Aberrant expression of imprinted genes in post-implantation rat embryos

AimImprinted genes are known regulators of embryo growth. Studies from our laboratory have demonstrated that treatment of adult male rats with tamoxifen increased post-implantation loss at around midgestation. Expression of insulin like growth factor 2 (Igf2), a paternally expressed imprinted gene was down-regulated in the resorbing embryos obtained at embryonic day 13. Hypomethylation of Igf2-H19 imprint control region was observed in the resorbing embryo sires and spermatozoa obtained from tamoxifen-treated rats thereby suggesting that errors in imprint acquisition during spermatogenesis can result in embryo loss. The present study aims at studying the expression of other imprinted genes, besides Igf2 in the embryos sired by tamoxifen-treated males.Main methodsGene expression profiles of resorbing versus normal embryos were assessed by microarrays. Real time quantitative RT-PCR for six imprinted genes and four genes involved in cell cycle was done to validate gene expression data. The affected pathways and functions were identified in the resorbing embryos and effect on cell cycle was confirmed by flow cytometry.Key findingsAberrant expression of a number of imprinted genes was observed in the resorbing embryos when compared to the normal embryos at embryonic days 11 and 13. Down-regulation of Notch signaling, Wnt signaling and cell cycle pathway was observed in the resorbing embryos.SignificanceThe study suggests that exposure of male germ cells to tamoxifen during adulthood results in aberrant expression of imprinted genes and down-regulation of development associated pathways in the F₁ progeny thereby causing embryo loss.     

Modulation of mango ripening by chemicals: physiological and biochemical aspects

During ripening, fleshy fruits undergo textural changes that lead to loss of tissue firmness and consequent softening due to cell wall dismantling carried out by different and specifically expressed enzymes. The effect of various chemical treatments on the ripening of mango fruit (Mangifera indica) was investigated at physiological and biochemical level. Based on changes in respiration, firmness, pH, total soluble sugar and a cell wall degrading enzyme pectate lyase (PEL) activity, treatment with 1-methylcyclopropene (1-MCP), silver nitrate (AgNO₃), gibberlic acid (GA₃), sodium metabisulphite (SMS) and ascorbic acid led to delaying of ripening process while those of ethrel and calcium chloride (CaCl₂) enhanced the process. PEL of mango was found to be inhibited by certain metabolites present in dialysed ammonium sulphate enzyme extract as well as EDTA. Mango PEL activity exhibited an absolute requirement for Ca²⁺ and an optimum pH of 8.5.     

Cloning of Salinity Tolerant Gene from a Soil-Borne Bacterium and its Characterization

A salt tolerant gene was isolated and characterized from a highly salt tolerant soil - borne bacterium, Bacillus coagulans. The genomic library was prepared in ZAP expression vector. The amplified library had inserts ranging from 0.5–3.8 kb. The library was screened for isolation of salt tolerant recombinant clones by functional assay using LB agar containing 4% NaCl. Salt tolerant recombinant clone was able to grow much faster than the wild strain on LB agar plates containing 4% NaCl. Size of the gene responsible for salt tolerance was 3.8 kb and had restriction sites for a number of restriction enzymes like HindIII, SmaI, PstI, ClaI, SacI and SpeI. The restriction map of the gene was developed through single and double digestion with the restriction enzymes. The sequence of the insert revealed ORF of 393 amino acids and using BLASTP nr-na database, the insert was identified as ornithine amino transferase, which is known to play an important role in biosynthesis of proline — an important osmolyte involved in salt tolerance of many species of bacteria and plants.     

Cryopreservation and genetic stability assessment of regenerants of the critically endangered medicinal plant Dioscorea deltoidea Wall. ex Griseb. for cryobanking of germplasm

In Vitro Cellular & Developmental Biology - Plant - Dioscorea deltoidea Wall. ex Griseb. is a critically endangered, commercially important crop of high medicinal value. Several accessions of...     

Chemoresistance of cancer floating cells is independent of their ability to form 3D structures: Implications for anticancer drug screening

Three-dimensional (3D) culture systems such as floating spheroids (FSs) and floating tumorspheres (FTs) are widely used as tumor models of chemoresistance. FTs are considered to be enriched in cancer stem-like cells (CS-LCs). In this study, we used cancer cell lines (lung H460, prostate LnCAP, and breast MCF-7) able to form FSs under anchorage-independent conditions and compared with cell lines (prostate PC3 and breast MDA-MB-231) that cannot form FSs under similar conditions. Independent of their ability to form FTs all cell lines growing under anchorage-independent conditions become highly resistant to obatoclax, colchicine, and hydroxyurea. We used anti-E-cadherin antibody (that blocked the formation of FSs) and demonstrated that floating LnCAP cells showed similar chemoresistance regardless of the formation of spheroids. Our results demonstrate that the development of chemoresistance is not because of the formation of a complex 3D structure and/or enrichment of CS-LCs but is likely the result of cell detachment per se and their ability to survive under anchorage-independent conditions. We propose that FSs and FTs could be useful models to study chemoresistance of cancer cells associated with cell detachment (e.g., circulating tumor cells) but they may not be representative of other types of chemoresistance that arise in vivo in attached cells.     

Ecological correlates of Himalayan musk deer Moschus leucogaster

Himalayan musk deer (Moschus leucogaster; hereafter musk deer) are endangered as a result of poaching and habitat loss. The species is nocturnal, crepuscular, and elusive, making direct observation of habitat use and behavior difficult. However, musk deer establish and repeatedly use the same latrines for defecation. To quantify musk deer habitat correlates, we used observational spatial data based on presence–absence of musk deer latrines, as well as a range of fine spatial‐scale ecological covariates. To determine presence–absence of musk deer, we exhaustively searched randomly selected forest trails using a 20‐m belt transect in different study sites within the Neshyang Valley in the Annapurna Conservation Area. In a subsequent way, study sites were classified as habitat or nonhabitat for musk deer. A total of 252 plots, 20 × 20 m, were systematically established every 100 m along 51 transects (each ~0.5 km long) laid out at different elevations to record a range of ecological habitat variables. We used mixed‐effect models and principal component analysis to characterize relationships between deer presence–absence data and habitat variables. We confirmed musk deer use latrines in forests located at higher elevations (3,200–4,200 m) throughout multiple seasons and years. Himalayan birch (Betula utilis) dominated forest, mixed Himalayan fir (Abies spectabilis), and birch forest were preferred over pure Himalayan fir and blue pine (Pinus wallichiana) forest. Greater crown cover and shrub diversity were associated with the presence of musk deer whereas tree height, diameter, and diversity were weakly correlated. Topographical attributes including aspect, elevation, distance to water source, and slope were also discriminated by musk deer. Over‐ and understory forest management can be used to protect forests likely to have musk deer as predicted by the models to ensure long‐term conservation of this rare deer.     

Sulfur stress-induced antioxidative responses in leaves of Triticum aestivum L.

Antioxidative responses were investigated in leaves of wheat (Triticum aestivum L.) grown at varying S levels ranging from deficiency to excess (1, 2, 4, 6 and 8 mM S). Optimum yield was observed in plants supplied with 4 mM S. Wheat responded to S deficiency and excess supply by decreasing growth of root and shoot. Chlorosis in young leaves was observed after 15 days of deficient S supply. The biomass and concentration of photoassimilatory pigments decreased in plants grown at 1, 2, 6 and 8 mM S supply. The concentration of thiobarbituric acid reactive substances (TBARS), cysteine, nonprotein thiol and hydrogen peroxide (H₂O₂) increased in plants grown under S stress. Accumulation of TBARS and H₂O₂ in leaves indicated oxidative damage in S-deficient and S-excess plants. Deficient and excess levels of S showed an increase in the activities of antioxidative enzymes superoxide dismutase (EC 1.15.1.1), catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7), ascorbate peroxidase (EC 1.11.1.11) and glutathione reductase (EC 1.6.4.2).     

Energy Metabolites,Lipid Variables and Lactation Performance of Periparturient Murrah Buffaloes (Bubalus bubalis) Fed on Diet Supplemented with Inorganic Chromium

The aim of this study was to elucidate the effects of chromium (Cr) supplementation as inorganic Cr (CrCl₃?·?6H₂O) on energy balance, lipid peroxidation, and lactation performance in periparturient Murrah buffaloes. Twenty-four multiparous Murrah buffaloes according to lactation, parity, body mass, and expected calving date were divided equally. Experimental buffaloes were randomly assigned to four treatment diets: a control diet and three diets with an inorganic Cr supplementation at 0.5, 1.0, and 1.5 mg of Cr/kg dry matter (DM), respectively from 60 days before expected calving date until 60 days of lactation. Milk productions of buffaloes were recorded every day until 60 days in milk. Blood samples were collected at days ?60, ?45, ?30,?21, ?15, ?7, ?3, 0, 7, 15, 21, 30, 45, and 60 days relative to actual calving for determination of plasma glucose, nonesterified fatty acid (NEFA), thiobarbituric acid reactive substance (TBARS), total cholesterol, total protein, albumin, blood urea nitrogen (BUN), and minerals. Adding inorganic Cr to the diet of Murrah buffaloes increased milk yield. Percentage of fat and total solid yield increased significantly through the experiment in the Cr-supplemented group. At the day of calving, buffaloes showed a decrease in dry matter intake (DMI), plasma glucose, and zinc (Zn) and Cr concentrations. In contrast, plasma NEFA, TBARS, and copper (Cu) levels were found highest at the day of calving among all groups. Cr supplementation increased peripheral blood glucose concentration while decreased level of NEFA and TBARS was recorded in Cr-fed buffaloes. Supplemental Cr had no effect on plasma cholesterol, total protein, albumin, and BUN in periparturient period. Dietary Cr supplementation had positive effect on plasma Cr concentration, but the plasma concentration of Cu, Zn, and iron (Fe) was not affected by different dietary Cr level supplementation. The results suggest that dietary inorganic Cr supplementation improved milk yield by reducing negative energy balance and lipid peroxidation in buffaloes during periparturient period.     

Guidelines for screening,diagnosis and management of hemoglobinopathies

The β-thalassemias and sickle cell disorders are a major health burden in India. Diagnosis and management of these disorders both in adults and in newborns using appropriate approaches and uniform technology are important in different regions of a vast and diverse country as India. In view of a National Thalassemia Control Program to be launched soon, a need was felt for guidelines on whom to screen, cost-effective technologies that are to be used as well as for establishing prenatal diagnosis programs in regional centers. Newborn screening for sickle cell disorders is in its infancy in India and uniform approaches need to be followed. Also, included are guidelines for monitoring and managing patients who are now growing older and need comprehensive care as well as management of complications of the disease.