Changes in the levels of pituitary and steroid hormones in ovine and human ovarian follicular fluid

Changes in the protein and steroid hormones of follicular fluid, aspirated from different follicles of sheep and human ovaries, have been measured and correlated with the size of the follicles. As the fluid contains a number of proteins, steroids have been measured directly and after ether extraction. The follicular fluid concentrations of progesterone and 17 beta-oestradiol measured directly in the fluid increased with the size of the follicles. The levels of free testosterone remained constant in all sizes of follicles, while those of bound hormone showed a 10- to 15-fold increase over the free testosterone concentrations in both the sheep and human follicular fluid. A decrease in the levels of bound testosterone in the fluid of large follicles (LFFL) coincided with the increase in bound 17 beta-oestradiol, suggesting the possible conversion of bound testosterone to oestrogen as the follicle attained maturity. The ratio of follicle-stimulating hormone (FSH) to luteinizing hormone (LH) varied in the fluid obtained from different size follicles, being 1:7 in small (SFFL), 1.3.5 in medium (MFFL) and 1:2.3 in large (LFFL) follicles of sheep ovaries. The LH content of follicular fluid of different size follicles appeared to be the same, with LFFL showing a minor increase over SFFL. In the human, the fluid from medium follicles contained very little LH compared to LFFL. These differences in the pattern of LH levels present in the fluid from different size follicles between human and sheep ovaries presumably reflect species variations in the entry of LH into the follicles.     

Effects of NaCl and iso-osmotic PEG stress on growth,osmolytes accumulation and antioxidant defense in cultured sugarcane cells

In order to discriminate between the ionic and osmotic components of salt stress, sugarcane (Saccharum officinarum L. cv. Co 86032) calli were cultured on media containing NaCl or polyethylene glycol (PEG) 8000 that exerted the same osmotic pressure (−0.7 MPa). PEG stress exposure for 15 days led to significant growth reduction and loss in water content than salt stressed and control tissues. Osmotic adjustment (OA) was observed in callus tissues grown on salt, but was not evident in callus grown on PEG. Oxidative damage to membranes, estimated in terms of accumulation of thiobarbituric acid reactive substances-TBARS and electrolytic leakage was significantly higher in both the stressed calli than the control however, the extent of damage was more in the PEG stressed calli. The stressed callus tissues showed inhibition of ascorbate peroxidase activity, while catalase activity was increased. These results indicate sensitivity of cells to PEG-mediated stress than salt stress and differences in their OA to these two stress conditions. The sensitivity to the osmotic stress indicate that expression of the stress tolerance response requires the coordinated action of different tissues in a plant and hence was not expressed at the cellular level.     

Probing binding mechanism of interleukin-6 and olokizumab: in silico design of potential lead antibodies for autoimmune and inflammatory diseases

Computer-aided antibody engineering has been successful in the design of new biologics for disease diagnosis and therapeutic interventions. Interleukin-6 (IL-6), a well-recognized drug target for various autoimmune and inflammatory diseases such as rheumatoid arthritis, multiple sclerosis, and psoriasis, was investigated in silico to design potential lead antibodies. Here, crystal structure of IL-6 along with monoclonal antibody olokizumab was explored to predict antigen–antibody (Ag???Ab)-interacting residues using DiscoTope, Paratome, and PyMOL. Tyr56, Tyr103 in heavy chain and Gly30, Ile31 in light chain of olokizumab were mutated with residues Ser, Thr, Tyr, Trp, and Phe. A set of 899 mutant macromolecules were designed, and binding affinity of these macromolecules to IL-6 was evaluated through Ag???Ab docking (ZDOCK, ClusPro, and Rosetta server), binding free-energy calculations using Molecular Mechanics/Poisson Boltzman Surface Area (MM/PBSA) method, and interaction energy estimation. In comparison to olokizumab, eight newly designed theoretical antibodies demonstrated better result in all assessments. Therefore, these newly designed macromolecules were proposed as potential lead antibodies to serve as a therapeutics option for IL-6-mediated diseases.     

Linked Cross Sectional Schemes for Estimating Growth of Children

For estimation of growth, the efficiency of linked cross sectional scheme has been compared with pure longitudinal and cross sectional schemes. The relevant estimation theory has been developed and the expressions for the optimum estimators alongwith their variances have been derived. It has been observed that for estimation of growth, the linked cross-sectional scheme has been observed to be less efficient as compared to pure longitudinal scheme but more efficient than pure cross-sectional scheme.     

Biochemical analysis and synergistic suppression of indoxacarb resistance in Plutella xylostella L.

Indoxacarb was treated to Plutella xylostella for 10 generations to develop a resistant strain and biochemical analysis of indoxacarb resistance in different tissues of P. xylostella was carried out. Biochemical analysis found maximum esterase activity in gut homogenates of indoxacarb resistant strains followed by whole body and cuticle homogenates. In gut homogenates of indoxacarb resistant strains, maximum increase in esterases was found as compared to the unselected strain. Acetylcholineesterase activity was higher in head homogenates of the resistant strain than in the unselected strain. Glutathione-S-transferase activity was highest in whole body homogenates. However, maximum increase was found in gut homogenates of indoxacarb resistant strains over the unselected. Induced resistance was suppressed using known synergists. Maximum synergism occurred using diethyl-maleate (DEM), followed by triphenyl phosphate (TPP).     

Tree ring imprints of long-term changes in climate in western Himalaya,India

Tree-ring analyses from semi-arid to arid regions in western Himalaya show immense potential for developing millennia long climate records. Millennium and longer ring-width chronologies of Himalayan pencil juniper (Juniperus polycarpos), Himalayan pencil cedar (Cedrus deodara) and Chilgoza pine (Pinus gerardiana) have been developed from different sites in western Himalaya. Studies conducted so far on various conifer species indicate strong precipitation signatures in ring-width measurement series. The paucity of weather records from stations close to tree-ring sampling sites poses difficulty in calibrating tree-ring data against climate data especially precipitation for its strong spatial variability in mountain regions. However, for the existence of strong coherence in temperature, even in data from distant stations, more robust temperature reconstructions representing regional and hemispheric signatures have been developed. Tree-ring records from the region indicate multi-century warm and cool anomalies consistent with the Medieval Warm Period and Little Ice Age anomalies. Significant relationships noted between mean premonsoon temperature over the western Himalaya and ENSO features endorse utility of climate records from western Himalayan region in understanding long-term climate variability and attribution of anthropogenic impact.     

Assessment of bacterial diversity in agricultural by-product compost by sequencing of cultivated isolates and amplified rDNA restriction analysis

An investigation of bacterial diversity in compost was performed using molecular chronometer in order to reveal its phylogeny. Thirty-three bacterial isolates isolated from compost were analyzed by 16S rRNA gene sequencing which revealed phylogenetic lineage of class Bacilli, γ, β-Proteobacteria, and Actinobacteria. Among these lineages, isolates belonging to class Bacilli consisted of species from genera Staphylococcus, Bacillus, Terribacillus, and Lysinibacillus. From phylum Actinobacteria: Microbacterium barkeri and Kocuria sp. were identified. Other bacterial groups had phylogenetic linkage with genera Comamonas and Acidovorax (class β-Proteobacteria); Serratia, Klebsiella, and Enterobacter (class γ-Proteobacteria). Similar isolates were analyzed through ARDRA. Amplified product of 16S rRNA gene from each isolates was subjected to cleavage by enzymes HpaII, HinfI, and MspI in separate reaction tubes. HpaII generated 2–6 bands ranging from 90–688 bp, HinfI generated 2–5 bands of 71–1,038 bp, and MspI 2–7 bands of 69–793 bp. The restriction patterns from HpaII, HinfI, and MspI were normalized separately and combined by means of pattern recognition software “Diversity Database.” HpaII had highest discrimination index (0.72) than HinfI (0.68) and MspI (0.65), and the combination of all three showed discrimination index (0.69). Numerical analysis of ARDRA patterns demonstrated sufficient phylogenetic information for characterizing bacterial diversity. Phylogenetic relationship obtained among isolates through ARDRA was compared with 16S rRNA gene sequence and ARDRA results showed sufficiently similar 16S rRNA gene sequence analysis, but not an overlapping. It has been observed that ARDRA technique facilitates the identification of bacteria in less than 36 h as compared to traditional 16S rRNA gene sequencing.     

Effect of pH on lipid accumulation by an oleaginous yeast:Rhodotorula glutinis IIP-30

Maximum lipid production (66% w/w dry wt) inRhodotorula glutinis IIP-30 utilizing glucose in a fed-batch fermentation under N-limiting conditions at 30°C, was at pH 4. At pH 3, 5 and 6, the lipid contents were 12%, 48% and 44%, respectively. There was only a small change in the fatty acid profile over the pH range examined, although the ergosterol content decreased by a third as the pH increased.