Resistance to ROS1 inhibition mediated by EGFR pathway activation in non-small cell lung cancer.
The targeting of oncogenic 'driver' kinases with small molecule inhibitors has proven to be a highly effective therapeutic strategy in selected non-small cell lung cancer (NSCLC) patients. However, acquired resistance to targeted therapies invariably arises and is a major limitation to patient care. ROS1 fusion proteins are a recently described class of oncogenic driver, and NSCLC patients that express these fusions generally respond well to ROS1-targeted therapy. In this study, we sought to determine mechanisms of acquired resistance to ROS1 inhibition. To accomplish this, we analyzed tumor samples from a patient who initially responded to the ROS1 inhibitor crizotinib but eventually developed acquired resistance. In addition, we generated a ROS1 inhibition-resistant derivative of the initially sensitive NSCLC cell line HCC78. Previously described mechanisms of acquired resistance to tyrosine kinase inhibitors including target kinase-domain mutation, target copy number gain, epithelial-mesenchymal transition, and conversion to small cell lung cancer histology were found to not underlie resistance in the patient sample or resistant cell line. However, we did observe a switch in the control of growth and survival signaling pathways from ROS1 to EGFR in the resistant cell line. As a result of this switch, ROS1 inhibition-resistant HCC78 cells became sensitive to EGFR inhibition, an effect that was enhanced by co-treatment with a ROS1 inhibitor. Our results suggest that co-inhibition of ROS1 and EGFR may be an effective strategy to combat resistance to targeted therapy in some ROS1 fusion-positive NSCLC patients.


Slow dissolving of emotional distress contributes to hyperarousal.
The mechanisms underlying hyperarousal, the key symptom of insomnia, have remained elusive, hampering cause-targeted treatment. Recently, restless rapid-eye-movement (REM) sleep emerged as a robust signature of sleep in insomnia. Given the role of REM sleep in emotion regulation, we hypothesized that restless REM sleep could interfere with the overnight resolution of emotional distress, thus contributing to accumulation of arousal. Participants (n = 1,199) completed questionnaires on insomnia severity, hyperarousal, self-conscious emotional distress, and thought-like nocturnal mentation that was validated to be a specific proxy for restless REM sleep (selective fragmentation: R = 0.57, P < 0.001; eye movement density: R = 0.46, P < 0.01) in 32 polysomnographically assessed participants. The experience of distress lasting overnight increased with insomnia severity (β = 0.29, P < 10(-23)), whereas short-lasting distress did not (β = -0.02, P = 0.41). Insomnia severity was associated with hyperarousal (β = 0.47, P < 10(-63)) and with the thought-like nocturnal mentation that is specifically associated with restless REM sleep (β = 0.31, P < 10(-26)). Structural equation modeling showed that 62.4% of the association between these key characteristics of insomnia was mediated specifically by reduced overnight resolution of emotional distress. The model outperformed all alternative mediation pathways. The findings suggest that restless REM sleep reflects a process that interferes with the overnight resolution of distress. Its accumulation may promote the development of chronic hyperarousal, giving clinical relevance to the role of REM sleep in emotion regulation in insomnia, depression, and posttraumatic stress disorder.


Vapor-Liquid Sol-Gel Approach to Fabricating Highly Durable and Robust Superhydrophobic Polydimethylsiloxane@Silica Surface on Polyester Textile for Oil-Water Separation.
Large-scale fabrication of superhydrophobic surfaces with excellent durability by simple techniques has been of considerable interest for its urgent practical application in oil-water separation in recent years. Herein, we proposed a facile vapor-liquid sol-gel approach to fabricating highly durable and robust superhydrophobic polydimethylsiloxane@silica surfaces on the cross-structure polyester textiles. Scanning electron microscopy and Fourier transform infrared spectroscopy demonstrated that the silica generated from the hydrolysis-condensation of tetraethyl orthosilicate (TEOS) gradually aggregated at microscale driven by the extreme nonpolar dihydroxyl-terminated polydimethylsiloxane (PDMS(OH)). This led to construction of hierarchical roughness and micronano structures of the superhydrophobic textile surface. The as-fabricated superhydrophobic textile possessed outstanding durability in deionized water, various solvents, strong acid/base solutions, and boiling/ice water. Remarkably, the polyester textile still retained great water repellency and even after ultrasonic treatment for 18 h, 96 laundering cycles, and 600 abrasion cycles, exhibiting excellent mechanical robustness. Importantly, the superhydrophobic polyester textile was further applied for oil-water separation as absorption materials and/or filter pipes, presenting high separation efficiency and great reusability. Our method to construct superhydrophobic textiles is simple but highly efficient; no special equipment, chemicals, or atmosphere is required. Additionally, no fluorinated slianes and organic solvents are involved, which is very beneficial for environment safety and protection. Our findings conceivably stand out as a new tool to fabricate organic-inorganic superhydrophobic surfaces with strong durability and robustness for practical applications in oil spill accidents and industrial sewage emission.


A possible link between the pubertal growth of girls and breast cancer in their daughters.
One hypothesis for the origins of breast cancer is that it is initiated by exposure of developing breast tissue in utero to maternal sex hormones. The sex hormone profile is established at puberty, when it regulates growth of the pelvic bones. The pubertal growth of girls is characterized by broadening and rounding of the pelvis. The maximal width between their iliac crests, the intercristal width, increases more rapidly than in boys. We hypothesized that higher sex hormone concentrations at puberty produce larger intercristal widths, and these are markers of increased breast cancer risk in the next generation. We followed up 6,370 women who were born in Helsinki during 1934-1944, and whose mothers' pelvic bones were measured during routine antenatal care. Women whose mothers had large intercristal widths had higher rates of breast cancer. In those born at or after 40 weeks gestation, the hazard ratio for breast cancer was 3.7 (95% CI: 2.1-6.6) if their mother's intercristal width was greater than 30 cm. Among women born to multiparous mothers this hazard ratio rose to 7.2 (3.4-15.4). Hazard ratios for breast cancer were also higher in the daughters of mothers with round iliac crests. Pelvic bone measurements which increase similarly in girls and boys at puberty did not predict breast cancer. We conclude that the intercristal width, and the roundness of the iliac crests, are markers of mothers' sex hormones, and postulate that high concentrations cause genetic instability in differentiating breast cells in their daughters in utero.


Incidence patterns of primary bone cancer in taiwan (2003-2010): a population-based study.
Primary bone cancer (BC) incidence by age has not been surveyed in Asia. The incidence patterns of nine subtypes of primary BCs registered between 2003 and 2010 were analyzed from Taiwan cancer registry data. More specific analyses were conducted within age groups (Group I: 0-24 years; Group II: 25-59 years; and Group III: 60-85+ years). A total of 1,238 newly diagnosed subjects were registered with an age-standardized incidence rate (ASR) of 6.70 per million person-years. Overall, osteosarcoma (OS: 45 %) was the most common, followed by chondrosarcoma (CS: 18 %), and Ewing sarcoma (ES: 8 %). The percentages of cases and ASRs for age groups I, II, and III were 36.3, 43.0, and 20.7 %, and 7.00, 5.48, and 10.28 per million, respectively. Significant male predilections were observed for all BCs combined, and the CS, chordoma, and malignant ameloblastoma subtypes. Our findings demonstrated an upward trend of 4.8 % per year over the study period, and was more significant for females (6.7 %). A significant increase in trend existed in the incidence of BC among females in Group II, and the incidence of OS and ES among females in Group I. This population-based study has allowed us to confidently define the incidence rates among three age groups of Taiwanese. Despite overall low rates, the upward trend in BC incidence among females may invoke a concern. The results suggest areas for further study into the underlying causes for these cancer trends.


[A Successfully Treated Case of Central Venous Catheter-Related Suppurative Thrombophlebitis of the Right Jugular Vein to the Superior Vena Cava Associated with Esophageal Cancer].
A 63 year-woman was diagnosed with esophageal cancer (Mt. T4N2M0, c-Stage IIIC) after full examination. She underwent chemotherapy with 5-FU+CDDP (FP) through a central venous line (CV). She developed a fever of more than 40°C 15 days after the first course of chemotherapy. We removed the CV owing to suspicion of catheter-related bloodstream infection (CRBSI) and initiated treatment with antibiotics (CFPM). Even so, she experienced swelling from the right cervix to the precordium. We confirmed suppurative thrombophlebitis from the right jugular vein to the superior vena cava by CT and blood culture. After anti-coagulation therapy, venous thrombosis diminished and inflammation was cured. The patient continued to receive chemotherapy, underwent a bypass operation, and completed chemo-radiotherapy successfully while receiving anti-coagulation therapy.


Clinical effects of oleoresin capsicum (pepper spray) on the human cornea and conjunctiva.
To evaluate the effects of oleoresin capsicum (OC) on the human cornea and conjunctiva and to test the effectiveness of topical anesthetics for relief of pain. Prospective, randomized clinical trial. Forty-seven subjects were examined before and at 10 minutes and 1 hour after exposure to pepper spray during a training exercise. Eleven subjects were reexamined at 1 week after exposure. A short, subjective questionnaire was given asking subjects to rate their pain, blurring of vision, and tearing. After exposure, subjects were randomly given a placebo, a topical nonsteroidal antiinflammatory agent, or a topical anesthetic. Visual acuity and corneal sensitivity with a Cochet-Bonnet aesthesiometer (scale of 1-6 cm) was measured and the eyes were examined with a portable slit lamp using fluorescein. Symptoms of pain, blurring of vision, and tearing were recorded in a ranking of 0 to 10. Visual acuity was unaffected by exposure to pepper spray. Corneal sensitivity was reduced from a pretest mean of 5.7 cm to a posttest mean of 0.6 cm 10 minutes after exposure. At 1 hour, the mean corneal sensitivity had recovered to 2.9 cm. Twenty-one percent of eyes had evidence of punctate epithelial erosions, but no corneal abrasions were found. All subjects reported significant pain, blurring of vision, and tearing at 10 minutes that was much improved by 1 hour. Topical flurbiprofen 0.03% improved symptoms in two of 11 subjects, whereas topical proparacaine hydrochloride 0.5% improved symptoms in 16 of 29 eyes. At 1 week after exposure, corneal sensation returned to baseline, and no corneal abnormalities were noted. The predominant symptom after exposure to OC was pain. Topical flurbiprofen was not helpful in reducing symptoms of exposure, whereas topical proparacaine was effective in relieving pain in most subjects. Corneal sensitivity was dramatically reduced at 10 minutes after exposure and was improved after 1 hour. At 1 week, corneal sensation had returned to normal, as had slit-lamp appearance on all subjects examined.


Targeting G Protein-Coupled Receptors by Capture Compound Mass Spectrometry: A Case Study with Sertindole.
Unbiased chemoproteomic profiling of small-molecule interactions with endogenous proteins is important for drug discovery. For meaningful results, all protein classes have to be tractable, including G protein-coupled receptors (GPCRs). These receptors are hardly tractable by affinity pulldown from lysates. We report a capture compound (CC)-based strategy to target and identify GPCRs directly from living cells. We synthesized CCs with sertindole attached to the CC scaffold in different orientations to target the dopamine D2 receptor (DRD2) heterologously expressed in HEK 293 cells. The structure-activity relationship of sertindole for DRD2 binding was reflected in the activities of the sertindole CCs in radioligand displacement, cell-based assays, and capture compound mass spectrometry (CCMS). The activity pattern was rationalized by molecular modelling. The most-active CC showed activities very similar to that of unmodified sertindole. A concentration of DRD2 in living cells well below 100 fmol used as an experimental input was sufficient for unambiguous identification of captured DRD2 by mass spectrometry. Our new CCMS workflow broadens the arsenal of chemoproteomic technologies to close a critical gap for the comprehensive characterization of drug-protein interactions.


Low birth weight and preterm delivery as risk factors for asthma and atopic dermatitis in young adult males.
Gestational factors have been hypothesized to play a role in the susceptibility to asthma and atopic dermatitis. We examined whether fetal growth was associated with asthma and atopic dermatitis separately in a population of 4,795 male conscripts born between 1973 and 1975 in Denmark. The prevalence of asthma was 4.7%. The prevalence odds ratio of asthma in conscripts with a birth weight below 2,501 g was 1.5 (95% confidence interval = 0.7-3.1) compared with conscripts with a birth weight of 3,001-3,500 g, adjusted for gestational age and potential confounders. The adjusted prevalence odds ratio among conscripts born before 34 gestational weeks was 0.8 (95% confidence interval = 0.3-2.0) compared with conscripts born at term. The prevalence of atopic dermatitis was 1.0%. The prevalence odds ratio of atopic dermatitis among those with a birth weight below 2,501 g was 3.0 (95% confidence interval = 0.8-11.9) compared with those whose birth weight was between 3,001 and 3,500 g. Men whose gestational age had been below 34 weeks had an adjusted prevalence odds ratio of 0.3 (95% confidence interval = 0.0-3.1). These findings indicate that fetal growth retardation rather than preterm delivery of male infants is the main gestational factor underlying the associations but does not explain the apparent increase over time of asthma or atopic diseases.


Selective advantages created by codon ambiguity allowed for the evolution of an alternative genetic code in Candida spp.
Several species of the genus Candida decode the standard leucine CUG codon as serine. This and other deviations from the standard genetic code in both nuclear and mitochondrial genomes invalidate the notion that the genetic code is frozen and universal and prompt the questions 'why alternative genetic codes evolved and, more importantly, how can an organism survive a genetic code change?' To address these two questions, we have attempted to reconstruct the early stages of Candida albicans CUG reassignment in the closely related yeast Saccharomyces cerevisiae. These studies suggest that this genetic code change was driven by selection using a molecular mechanism that requires CUG ambiguity. Such codon ambiguity induced a significant decrease in fitness, indicating that CUG reassignment can only be selected if it introduces an evolutionary edge to counteract the negative impact of ambiguity. We have shown that CUG ambiguity induces the expression of a novel set of stress proteins and triggers the general stress response, which, in turn, creates a competitive edge under stress conditions. In addition, CUG ambiguity in S. cerevisiae induces the expression of a number of novel phenotypes that mimic the natural resistance to stress characteristic of C. albicans. The identification of an evolutionary advantage created by CUG ambiguity is the first experimental evidence for a genetic code change driven by selection and suggests a novel role for codon reassignment in the adaptation to new ecological niches.


Neural regulation of cardiovascular response to exercise: Part II.
The changes in circulatory system behavior occurring in response to exercise stress are integrated by the central neural circulatory control system working in cooperation with local regulatory mechanisms. The regulatory events occurring during exercise may be analyzed using a closed-loop regulatory system model of the circulation. In this model, the brain stem acts as the central integration unit determining the set point of the system and the gain of incoming signals from variables such as muscle metabolites and blood gases. Control signals reach the effector organs (heart and peripheral vascular smooth muscle) through sympathetic and parasympathetic motor fibers. Exercise acts on the system as a disturbance, initiating changes in function of effector organs. Local regulatory mechanisms act within the system without passing the central integration unit.


Pathview: an R/Bioconductor package for pathway-based data integration and visualization.
Pathview is a novel tool set for pathway-based data integration and visualization. It maps and renders user data on relevant pathway graphs. Users only need to supply their data and specify the target pathway. Pathview automatically downloads the pathway graph data, parses the data file, maps and integrates user data onto the pathway and renders pathway graphs with the mapped data. Although built as a stand-alone program, Pathview may seamlessly integrate with pathway and functional analysis tools for large-scale and fully automated analysis pipelines. The package is freely available under the GPLv3 license through Bioconductor and R-Forge. It is available at http://bioconductor.org/packages/release/bioc/html/pathview.html and at http://Pathview.r-forge.r-project.org/. firstname.lastname@example.com Supplementary data are available at Bioinformatics online.


Comparison of two sedation techniques in patients undergoing surgical procedures under regional anaesthesia.
Intraoperative comfort and patient satisfaction during surgical procedures under regional anaesthesia can be improved with the use of supplemental intravenous sedation. The authors conducted a study to compare two sedation techniques for surgical procedures performed under regional anaesthesia, i.e., midazolam and pethidine combination compared with midazolam and tramadol combination. Forty adult American Society of Anaesthesiologists (ASA) grade 1-111 patients, aged between 40-65 years undergoing surgery under regional anaesthesia (sub-arachnoid block) were included. The patients were randomly divided into two groups. All patients received standardized premedication, intraoperative monitoring and oxygen therapy. Group A patients received midazolam 0.03 mg/kg followed by pethidine 20 mg intravenously, and group B patients received midazolam 0.03 mg/kg followed by tramadol 20 mg intravenously after the institution of regional anaesthesia. Monitoring included ECG, blood pressure, respiratory rate, oxygen saturation and sedation score. Complications, if any, were recorded. Monitoring was continued during the recovery room stay. All patients were interviewed in the evening and time of ambulation and rating of OR experience was noted. Data analysis showed no significant difference between pethidine and tramadol for all the haemodynamic variables (p = > 0.05). There was also no significant difference in patient's and surgeon's assessment of their experience. Complications and recovery characteristics also did not show any significant difference. Midazolam-tramadol combination may be used as an alternative to midazolam-pethidine combination for sedation during surgical procedures performed under regional anaesthesia.


Chemical proteomics with sulfonyl fluoride probes reveals selective labeling of functional tyrosines in glutathione transferases.
Chemical probes have great potential for identifying functional residues in proteins in crude proteomes. Here we studied labeling sites of chemical probes based on sulfonyl fluorides (SFs) on plant and animal proteomes. Besides serine proteases and many other proteins, SF-based probes label Tyr residues in glutathione transferases (GSTs). The labeled GSTs represent four different GST classes that share less than 30% sequence identity. The targeted Tyr residues are located at similar positions in the promiscuous substrate binding site and are essential for GST function. The high selectivity of SF-based probes for functional Tyr residues in GSTs illustrates how these probes can be used for functional studies of GSTs and other proteins in crude proteomes.


Predicting 24-hour urinary protein excretion in Afro-Caribbean Barbadians by comparing urine protein excretion over different durations versus spot collection.
The gold standard for the determination of proteinuria, an independent risk factor for cardiovascular and renal disease, is the measurement of protein in a 24-hour urine collection. However this method has been shown to be unreliable mainly due to poor compliance of sampling by patients. This study investigates other appropriate means of predicting 24-hour urinary protein excretion in a sample of Afro-Caribbeans in Barbados by assessing the correlation of actual and estimated urinary protein excretion between a 24-hour urine collection sample, 12-hour (AM and PM) and spot (AM and PM) urine collections. A convenient sample of 30 healthy participants of Afro-Caribbean origin between the ages of21 and 55 years was recruited for the study The 24-hour urine samples and anthropometric data were collected as documented in the study s standard clinical procedure. A 24-hour urine sample was collected as two separate 12-hour AM and PM samples. In addition, two spot samples (AM and PM) were taken during each 12-hour sample collection period. Analysis of the urinary protein and creatinine was done with a Roche/Hitachi Modular System (Roche Diagnostics, IN, USA). SPSS version 19 was used to analyse the data to make inferences. Thirty Afro-Caribbean persons participated in the study: 16 females and 14 males. The average age and body mass index (BMI) were 38 +/- 17 years and 25.32+/- 5.98 kg/m2, respectively. The Spearman Rho 's correlation was used to interpret associations of the urinary parameters in 24-hour collected sample and the other samples. The strongest correlation of the protein:creatinine ratio in the 24-hour collected sample to the other samples was observed with the 12-hour AM sample (r = + 0.743, p < O.01)followed by the 12-hour PM sample (r = +0.672, p < 0.01). On analysing gender, the more significant correlations found were among the males for the 12-hour timed samples with r = +0.945, p < 0.01 and r = +0.736, p < 0.01 for the AM and PM samples, respectively. There were very strong correlations between the 24-hour urinary protein excretion and the estimated 24-hour protein excretion from the 12-hour AM and PM samples (r = +0.846, p < 0.01 and r = +0.637, p < 0.01, respectively). Both males and females had the strongest correlation for the estimation of 24-hour protein excretion in the 12-hour AM sample (r = +0.795, p < 0.01 and r = +0.965, p < 0.01, respectively). The use ofa 12-hour timed sample, specifically the morning sample, may be a more convenient way to assess proteinuria in the Afro-Caribbean population. This method allows for a quicker assessment of proteinuria which not only allows earlier diagnosis of renal disease but may also reduce the clinical cost of the disease s management.


Roles of Pax-6 in murine diencephalic development.
Pax-6 is one of the earliest regulatory genes to be expressed in the diencephalon. We tested whether normal Pax-6 protein is required for early diencephalic development by examining morphology, precursor proliferation and patterns of regulatory gene expression in the embryonic diencephalon of Small-eye mice (Pax-6 mutants). In Small-eye mice, diencephalic morphology was abnormal at all the embryonic ages studied (days 10.5, 12.5 and 14.5). Regional differences in diencephalic cell density were lost, the diencephalon/mesencephalon boundary was unclear and the third ventricle was enlarged. We estimated diencephalic proliferative rates after labelling with bromodeoxyuridine and found that they were abnormally low in mutants aged embryonic day 10.5. In older mutants, the diencephalon contained fewer cells than normal. In wild-type E14.5 diencephalon, Pax-6, Dlx-2 and Wnt-3 are expressed in discrete regions along the rostrocaudal and dorsoventral axes. In situ hybridizations for these genes in E14.5 Small-eye mice revealed discrete zones of diencephalic expression that had similar relative positions to those in wild-type mice. Some differences of detail in their expression were seen: Pax-6 had an expanded rostral domain of expression and an abnormally indistinct caudal boundary; Dlx-2 had a diffuse, rather than a sharp, caudal boundary of expression; the normally high dorsal midline expression of Wnt-3 was lost. We conclude that normal expression of Pax-6 is required for the correct regulation of diencephalic precursor proliferation. Pax-6 may also control some aspects of diencephalic differentiation, but its mutation in Small-eye mice does not preclude the development of a degree of diencephalic regionalization resembling that in normal mice.


Deconvolution and quantification of hydrocarbon-like and oxygenated organic aerosols based on aerosol mass spectrometry.
A new technique has been developed to deconvolve and quantify the mass concentrations of hydrocarbon-like and oxygenated organic aerosols (HOA and OOA) using highly time-resolved organic mass spectra obtained with an Aerodyne Aerosol Mass Spectrometer (AMS). This technique involves a series of multivariate linear regressions that use mass-to-charge ratios (ml/s) 57 (mostly C4H9+) and 44 (mostly CO2+)-the identified AMS mass spectral tracers for HOA and OOA, respectively-as the initial principal components. Two algorithms have been developed: algorithm 1 is based solely on m/z 44 and m/z 57, and algorithm 2 is an iterative procedure expanded from algorithm 1. This technique was applied to the AMS organic aerosol data acquired at the EPA Pittsburgh Supersite during September 2002. The reconstructed organic concentrations (= HOA + OOA) agree well with the measured values (r2 = 0.997, slope = 0.998), and the reconstructed organic data matrix (size = 3199 time steps x 300 m/z's) explains 99% of the variance in the measured time series. In addition, the extracted mass spectrum of HOA shows high similarity to those of diesel exhaust, lubricating oil, and freshly emitted traffic aerosols observed in urban areas, while the spectrum of OOA closely resembles those of aged organic aerosols sampled in rural areas and also shows similarity with the spectrum of fulvic acid- a humic-like substance that is ubiquitous in the environment and has previously been used as an analogue to represent polyacid components found in highly processed and oxidized atmospheric organic aerosols. There is evidence for the presence of a third component, although its contribution to the total organic signal appears to be small in this study. The most important result is that m/z 44 and m/z 57 are reliable AMS mass spectral "markers" that provide the "first guess" for algorithm 2 which allows the quantitative description of the organic aerosol concentration and mass spectra over a period of 16 days in a major urban area and allows the extraction of mass spectra of OOA and HOA that can be interpreted chemically. These findings indicate the potential of performing organic source apportionment on the basis of total particle mass, rather than on the basis of organic tracer compounds that contribute a small fraction of this mass.


Effects of menopause and high-intensity training on insulin sensitivity and muscle metabolism.
To investigate peripheral insulin sensitivity and skeletal muscle glucose metabolism in premenopausal and postmenopausal women, and evaluate whether exercise training benefits are maintained after menopause. Sedentary, healthy, normal-weight, late premenopausal (n = 21), and early postmenopausal (n = 20) women were included in a 3-month high-intensity exercise training intervention. Body composition was assessed by magnetic resonance imaging and dual-energy x-ray absorptiometry, whole body glucose disposal rate (GDR) by hyperinsulinemic euglycemic clamp (40 mU/m/min), and femoral muscle glucose uptake by positron emission tomography/computed tomography, using the glucose analog fluorodeoxyglucose, expressed as estimated metabolic rate (eMR). Insulin signaling was investigated in muscle biopsies. Age difference between groups was 4.5 years, and no difference was observed in body composition. Training increased lean body mass (estimate [95% confidence interval] 0.5 [0.2-0.9] kg, P < 0.01) and thigh muscle mass (0.2 [-0.1 to 0.6] kg, P < 0.01), and decreased fat percentage (1.0 [0.5-1.5]%, P < 0.01) similarly in the two groups. The postmenopausal women had lower eMR in vastus lateralis muscle than the premenopausal women (-14.0 [-26.0 to -2.0] μmol/min/kg, P = 0.02), and tended to have lower eMR in femoral muscles (-11.2 [-22.7 to 0.4] μmol/min/kg, P = 0.06), and also GDR (-59.3 [-124.8 to 6.3] mg/min, P = 0.08), but increased similarly in both groups with training (eMR vastus lateralis muscle: 27.8 [19.6-36.0] μmol/min/kg, P < 0.01; eMR femoral muscle: 20.0 [13.1-26.7] μmol/min/kg, P < 0.01, respectively; GDR: 43.6 [10.4-76.9] mg/min, P = 0.01). Potential mechanisms underlying the training-induced increases in insulin sensitivity included increased expression of hexokinase (19.2 [5.0-24.7] AU, P = 0.02) and glycogen synthase (32.4 [15.0-49.8] AU, P < 0.01), and also increased insulin activation of Akt2 (20.6 [3.4-29.0], P = 0.03) and dephosphorylation of glycogen synthase (-41.8 [-82.9 to -0.7], P = 0.05). Insulin sensitivity was reduced in early postmenopausal women. However, postmenopausal women increased peripheral insulin sensitivity, skeletal muscle insulin-stimulated glucose uptake, and skeletal muscle mass to the same extent as premenopausal women after 3 months of high-intensity exercise training.


Development of classification and regression based QSAR models to predict rodent carcinogenic potency using oral slope factor.
Carcinogenicity is among the toxicological endpoints posing the highest concern for human health. Oral slope factors (OSFs) are used to estimate quantitatively the carcinogenic potency or the risk associated with exposure to the chemical by oral route. Regulatory agencies in food and drug administration and environmental protection are employing quantitative structure-activity relationship (QSAR) models to fill the data gaps related with properties of chemicals affecting the environment and human health. In this background, we have developed quantitative structure-carcinogenicity regression models for rodents based on the carcinogenic potential of 70 chemicals with wide diversity of molecular structures, spanning a large number of chemical classes and biological mechanisms. All the developed models have been assessed according to the Organization for Economic Cooperation and Development (OECD) principles for the validation of QSAR models. We have also attempted to develop a carcinogenicity classification model based on Linear Discriminant Analysis (LDA). Developed regression and LDA models are rigorously validated internally as well as externally. Our in silico studies make it possible to obtain a quantitative interpretation of the structural information of carcinogenicity along with identification of the discriminant functions between lower and higher carcinogenic compounds by LDA. Pharmacological distribution diagrams (PDDs) are used as a visualizing technique for the identification and selection of chemicals with lower carcinogenicity. Constructive, informative and comparable interpretations have been observed in both cases of classification and regression based modeling.


Total synthesis of biselide A.
A total synthesis of the marine macrolide biselide A is described that relies on an enantiomerically enriched α-chloroaldehyde as the sole chiral building block. Several strategies to construct the macrocycle are presented including a macrocyclic Reformatsky reaction that ultimately provides access to the natural product in a longest linear sequence of 18 steps. Biological testing of synthetic biselide A suggests this macrolide disrupts cell division through a mechanism related to the regulation of microtubule cytoskeleton organization. Overall, this concise synthesis and insight gained into the mechanism of action should inspire medicinal chemistry efforts directed at structurally related anticancer marine macrolides.


Allium Extract Implements Weaned Piglet's Productive Parameters by Modulating Distal Gut Microbiota.
Antimicrobial resistance (AMR) has risen as a global threat for human health. One of the leading factors for this emergence has been the massive use of antibiotics growth-promoter (AGPs) in livestock, enhancing the spread of AMR among human pathogenic bacteria. Thus, several alternatives such as probiotics, prebiotics, or phytobiotics have been proposed for using in animal feeding to maintain or improve productive levels while diminishing the negative effects of AGPs. Reducing the use of antibiotics is a key aspect in the pig rearing for production reasons, as well as for the production of high-quality pork, acceptable to consumers. Here we analyze the potential use of Allium extract as an alternative. In this study, weaned piglets were fed with Allium extract supplementation and compared with control and antibiotic (colistin and zinc oxide) treated piglets. The effects of Allium extract were tested by analyzing the gut microbiome and measuring different productive parameters. Alpha diversity indices decreased significantly in Allium extract group in caecum and colon. Regarding beta diversity, significant differences between treatments appeared only in caecum and colon. Allium extract and antibiotic piglets showed better values of body weight (BW), average daily weight gain (ADG), and feed conversion ratio (FCR) than control group. These results indicate that productive parameters can be implemented by modifying the gut microbiota through phytobiotics such as Allium extract, which will drive to drop the use of antibiotics in piglet diet.


Interior design: how plant pathogens optimize their living conditions.
Pathogens use effectors to suppress host defence mechanisms, promote the derivation of nutrients, and facilitate infection within the host plant. Much is now known about effectors that target biotic pathways, particularly those that interfere with plant innate immunity. By contrast, an understanding of how effectors manipulate nonimmunity pathways is only beginning to emerge. Here, we focus on exciting new insights into effectors that target abiotic stress adaptation pathways, tampering with key functions within the plant to promote colonization. We critically assess the role of various signalling agents in linking different pathways upon perturbation by pathogen effectors. Additionally, this review provides a summary of currently known bacterial, fungal, and oomycete pathogen effectors that induce biotic and abiotic stress responses in the plant, as a first step towards establishing a comprehensive picture for linking effector targets to pathogenic lifestyles.


GF-15, a novel inhibitor of centrosomal clustering, suppresses tumor cell growth in vitro and in vivo.
In contrast to normal cells, malignant cells are frequently aneuploid and contain multiple centrosomes. To allow for bipolar mitotic division, supernumerary centrosomes are clustered into two functional spindle poles in many cancer cells. Recently, we have shown that griseofulvin forces tumor cells with supernumerary centrosomes to undergo multipolar mitoses resulting in apoptotic cell death. Here, we describe the characterization of the novel small molecule GF-15, a derivative of griseofulvin, as a potent inhibitor of centrosomal clustering in malignant cells. At concentrations where GF-15 had no significant impact on tubulin polymerization, spindle tension was markedly reduced in mitotic cells upon exposure to GF-15. Moreover, isogenic cells with conditional centrosome amplification were more sensitive to GF-15 than parental controls. In a wide array of tumor cell lines, mean inhibitory concentrations (IC(50)) for proliferation and survival were in the range of 1 to 5 μmol/L and were associated with apoptotic cell death. Importantly, treatment of mouse xenograft models of human colon cancer and multiple myeloma resulted in tumor growth inhibition and significantly prolonged survival. These results show the in vitro and in vivo antitumor efficacy of a prototype small molecule inhibitor of centrosomal clustering and strongly support the further evaluation of this new class of molecules.


Making sense of evidence.
There is a constant stream of information emanating from psychiatric research. The challenge for the clinician to keep abreast of the latest research findings is huge. However, even more daunting is the task of making sense of the sometimes conflicting data. In this paper, we provide some examples of evidence that seemed to have been accepted with relatively little critical examination. We discuss how evidence may look plausible on the surface either because contextual factors have not been considered or because they fit into a pre-determined world view. We argue that, in the end, the process of making sense of evidence is not straightforward and that professional biases and social prejudices often influence decisions.


Sex 'n' drugs 'n' rock 'n' roll: the meaning and social consequences of pubertal timing.
This is a brief review of the normal changes in adolescent behaviour and the interplay between biology and social factors that occur at and around puberty, in an attempt to explain when this transition may become problematic The onset of puberty is a biological marker for an individual's transition from a non-reproductive to a reproductive state. Adolescence is a normal developmental transition associated with clearly visible physical changes, reorganization and pruning of neuronal circuits in the brain and the occurrence of new behaviours and interests. It is a time when new life tasks (orientation towards peers of the other sex, romantic and sexual involvement and mastering an educational career) need to be mastered. Parent-child conflict increases and becomes more intense as the adolescent struggles for more independence while still requiring support. These normal changes can become problematic if biological and social expectations diverge e.g. entering puberty very early or very late. While early pubertal onset in boys is likely to have beneficial effects, in girls precocious pubertal timing may have a negative impact on body-image, affect (or emotional well-being) and sex-role expectations. Other individual biological predispositions and genetic endowment may interact with social factors (e.g. peers, parenting style, neighbourhood) making adolescence either an adaptive or a challenging transition. There is a lack of sufficiently large longitudinal studies that have been able to study this interaction between genetics, biology and social environment on adolescent development. The Avon Longitudinal Study of Parents and Children (ALSPAC) cohort provides a unique opportunity to investigate the impact of pubertal timing on social behaviour. Planned assessments and concepts are outlined.


Examining the architecture of cellular computing through a comparative study with a computer.
The computer and the cell both use information embedded in simple coding, the binary software code and the quadruple genomic code, respectively, to support system operations. A comparative examination of their system architecture as well as their information storage and utilization schemes is performed. On top of the code, both systems display a modular, multi-layered architecture, which, in the case of a computer, arises from human engineering efforts through a combination of hardware implementation and software abstraction. Using the computer as a reference system, a simplistic mapping of the architectural components between the two is easily detected. This comparison also reveals that a cell abolishes the software-hardware barrier through genomic encoding for the constituents of the biochemical network, a cell's "hardware" equivalent to the computer central processing unit (CPU). The information loading (gene expression) process acts as a major determinant of the encoded constituent's abundance, which, in turn, often determines the "bandwidth" of a biochemical pathway. Cellular processes are implemented in biochemical pathways in parallel manners. In a computer, on the other hand, the software provides only instructions and data for the CPU. A process represents just sequentially ordered actions by the CPU and only virtual parallelism can be implemented through CPU time-sharing. Whereas process management in a computer may simply mean job scheduling, coordinating pathway bandwidth through the gene expression machinery represents a major process management scheme in a cell. In summary, a cell can be viewed as a super-parallel computer, which computes through controlled hardware composition. While we have, at best, a very fragmented understanding of cellular operation, we have a thorough understanding of the computer throughout the engineering process. The potential utilization of this knowledge to the benefit of systems biology is discussed.


The availability and usage frequency of real time ultrasound by physiotherapists in South Australia: an observational study.
Real time ultrasound (RTUS) is an emerging imaging modality in physiotherapy. Anecdotal evidence suggests that it is being used as an assessment and biofeedback tool for various deep core stabilizing muscles. However, how and why physiotherapists use RTUS in the clinical setting has not yet been reported. Therefore, this study aimed to establish the availability and usage frequency of RTUS by physiotherapists in South Australia. In addition, the study aimed to describe how physiotherapists were using RTUS and how they were educated in its use. A questionnaire was developed and mailed to all physiotherapists registered with the Physiotherapy Board of South Australia (n = 1328) between February and March 2007. A response rate of 50% was achieved with 664 completed usable questionnaires returned. At the current time, only a small proportion of respondents used RTUS (11.6%), while slightly more had access to a machine (19.4%). RTUS was used most commonly for assessment (88.3%) and biofeedback (87.0%) of the abdominal (94.7%), pelvic floor (72.7%) and multifidus (54.5%) muscles. Of all respondents, 26.7% had trained in its use with most completing two hours or less of training. This is the first published study to describe how and why physiotherapists are using RTUS in clinical practice. RTUS appears to be a relatively uncommon modality potentially limited by insufficient access to equipment and educational opportunities. The findings highlight a greater need for education and training in the use of RTUS for physiotherapy practice.


Organic and metallic pollutants in water treatment and natural wetlands: a review.
A literature review shows that more than 500 compounds occur in wetlands, and also that wetlands are suitable for removing these compounds. There are, however, obvious pitfalls for treatment wetlands, the most important being the maintenance of the hydraulic capacity and the detention time. Treatment wetlands should have an adapted design to target specific compounds. Aquatic plants and soils are suitable for wastewater treatment with a high capacity of removing nutrients and other substances through uptake, sorption and microbiological degradation. The heavy metals Cd, Cu, Fe, Ni and Pb were found to exceed limit values. The studies revealed high values of phenol and SO(4). No samples showed concentrations in sediments exceeding limit values, but fish samples showed concentrations of Hg exceeding the limit for fish sold in the European Union (EU). The main route of metal uptake in aquatic plants was through the roots in emergent and surface floating plants, whereas in submerged plants roots and leaves take part in removing heavy metals and nutrients. Submerged rooted plants have metal uptake potential from water as well as sediments, whereas rootless plants extracted metals rapidly only from water. Caution is needed about the use of SSF CWs (subsurface flow constructed wetlands) for the treatment of metal-contaminated industrial wastewater as metals are shifted to another environmental compartment, and stable redox conditions are required to ensure long-term efficiency. Mercury is one of the most toxic heavy metals and wetlands have been shown to be a source of methylmercury. Methyl Hg concentrations are typically approximately 15% of Hgt (total mercury). In wetlands polycyclic aromatic hydrocarbons (PAH), bisphenol A, BTEX, hydrocarbons including diesel range organics, glycol, dichlorodiphenyltrichloroethane (DDT), polychlorinated biphenyls (PCB), cyanide, benzene, chlorophenols and formaldehyde were found to exceed limit values. In sediments only PAH and PCB were found exceeding limit values. The pesticides found above limit values were atrazine, simazine, terbutylazine, metolachlor, mecoprop, endosulfan, chlorfenvinphos and diuron. There are few water quality limit values of these compounds, except for some well-known endocrine disrupters such as nonylphenol, phtalates, etc.


Anterior approach to thoracic and lumbar spine lesions: results in 145 consecutive cases.
The authors report on a series of 145 consecutive patients with different types of spine lesions surgically treated via an anterior approach (AA) at the thoracic and lumbar levels during the past 10 years. Indications, techniques, and surgical results are described. This series included 92 patients with fractures, 30 with neoplasms, 13 with thoracic disc hernias, and 10 with spinal infections. Based on the lesion to be addressed, the AA was used for lesion excision, corpectomy, vertebral body reconstruction with cages, realignment, and/or plating or screwing. The approach was extracavitary in 55 patients and intracavitary in 90. In 126 patients (86.8%), neural decompression and spine stabilization were achieved via a stand-alone AA (SA-AA), whereas 19 patients (13.1%) were treated using a 2-stage anteroposterior approach. This circumferential approach was reserved for select cases of severe traumatic dislocation, particular types of tumors, or specific anatomical locations. The authors developed a simple neuronavigation-based method of identifying the severely injured patients who were eligible for the SA-AA by evaluating the angle of lateral dislocation. There were no deaths and no instances of major surgery-related morbidity. Minor morbidity was almost always transitory and was reported in 13 patients (8.9%). Neurological improvement was reported in 20% of injured patients with a preoperative incomplete lesion. Postoperatively, all patients were able to stand or at least sit without load pain. During the follow-up (mean +/- standard deviation 3.8 +/- 2.4 years), there were no cases of failure, fracture, dislocation, or bending of the anterior instrumentation, and the rate of pseudarthrosis was 0%. The anterior route provides direct access to most spine diseases and allows optimal neural decompression and the possibility of adequate realignment and strong reconstruction/fixation. Stability of the vertebral column is achieved, resolution of clinical pain is rapid and almost complete, and the rate of surgical complications is very low. The authors assert that the SA-AA offers so many advantages and has such good results that the 2-stage anteroposterior approach can be reserved for a minority of select cases and that the time for using the posterior approach alone is over.


Interaction between the ADH1B*3 allele and drinking motives on alcohol use among Black college students.
Black young adults have lower rates of alcohol use than other racial groups. Genetic factors may protect against drinking. Specifically, the ADH1B*3 allele is present almost exclusively in Black populations and has been protective against alcohol use and alcohol use disorder. The protective effects of the ADH1B*3 allele, however, may differ as a function of alcohol-promoting cognitions. The current study examined whether ADH1B*3 moderated relations of drinking motives with alcohol consumption among Black college drinkers. Participants were 241 undergraduate students of self-identified Black race (mean age = 20 years; 66% female) who reported consuming alcohol at least once in the past 30 days. ADH1B*3 was not significantly associated with drinking motives or drinking behaviors. However, significant, albeit small, interaction effects of ADH1B*3 with drinking motives on drinking behavior were found; the presence of an ADH1B*3 allele protected against greater drinking quantity among students with high social motives (incidence rate ratio [IRR] = 0.95, 95% CI [0.92, 0.99]) and against frequent drinking among students with low coping motives (IRR = 1.06, 95% CI [1.01, 1.11]). These findings represent a novel demonstration of genetic modulation of alcohol-related cognitions within Black college drinkers, although replication is needed. Results represent an initial step toward better characterizing individual differences in associations of drinking motives with drinking behavior, with potential implications for interventions aimed at motivational processes in alcohol use.


Broadband enhanced backscattering spectroscopy of strongly scattering media.
We report on a new experimental method for enhanced backscattering spectroscopy (EBS) of strongly scattering media over a bandwidth from 530-1000 nm. The instrument consists of a supercontinuum light source and an angle-dependent detection system using a fiber-coupled grating spectrometer. Using a combination of two setups, the backscattered intensity is obtained over a large angular range and using circularly polarized light. We present broadband EBS of a TiO(2) powder and of a strongly scattering porous GaP layer. In combination with theoretical model fits, the EBS system yields the optical transport mean free path over the available spectral window.


Dissecting the ER-associated degradation of a misfolded polytopic membrane protein.
It remains unclear how misfolded membrane proteins are selected and destroyed during endoplasmic reticulum-associated degradation (ERAD). For example, chaperones are thought to solubilize aggregation-prone motifs, and some data suggest that these proteins are degraded at the ER. To better define how membrane proteins are destroyed, the ERAD of Ste6p(*), a 12 transmembrane protein, was reconstituted. We found that specific Hsp70/40s act before ubiquitination and facilitate Ste6p(*) association with an E3 ubiquitin ligase, suggesting an active role for chaperones. Furthermore, polyubiquitination was a prerequisite for retrotranslocation, which required the Cdc48 complex and ATP. Surprisingly, the substrate was soluble, and extraction was independent of a ubiquitin chain extension enzyme (Ufd2p). However, Ufd2p increased the degree of ubiquitination and facilitated degradation. These data indicate that polytopic membrane proteins can be extracted from the ER, and define the point of action of chaperones and the requirement for Ufd2p during membrane protein quality control.


Systemic sclerosis sine scleroderma: a multicenter study of 1417 subjects.
To describe the clinical and serological features of systemic sclerosis sine scleroderma (ssSSc) in a multicentered SSc cohort. Data from 1417 subjects in the Canadian Scleroderma Research Group registry were extracted to identify subjects with ssSSc, defined as SSc diagnosed by an expert rheumatologist, but without any sclerodactyly or skin involvement prior to baseline study visit or during followup. Clinical and serological features of ssSSc subjects were compared to limited (lcSSc) and diffuse cutaneous SSc (dcSSc) subjects. At the first registry visit, only 57 subjects (4.0%) were identified as having ssSSc. Of these, 30 (2.1%) were reclassified as lcSSc within 1.9 years. Thus, only 27 ssSSc subjects (1.9%) remained, with mean followup of 2.4 years. Clinical profiles of ssSSc were generally similar or milder compared to lcSSc, and milder than dcSSc, including rates of interstitial lung disease (25.9% ssSSc, 25.4% lcSSc, 40.3% dcSSc). Patients with ssSSc had serological profiles similar to those with lcSSc, including high rates of anticentromere antibodies (50.0% ssSSc, 47.5% lcSSc, 12.1% dcSSc), and low rates of antitopoisomerase I (16.7% ssSSc, 7.0% lcSSc, 21.8% dcSSc) and anti-RNA polymerase III (0 ssSSc, 11.1% lcSSc, 34.9% dcSSc). The condition ssSSc is rare and resembles lcSSc. These observations suggest that ssSSc is most likely a forme fruste of lcSSc, and that the absence of skin involvement may in part be related to misclassification arising from early or subtle skin involvement. There is little evidence to consider ssSSc as a distinct clinical or serological subset of SSc.


Halo-free Phase Contrast Microscopy.
We present a new approach for retrieving halo-free phase contrast microscopy (hfPC) images by upgrading the conventional PC microscope with an external interferometric module, which generates sufficient data for reversing the halo artifact. Acquiring four independent intensity images, our approach first measures haloed phase maps of the sample. We solve for the halo-free sample transmission function by using a physical model of the image formation under partial spatial coherence. Using this halo-free sample transmission, we can numerically generate artifact-free PC images. Furthermore, this transmission can be further used to obtain quantitative information about the sample, e.g., the thickness with known refractive indices, dry mass of live cells during their cycles. We tested our hfPC method on various control samples, e.g., beads, pillars and validated its potential for biological investigation by imaging live HeLa cells, red blood cells, and neurons.


The natural history of Teflon catheter-associated phlebitis in children.
During a prospective evaluation of intravenous therapy with peripheral Teflon catheters in children, we found 30 episodes of phlebitis (10.4%). This rate is less than that reported in adults. Catheter colonization was not related to phlebitic episodes, and catheter-related infections did not occur. No patient's hospital course was prolonged because of phlebitis. Thirty percent of the episodes developed after the catheter was removed, and premonitory symptoms were not helpful in predicting the onset of phlebitis. Factors associated with an increased phlebitic risk were parenteral nutrition, administration of nafcillin sodium or aminoglycosides, and patient age. Parenteral nutrition prolonged the course of phlebitis. No factors hastened the onset of phlebitis. The duration of cannulation was not significantly related to phlebitis, suggesting that in some children the catheters can remain in place longer than 72 hours.


Loss of allergen 1 confers a hypervirulent phenotype that resembles mucoid switch variants of Cryptococcus neoformans.
Microbial survival in a host is usually dependent on the ability of a pathogen to undergo changes that promote escape from host defense mechanisms. The human-pathogenic fungus Cryptococcus neoformans undergoes phenotypic switching in vivo that promotes persistence in tissue. By microarray and real-time PCR analyses, the allergen 1 gene (ALL1) was found to be downregulated in the hypervirulent mucoid switch variant, both during logarithmic growth and during intracellular growth in macrophages. The ALL1 gene encodes a small cytoplasmic protein that is involved in capsule formation. Growth of an all1Delta gene deletion mutant was normal. Similar to cells of the mucoid switch variant, all1Delta cells produced a larger polysaccharide capsule than cells of the smooth parent and the complemented strain produced, and the enlarged capsule inhibited macrophage phagocytosis. The mutant exhibited a modest defect in capsule induction compared to all of the other variants. In animal models the phenotype of the all1Delta mutant mimicked the hypervirulent phenotype of the mucoid switch variant, which is characterized by decreased host survival and elevated intracranial pressure. Decreased survival is likely the result of both an ineffective cell-mediated immune response and impaired phagocytosis by macrophages. Consequently, we concluded that, unlike loss of most virulence-associated genes, where loss of gene function results in attenuated virulence, loss of the ALL1 gene enhances virulence by altering the host-pathogen interaction and thereby impairing clearance. Our data identified the first cryptococcal gene associated with elevated intracranial pressure and support the hypothesis that an environmental opportunistic pathogen has modified its virulence in vivo by epigenetic downregulation of gene function.


Confidential Care for Adolescents in the U.S. Health Care System.
In providing care for adolescents, maintaining confidentiality should be considered a human right and an evidence-based component of quality care. Unfortunately, complexities in the U.S. legal and health care systems have created a setting in which confidential care is inaccessible to many adolescent patients. Federal laws provide a minimum standard for confidentiality protections, but variations in state legislation relating to minor consent, special health care services, and confidentiality exemptions create large variability in adolescent confidentiality rights across the country. In certain contexts, such as consensual sexual activity, legal provisions may not align with professional ethical standards in adolescent care. In addition, contemporary clinical and administrative issues related to the electronic health record and health care financing also threaten to breach confidentiality. Although further research is necessary, providers and institutions are already well positioned to broaden protections by implementing best practices in training and education, workflow, and medical records related to adolescent confidentiality.


Nab-Paclitaxel Followed by 5-Fluorouracil, Epirubicin and Cyclophosphamide in Neoadjuvant Chemotherapy for Resectable Breast Cancer: A Phase II Trial.
The aim of this phase II study was to evaluate combined nab-paclitaxel (nab-PTX) with sequential anthracycline-based therapy as a neoadjuvant chemotherapy. We enrolled 41 patients with advanced breast cancer (stage IIA - IIIC). All patients were to receive three-weekly nab-PTX (260 mg/m2) for four cycles followed by three-weekly 5-fluorouracil, epirubicin and cyclophosphamide (FEC) for four cycles. Trastuzumab administration was permitted in human epidermal growth factor receptor 2 (HER2)-positive patients. The overall pathological complete response (pCR) rate was 24% (10 of 41). In patients with luminal A, luminal B (HER2-), luminal B (HER2+), triple-negative and HER2, the pCR rates were 0% (0/2), 7% (1/14), 42% (3/7), 25% (4/16) and 100% (2/2), respectively. The most significant toxicities of nab-PTX were grade 2/3 peripheral sensory neuropathy (24%) and grade 3/4 neutropenia (26%). Febrile neutropenia was not observed in any patient. The most significant toxicities of FEC were grade 3/4 neutropenia (24%) and grade 3 febrile neutropenia (9%). One patient died of sepsis secondary to pneumonia during FEC treatment. Neoadjuvant chemotherapy using nab-PTX with trastuzumab every 3 weeks followed by FEC was suitably tolerated and associated with a high pCR rate of 55% for patients with HER2-positive breast cancer.


Occurrence and metabolic activity of organisms under the ross ice shelf, antarctica, at station j9.
Seawater samples below the Ross Ice Shelf were collected through an access hole at J9, approximately 400 kilometers from the Ross Sea, Antarctica. The 237-meter water column had sparse populations of bacteria (8.7 x 10(6) to 1.2 x 10(7) per liter), microplankters (10(2) to 10(3) per cubic meter), and zooplankters (10 to 20 per cubic meter) at the depths studied. Microbial biomass estimates from cellular adenosine 5'-triphosphate measurements were very low (10 to 150 nanograms of carbon per liter), comparable with values for the abyssal ocean. Microbial populations assimilated tritiated D-glucose, thymidine, uridine, and adenosine triphosphate at extremely low rates, comparable with deep-sea heterotrophic populations. Sediment samples had 10(7) to 10(8) bacteria per gram (dry weight), which were metabolically active as shown by respiration of uniformly labeled D-[(14)C]glucose. From this study it cannot be determined whether these organisms in the water column and sediments constitute a functioning food web.


Grooming reciprocity in female tibetan macaques macaca thibetana.
Grooming among nonhuman primates is widespread and may represent an important service commodity that is exchanged within a biological marketplace. In this study, using focal animal sampling methods, we recorded grooming relationships among 12 adult females in a free-ranging group of Tibetan macaques (Macaca thibetana) at Huangshan, China, to determine the influence of rank and kinship on grooming relationships, and whether females act as reciprocal traders (exchange grooming received for grooming given) or interchange traders (interchange grooming for social tolerance or other commodities). The results showed that: (1) grooming given was positively correlated with grooming received; (2) kinship did not exert a significant influence on grooming reciprocity; and (3) grooming reciprocity occurred principally between individuals of adjacent rank; however, when females of different rank groomed, females tended to groom up the hierarchy (lower ranking individuals groomed higher ranking individuals more than vice versa). Our results support the contention that both grooming reciprocity and the interchange of grooming for tolerance represent important social tactics used by female Tibetan macaques.


Contextual regulation of pancreatic cancer stem cell phenotype and radioresistance by pancreatic stellate cells.
Progression of pancreatic ductal adenocarcinoma (PDAC) is promoted by desmoplasia induced by pancreatic stellate cells (PSC). Contributory to this progression is epithelial mesenchymal transition (EMT), which shares many characteristics with the cancer stem cell (CSC) hypothesis. We investigated the role of these processes on the radioresponse and tumorigenicity of pancreatic cancer cells. We used an in vitro sphere model and in vivo xenograft model to examine the role of PSC in EMT and CSC processes. We demonstrated that PSC enhanced the CSC phenotype and radioresistance of pancreatic cancer cells. Furthermore, the expression of several EMT and CSC markers supported enhanced processes in our models and that translated into remarkable in vivo tumorigenicity. Multi-dose TGFβ neutralizing antibody inhibited the EMT and CSC processes, sensitized cells to radiation and reduced in vivo tumorigenicity. A proteomic screen identified multiple novel factors that were regulated by PSC in pancreatic cells. These results are critical in highlighting the role of PSC in tumor progression and radioresistance by manipulating the EMT and CSC processes. TGFβ and the novel factors identified are important targets for better therapeutic outcome in response to PSC mediated mechanisms.


Diabetic retinopathy: current understanding, mechanisms, and treatment strategies.
Diabetic retinopathy (DR) causes significant visual loss on a global scale. Treatments for the vision-threatening complications of diabetic macular edema (DME) and proliferative diabetic retinopathy (PDR) have greatly improved over the past decade. However, additional therapeutic options are needed that take into account pathology associated with vascular, glial, and neuronal components of the diabetic retina. Recent work indicates that diabetes markedly impacts the retinal neurovascular unit and its interdependent vascular, neuronal, glial, and immune cells. This knowledge is leading to identification of new targets and therapeutic strategies for preventing or reversing retinal neuronal dysfunction, vascular leakage, ischemia, and pathologic angiogenesis. These advances, together with approaches embracing the potential of preventative or regenerative medicine, could provide the means to better manage DR, including treatment at earlier stages and more precise tailoring of treatments based on individual patient variations.


Opportunities and challenges in the subsoil: pathways to deeper rooted crops.
Greater exploitation of subsoil resources by annual crops would afford multiple benefits, including greater water and N acquisition in most agroecosystems, and greater sequestration of atmospheric C. Constraints to root growth in the subsoil include soil acidity (an edaphic stress complex consisting of toxic levels of Al, inadequate levels of P and Ca, and often toxic levels of Mn), soil compaction, hypoxia, and suboptimal temperature. Multiple root phenes under genetic control are associated with adaptation to these constraints, opening up the possibility of breeding annual crops with root traits improving subsoil exploration. Adaptation to Al toxicity, hypoxia, and P deficiency are intensively researched, adaptation to soil hardness and suboptimal temperature less so, and adaptations to Ca deficiency and Mn toxicity are poorly understood. The utility of specific phene states may vary among soil taxa and management scenarios, interactions which in general are poorly understood. These traits and issues merit research because of their potential value in developing more productive, sustainable, benign, and resilient agricultural systems.


Staff experiences of enhanced recovery after surgery: systematic review of qualitative studies.
To conduct a systematic review of qualitative studies which explore health professionals' experiences of and perspectives on the enhanced recovery after surgery (ERAS) pathway. Systematic review of qualitative literature using a qualitative content analysis. Literature includes the experiences and views of a wide range of multidisciplinary team and allied health professional staff, to incorporate a diverse range of clinical and professional perspectives. PsycINFO, Medline, CINAHL and PubMed were searched in May 2017. The searches included relevant qualitative studies across a range of healthcare contexts. We included studies published from 2000 to 2017, as an appropriate time frame to capture evidence about ERAS after implementation in the late 1990s. Only studies published in the English language were included, and we included studies that explicitly stated that they used qualitative approaches. Literature searches were conducted by the first author and checked by the second author: both contributed to the extraction and analysis of data. Studies identified as relevant were assessed for eligibility using the Critical Appraisal Skills Programme guidance. Eight studies were included in the review, including studies in six countries and in four surgical specialties. Included studies focus on health professionals' experiences of ERAS before, during and after implementation in colorectal surgery, gastrointestinal surgery, abdominal hysterectomy and orthopaedics. Five main themes emerged in the analysis: communication and collaboration, resistance to change, role and significance of protocol-based care, and knowledge and expectations. Professionals described the importance of effective multidisciplinary team collaboration and communication, providing thorough education to staff and patients, and appointing a dedicated champion as means to implement and integrate ERAS pathways successfully. Evidence-based guidelines were thought to be useful for improvements to patient care by standardising practices and reducing treatment variations, but were thought to be too open to interpretation at local levels. Setting and managing 'realistic' expectations of staff was seen as a priority. Staff attitudes towards ERAS tend to become more favourable over time, as practices become successfully 'normalised'. Strengths of the review are that it includes a wide range of different studies, a variety of clinical populations, diversity of methodological approaches and local contexts. Its limitation is the inclusion of a small number of studies, although these represent six countries and four surgical specialties, and so our findings are likely to be transferable. Staff feel positive about the implementation of ERAS, but find the process is complex and challenging. Challenges can be addressed by ensuring that multidisciplinary teams understand ERAS principles and guidelines, and communicate well with one another and with patients. Provision of comprehensive, coherent and locally relevant information to health professionals is helpful. Identifying and recruiting local ERAS champions is likely to improve the implementation and delivery of ERAS pathways. CRD42017059952.


Osteogenic differentiation of late-outgrowth CD45-negative endothelial progenitor cells.
Late-outgrowth CD45-negative endothelial colony-forming cells are implicated to be circulating endothelial progenitor cells (EPCs), as they express endothelial cell markers and can directly form blood vessels. As these cells share characteristics of other progenitor cell phenotypes, late-outgrowth EPCs were assayed for both multilineage differentiation capability and for markers of pluripotency. Clonal single-colony late-outgrowth EPCs were derived from human cord blood and assayed both for multilineage differentiation capability in vitro and for markers of pluripotency by qPCR. Under osteogenic growth conditions, these EPCs expressed the osteogenic markers RUNX2, COL1A1, ALPL, and osteocalcin and demonstrated calcium deposition and bone mineralization. Endothelial colony-forming cells expressed markers associated with induced pluripotent stem cells, including SOX2, POU5F1, c-MYC, and KLF4. Late-outgrowth EPCs express markers associated with pluripotency and can directly express an osteogenic phenotype under bone differentiation conditions in vitro.


Fibroblastic reticular cell-derived lysophosphatidic acid regulates confined intranodal T-cell motility.
Lymph nodes (LNs) are highly confined environments with a cell-dense three-dimensional meshwork, in which lymphocyte migration is regulated by intracellular contractile proteins. However, the molecular cues directing intranodal cell migration remain poorly characterized. Here we demonstrate that lysophosphatidic acid (LPA) produced by LN fibroblastic reticular cells (FRCs) acts locally to LPA2 to induce T-cell motility. In vivo, either specific ablation of LPA-producing ectoenzyme autotaxin in FRCs or LPA2 deficiency in T cells markedly decreased intranodal T cell motility, and FRC-derived LPA critically affected the LPA2-dependent T-cell motility. In vitro, LPA activated the small GTPase RhoA in T cells and limited T-cell adhesion to the underlying substrate via LPA2. The LPA-LPA2 axis also enhanced T-cell migration through narrow pores in a three-dimensional environment, in a ROCK-myosin II-dependent manner. These results strongly suggest that FRC-derived LPA serves as a cell-extrinsic factor that optimizes T-cell movement through the densely packed LN reticular network.


A framework for examining the function of digital health technologies for weight management.
Research is rapidly extending its focus to develop and evaluate weight management interventions that incorporate eHealth technologies. Comparative effectiveness of eHealth interventions is partly limited by the extensive heterogeneity in intervention design, variation in use of eHealth tools, and expanding development of novel tools to promote weight management. We closely examined, characterized, and categorized the use and function of eHealth tools across a wide range of eHealth interventions for weight management in order to first create a novel schematic framework for eHealth interventions and, second, to evaluate eHealth interventions using this framework. We examined 49 randomized controlled trials from two systematic reviews evaluating the effectiveness of eHealth interventions for weight loss. Further characterization of each intervention identified common use and function of eHealth tools represented within interventions and thus important to include in the proposed framework. This resulted in six descriptive domains. We then categorized each eHealth intervention within the context of the newly developed framework. Last, we examined efficacious interventions in the context of the framework. Twenty-five randomized controlled trials reported significantly more weight loss between the intervention group utilizing eHealth, compared to a non-eHealth control intervention and/or within an eHealth intervention group. Of these 25 interventions, 15 (60%) used automated feedback (Domain 1), 13 (52%) used non-eHealth tailored feedback by a health care provider (Domain 5), and 8 (32%) used tailored feedback from a health care professional through an electronic channel (Domain 2). The proposed schematic framework offers an alternative and novel approach for comparing across interventions and informing the development and evaluation of eHealth interventions.


Imagery as a Method of Enhancing Transfer From Trailing to Performance.
This study investigated the effect of engaging in imagery in conjunction with nonspecific training on the transfer of the training to performance. Forty subjects were pretested on a Wingate cycle ergometer test for peak power and a 40-m sprint. Subjects were assigned to one of four groups: imagery training (IT), power training (PT), imagery and power training (DPT), and control (C). Following a 6-week training period, all subjects were retested. Although a MANOVA revealed no significant difference between groups on any variable, the groups-by-time interaction was significant. Therefore an analysis of difference scores on both tests was performed. This analysis revealed that although both the IPT and the PT group significantly improved in peak power, only the IPT group improved significantly on the sprint. The results indicate that imagery coupled with nonspecific training contributes to the enhancement of subsequent performance significantly better than does nonspecific training alone.


Exposure to general anesthesia in early life and the risk of attention deficit/hyperactivity disorder development: a nationwide, retrospective matched-cohort study.
Postnatal exposure to anesthetics can cause neural apoptosis and degeneration in animals, but results from studies conducted on humans were discordant. Previous studies contained no information on the relationship between neurobehavioural disorders and anesthesia exposure in Asian children. We conducted a retrospective matched-cohort study in Taiwan to investigate the association of early life anesthesia exposure with risk of attention deficit/hyperactivity disorder (ADHD). Data were obtained from the National Health Insurance Research Database of Taiwan. Children born between January 1, 2001 and December 31, 2005 were included. Each child with anesthesia exposure before 3 years of age was matched to four unexposed children. Observation was concluded on December 31, 2010. Proportional hazards regression was used to assess the association of anesthesia exposure with ADHD. Analyses were also made based on exposure number and age at the time of first exposure. This matched-cohort comprised of 16 465 children, among which 3293 were exposed to general anesthesia before age 3 years. The adjusted hazard ratio of developing ADHD was 1.06 (95% CI: 0.86,1.31) for general anesthesia exposure. The adjusted hazard ratio of developing ADHD for single and multiple exposures were 1.11 (95% CI: 0.88, 1.41) and 0.96 (95% CI: 0.71,1.31), respectively. No trend of increasing risk was noted based on age at the time of first exposure. Exposure to general anesthesia before 3 years of age was not associated with ADHD.


Burden of Liver Disease among Community-Based People Who Inject Drugs (PWID) in Chennai, India.
We characterize the burden of liver disease in a cohort of PWID in Chennai, India, with a high prevalence of HCV. 1,042 PWID were sampled through community outreach in Chennai. Participants underwent fasting blood draw, questionnaire and an examination that included liver stiffness assessment using transient elastography (Fibroscan) and assessment of steatosis via ultrasound. The median age was 39 years, all were male, 14.8% were HIV infected and 35.6% were HCV antibody positive, of whom 78.9% were chronically infected (HCV RNA positive). Median liver stiffness was 6.2 kPA; 72.9% had no evidence of or mild stiffness, 14.5% had moderate stiffness, and 12.6% had evidence of severe stiffness/cirrhosis. Prevalence of severe stiffness/cirrhosis was significantly higher among persons who were older, had a longer duration of injecting drugs, higher body mass index, higher prevalence of insulin resistance, higher prevalence of steatosis, higher HCV RNA levels and evidence of alcohol dependence. An estimated 42.1% of severe stiffness/cirrhosis in this sample was attributable to HCV. 529 (53.0%) had some evidence of steatosis. Prevalence of steatosis was higher among those who had larger waist circumference, insulin resistance, higher HDL cholesterol and a history of antiretroviral therapy. We observed a high burden of liver disease in this relatively young cohort that was primarily driven by chronic HCV infection, metabolic factors (insulin resistance and steatosis) and heavy alcohol use. Interventions to improve access to HCV treatment and reduce alcohol use are needed to prevent further progression of liver disease.


PHYSIOLOGY. Regulation of breathing by CO₂ requires the proton-activated receptor GPR4 in retrotrapezoid nucleus neurons.
Blood gas and tissue pH regulation depend on the ability of the brain to sense CO2 and/or H(+) and alter breathing appropriately, a homeostatic process called central respiratory chemosensitivity. We show that selective expression of the proton-activated receptor GPR4 in chemosensory neurons of the mouse retrotrapezoid nucleus (RTN) is required for CO2-stimulated breathing. Genetic deletion of GPR4 disrupted acidosis-dependent activation of RTN neurons, increased apnea frequency, and blunted ventilatory responses to CO2. Reintroduction of GPR4 into RTN neurons restored CO2-dependent RTN neuronal activation and rescued the ventilatory phenotype. Additional elimination of TASK-2 (K(2P)5), a pH-sensitive K(+) channel expressed in RTN neurons, essentially abolished the ventilatory response to CO2. The data identify GPR4 and TASK-2 as distinct, parallel, and essential central mediators of respiratory chemosensitivity.


Elimination of two reef-building hydrocorals following the 1982-83 el nino warming event.
One probable extinction and one range reduction of eastern Pacific reef-building hydrocoral (Millepora) species mark the first documented cases of species eliminations resulting from the worldwide 1980s coral reef bleaching events. Two of 12 Panamanian coral species were eliminated suddenly from their former ranges by prolonged high sea temperatures during the 1982-83 El Niño-Southern Oscillation event. Three conditions contributed to their demise: high sensitivity to sea warming, populations confined to a small geographic area, and bathymetric restriction to the euphotic zone (</=20 meters depth) where El Niño sea warming had its greatest effect.


Rod microglia: elongation, alignment, and coupling to form trains across the somatosensory cortex after experimental diffuse brain injury.
Since their discovery, the morphology of microglia has been interpreted to mirror their function, with ramified microglia constantly surveying the micro-environment and rapidly activating when changes occur. In 1899, Franz Nissl discovered what we now recognize as a distinct microglial activation state, microglial rod cells (Stäbchenzellen), which he observed adjacent to neurons. These rod-shaped microglia are typically found in human autopsy cases of paralysis of the insane, a disease of the pre-penicillin era, and best known today from HIV-1-infected brains. Microglial rod cells have been implicated in cortical 'synaptic stripping' but their exact role has remained unclear. This is due at least in part to a scarcity of experimental models. Now we have noted these rod microglia after experimental diffuse brain injury in brain regions that have an associated sensory sensitivity. Here, we describe the time course, location, and surrounding architecture associated with rod microglia following experimental diffuse traumatic brain injury (TBI). Rats were subjected to a moderate midline fluid percussion injury (mFPI), which resulted in transient suppression of their righting reflex (6 to 10 min). Multiple immunohistochemistry protocols targeting microglia with Iba1 and other known microglia markers were undertaken to identify the morphological activation of microglia. Additionally, labeling with Iba1 and cell markers for neurons and astrocytes identified the architecture that surrounds these rod cells. We identified an abundance of Iba1-positive microglia with rod morphology in the primary sensory barrel fields (S1BF). Although present for at least 4 weeks post mFPI, they developed over the first week, peaking at 7 days post-injury. In the absence of contusion, Iba1-positive microglia appear to elongate with their processes extending from the apical and basal ends. These cells then abut one another and lay adjacent to cytoarchitecture of dendrites and axons, with no alignment with astrocytes and oligodendrocytes. Iba1-positive rod microglial cells differentially express other known markers for reactive microglia including OX-6 and CD68. Diffuse traumatic brain injury induces a distinct rod microglia morphology, unique phenotype, and novel association between cells; these observations entice further investigation for impact on neurological outcome.


What are frail older people prepared to endure to achieve improved mobility following hip fracture? A Discrete Choice Experiment.
To investigate the preferences of frail older people for individualised multidisciplinary rehabilitation to promote recovery from a hip fracture. Discrete Choice Experiment. Acute and Rehabilitation Hospitals in Adelaide, South Australia. Eighty-seven patients with recent hip fracture (16 living in residential care facilities prior to fracture). Patients providing informed consent (or consenting family carer proxies in cases where patients were unable to provide informed consent (n = 10)) participated in a face to face interview following surgery to repair a fractured hip to assess their preferences for different configurations of rehabilitation programs. Overall, participants expressed a strong preference for improvements in mobility and a willingness to participate in rehabilitation programs involving moderate pain and effort. However, negative preferences were observed for extremely painful interventions involving high levels of effort (2 h per day for 2 months). Subgroup analysis revealed consistently similar preferences according to place of residence (residential care vs community). Improvements in mobility are highly valued by frail older people recovering from hip fracture, including those living in residential care. Further research should be directed towards achieving greater equity in access to rehabilitation services for the wide spectrum of patients attending hospital with hip fractures.


A Systems Biology Approach to Deciphering the Etiology of Steatosis Employing Patient-Derived Dermal Fibroblasts and iPS Cells.
Non-alcoholic fatty liver disease comprises a broad spectrum of disease states ranging from simple steatosis to non-alcoholic steatohepatitis. As a result of increases in the prevalences of obesity, insulin resistance, and hyperlipidemia, the number of people with hepatic steatosis continues to increase. Differences in susceptibility to steatohepatitis and its progression to cirrhosis have been attributed to a complex interplay of genetic and external factors all addressing the intracellular network. Increase in sugar or refined carbohydrate consumption results in an increase of insulin and insulin resistance that can lead to the accumulation of fat in the liver. Here we demonstrate how a multidisciplinary approach encompassing cellular reprogramming, transcriptomics, proteomics, metabolomics, modeling, network reconstruction, and data management can be employed to unveil the mechanisms underlying the progression of steatosis. Proteomics revealed reduced AKT/mTOR signaling in fibroblasts derived from steatosis patients and further establishes that the insulin-resistant phenotype is present not only in insulin-metabolizing central organs, e.g., the liver, but is also manifested in skin fibroblasts. Transcriptome data enabled the generation of a regulatory network based on the transcription factor SREBF1, linked to a metabolic network of glycerolipid, and fatty acid biosynthesis including the downstream transcriptional targets of SREBF1 which include LIPIN1 (LPIN) and low density lipoprotein receptor. Glutathione metabolism was among the pathways enriched in steatosis patients in comparison to healthy controls. By using a model of the glutathione pathway we predict a significant increase in the flux through glutathione synthesis as both gamma-glutamylcysteine synthetase and glutathione synthetase have an increased flux. We anticipate that a larger cohort of patients and matched controls will confirm our preliminary findings presented here.


Curcumin and turmeric delay streptozotocin-induced diabetic cataract in rats.
The purpose of this study was to investigate the effect of curcumin and its source, turmeric, on streptozotocin-induced diabetic cataract in rats. Wistar-NIN rats were selected and diabetes was induced by streptozotocin (35 mg/kg body weight, intraperitoneally) and divided into four groups (group II-V). The control (group I) rats received only vehicle. Group I and II animals received an unsupplemented AIN-93 diet, and those in groups III, IV, and V received 0.002% and 0.01% curcumin and 0.5% turmeric, respectively, in an AIN-93 diet for a period of 8 weeks. Cataract progression due to hyperglycemia was monitored by slit lamp biomicroscope and classified into four stages. At the end of 8 weeks, the animals were killed and the biochemical pathways involved in the pathogenesis of cataract such as oxidative stress, polyol pathway, alterations in protein content and crystallin profile in the lens were investigated, to understand the possible mechanism of action of curcumin and turmeric. Blood glucose and insulin levels were also determined. Although, both curcumin and turmeric did not prevent streptozotocin-induced hyperglycemia, as assessed by blood glucose and insulin levels, slit lamp microscope observations indicated that these supplements delayed the progression and maturation of cataract. The present studies suggest that curcumin and turmeric treatment appear to have countered the hyperglycemia-induced oxidative stress, because there was a reversal of changes with respect to lipid peroxidation, reduced glutathione, protein carbonyl content and activities of antioxidant enzymes in a significant manner. Also, treatment with turmeric or curcumin appears to have minimized osmotic stress, as assessed by polyol pathway enzymes. Most important, aggregation and insolubilization of lens proteins due to hyperglycemia was prevented by turmeric and curcumin. Turmeric was more effective than its corresponding levels of curcumin. The results indicate that turmeric and curcumin are effective against the development of diabetic cataract in rats. Further, these results imply that ingredients in the study's dietary sources, such as turmeric, may be explored for anticataractogenic agents that prevent or delay the development of cataract.


A facile one-step solution deposition via non-solvent/solvent mixture for efficient organometal halide perovskite light-emitting diodes.
Although organometal halide perovskite materials have shown great potential in light-emitting diodes, their performance is greatly restricted by the poor morphology of the perovskite layer. In this work, we demonstrate a facile one-step solution method to improve the perovskite film morphology via a non-solvent/solvent mixture. An efficient CH3NH3PbBr3-based light-emitting diode was prepared with a chlorobenzene/N,N-dimethylformamide mixed solvent. A high efficiency of 0.54 cd A(-1) is demonstrated, which is 22 times higher than that of a device fabricated by a traditional one-step solution process. Furthermore, the uniformity of the emission region and the device stability are strongly improved by this facile one-step solution process. Our work paves a new way for the morphological control of perovskite films for application in light-emitting diodes.


P2Y2 receptor knock-out mice display normal NaCl absorption in medullary thick ascending limb.
Local purinergic signals modulate renal tubular transport. Acute activation of renal epithelial P2 receptors causes inhibition of epithelial transport and thus, should favor increased water and salt excretion by the kidney. So far only a few studies have addressed the effects of extracellular nucleotides on ion transport in the thick ascending limb (TAL). In the medullary thick ascending limb (mTAL), basolateral P2X receptors markedly (~25%) inhibit NaCl absorption. Although this segment does express both apical and basolateral P2Y2 receptors, acute activation of the basolateral P2Y2 receptors had no apparent effect on transepithelial ion transport. Here we studied, if the absence of the P2Y2 receptor causes chronic alterations in mTAL NaCl absorption by comparing basal and AVP-stimulated transepithelial transport rates. We used perfused mouse mTALs to electrically measure NaCl absorption in juvenile (<35 days) and adult (>35 days) male mice. Using microelectrodes, we determined the transepithelial voltage (Vte) and the transepithelial resistance (Rte) and thus, transepithelial NaCl absorption (equivalent short circuit current, I'sc). We find that mTALs from adult wild type (WT) mice have significantly lower NaCl absorption rates when compared to mTALs from juvenile WT mice. This could be attributed to significantly higher Rtevalues in mTALs from adult WT mice. This pattern was not observed in mTALs from P2Y2 receptor knockout (KO) mice. In addition, adult P2Y2 receptor KO mTALs have significantly lower Vtevalues compared to the juvenile. No difference in absolute I'sc was observed when comparing mTALs from WT and KO mice. AVP stimulated the mTALs to similar increases of NaCl absorption irrespective of the absence of the P2Y2 receptor. No difference was observed in the medullary expression level of NKCC2 in between the genotypes. These data indicate that the lack of P2Y2 receptors does not cause substantial differences in resting and AVP-stimulated NaCl absorption in mouse mTAL.


The effects of tension on acetylstrophanthidin-induced transient depolarizations and aftercontractions in canine myocardial and Purkinje tissues.
Transmembrane potentials and contractile activity were recorded from isolated canine Purkinje and ventricular muscle preparations exposed to acetylstrophantidin (AS) and subjected to a resting tension equal to 80% of that required to elicit peak developed tension. AS induced transient depolarizations (TD's) accompanied by aftercontractions in Purkinje tissue. AS also induced aftercontractions in 11 muscle preparations, and in seven of these the mechanical events were associated with TD's. Aftercontractions and TD's in both and the coupling intervals were directly related to the preceding basic cycle length (BCL). The amplitudes of aftercontractions and TD's reached a maximum at a coupling interval of 600-700 msec. Tension increased the amplitude of TD's in Purkinje tissue and promoted the appearance of TD's in muscle. TD's in muscle occasionally reached threshold in the presence of tension. The results of this study suggest that stretch or increased resting tension may promote the types of cardiac arrhythmias that are causally related to digitalis-induced TD's. The results are compatible with the hypothesis that TD's are caused either by a transmembrane influx of calcium or by an internal release of calcium ions.


Brief Mindfulness Meditation for Depression and Anxiety Symptoms in Patients Undergoing Hemodialysis: A Pilot Feasibility Study.
Up to 50% of patients undergoing hemodialysis suffer from symptoms of depression and/or anxiety. Access to traditional pharmacotherapies and psychotherapies for depression or anxiety in this patient population has been inadequate. The objective of this study was to investigate the feasibility and effectiveness of brief mindfulness meditation intervention for patients on hemodialysis with depression and anxiety symptoms. This study was a randomized, controlled, assessor-blinded trial conducted in an urban hemodialysis unit. Forty-one patients were randomly assigned to intervention (n=21) and treatment-as-usual (n=20) groups. The intervention group received an 8-week individual chairside meditation intervention lasting 10-15 minutes, three times a week during hemodialysis. Feasibility outcomes were primarily assessed: enrollment rates, intervention completion rates, and intervention tolerability. Symptoms of depression and anxiety were measured using the Patient Health Questionnaire (PHQ-9) and the General Anxiety Disorder-7 (GAD-7). Of those deemed eligible for the study, 67% enrolled (41 of 61). Of the participants randomized to the intervention group, 71% completed the study, with meditation being well tolerated (median rating of 8 of 10 in a Likert scale; interquartile range=10-5 of 10). Barriers to intervention delivery included frequent hemodialysis shift changes, interruptions by staff or alarms, space constraints, fluctuating participant medical status, and participant fatigue. Meditation was associated with subjective benefits but no statistically significant effect on depression scores (change in PHQ-9, -3.0±3.9 in the intervention group versus -2.0±4.7 in controls; P=0.45) or anxiety scores (change in GAD-7, -0.9±4.6 versus -0.8±4.8; P=0.91). On the basis of the results of this study, mindfulness meditation appears to be feasible and well tolerated in patients on hemodialysis with anxiety and depression symptoms. The study did not reveal significant effects of the interventions on depression and anxiety scores. This article contains a podcast at https://www.asn-online.org/media/podcast/CJASN/2017_10_12_CJASNPodcast_17_12_.mp3.


Decreased blood vessel density and endothelial cell subset dynamics during ageing of the endocrine system.
Age-associated alterations of the hormone-secreting endocrine system cause organ dysfunction and disease states. However, the cell biology of endocrine tissue ageing remains poorly understood. Here, we perform comparative 3D imaging to understand age-related perturbations of the endothelial cell (EC) compartment in endocrine glands. Datasets of a wide range of markers highlight a decline in capillary and artery numbers, but not of perivascular cells in pancreas, testis and thyroid gland, with age in mice and humans. Further, angiogenesis and β-cell expansion in the pancreas are coupled by a distinct age-dependent subset of ECs. While this EC subpopulation supports pancreatic β cells, it declines during ageing concomitant with increased expression of the gap junction protein Gja1. EC-specific ablation of Gja1 restores β-cell expansion in the aged pancreas. These results provide a proof of concept for understanding age-related vascular changes and imply that therapeutic targeting of blood vessels may restore aged endocrine tissue function. This comprehensive data atlas offers over > 1,000 multicolour volumes for exploration and research in endocrinology, ageing, matrix and vascular biology.


Structure of the oncoprotein Rcl bound to three nucleotide analogues.
Rcl is a novel N-glycoside hydrolase found in mammals that shows specificity for the hydrolysis of 5'-monophosphate nucleotides. Its role in nucleotide catabolism and the resulting production of 2-deoxyribose 5-phosphate has suggested that it might fuel cancer growth. Its expression is regulated by c-Myc, but its role as an oncoprotein remains to be clarified. In parallel, various nucleosides have been shown to acquire pro-apoptotic properties upon 5'-monophosphorylation in cells. These include triciribine, a tricyclic nucleoside analogue that is currently in clinical trials in combination with a farnesyltransferase inhibitor. Similarly, an N(6)-alkyl-AMP has been shown to be cytotoxic. Interestingly, Rcl has been shown to be inhibited by such compounds in vitro. In order to gain better insight into the precise ligand-recognition determinants, the crystallization of Rcl with these nucleotide analogues was attempted. The first crystal structure of Rcl was solved by molecular replacement using its NMR structure in combination with distantly related crystal structures. The structures of Rcl bound to two other nucleotides were then solved by molecular replacement using the previous crystal structure as a template. The resulting structures, solved at high resolution, led to a clear characterization of the protein-ligand interactions that will guide further rational drug design.