different viability. With above parameters, support vector machine (SVM) was used to classify the seed samples into three categories: viable, aged and dead according to the root length, the classification accuracy rate was 95%. On this basis, with the temperature data of only the first three hours during the germination, another SVM model was proposed to classify the seed samples, and the accuracy rate was about 91.67%. From these experimental results, it can be seen that infrared thermography can be applied for the prediction of seed viability, based on the SVM algorithm.


Physical Function, Physical Activity, and Quality of Life After Liver Transplantation.
Robust physical activity after liver transplantation is an important determinant of longterm health, similar in its importance to the value of pretransplant activity for withstanding the immediate stress of transplantation. Although transplantation normally enables rapid recovery of liver synthetic and metabolic functions, the recovery of physical capacity and performance to normal levels is delayed and often incomplete. Anatomic measurements of sarcopenia and the physical performance indicators of frailty both tend to improve slowly, and they may, in fact, decrease further in the posttransplant period, especially when the common extrahepatic drivers of muscle loss, such as the elements of the metabolic syndrome, persist or intensify after transplantation. Posttransplant exercise improves fitness, which is a conclusion based on 2 observational studies and 3 randomized trials that assessed endpoints of strength testing, energy expenditure in metabolic equivalents, and peak or maximal oxygen uptake. Importantly, 1 controlled trial found that exercise also improved quality of life (QOL) measured by the Short Form 36 survey, consistent with multiple reports of the value of social support and engagement in sports activity for improving posttransplant QOL. Developing evidence-based standards for post-liver transplant physical activity baseline testing and sustainment of intensity and quality is a key unmet need in transplant hepatology. At present, it is reasonable for transplant teams to assess fitness and design a tailored exercise program when a recipient is first discharged, to record and reinforce progress at all posttransplant visits, and to set realistic longterm performance goals that will often achieve recommended standards for the healthy general population.


Use of functional MRI to guide decisions in a clinical stroke trial.
An investigational trial examined safety and efficacy of targeted subthreshold cortical stimulation in patients with chronic stroke. The anatomical location for the target, hand motor area, varies across subjects, and so was localized with functional MRI (fMRI). This report describes the experience of incorporating standardized fMRI into a multisite stroke trial. At 3 enrollment centers, patients moved (0.25 Hz) the affected hand during fMRI. Hand motor function was localized at a fourth center guiding intervention for those randomized to stimulation. The fMRI results were available within 24 hours. Across 12 patients, activation site variability was substantial (12, 23, and 11 mm in x, y, and z directions), exceeding stimulating electrode dimensions. Use of fMRI to guide decision-making in a clinical stroke trial is feasible.


CRISPR/Cas9-based gene activation and base editing in Populus.
The genus Populus has long been used for environmental, agroforestry and industrial applications worldwide. Today Populus is also recognized as a desirable crop for biofuel production and a model tree for physiological and ecological research. As such, various modern biotechnologies, including CRISPR/Cas9-based techniques, have been actively applied to Populus for genetic and genomic improvements for traits such as increased growth rate and tailored lignin composition. However, CRISPR/Cas9 has been primarily used as the active Cas9 form to create knockouts in the hybrid poplar clone "717-1B4" (P. tremula x P. alba clone INRA 717-1B4). Alternative CRISPR/Cas9-based technologies, e.g. those involving modified Cas9 for gene activation and base editing, have not been evaluated in most Populus species for their efficacy. Here we employed a deactivated Cas9 (dCas9)-based CRISPR activation (CRISPRa) technique to fine-tune the expression of two target genes, TPX2 and LecRLK-G which play important roles in plant growth and defense response, in hybrid poplar clone "717-1B4" and poplar clone "WV94" (P. deltoides "WV94"), respectively. We observed that CRISPRa resulted in 1.2-fold to 7.0-fold increase in target gene expression through transient expression in protoplasts and Agrobacterium-mediated stable transformation, demonstrating the effectiveness of dCas9-based CRISPRa system in Populus. In addition, we applied Cas9 nickase (nCas9)-based cytosine base editor (CBE) to precisely introduce premature stop codons via C-to-T conversion, with an efficiency of 13%-14%, in the target gene PLATZ which encodes a transcription factor involved in plant fungal pathogen response in hybrid poplar clone "717-1B4". Overall, we showcase the successful application of CRISPR/Cas-based technologies in gene expression regulation and precise gene engineering in two Populus species, facilitating the adoption of emerging genome editing tools in woody species.


[Cataract blindness in people 50 years old or older in a semirural area of northern Peru].
To determine the prevalence of blindness and of cataract blindness in persons 50 years of age or older in Piura and Tumbes, which are two departments in northern Peru, and to describe the coverage and quality of cataract surgery services in that area, and the barriers that prevent access to those services. Systematic sampling of persons 50 years old or older was done in Piura and Tumbes between August 2002 and March 2003, with 80 clusters of 60 people each being selected. Of the 4 800 persons chosen, 4,782 of them were examined, using a survey instrument that gathered general information on each person, the results of the visual acuity test and the lens examination, and information on cataract surgery or why that surgery had not been done. Visual acuity (VA) testing was done with a Snellen optotype with the letter "E," with sizes of 20/60 and 20/200 at distances of 6 m and 3 m, respectively, with the person using the visual correction (glasses) available. When the VA was less than 20/60 in one of the eyes, vision was tested with pinhole glasses. The prevalence of bilateral blindness (VA < 20/400) due to cataract, adjusted by age and sex, was 2.1% (95% confidence interval (CI): 1.7% to 2.6%). Among the 193 persons with bilateral blindness due to any cause, cataract was the cause in 104 of them (53.9%). The prevalence of blindness due to cataract or other causes increased with age and was higher in women than in men. Only 25% of the persons studied who needed cataract surgery had had that done. The prevalence of bilateral VA less than 20/200 due to unoperated cataract was 6.3% (95% CI: 5.3% to 7.3%); only 12% of the persons with that level of visual deficiency had had cataract surgery. The VA of 26% of the eyes operated on for cataract was lower than 20/200. The reasons given by persons who needed cataract surgery but who had not had it included not being able to pay for the operation (28%), lack of knowledge concerning cataracts (25%), fear of the operation (23%), and fear of completely losing their sight (17%). Most of the cases of blindness and of serious deficiency in visual acuity in persons 50 years old or older in Piura and Tumbes are due to uncorrected refractive defects, especially cataracts. The high prevalence of bilateral blindness due to cataracts (2.1%) could be reduced with measures that facilitate access to appropriate treatment.


Fetal growth restriction induced by chronic placental insufficiency has long-term effects on the retina but not the optic nerve.
Reduced birth weight is associated with an increased risk of visual impairments. This study was undertaken to determine whether prenatal exposure to a chronic compromise sufficient to cause fetal growth restriction (FGR) results in long-term alterations to the retina and optic nerve. FGR was induced by umbilicoplacental embolization (UPE) in two cohorts of pregnant ewes from (1) 120 days of gestation (dg) until 140 dg and (2) 120 dg until term ( approximately 147 dg). Control fetuses were not subjected to UPE. The structure and neurochemistry of the retina and number and structure of ganglion cell axons were assessed in near-term (140 dg) and adult animals (2.3 years). In near-term FGR fetuses compared with control fetuses there were significant reductions (P < 0.05) in the outer plexiform layer (OPL), the photoreceptor inner and outer segment layers, the inner nuclear layer (INL) in the central retina and the outer nuclear layer (ONL) in the peripheral retina, and the diameter of ganglion cell axons in the optic nerve, with a proportional reduction in the thickness of myelin sheaths. In FGR animals compared with the control at 2.3 years, there were significant reductions (P < 0.05) in the total thickness of the retina, the thickness of the photoreceptor outer segment layer and the INL and the number of tyrosine hydroxylase-immunoreactive (TH-IR) dopaminergic amacrine cells. Axonal diameter and myelin sheath thickness in the optic nerve were not different (P > 0.05) between groups. Chronic placental insufficiency in late gestation results in long-lasting effects on specific retinal components, including photoreceptor outer segments and TH-IR amacrine cells. Other alterations observed at term, including reductions in growth and myelination of optic nerve axons, do not persist, suggesting delayed rather than permanently compromised development. Alterations persisting into adulthood could affect visual function.


Temporal patterns of ant diversity across a mountain with climatically contrasting aspects in the tropics of Africa.
Factors that drive species richness over space and time are still poorly understood and are often context specific. Identifying these drivers for ant diversity has become particularly relevant within the context of contemporary global change events. We report on a long-term bi-annual (wet and dry seasons), standardized sampling of epigeal ants over a five year period on the mesic and arid aspects of an inselberg (Soutpansberg Mountain Range) in the tropics of Africa. We detail seasonal, annual and long-term trends of species density, test the relative contribution of geometric constraints, energy, available area, climate, local environmental variables, time, and space in explaining ant species density patterns through Generalized Linear Mixed Models (GLMM) where replicates were included as random factors to account for temporal pseudo-replication. Seasonal patterns were very variable and we found evidence of decreased seasonal variation in species density with increased elevation. The extent and significance of a decrease in species density with increased elevation varied with season. Annual patterns point to an increase in ant diversity over time. Ant density patterns were positively correlated with mean monthly temperature but geometric constraints dominated model performance while soil characteristics were minor correlates. These drivers and correlates accounted for all the spatio-temporal variability in the database. Ant diversity was therefore mainly determined by geometric constraints and temperature while soil characteristics (clay and carbon content) accounted for smaller but significant amounts of variation. This study documents the role of season, elevation and their interaction in affecting ant species densities while highlighting the importance of neutral processes and temperature in driving these patterns.


Lipidomics of Human Blood Plasma by High-Resolution Shotgun Mass Spectrometry.
Clinical lipidomics is an emerging biomarker discovery approach that compares lipid profiles under pathologically and physiologically normal conditions. Here we describe a method for the absolute (molar) quantification of more than 200 molecules from 14 major lipid classes from 5 μL of human blood plasma using high-resolution top-down shotgun mass spectrometry. Because of its technical simplicity and robustness, the protocol lends itself for high-throughput clinical lipidomics screens.


Chinese Public's Engagement in Preventive and Intervening Health Behaviors During the Early Breakout of COVID-19: Cross-Sectional Study.
Since January 2020, the coronavirus disease (COVID-19) swept over China and then the world, causing a global public health crisis. People's adoption of preventive and intervening behaviors is critical in curbing the spread of the virus. The aim of this study is to evaluate Chinese people's adoption of health behaviors in responding to COVID-19 and to identify key determinants for their engagement. An anonymous online questionnaire was distributed in early February 2020 among Mainland Chinese (18 years or older) to examine their engagement in preventive behaviors (eg, frequent handwashing, wearing masks, staying at home) and intervening behaviors (eg, advising family to wash hands frequently), and to explore potential determinants for their adoption of these health behaviors. Out of 2949 participants, 55.3% (n=1629) reported frequent engagement in preventive health behaviors, and over 84% (n=2493) performed at least one intervening health behavior. Greater engagement in preventive behaviors was found among participants who received higher education, were married, reported fewer barriers and greater benefits of engagement, reported greater self-efficacy and emotional support, had greater patient-centered communication before, had a greater media literacy level, and had greater new media and traditional media use for COVID-19 news. Greater engagement in intervening behaviors was observed among participants who were married, had lower income, reported greater benefits of health behaviors, had greater patient-centered communication before, had a lower media literacy level, and had a greater new media and traditional media use for COVID-19 news. Participants' engagement in coronavirus-related preventive and intervening behaviors was overall high, and the associations varied across demographic and psychosocial variables. Hence, customized health interventions that address the determinants for health behaviors are needed to improve people's adherence to coronavirus-related behavior guidelines.


Endocytosis is required for the growth of vacuolar H(+)-ATPase-defective yeast: identification of six new END genes.
Yeast mutants that are defective in acidification of the lysosome-like vacuole are able to grow at pH 5.5, but not at pH 7. Here, we present evidence that endocytosis is required for this low pH-dependent growth and use this observation to develop a screen for mutants defective in endocytosis. By isolating mutants that cannot grow when they lack the 60-kD vacuolar ATPase subunit (encoded by the VAT2 gene), we isolated a number of vat2-synthetic lethal (Vsl-) mutant strains. Seven of the Vsl- mutants are defective in endocytosis. Four of these mutant strains (end8-1, end9-1, end10-1, and end11-1) show altered uptake of the endocytosed ligand, alpha-factor, and three (end12-1, end12-2, and end13-1) are probably defective in transfer of internalized material to the vacuole. Most of the mutations also confer a strong Ts- growth defect. The mutants defective in uptake of alpha-factor sort newly synthesized vacuolar proteins correctly, while those which may be defective in subsequent transport steps secrete at least a fraction of the newly synthesized soluble vacuolar proteins. The mutations that result in a defect in alpha-factor uptake are not allelic to any of the genes previously shown to encode endocytic functions.


Postoperative antibiotic prophylaxis in total hip and knee arthroplasty: a systematic review and meta-analysis of randomized controlled trials.
Postoperative antibiotic prophylaxis is currently the standard of care for patients undergoing total hip and knee arthroplasty. We evaluated the evidence for this practice in the reduction of surgical-site infections. We systematically searched MEDLINE, Embase and the Cochrane Library for randomized controlled trials (RCTs) published up to Aug. 15, 2014. We included all RCTs that compared postoperative antibiotic prophylaxis with postoperative placebo or no treatment in patients undergoing primary total hip or knee arthroplasty for osteoarthritis. We combined outcomes for surgical-site infection using a random-effects model and quantified heterogeneity using the χ2 test and the I2 statistic. We assessed the overall quality of the evidence according to the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach. We identified 4 RCTs (n = 4036) that met the inclusion criteria. Surgical-site infections occurred in 3.1% (63/2055) of patients in the prophylaxis group and 2.3% (45/1981) in the control group. Postoperative prophylaxis did not reduce the rate of surgical-site infections compared with placebo (risk difference 0.01, 95% confidence interval 0.00 to 0.02; I2 = 26%). This result was robust to sensitivity testing for losses to follow-up. According to the GRADE approach, the overall quality of evidence was very low. The available evidence did not show efficacy of postoperative antibiotic prophylaxis for the prevention of surgical-site infections in patients undergoing total hip or knee arthroplasty. Multicentred RCTs are likely to have an important impact on the confidence in the effect estimate and to change the estimate itself.


Impact of counselling on exclusive breast-feeding practices in a poor urban setting in Kenya: a randomized controlled trial.
To determine the impact of facility-based semi-intensive and home-based intensive counselling in improving exclusive breast-feeding (EBF) in a low-resource urban setting in Kenya. A cluster randomized controlled trial in which nine villages were assigned on a 1:1:1 ratio, by computer, to two intervention groups and a control group. The home-based intensive counselling group (HBICG) received seven counselling sessions at home by trained peers, one prenatally and six postnatally. The facility-based semi-intensive counselling group (FBSICG) received only one counselling session prenatally. The control group (CG) received no counselling from the research team. Information on infant feeding practices was collected monthly for 6 months after delivery. The data-gathering team was blinded to the intervention allocation. The outcome was EBF prevalence at 6 months. Kibera slum, Nairobi. A total of 360 HIV-negative women, 34-36 weeks pregnant, were selected from an antenatal clinic in Kibera; 120 per study group. Of the 360 women enrolled, 265 completed the study and were included in the analysis (CG n 89; FBSICG n 87; HBICG n 89). Analysis was by intention to treat. The prevalence of EBF at 6 months was 23.6% in HBICG, 9.2% in FBSICG and 5.6% in CG. HBICG mothers had four times increased likelihood to practise EBF compared with those in the CG (adjusted relative risk = 4.01; 95% CI 2.30, 7.01; P=0.001). There was no significant difference between EBF rates in FBSICG and CG. EBF can be promoted in low socio-economic conditions using home-based intensive counselling. One session of facility-based counselling is not sufficient to sustain EBF.


A nonredundant role for the adapter protein Shc in thymic T cell development.
Signaling via the pre-T cell receptor (pre-TCR) regulates survival, proliferation, allelic exclusion and differentiation of thymocytes. The role played by the adapter protein Shc in T cells has remained controversial, and its role in pre-TCR signaling has not been addressed. We examined Shc function in thymic T cell development using two genetic approaches. Cre-loxP-mediated inducible expression in transgenic mice of a phosphorylation-defective mutant of Shc impaired signaling through the pre-TCR as well as subsequent proliferation and differentiation. Conditional deletion of the Shc locus in thymocytes also affected thymic maturation at the same pre-TCR developmental stage. Thus, both Shc expression and its tyrosine phosphorylation play an essential and nonredundant role in thymic T cell development.


Cardiovascular adaptation to mudpack therapy in hypertensive subjects treated with different antihypertensive drugs.
In selected hypertensive subjects, cardiovascular adaptation to warm environments may be inadequate or even harmful: heating associated to mudpack therapy may cause unexpected hypotension. How different antihypertensive drugs may affect the cardiovascular response to mudpack therapy is poorly studied. To evaluate the effects of β-blockers and angiotensin II receptor antagonists/ACE inhibitors on the acute cardiovascular adaptation to mudpack treatment in SPA in elderly hypertensive patients. Thirty-one elderly subjects were divided in normotensive subjects (N; n=10) and hypertensive patients treated with ACE-inhibitors/Angiotensin II receptor antagonists (HTA; n=12) or with selective β1-blockers (HTB; n=9). Systolic (SBP) and diastolic (DBP) blood pressure were continuously recorded (10 min) in supine position, immediately before and during mudpack treatment (40°C). Heart rate (HR), stroke volume (SV), cardiac output (CO) and total peripheral resistance (TPR) were assessed. During mudpack treatment SBP did not significantly change in both HTA and N groups (132±11 and 112±13 mmHg, respectively), but significantly decreased in HTB (111±18 mmHg, p < 0.01 vs baseline) patients. HR increased in all groups (HTA: 72±10 bpm; HTB: 65±6 bpm; N: 70±10 bpm; p < 0.01 vs baseline). A significant reduction (p < 0.01 vs baseline) in SV and CO occurred in HTB, but not in HTA and N groups. TPR significantly increased in HTB (1335±464 dyn.s.cm-5, p < 0.01 vs baseline) but not in HTA and N subjects (1389±385 dyn.s.cm-5 and 1245±323 dyn.s.cm-5, respectively). Mud treatment did not cause relevant haemodynamic changes in normotensive and HTA-treated hypertensive subjects. Conversely, β-blocking treatment apparently limited the cardiovascular adaptation to thermic stress, through a possible reduction in myocardial contractility, thereby, causing a significant decrease, although not dangerous, in systolic blood pressure.


Characteristics and Prognosis in Women and Men With Type 1 Diabetes Undergoing Coronary Angiography: A Nationwide Registry Report.
To describe sex aspects on extent of coronary artery disease (CAD) and prognosis in a contemporary population with type 1 diabetes. All patients undergoing coronary angiography, 2001-2013, included in the Swedish Coronary Angiography and Angioplasty Registry and the Swedish National Diabetes Register as type 1 diabetes were followed for mortality until 31 December 2013. The coronary angiogram was classified into normal, one-vessel disease, two-vessel disease, three-vessel disease, and left main stem disease. In all, 2,776 patients (42% women) with mean age 58 years (SD 11) were followed for 7.2 years (SD 2.2). Diabetes duration was longer in women (37 ± 14 vs. 34 ± 14 years in men; P < 0.001), who also had more retinopathy (68% vs. 65%; P = 0.050), whereas microalbuminuria was less common (41% vs. 51%; P < 0.001). Indications for coronary angiography did not substantially differ in women and men. The extent of CAD was somewhat less severe in women (normal angiogram 23.5% vs. 19.1%, three-vessel and left main stem disease 34.5% vs. 40.4%; P = 0.002), whereas mortality did not differ (adjusted hazard ratio 1.03 [95% CI 0.88-1.20]; P = 0.754). The standard mortality ratio for women the first year was 7.49 (5.73-9.62) and for men was 4.58 (3.60-5.74). In patients with type 1 diabetes admitted for coronary angiography, the extent of CAD was almost similar in women and men, and total long-term mortality did not differ. Type 1 diabetes was associated with higher mortality risk in women than in men when compared with the general population. These data support that type 1 diabetes attenuates the cardiovascular risk difference seen in men and women in the general population.


Clinical implications of plasma Nogo-A levels in patients with coronary heart disease.
Nogo-A is an important neurite growth-regulatory protein in the adult and developing nervous system. Recently, increasing evidence has shown that Nogo-A plays important roles in cardiac development and may act as a potential indicator for heart failure. In addition, increased oxidative stress has been found in individuals with cardiovascular diseases. However, not much is known regarding the expression levels of Nogo-A and reactive oxygen species (ROS) in patients with coronary heart disease (CHD). Therefore, we sought to investigate the relationship between Nogo-A, ROS levels and CHD. The plasma Nogo-A and ROS concentrations of 122 acute coronary syndrome (ACS), 101 unstable angina pectoris (UAP), and 21 acute myocardial infarction (AMI) patients and 56 healthy controls were measured by enzyme-linked immunosorbent assay (ELISA). We further generated a receiver operating characteristic (ROC) curve to assess the diagnostic accuracy of Nogo-A and ROS in CHD. The Nogo-A and ROS levels were significantly higher in patients with CHD than those in healthy controls. In addition, multivariate logistic regression analysis revealed that the level of Nogo-A (odds ratio (OR) = 1.624, 95% confidence interval: 1.125-2.293, p = 0.009) is a risk factor for prediction of CHD. Nogo-A has diagnostic value, with an optimal threshold of 5.466 ng/ml for maximized diagnostic performance (59% sensitivity and 78.6% specificity, area under curve, p < 0.05). However, ROS concentration is not a risk factor for prediction of CHD (OR = 0.999, 95% confidence interval: 0.997-1.001, p = 0.320). Increased plasma Nogo-A level may be associated with CHD.


Identification of QTL controlling meat quality traits in an F2 cross between two chicken lines selected for either low or high growth rate.
Meat technological traits (i.e. meat pH, water retention and color) are important considerations for improving further processing of chicken meat. These quality traits were originally characterized in experimental lines selected for high (HG) and low (LG) growth. Presently, quantitative trait loci (QTL) for these traits were analyzed in an F2 population issued from the HG x LG cross. A total of 698 animals in 50 full-sib families were genotyped for 108 microsatellite markers covering 21 linkage groups. The HG and LG birds exhibit large differences in body weight and abdominal fat content. Several meat quality traits [pH at 15 min post-slaughter (pH15) and ultimate pH (pHu), breast color-redness (BCo-R) and breast color-yellowness (BCo-Y)] were lower in HG chickens. In contrast, meat color-lightness (BCo-L) was higher in HG chickens, whereas meat drip loss (DL) was similar in both lines. HG birds were more active on the shackle line. Association analyses were performed using maximum-likelihood interval mapping in QTLMAP. Five genome-wide significant QTLs were revealed: two for pH15 on GGA1 and GGA2, one for DL on GGA1, one for BCo-R and one for BCo-Y both on GGA11. In addition, four suggestive QTLs were identified by QTLMAP for BCo-Y, pHu, pH15 and DL on GGA1, GGA4, GGA12 and GGA14, respectively. The QTL effects, averaged on heterozygous families, ranged from 12 to 31% of the phenotypic variance. Further analyses with QTLExpress confirmed the two genome-wide QTLs for meat color on GGA11, failed to identify the genome-wide QTL for pH15 on GGA2, and revealed only suggestive QTLs for pH15 and DL on GGA1. However, QTLExpress qualified the QTL for pHu on GGA4 as genome-wide. The present study identified genome-wide significant QTLs for all meat technological traits presently assessed in these chickens, except for meat lightness. This study highlights the effects of divergent selection for growth rate on some behavioral traits, muscle biochemistry and ultimately meat quality traits. Several QTL regions were identified that are worthy of further characterization. Some QTLs may in fact co-localize, suggesting pleiotropic effects for some chromosomal regions.


Control mechanism of spontaneous in vitro contractions of the opossum sphincter of Oddi.
We evaluated the control mechanism of peristaltic contractions in the opossum sphincter of Oddi (SO) by means of an in vitro preparation. At each of four sites spaced uniformly along the sphincter segment, a force transducer recorded contractions while a monopolar electrode recorded myoelectric activity. Spontaneous myoelectric and contractile activity occurred in 15 of the 20 intact SO specimens studied. Electrical recordings showed characteristic control waves and response activity. Each control wave was invariably accompanied by a phasic contraction, irrespective of whether or not response activity was superimposed on the control wave. The predominant motor activity of the SO was antegrade peristalsis. Retrograde peristalsis occurred when antegrade peristalsis failed to traverse the entire sphincter. Spontaneous SO phasic contractions were not antagonized by tetrodotoxin. Muscle rings sectioned from the SO exhibited spontaneous phasic contractions with a proximal-to-distal gradient of intrinsic contraction frequencies. We conclude that a) the frequency of SO phasic contractions is determined by control wave frequency, b) spontaneous SO peristalsis is myogenic in origin and may be modeled by a linear array of bidirectionally coupled relaxation oscillators, c) the predominance of antegrade peristalsis may be explained by a high-frequency oscillator in the proximal SO that drives the slower, more distal oscillators, d) retrograde peristalsis is initiated by an ectopic oscillator in the distal SO when antegrade contractions fail to propagate the entire length of the SO, and e) ectopic SO contractions can propagate retrograde when the more proximal oscillators are not in their absolutely refractory state.


Enhanced T cell responses to antigenic peptides targeted to B cell surface Ig, Ia, or class I molecules.
The helper T cell recognition of soluble globular protein antigens requires that the proteins be processed by an APC, releasing a peptide that is transported to and held on the APC surface where it is recognized by the specific T cell in conjunction with Ia. When cellular processing functions are blocked, APC lose their ability to present native antigens while retaining the capacity to activate T cells when provided with a cognate peptide fragment that contains the T cell antigenic determinant. In this report, we show that a peptide fragment of the soluble globular protein antigen tobacco hornworm moth cytochrome c, residues 92-103 containing an additional NH2-terminal cysteine residue (THMcCys92-103), is effectively presented by B cells to an I-Ek-restricted, THMc-specific T cell hybrid when covalently coupled to antibodies specific for B cell surface Ig, Ia (Ak), or class I (Kk). Maximal activation of the T cells to the THMcCys92-103-antibody conjugates is achieved with 1/100-1/1,000th of the peptide required using unconjugated THMcCys92-103 or THMcCys92-103 coupled to nonspecific antibody. The T cell response to the peptide antibody conjugates is MHC restricted, but unlike native cytochrome c-antibody conjugates, THMcCys92-103-antibody conjugates do not require processing and can be presented by paraformaldehyde-fixed B cells. The THMcCys92-103-antibody conjugate are nearly as effective when incubated with B cells, and the unbound conjugates washed away before addition of T cells as when continuously present in culture with T cells and B cells, indicating that the active peptide antibody conjugate is associated at the B cell surface. The presentation of THMcCys92-103 coupled to monovalent Fab fragments of rabbit anti-Ig antibodies is less effective than that of the peptide coupled to bivalent antibody when either live or fixed B cells are APC, indicating that the avidity for the APC surface afforded by bivalent binding may be important in the conjugate's antigenicity. The results presented here indicate that a T cell-antigenic peptide, covalently coupled to a larger antibody molecule, can be readily recognized by an Ia-restricted helper T cell in the absence of processing. Moreover, the ability of the peptide to bind to B cell surfaces greatly augments the peptide's antigenicity, even when the binding is to structures distinct from the Ia molecule required for T cell activation.


Modulation by progestogens of the effects of oestrogen on hepatic endocrine function in postmenopausal women.
Oral but not transdermal oestrogen administration reduces IGF-I, and increases GH binding protein (GHBP) reflecting effects on hepatic endocrine function in postmenopausal women. As progestogens attenuate the effects of oestrogen on circulating lipid levels according to their androgenic properties, we have investigated the impact of progestogen types on the hepatic endocrine effects of oestrogen. Four progestogens differing in androgenicity were co-administered in a monthly cyclical regimen in random order to postmenopausal women receiving either oral (n = 9, premarin 1.25 mg) or transdermal (n = 10, Estraderm 100 microg patches twice weekly). The four progestogens were cyproterone acetate (CA 5 mg, antiandrogenic), dydrogesterone (20 mg, neutral), medroxyprogesterone acetate (MPA 10 mg, mildly androgenic), norethisterone (2.5 mg, androgenic). Nineteen postmenopausal women (age 57 +/- 3 years, mean +/- SE) were studied. The effects of oestrogen alone and the combined effects with each progestogen type on IGF-I, GHBP, SHBG, cholesterol, triglycerides and lipoprotein(a) were investigated. Mean IGF-I fell while GHBP and SHBG levels increased with oral (P < 0.01) but not transdermal oestrogen administration. When the combined effects were examined, progestogens did not affect IGF-I, GHBP and SHBG during oral oestrogen treatment, while they significantly increased (P < 0.01) mean IGF-I levels during transdermal therapy. Among the progestogen types, only norethisterone prevented the fall in IGF-I induced by oral oestrogen. During transdermal therapy, MPA and norethisterone but not CA or dydrogesterone significantly increased (P < 0.005) IGF-I. The rise in GHBP induced by oral oestrogens tended to be lower during co-administration of MPA and norethisterone. The increase in SHBG induced by oral oestrogen was attenuated (P < 0.05) by norethisterone which was the only progestogen that lowered SHBG (P < 0.05) during transdermal oestrogen treatment. Mean IGF-I was higher (P < 0.001), GHBP and SHBG lower during co-administration of androgenic progestogens (MPA and norethisterone). Oestrogen effects on IGF-I, GHBP and SHBG are dependent on the route of administration with progestogens having variable effects. Among the progestogen types, norethisterone, the most androgenic, had the greatest effect, particularly on IGF-I. Progestogens modulate the effects of oestrogen on hepatic endocrine function in relation to their intrinsic androgenic properties. The modulatory effects of progestogens on IGF-I during oestrogen therapy may have long-term implications for lean body mass.


Diverse morphologies in thin-film bijels by varying film thickness and composition.
Thin-film bijels have many potential energy and environmental applications. In this work, the metastable bijel morphology space in thin-film confinement is explored with a Cahn-Hilliard/Brownian-Dynamics computational model. The key parameters varied are the bijel liquid phase blend ratio and the bijel film thickness. Simulations reveal a broad spectrum of structurally unique morphologies that have yet to be observed in experiments and which could have interesting applications in membrane science and other domains. Extensive analyses of surface-to-volume ratios, interfacial particle attachment statistics, and topological interfacial curvatures within the bijels are presented for a complete characterization of the morphological structure.


Differences in strength and power among junior-high, senior-high, college-aged, and elite professional rugby league players.
Ninety-five rugby league players ranging from junior high-school to elite professionals were compared for measures of strength and power. Strength was assessed by 1 repetition maximum bench press strength (1RM BP). Upper-body and lower-body power outputs were assessed during bench press throws and jump squats with a resistance of 20 kg (BT P20 and JS P20, respectively). The 1RM BP was a potent descriptor of playing achievement levels and was significantly different among all groups investigated. Both the BT P20 and JS P20 of the elite professional National Rugby League (NRL) group were significantly higher than those of the college-aged rugby league (CRL) group, which in turn were significantly higher than those of the 3 high-school groups. Senior high-school players were more powerful in the upper body compared with nonresistance-trained junior high-school players but not with resistance-trained junior high-school players. There was no difference in lower-body power output among any of the 3 high-school groups. The correlation between players achievement level and 1RM BP, BT P20, and JS P20 was significant for all 3 tests, with relations of r = 0.80, r = 0.74, and r = 0.61, respectively. The results of this study suggest that young rugby league players should strive to increase strength and power to attain NRL professional status in the future.


Targeted focal therapy: a minimally invasive ablation technique for early prostate cancer.
The morbidities associated with prostate cancer treatments have improved over the years. However, potential overtreatment and the risks of adverse events associated with radical treatment still pose a considerable challenge. Targeted focal therapy (TFT) of prostate cancer appears to be part of a logical continuum in the quest to improve upon the management of early organ-confined disease. TFT is a procedure in which only the cancer in the gland is ablated. The normal gland, sphincter, and in most cases the neurovascular bundles are preserved. Therefore, this approach averts some of the common complications of more radical therapy. Initial experience has been encouraging; however, long-term data and full implementation of emerging advances in imaging are urgently needed before the widespread adoption of this approach. In this review, we present the current status of our knowledge about this procedure and the most important challenges that need to be addressed. We also present the initial results with this approach at our center.


Proceedings of the 25th Analytical Ultracentrifugation Workshops and Symposium.
The 25th International Analytical Ultracentrifugation (AUC) Workshops and Symposium (AUC2022) took place at the University of Lethbridge in Lethbridge, Canada, in July 2022. In total, 104 attendees (Attendance Profile: 104 attendees, 69 in-person, 35 remote. Brazil 1, Canada 24, China 1, Czech Republic 2, Finland 1, France 3, Germany 22, India 3, Italy 1, Japan 4, Spain 1, Switzerland 3, Taiwan 1, United Kingdom 5, United States 32) participated in the event and presented the latest advances in the field. While the primary focus of the conference was to showcase the applications of AUC in chemical, life sciences, and nanoparticle disciplines, several presentations also integrated complementary methods, such as isothermal titration calorimetry, microscale thermophoresis, light scattering (static and dynamic), small-angle X-ray scattering, X-ray crystallography, and cryo-electron microscopy. Additionally, the delegates gained valuable hands-on experience from 20 workshops covering a broad range of applications, experimental designs and systems, and the latest software innovations in solution biophysics. The AUC2022 special volume highlights the sustained innovation, utility and relevance of AUC and related solution biophysical methods across various disciplines, including biochemistry, structural biology, synthetic polymer chemistry, carbohydrate chemistry, protein and nucleic acid characterization, nano-science, and macromolecular interactions.


Bone Marrow Adipose Tissue: A New Player in Cancer Metastasis to Bone.
The bone marrow is a favored site for a number of cancers, including the hematological malignancy multiple myeloma, and metastasis of breast and prostate cancer. This specialized microenvironment is highly supportive, not only for tumor growth and survival but also for the development of an associated destructive cancer-induced bone disease. The interactions between tumor cells, osteoclasts and osteoblasts are well documented. By contrast, despite occupying a significant proportion of the bone marrow, the importance of bone marrow adipose tissue is only just emerging. The ability of bone marrow adipocytes to regulate skeletal biology and hematopoiesis, combined with their metabolic activity, endocrine functions, and proximity to tumor cells means that they are ideally placed to impact both tumor growth and bone disease. This review discusses the recent advances in our understanding of how marrow adipose tissue contributes to bone metastasis and cancer-induced bone disease.


Two-hundredfold volume concentration of dilute cell and particle suspensions using chip integrated multistage acoustophoresis.
Concentrating cells is a frequently performed step in cell biological assays and medical diagnostics. The commonly used centrifuge exhibits limitations when dealing with rare cell events and small sample volumes. Here, we present an acoustophoresis microfluidic chip utilising ultrasound to concentrate particles and cells into a smaller volume. The method is label-free, continuous and independent of suspending fluid, allowing for low cost and minimal preparation of the samples. Sequential concentration regions and two-dimensional acoustic standing wave focusing of cells and particles were found critical to accomplish concentration factors beyond one hundred times. Microparticles (5 μm in diameter) used to characterize the system were concentrated up to 194.2 ± 9.6 times with a recovery of 97.1 ± 4.8%. Red blood cells and prostate cancer cells were concentrated 145.0 ± 5.0 times and 195.7 ± 36.2 times, respectively, with recoveries of 97.2 ± 3.3% and 97.9 ± 18.1%. The data demonstrate that acoustophoresis is an effective technique for continuous flow-based concentration of cells and particles, offering a much needed intermediate step between sorting and detection of rare cell samples in lab-on-a-chip systems.


Protein losing enteropathy associated with Nippostrongylus brasiliensis infestation and its impact on albumin homoeostasis in rats fed two levels of dietary protein.
Alterations in plasma albumin concentration and gastrointestinal permeability have been investigated in rats infected with the nematode Nippostrongylus brasiliensis and fed adequate or low protein diets. Infection caused only minor changes in growth and food consumption of well nourished rats but resulted in significant reductions in those fed the low protein diet. Animals in both dietary groups were able to mount an immune response beyond day 10 post-infection (p.i.) which caused expulsion of the parasites, but this was less effective in rats fed the low protein food. Uninfected rats fed the low protein diet had significantly lower plasma albumin concentrations than their well nourished counterparts. Animals of both dietary groups showed a progressive reduction in plasma albumin concentration as the infection developed but values returned towards normal as the parasites were expelled. The reduction in plasma albumin concentration was closely associated with increases in gastrointestinal leakage of plasma protein but losses were far greater in the protein deficiency animals. Beyond day 10 p.i. protein loss decreased in both dietary groups and by day 21 p.i. had returned to normal in well nourished animals but not those fed the low protein diet. Intestinal permeability measured by the lactulose:mannitol ratio technique gave similar results to the protein loss data. Permeability increased as the infection progressed then fell as the worms were expelled but remained above control values in infected protein deficient animals. Overall, animals fed the low protein diet were more severely affected by the parasite than were their well fed counterparts.(ABSTRACT TRUNCATED AT 250 WORDS)


Platelet calcium transport in hypertension.
To determine platelet Ca2+ transport entities involved in increased cytosolic Ca2+ in the platelets of hypertensive individuals, we studied the relations between blood pressure and Ca2+ transporters in platelet membranes from 22 white male volunteers 32 to 68 years old. We used thapsigargin, a specific inhibitor of the internal membrane Ca(2+)-ATPase, to differentiate between plasma membrane and internal membrane Ca(2+)-ATPases. Inositol 1,4,5-trisphosphate-mediated and Ca2+ ionophore (A23187)-induced Ca2+ release was also assayed in membrane preparations using rhod-2, a fluorescent Ca2+ indicator. Levels of glycoprotein IIIa, a possible component of agonist-mediated Ca2+ influx, were measured by immunoblotting. The results show that plasma membrane Ca(2+)-ATPase is decreased as a function of diastolic blood pressure (P < .002), whereas the internal membrane Ca(2+)-ATPase is not (P < .148). Neither activity is correlated with age or systolic blood pressure. However, inositol trisphosphate-mediated Ca2+ release is negatively correlated with age (P < .024) but not blood pressure. Glycoprotein IIIa levels and A23187-induced Ca2+ release were not related to age or blood pressure, demonstrating that inhibition of the plasma membrane Ca(2+)-ATPase was not a result of differences in the proportion of plasma membrane in the preparation or differences in intravesicular Ca2+ concentration. Inhibition of the plasma membrane Ca(2+)-ATPase could directly cause elevation of cytoplasmic Ca2+ and enhancement of platelet sensitivity.


Management of multiple myeloma in the relapsed/refractory patient.
The approach to the patient with relapsed or relapsed/refractory multiple myeloma requires a careful evaluation of the results of previous treatments, the toxicities associated with it, and an assessment of prognostic factors. The majority of patients will have received prior therapy with drug combinations, including a proteasome inhibitor and an immune-modulatory agent. It is the physician's task to choose the right moment for the start of therapy and decide with the patient which goals need to be achieved. The choice of regimen is usually based on prior response, drugs already received, adverse effects, comorbidities of the patient, and expected efficacy and tolerability. Many double and triple drug combinations are available. In addition, promising new drugs such as pomalidomide, carfilzomib, and monoclonal antibodies are or will be available shortly, and other options can be explored in clinical trials. Finally, supportive care and palliative options need to be considered in later relapsed disease. Increasingly, it becomes important to consider the therapeutic options for the whole duration of the disease and integrate a systematic approach for the patient.


Closed-Loop Insulin Delivery during Pregnancy in Women with Type 1 Diabetes.
In patients with type 1 diabetes who are not pregnant, closed-loop (automated) insulin delivery can provide better glycemic control than sensor-augmented pump therapy, but data are lacking on the efficacy, safety, and feasibility of closed-loop therapy during pregnancy. We performed an open-label, randomized, crossover study comparing overnight closed-loop therapy with sensor-augmented pump therapy, followed by a continuation phase in which the closed-loop system was used day and night. Sixteen pregnant women with type 1 diabetes completed 4 weeks of closed-loop pump therapy (intervention) and sensor-augmented pump therapy (control) in random order. During the continuation phase, 14 of the participants used the closed-loop system day and night until delivery. The primary outcome was the percentage of time that overnight glucose levels were within the target range (63 to 140 mg per deciliter [3.5 to 7.8 mmol per liter]). The percentage of time that overnight glucose levels were in the target range was higher during closed-loop therapy than during control therapy (74.7% vs. 59.5%; absolute difference, 15.2 percentage points; 95% confidence interval, 6.1 to 24.2; P=0.002). The overnight mean glucose level was lower during closed-loop therapy than during control therapy (119 vs. 133 mg per deciliter [6.6 vs. 7.4 mmol per liter], P=0.009). There were no significant differences between closed-loop and control therapy in the percentage of time in which glucose levels were below the target range (1.3% and 1.9%, respectively; P=0.28), in insulin doses, or in adverse-event rates. During the continuation phase (up to 14.6 additional weeks, including antenatal hospitalizations, labor, and delivery), glucose levels were in the target range 68.7% of the time; the mean glucose level was 126 mg per deciliter (7.0 mmol per liter). No episodes of severe hypoglycemia requiring third-party assistance occurred during either phase. Overnight closed-loop therapy resulted in better glucose control than sensor-augmented pump therapy in pregnant women with type 1 diabetes. Women receiving day-and-night closed-loop therapy maintained glycemic control during a high proportion of the time in a period that encompassed antenatal hospital admission, labor, and delivery. (Funded by the National Institute for Health Research and others; Current Controlled Trials number, ISRCTN71510001.).


Association of Obsessive-Compulsive Disorder With Objective Indicators of Educational Attainment: A Nationwide Register-Based Sibling Control Study.
To our knowledge, the association of obsessive-compulsive disorder (OCD) and academic performance has not been objectively quantified. To investigate the association of OCD with objectively measured educational outcomes in a nationwide cohort, adjusting for covariates and unmeasured factors shared between siblings. This population-based birth cohort study included 2 115 554 individuals who were born in Sweden between January 1, 1976, and December 31, 1998, and followed up through December 31, 2013. Using the Swedish National Patient Register and previously validated International Statistical Classification of Diseases and Related Health Problems, Tenth Revision (ICD-10) codes, we identified persons with OCD; within the cohort, we identified 726 198 families with 2 or more full siblings, and identified 11 482 families with full siblings discordant for OCD. Data analyses were conducted from October 1, 2016, to September 25, 2017. The study evaluates the following educational milestones: eligibility to access upper secondary school after compulsory education, finishing upper secondary school, starting a university degree, finishing a university degree, and finishing postgraduate education. Of the 2 115 554 individuals in the cohort, 15 120 were diagnosed with OCD (59% females). Compared with unexposed individuals, those with OCD were significantly less likely to pass all core and additional courses at the end of compulsory school (adjusted odds ratio [aOR] range, 0.35-0.60) and to access a vocational or academic program in upper secondary education (aOR, 0.47; 95% CI, 0.45-0.50 and aOR, 0.61; 95% CI, 0.58-0.63, for vocational and academic programs, respectively). People with OCD were also less likely to finish upper secondary education (aOR, 0.43; 95% CI, 0.41-0.44), start a university degree (aOR, 0.72; 95% CI, 0.69-0.75), finish a university degree (aOR, 0.59; 95% CI, 0.56-0.62), and finish postgraduate education (aOR, 0.52; 95% CI, 0.36-0.77). The results were similar in the sibling comparison models. Individuals diagnosed with OCD before age 18 years showed worse educational attainment across all educational levels compared with those diagnosed at or after age 18 years. Exclusion of patients with comorbid neuropsychiatric disorders, psychotic, anxiety, mood, substance use, and other psychiatric disorders resulted in attenuated estimates, but patients with OCD were still impaired across all educational outcomes. Obsessive-compulsive disorder, particularly when it has an early onset, is associated with a pervasive and profound decrease in educational attainment, spanning from compulsory school to postgraduate education.


Inhibition of Long-Term Variability in Decoding Forelimb Trajectory Using Evolutionary Neural Networks With Error-Correction Learning.
Objective: In brain machine interfaces (BMIs), the functional mapping between neural activities and kinematic parameters varied over time owing to changes in neural recording conditions. The variability in neural recording conditions might result in unstable long-term decoding performance. Relevant studies trained decoders with several days of training data to make them inherently robust to changes in neural recording conditions. However, these decoders might not be robust to changes in neural recording conditions when only a few days of training data are available. In time-series prediction and feedback control system, an error feedback was commonly adopted to reduce the effects of model uncertainty. This motivated us to introduce an error feedback to a neural decoder for dealing with the variability in neural recording conditions. Approach: We proposed an evolutionary constructive and pruning neural network with error feedback (ECPNN-EF) as a neural decoder. The ECPNN-EF with partially connected topology decoded the instantaneous firing rates of each sorted unit into forelimb movement of a rat. Furthermore, an error feedback was adopted as an additional input to provide kinematic information and thus compensate for changes in functional mapping. The proposed neural decoder was trained on data collected from a water reward-related lever-pressing task for a rat. The first 2 days of data were used to train the decoder, and the subsequent 10 days of data were used to test the decoder. Main Results: The ECPNN-EF under different settings was evaluated to better understand the impact of the error feedback and partially connected topology. The experimental results demonstrated that the ECPNN-EF achieved significantly higher daily decoding performance with smaller daily variability when using the error feedback and partially connected topology. Significance: These results suggested that the ECPNN-EF with partially connected topology could cope with both within- and across-day changes in neural recording conditions. The error feedback in the ECPNN-EF compensated for decreases in decoding performance when neural recording conditions changed. This mechanism made the ECPNN-EF robust against changes in functional mappings and thus improved the long-term decoding stability when only a few days of training data were available.


Effect of thioacetamide on rat liver regeneration. II. Nuclear RNA in mitosis.
The effect of thioacetamide on dividing cells of regenerating rat liver has been studied. Rats were given daily subcutaneous injections of thioacetamide as a 1 per cent solution at a dosage of 5 mg./100 gm. body weight for 7 to 10 days, subjected to partial hepatectomy, and sacrificed 28 to 31 hours later. Thioacetamide treatment results in striking increases in the nuclear ribonucleoproteins of the liver cell without affecting the mitotic rate during regeneration (14). During mitosis, RNA-containing particles were seen within the spindle and coating the contracted chromosomes from prophase through metaphase or early anaphase. At telophase, prior to the reconstruction of the nuclear membrane, fine RNA-containing granules appeared within the compact chromosomal groups. These coalesced to form nucleoli corresponding in number to the number of nucleolar organizer regions. The nuclei and nucleoli showed a rapid increase in size during the reconstruction period when compared with corresponding figures of the control liver samples. Electron micrographs of interphase nucleoli indicated a similar basic granular structure in both drug-treated and control animals. The question is raised as to whether the increased nucleolar material merely made visible some of the nucleolar-chromosomal associations that normally occur in mitosis, or whether thioacetamide directly affects the synthetic activity of the contracted mitotic chromosomes.


Recombinant human growth hormone in patients with HIV-associated wasting. A randomized, placebo-controlled trial. Serostim Study Group.
Body wasting, particularly loss of body cell mass, is an increasingly prevalent acquired immunodeficiency syndrome (AIDS)-defining condition and is an independent risk factor for death in patients infected with the human immunodeficiency virus (HIV). Treatment with growth hormone for 7 days resulted in weight gain and nitrogen retention, but the long-term effects of this treatment in patients with HIV-associated wasting are not known. To evaluate the long-term effect of treatment with growth hormone on weight, body composition, functional performance, and quality of life in patients with HIV-associated wasting. Randomized, double-blind, placebo-controlled, multicenter trial. Outpatient university and community-based patient care facilities. 178 HIV-infected patients with documented unintentional weight loss of at least 10% or weight less than 90% of the lower limit of ideal body weight. Patients were randomly assigned to receive either recombinant human growth hormone, 0.1 mg/kg of body weight per day (average dosage, 6 mg/d) (n = 90) or placebo (n = 88) for 12 weeks. Weight; body fat, lean body mass, and bone mineral content (measured by dual-energy x-ray absorptiometry); total body water (by deuterium oxide dilution); extracellular water (by sodium bromide dilution); work output (by treadmill exercise); quality of life; and safety of treatment. Treatment with growth hormone resulted in a sustained and statistically significant increase in weight (mean increase +/- SD, 1.6 +/- 3.7 kg [P < 0.001]) and lean body mass (3.0 +/- 3.0 kg [P < 0.001]), accompanied by a decrease in body fat (-1.7 +/- 1.7 kg [P < 0.001]). In contrast, in patients receiving placebo, weight (increase, 0.1 +/- 3.1 kg), lean body mass (decrease, 0.1 +/- 2.0 kg), and body fat (decrease, 0.3 +/- 2.2 kg) did not change significantly from baseline. Differences between groups at week 12 were statistically significant (P = 0.011 for body weight and P < 0.001 for lean body mass and body fat). A greater increase in treadmill work output was noted in the group receiving growth hormone (increase, 99 +/- 293 kg. m/min) compared with the group receiving placebo (increase, 20 +/- 233 kg.m/min)(P = 0.039). Health status (quality of life) scores did not differ between groups at baseline or after treatment. Days of disability and use of medical resources were the same for both groups. Treatment was was well tolerated; no significant differences were seen between groups in clinical events, progression of AIDS, CD4+ or CD8+ cell counts, or viral burden. Treatment with growth hormone increases body weight, lean body mass, and treadmill work output and appears to be a safe and potentially effective therapy in patients with HIV-associated wasting.


Final version of 2009 AJCC melanoma staging and classification.
To revise the staging system for cutaneous melanoma on the basis of data from an expanded American Joint Committee on Cancer (AJCC) Melanoma Staging Database. The melanoma staging recommendations were made on the basis of a multivariate analysis of 30,946 patients with stages I, II, and III melanoma and 7,972 patients with stage IV melanoma to revise and clarify TNM classifications and stage grouping criteria. Findings and new definitions include the following: (1) in patients with localized melanoma, tumor thickness, mitotic rate (histologically defined as mitoses/mm(2)), and ulceration were the most dominant prognostic factors. (2) Mitotic rate replaces level of invasion as a primary criterion for defining T1b melanomas. (3) Among the 3,307 patients with regional metastases, components that defined the N category were the number of metastatic nodes, tumor burden, and ulceration of the primary melanoma. (4) For staging purposes, all patients with microscopic nodal metastases, regardless of extent of tumor burden, are classified as stage III. Micrometastases detected by immunohistochemistry are specifically included. (5) On the basis of a multivariate analysis of patients with distant metastases, the two dominant components in defining the M category continue to be the site of distant metastases (nonvisceral v lung v all other visceral metastatic sites) and an elevated serum lactate dehydrogenase level. Using an evidence-based approach, revisions to the AJCC melanoma staging system have been made that reflect our improved understanding of this disease. These revisions will be formally incorporated into the seventh edition (2009) of the AJCC Cancer Staging Manual and implemented by early 2010.


Phase II trial on the effects of Silexan in patients with neurasthenia, post-traumatic stress disorder or somatization disorder.
Silexan, a novel lavender oil preparation for oral use, has been authorized in Germany for the treatment of states of restlessness during anxious mood. An open-label, exploratory trial was performed to assess the potential of the medicinal product in the treatment of restlessness caused by anxiety as related to several disorders. Outcome measures included the Symptom Checklist-90-Revised (SCL-90-R), von Zerssen's Depression Scale (D-S), the 36-item Short Form Health Survey Questionnaire (SF-36), and a sleep diary. 50 male and female patients with neurasthenia (ICD-10 F48.0), post-traumatic stress disorder (PSD; F43.1), or somatization disorder (F45.0, F45.1) were included to receive 1 × 80 mg/day Silexan over 6 weeks; 47 could be analyzed for efficacy as full analysis set. At baseline, patients suffered from restlessness (96%), depressed mood (98%), sleep disturbances (92%), or anxiety (72%). Of those, resp. 62%, resp. 57%, resp.51%, resp. 62% showed improvements during treatment (p < 0.001). For all patients, mean D-S score decreased by 32.7% and SCL-90-R Global Severity Index by 36.4% as compared to baseline, (p < 0.001), while the SF-36 Mental Health Score increased by 48.2% (p < 0.001). Waking-up frequency (p = 0.002), Waking-up duration (p < 0.001) and morning tiredness (p = 0.005) were reduced, while efficiency of sleep (p = 0.018) and mood (p = 0.03) improved. Patients suffering from neurasthenia or PSD showed comparable improvements with most outcomes. The results in this trial justify to further investigate Silexan in disorders with accompanying restlessness caused by sub-threshold anxiety. Adverse reactions, predominantly gastrointestinal complaints, were judged as mild or moderate.


Menstrual and reproductive factors and gastric cancer risk in a large prospective study of women.
Gastric cancer incidence rates are consistently lower in women than men in both high and low-risk regions worldwide. Sex hormones, such as progesterone and estrogen, may protect women against gastric cancer. To investigate the association of menstrual and reproductive factors and gastric cancer risk. These associations were prospectively investigated in 73 442 Shanghai women. After 419 260 person-years of follow-up, 154 women were diagnosed with gastric cancer. Hazard ratios (HR) and 95% confidence intervals (CI) were calculated using Cox proportional hazards models adjusted for age, body mass index, education, income, and cigarette use. No associations were observed between gastric cancer risk and age of menarche, number of children, breast feeding, or oral contraceptive use. In contrast, associations were observed with age of menopause (HR 0.80 per five-year increase in menopausal age, 95% CI 0.66-0.97), years of fertility (participants with less than 30 years of fertility were at increased risk compared with those with 30-36 years of fertility, HR 1.90, 95% CI 1.25-2.90), years since menopause (HR 1.26 per five years, 95% CI 1.03-1.53), and intrauterine device use (HR for users 1.61, 95% CI 1.08-2.39). These results support the hypothesis that female hormones play a protective role in gastric cancer risk.


Clinical outcomes and prognostic factors of transcatheter aortic valve implantation in bicuspid aortic valve patients.
The purpose of this study was to evaluate the outcomes of transcatheter aortic valve replacement (TAVR) in patients with bicuspid aortic valve stenosis (AS). From April 2012 and December 2016, 108 patients with bicuspid AS underwent TAVR using the Sapien XT (34 patients) and Sapien 3 (74 patients) valves. Procedural and clinical outcomes were assessed according to VARC-2 criteria and compared between the two devices. In the overall cohort, the majority of patients were male (71.3%) with an intermediate surgical risk and a mean Society of Thoracic Surgeons (STS) score of 5.2%. Compared to the Sapien XT group, the Sapien 3 group had a significantly lower STS score (3.3%±2.0% vs. 6.7%±3.6%; P=0.001). Compared to the Sapien XT group, the Sapien 3 group had a significantly lower rate of moderate or severe paravalvular leak (2.7% vs. 14.7%; P=0.03) and higher device success (97.3% vs. 82.4%; P=0.006). There were no significant differences between the two groups in terms of 30-day all-cause mortality, stroke, life-threatening bleeding, major vascular complication and acute kidney injury (stage 2 or 3). Cumulative all-cause mortality at 1-year follow-up was 6.9%. There were no significant differences in cumulative event rates for all-cause mortality at 1-year follow-up between the two groups (9.4% vs. 4.6%; log-rank P=0.47). By univariate analysis, major vascular complication was significantly associated with overall all-cause mortality [hazard ratios (HR): 7.57; 95% confidence interval (CI): 1.51-37.86; P=0.014]. TAVR using the balloon-expandable valves provided acceptable procedural and clinical outcomes in patients with bicuspid AS. The new-generation Sapien 3 valves showed improved procedural outcomes compared to the early-generation Sapien XT valves.


The 46/50 kDa phosphoprotein VASP purified from human platelets is a novel protein associated with actin filaments and focal contacts.
Vasoactive agents which elevate either cGMP or cAMP inhibit platelet activation by pathways sharing at least one component, the 46/50 kDa vasodilator-stimulated phosphoprotein (VASP). VASP is stoichiometrically phosphorylated by both cGMP-dependent and cAMP-dependent protein kinases in intact human platelets, and its phosphorylation correlates very well with platelet inhibition caused by cGMP- and cAMP-elevating agents. Here we report that in human platelets spread on glass, VASP is associated predominantly with the distal parts of radial microfilament bundles and with microfilaments outlining the periphery, whereas less VASP is associated with a central microfilamentous ring. VASP is also detectable in a variety of different cell types including fibroblasts and epithelial cells. In fibroblasts, VASP is concentrated at focal contact areas, along microfilament bundles (stress fibres) in a punctate pattern, in the periphery of protruding lamellae, and is phosphorylated by cGMP- and cAMP-dependent protein kinases in response to appropriate stimuli. Evidence for the direct binding of VASP to F-actin is also presented. The data demonstrate that VASP is a novel phosphoprotein associated with actin filaments and focal contact areas, i.e. transmembrane junctions between microfilaments and the extracellular matrix.


The type VI secretion system of Vibrio cholerae fosters horizontal gene transfer.
Natural competence for transformation is a common mode of horizontal gene transfer and contributes to bacterial evolution. Transformation occurs through the uptake of external DNA and its integration into the genome. Here we show that the type VI secretion system (T6SS), which serves as a predatory killing device, is part of the competence regulon in the naturally transformable pathogen Vibrio cholerae. The T6SS-encoding gene cluster is under the positive control of the competence regulators TfoX and QstR and is induced by growth on chitinous surfaces. Live-cell imaging revealed that deliberate killing of nonimmune cells via competence-mediated induction of T6SS releases DNA and makes it accessible for horizontal gene transfer in V. cholerae.


Gene Expression Analysis Reveals Novel Shared Gene Signatures and Candidate Molecular Mechanisms between Pemphigus and Systemic Lupus Erythematosus in CD4+ T Cells.
Pemphigus and systemic lupus erythematosus (SLE) are severe potentially life-threatening autoimmune diseases. They are classified as B-cell-mediated autoimmune diseases, both depending on autoreactive CD4+ T lymphocytes to modulate the autoimmune B-cell response. Despite the reported association of pemphigus and SLE, the molecular mechanisms underlying their comorbidity remain unknown. Weighted gene co-expression network analysis (WGCNA) of publicly available microarray datasets of CD4+ T cells was performed, to identify shared gene expression signatures and putative overlapping biological molecular mechanisms between pemphigus and SLE. Using WGCNA, we identified 3,280 genes co-expressed genes and 14 co-expressed gene clusters, from which one was significantly upregulated for both diseases. The pathways associated with this module include type-1 interferon gamma and defense response to viruses. Network-based meta-analysis identified RSAD2 to be the most highly ranked hub gene. By associating the modular genes with genome-wide association studies (GWASs) for pemphigus and SLE, we characterized IRF8 and STAT1 as key regulatory genes. Collectively, in this in silico study, we identify novel candidate genetic markers and pathways in CD4+ T cells that are shared between pemphigus and SLE, which in turn may facilitate the identification of novel therapeutic targets in these diseases.


The transcription factors Scl and Lmo2 act together during development of the hemangioblast in zebrafish.
The transcription factors Scl and Lmo2 are crucial for development of all blood. An important early requirement for Scl in endothelial development has also been revealed recently in zebrafish embryos, supporting previous findings in scl(-/-) embryoid bodies. Scl depletion culminates most notably in failure of dorsal aorta formation, potentially revealing a role in the formation of hemogenic endothelium. We now present evidence that the requirements for Lmo2 in zebrafish embryos are essentially the same as for Scl. The expression of important hematopoietic regulators is lost, reduced, or delayed, panendothelial gene expression is down-regulated, and aorta-specific marker expression is lost. The close similarity of the phenotypes for Scl and Lmo2 suggest that they perform these early functions in hemangioblast development within a multiprotein complex, as shown for erythropoiesis. Consistent with this, we find that scl morphants cannot be rescued by a non-Lmo2-binding form of Scl but can be rescued by non-DNA-binding forms, suggesting tethering to target genes through DNA-binding partners linked via Lmo2. Interestingly, unlike other hematopoietic regulators, the Scl/Lmo2 complex does not appear to autoregulate, as neither gene's expression is affected by depletion of the other. Thus, expression of these critical regulators is dependent on continued expression of upstream regulators, which may include cell-extrinsic signals.


Oral specific desensitization in food-allergic children.
The possibility of obtaining oral desensitization in patients with food allergy is still a matter of debate. We decided to evaluate the safety and efficacy of standardized protocols for oral desensitization with the most common food allergens. Forty-two children (ages up to 16 years) diagnosed as affected by food allergy (on the basis of clinical history, skin prick tests, measurement of specific IgE, and double-blind, placebo-controlled food challenge) underwent a sublingual-oral desensitizing treatment according to new standardized protocols. The control group consisted of 10 patients who followed an elimination diet. The treatment was successfully completed by 85.7% of the patients. Specific IgE showed a significant decrease, while specific IgG(4) showed a significant increase, in all treated patients. The immunological modifications observed in our patients lead us to hypothesize that oral tolerance may be mediated by the same mechanisms as those involved in traditional desensitizing treatments for respiratory and insect sting allergy.


On the in vivo systolic compressibility of left ventricular free wall myocardium in the normal and infarcted heart.
Although studied for many years, there remain continued gaps in our fundamental understanding of cardiac kinematics, such as the nature and extent of heart wall volumetric changes that occur over the cardiac cycle. Such knowledge is especially important for accurate in silico simulations of cardiac pathologies and in the development of novel therapies for their treatment. A prime example is myocardial infarction (MI), which induces profound, regionally variant maladaptive remodeling of the left ventricle (LV) wall. To address this problem, we conducted an in vivo fiduciary marker-based study in an established ovine model of MI to generate detailed, time-evolving transmural in vivo volumetric measurements of LV free wall deformations in the normal state, as well as up to 12 h post-MI. This was accomplished using a transmural array of sonomicrometry crystals that acquired fiducial positions at ∼250 Hz with a positional accuracy of ∼0.1 mm, covering the entire infarct, border, and remote zones. A convex-hull method was used to directly calculate the Jacobian J(t)=Δv(t)/ΔVED from sonocrystal positions over the entire cardiac cycle, where ΔV is the volume of each convex polyhedral at end diastole (ED) (typically ∼1 cc). We demonstrated significant in vivo compressibility in normal functioning LV free wall myocardium, with JES=0.85±0.07 at end systole (ES). We also observed substantial regional variations, with the largest reduction in local myocardial tissue volume during systole in the base region accompanied by substantial transmural gradients. These patterns changed profoundly following loss of perfusion post-MI, with the apical region showing the greatest loss of volume reduction at ES. To verify that the sonocrystals did not affect local volumetric measurements, JES measures were also verified by non-invasive magnetic resonance imaging, exhibiting very similar changes in regional volume. We note that while our estimates of regional compressibility were in close agreement with the values previously reported for large animals, ranging from 5% to 20%, the direct, comprehensive measurements of wall compressibility presented herein improved on the limitations of previous reports. These limitations included dependency on the small local volumes used for analysis and often indirect measurement of compressibility. Our novel findings suggest that proper accounting for the myocardial effective compressibility at the ∼1 cc volume scale can improve the accuracy of existing kinematic indices, such as wall thickening and axial shortening, and simulations of LV remodeling following MI.