Galleria mellonella Infection Model Demonstrates High Lethality of ST69 and ST127 Uropathogenic E. coli

Galleria mellonella larvae are an alternative in vivo model for investigating bacterial pathogenicity. Here, we examined the pathogenicity of 71 isolates from five leading uropathogenic E. coli (UPEC) lineages using G. mellonella larvae. Larvae were challenged with a range of inoculum doses to determine the 50% lethal dose (LD₅₀) and for analysis of survival outcome using Kaplan-Meier plots. Virulence was correlated with carriage of a panel of 29 virulence factors (VF). Larvae inoculated with ST69 and ST127 isolates (10⁴ colony-forming units/larvae) showed significantly higher mortality rates than those infected with ST73, ST95 and ST131 isolates, killing 50% of the larvae within 24 hours. Interestingly, ST131 isolates were the least virulent. We observed that ST127 isolates are significantly associated with a higher VF-score than isolates of all other STs tested (P≤0.0001), including ST69 (P<0.02), but one ST127 isolate (strain EC18) was avirulent. Comparative genomic analyses with virulent ST127 strains revealed an IS1 mediated deletion in the O-antigen cluster in strain EC18, which is likely to explain the lack of virulence in the larvae infection model. Virulence in the larvae was not correlated with serotype or phylogenetic group. This study illustrates that G. mellonella are an excellent tool for investigation of the virulence of UPEC strains. The findings also support our suggestion that the incidence of ST127 strains should be monitored, as these isolates have not yet been widely reported, but they clearly have a pathogenic potential greater than that of more widely recognised clones, including ST73, ST95 or ST131.     

Population size and habitat preference of the Omo River guereza (Colobus guereza guereza) in a multi-habitat matrix in the central highlands of Ethiopia

Primates - Given the current rate of habitat degradation and loss in the tropics, data on primate population densities and habitat use are indispensable for assessing conservation status and...     

Geographic distribution of microsatellite alleles in geladas (Primates,Cercopithecidae): Evidence for three evolutionary units

The subspecific taxonomy and distribution of geladas (Theropithecus gelada Rüppell, 1835) remains uncertain. Recent molecular studies based on mitochondrial sequence data revealed a geographically structured, three-deme population, suggesting that there are three evolutionary units of geladas. However, mitochondrial distributions do not always recover population relationships, particularly in taxa with a complex history of isolation and gene flow. We therefore analysed the nuclear genetic population structure of the global gelada population based on 20 microsatellite loci in 43 samples from across its geographic range. F_ST values, a STRUCTURE analysis and a principal coordinate analysis (PCoA) confirmed the three-deme population structure corresponding to the mitochondrial population structure. Therefore, our analyses provide additional support for three evolutionary units in geladas, corresponding to (a) a northern (north of Lake Tana, primarily in the Simien Mountains, previously classified as Theropithecus gelada gelada Rüppell, 1835), (b) a central (between Addis Ababa and the highlands east of Lake Tana, previously classified as Theropithecus gelada obscurus Heuglin, 1863) and (c) a southern (south of the Rift Valley, previously tentatively classified as Theropithecus gelada arsi Shotake et al., 2016, Anthropological Science, 124, 157) population. These results pave the way for future conservation decisions and highlight that the gelada population boundaries need more fine-grained genetic sampling and phenotypic analyses, in particular for their taxonomic ranking.     

TCF7L2 gene polymorphism as a risk for type 2 diabetes mellitus and diabetic microvascular complications

Molecular Biology Reports - Type 2 Diabetes Mellitus (T2DM) is a chronic metabolic condition with various genetics and environmental influences that affects the capacity of the body to produce or...     

Screening of promising molecules against MurG as drug target in multi-drug-resistant-Acinetobacter baumannii - insights from comparative protein modeling,molecular docking and molecular dynamics simulation

Abstract

The UDP-N-acetylglucosamine-N-acetylmuramyl-(pentapeptide) pyrophosphoryl-undecaprenol N-acetylglucosamine transferase (MurG) is located in plasma membrane which plays a crucial role for peptidoglycan biosynthesis in Gram-negative bacteria. Recently, this protein is considered as an important and unique drug target in Acinetobacter baumannii since it plays a key role during the synthesis of peptidoglycan as well as which is not found in Homo sapiens. In this study, initially we performed comparative protein modeling approach to predict the three-dimensional model of MurG based on crystal structure of UDP-N-acetylglucosamine-N-acetylmuramyl-(pentapeptide) pyrophosphoryl-undecaprenol N-acetylglucosamine transferase (PDB ID: 1F0K) from E.coli K12. MurG model has two important functional domains located in N and C- terminus which are separated by a deep cleft. Active site residues are located between two domains and they are Gly20, Arg170, Gly200, Ser201, Gln227, Phe254, Leu275, Thr276, and Glu279 which play essential role for the function of MurG. In order to inhibit the function of MurG, we employed the High Throughput Virtual Screening (HTVS) and docking techniques to identify the promising molecules which will further subjected into screening for computing their drug like and pharmacokinetic properties. From the HTVS, we identified 5279 molecules, among these, 12 were passed the drug-like and pharmacokinetic screening analysis. Based on the interaction analysis in terms of binding affinity, inhibition constant and intermolecular interactions, we selected four molecules for further MD simulation to understand the structural stability of protein-ligand complexes. All the analysis of MD simulation suggested that ZINC09186673 and ZINC09956120 are identified as most promising putative inhibitors for MurG protein in A. baumannii.

Communicated by Ramaswamy H. Sarma     

The alien flora of Sudan and South Sudan: taxonomic and biogeographical composition

Biological Invasions - Studies on plant invasions depend on local and regional checklists of the alien flora. However, global overview studies have shown that some regions, including many African...     

Metabolomic Method: UPLC-q-ToF Polar and Non-Polar Metabolites in the Healthy Rat Cerebellum Using an In-Vial Dual Extraction

Unbiased metabolomic analysis of biological samples is a powerful and increasingly commonly utilised tool, especially for the analysis of bio-fluids to identify candidate biomarkers. To date however only a small number of metabolomic studies have been applied to studying the metabolite composition of tissue samples, this is due, in part to a number of technical challenges including scarcity of material and difficulty in extracting metabolites. The aim of this study was to develop a method for maximising the biological information obtained from small tissue samples by optimising sample preparation, LC-MS analysis and metabolite identification. Here we describe an in-vial dual extraction (IVDE) method, with reversed phase and hydrophilic liquid interaction chromatography (HILIC) which reproducibly measured over 4,000 metabolite features from as little as 3mg of brain tissue. The aqueous phase was analysed in positive and negative modes following HILIC separation in which 2,838 metabolite features were consistently measured including amino acids, sugars and purine bases. The non-aqueous phase was also analysed in positive and negative modes following reversed phase separation gradients respectively from which 1,183 metabolite features were consistently measured representing metabolites such as phosphatidylcholines, sphingolipids and triacylglycerides. The described metabolomics method includes a database for 200 metabolites, retention time, mass and relative intensity, and presents the basal metabolite composition for brain tissue in the healthy rat cerebellum.     

Assessment of Water Quality of Soan River using Physicochemical Parameters and Aquatic Insects Diversity

Biology Bulletin - In the present study, the effect of pollutants produced by human dwelling, agricultural and industrial activities on aquatic invertebrate’s communities of Soan River,...     

Fermentation of Barley by Using Saccharomyces cerevisiae: Examination of Barley as a Feedstock for Bioethanol Production and Value-Added Products

The objective of this study was to examine the ethanol yield potential of three barley varieties (Xena, Bold, and Fibar) in comparison to two benchmarks, corn and wheat. Very high gravity (VHG; 30% solids) fermentations using both conventional and Stargen 001 enzymes for starch hydrolysis were carried out as simultaneous saccharification and fermentation. The grains and their corresponding dried distiller''s grain with solubles (DDGS) were also analyzed for nutritional and value-added characteristics. A VHG traditional fermentation approach utilizing jet-cooking fermentation revealed that both dehulled Bold and Xena barley produced ethanol concentrations higher than that produced by wheat (12.3, 12.2, and 11.9%, respectively) but lower than that produced by corn (13.8%). VHG-modified Stargen-based fermentation of dehulled Bold barley demonstrated comparable performance (14.3% ethanol) relative to that of corn (14.5%) and wheat (13.3%). Several important components were found to survive fermentation and were concentrated in DDGS. The highest yield of phenolics was detected in the DDGS (modified Stargen 001, 20% solids) of Xena (14.6 mg of gallic acid/g) and Bold (15.0 mg of gallic acid/g) when the hull was not removed before fermentation. The highest concentration of sterols in DDGS from barley was found in Xena (3.9 mg/g) when the hull was included. The DDGS recovered from corn had the highest concentration of fatty acids (72.6 and 77.5 mg/g). The DDGS recovered from VHG jet-cooking fermentations of Fibar, dehulled Bold, and corn demonstrated similar levels of tocopherols and tocotrienols. Corn DDGS was highest in crude fat but was lowest in crude protein and in vitro energy digestibility. Wheat DDGS was highest in crude protein content, similar to previous studies. The barley DDGS was the highest in in vitro energy digestibility.The growing need for energy independence and proposed renewable fuels has led recently to a major expansion of fuel ethanol production. In North America, this activity primarily uses corn as a feedstock. The need to find other cost-effective and efficient grains for ethanol production has increased in significance. Cereal grains are high in starch and are currently being utilized for ethanol production (26, 41). To ensure long-term viability of the industry, fermentation strategies that focus on holistic utilization of the feedstock that maximize value addition will increase in importance. The focus of industry is slowly moving from biorefineries that anticipate subsidy and government policy to integrated biorefineries that produce multiple products. Multiple product streams and integrated by-product management are thought to ensure better financial stability and opportunities for diversified income streams.Barley is a potential candidate for industrial ethanol production (10) since its ethanol yield is comparable to that of wheat but below that of American corn, which is currently the preferred industrial feedstock. Barley contains on average 63 to 65% starch, 8 to 13% protein, 2 to 3% fat, 1 to 1.5% soluble gums, 8 to 10% hemicellulose, ca. 2.9% lignin, and 2 to 2.5% ash (15, 27). Barley also contains a hull that could be fermented using cellulolytic enzymes, providing opportunities for integrated biorefineries that utilize more feedstocks than corn. Potential coproducts of ethanol production from barley include protein, fiber, fatty acids, tocopherols, and tocotrienols (40). The nutritional value of barley, based on amino acid content, is greater than that for corn and is not significantly affected by the fermentation process (40). A range of nutraceutical and functional food products, as well as amylase, amylase inhibitors, β-amylase, and oxalate oxidase, are found in barley grains and may have potential for extraction and commercial applications (6, 22, 33). Hull-less barley lines, high in both protein (particularly lysine) and starch, and low in fiber, have recently been developed (11, 14, 32). Since starch recovery and thus ethanol yields are lower for barley than corn, coproduct recovery becomes even more essential for profitability (43).Enzymes used for the pretreatment of grains prior to fermentation have traditionally been α-amylases and glucoamylases. The α-amylase decreases the viscosity of the mash (25) and performs the liquefaction of the pretreatment process. The liquefaction step is typically done at high temperatures of 100 to 120°C (38) with direct steam injection (jet-cooking). The α-amylase action serves to break starch at α-(1,4)-glucosidic bonds, producing smaller dextrin chains. During the saccharification step of the pretreatment, the dextrins produced by α-amylase are then acted on by glucoamylase. This conventional method has a considerable economic drawback, because the mash must undergo a cooking step prior to fermentation. Many industrial ethanol producers use jet-cooking to raise the mash temperature to 100 to 120°C. Because of this temperature requirement, the conventional process uses a large amount of energy to produce ethanol.Recently, a new line of cold starch hydrolyzing enzymes was developed. An example of these enzymes is Stargen 001, which is referred to as a raw starch hydrolyzing enzyme because starch is hydrolyzed to fermentable sugars while the temperature remains at or below a temperature of 48°C (38). Stargen 001 replaces the liquefaction and saccharification steps performed by conventional digestion enzymes (i.e., α-amylase and glucoamylase) and releases free glucose and other fermentable sugars for use by yeast cells. Stargen 001 is a cocktail of modified α-amylase and glucoamylase enzymes that work together to convert starch into dextrins, followed by the hydrolysis of dextrins to fermentable sugars (37, 38). With the absence of a cooking stage in the cold hydrolysis method, the potential exists that the dried distiller''s grain plus solubles (DDGS) produced by fermentation would have less damage so that the proteins contained in the DDGS could be of more value (18).The objectives of the present study were to examine the ethanol yield potentials of three barley varieties (Xena, Bold, and Fibar) and two benchmark grains (Pioneer Hi-Bred corn and CPS wheat) using conventional (jet-cooking) and cold starch hydrolysis with Stargen 001. In addition, dehulling was tested for the potential to increase ethanol yields, because hull does not contain fermentable starch; both hulled and dehulled mashes were studied where possible. The grains and their corresponding DDGS were analyzed for nutritional value and the presence of potential value-added products such as fatty acids, tocopherols, tocotrienols, sterols, and polyphenols.