Androgen deprivation by flutamide modulates uPAR, MMP-9 expressions, lipid profile, and oxidative stress: amelioration by daidzein

The growth and development of prostate gland is governed by testosterone. Testosterone helps in maintaining the adipose tissue stores of the body. It is well documented that with advancing age there has been a gradual decline in testosterone levels. Our aim was to study the protective role of daidzein on flutamide-induced androgen deprivation on matrix degrading genes, lipid profile and oxidative stress in Wistar rats. Sub-chronic (60 days) flutamide (30 mg/kg b.wt) administration resulted in marked increase in expressions of matrix degrading genes [matrix metalloproteases 9 and urokinase plasminogen activation receptor]. Additionally, it increased the levels of low density lipoproteins, total cholesterol, triglycerides, and lowered the levels of high density lipoproteins and endogenous antioxidant levels. Oral administration of daidzein (20 and 60 mg/kg b.wt) restituted the levels to normal. Daidzein administration resulted in amelioration of the prostate atrophy, degeneracy and invasiveness induced by flutamide. Our findings suggest that the daidzein may be given as dietary supplement to patients who are on androgen deprivation therapy, to minimize the adverse effects related to it and also retarding susceptibility of patients to cardiovascular diseases.     

Bio-efficacy of some leaf extracts on the inhibition of egg hatching and mortality of Meloidogyne incognita

Nematicidal activities of extracts from plants were assayed against Meloidogyne incognita in vitro. Leaves of six different plants were collected in and around Aligarh Muslim University Campus. Aqueous extracts of six plants were screened for egg hatchability and nematicidal activity against second stage juveniles of M. incognita in the plant pathology and nematology laboratory, AMU Aligarh. The nematode egg and juveniles were exposed 12, 24 and 48 h in (S, S/2, S/10, S/100) concentrations of plant extracts. The plant extracts of leaves of six plants species viz. Jatropha pandurifolia, Polyalthia longifolia, Wedelia chinensis, Nerium indicum, Duranta repens and Cassia fistula exhibited highly promising mortality of 99.00–72.00% after 48 h of exposure. Aqueous extracts of leaves of J. pandurifolia, P. longifolia, W. chinensis were recorded to be highly effective for inhibition of egg hatching and increasing juvenile mortality of M. incognita. There was a gradual decrease in egg hatching and increase in mortality rate of juveniles of M. incognita with increase in the concentration of leaf extract and exposure time.     

The stress-induced SCP/HLIP family of small light-harvesting-like proteins (ScpABCDE) protects Photosystem II from photoinhibitory damages in the cyanobacterium <Emphasis Type="Italic">Synechocystis</Emphasis> sp. PCC 6803

Small CAB-like proteins (SCPs) are single-helix light-harvesting-like proteins found in all organisms performing oxygenic photosynthesis. We investigated the effect of growth in moderate salt stress on these stress-induced proteins in the cyanobacterium Synechocystis sp. PCC 6803 depleted of Photosystem I (PSI), which expresses SCPs constitutively, and compared these cells with a PSI-less/ScpABCDE^? mutant. SCPs, by stabilizing chlorophyll-binding proteins and Photosystem II (PSII) assembly, protect PSII from photoinhibitory damages, and in their absence electrons accumulate and will lead to ROS formation. The presence of 0.2 M NaCl in the growth medium increased the respiratory activity and other PSII electron sinks in the PSI-less/ScpABCDE^? strain. We postulate that this salt-induced effect consumes the excess of PSII-generated electrons, reduces the pressure of the electron transport chain, and thereby prevents ¹O₂ production.     

Deep-diving of Atlantic salmon (<Emphasis Type="Italic">Salmo salar</Emphasis>) during their marine feeding migrations

Data from seven data storage tags recovered from Atlantic salmon marked as smolts were analyzed for depth movements and patterns of deep diving during the marine migration. The salmon mostly stayed at the surface and showed diurnal activity especially from autumn until spring. During the first months at sea the salmon stayed at shallower depths (<100 m). The salmon took short deep dives (>100 m), that were rare or absent during the first summer at sea but increased in frequency and duration especially in late winter. The maximum depth of the dives varied from 419 to 1187 m. Most of dives were short, (<5 h) but could last up to 33 h. The duration of dives increased in late winter until spring and the overall depth and maximum depth per dive increased exponentially over time. The initiation of the dives was more common in evenings and at night, suggesting nocturnal diving. We hypothesized that deep diving is related to feeding of salmon as mesopelagic fish can be important food for salmon during winter.     

Effect of sperm exposure time on in vitro fertilization and embryo development of bovine oocytes matured in vitro

This study was designed to investigate the effect of sperm exposure time on the fertilization rate and subsequent developmental capacity of bovine oocytes matured in vitro. Cumulus oocyte complexes (COCs) obtained from 2 to 6 mm follicles were matured for 24 h in TCM-199 supplemented with fetal bovine serum (FBS) and hormones (FSH, LH and estradiol 17-beta). In vitro fertilization (IVF) was performed by incubating 15 to 20 matured oocytes with 1 x 10(6) percoll separated frozen-thawed spermatozoa in 1 ml of IVF-TL medium for either 4, 8, 12, 16, 20, 24 or 28 h. Following sperm exposure for different periods of times, the presumptive zygotes were co-cultured with Buffalo Rat Liver cells (BRLC) monolayers in CZB medium without glucose, a simple semi-defined medium developed for mouse embryo culture, for 3 d post-insemination and then in M199/FBS (TCM-199-HEPES supplemented with 20% heat-treated FBS and 1 mM sodium pyruvate) for 5 d. The fertilization rates differed significantly among the 7 treatment groups, with higher frequencies obtained by co-incubation of gametes for 20, 24 or 28 h (67 to 76%) than for 4, 8 and 12 h (26 to 54.5%), with 16 h (57%) being intermediate. However, the length of sperm exposure time did not significantly affect subsequent embryo development, although an increasing trend was noted from 4 h to 20 h. The number of fertilized oocytes at 3 d post-insemination cleaving to 2- to 4-cell vs 8-cell stage was not different among treatment groups. Development of 8-cell embryos to morulae and blastocysts did not differ among the treatment groups. These data suggest that the optimum duration of sperm-oocyte incubation is 24 h, and periods shorter than 16 h may result in a reduced fertilization rate.     

Genome-wide association studies of net form of net blotch resistance at seedling and adult plant stages in spring barley collection

Net form of net blotch (NFNB) of barley (Hordeum vulgare L.), caused by Pyrenophora teres f. teres (Ptt) Drechsler (anamorph: Drechslera teres [Sacc.] Shoem.), is considered one of the major constraints of successful barley production in major barley growing regions of the world. Resistance to NFNB was evaluated in a barley collection of 336 genotypes (AM-2014), at seedling stage using isolates LGDPtt.19 and TD10 in the USA, and adult stage in seven hotspot environments in Morocco. The AM-2014 panel was genotyped with 9K SNP markers and genome-wide association studies (GWAS) were carried out using mixed linear model (MLM: Q?+?K) accounting for population structure (Q) and kinship (K) as covariates. Significant (P?<?0.001) marker trait associations were corrected for false discovery rate (FDR) at the q?<?0.05. Four genotypes showed an average infection response (IRs ≤ 2) to both isolates, LGDPttt.19 and TD10, at the seedling stage, and 30 genotypes showed resistance in all environments in the field while three genotypes exhibited the highest resistance at both stages. The GWAS of NFNB identified 31 distinct QTLs on all seven barley chromosomes, of which 8 with resistance at seedling stage, 21 were associated with resistance at the adult stage, and two QTLs, QRptt.2H-132.15 and QPtt.6H-54-55, conferred resistance at both stages. Of 31 resistance QTLs reported in this study, 10 QTLs coincided with previously mapped QTL while 21 are novel, thereby validating the GWAS approach used in this study. The resistance sources identified in AM-2014 and QTL mapped in this study are valuable resources for marker-assisted breeding for NFNB resistance in the future.     

Molecular Elucidation of Two Novel Seed Specific Flavonoid Glycosyltransferases in Soybean

Flavonoids are specialized plant secondary metabolites that are mainly present as glycoconjugates and function as attractants to pollinators and symbionts, UV protectants, allelochemicals, and have antimicrobial and antiherbivore activity for plant health. Because of the heterogeneity of UDP-glycosyltransferases (UGTs) for glycosylation in plants, their function in flavonoid glycosylation remains largely unknown in soybean and other legumes, particularly that of the UGT92 genes. Here, we identified 152 putative UGT92 genes across 48 plant species and elucidated their mode of duplication, expansion/deletion pattern, alignment, phylogenetic analysis, and genome-wide distribution. Two novel UGT-encoding genes Glyma14g04790 (UGT92G1) and Glyma15g03670 (UGT92G2) were isolated from soybean and their heterologous expression was optimized in Escherichia. coli. Both genes exhibited catalytic activity toward quercetin, kaempferol, and myricetin, with UDPglucose as the sugar donor and were characterized as flavanol-specific UGTs. High expression of both UGTs was observed in adaxial and abaxial parenchyma, suspensor cells, and adaxial and abaxial epidermis cells during seed development, suggesting that they are seed-specific flavanol glycosyltransferases in soybean. Co-expression analysis of UGT92 genes with their first and second neighborhood genes provided a basis for their network elucidation in soybean. We provide valuable information on the role of UGT92 in seed development via the glycosylation of multiple flavanols and the potential metabolic engineering of flavonoid compounds in both plants and E. coli.     

Inhibition of tumour and non-tumour cell proliferation by pygidial gland secretions of four ground beetle species (Coleoptera: Carabidae)

Inhibition of the proliferation of human tumour cells and porcine non-tumour cells by the pygidial gland secretion released by adults of four ground beetle species was observed in this study. The sulphorhodamine B (SRB) assay was applied to establish the percentages of inhibition of the net growth of four human tumour cell lines and porcine liver primary non-tumour cells. The secretions of all tested ground beetle species were shown to have an antiproliferative effect on the tested cell lines. Special emphasis is put on the secretion of Abax parallelepipedus, which showed the highest antitumour potential and weakest inhibition of non-tumour cell proliferation. The antitumour and antiproliferative potential of the pygidial gland secretions of ground beetles is here demonstrated for the first time. It is suggested that certain organic acids are responsible for the action. Further investigation needs to be conducted in order to better understand the mechanisms governing the observed cytotoxic and antitumour activity.     

Cloning,antibody production,expression and cellular localization of universal stress protein gene (<Emphasis Type="Italic">USP1</Emphasis>-GFP) in transgenic cotton

This study was aimed to clone the universal stress protein (GUSP1) gene isolated from Gossypium arboreum in E. coli expression vector pET30(a) and to raise the specific antibody in rabbit to devise a system that could be used for localization and expression of this gene under drought stress. The amplification of GUSP1 transgene revealed a fragment of 500 bp via PCR in genomic DNA of transgenic cotton plants and expression was confirmed through ELISA and Western blot by using the GUSP1 specific polyclonal antibodies. ELISA showed the expression of GUSP1 protein in roots, stem and leaves of transgenic plants at seedling, vegetative and mature plant developmental stages. Total protein isolated from drought stressed transgenic plants revealed a fragment of 47 kDa (GUSP1-GFP fusion protein) in Western blot which confirmed the expression of transgene. Confocal microscopy detected the GFP fluorescence as localization of GUSP1 in the midrib, guard cells of stomata, trichome and globular trichome of intact leaf of transgenic plants. The co-localization was observed within cytoplasm, palisade, spongy mesophyll, guard cells of stomata, vascular bundle, trichome and globular trichome of transgenic plants by using the GUSP1 specific primary antibodies and Alexa fluor conjugated secondary antibodies. This study of GUSP1 gene will advance the mechanism of abiotic stress tolerance in plants.     

Highly selective anthraquinone-chalcone hybrids as potential antileukemia agents

A series of 23 novel anthraquinone-chalcone hybrids containing amide function was synthesized and structurally characterized. Sixteen compounds exerted strong cytotoxic activities against K562, Jurkat and HL-60 leukemia cell lines and significantly lower cytotoxic effects against normal MRC-5 cells, indicating very high selectivity in their anticancer action. The compounds 6g, 6u and 6v activate apoptosis in K562 cells through the extrinsic and intrinsic apoptotic pathway. The compound 6e triggered apoptosis in K562 cells only through the extrinsic apoptotic pathway. Treatment of K562 cells with each of these four compounds caused decrease in the expression levels of MMP2, MMP9, and VEGF, suggesting their anti-invasive, antimetastatic and antiangiogenic properties. The compounds 6g and 6v downregulated expression levels of miR-155 in K562 cells, while compounds 6e and 6u upregulated miR-155 levels in treated cells, in comparison with control cells. The structure-based 3-D QSAR models for 6f, 6e, 6i and 6l describe pro-apoptotic activity against caspase-3.