Antimicrobial activity of <Emphasis Type="Italic">Alcaligenes sp</Emphasis>. HPC 1271 against multidrug resistant bacteria

Alcaligenes sp. HPC 1271 demonstrated antibacterial activity against multidrug resistant bacteria, Enterobacter sp., resistant to sulfamethoxazole, ampicillin, azithromycin, and tetracycline, as well as against Serratia sp. GMX1, resistant to the same antibiotics with the addition of netilmicin. The cell-free culture supernatant was analyzed for possible antibacterials by HPLC, and the active fraction was further identified by LC-MS. Results suggest the production of tunicamycin, a nucleoside antibiotic. The draft genome of this bacterial isolate was analyzed, and the 4.2 Mb sequence data revealed six secondary metabolite-producing clusters, identified using antiSMASH platform as ectoine, butyrolactone, phosphonate, terpene, polyketides, and nonribosomal peptide synthase (NRPS). Additionally, the draft genome demonstrated homology to the tunicamycin-producing gene cluster and also defined 30 ORFs linked to protein secretion that could also play a role in the antibacterial activity observed. Gene expression analysis demonstrated that both NRPS and dTDP-glucose 4,6-dehydratase gene clusters are functional and could be involved in antibacterial biosynthesis.     

Optimization of a preparative RP-HPLC method for isolation and purification of picrosides in <Emphasis Type="Italic">Picrorhiza kurroa</Emphasis>

Preparative reversed phase high pressure liquid chromatography (prep-RP-HPLC) coupled with photodiode array (PDA) and evaporative light scattering (ELSD) detectors was employed to isolate picrosides present in Picrorhiza kurroa Royle ex Benth. A binary gradient method (water and acetonitrile) was optimized on Water Spherisorb S10 ODS2 20 mm × 250 mm Semiprep Column with a 20 mL/min flow rate at ambient temperature with linear binary gradient conditions; at 0 min 15 % acetonitrile hold for 15 min; 15 to 22 % acetonitrile in next 2 min, hold for 13 min; 22 to 15 % acetonitrile in 5 min hold for 5 min to equilibrate column for next injection. The picroside-I and picroside-II fractions were 98.6 and 99.7 % pure with 13.9 mg and 9.8 mg yield per 200 mg of crude extract, respectively from mature dry rhizomes. Structures of isolated iridoids were confirmed with UV scan, 1H-NMR and direct infusion ESI-Q-TOF-MS/MS data.     

Quantification of IGF-1 Receptor May Be Useful in Diagnosing Polycythemia Vera–Suggestion to Be Added to Be One of the Minor Criterion

Endogenous erythroid colony (EEC) formation is one of the minor criteria for diagnosing polycythemia vera (PV) according to 2008 WHO diagnostic criteria. But EEC requires bone marrow aspiration and sophisticated laboratory procedures; therefore, practically it is rarely used to diagnose PV. Insulin-like growth factor 1 receptor (IGF-1R) was found to be constitutively phosphorylated and was responsible for the EEC formation in PV; therefore, we measured IGF-1R levels in the peripheral blood of 26 PV patients and compared them with those of 33 patients with secondary polycythemia and 29 normal controls. Among the PV patients, 16 were treated with only phlebotomy, 9 received hydroxyurea, and 1 was treated with ruxolinitinib. We found that PV patients treated with only phlebotomy had significantly higher IGF-1R levels than did those PV patients treated with hydroxyurea or ruxolinitinib. None of the secondary PV patients or normal controls had elevated IGR-1R levels, while 14 of 16 (87%) PV patients had significantly elevated IGF-1R levels. The new 2016 WHO has eliminated EEC as a minor criterion for diagnosing PV, but there are still some cases that cannot be definitively diagnosed by the current criteria. Therefore, we suggest that quantifying the IGF-1R level in peripheral blood by flow cytometry to replace EEC as the minor criterion for diagnosing PV.     

Efficacy of triapenthenol as a safener against metribuzin injury in soybean (Glycine max) cultivars

Triapenthenol or RSW 0411 (B(-cyclohexalmethylene)-gamma-(1,1-dimethylethyl)-1H-1,2,4-triazole-1-ethanol), a triazole plant growth regulator, applied preemergence as a separate broadcast application, protected tolerant and midtolerant soybean cultivars from metribuzin (4-amino-6-(1,1-dimethylethyl)-3-(methylthio)-1,2,4-triazin-5(4H)-one)-induced necrotic injury, and stabilized seedling fresh weight and dry weight loss to herbicide treatment. Soybean yields were not significantly reduced by triapenthenol treatment alone, but 1.12 kg ai/ha metribuzin and a 0.56 kg ai/ha triapenthenol plus 1.12 kg ai/ha metribuzin combination reduced crop yield averaged across cultivars.     

Phenol-mediated improved performance of active biomass for treatment of distillery wastewater

Activated sludge from a wastewater treatment plant consists of a consortium of microbes that utilize various organic molecules including persistent organic pollutants for their survival. Phenolic compounds and their derivatives along with dibenzofuran (DBF) are found as dominating pollutants in distillery waste. The acclimatization process leads to selective enrichment of the microbial community; and in this study, we report the acclimatizing effect of phenol on improving the treatment efficiency of two different distillery sludges—sludge from conventional aeration tanks (CAT), and from an extended aeration tank (EAT). The adaptation-dependent performance of activated biomass was studied by monitoring the increase in colony-forming units (CFUs) on mineral media and the utilization pattern for phenol (300×10³ and 530×10³ CFU for CAT and EAT sludge, respectively) and DBF (260×10³ and 430×10³ CFU for CAT and EAT sludge, respectively). The study showed that the acclimatization process remarkably improved the performance sludge for treatment of distillery wastewater. There was an improvement in chemical oxygen demand (COD) removal efficiency from 19% (unacclimatized sludge) to 31% in the case of acclimatized sludge (raw wastewater), which improved further to 82% and 87% with dilution of wastewater by 10 times (0.1×) and by 50 times (0.02×), respectively. Highest growth yields were observed with 0.1× wastewater (0.324 and 0.308 g g⁻¹ d⁻¹ for CAT and EAT sludges, respectively), while lower values are reported for the remaining two forms of wastewater. The study proposes that acclimatization step could be included as part of a treatment plant where the activated biomass could be intermittently metabolically charged by exposing it to selected molecules to increase treatment efficiency.     

Riboswitch regulation in cyanobacteria is independent of their habitat adaptations

Cyanobacteria are one of the ancient bacterial species occupying a variety of habitats with diverse metabolic preferences. RNA regulators like riboswitches play significant role in controlling the gene expression in prokaryotes. The taxonomic distribution of riboswitches suggests that they might be one of the oldest mechanisms of gene control system. In this paper, we analyzed the distribution of different riboswitch families in various cyanobacterial genomes. It was observed that only four riboswitch classes were abundant in cyanobacteria, B₁₂-element (Cob)/AdoCbl/AdoCbl-variant riboswitch being the most abundant. The analysis suggests that riboswitch mode of regulation is present in cyanobacterial species irrespective of their habitat types. A large number of unidentified genes regulated by riboswitches listed in this analysis indicate the wide range of targets for these riboswitch families. The analysis revealed a large number of genes regulated by riboswitches which may assist in elaborating the diversity among the cyanobacterial species.     

Genetic polymorphisms in carnitine palmitoyltransferase 1A gene are associated with variation in body composition and fasting lipid traits in Yup'ik Eskimos

Variants of carnitine palmitoyltransferase 1A (CPT1A), a key hepatic lipid oxidation enzyme, may influence how fatty acid oxidation contributes to obesity and metabolic outcomes. CPT1A is regulated by diet, suggesting interactions between gene variants and diet may influence outcomes. The objective of this study was to test the association of CPT1A variants with body composition and lipids, mediated by consumption of polyunsaturated fatty acids (PUFA). Obesity phenotypes and fasting lipids were measured in a cross-sectional sample of Yup'ik Eskimo individuals (n = 1141) from the Center of Alaska Native Health Research (CANHR) study. Twenty-eight tagging CPT1A SNPs were evaluated with outcomes of interest in regression models accounting for family structure. Several CPT1A polymorphisms were associated with HDL-cholesterol and obesity phenotypes. The P479L (rs80356779) variant was associated with all obesity-related traits and fasting HDL-cholesterol. Interestingly, the association of P479L with HDL-cholesterol was still significant after correcting for body mass index (BMI), percentage body fat (PBF), or waist circumference (WC). Our findings are consistent with the hypothesis that the L479 allele of the CPT1A P479L variant confers a selective advantage that is both cardioprotective (through increased HDL-cholesterol) and associated with reduced adiposity.     

Phylogenetic characterization of bacteria in the gut of house flies (Musca domestica L.)

House flies (Musca domestica L.) are cosmopolitan, ubiquitous, synanthropic insects that serve as mechanical or biological vectors for various microorganisms. To fully assess the role of house flies in the epidemiology of human diseases, it is essential to understand the diversity of microbiota harbored by natural fly populations. This study aimed to identify the diversity of house fly gut bacteria by both culture-dependent and culture-independent approaches. A total of 102 bacterial strains were isolated from the gut of 65 house flies collected from various public places including a garden, public park, garbage/dump area, public toilet, hospital, restaurant/canteen, mutton shop/market, and house/human habitation. Molecular phylogenetic analyses placed these isolates into 22 different genera. The majority of bacteria identified were known potential pathogens of the genera Klebsiella, Aeromonas, Shigella, Morganella, Providencia, and Staphylococcus. Culture-independent methods involved the construction of a 16S rRNA gene clone library, and sequence analyses supported culture recovery results. However, additional bacterial taxa not determined via culture recovery were revealed using this methodology and included members of the classes Alphaproteobacteria, Deltaproteobacteria, and the phylum Bacteroidetes. Here, we show that the house fly gut is an environmental reservoir for a vast number of bacterial species, which may have impacts on vector potential and pathogen transmission.