Gut integrity and duodenal enteropathogen burden in undernourished children with environmental enteric dysfunction

Environmental enteric dysfunction (EED) is a subclinical condition of intestinal inflammation, barrier dysfunction and malabsorption associated with growth faltering in children living in poverty. This study explores association of altered duodenal permeability (lactulose, rhamnose and their ratio) with higher burden of enteropathogen in the duodenal aspirate, altered histopathological findings and higher morbidity (diarrhea) that is collectively associated with linear growth faltering in children living in EED endemic setting. In a longitudinal birth cohort, 51 controls (WHZ > 0, HAZ > −1.0) and 63 cases (WHZ< -2.0, refractory to nutritional intervention) were recruited. Anthropometry and morbidity were recorded on monthly bases up to 24 months of age. Dual sugar assay of urine collected after oral administration of lactulose and rhamnose was assessed in 96 children from both the groups. Duodenal histopathology (n = 63) and enteropathogen analysis of aspirate via Taqman array card (n = 60) was assessed in only cases. Giardia was the most frequent pathogen and was associated with raised L:R ratio (p = 0.068). Gastric microscopy was more sensitive than duodenal aspirate in H. pylori detection. Microscopically confirmed H. pylori negatively correlated with HAZ at 24 months (r = −0.313, p = 0.013). Regarding histopathological parameters, goblet cell reduction significantly correlated with decline in dual sugar excretion (p< 0.05). Between cases and controls, there were no significant differences in the median (25^th, 75^th percentile) of urinary concentrations (μg/ml) of lactulose [27.0 (11.50, 59.50) for cases vs. 38.0 (12.0, 61.0) for controls], rhamnose [66.0 (28.0, 178.0) vs. 86.5 (29.5, 190.5)] and L:R ratio [0.47 (0.24, 0.90) vs. 0.51 (0.31, 0.71)] respectively. In multivariable regression model, 31% of variability in HAZ at 24 months of age among cases and controls was explained by final model including dual sugars. In conclusion, enteropathogen burden is associated with altered histopathological features and intestinal permeability. In cases and controls living in settings of endemic enteropathy, intestinal permeability test may predict linear growth. However, for adoption as a screening tool for EED, further validation is required due to its complex intestinal pathophysiology.     

Evaluation the Properties of Polycaprolactone/Fluorapatite Nano-biocomposite

In this study,nano-biocomposites of polycaprolactone(PCL)as the matrix and different amounts of nanofluorapatite(nFA)(0,10,20 and 30 wt.％)as the reinforcement w...     

Quantifying dynamic mechanical properties of human placenta tissue using optimization techniques with specimen-specific finite-element models

Motor-vehicle crashes are the leading cause of fetal deaths resulting from maternal trauma in the United States, and placental abruption is the most common cause of these deaths. To minimize this injury, new assessment tools, such as crash-test dummies and computational models of pregnant women, are needed to evaluate vehicle restraint systems with respect to reducing the risk of placental abruption. Developing these models requires accurate material properties for tissues in the pregnant abdomen under dynamic loading conditions that can occur in crashes. A method has been developed for determining dynamic material properties of human soft tissues that combines results from uniaxial tensile tests, specimen-specific finite-element models based on laser scans that accurately capture non-uniform tissue-specimen geometry, and optimization techniques. The current study applies this method to characterizing material properties of placental tissue. For 21 placenta specimens tested at a strain rate of 12/s, the mean failure strain is 0.472±0.097 and the mean failure stress is 34.80±12.62 kPa. A first-order Ogden material model with ground-state shear modulus (μ) of 23.97±5.52 kPa and exponent (α₁) of 3.66±1.90 best fits the test results. The new method provides a nearly 40% error reduction (p<0.001) compared to traditional curve-fitting methods by considering detailed specimen geometry, loading conditions, and dynamic effects from high-speed loading. The proposed method can be applied to determine mechanical properties of other soft biological tissues.     

Mating tactics of male feral goats (Capra hircus): risks and benefits

Intense competition between males for reproduction has led to the evolution of alternative mating tactics (AMTs). Feral goat males usually use a tactic called tending, in which they defend oestrous females from other males. Males may also use a second mating tactic called coursing, in which they gain access to oestrous females by disturbing a tending pair. Herein, we examine estimated mating success (EMS) and risks of using these tactics. Tending was only used by mature (≥4 years old), higher-ranking males and accounted for 75% of EMS. Coursing was used by males of all ages and dominance ranks, and accounted for 25% of EMS. Using coursing, male kids achieved 8% of EMS. Mature males achieved 92% of EMS. Both age and dominance rank were related to EMS, but age was not important after its relationship with dominance was controlled. Tending bouts were, on average, ca. 30 min long, while coursing bouts only averaged ca. 2 min. Males were more likely to suffer a butt while coursing than while tending, and formerly tending males were responsible for most butts. Kids that coursed had the highest risk of being butted. In most AMTs, there are reductions in the risks in relation to low fitness benefits. However, we found that the risks of butts during coursing were high, while our evidence suggests that the EMS was probably low. Nevertheless, the existence of an effective AMT in male feral goats may have an important influence on the intensity of sexual selection and the effective population size.     

Isolation and characterization of novel phenotype CHO cell mutants defective in peroxisome assembly, using ICR191 as a potent mutagenic agent

To elucidate molecular and cellular mechanisms of peroxisome biogenesis, we have isolated Chinese hamster ovary (CHO) cell mutants defective in peroxisome biogenesis by making use of enhanced green fluorescent protein (EGFP) and a frameshift-inducing mutagen ICR191. CHO-TKa cells stably expressing Pex2p were transformed with a cDNA encoding EGFP fused with peroxisomal targeting signal type 2 (PTS2-EGFP), termed Tka/EG2. TKa/EG2 cells were mutagenized with ICR191 and cultured in the presence of P9OH (9-(1'-pyrene) nonanol) followed by an exposure to UV. P9OH/UV-resistant and morphologically peroxisome-deficient mutant cells were isolated by directly observing cytosolic localization of EGFP, without cell staining. By a combination of cell-fusion and PEX transfection, we determined complementation groups (CGs) of 16 cell mutants isolated here. The mutants were classified into five CGs, including pex2, pex3, pex5, pex6, and pex7 cell mutants. In contrast to typical pex6 mutants with the impaired import of both PTS1- and PTS2-proteins, two clones, ZPEG236 and ZPEG244, showed a distinct, novel phenotype where PTS1-protein import was normal despite the abrogated PTS2 import. Dysfunction of Pex3p in pex3 ZPEG 238 was due to one base (G) insertion in the codon for Asn7 resulting in a frameshift, thereby inducing a distinct 31 amino-acid sequence and a termination. pex2 ZPEG239 showed a mutation in codon GAG for Glu(201) to a nonsense mutation, TAG. Thus, the method developed here using ICR191 could be useful for isolation of further novel cell mutants impaired in peroxisome biogenesis.     

Vibrio harveyi flavin reductase--luciferase fusion protein mimics a single-component bifunctional monooxygenase

Vibrio harveyi luciferase and flavin reductase FRP are, together, a two-component monooxygenase couple. The reduced flavin mononucleotide (FMNH2) generated by FRP must be supplied, through either free diffusion or direct transfer, to luciferase as a substrate. In contrast, single-component bifunctional monooxygenases each contains a bound flavin cofactor and does not require any flavin addition to facilitate catalysis. In this study, we generated and characterized a novel fusion enzyme, FRP-alphabeta, in which FRP was fused to the luciferase alpha subunit. Both FRP and luciferase within FRP-alphabeta were catalytically active. Kinetic properties characteristic of a direct transfer of FMNH2 cofactor from FRP to luciferase in a FRP:luciferase noncovalent complex were retained by FRP-alphabeta. At submicromolar levels, FRP-alphabeta was significantly more active than an equal molar mixture of FRP and luciferase in coupled bioluminescence without FMN addition. Importantly, FRP-alphabeta gave a higher total quantum output without than with exogenously added FMN. Moreover, effects of increasing concentrations of oxygen on light intensity were investigated using sub-micromolar enzymes, and results indicated that the bioluminescence produced by FRP-alphabeta without added flavin was derived from direct transfer of reduced flavin whereas bioluminescence from a mixture of FRP and luciferase with or without exogenously added flavin relied on free-diffusing reduced flavin. Therefore, the overall catalytic reaction of FRP-alphabeta without any FMN addition closely mimics that of a single-component bifunctional monooxygenase. This fusion enzyme approach could be useful to other two-component monooxygenases in enhancing the enzyme efficiencies under conditions hindering reduced flavin delivery. Other potential utilities of this approach are discussed.     

Habitat suitability and connectivity implications for the conservation of the Persian leopard along the Iran–Iraq border

Habitat fragmentation has major negative impacts on wildlife populations, and the connectivity could reduce these negative impacts. This study was conducted to assess habitat suitability and structural connectivity of the Persian leopard along the Iran–Iraq border (i.e., the Zagros Mountains) and compare the situation of identified core habitats and connectivity with existing conservation areas (CAs). An ensemble modeling approach resulting from five models was used to predict habitat suitability. To identify core habitats and corridors along the Iran–Iraq border, factorial least‐cost path analyses were applied. The results revealed that topographic roughness, distance to CAs, annual precipitation, vegetation/cropland density, and distance to rivers were the most influential variables for predicting the occurrence of the Persian leopard in the study area. By an estimated dispersal distance of 82 km (suggested by previous studies), three core habitats were identified (two cores in Iran and one core in Iraq). The largest cores were located in the south and the center of the study area, which had the highest connectivity priorities. The connectivity from these cores was maintained to the core within the Iraqi side. Only about one‐fifth of detected core habitats and relative corridors were protected by CAs in the study area. Detected core habitats and connectivity areas in this study could be an appropriate road map to accomplish the CAs network along the Iran–Iraq border regarding Persian leopard conservation. Establishing transboundary CAs, particularly in the core habitat located in the center of the study area, is strongly recommended to conserve existing large carnivores, including the Persian leopard.     

Multiple gene genealogies and AFLPs suggest cryptic speciation and long-distance dispersal in the basidiomycete Serpula himantioides (Boletales)

Serpula himantioides (Boletales, Basidiomycota) produces thin resupinate basidiocarps on dead coniferous wood worldwide and causes damage in buildings as well. In this study, we present evidence for the existence of at least three phylogenetically defined cryptic species (referred to as Sib I-III) within the morphospecies S. himantioides, a conclusion based on analyses of sequence data from four DNA regions and amplified fragment length polymorphisms (AFLPS). A low degree of shared sequence polymorphisms was observed among the three lineages indicating a long-lasting separation. The AFLPs revealed two additional subgroups within Sib III. Results from mating studies were consistent with the molecular data. In Sib III, no correspondence between genetic and geographical distance was observed among isolates worldwide, presumably reflecting recent dispersal events. Our results indicate that at least two of the lineages (Sib II and Sib III) have wide sympatric distributions. A population genetic analysis of Sib III isolates, scoring sequence polymorphisms as codominant SNP markers, indicates that panmictic conditions exist in the Sib III group. This study supports the view that cryptic speciation is a common phenomenon in basidiomycete fungi and that phylogenetic species recognition can be a powerful inference to detect cryptic species. Furthermore, this study shows that AFLP data are a valuable supplement to DNA sequence data in that they may detect a finer level of genetic variation.     

Hexahydro-pyrrolo- and hexahydro-1H-pyrido[1,2-b]pyridazin-2-ones as potent inhibitors of HCV NS5B polymerase

Hexahydro-pyrrolo- and hexahydro-1H-pyrido[1,2-b]pyridazin-2-one analogs were discovered as a novel class of inhibitors of genotype 1 HCV NS5B polymerase. Among these, compound 4c displayed potent inhibitory activities in biochemical and replicon assays (IC(50) (1b) <10 nM; EC(50) (1b)=34 nM) as well as good stability towards human liver microsomes (HLM t(1/2) =59 min).     

Homology Modeling of Hemagglutinin/Protease [HA/P (vibriolysin)] from <Emphasis Type="Italic">Vibrio Cholerae</Emphasis>: Sequence Comparision,Residue Interactions and Molecular Mechanism

Vibrio cholerae produces a zinc-containing and calcium-stabilized soluble hemagglutinin/protease, which has been earlier shown to have the ability to cleave several physiologically important substrates including mucin, fibronectin and lactoferin. This study presents homology modeling of hemagglutinin/protease (vibriolysin) from Vibrio cholerae in the presence of inhibitor HPI [N-(1-carboxy-3-phenylpropyl)-phenylalanyl-alpha-aspargine]. The 3D structure was predicted based on its sequence homology with Pseudomonas aeruginosa elastase (PAE). Comparison of the 3D structures of PAE and HA/P reveals a remarkable similarity having a conserved α + β domain. The inhibitor shows similar binding features as seen in other metalloproteases of M4 peptidase family. The study also highlights the key catalytic residues as well as the residues at the S₁ and binding sub-sites. The similarities between the two proteins provide support for the hypothesis that the two enzymes have similar three-dimensional structures and a common mechanism of action. The fact that both enzymes are secreted as zinc-containing proteases, led us to further hypothesize that they may play similar role in pathogenesis.