Molecular Targets for Diabetes Mellitus-associated Erectile Dysfunction

Protein expression profiles in rat corporal smooth muscle tissue were compared between animal models of streptozotocin-induced diabetes mellitus (STZ-DM) and agematched controls (AMCs) at 1 week and 2 months after induction of hyperglycemia with STZ treatment. At each time point, protein samples from four STZ-DM and four AMC rat corpora tissues were prepared independently and analyzed together across multiple quantitative two-dimensional gels using a pooled internal standard sample to quantify expression changes with statistical confidence. A total of 170 spots were differential expressed among the four experimental groups. A subsequent mass spectrometry analysis of the 170 spots identified a total of 57 unique proteins. Network analysis of these proteins using MetaCore™ suggested altered activity of transcriptional factors that are of too low abundance to be detected by the two-dimensional gel method. The proteins that were down-regulated with diabetes include isoforms of collagen that are precursors to fibril-forming collagen type 1; Hsp47, which assists and mediates the proper folding of procollagen; and several proteins whose abundance is controlled by sex hormones (e.g. CRP1 and A2U). On the other hand, proteins seen or predicted to be up-regulated include proteins involved in cell apoptosis (e.g. p53, 14-3-3-γ, Serpinf1, Cct4, Cct5, and Sepina3n), proteins that neutralize the biological activity of nerve growth factor (e.g. anti-NGF 30), and proteins involved in lipid metabolism (e.g. apoA-I and apoA-IV). Subsequent Western blot validation analysis of p53, 14-3-3-γ, and Hsp47 confirmed increased p53 and 14-3-3-γ and decreased Hsp47 levels in separate samples. According to the results from the Western blot analysis, Hsp47 protein showed a ∼3-fold decrease at 1 week and was virtually undetectable at 2 months in diabetic versus control. Taken together, our results identify novel candidate proteins playing a role in erectile dysfunction in diabetes resulting from STZ treatment

Proteomics Analysis Identifies Molecular Targets Related to Diabetes Mellitus-associated Bladder Dysfunction

Protein expression profiles in rat bladder smooth muscle were compared between animal models of streptozotocin-induced diabetes mellitus (STZ-DM) and age-matched controls at 1 week and 2 months after induction of hyper-glycemia with STZ treatment. At each time point, protein samples from four STZ-DM and four age-matched control rat bladder tissues were prepared independently and analyzed together across multiple DIGE gels using a pooled internal standard sample to quantify expression changes with statistical confidence. A total of 100 spots were determined to be significantly changing among the four experimental groups. A subsequent mass spectrometry analysis of the 100 spots identified a total of 56 unique proteins. Of the proteins identified by two-dimensional DIGE/MS, 10 exhibited significant changes 1 week after STZ-induced hyperglycemia, whereas the rest showed differential expression after 2 months. A network analysis of these proteins using MetaCore™ suggested induction of transcriptional factors that are too low to be detected by two-dimensional DIGE and identified an enriched cluster of down-regulated proteins that are involved in cell adhesion, cell shape control, and motility, including vinculin, intermediate filaments, Ppp2r1a, and extracellular matrix proteins. The proteins that were up-regulated include proteins involved in muscle contraction (e.g. MrIcb and Ly-GDI), in glycolysis (e.g. α-enolase and Taldo1), in mRNA processing (e.g. heterogeneous nuclear ribonucleoprotein A2/B1), in inflammatory response (e.g. S100A9, Annexin 1, and apoA-1), and in chromosome segregation and migration (e.g. Tuba1 and Vil2). Our results suggest that the development of diabetes-related complications in this model involves the down-regulation of structural and extracellular matrix proteins in smooth muscle that are essential for normal muscle contraction and relaxation but also induces proteins that are associated with cell proliferation and inflammation that may account for some of the functional deficits known to occur in diabetic complications of bladder

Trait based niche differentiation in tetrakas (Bernieridae) endemic to Madagascar: A multi-isotope approach

IntroductionTropical rainforest species interact with each other and their environment over a wide range of spatiotemporal scales. However, our understanding of resource partitioning and the mechanisms of avian species coexistence is largely restricted to subjective visual observations or acoustic monitoring. Therefore, the relative magnitudes of interspecific and intraspecific differences in resource use have remained difficult to quantify, particularly regarding different diets and habitat use. The eastern rainforest belt of Madagascar is inhabited by several species of insectivorous tetrakas belonging to an endemic bird family of Madagascar (Bernieridae). These species occupy similar habitats in the forest understory and are morphologically similar but because of likely differences (e.g., in foraging behaviors) we expect their foraging niches to be segregated allowing coexistence.MethodsWe examined the niche differentiation of four of these species: the Grey-crowned Tetraka (Xanthomixis cinereiceps), Long-billed Tetraka (Bernieria madagascariensis), Spectacled Tetraka (Xanthomixis zosterops), and White-throated Oxylabes (Oxylabes madagascariensis) in the Maromizaha rainforest in eastern Madagascar combining morphometry with stable carbon, nitrogen, and sulfur isotope ratios (δ13C, δ15N, and δ34S) from feathers.ResultsWe show considerable variation in isotopic niche positions, niche breadth and interspecific niche overlap. In two species, the Long-billed Tetraka and Spectacled Tetraka, we found an indication of sex-specific niche space, with males exhibiting a larger isotopic niche-area relative to females. Morphological traits of five species (including the Wedge-tailed Tetraka, Hartertula flavoviridis) coupled with stable isotope data provided explanations of patterns of niche overlap and isotopic position.DiscussionThe observed isotopic niche differences may be explained by differences in resource acquisition strategies that might be associated with specific morphological traits and spatial distribution. This may play an important role in niche differentiation among coexisting and phylogenetically closely related species

Polyamine stress at high pH in Escherichia coli K-12

BACKGROUND: Polyamines such as spermine and spermidine are required for growth of Escherichia coli; they interact with nucleic acids, and they bind to ribosomes. Polyamines block porins and decrease membrane permeability, activities that may protect cells in acid. At high concentrations, however, polyamines impair growth. They impair growth more severely at high pH, probably due to their increased uptake as membrane-permeant weak bases. The role of pH is critical in understanding polyamine stress. RESULTS: The effect of polyamines was tested on survival of Escherichia coli K-12 W3110 in extreme acid or base (pH conditions outside the growth range). At pH 2, 10 mM spermine increased survival by 2-fold, and putrescine increased survival by 30%. At pH 9.8, however, E. coli survival was decreased 100-fold by 10 mM spermine, putrescine, cadaverine, or spermidine. At pH 8.5, spermine decreased the growth rate substantially, whereas little effect was seen at pH 5.5. Spermidine required ten-fold higher concentrations to impair growth. On proteomic 2-D gels, spermine and spermidine caused differential expression of 31 different proteins. During log-phase growth at pH 7.0, 1 mM spermine induced eight proteins, including PykF, GlpK, SerS, DeaD, OmpC and OmpF. Proteins repressed included acetate-inducible enzymes (YfiD, Pta, Lpd) as well as RapA (HepA), and FabB. At pH 8.5, spermine induced additional proteins: TnaA, OmpA, YrdA and NanA (YhcJ) and also repressed 17 proteins. Four of the proteins that spermine induced (GlpK, OmpA, OmpF, TnaA) and five that were repressed (Lpd, Pta, SucB, TpiA, YfiD) show similar induction or repression, respectively, in base compared to acid. Most of these base stress proteins were also regulated by spermidine, but only at ten-fold higher concentration (10 mM) at high pH (pH 8.5). CONCLUSION: Polyamines increase survival in extreme acid, but decrease E. coli survival in extreme base. Growth inhibition by spermine and spermidine requires neutral or higher pH. At or above pH 7, spermine and spermidine regulate specific proteins, many of which are known to be regulated by base stress. High pH amplifies polyamine stress; and naturally occurring polyamines may play an important role in base stress

Feeding habits of culicine mosquitoes in the Cameroon lowland forests based on stable isotopes and blood meal analyses

Mosquito blood feeding behavior is a very significant component of pathogen transmission and determinant of disease epidemiology. Yet, knowledge of foraging ecology of mosquitoes often depends on the presence of undigested blood in the mosquito mid gut. Approximately 36 h after feeding, the blood meal is sufficiently digested to make identification by molecular techniques difficult, leaving a very narrow window in which these methods can be utilized. Here, we investigated the feeding habits of wild caught culicine mosquitoes from four genera, Aedes, Anopheles, Coquillettidia and Mansonia of the lowland rainforests of Cameroon based on the isotopic ratios of nitrogen (δ C appearing to be the best element to differentiate between mosquito species that fed on different host species. Isotopic analyses show that the different mosquito genera may be separated based on their diets, suggesting that linking stable isotope-based assays and DNA analysis may be a powerful new tool to investigate mosquito feeding ecology and the dynamics of vector-borne pathogens

Seasonal rainfall at long-term migratory staging sites is associated with altered carry-over effects in a Palearctic-African migratory bird.

BACKGROUND: An understanding of year-round habitat use is essential for determining how carry-over effects shape population dynamics in long-distance migratory songbirds. The recent discovery of long-term migratory staging sites in many species, prior to arrival at final wintering sites, adds complexity to efforts to decipher non-breeding habitat use and connections between sites. We investigated whether habitat conditions during migratory staging carry over to influence great reed warbler (Acrocephalus arundinaceus) body condition at final wintering sites in Zambia. We asked whether the presence/absence and strength of such carry-over effects were modified by contrasting rainfall conditions during 2 years. RESULTS: First, we found that individuals staging in a dry year had higher corticosterone (CORTf) and stable nitrogen isotope values (suggesting higher aridity) than birds staging in a wet year, indicating that regional weather affected staging conditions. Second, we found that carry-over effects from staging habitat conditions (measured via carbon and nitrogen isotopes) to final winter site body condition (measured via scaled mass index and β-hydroxybutyrate) were only present in a dry year, suggesting that environmental factors have consequences for the strength of carry-over effects. Our results also suggest that wet conditions at final winter sites may buffer the effects of poor staging conditions, at least in the short term, since individuals that staged in a dry year had higher scaled mass indices in Zambia than individuals that staged in a wet year. CONCLUSIONS: This study provides a first insight into the connections between long-term migratory staging sites and final wintering sites, and suggests that local environmental factors can modify the strength of carry-over effects for long-distance migratory birds.Gates Cambridge Trust, Natural Sciences and Engineering Research Council of Canada, Royal Society (Dorothy Hodgkin Fellowship), Biotechnology and Biological Sciences Research Council (David Phillips Fellowship, Grant ID: BB/J014109/1), DST-NRF Centre of Excellence at the FitzPatrick Institute, NERC (LSMSF Grant ID: EK206-16/12)This is the final version of the article. It first appeared from BioMed Central via http://dx.doi.org/10.1186/s12898-016-0096-

Oxygen limitation modulates pH regulation of catabolism and hydrogenases, multidrug transporters, and envelope composition in Escherichia coli K-12

BACKGROUND: In Escherichia coli, pH regulates genes for amino-acid and sugar catabolism, electron transport, oxidative stress, periplasmic and envelope proteins. Many pH-dependent genes are co-regulated by anaerobiosis, but the overall intersection of pH stress and oxygen limitation has not been investigated. RESULTS: The pH dependence of gene expression was analyzed in oxygen-limited cultures of E. coli K-12 strain W3110. E. coli K-12 strain W3110 was cultured in closed tubes containing LBK broth buffered at pH 5.7, pH 7.0, and pH 8.5. Affymetrix array hybridization revealed pH-dependent expression of 1,384 genes and 610 intergenic regions. A core group of 251 genes showed pH responses similar to those in a previous study of cultures grown with aeration. The highly acid-induced gene yagU was shown to be required for extreme-acid resistance (survival at pH 2). Acid also up-regulated fimbriae (fimAC), periplasmic chaperones (hdeAB), cyclopropane fatty acid synthase (cfa), and the "constitutive" Na+/H+ antiporter (nhaB). Base up-regulated core genes for maltodextrin transport (lamB, mal), ATP synthase (atp), and DNA repair (recA, mutL). Other genes showed opposite pH responses with or without aeration, for example ETS components (cyo,nuo, sdh) and hydrogenases (hya, hyb, hyc, hyf, hyp). A hypF strain lacking all hydrogenase activity showed loss of extreme-acid resistance. Under oxygen limitation only, acid down-regulated ribosome synthesis (rpl,rpm, rps). Acid up-regulated the catabolism of sugar derivatives whose fermentation minimized acid production (gnd, gnt, srl), and also a cluster of 13 genes in the gadA region. Acid up-regulated drug transporters (mdtEF, mdtL), but down-regulated penicillin-binding proteins (dacACD, mreBC). Intergenic regions containing regulatory sRNAs were up-regulated by acid (ryeA, csrB, gadY, rybC). CONCLUSION: pH regulates a core set of genes independently of oxygen, including yagU, fimbriae, periplasmic chaperones, and nhaB. Under oxygen limitation, however, pH regulation is reversed for genes encoding electron transport components and hydrogenases. Extreme-acid resistance requires yagU and hydrogenase production. Ribosome synthesis is down-regulated at low pH under oxygen limitation, possibly due to the restricted energy yield of catabolism. Under oxygen limitation, pH regulates metabolism and transport so as to maximize alternative catabolic options while minimizing acidification or alkalinization of the cytoplasm

Plasma metabolomics and clinical predictors of survival differences in COPD patients

Background: Plasma metabolomics profile (PMP) in COPD has been associated with clinical characteristics, but PMP''s relationship to survival has not been reported. We determined PMP differences between patients with COPD who died an average of 2 years after enrollment (Non-survivors, NS) compared to those who survived (S) and also with age matched controls (C). Methods: We studied prospectively 90 patients with severe COPD and 30 controls. NS were divided in discovery and validation cohorts (30 patients each) and the results compared to the PMP of 30 S and C. All participants completed lung function tests, dyspnea scores, quality of life, exercise capacity, BODE index, and plasma metabolomics by liquid and gas chromatography/mass spectometry (LC/MS, LC/MS2, GC/MS). Statistically, we used Random Forest Analysis (RFA) and Support Vector Machine (SVM) to determine metabolites that differentiated the 3 groups and compared the ability of metabolites vs. clinical characteristics to classify patients into survivors and non-survivors. Results: There were 79 metabolites statistically different between S and NS [p < 0.05 and false discovery rate (q value) < 0.1]. RFA and SVM classification of COPD survivors and non-survivors had a predicted accuracy of 74 and 85% respectively. Elevation of tricyclic acid cycle intermediates branched amino acids depletion and increase in lactate, fructose and xylonate showed the most relevant differences between S vs. NS suggesting alteration in mitochondrial oxidative energy generation. PMP had similar predictive power for risk of death as information provided by clinical characteristics. Conclusions: A plasma metabolomic profile characterized by an oxidative energy production difference between survivors and non-survivors was observed in COPD patients 2 years before death

Outcome of preterm twins compared to preterm singleton neonates, a multicenter prospective observational study in Ethiopia

Background: In recent decades there has been a major increase in multiple birth rates, and the rate of twining vary from 6-9 per thousand life births to 20 per thousand live births across differ-ent areas of the world. Many studies have demonstrated higher neonatal and perinatal mortality and morbidity rates in twin deliveries compared to singleton births. This study was aimed to com-pare the outcomes of preterm twins and preterm singletons.Methods: A prospective, observational multicenter study was conducted from July 2016 to May 2018 in five tertiary hospitals in Ethiopia. All preterm, liveborn infants born at or transferred at less than 7 days of life to one of the study hospitals with an estimated gestational age below 37 weeks were included.Results: A total of 3,703 preterm neonates admitted to participating neonatal intensive care units were included in the study, of which 1171(31.6%) were twins. After adjusting for birth weight and gestational age, the mortality rate for preterm singletons of 31.0% was higher than the mortality rate for preterm twins of 24.8%, which was statistically significant (p-value = 0.001), OR of 1.37 (95% CI: 1.15 to 1.64). The study also identified an inverse relationship between birth weight and gestational age, and mortality. Male singletons were more likely to die than male twins (440 (32.4%) vs. 141 (23.4%); AOR 1.56 (95% CI: 1.22, 1.99); p=0.001)Conclusion: Our study showed that the mortality of a singleton preterm infant was significantly higher than the mortality of a preterm twin infan