<i>In silico</i> and bio assay of juvenile hormone analogs as an insect growth regulator against <i>Galleria mellonella</i> (wax moth) – Part I

<p>Juvenile hormone (JH) analogs are nowadays in use to control harmful pests. In order to develop new bioactive molecules as potential pesticides, we have incorporated different active structural features like sulfonamide, aromatic rings, amide group, and amino acid moiety to the base structure. We have screened a series of designed novel JH analogs against JH receptor protein (jhbpGm-2RCK) of <i>Galleria mellonella</i> in comparison to commercial insect growth regulators (IGRs) – Pyriproxyfen (T₁) and Fenoxycarb (T₂). All analogs exhibit the binding energy profile comparable to commercial IGRs. Based upon these results, a series of sulfonamide-based JHAs (T₃–T₈) as IGRs have been synthesized and characterized. Further, the efficacy of synthesized analogs (T₃–T₈) and commercial IGRs (Pyriproxyfen and Fenoxycarb) has been assessed against fourth instars larvae of <i>G. mellonella</i> under the laboratory conditions. LC₅₀ values of all the analogs (T₁–T₈) against the fourth instars larvae were 9.99, 10.12, 24.76, 30.73, 38.45, 34.15, 34.14, 19.48 ppm and the LC₉₀ 153.27, 131.69, 112.15, 191.46, 427.02, 167.13, 217.10, 172.00 ppm, respectively. Among these analogs, N-(1-isopropyl-2-oxo-3-aza-3-N-ethyl-pentanyl)-p-toluene sulfonamide (T₈) and N-(1-isopropyl-2-oxo-3-aza-3-N-ethyl-pentanyl) benzene sulfonamide (T₇) exhibited the good pest larval mortality at different exposure periods (in hours) and different concentrations (in ppm) in comparison to in use IGRs- T₁ and T₂. Bio assay results are supported by docking at higher concentration. The present investigation clearly exhibits that analog T₈ could serve as a potential IGR in comparison to in use IGRs (T₁ and T₂). The results are promising and provide new array of synthetic chemicals that may be utilized as IGRs.</p

Zinc Finger Domain of the PRDM9 Gene on Chromosome 1 Exhibits High Diversity in Ruminants but Its Paralog PRDM7 Contains Multiple Disruptive Mutations

<div><p>PRDM9 is the sole hybrid sterility gene identified so far in vertebrates. PRDM9 gene encodes a protein with an immensely variable zinc-finger (ZF) domain that determines the site of meiotic recombination hotspots genome-wide. In this study, the terminal ZF domain of PRDM9 on bovine chromosome 1 and its paralog on chromosome 22 were characterized in 225 samples from five ruminant species (cattle, yak, mithun, sheep and goat). We found extraordinary variation in the number of PRDM9 zinc fingers (6 to 12). We sequenced PRDM9 ZF encoding region from 15 individuals (carrying the same ZF number in both copies) and found 43 different ZF domain sequences. Ruminant zinc fingers of PRDM9 were found to be diversifying under positive selection and concerted evolution, specifically at positions involved in defining their DNA-binding specificity, consistent with the reports from other vertebrates such as mice, humans, equids and chimpanzees. ZF-encoding regions of the PRDM7, a paralog of PRDM9 on bovine chromosome 22 and on unknown chromosomes in other studied species were found to contain 84 base repeat units as in PRDM9, but there were multiple disruptive mutations after the first repeat unit. The diversity of the ZFs suggests that PRDM9 may activate recombination hotspots that are largely unique to each ruminant species.</p></div

Gel image of PCR product of ZF domain of PRDM9 and PRDM7 genes in ruminants.

<p>M: 100 bp ladder, bright bands correspond to 500 bp, 1000 bp and 1500 bp. 1–4: upper band- A to D alleles of PRDM9 in cattle, lower band- PRDM7 in cattle. 5–6: upper band- A allele of PRDM9 in yak, lower band- PRDM7 in yak. 7–8: upper band- A and B alleles of PRDM9 in mithun, lower band- PRDM7 in mithun. 9–10: upper band- D allele of PRDM9 in sheep, lower band- PRDM7 in sheep. 11–15: upper band- C to G alleles of PRDM9 in goat, lower band- PRDM7 in goat.</p

Evaluation of the company which successfully passed the reorganization - evaluation of the company MSV Metal Studénka, a.s.

The aim of my diploma thesis is to estimate the market value of the company MSV Metal Studénka, a. s. as of 31st December 2015. The evaluated company produces large variation of steel forgings, pressings and railcar components. The thesis will bring financial analysis of the company and based on strategic analysis and value drivers of the company the financial plan will be drawn up. The valuation of the company will be estimated using income methods of discounted cash flows APV adding market comparison method specifically called similar public company method

dN/dS estimates of variable amino acid sites in ZF domains of PRDM9 gene in ruminants.

<p>Positive selection was estimated in all species combined as well as for individual species separately. Two asterisks indicate P<0.01 and single asterisk indicates P<0.05.</p

Schematic representation of PRDM9 domains and allelic variation in ruminants.

<p>Different ZF repeats in different alleles and species are coded by letters as already shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0156159#pone.0156159.g004" target="_blank">Fig 4</a>. The first finger (shown with green diamond shape) was found to be conserved.</p

Structure Based <i>In Silico</i> Analysis of Quinolone Resistance in Clinical Isolates of <i>Salmonella</i> Typhi from India

<div><p>Enteric fever is a major cause of morbidity in several parts of the Indian subcontinent. The treatment for typhoid fever majorly includes the fluoroquinolone group of antibiotics. Excessive and indiscriminate use of these antibiotics has led to development of acquired resistance in the causative organism <i>Salmonella</i> Typhi. The resistance towards fluoroquinolones is associated with mutations in the target gene of DNA Gyrase. We have estimated the Minimum Inhibitory Concentration (MIC) of commonly used fluoroquinolone representatives from three generations, ciprofloxacin, ofloxacin, levofloxacin and moxifloxacin, for 100 clinical isolates of <i>Salmonella</i> Typhi from patients in the Indian subcontinent. The MICs have been found to be in the range of 0.032 to 8 μg/ml. The gene encoding DNA Gyrase was subsequently sequenced and point mutations were observed in DNA Gyrase in the quinolone resistance determining region comprising Ser83Phe/Tyr and Asp87Tyr/Gly. The binding ability of these four fluoroquinolones in the quinolone binding pocket of wild type as well as mutant DNA Gyrase was computationally analyzed by molecular docking to assess their differential binding behaviour. This study has revealed that mutations in DNA Gyrase alter the characteristics of the binding pocket resulting in the loss of crucial molecular interactions and consequently decrease the binding affinity of fluoroquinolones with the target protein. The present study assists in understanding the underlying molecular and structural mechanism for decreased fluoroquinolone susceptibility in clinical isolates as a consequence of mutations in DNA Gyrase.</p></div

Chemical configuration of quinolones.

<p>2-D chemical structure of second (a) ciprofloxacin and (b) ofloxacin; third (c) levofloxacin and fourth (d) moxifloxacin generation quinolones.</p