BCL2 oncogene protein expression in human hematopoietic precursors during fetal life.
BCL2 proto-oncogene encodes a 25-kD protein that is characteristically localized in the inner mitochondrial membrane of the cell. It has been reported that BCL2 protein has the unique functional role of blocking programmed cells death without affecting proliferation. We have analyzed the expression of the BCL2 protein in fetal hematopoietic tissues from the 10th week of gestational age onward. Fetal thymus, liver, and bone marrow and cord blood were investigated. The experiments were performed by the alkaline-antialkaline phosphatase (APAAP) technique by staining air-dried acetone-fixed cytospins and by dual-color immunofluorescent assay by staining mononuclear cell suspensions with monoclonal antibodies detecting BCL2 protein and antigens expressed by different hematopoietic subsets. Flow cytometric analyses were performed on FACSort's Comsort 32 (Becton Dickinson, San Jose, CA). The results have shown that the BCL2 protein is expressed in human fetal ontogenesis at the earliest stages examined. The major conceptual aspects of the results are 1) BCL2 is largely expressed in the hematopoietic cells during ontogenesis. BCL2+ cells include both immature and more differentiated subsets. Moreover, the 25-kD protein is expression in cell subsets well known to be high proliferating. This behavior suggests that BCL2 could have more complex functions than those previously described. 2) The expression in the major part of CD34+ cells suggests that BCL2 could play a role in stem cell survival. 3) BCL2 is expressed in not only medullary but also cortical thymocytes, where it could cooperate in positive selection processes. 4) The involvement of BCL2 in the immunosurveillance is indicated not only by its role in B and T cell lineages but also by its expression in particular subsets like that of the cytoplasmic CD3+ fetal liver NK cells. 5) The discrepancy observed between the results of transgenic mice analysis and in vitro inhibition experiments by antisense oligonucleotides performed for understanding BCL2 functions must stress the importance of the direct immunologic analysis of BCL2 in human hematopoietic cells.


Current strategies in the management of hypereosinophilic syndrome, including mepolizumab.
Patients with hypereosinophilic syndrome (HES) vary considerably in their clinical presentation with regard to the severity and pattern of end-organ involvement. Clinical manifestations range from nonspecific symptoms to life-threatening, multisystem damage caused by eosinophil infiltration and local release of proinflammatory mediators and toxic granule products from these invading cells. The primary objective of treatment is to reduce blood and tissue eosinophilia and prevent eosinophil-mediated tissue damage as safely as possible. Systemic corticosteroids, such as prednisone, are first-line therapy for the management of patients with symptomatic HES who lack the Fip1-like 1-platelet-derived growth factor receptor-alpha (FIP1L1-PDGFRA) gene fusion mutation. The tyrosine kinase inhibitor, imatinib, is first-line treatment for FIP1L1-PDGFRA-positive patients). Because of the toxicity and serious side-effects that can occur with oral corticosteroids, alternative therapies may need to be introduced to reduce the cumulative corticosteroid exposure while maintaining disease control. Among corticosteroid-sparing agents are cytotoxic drugs and interferon-alpha; anti-interleukin-5 (IL-5) monoclonal antibodies are also currently under investigation for the treatment of HES. This manuscript reviews the available treatments for HES and the range of side-effects associated with long-term corticosteroid use, and then focuses on the anti-IL-5 monoclonal antibodies, mepolizumab and reslizumab. Of these, only mepolizumab has been studied in a randomized, placebo-controlled trial. Literature search methodology utilized www.pubmed.gov and www.clinicaltrials.gov with search terms including hypereosinophilic syndrome and corticosteroid side-effects coupled with search terms including eosinophils, mepolizumab and reslizumab through March 2010. Three case studies are presented that demonstrate the limitations of corticosteroid therapy in terms of tolerability and quality of life, and the subsequent use of mepolizumab as a corticosteroid-sparing agent in these individuals. Targeted eosinophil-directed therapy with an anti-IL-5 neutralizing monoclonal antibody reduced the need for corticosteroids in these three HES patients without disease exacerbations.


[Remaxol hepatoprotective therapy of patients with tuberculosis and HIV infection in day unit of tuberculosis dispensary].
To improve the treatment of hepatotoxic responses to antituberculosis polychemotherapy, the impact of remaxol on the biochemical indices and parameters of the antioxidant system in patients with tuberculosis and HIV infection was estimated. The use of remaxol having cytoprotective, anticholestatic, antihypoxitic and antioxidant effects in the treatment of patients with tuberculosis and HIV infection and liver drug damage due to tuberculosis polychemotherapy significantly improved the biochemical indices and lowered the level of the cytolytic and cholestatic syndromes. Remaxol increased the antioxidant system potential and had an antihypoxitic effect.


NBR1 acts as an autophagy receptor for peroxisomes.
Selective macro-autophagy is an intracellular process by which large cytoplasmic materials are selectively sequestered and degraded in the lysosomes. Substrate selection is mediated by ubiquitylation and recruitment of ubiquitin-binding autophagic receptors such as p62, NBR1, NDP52 and Optineurin. Although it has been shown that these receptors act cooperatively to target some types of substrates to nascent autophagosomes, their precise roles are not well understood. We examined selective autophagic degradation of peroxisomes (pexophagy), and found that NBR1 is necessary and sufficient for pexophagy. Mutagenesis studies of NBR1 showed that the amphipathic α-helical J domain, the ubiquitin-associated (UBA) domain, the LC3-interacting region and the coiled-coil domain are necessary to mediate pexophagy. Strikingly, substrate selectivity is partly achieved by NBR1 itself by coincident binding of the J and UBA domains to peroxisomes. Although p62 is not required when NBR1 is in excess, its binding to NBR1 increases the efficiency of NBR1-mediated pexophagy. Together, these results suggest that NBR1 is the specific autophagy receptor for pexophagy.


[Hemodynamic effect of intra-aortic balloon counterpulsation in the acute period of myocardial infarct].
The author studied changes in central hemodynamics during counterpulsation with an intra-aortic balloon in 38 patients in the acute period of myocardial infarction. The main hemodynamic values were determined by thermodilution. A positive effect of counterpulsation on the cardiac output was revealed in patients with congestive cardiac insufficiency and marked disorders of rhythm and conduction. It is recommended that counterpulsation with an intra-aortic balloon be included in the complex of measures for treating acute hemodynamic disorders in the first 24 hours of complicated myocardial infarction.


Conditional targeting of Ispd using paired Cas9 nickase and a single DNA template in mice.
CRISPR/Cas9 technology is a highly promising genome editing tool in the mouse, potentially overcoming the costs and time required for more traditional gene targeting methods in embryonic stem (ES) cells. Recently, compared to the wildtype nuclease, paired Cas9 nickase (Cas9n) combined with single guide RNA (sgRNA) molecules has been found to enhance the specificity of genome editing while reducing off-target effects. Paired Cas9n has been shown to be as efficient as Cas9 for generating insertion and deletion (indel) mutations by non-homologous end joining and targeted deletion in the genome. However, an efficient and reliable approach to the insertion of loxP sites flanking critical exon(s) to create a conditional allele of a target gene remains an elusive goal. In this study, we microinjected Cas9n RNA with sgRNAs together with a single DNA template encoding two loxP sites flanking (floxing) exon 2 of the isoprenoid synthase containing domain (Ispd) into the pronucleus and cytoplasm of C57BL/6NCr one-cell stage zygotes. After surgical transfer, one F0 mouse expressing a conditional allele was produced (at a frequency of ∼8% of live pups born). The floxed allele was transmitted through the germline to F1 progeny, and could be successfully recombined using Cre recombinase. This study indicates that conditional targeting can be accomplished effectively using paired Cas9n and a single DNA template.


Non-pharmacological interventions for improving postpartum maternal sleep: A systematic review and meta-analysis.
Evidence suggests that poor postpartum sleep quality is a risk factor for the development of postpartum depression. As such, non-pharmacological interventions have been developed to help improve sleep in the postpartum period. The primary aims of this systematic review and meta-analysis were to determine if non-pharmacological interventions improved maternal sleep and to compare the effectiveness of different intervention types. Secondary aims included examining effects on maternal mood and infant sleep. We searched MEDLINE, EMBASE, CINAHL, PsycINFO, and Web of Science from their inceptions to September 2017 and found 15 eligible studies. Non-pharmacological sleep interventions were found to improve subjective reports of maternal sleep (Cohen's d = -0.54, 95%CI = -0.88 to -0.19). Massage (Cohen's d = -1.07 95%CI = -1.34 to -0.79) and exercise (Cohen's d = -0.82 95%CI = -1.28 to -0.37) interventions had the largest impact on maternal sleep quality. Positive effects on nocturnal infant sleep were found for interventions overall (Cohen's d = -0.27 95%CI = -0.52 to -0.02) but not for maternal depression (Cohen's d = -0.08 95%CI = -0.28 to 0.12). Despite evidence suggesting improvements in subjective maternal sleep, more research must be conducted on the durability of effects of non-pharmacological interventions using objective measures of sleep quality.


Effects of lysine, threonine and methionine on light-driven protein synthesis in isolated pea (Pisum sativum L.) chloroplasts.
Light-driven incorporation of [(14)C]leucine (LEU) into protein in isolated pea chloroplasts was inhibited by 0.1 and 1 mM lysine (LYS), 0.1-10 mM threonine (THR) and 1 mM methionine (MET). Equimolar combinations of LYS plus THR were inhibitory at both 0.1 and 0.5 mM. Incorporation of [(14)C] aspartic acid (ASP) and [(3)H]tyrosine (TYR) was also reduced by 1 mM LYS or THR. In the cases tested, LYS and/or THR inhibitions were partially or fully reversed by 0.1 mM MET. [(35)S]MET incorporation was unaffected or stimulated by LYS and THR. These data are consistent with the hypothesis that MET is biosynthesized in isolated chloroplasts and that its synthesis is regulated by LYS and/or THR. Of 16 other amino acids tested at 1 mM, isoleucine, phenylalanine, tryptophan, tyrosine and valine inhibited protein synthesis.


Chromosome-level genome assembly of chub mackerel (Scomber japonicus) from the Indo-Pacific Ocean.
Chub mackerels (Scomber japonicus) are a migratory marine fish widely distributed in the Indo-Pacific Ocean. They are globally consumed for their high Omega-3 content, but their population is declining due to global warming. Here, we generated the first chromosome-level genome assembly of chub mackerel (fScoJap1) using the Vertebrate Genomes Project assembly pipeline with PacBio HiFi genomic sequencing and Arima Hi-C chromosome contact data. The final assembly is 828.68 Mb with 24 chromosomes, nearly all containing telomeric repeats at their ends. We annotated 31,656 genes and discovered that approximately 2.19% of the genome contained DNA transposon elements repressed within duplicated genes. Analyzing 5-methylcytosine (5mC) modifications using HiFi reads, we observed open/close chromatin patterns at gene promoters, including the FADS2 gene involved in Omega-3 production. This chromosome-level reference genome provides unprecedented opportunities for advancing our knowledge of chub mackerels in biology, industry, and conservation.


Potential circulating markers for the management of kidney cancer (Review).
The prognosis of renal cell carcinoma (RCC) is generally poor. An easier detection of this tumor and a better monitoring of RCC patients would be possible if serum markers with acceptable sensitivity and specificity were available. In RCC, as opposed to other cancers, no circulating serum markers with sufficient renal specificity have been discovered. In fact, even when the hybridoma technology allowed the production of several monoclonal antibodies against RCC structures, none of them led to any available diagnostic immunoassays. Other possible circulating tumor markers of potential application in RCC patients include different substances such as acute phase reactant proteins, enzymes, mucins, cytokeratins, proteins, interleukins, that demonstrated some relationship with the presence and the changes in the RCC evolution. In this general review we report and discuss the results in the literature obtained by serum assays of these substances which have been shown to be of some help for the prognosis and monitoring of RCC. The greater part of these biomolecules are already measured in clinical practice for the management of other malignancies, but their application in RCC could give interesting clinical information.


Cost effectiveness of donepezil in the treatment of mild to moderate Alzheimer's disease: a UK evaluation using discrete-event simulation.
Recommendations in the UK suggest restricting treatment of Alzheimer's disease with cholinesterase inhibitors, on cost-effectiveness grounds, to patients with moderate cognitive decline. As the economic analyses that informed these recommendations have been the subject of debate, we sought to address the potential limitations of existing models and produce estimates of donepezil treatment cost effectiveness in the UK using the most recent available data and simulation techniques. A discrete-event simulation was developed that predicts progression of Alzheimer's disease through correlated changes in cognition, behavioural disturbance and function. Patient-level data from seven randomized, placebo-controlled donepezil trials and a 7-year follow-up registry provided the basis for modeling longitudinal outcomes. Individuals in the simulation were assigned unique demographic and clinical characteristics and then followed for 10 years, with severity of disease tracked on continuous scales. Patient mix and costs were developed from UK-specific literature. Analyses were run for severity subgroups to evaluate outcomes for sub-populations with disease of mild versus moderate severity from both a healthcare payer and societal perspective. All costs are reported in pound, year 2007 values, and all outcomes are discounted at 3.5% per annum. Over 10 years, treatment of all patients with mild to moderate disease reduces overall direct medical costs by an average of over pound2300 per patient. When unpaid caregiver time is also taken into consideration, savings increase to over pound4700 per patient. Compared with untreated patients, patients receiving donepezil experience a discounted gain in QALYs averaging 0.11, with their caregivers gaining, on average, 0.01 QALYs. For the subset of patients starting treatment with more severe disease, savings are more modest, averaging about pound1600 and pound3750 from healthcare and societal perspectives, respectively. In probabilistic sensitivity analyses, donepezil dominated no treatment between 57% and 62% of replications when only medical costs were considered, and between 74% and 79% of replications when indirect costs were included, with results more favourable for treatment initiation in the mild versus moderate severity stages of the disease. Although the simulation results are not definitive, they suggest that donepezil leads to health benefits and cost savings when used to treat mild to moderately severe Alzheimer's disease in the UK. They also indicate that both benefits and savings may be greatest when treatment is started while patients are still in the mild stages of Alzheimer's disease.


Occupational exposure to endocrine-disrupting compounds and biliary tract cancer among men.
This study investigated the association between cancer of the extrahepatic biliary tract and exposure to endocrine-disrupting compounds. Altogether 183 men with histologically confirmed carcinoma of the extrahepatic biliary tract and 1938 matched controls were interviewed between 1995 and 1997 in the frame of an international multicenter case-control study in six European countries (Denmark, France, Germany, Italy, Spain, and Sweden). Self-reported job descriptions were converted to semiquantitative variables (intensity, probability, and duration of exposure) for 14 endocrine-disrupting compounds. The cases were compared with 1421 population controls and 517 colon adenocarcinoma patients. Odds ratios (OR) and 95% confidence intervals (95% CI) were obtained with unconditional logistic regression and adjusted for age, country, and gallstones. Occupational exposure to endocrine-disrupting compounds resulted in an OR of 1.4 (95% CI 1.0-2.1) with no dose-effect relationship for cumulative exposure (low: OR 1.3, 95% CI 0.6-3.0; medium: OR 1.5, 95% CI 0.8-2.7; high: OR 1.4, 95% CI 0.9-2.4) (only index participants). The elevated risk was restricted to extrahepatic bile ducts and ampulla Vateri (OR 1.7, 95% CI 1.0-2.6). The adjusted OR for cancer of the extrahepatic biliary tract after exposure to polychlorinated biphenyls was 2.8 (95% CI 1.3-5.9, only index participants). The data show some associations between exposure to endocrine-disrupting compounds in the workplace and the risk for cancer of the extrahepatic biliary tract among men, particularly for the extrahepatic bile duct and ampulla of Vater. Polychlorinated biphenyls could possibly be a strong risk factor.


Identifying domains of quality of life in children with cancer undergoing palliative care: A qualitative study with professionals.
The goal of pediatric palliative care (PPC) is to maintain the quality of life (QoL) of children whose lives are threatened. However, there are sparse scientific data on the domains of QoL in this particular context, and no measurement strategies are available. The present study aims to describe the domains of QoL in the context of PPC in oncology, according to the perceptions of professional caregivers. Semistructured interviews were conducted with a random sample of 20 professional caregivers from the Division of Hematology/Oncology at Le Centre Hospitalier Universitaire Sainte-Justine (Montréal, Canada). The caregivers were asked about their perceptions about the QoL of the children they have cared for in this context. The data were analyzed using inductive thematic content analysis. The analysis allowed us to identify seven domains of QoL: "physical comfort," "alleviation of psychological suffering," "fun and the present moment," "sense of control," "feeling valued and appreciated," "feeling that life goes on," and "meaningful social relationships." Caregivers recount the regard that should be accorded to maintaining well-being and a sense of fun, as well as fostering the child's abilities, taking account of the progression of the disease, and to fulfilling his or her needs, especially social ones. Our results also demonstrate that all domains were positively referred to by professional caregivers. The data from our study will lead to better assessment of QoL according to the trajectory of a child with advanced cancer while undergoing PPC.


A systematic review and meta-analysis of studies comparing readmission rates and mortality rates in patients with heart failure.
Heart failure is the leading cause of hospitalization and readmission in many hospitals worldwide. We performed a meta-analysis to evaluate the effectiveness of multidisciplinary heart failure management programs on hospital admission rates. We identified studies through an electronic search and mortality using 8 distinct methods. Eligible studies met the following criteria: (1) randomized controlled clinical trials of adult inpatients hospitalized for heart failure enrolled either at the time of discharge or within 1 week after discharge; (2) heart failure-specific patient education intervention coupled with a postdischarge follow-up assessment; and (3) unplanned readmission reported. Four reviewers independently assessed each study for eligibility and quality, achieving a weighted kappa of 0.73 for eligibility and 0.77 for quality. For each study we calculated the relative risk for readmissions and mortality for patients receiving enhanced education relative to patients receiving usual care. A total of 529 citation titles were identified, of which 8 randomized trials proved eligible. The pooled relative risk for hospital readmission rates using a random-effects model was 0.79 (95% confidence interval, 0.68-0.91; P<.001; heterogeneity P = .25). There was no apparent effect on mortality (relative risk, 0.98; 95% confidence interval, 0.72-1.34; P = .90; heterogeneity P = .20). Data were insufficient to meaningfully pool intervention effects on quality of life or compliance. This systematic review suggests that specific heart failure-targeted interventions significantly decrease hospital readmissions but do not affect mortality rates.


Fermented Soybean Suppresses Visceral Fat Accumulation in Mice.
Mice are fed a soybean diet before or after fermentation in the present work to allow direct comparison of the antiobesity effect of fermentation. C57BL6J mice were fed high-fat diets containing boiled soybeans (prefermentation) or Natto (postfermentation) for 4 weeks. Prefermented cooked soybeans or Natto was added at levels of 2.5 and 5%, which are the amounts that can be ingested in a normal diet once and twice a day. In addition, mice gut microbiota from fecal samples were analyzed to explore the mechanisms of effects caused by fermentation. Natto intake significantly reduced visceral fat in a dose-dependent manner, inhibited hypertrophy of adipocytes, improved carbohydrate metabolism, and reduced oxidative stress. These effects were seen in mice fed soybeans before fermentation, but were stronger in mice fed Natto. Therefore, soybean intake has beneficial effects and fermentation of soybeans enhances these effects. Natto was found to suppress fatty acid synthesis and promote fatty acid catabolism in the liver. These effects were also stronger with Natto compared with soybeans before fermentation. In addition, Natto had more potent beneficial effects on gut microbiota compared to soybeans. These results suggest that Natto intake supports maintenance of health.


Competing four-wave mixing processes in dispersion oscillating telecom fiber.
We experimentally study the dynamics of the generation of multiple sidebands by means of a quasi-phase-matched four-wave mixing (FWM) process occurring in a dispersion-oscillating, highly nonlinear optical fiber. The fiber under test is pumped by a ns microchip laser operating in the normal average group-velocity dispersion regime and in the telecom C band. We reveal that the growth of higher-order sidebands is strongly influenced by the competition with cascade FWM between the pump and the first-order quasi-phase matched sidebands. The properties of these competing FWM processes are substantially affected when a partially coherent pump source is used, leading to a drastic reduction of the average power needed for sideband generation.


Validation and investigation of reactive species yields of Geant4-DNA chemistry models.
Indirect biological damage due to reactive species produced in water radiolysis reactions is responsible for the majority of biological effect for low linear energy transfer (LET) radiation. Modeling water radiolysis and the subsequent interactions of reactive species, as well as track structures, is essential to model radiobiology on the microscale. Recently, chemistry models have been developed for Geant4-DNA to be used in combination with the comprehensive existing physics models. In the current work, the first detailed, independent, in silico validation of all species yields with published experimental observations and comparison with other radiobiological simulations is presented. Additionally, the effect of LET of protons and heavier ions on reactive species yield in the model was examined, as well as the completeness of the chemical reactions following the radiolysis within the time after physical interactions simulated in the model. Yields over time of reactive species were simulated for water radiolysis by incident electrons, protons, alpha particles, and ions with various LETs using Geant4 and RITRACKS simulation tools. Water dissociation and recombination was simulated using Geant4 to determine the completeness of chemical reactions at the end of the simulation. Yield validation was performed by comparing yields simulated using Geant4 with experimental observations and other simulations. Validation was performed for all species for low LET radiation and the solvated electron and hydroxyl radical for high LET ions. It was found that the Geant4-DNA chemistry yields were generally in good agreement with experimental observations and other simulations. However, the Geant4-DNA yields for the hydroxyl radical and hydrogen peroxide at the end of the chemistry stage were found to be respectively considerably higher and lower than the experimentally observed yields. Increasing the LET of incident hadrons increased the yield of secondary species and decreased the yield of primary species. The effect of LET on the yield of the hydroxyl radical at 100 ns simulated with Geant4 was in good agreement with experimental measurements. Additionally, by the end of the simulation only 40% of dissociated water molecules had been recombined and the rate of recombination was slowing. The yields simulated using Geant4 are within reasonable agreement with experimental observations. Higher LET radiation corresponds with increased yields of secondary species and decreased yields of primary species. These trends combined with the LET having similar effects on the 100 ns hydroxyl radical yield for Geant4 and experimental measurements indicate that Geant4 accurately models the effect of LET on radiolysis yields. The limited recombination within the modeled chemistry stage and the slowing rate of recombination at the end of the stage indicate potential long-range indirect biological damage.


The role of melatonin in pancreatic protection: could melatonin be used in the treatment of acute pancreatitis?
Acute pancreatitis is a disease, which could be manifested as either a mild edematous form or a more severe necrotizing pancreatitis which has a poor prognosis. The etiology and pathogenesis of this ailment is not completely clear. Melatonin is an indoleamine which is produced from L-tryptophan in the pineal gland and in the other tissue including gastrointestinal tract. Both melatonin and its precursor have been demonstrated to protect the pancreas against acute pancreatitis and to attenuate pancreatic tissue damage. In the pancreas melatonin and L-tryptophan activate complex mechanisms which involve direct scavenging of the radical oxygen and nitrogen species, activation of antioxidant enzymes (catalase, superoxide dysmutase, glutation peroxidase), reduction of pro-inflammatory cytokines and prostaglandins, activation of heat shock protein, and a decrease of necrosis and increase of regeneration in the pancreas. There are several arguments for the idea that endogenous melatonin produced in the pineal gland and in the gastrointestinal system could be the part of a native mechanisms for protecting the pancreas against acute damage: 1/ the melatonin precursor L-tryptophan exerts similar protective effect as melatonin, 2/ application of the melatonin receptor antagonist, luzindole aggravates acute pancreatitis, 3/ pinealectomy results in the exacerbation of acute pancreatitis, 4/ low melatonin plasma levels are associated with an increased risk of severe acute pancreatitis. These observations leads to the idea that perhaps melatonin could be used in clinical trials as supportive therapy in acute pancreatitis.


Contribution of the cortico-claustral loop to receptive field properties in area 17 of the cat.
The contribution of the cat's claustrum to the response properties of cells in area 17 was studied by destroying the left claustrum and examining receptive field properties of cells in both the left and right area 17. For each cell we assessed sharpness of orientation tuning, degree of direction selectivity, length summation, end-stopping, ocular dominance, responsiveness, and several other properties. On the lesioned side, 462 cells were studied, and on the control side, 636 cells. All cats showed a reduction in the number of end-stopped cells in area 17 on the side of the lesion. This was particularly marked in layers 2 + 3 and 4, where end-stopped cells are normally most abundant. In the control hemisphere, 43% of the sample from these layers showed at least moderate end-stopping whereas, in contrast, only 21% exhibited this degree of end-stopping on the side of the lesion. There was no obvious change in other response properties. We conclude that one function of the cat's claustrum is to help regulate the length selectivity of cells in area 17.


The Grb10/Nedd4 complex regulates ligand-induced ubiquitination and stability of the insulin-like growth factor I receptor.
The adapter protein Grb10 belongs to a superfamily of related proteins, including Grb7, -10, and -14 and Caenorhabditis elegans Mig10. Grb10 is an interacting partner of the insulin-like growth factor I receptor (IGF-IR) and the insulin receptor (IR). Previous work showed an inhibitory effect of mouse Grb10 (mGrb10alpha) on IGF-I-mediated mitogenesis (A. Morrione et al., J. Biol. Chem. 272:26382-26387, 1997). With mGrb10alpha as bait in a yeast two-hybrid screen, mouse Nedd4 (mNedd4-1), a ubiquitin protein ligase, was previously isolated as an interacting protein of Grb10 (A. Morrione et al., J. Biol. Chem. 274:24094-24099, 1999). However, Grb10 is not ubiquitinated by Nedd4 in cells. Here we show that in mouse embryo fibroblasts overexpressing Grb10 and the IGF-IR (p6/Grb10), there is a strong ligand-dependent increase in ubiquitination of the IGF-IR compared with that in parental cells (p6). This increased ubiquitination is associated with a shorter half-life and increased internalization of the IGF-IR. The IGF-IR is stabilized following treatment with both MG132 and chloroquine, indicating that both the proteasome and lysosomal pathways mediate degradation of the receptor. Ubiquitination of the IGF-IR likely occurs at the plasma membrane, prior to the formation of endocytic vesicles, as it is insensitive to dansylcadaverine, an inhibitor of early endosome formation in IGF-IR endocytosis. Grb10 coimmunoprecipitates with the IGF-IR and endogenous Nedd4 in p6/Grb10 cells, suggesting the presence of a Grb10/Nedd4/IGF-IR complex. Ubiquitination of the IGF-IR in p6/Grb10 cells is severely impaired by overexpression of a catalytically inactive Nedd4 mutant (Nedd4-CS), which also stabilizes the receptor. Likewise, overexpression of a Grb10 mutant lacking the Src homology 2 (SH2) domain impaired ubiquitination of the IGF-IR in parental p6 and p6/Grb10 cells, indicating that Grb10 binding to Nedd4 is critical for ubiquitination of the receptor. These results suggest a role for the Grb10/Nedd4 complex in regulating ubiquitination and stability of the IGF-IR, and they suggest that Grb10 serves as an adapter to form a bridge between Nedd4 and the IGF-IR. This is the first demonstration of regulation of stability of a tyrosine kinase receptor by the Nedd4 (HECT) family of E3 ligases.


Roles of vasoconstrictor prostaglandins, COX-1 and -2, and AT1, AT2, and TP receptors in a rat model of early 2K,1C hypertension.
Angiotensin (ANG) II activating type 1 receptors (AT(1)Rs) enhances superoxide anion (O(2)*(-)) and arachidonate (AA) formation. AA is metabolized by cyclooxygenases (COXs) to PGH(2), which is metabolized by thromboxane (Tx)A(2) synthase to TxA(2) or oxidized to 8-isoprostane PGF(2alpha) (8-Iso) by O(2)*(-). PGH(2), TxA(2), and 8-Iso activate thromboxane-prostanoid receptors (TPRs). We investigated whether blood pressure in a rat model of early (3 wk) two-kidney, one-clip (2K,1C) Goldblatt hypertension is maintained by AT(1)Rs or AT(2)Rs, driving COX-1 or -2-dependent products that activate TPRs. Compared with sham-operated rats, 2K,1C Goldblatt rats had increased mean arterial pressure (MAP; 120 +/- 4 vs. 155 +/- 3 mmHg; P < 0.001), plasma renin activity (PRA; 22 +/- 7 vs. 48 +/- 5 ng x ml(-1) x h(-1); P < 0.01), plasma malondialdehyde (1.07 +/- 0.05 vs. 1.58 +/- 0.16 nmol/l; P < 0.01), and TxB(2) excretion (26 +/- 4 vs. 51 +/- 7 ng/24 h; P < 0.01). Acute graded intravenous doses of benazeprilat (angiotensin-converting enzyme inhibitor) reduced MAP at 20 min (-36 +/- 5 mmHg; P < 0.001) and excretion of TxA(2) metabolites. Indomethacin (nonselective COX antagonist) or SC-560 (COX-1 antagonist) reduced MAP at 20 min (-25 +/- 5 and -28 +/- 7 mmHg; P < 0.001), whereas valdecoxib (COX-2 antagonist) was ineffective (-9 +/- 5 mmHg; not significant). Losartan (AT(1)R antagonist) or SQ-29548 (TPR antagonist) reduced MAP at 150 min (-24 +/- 6 and -22 +/- 3 mmHg; P < 0.001), whereas PD-123319 (AT(2)R antagonist) was ineffective. Acute blockade of TPRs, COX-1, or COX-2 did not change PRA, but TxB(2) generation by the clipped kidney was reduced by blockade of COX-1 and increased by blockade of COX-2. 2K,1C hypertension in rats activates renin, O(2)*(-), and vasoconstrictor PGs. Hypertension is maintained by AT(1)Rs and by COX-1, but not COX-2, products that activate TPRs.


How to prepare cytological samples for molecular testing.
This review is focused on the challenges in standardising and optimising molecular testing workflow in cytopathology. Although cytological samples yield optimal quality DNA, whose minimal amounts in most cases suffice even for multigene mutational profiling, the success of molecular testing is strongly dependent on standardised preanalytical protocols for maximising DNA yield and quality. Sample cytopreparation influences, even more, the quality of RNA and consequently the potential success of reverse transcription-PCR. Here, the educational and technical involvement of the cytopathologist as a relevant component of a multidisciplinary team, in the issues related to test request, specimen collection, fixation, processing, staining, tumour fraction enrichment, DNA quality/quantity assessment and storage conditions is discussed. In addition, the specific sample requirements related to more recent technological developments are examined, underlining the modern role of the cytopathologist, whose continuous education is crucial to meet the opportunities of molecular medicine.


Repetitive Transcranial Magnetic Stimulation for Treatment-Resistant Depression: A Systematic Review and Meta-Analysis of Randomized Controlled Trials.
To date, several randomized controlled trials (RCTs) have shown the efficacy of repetitive transcranial magnetic stimulation (rTMS) in the treatment of major depression. This analysis examined the antidepressant efficacy of rTMS in patients with treatment-resistant unipolar depression. A literature search was performed for RCTs published from January 1, 1994, to November 20, 2014. The search was updated on March 1, 2015. Two independent reviewers evaluated the abstracts for inclusion, reviewed full texts of eligible studies, and abstracted data. Meta-analyses were conducted to obtain summary estimates. The primary outcome was changes in depression scores measured by the Hamilton Rating Scale for Depression (HRSD), and we considered, a priori, the mean difference of 3.5 points to be a clinically important treatment effect. Remission and response to the treatment were secondary outcomes, and we calculated number needed to treat on the basis of these outcomes. We examined the possibility of publication bias by constructing funnel plots and by Begg's and Egger's tests. A meta-regression was undertaken to examine the effect of specific rTMS technical parameters on the treatment effects. Twenty-three RCTs compared rTMS with sham, and six RCTs compared rTMS with electroconvulsive therapy (ECT). Trials of rTMS versus sham showed a statistically significant improvement in depression scores with rTMS (weighted mean difference [WMD] 2.31, 95% CI 1.19-3.43; P < .001). This improvement was smaller than the pre-specified clinically important treatment effect. There was a 10% absolute difference between rTMS and sham in the rates of remission or response. This translates to a number needed to treat of 10. Risk ratios for remission and response were 2.20 (95% CI 1.44-3.38, P = .001 and 1.72 [95% CI], 1.13-2.62, P = .01), respectively, favouring rTMS. No publication bias was detected. Trials of rTMS versus ECT showed a statistically and clinically significant difference between rTMS and ECT in favour of ECT (WMD 5.97, 95% CI 0.94-11.0, P = .02). Risk ratios for remission and response were 1.44 (95% CI 0.64-3.23, P = .38) and 1.72 (95% CI 0.95-3.11, P = .07), respectively, favouring ECT. Overall, the body of evidence favoured ECT for treatment of patients who are treatment-resistant. Repetitive transcranial magnetic stimulation had a small short-term effect for improving depression in comparison with sham, but follow-up studies did not show that the small effect will continue for longer periods.


An evolutionary algorithm for the discovery of porous organic cages.
The chemical and structural space of possible molecular materials is enormous, as they can, in principle, be built from any combination of organic building blocks. Here we have developed an evolutionary algorithm (EA) that can assist in the efficient exploration of chemical space for molecular materials, helping to guide synthesis to materials with promising applications. We demonstrate the utility of our EA to porous organic cages, predicting both promising targets and identifying the chemical features that emerge as important for a cage to be shape persistent or to adopt a particular cavity size. We identify that shape persistent cages require a low percentage of rotatable bonds in their precursors (<20%) and that the higher topicity building block in particular should use double bonds for rigidity. We can use the EA to explore what size ranges for precursors are required for achieving a given pore size in a cage and show that 16 Å pores, which are absent in the literature, should be synthetically achievable. Our EA implementation is adaptable and easily extendable, not only to target specific properties of porous organic cages, such as optimal encapsulants or molecular separation materials, but also to any easily calculable property of other molecular materials.


High resolution metabolite imaging in the hippocampus following neonatal exposure to the environmental toxin BMAA using ToF-SIMS.
The environmental neurotoxin β-N-methylamino-L-alanine (BMAA) is suggested to be linked with neurodegenerative disease. In a rat model, neonatal exposure to BMAA induced selective uptake in the hippocampus and caused cell loss, mineralization and astrogliosis as well as learning and memory impairments in adulthood. Moreover, neonatal exposure resulted in increased protein ubiquitination in the cornus ammonis 1 (CA1) region of the adult hippocampus indicating that BMAA may induce protein aggregation. Time-of-flight secondary ion mass spectrometry (ToF-SIMS) based imaging is a powerful technology for spatial profiling of small molecular weight compounds in biological tissues with high chemical specificity and high spatial resolution. The aim of this study was to characterize neurochemical changes in the hippocampus of six month-old rats treated neonatally (postnatal days 9-10) with BMAA. Multivariate data analysis of whole section ToF-SIMS scans was performed to delineate anatomical regions of interest based on their chemical distribution pattern. Further analysis of spectral data obtained from the outlined anatomical regions, including CA1 and dentate gyrus (DG) revealed BMAA-induced long-term changes. Increased levels of phospholipids and protein fragments in the histopathologically altered CA1 region as well as phosphate depletion in the DG were observed. Moreover, high resolution SIMS imaging revealed a specific localization of phosphatidylcholine lipids, protein signals and potassium in the histopathologically altered CA1. These findings demonstrate that ToF-SIMS based imaging is a powerful approach for probing biochemical changes in situ and might serve as promising technique for investigating neurotoxin-induced brain pathology.


Methods to improve efficacy of intravesical mitomycin C: results of a randomized phase III trial.
Intravesical chemotherapy (i.e., placement of the drug directly in the bladder) with mitomycin C is beneficial for patients with superficial bladder cancer who are at high risk of recurrence, but standard therapy is empirically based and patient response rates have been variable, in part because of inadequate drug delivery. We carried out a prospective, two-arm, randomized, multi-institutional phase III trial to test whether enhancing the drug's concentration in urine would improve its efficacy. Patients with histologically proven transitional cell carcinoma and at high risk for recurrence were eligible for the trial. Patients in the optimized-treatment arm (n = 119) received a 40-mg dose of mitomycin C, pharmacokinetic manipulations to increase drug concentration by decreasing urine volume, and urine alkalinization to stabilize the drug. Patients in the standard-treatment arm (n = 111) received a 20-mg dose without pharmacokinetic manipulations or urine alkalinization. Both treatments were given weekly for 6 weeks. Primary endpoints were recurrence and time to recurrence. Treatment outcome was examined by use of Kaplan-Meier analysis with log-rank tests. Statistical tests were two-sided. Patients in the two arms did not differ in demographics or history of intravesical therapy. Dysuria occurred more frequently in the optimized arm but did not lead to more frequent treatment termination. In an intent-to-treat analysis, patients in the optimized arm showed a longer median time to recurrence (29.1 months; 95% confidence interval [CI] = 14.0 to 44.2 months) and a greater recurrence-free fraction (41.0%; 95% CI = 30.9% to 51.1%) at 5 years than patients in the standard arm (11.8 months; 95% CI = 7.2 to 16.4 months) and 24.6% (95% CI = 14.9% to 34.3%) (P =.005, log-rank test for time to recurrence). Improvements were found in all risk groups defined by tumor stage, grade, focality, and recurrence. This study identified a pharmacologically optimized intravesical mitomycin C treatment with statistically significantly enhanced efficacy.


[Apoptosis: current status of the problem].
Accelerating progress in studying the nature of apoptosis (programmed cell death, PCD) has been observed in recent years. Apoptosis proved to be preprogrammed biological phenomenon with the key role in fundamental processes such as embryogenesis, morphogenesis, and cellular homeostasis. Apoptosis is induced by specific physiological signals as well as by various damaging influences. A complex system of metabolic and genetic control of apoptosis is realized by phylogenetically conserved set of genes with pro- and antiapoptotic activity. Genetic and metabolic disturbances of initiation and/or realization of PCD make substantial contribution to a number of pathologies such as cancer, AIDS, and degenerative processes. Active search for apoptosis-controlling remedies may open new horizons for fighting these widespread diseases.


College Students and SNAP: The New Face of Food Insecurity in the United States.
Over the last decade, multiple studies of food insecurity among college students have found rates from 20% to more than 50%, considerably higher than the 12% rate for the entire US population.Reasons for higher rates of food insecurity among college students include a growing population of low-income college students, high college costs and insufficient financial aid, more financial hardship among many low- and moderate-income families, a weak labor market for part-time workers, declining per capita college resources, and Supplemental Nutritional Assistance Program (SNAP) policies that specifically exclude many college students from participation.This essay reviews the causes and consequences of food insecurity on campus, explores reasons for the low SNAP participation rate, and describes how campuses have responded to food insecurity. It summarizes federal, state, and local changes in SNAP policies that can facilitate college student participation and retention and suggests strategies for more robust and effective university responses to food insecurity, including SNAP enrollment campaigns, a stronger role for campus food services, and a redefinition of the goals and purposes of campus food pantries.


The enzymic degradation of L-serine O-sulphate. Mechanism of the reaction.
1. During the enzyme-catalysed degradation of l-serine O-sulphate no exchange occurs between the hydrogen atom attached to the alpha-carbon atom of the substrate and the tritiated water of the incubation medium. 2. The participation of an intermediate carbanion has been demonstrated in the degradation by performing the reaction in the presence of tetranitromethane. 3. Photo-oxidation of the enzyme in the presence of Rose Bengal led to a rapid inactivation of enzyme with the concomitant loss of four histidine residues/molecule. 4. Rose Bengal was also a non-competitive inhibitor of the enzyme.


Measuring the quality of multi-disciplinary thoracic oncology care.
As a result of the rapid acceleration in new treatment advances in thoracic oncology, coordination and tailored selection of treatment modalities has become complex. This increasing complexity mandates multidisciplinary input and coordinated management. However, quality of care in the domain of multidisciplinary thoracic oncology has yet to be purposefully defined, and as a result the science of performance measurement has yet to be applied to this area. In this review we discuss the role of multi-disciplinary care in thoracic oncology, and outline its definitions as proposed in the literature. We review various metrics of quality in complex multidisciplinary thoracic oncology care, and future directions for quality and performance measurement.


The biosynthesis of deoxyribonucleic acid during the life-span of the mosquito.
Previous results from this laboratory [Kao et al, (1976) Biochem. J. 154, 471-480] demonstrated that a low-molecular-weight DNA species, "sDNA", present in high concentrations during rapid larval growth of the yellow-fever mosquito, Aedes aegypti, may be an intermediate in DNA replication. We have now determined the life-span profiles of synthetic rates and the steady-state contents of sDNA and total DNA by radiometric and biochemical procedures and by using mosquitoes grown and maintained under defined and controlled conditions, including axenic culture and synchronization of stages. 2. Our results showed exponential increases in DNA contents and synthetic rates which paralleled the changes in weight during larval growth. Of special importance was the finding that in rapidly-growing larvae and newly-emerged adult mosquitoes high contents and rates of synthesis of sDNA were correlated with increases in total DNA/organism, which suggests that sDNA content may be an index of DNA replication. In senescent mosquitos a twofold increase in the synthetic rate of bulk DNA ("pDNA") without an increase in sDNA may reflect repair synthesis. 3. The significance of this investigation is that it may be the first quantitative and systematic description of the synthetic rates and contents of both DNA-replication intermediates and total DNA in an axenic, eukaryotic organism rather than a specific cell type. Equally important is that these findings describe DNA changes during growth and also during aging through senescence in an age-characterized organism.


Catalases mediate tobacco resistance to virus infection through crosstalk between salicylic acid and auxin signaling pathways.
Catalases (CATs) play important roles in plant growth, development and defense responses. Previous studies have shown that CATs exhibit different or even opposite effects on plant immunity in different plant-pathogen interactions, but little is known about the mechanisms. In this study, Nicotiana tabacum plants with overexpression or knockout of CAT genes, tobacco mosaic virus (TMV) and cucumber mosaic virus (CMV) were employed to investigate the role of CAT in compatible plant-virus interactions. The results showed that there were dynamic changes in the effect of CAT on N. tabacum defense responses. Overexpression of catalase 1 (CAT1) and catalase 3 (CAT3) improved N. tabacum resistance in the early stage of virus infection but depressed it during the late stages of pathogenesis, especially in CAT3 overexpressing plants. The lower level of electrolyte leakage, lower contents of malonaldehyde and hydrogen peroxide (H2 O2 ), higher activities of antioxidant enzymes and improved functions of photosystem II corresponded to the milder symptoms and higher resistance of infected tobacco plants. In addition, the infection of TMV and CMV resulted in expression changes of CATs in tobacco plants, and pretreatment with H2 O2 facilitated TMV and CMV infection. Further experiments showed that the content of salicylic acid (SA) and the expression of genes related to SA signaling pathway were positively correlated with plant resistance, whereas auxin and its related signaling pathway were related to the viral susceptibility of plants. Taken together, our results demonstrated that CAT1 and CAT3 mediated tobacco resistance to virus infection through crosstalk between SA and auxin signaling pathways.


Droplet digital PCR technology promises new applications and research areas.
Digital Polymerase Chain Reaction (dPCR) is used to quantify nucleic acids and its applications are in the detection and precise quantification of low-level pathogens, rare genetic sequences, quantification of copy number variants, rare mutations and in relative gene expressions. Here the PCR is performed in large number of reaction chambers or partitions and the reaction is carried out in each partition individually. This separation allows a more reliable collection and sensitive measurement of nucleic acid. Results are calculated by counting amplified target sequence (positive droplets) and the number of partitions in which there is no amplification (negative droplets). The mean number of target sequences was calculated by Poisson Algorithm. Poisson correction compensates the presence of more than one copy of target gene in any droplets. The method provides information with accuracy and precision which is highly reproducible and less susceptible to inhibitors than qPCR. It has been demonstrated in studying variations in gene sequences, such as copy number variants and point mutations, distinguishing differences between expression of nearly identical alleles, assessment of clinically relevant genetic variations and it is routinely used for clonal amplification of samples for NGS methods. dPCR enables more reliable predictors of tumor status and patient prognosis by absolute quantitation using reference normalizations. Rare mitochondrial DNA deletions associated with a range of diseases and disorders as well as aging can be accurately detected with droplet digital PCR.


"Learning to learn" in the Extended Inverted Classroom: An evaluation of the effects of interactive teaching on knowledge and cognitive regulation in medical students.
The evidence regarding the effects of the Inverted Classroom on students' metacognitive skills is limited. This study evaluates these effects on student perceptions about knowledge and cognitive regulation during a surgical course. We measured student perceptions before and after a conventional Inverted Classroom in surgery using the Metacognitive Awareness Inventory. We evaluated the mean differences between the scores of the two measurements using Student's t-test (p < 0.05), and the size of the effect on knowledge and cognitive regulation using Cohen's d test (95%CI). A total of 158 students (pre-intervention) and 155 students (post-intervention) were included in the analysis. Significant differences were found (p < 0.05) in, as well as positive effects on, the students' abilities to know their own learning (declarative knowledge) (d = 0.24; 95%CI, 0.02-0.47), to use learning strategies (procedural knowledge) (d = 0.19; 95%CI, -0.02 to 0.41) and to understand when and why to use these strategies (conditional knowledge) (d = 0.20; 95%CI, -0.01 to 0.42), in addition to the abilities to plan (d = 0.31; 95%CI, 0.09-0.53) and to evaluate their learning (d = 0.31; 95%CI, 0.08-0.53). The Inverted Classroom has positive effects on metacognitive skills according to students' perceptions. Further studies are required that compare these effects with other methodologies for teaching in the classroom and the workplace.


The pleiotropic ts15 mutation of E. coli is an IS1 insertion in the rho structural gene.
Rho protein regulates transcription termination in E. coli. Some of the temperature-sensitive mutants defective in Rho protein, e.g., ts15, show remarkable pleiotropic phenotypes. The ts mutations map between the ilv and cya loci on the E. coli chromosome. We have cloned the gene that restores the wild-type phenotypes of these mutants. Genetic and biochemical characterizations have shown that the cloned DNA segment carries the structural gene for the Rho polypeptide. Analysis of the rhots15 mutation has revealed the presence of an IS1 insertion in the carboxy terminal segment of the rho cistron, thereby truncating the 52-kilodalton (kd) Rho polypeptide to a 50-kd size and also making it thermolabile. This provides an example of how an IS1 insertion mutation can cause a TS phenotype. We have also shown that the multiple phenotypes of the mutant cell, including the temperature sensitivity, are caused by a single mutation (rhots15::IS1) in the rho structural gene. How a rho structural gene mutation may cause such pleiotropy is discussed.


The association between mechanical temporal summation, state anxiety at baseline, and persistent low back pain: a 12-month prospective cohort study.
Persons with acute low back pain (LBP) have a good prognosis for regaining function, while pain often persists. Neurobiological and psychosocial factors are recognized to amplify pain responses, as reported for central sensitization. This study investigated the combination of mechanical temporal summation (TS) chosen to characterize central sensitization and state anxiety representing a psychological factor and their association with persistent pain. A longitudinal prospective cohort study including 176 participants aged between 18 and 65 with acute LBP was performed. The following independent variables were analyzed at baseline: The mechanical TS at the lower back, of whom the Wind-up ratio (WUR) was calculated, and the state anxiety level measured with the State and Trait Anxiety Inventory (STAI-S). The outcome pain intensity was assessed at baseline and 2,3,6 and 12 months after the onset of acute LBP with the Numeric Rating Scale 0-10 (NRS). Linear mixed models (LMM) were used to analyze the association of the independent variables with pain intensity over time. The mean baseline WUR was 1.3 (SD 0.6) for the right and 1.5 (SD 1.0) for the left side. STAI-S revealed a mean score of 43.1 (SD 5.2). Pain intensity was, on average, 5.4 points (SD 1.6) on the NRS and decreased over one year to a mean of 2.2 (SD 2.4). After one year, 56% of the participants still experienced pain. The LMM revealed a considerable variation, as seen in large confidence intervals. Therefore, associations of the independent variables (WUR and STAI-S) with the course of the outcome pain intensity over one year were not established. This investigation did not reveal an association of mechanical TS and state anxiety at baseline with pain intensity during the one-year measurement period. Pain persistence, mediated by central sensitization, is a complex mechanism that single mechanical TS and state anxiety cannot capture.


Arrestin/clathrin interaction. Localization of the arrestin binding locus to the clathrin terminal domain.
Previously we demonstrated that nonvisual arrestins exhibit a high affinity interaction with clathrin, consistent with an adaptor function in the internalization of G protein-coupled receptors (Goodman, O. B., Jr., Krupnick, J. G., Santini, F., Gurevich, V. V., Penn, R. B., Gagnon, A. W., Keen, J. H., and Benovic, J. L. (1996) Nature 383, 447-450). In this report we show that a short sequence of highly conserved residues within the globular clathrin terminal domain is responsible for arrestin binding. Limited proteolysis of clathrin cages results in the release of terminal domains and concomitant abrogation of arrestin binding. The nonvisual arrestins, beta-arrestin and arrestin3, but not visual arrestin, bind specifically to a glutathione S-transferase-clathrin terminal domain fusion protein. Deletion analysis and alanine scanning mutagenesis localize the binding site to residues 89-100 of the clathrin heavy chain and indicate that residues 1-100 can function as an independent arrestin binding domain. Site-directed mutagenesis identifies an invariant glutamine (Glu-89) and two highly conserved lysines (Lys-96 and Lys-98) as residues critical for arrestin binding, complementing hydrophobic and acidic residues in arrestin3 which have been implicated in clathrin binding (Krupnick, J. G., Goodman, O. B., Jr., Keen, J. H., and Benovic, J. L. (1997) J. Biol. Chem. 272, 15011-15016). Despite exhibiting high affinity clathrin binding, arrestins do not induce coat assembly. The terminal domain is oriented toward the plasma membrane in coated pits, and its binding of both arrestins and AP-2 suggests that this domain is the anchor responsible for adaptor-receptor recruitment to the coated pit.


A randomized controlled trial comparing zileuton with theophylline in moderate asthma. The Zileuton Study Group.
Zileuton, a leukotriene pathway inhibitor, was compared with slowly absorbed theophylline in a randomized, double-blind study of patients with chronic asthma. The primary efficacy measure was improvement in forced expiratory volume in 1 second (FEV1). Eligibility criteria included FEV1 of 40% to 80% of predicted, documented reversibility of airway disease, and age 18 to 60 years. Initially, the theophylline dosage was titrated to achieve trough concentrations of 8 to 15 micrograms/mL. After washout and 1-week placebo lead-in, patients were randomly assigned to 13 weeks of the appropriate theophylline dose or zileuton, 400 or 600 mg 4 times daily. The FEV1 was measured before the morning dose at 2-week intervals and serially after the dose on days 36 and 92. Patients kept daily diaries of asthma symptoms, beta-agonist usage, and peak expiratory flow rate; on days 36 and 92, they completed quality-of-life questionnaires. Of 471 eligible patients at 38 centers, 377 were randomly assigned to the study; 313 completed the study. On first-dose administration, all groups showed 11% to 13% improvement in FEV1 within 30 minutes. Patients who received zileuton, 400 mg, had significantly greater improvement at several points than did theophylline-treated patients. The range of long-term maximum improvement in FEV1 in the groups was 30% to 34% (P = .40 for zileuton 600 mg; P = .90 for zileuton 400 mg vs theophylline). Initially, the theophylline group improved significantly more in symptom scores, beta-agonist usage, and peak expiratory flow rate, but at maximal effect there was no significant difference. All groups showed significant improvement in quality of life. No overall differences were observed between the zileuton dosage groups. Adverse events were comparable in all groups. Zileuton appears as effective and safe as theophylline in patients with chronic asthma.


cCXCR1 is a receptor for cIL-8 (9E3/cCAF) and its N- and C-terminal peptides and is also activated by hIL-8 (CXCL8).
Chemokines are chemotactic cytokines that play important roles in immune responses and wound healing, as well as in pathological conditions such as chronic inflammation and tumorigenesis. The chemokines and their receptors are highly conserved and maintain similar functions in different species. One noteworthy exception is the chemokine interleukin (IL)8/CXC ligand 8 and its specific receptor CXCR1, which are found in humans but are not found in the traditional model organisms, mice and rats. As a consequence, we are using model organisms other than mice to study the functions of IL-8 and CXCR1, as well as the mechanisms involved in receptor activation by IL-8. Toward this goal, we have isolated and characterized a new receptor that is highly homologous to human (h)CXCR1, which we named chicken (c)CXCR1. To determine whether this receptor is activated by cIL-8 and its N- and C-terminal peptides and whether it responds to hIL-8, we expressed cCXCR1 in NIH3T3 cells, which naturally lack this receptor, and used single-cell Ca(2)(+) imaging to detect increases in intracellular Ca(2)(+) and immunoblot analysis to detect extracellular signal-regulated kinase 1/2 phosphorylation. We show that cIL-8, its N and C peptides, and hIL-8 activate cCXCR1. We further show that cIL-8 and hIL-8 stimulate chemotaxis of chicken embryonic fibroblasts, cells that express cCXCR1, and that this effect is specific for each chemokine and this receptor. These results strongly suggest that cCXCR1 is the ortholog for hCXCR1 and that chickens can be used as an effective model system to study the functions of IL-8, its terminal peptides, and its specific receptor CXCR1.


Double fertilization in ephedra, a nonflowering seed plant: its bearing on the origin of angiosperms.
Double fertilization and the associated formation of endosperm have long been considered unique and defining characters (autapomorphies) of the angiosperms. During normal fertilization in Ephedra nevadensis, a nonflowering seed plant, fusion of a second sperm nucleus with the ventral canal nucleus occurs regularly within the egg cytoplasm. The occurrence of double fertilization in Ephedra assumes added significance in light of its critical phylogenetic position as a basal member of the most closely related extant group of seed plants (Gnetales) to angiosperms. Thus, double fertilization in angiosperms and Ephedra may represent an evolutionary homology.


The Association between Oxytocin and Lower Limb Osteoarthritis: A Prospective Cohort Study.
Oxytocin (OT), a neuropeptide best known for its role in emotional and social behaviors, has been linked to osteoarthritis (OA). This study aimed to investigate the serum OT level in hip and/or knee OA patients and to study its association with disease progression. Patients from the KHOALA cohort with symptomatic hip and/or knee OA (Kellgren and Lawrence (KL) scores of 2 and 3) and follow-up at 5 years were included in this analysis. The primary endpoint was structural radiological progression, which was defined as an increase of at least one KL point at 5 years. Logistic regression models were used to estimate the associations between OT levels and KL progression while controlling for gender, age, BMI, diabetes and leptin levels. Data from 174 hip OA patients and 332 knee OA patients were analyzed independently. No differences in OT levels were found between the 'progressors' and 'non-progressors' groups among the hip OA patients and knee OA patients, respectively. No statistically significant associations were found between the OT levels at baseline and KL progression at 5 years, the KL score at baseline or the clinical outcomes. Higher structural damage at baseline and severe structural progression of hip and knee osteoarthritis did not appear to be associated with a low serum OT level at baseline.


Narrative review of pembrolizumab for the treatment of esophageal cancer: evidence and outlook.
Based on the current evidence, review the efficacy and safety profile of pembrolizumab, along with its shortcomings, in an effort to define future research directions. The survival outcome of esophageal cancer (EC) is poor, especially in patients with advanced stage. Palliative surgery, chemotherapy, radiotherapy and chemoradiotherapy have limited efficacy in prolonging the survival time. Currently, immunotherapies, including adoptive cell therapy-based, antibody-based, and vaccine-based therapies, are attracting considerable attention. The mechanism of immunotherapy lies in the modification of immune response and prevention of immune escape. Immunomodulatory agents can block the programmed cell death protein 1/programmed death-ligand 1 (PD-1/PD-L1) pathway, thereby allowing lymphocytes to attack tumor cells. This class of drugs has the potential to treat a variety of tumors and may substantially improve overall survival (OS) in some patients. Multiple clinical trials have shown that pembrolizumab has good efficacy and safety, enhances the EC treatment paradigm, and has even become the first-line treatment of choice for patients with PD-L1-positive recurrent or metastatic EC. We reviewed the results of clinical trials of pembrolizumab for EC and gastroesophageal cancer presented at Embase, PubMed, the American Society of Clinical Oncology (ASCO) annual meetings, and the Cochrane Central Register of Controlled Trials. Pembrolizumab has good efficacy and tolerability profiles, and has emerged as a second-line option for the treatment of PD-L1-positive locally advanced or metastatic ESCC. Pembrolizumab has many promising applications, and further investigations into its mechanisms should be conducted.


Molecular characterization and genetic susceptibility of sapovirus in children with diarrhea in Burkina Faso.
Sapoviruses (SaVs) are a common cause of gastroenteritis in children. In sub-Saharan Africa, there is a scarcity of information regarding SaV as an etiological agent of diarrhea. Here, we investigated the prevalence, molecular characterization and clinico-epidemiological features of SaV infections in children less than 5years of age with diarrhea in Burkina Faso. We further investigated the role of type 1 histo blood group antigens as susceptibility factors. In total, 309 fecal and 208 saliva samples from diarrheal children in Ouagadougou, Burkina Faso, were collected between May 2009 and March 2010. SaV was detected using real-time PCR, and genogrouped/genotyped by PCR or sequencing. Saliva samples were ABO, Lewis and secretor phenotyped using in house ELISA assays. We found a high prevalence (18%) and large genetic diversity with all 4 human genogroups, and 9 genotypes/genoclusters circulating during the study period. The SaV infections were generally associated with milder symptoms, and neither ABH, Lewis or secretor phenotypes affected susceptibility to SaV infections.


A One-Year Introductory Biology Majors' Lab Sequence Incorporating Vision & Change.
The introduction of Vision and Change by AAAS and the recommendation that biology departments amend their curricula to focus on key concepts and skills necessary for graduates have led to a re-envisioning of introductory curricula across the nation. Many of the "standard" biology text books have realigned their focus with Vision and Change, while new texts have emerged that completely revise how we teach introductory biology majors. One such textbook is Integrating Concepts in Biology (ICB), by Campbell, Heyer, and Paradise. Many departments, including ours, have adopted this text as a novel way to teach biology majors, focusing on active learning, the scientific method, and specifically, understanding data. However, with all of these revisions to biology textbooks, there have been no revisions or insights into corresponding labs for a typical 1-year introductory course sequence. Here, we provide a description of our 1-year lab sequence, emphasizing the scientific method and novel research, with a focus on the five "Big Ideas" presented in ICB. By removing the "cookbook" labs typical of most introductory laboratory courses, we found that this system better emphasized the focus of Vision and Change and, concomitantly, student appeared to enjoy the lab sequence and see the relevance to class material better, compared to previous years. We believe that this lab organization is a simple design that is not resource-intensive and can be utilized at schools of any size or budget.


[Half century of botany publishing in Revista de Biologia Tropical].
Over its first half century the Revista de Biología Tropical published many papers and supplements dealing with the botany. However, the Revista is not a primary botanical journal. A wide variety of topics and geographic sources have been included, taking into consideration species from the Neotropics, but also from India and Nigeria. A complete index of botanical papers is presented.


Skin cancer incidence is highly associated with ultraviolet-B radiation history.
Recently, the increased amount of ultraviolet-B (UV-B) exposure due to ozone depletion has been found to be associated with increased incidence of skin cancer across the world. The quantification of individual, regional, and historical UV exposure directly affects establishment of the association between skin cancer and UV exposure, but accurate assessment and measurement have been challenging for decades. As a sequence, cumulative studies using different metrics reported conflicting results on whether UV radiation, including sunburns, early childhood sun exposure, and chronic exposure, increases melanoma risk. This paper aims to establish the relationship between UV-B and melanoma incidence across the continental U.S. using an ecological approach that incorporate more accurate UV-B exposure measured by the National Aeronautical and Space Administration Nimbus-7 total ozone mapping spectrometer, and the United State Department of Agriculture ground-based network. Using statistical linear mixed models, we found strong positive associations between the skin cancer and the past UV exposure or the past cumulative 3-year UV exposure 3 or 4 years ago. UV has regional distributions and its regional effects on the skin cancer incidence are still significant after adjusting the effect of UV exposure. Research findings yield deepened understanding of spatiotemporal distribution of melanoma incidence rates and a greater appreciation for the complexity and heterogeneity of melanoma risk factors especially the UV-B exposure at different temporal and spatial scales.


Frontal lobe volume in patients with Huntington's disease.
Neuropathologic and neuroimaging studies have suggested that frontal lobes are affected in Huntington's disease (HD), and that atrophy in this region may be associated with some of the cognitive impairment and clinical decline observed in patients with HD. We measured gray and white matter volumes within the frontal lobes on MRI for 20 patients with HD (10 mildly affected and 10 moderately affected) and 20 age- and sex-matched control subjects. We also correlated frontal lobe measurements with measures of symptom severity and cognitive function. Patients who were mildly affected had frontal lobe volumes (both gray and white matter) essentially identical to those of control subjects, despite clearly abnormal basal ganglia. Patients who were moderately affected demonstrated significant reductions in total frontal lobe volume (17%) and frontal white matter volume (28%). Frontal lobe white matter volume reductions, but not total frontal lobe volume reductions, were disproportionately greater than overall brain volume reductions (17%). Frontal lobe volume correlated with symptom severity and general cognitive function, but these correlations did not remain significant after taking into account total brain volume. We conclude that cognitive impairment and symptom severity are associated with frontal lobe atrophy, but this association is not specific to the frontal lobes. Frontal lobe atrophy (like total brain atrophy) occurs in later stages of increasing HD symptom severity and this atrophy primarily involves white matter.


Hospital influences on early infant-feeding practices.
Newborn nursery nursing staff members were surveyed to determine their attitudes and teaching practices regarding breast- and bottle-feeding. Concurrently, mothers using this nursery responded to a structured interview concerning their infant-feeding practices at 14 to 21 days postpartum and possible hospital influences on these practices. The nursing staff strongly advocated breast-feeding and did not favor specific bottle-feeding practices or products. Nursing staff counseling was generally interpreted by mothers as supporting breast-feeding, but this did not deter a large proportion of mothers who stated an initial preference for breast-feeding from introducing formula as a supplementary or exclusive form of infant feeding during the short study period. Almost all mothers doing any amount of bottle-feeding at the time of their interview were using the same formula brand and a ready-to-feed preparation used during their hospital stay. Other influences on mother's infant-feeding patterns are discussed. It is concluded that the hospital staff and routines exerted a stronger influence on mothers' infant-feeding practices by nonverbal teaching (the hospital "modeling" of infant formula products) than by verbal teaching (counseling supporting breast-feeding). Future studies might explore new ways of supporting mothers who desire to breast-feed by designing innovative hospital routines to model breast-feeding rather than feeding by infant formula.


Suramin prevents binding of interleukin 2 to its cell surface receptor: a possible mechanism for immunosuppression.
Suramin, a polysulfonic naphthalene antihelminthic drug, inhibits proliferation of a variety of T-cell lines in vitro and induces immunosuppression in some human patients and thymic atrophy and splenic depletion in mice. Recent clinical trials indicate that suramin has activity against human tumors, indicating that it will be necessary to understand the mechanism by which suramin induces immunosuppression. The T-cell growth factor, interleukin 2 (IL2), is the major growth factor involved in regulating lymphoid differentiation and proliferation and thus regulates, to a major degree, the magnitude and duration of the immune response. We demonstrate herein that suramin induces a concentration-dependent decrease in binding of 125I-labeled IL2 to its receptor complex on human and murine T-lymphocytes. Binding of 125I-labeled IL2 to both Mr 75,000 and 55,000 IL2 binding molecules was inhibited by suramin. Similar concentrations of suramin were required to inhibit binding of 125I-labeled IL2, IL2-induced tyrosine phosphorylation, and IL2-induced proliferation, suggesting that these processes may be linked. With murine cells, suramin-induced growth inhibition could be overcome completely by increasing the concentration of IL2, suggesting that suramin inhibited growth by competing for the IL2 receptor. With human cells, growth inhibition by suramin could only be partially overcome by increasing the concentration of IL2, suggesting that an additional growth-inhibiting mechanism is present. The ability of suramin to prevent binding of IL2 to its receptor was used to confirm that prolonged interaction of IL2 with its receptor is required to induce cell proliferation. Since IL2 plays a role in lymphocyte proliferation and differentiation, the ability of suramin to inhibit binding of IL2 to its receptor may explain, in part, the in vivo immunosuppressive activities of suramin.


The effect of exercise during haemodialysis on solute removal.
Urea rebound results as urea re-equilibrates between intracellular and intravascular compartments post haemodialysis. The mechanism of the rebound is thought to be due to either a reduced diffusion rate or blood flow. It is hypothesized that low blood flow in the skeletal muscles might be responsible. We tested this by studying the effect of exercise during dialysis on the removal of urea, creatinine and potassium. Eleven patients (aged 32-78 years) on haemodialysis (4-58 months) were studied on paired dialysis sessions; one with exercise and the other as a control. Patients pedalled on a cycle for 5-20 min at submaximal workload followed by 10 min rest to achieve a total of 60 min exercise. Plasma concentrations of urea, creatinine and potassium were measured pre-, post- and 30-min post dialysis. The post-dialysis rebound (% rebound) and reduction ratios (RR) of the solutes and equilibrated (two-pool) urea Kt/V were calculated for comparison. The rebound of all three solutes was reduced significantly following exercise. The rebound of urea decreased from 12.4 to 10.9% (median, P<0.01 Wilcoxon signed rank test), creatinine from 21.2 to 17.2% (P<0.001) and potassium from 62 to 44% (P<0.05). Kt/V and RR increased significantly as a result: Kt/V urea from 1.00 to 1.15 (P=0.001), RR urea from 0.63 to 0.68 (P<0.001); Kt/V creatinine from 0.71 to 0.84 (P<0.01); and RR creatinine from 0.51 to 0.57 (P<0.05). Exercise increased the efficiency of dialysis by reducing the rebound of solutes due to increased perfusion of the skeletal muscles.


Hair Bundle Stimulation Mode Modifies Manifestations of Mechanotransduction Adaptation.
Sound detection in auditory sensory hair cells depends on the deflection of the stereocilia hair bundle which opens mechano-electric transduction (MET) channels. Adaptation is hypothesized to be a critical property of MET that contributes to the auditory system's wide dynamic range and sharp frequency selectivity. Our recent work using a stiff probe to displace hair bundles showed that the fastest adaptation mechanism (fast adaptation) does not require calcium entry. Using fluid-jet stimuli, others obtained data showing only a calcium-dependent fast adaptation response. Because cochlear stereocilia do not move coherently and the hair cell response depends critically on the magnitude and time course of the hair bundle deflection, we developed a high-speed imaging technique to quantify this deflection in rat cochlear hair cells. The fluid jet delivers a force stimulus, and force steps lead to a complex time course of hair bundle displacement (mechanical creep), which affects the hair cell's macroscopic MET current response by masking the time course of the fast adaptation response. Modifying the fluid-jet stimulus to generate a hair bundle displacement step produced rapidly adapting currents that did not depend on membrane potential, confirming that fast adaptation does not depend on calcium entry. MET current responses differ with stimulus modality and will shape receptor potentials of different hair cell types based on their in vivo stimulus mode. These transformations will directly affect how stimuli are encoded.SIGNIFICANCE STATEMENT Mechanotransduction by sensory hair cells represents a key first step for the sound sensing ability in vertebrates. The sharp frequency tuning and wide dynamic range of sound sensation are hypothesized to require a mechanotransduction adaptation mechanism. Recent work indicated that the apparent calcium dependence of the fastest adaptation differs with the method of cochlear hair cell stimulation. Here, we reconcile existing data and show that calcium entry does not drive the fastest adaptation process, independent of the stimulation method. With force stimulation of the hair bundle, adaptation manifests differently than with displacement stimulation, indicating that the stimulation mode of the hair bundle will affect the hair cell receptor current and stimulus coding.


Effect of recombinant human granulocyte/macrophage colony-stimulating factor on neutrophil superoxide production.
Recombinant human granulocyte/macrophage colony-stimulating factor (GM-CSF) induced significant superoxide production in human neutrophils within 30 minutes after addition of stimulus and the response was complete within 2 hr. Other agents known to prime neutrophils, including LPS and tumor necrosis factor-alpha, lacked activity under the experimental conditions employed. Using a panel of pharmacologic inhibitors, we sought to compare GM-CSF-induced neutrophil superoxide to that produced by cells exposed to N-formyl methionyl-leucyl-phenylalanine (fMet-Leu-Phe) and phorbol 12-myristate 13-acetate (PMA). Each stimulant displayed a different profile. Rolipram, a peak IV phosphodiesterase inhibitor, specifically inhibited neutrophil activation by GM-CSF and fMet-Leu-Phe, while superoxide production stimulated by PMA was unaffected. Staurosporine, a protein kinase C (PK-C) inhibitor, suppressed superoxide production induced by all three neutrophil stimulants. Cytochalasin B totally inhibited superoxide induced by GM-CSF under conditions that promote the fMet-Leu-Phe-induced response. Cytochalasin B did not markedly affect PMA-induced superoxide. The results are consistent with the hypothesis that intact PK-C activity is essential for neutrophil superoxide production, but that differences exist in the initial pathways induced by these neutrophil activators. Superoxide secretion from GM-CSF-treated neutrophils appears to be a direct, delayed response that requires assembly of microfilaments during exposure to the cytokine.


Efficient Cu2O Photocathodes for Aqueous Photoelectrochemical CO2 Reduction to Formate and Syngas.
Photoelectrochemical carbon dioxide reduction (PEC-CO2R) represents a promising approach for producing renewable fuels and chemicals using solar energy. However, attaining even modest solar-to-fuel (STF) conversion efficiency often necessitates the use of costly semiconductors and noble-metal catalysts. Herein, we present a Cu2O/Ga2O3/TiO2 photocathode modified with Sn/SnOx catalysts through a simple photoelectrodeposition method. It achieves a remarkable half-cell STF efficiency of ∼0.31% for the CO2R in aqueous KHCO3 electrolyte, under AM 1.5 G illumination. The system enables efficient production of syngas (FE: ∼62%, CO/H2 ≈ 1:2) and formate (FE: ∼38%) with a consistent selectivity over a wide potential range, from +0.34 to -0.16 V vs the reversible hydrogen electrode. We ascribe the observed performance to the favorable optoelectronic characteristics of our Cu2O heterostructure and the efficient Sn/SnOx catalysts incorporated in the PEC-CO2R reactions. Through comprehensive experimental investigations, we elucidate the indispensable role of Cu2O buried p-n junctions in generating a high photovoltage (∼1 V) and enabling efficient bulk charge separation (up to ∼70% efficiency). Meanwhile, we discover that the deposited Sn/SnOx catalysts have critical dual effects on the overall performance of the PEC devices, serving as active CO2R catalysts as well as the semiconductor front contact. It could facilitate interfacial electron transfer between the catalysts and the semiconductor device for CO2R by establishing a barrier-free ohmic contact.


[Healthcare-associated urinary tract infections in patients with a urinary catheter: Risk factors, microbiological characteristics and patterns of antibiotic resistance].
Catheter-associated urinary tract infections (CAUTIs) are the most common nosocomial acquired infections, with high resistance rates. CAUTIs are a potentially severe complication in hospitalized patients and imply higher costs. Our aim was to analyze the characteristics of CAUTIs in our Urology department. Between November 2011 and October 2013, a prospective observational study was carried out analyzing the incidence of healthcare-associated urinary tract infections in patients admitted to the urology ward with an indwelling urinary catheter. Furthermore, we evaluated associated risks factors and comorbidities such as urinary catheter at the time of admission or urological surgery during the hospitalization. We also presented our results regarding the microbiological characteristics and patterns of resistance to antibiotics in patients with CAUTI admitted in our service. The incidence of CAUTI was 8.2% (189/2283 patients who had urinary catheter). Mean age was 67.4±14.26 years, 90.2% underwent a surgical procedure (p<0.001), 14.8% had a urinary stone (p=0.058) and 46% had a urinary catheter before admission (p<0.001). The most commonly isolated pathogens were Escherichia coli (22.6%), Enterococcus (21.9%) and Pseudomonas aeruginosa (13.9%). E.coli showed resistance rates of 41.9% for quinolones, 33.3% of them produced extended spectrum Β-lactamase (ESBL). P.aeruginosa showed resistance rates of 42.1% for quinolones and 21.1% for carbapenems. Healthcare-associated CAUTI in patients hospitalized in a urology ward are related to risks factors such as having a urinary catheter before admission and undergoing a surgical procedure. Moreover, CAUTIs have higher incidence of pathogens with antibiotic resistances and non-common pathogens such as Enterococcus spp.


Referral pathways between the mental health services and Jigsaw.
To examine similarities and differences in the demographic and clinical profiles of young people (15-25 years of age) referred between the mental health services (MHS) and Jigsaw Galway. A retrospective chart review was conducted of clinical files of individuals attending secondary MHS who had been referred to or from Jigsaw Galway over a 5-year period. Differences in demographic and clinical data between individuals referred to or from Jigsaw Galway were compared. A recent act of self-harm was more prevalent in individuals referred from Jigsaw to the adult MHS (p=0.02). No other demographic or clinical differences were detected between individuals attending Jigsaw Galway and the MHS. Education sessions for clinical staff working in primary care, Jigsaw Galway and the MHS are suggested to support clinicians in choosing the best referral pathway, which may more optimally address young people's mental health difficulties.


Neuropathic pain may be common in chronic lower limb tendinopathy: a prospective cohort study.
To identify the prevalence of neuropathic pain, through the use of the painDETECT questionnaire, in a cohort of patients with chronic lower limb tendinopathy conditions. Patients with chronic lower limb tendinopathy conditions treated within a Sport and Exercise Medicine hospital clinic were identified from clinical records. At the time of the clinical consultation, pain and painDETECT scores were recorded. In total, 282 suitable patients with chronic lower limb tendinopathy conditions were identified who had completed a painDETECT questionnaire. There was a median age of 51.9 years, 35% of patients were male and a median duration of symptoms of 24.0 months. There was a median score of 7.0/10 for self-reported 'average' pain and 8.0/10 for self-reported 'worst' pain. There was a median painDETECT score of 14.0, 28% of respondents scored 19 or higher with painDETECT (neuropathic component to pain may be likely), 29% scored 13-18 (equivocal result) and 43% of respondents scored 12 or less (neuropathic pain component was unlikely). This study suggests that neuropathic pain as identified by the painDETECT questionnaire may be common in patients with chronic lower limb tendinopathy conditions. It is unclear if patients with tendinopathy who have neuropathic pain may have poorer outcomes from initial treatments, contributing to the high proportion seen in secondary care. These are results from a single hospital clinic, and comparison with a control group is currently lacking. However, on the results to date, neuropathic pain should be considered in management strategies in patients with chronic tendinopathy.


Oral anticoagulation: a critique of recent advances and controversies.
There have recently been significant advances in the field of oral anticoagulation, but these have also led to many controversies. Warfarin is still the commonest drug used for clotting disorders but its use is complicated owing to wide inter-individual variability in dose requirement and its narrow therapeutic index. Warfarin dose requirement can be influenced by both genetic and environmental factors. Two recent randomized controlled trials (RCTs) came to different conclusion regarding the utility of genotype-guided dosing; we critically explore the reasons for the differences. The new generation of oral anticoagulants have been demonstrated to be as efficacious as warfarin, but further work is needed to evaluate their safety in real clinical settings.


Role of p16 testing in cervical cancer screening among HIV-infected women.
p16 immunohistochemistry is used to evaluate for HPV-associated cervical intraepithelial neoplasia. The diagnostic performance of p16 in HIV infection is unclear. Between June-December 2009, HIV-infected women underwent Papanicolaou (Pap) smear, human papillomavirus (HPV) testing, visual inspection with acetic acid (VIA), and colposcopy-directed biopsy as the disease gold standard at a HIV clinic in Kenya. Pap smears were evaluated for p16 expression. Sensitivity, specificity, positive predictive value (PPV), and area under the receiver operating characteristic curve (AUC) of p16 to detect CIN2/3 on histology and the impact of immunosuppression and ART was assessed. Of 331 cervical samples with p16 expression, p16 sensitivity and specificity to detect CIN2/3 was 54.1% and 72.4% respectively, which was lower than Pap and HPV in sensitivity, but higher in specificity than Pap, HPV, and VIA. Combining tests and p16 reduced sensitivity and increased specificity of Pap from 90.5% to 48.7% and 51.4% to 81.7%; of VIA from 59.5% to 37.8% and 67.6% to 89.9%; and of HPV from 82.4% to 50.0% and 55.3% to 84.8%. Combination p16 increased the PPV of Pap from 34.9% to 43.4%; of HPV from 34.7% to 48.7%; and VIA from 34.9% to 51.9%. Adjunctive p16 did not change AUC (P>0.05). P16 performance was not altered by immunosuppression or ART use. Combining p16 with HPV and VIA reduced the variation in HPV and VIA performance associated with CD4 and ART. As an adjunctive test in HIV-infected women, p16 immunohistochemistry increased specificity and PPV of HPV and VIA for CIN2/3, and was not altered in performance by immunosuppression, ART, or age.


Regulation of murine type 1 plasminogen activator inhibitor gene expression in vivo. Tissue specificity and induction by lipopolysaccharide, tumor necrosis factor-alpha, and transforming growth factor-beta.
The regulation of type 1 plasminogen activator inhibitor (PAI-1) gene expression was studied in vivo employing a murine model system. Nuclease protection analysis revealed relatively high concentrations of PAI-1 mRNA in the aorta, adipose tissue, heart, and lungs of untreated CB6 (BalbC X C57B16) mice. Treatment of CB6 mice with LPS, TNF-alpha, or transforming growth factor-beta (TGF-beta) increased the steady-state levels of PAI-1 mRNA within 3 h in all tissues examined. However, the greatest responses to TGF-beta were observed in adipose tissue and the kidney, while LPS and TNF-alpha strongly stimulated PAI-1 gene expression in the liver, kidney, lung, and adrenals. In C3H/HeJ mice, which exhibit defective TNF-alpha release in response to LPS, the response of the PAI-1 gene to LPS was severely attenuated. However, injection of these mice with TNF-alpha increased PAI-1 mRNA in a tissue-specific pattern strikingly similar to that observed in LPS-treated CB6 mice. These results demonstrate that the PAI-1 gene is regulated in a complex and tissue-specific manner in vivo, and suggest a role for TNF-alpha in the response of the PAI-1 gene to sepsis.


Implementation of an animated medication information tool in community pharmacies, with a special focus on patients with limited health literacy.
The animated medication information tool 'Watchyourmeds' provides information in an accessible manner through animated videos and therefore appears to be especially suitable for people with limited health literacy. This study aimed to assess the implementation of this animated medication information tool in Dutch community pharmacies, with a special focus on patients with limited health literacy. A cross-sectional survey based on the Reach, Effectiveness, Adoption, Implementation and Maintenance (RE-AIM) framework was sent to approximately 75% of the ±1900 community pharmacies in the Netherlands through email newsletters of pharmacy networks. 140 pharmacists (⁓10%) completed the survey and 125 of them (89%) indicated that they offered the animated medication information tool to their patients. 108 pharmacists indicated that the tool was offered to all patients, not only to patients with limited health literacy. The distribution method was primarily passive (patients were given a leaflet and were not explicitly pointed to or informed about the tool). Two frequently cited motivations for offering the tool were that it complemented other sources of information and that the health insurer provided a financial incentive. The main reasons patients refused to use the tool were that they had no access to or no affinity for the required technology. This study demonstrated that the tool is used in community pharmacies and that it is offered to all patients, regardless of their presumed health literacy level. A more active method of offering the tool may be warranted to better reach patients with limited health literacy.


Production and characterisation of monoclonal antibodies for the quantification of potentially allergenic xylanase from Aspergillus niger.
Xylanase from Aspergillus niger (ANX) is widely used in bakeries as a processing aid since it stabilises and improves dough quality. An association between allergic symptoms among bakery workers and sensitisation to ANX has been reported, indicating that this enzyme is an occupational allergen. The presence of ANX in dough improvers and semi-finished goods is often hidden due to incomplete and unclear labelling. The quantification of microbial enzymes in these products is necessary and the determination of the actual concentration of ANX in workplaces is therefore essential to assess the occupational risk. To this purpose we have developed and characterised monoclonal antibodies to ANX. The monoclonal antibodies do not show any cross-reaction with other commonly used microbial enzymes, and they allow the detection of ANX in complex mixtures by ELISA inhibition assays down to the concentration limit of approximately 10 µg kg(-1). These mAbs are a valuable tool to detect and quantify ANX and to investigate its allergenic potential in the workplace.


Hydroxylamine as an oxygen nucleophile: substitution of sulfonamide by a hydroxyl group in benzothiazole-2-sulfonamides.
Benzothiazole-2-sulfonamides react with an excess of hydroxylamine in aqueous solutions to form 2-hydroxybenzothiazole, sulfur dioxide, and the corresponding amine. Mechanistic studies that employ a combination of structure-reactivity relationships, oxygen labeling experiments, and (in)direct detection of intermediates and products reveal that the reaction proceeds via oxygen attack, and that oxygen incorporated in the 2-hydroxybenzothiazole product derives from hydroxylamine. The reaction, which is performed under mild conditions, can be used as a deprotection method for cleavage of benzothiazole-2-sulfonyl-protected amino acids.


Hypomagnesemia and its relation with chronic low-grade inflammation in obesity.
The accumulation of visceral fat in obesity is associated with excessive production of proinflammatory adipokines, which contributes to low-grade chronic inflammation state. Moreover, the literature has shown that mineral deficiency, in particular of magnesium, has important role in the pathogenesis of this metabolic disorder with relevant clinical repercussions. To bring updated information about the participation of hypomagnesemia in the manifestation of low-grade chronic inflammation in obese individuals. Articles published in PubMed, SciELO, LILACS and ScienceDirect, using the following keywords: "obesity," "magnesium" and "low grade inflammation." Scientific evidence suggests that magnesium deficiency favors the manifestation of low-grade chronic inflammation in obese subjects. From literature data, it is evident the participation of magnesium through biochemical and metabolic reactions in protecting against this metabolic disorder present in obesity.


HIV type 2 epidemic in Spain: challenges and missing opportunities.
: HIV type 2 (HIV-2) is a neglected virus despite estimates of 1-2 million people infected worldwide. HIV-2 is less efficiently transmitted than HIV-1 by sex and from mother to child. Although AIDS may develop in HIV-2 carriers, it takes longer than in HIV-1-infected patients. In contrast with HIV-1 infection, there is no global pandemic caused by HIV-2, as the virus is largely confined to West Africa. In a less extent and due to socioeconomic ties and wars, HIV-2 is prevalent in Portugal and its former colonies in Brazil, India, Mozambique and Angola. Globally, HIV-2 infections are steadily declining over time. A total of 338 cases of HIV-2 infection had been reported at the Spanish HIV-2 registry until December 2016, of whom 63% were men. Overall 72% were sub-Saharan Africans, whereas 16% were native Spaniards. Dual HIV-1 and HIV-2 coinfection was found in 9% of patients. Heterosexual contact was the most likely route of HIV-2 acquisition in more than 90% of cases. Roughly one-third presented with CD4 cell counts less than 200 cells/μl and/or AIDS clinical events. Plasma HIV-2 RNA was undetectable at baseline in 40% of patients. To date, one-third of HIV-2 carriers have received antiretroviral therapy, using integrase inhibitors 32 individuals. New diagnoses of HIV-2 in Spain have remained stable since 2010 with an average of 15 cases yearly. Illegal immigration from Northwestern African borders accounts for over 75% of new HIV-2 diagnoses. Given the relatively large community of West Africans already living in Spain and the continuous flux of immigration from endemic regions, HIV-2 infection either alone or as coinfection with HIV-1 should be excluded once in all HIV-seroreactive persons, especially when showing atypical HIV serological profiles, immunovirological disconnect (CD4 cell count loss despite undetectable HIV-1 viremia) and/or high epidemiological risks (birth in or sex partners from endemic regions).