Influence of four host‐plants on feeding,growth and reproduction of Diacrisia casignetum (Lepidoptera: Arctiidae)

Effects of four host‐plants, sunflower, castor, jute and sesame, on feeding, growth and reproduction of Diacrisia casignetum Kollar (Lepidoptera: Arctiidae) were studied under laboratory conditions (27 ± 0.5°C, 12 h light : 12 h dark, 65 ± 5% RH). Total larval developmental time of D. casignetum was highest on sesame than the other three host‐plants used in this study, but pupal duration was higher on sesame than sunflower but not for other dietary treatments. The longevity of females was generally longer than males. Male and female longevity was higher in sunflower than sesame (P < 0.05), but it did not differ significantly among other treatments. Fecundity was highest in sunflower followed by castor, jute and sesame. The growth and development of D. casignetum were related to nutrient and phenol contents of these four host‐plants. Total carbohydrates and amino acids were present in rich quantities in sunflower when compared to other three host leaves, while nitrogen, protein and lipid contents were comparatively higher in sunflower and castor than jute and sesame. Phenol content was greatest in sesame, and least in castor and sunflower. Higher levels of total carbohydrates, proteins, lipids, nitrogen and amino acids including water content and lower phenol content of sunflower have influenced higher growth rate and fecundity of D. casignetum.     

Adaptability in protein structures: structural dynamics and implications in ligand design

The basic framework of understanding the mechanisms of protein functions is achieved from the knowledge of their structures which can model the molecular recognition. Recent advancement in the structural biology has revealed that in spite of the availability of the structural data, it is nontrivial to predict the mechanism of the molecular recognition which progresses via situation-dependent structural adaptation. The mutual selectivity of protein–protein and protein–ligand interactions often depends on the modulations of conformations empowered by their inherent flexibility, which in turn regulates the function. The mechanism of a protein’s function, which used to be explained by the ideas of ‘lock and key’ has evolved today as the concept of ‘induced fit’ as well as the ‘population shift’ models. It is felt that the ‘dynamics’ is an essential feature to take into account for understanding the mechanism of protein’s function. The design principles of therapeutic molecules suffer from the problems of plasticity of the receptors whose binding conformations are accurately not predictable from the prior knowledge of a template structure. On the other hand, flexibility of the receptors provides the opportunity to improve the binding affinity of a ligand by suitable substitution that will maximize the binding by modulating the receptors surface. In this paper, we discuss with example how the protein’s flexibility is correlated with its functions in various systems, revealing the importance of its understanding and for making applications. We also highlight the methodological challenges to investigate it computationally and to account for the flexible nature of the molecules in drug design.     

An investigation into the altered binding mode of green tea polyphenols with human serum albumin on complexation with copper

Green tea is rich in several polyphenols, such as (?)-epicatechin-3-gallate (ECG), (?)-epigallocatechin (EGC), and (?)-epigallocatechin-3-gallate (EGCG). The biological importance of these polyphenols led us to study the major polyphenol EGCG with human serum albumin (HSA) in an earlier study. In this report, we have compared the binding of ECG, EGC, and EGCG and the Cu(II) complexes of EGCG and ECG with HSA. We observe that the gallate moiety of the polyphenols plays a crucial role in determining the mode of interaction with HSA. The binding constants obtained for the different systems are 5.86?±?0.72?×?10⁴ M^?1 (K _ECG-HSA), 4.22?±?0.15?×?10⁴ M^?1 (K _{ECG-Cu(II)-HSA}), and 9.51?±?0.31?×?10⁴ M^?1 (K _{EGCG-Cu(II)-HSA}) at 293?K. Thermodynamic parameters thus obtained suggest that apart from an initial hydrophobic association, van der Waals interactions and hydrogen bonding are the major interactions which held together the polyphenols and HSA. However, thermodynamic parameters obtained from the interactions of the copper complexes with HSA are indicative of the involvement of the hydrophobic forces. Circular dichroism and the Fourier transform infrared spectroscopic measurements reveal changes in α-helical content of HSA after binding with the ligands. Data obtained by fluorescence spectroscopy, displacement experiments along with the docking studies suggested that the ligands bind to the residues located in site 1 (subdomains IIA), whereas EGC, that lacks the gallate moiety, binds to the other hydrophobic site 2 (subdomain IIIA) of the protein.     

Assessment of Anxiety Level and Mental Health Status in Spouses and Children of Day-Working and Shift-Working Men

The present study was aimed at assessing the effects of the unusual schedules of shift-working men on anxiety and mental health of their spouses and children. The anxiety and mental health of 396 subjects that include 133 spouses and 263 children of day-working and shift-working men were monitored by employing specific inventory. Anxiety scales (trait, state and free-floating) and mental health inventory were administrated to each subject. Results indicate that the average levels of free-floating anxiety for the spouses and children of shift-working men were found to be significantly higher in comparison with that of their counterparts sampled in the family of day workers. This was also true when data were segregated with respect of gender and compared. Except in female children of shift workers all other groups of children had statistically significantly higher level of state anxiety as compared with that of their respective mothers. The results of this study further reveal that the spouses of shift-working men exhibited lower mental health rating as compared with the spouses of day-working men. Furthermore, a significant negative relationship was noticed between anxiety and mental health in all cases excepting in spouses of shift-working men for state anxiety. Although the findings of the present study could be attributed to the irregular routine of shift workers, it appears mandatory to carryout further intensive studies to draw convincing conclusions.     

Olfactory responses of Epilachna dodecastigma (Coleoptera: Coccinellidae) to long-chain fatty acids from Momordica charantia leaves

Extraction, thin-layer chromatography and gas chromatography–mass spectrophotometry analyses revealed the presence of 12, 13, and 12 fatty acids in young, mature, and senescent leaves of Momordica charantia L., representing 87.30, 95.25, and 83.11 % of the total fatty acids, respectively. The proportion of saturated fatty acids was highest in senescent leaves (78.60 %) followed by young leaves (69.42 %) and mature leaves (48.92 %), with the balance accounted for by unsaturated fatty acids. Palmitic acid was the predominant saturated fatty acid in the three types of leaves, whereas alpha-linolenic acid was the predominant unsaturated fatty acid. The fatty acids from young, mature, and senescent leaves followed by the application of a synthetic mixture of fatty acids that was comparable to the natural fatty acids found in the three types of leaves, elicited the attraction of the female insect Epilachna dodecastigma (Coleoptera: Coccinellidae) at 50–200, 50–200, and 100–200 μg/ml concentrations, respectively, in a Y-shaped glass tube olfactometer bioassay. Individual synthetic fatty acids were also evaluated by the olfactometer bioassay at concentrations comparable to the proportions detected in the three types of leaves. Individual synthetic palmitic acid, stearic acid, oleic acid, linoleic acid, and alpha-linolenic acid at 58.24, 13.96, 29.40, 30.31, and 29.76 μg, respectively, attracted the insect. A synthetic blend of 79.13, 10.57, 29.40, 30.31, and 36.33 μg of palmitic, stearic, oleic, linoleic, and alpha-linolenic acids, respectively, which is the proportion present in a 200 μg/ml concentration of fatty acids of mature leaves, or of 116.49, 13.96, and 29.76 μg of palmitic, stearic and alpha-linolenic acids, respectively, which is the proportion present in a 200 μg/ml concentration of natural fatty acids of young leaves, served as attractants for E. dodecastigma.     

Neem Leaf Glycoprotein Activates CD8+ T Cells to Promote Therapeutic Anti-Tumor Immunity Inhibiting the Growth of Mouse Sarcoma

In spite of sufficient data on Neem Leaf Glycoprotein (NLGP) as a prophylactic vaccine, little knowledge currently exists to support the use of NLGP as a therapeutic vaccine. Treatment of mice bearing established sarcomas with NLGP (25 µg/mice/week subcutaneously for 4 weeks) resulted in tumor regression or dormancy (Tumor free/Regressor, 13/24 (NLGP), 4/24 (PBS)). Evaluation of CD8⁺ T cell status in blood, spleen, TDLN, VDLN and tumor revealed increase in cellular number. Elevated expression of CD69, CD44 and Ki67 on CD8⁺ T cells revealed their state of activation and proliferation by NLGP. Depletion of CD8⁺ T cells in mice at the time of NLGP treatment resulted in partial termination of tumor regression. An expansion of CXCR3⁺ and CCR5⁺ T cells was observed in the TDLN and tumor, along with their corresponding ligands. NLGP treatment enhances type 1 polarized T-bet expressing T cells with downregulation of GATA3. Treg cell population was almost unchanged. However, T∶Treg ratios significantly increased with NLGP. Enhanced secretion/expression of IFNγ was noted after NLGP therapy. In vitro culture of T cells with IL-2 and sarcoma antigen resulted in significant enhancement in cytotoxic efficacy. Consistently higher expression of CD107a was also observed in CD8⁺ T cells from tumors. Reinoculation of sarcoma cells in tumor regressed NLGP-treated mice maintained tumor free status in majority. This is correlated with the increment of CD44^hiCD62L^hi central memory T cells. Collectively, these findings support a paradigm in which NLGP dynamically orchestrates the activation, expansion, and recruitment of CD8⁺ T cells into established tumors to operate significant tumor cell lysis.     

Fluoroquinolone Resistance Mechanisms of Shigella flexneri Isolated in Bangladesh

Objective

To investigate the prevalence and mechanisms of fluoroquinolone resistance in Shigella species isolated in Bangladesh and to compare with similar strains isolated in China.

Methods

A total of 3789 Shigella isolates collected from Clinical Microbiology Laboratory of icddr,b, during 2004–2010 were analyzed for antibiotic susceptibility. Analysis of plasmids, plasmid-mediated quinolone-resistance genes, PFGE, and sequencing of genes of the quinolone-resistance-determining regions (QRDR) were conducted in representative strains isolated in Bangladesh and compared with strains isolated in Zhengding, China. In addition, the role of efflux-pump was studied by using the efflux-pump inhibitor carbonyl cyanide-m-chlorophenylhydrazone (CCCP).

Results

Resistance to ciprofloxacin in Shigella species increased from 0% in 2004 to 44% in 2010 and S. flexneri was the predominant species. Of Shigella spp, ciprofloxacin resistant (Cip^R) strains were mostly found among S. flexneri (8.3%), followed by S. sonnei (1.5%). Within S. flexneri (n = 2181), 14.5% were resistance to ciprofloxacin of which serotype 2a was predominant (96%). MIC of ciprofloxacin, norfloxacin, and ofloxacin were 6–32 mg/L, 8–32 mg/L, and 8–24 mg/L, respectively in S. flexneri 2a isolates. Sequencing of QRDR genes of resistant isolates showed double mutations in gyrA gene (Ser⁸³Leu, Asp⁸⁷Asn/Gly) and single mutation in parC gene (Ser⁸⁰Ile). A difference in amino acid substitution at position 87 was found between strains isolated in Bangladesh (Asp⁸⁷Asn) and China (Asp⁸⁷Gly) except for one. A novel mutation at position 211 (His→Tyr) in gyrA gene was detected only in the Bangladeshi strains. Susceptibility to ciprofloxacin was increased by the presence of CCCP indicating the involvement of energy dependent active efflux pumps. A single PFGE type was found in isolates from Bangladesh and China suggesting their genetic relatedness.

Conclusions

Emergence of fluoroquinolone resistance in Shigella undermines a major challenge in current treatment strategies which needs to be followed up by using empirical therapeutic strategies.     

Evaluation of the effect of cagPAI genes of Helicobacter pylori on AGS epithelial cell morphology and IL-8 secretion

Helicobacter pylori cagPAI genes play an important role in pathogenesis, however little is known about their functions in isolates from Turkish patients. We aimed to evaluate the intactness and the effect of the cagPAI genes (cagT, cagM, cagE, cagA) and cagA EPIYA motifs on the AGS morphological changes and IL-8 induction. Of 53 patients 38 were found infected with H. pylori. PCR amplification of the cagPAI genes showed 42.1 % intact, 39.5 % partially deleted and 18.4 % with complete deletions. Isolates from gastritis, duodenal and gastric ulcer patients with intact and partially deleted cagPAI genes induced higher IL-8 secretion than those with complete deletions. Isolates from gastritis patients had higher deletion frequencies of the cagT and cagM genes than the other two genes. Infection of AGS cells with isolates that possess intact cagPAI and EPIYA-ABC resulted in the formation of the hummingbird phenotype. The cagA positive isolates induced higher IL-8 secretion than cagA negative isolates. Isolates from DU patients with more than one EPIYA-C motif induced higher concentrations of IL-8 than those with EPIYA-ABC. In conclusion, the intactness of the cagPAI in our isolates from different patients was not conserved. An intact cagPAI was found to play an important role in the pathogenesis of DU but not GU or gastritis. The cagA gene, but not other cagPAI genes, was associated with the induction of IL-8 and the morphological changes of the AGS cells. An increase in the number of EPIYA-C motifs had noticeable effect on the formation of the hummingbird phenotype.     

Seasonal alteration in prevalence and intensity of naturally occurring gastrointestinal helminth infection in goats of New Alluvial zone of West Bengal,India

Parasitic gastroenteritis is one of the major limiting factors for optimum productivity in small ruminants. The present study was carried out to determine the seasonal alteration in prevalence of gastrointestinal helminths (GIH) and intensity of gastrointestinal nematode (GIN) infection in goats of New Alluvial zone (NAZ) of West Bengal. A total of 4320 faecal samples of goats from six different places of NAZ were examined by the standard coprological techniques for the presence of gastrointestinal parasites during January 2014 to December 2015. The meteorological data of the study area were also collected from the Regional Meteorological Station, Kolkata. The prevalence of GIH was recorded as 70.52% and Strongyle (65.18%) group of nematodes was found significantly (p < 0.05) higher than other helminths. Highest (p < 0.05) prevalence (85.11%) as well as intensity of GIN infection (EPG = 640) was observed in monsoon and lowest (p < 0.05) prevalence (60.55%) and intensity (EPG = 340) was recorded in summer. Haemonchus contortus was found to be the predominant GIN species. The observations of the present study both in terms of parasite epidemiology and meteorological data should be utilised for planning strategic control measures against GI helminths in goats of NAZ of West Bengal.     

Molecular characterization of novel Bradyrhizobium strains nodulating Eriosema chinense and Flemingia vestita,important unexplored native legumes of the sub-Himalayan region (Meghalaya) of India

Root nodule bacterial strains were isolated from the little-studied legumes Eriosema chinense and Flemingia vestita (both in tribe Phaseoleae, Papilionoideae) growing in acidic soil of the sub-Himalayan region of the Indian state of Meghalaya (ME), and were identified as novel strains of Bradyrhizobium on the basis of their 16S rRNA sequences. Seven isolates selected on the basis of phenotypic characters and assessment of ARDRA and RAPD patterns were subjected to multilocus sequence analysis (MLSA) using four protein-coding housekeeping genes (glnII, recA, dnaK and gyrB). On the basis of 16S rRNA phylogeny as well as a concatenated MLSA five strains clustered in a single separate clade and two strains formed novel lineages within the genus Bradyrhizobium. The phylogenies of the symbiotic genes (nodA and nifH) were in agreement with the core gene phylogenies. It appears that genetically diverse Bradyrhizobium strains are the principal microsymbionts of these two important native legumes. The novel genotypes of Bradyrhizobium strains isolated in the present study efficiently nodulate the Phaseoloid crop species Glycine max, Vigna radiata and Vigna umbellata. These strains are genetically different from strains of Bradyrhizobium isolated earlier from a different agro-climatic region of India suggesting that the acidic nature of the soil, high precipitation and other local environmental conditions are responsible for the evolution of these newly-described Bradyrhizobium strains. In global terms, the sub-Himalayan region of India is geographically and climatically distinct and the Bradyrhizobium strains nodulating its legumes appear to be novel and potentially unique to the region.