The glycosphingolipid composition and glycosyltransferase activities of the small intestinal mucosa of testosterone-treated rats

Prior studies have demonstrated that sex hormones can influence the glycosphingolipid composition of different organs, including small intestine. However, to date, the effects of testosterone on glycosphingolipids of rat small intestinal mucosa have not been examined. Experiments were conducted to examine the effect of subcutaneous administration of synthetic testosterone (500 micrograms/100 g body wt.) on the gangliosides and neutral glycosphingolipids of rat small intestinal mucosa. Their results demonstrated that testosterone administrations: (i) increased the ganglioside content including hematoside (GM3); (ii) increased the total content of neutral glycosphingolipids, which was due to the increases in glucosylceramide and globotriaosylceramide; (iii) increased the activities of cytidine 5'-monophosphate-N-acetylneuraminic acid: lactosylceramide sialyltransferase, and UDPgalactose: lactosylceramide galactosyltransferase; (iv) increased the percentage of the long chain base phytosphingosine in hematoside, glucosyl-, and globotriaosylceramide; and (v) significantly altered the fatty acid composition of each of these glycosphingolipids. These results demonstrate that administration of testosterone induces alterations in glycosphingolipid composition and glycosyltransferases activities in rat small intestinal mucosa.     

Drought induces many forms of cysteine proteases not observed during natural senescence

Drought-induced senescence and natural senescence was characterised in the cowpea leaf, with a focus on cysteine proteases. Soluble protein content and ribulose 1,5-bisphosphate carboxylase (Rubisco) content declined as senescence progressed. Endopeptidase activity with Rubisco as a physiological substrate exhibited significant increase at acidic (pH 4.8) than at neutral (pH 7.0) during drought induced senescence and declined during recovery. Natural senescence was associated with a several-fold increase in the endopeptidase activity at both the pHs. Cysteine proteases were analyzed using western blot with polyclonal antibodies raised against papain. Several polypeptides of molecular weights 57, 52, and 43 kDA were recognized by the antibodies, the levels of which showed an increase under water deficit conditions, followed by a decrease during recovery. Three polypeptides of molecular weights 69, 60, and 48 kDa appeared only during the water stress conditions, whereas, during natural senescence, only a single 48 kDa polypeptide with maximum intensity at 9 days after flowering was observed. The results suggests the possibility of distinguishing drought-induced and natural senescence.     

Genetic Analysis of Indian Lentil (Lens culinaris Medikus) Cultivars and Landraces Using RAPD and STMS Markers

Molecular analysis of 29 lentil (Lens culinaris) cultivars and landraces of Indian origin was carried out using twenty RAPD and ten cross-species STMS primers. A total of 97 markers (72 RAPD and 25 STMS) were amplified of which 42.3% were polymorphic. Genetic similarity among the cultivars and landraces was 89.7%. The observed results indicated low level of genetic diversity in the studied material. UPGMA cluster analysis for the combined data of RAPD and STMS revealed two broad clusters — Cluster I with three landraces and Cluster II containing all remaining landraces and cultivars except Precoz. Germplasm line Precoz was found to be the most distinct in individual as well as combined analyses. All cultivars and landraces except K-75 and L4076 could be discriminated from one another using combined data for the two techniques. Germplasm lines Precoz, L830 and cultivars L4147 and JL3 were quite distinct and could be potential germplasm resource.     

Fine needle aspiration cytology of minor salivary gland tumours of the palate

Fine needle aspiration cytology of minor salivary gland tumours of the palate This retrospective study was carried out to review aspirates from minor salivary gland tumours of the palate and to assess the problems encountered in their diagnosis, especially the cytological diagnosis of newer entities such as polymorphous low grade adenocarcinoma (PLGA). Fifty-five cases of palatal salivary gland tumours aspirated over a period of 16 years were reviewed. Histology was available in 26 cases. Pleomorphic adenoma (27 cases) was the most common benign cytodiagnosis. Eleven aspirates were malignant tumours of which eight cases were adenoid cystic carcinoma and three cases were mucoepidermoid carcinoma. Seven cases were diagnosed on fine needle aspiration as suggestive of PLGA. However histological confirmation was available in only one of these cases. Concordance between the initial and revised typings of the tumours was seen in only 28 cases (54%) in the present study. Initially 18 of the 51 tumours (35.3%) could not be typed; and after review, only three could not be typed. Three cases of oncocytoma could be diagnosed on review only. Palatal salivary gland tumours, although relatively uncommon, are difficult to diagnose cytologically. This is more so in cases of newer entities such as PLGA, as their cytological diagnosis is still not well characterized.     

Next Generation Sequencing Analysis Reveals Segmental Patterns of microRNA Expression in Mouse Epididymal Epithelial Cells

The functional maturation of mammalian spermatozoa is accomplished as the cells descend through the highly specialized microenvironment of the epididymis. This dynamic environment is, in turn, created by the combined secretory and absorptive activity of the surrounding epithelium and displays an extraordinary level of regionalization. Although the regulatory network responsible for spatial coordination of epididymal function remains unclear, recent evidence has highlighted a novel role for the RNA interference pathway. Indeed, as noncanonical regulators of gene expression, small noncoding RNAs have emerged as key elements of the circuitry involved in regulating epididymal function and hence sperm maturation. Herein we have employed next generation sequencing technology to profile the genome-wide miRNA signatures of mouse epididymal cells and characterize segmental patterns of expression. An impressive profile of some 370 miRNAs were detected in the mouse epididymis, with a subset of these specifically identified within the epithelial cells that line the tubule (218). A majority of the latter miRNAs (75%) were detected at equivalent levels along the entire length of the mouse epididymis. We did however identify a small cohort of miRNAs that displayed highly regionalized patterns of expression, including miR-204-5p and miR-196b-5p, which were down- and up-regulated by approximately 39- and 45-fold between the caput/caudal regions, respectively. In addition we identified 79 miRNAs (representing ~ 21% of all miRNAs) as displaying conserved expression within all regions of the mouse, rat and human epididymal tissue. These included 8/14 members of let-7 family of miRNAs that have been widely implicated in the control of androgen signaling and the repression of cell proliferation and oncogenic pathways. Overall these data provide novel insights into the sophistication of the miRNA network that regulates the function of the male reproductive tract.     

Whole genome expression array profiling highlights differences in mucosal defense genes in Barrett's esophagus and esophageal adenocarcinoma

Esophageal adenocarcinoma (EAC) has become a major concern in Western countries due to rapid rises in incidence coupled with very poor survival rates. One of the key risk factors for the development of this cancer is the presence of Barrett's esophagus (BE), which is believed to form in response to repeated gastro-esophageal reflux. In this study we performed comparative, genome-wide expression profiling (using Illumina whole-genome Beadarrays) on total RNA extracted from esophageal biopsy tissues from individuals with EAC, BE (in the absence of EAC) and those with normal squamous epithelium. We combined these data with publically accessible raw data from three similar studies to investigate key gene and ontology differences between these three tissue states. The results support the deduction that BE is a tissue with enhanced glycoprotein synthesis machinery (DPP4, ATP2A3, AGR2) designed to provide strong mucosal defenses aimed at resisting gastro-esophageal reflux. EAC exhibits the enhanced extracellular matrix remodeling (collagens, IGFBP7, PLAU) effects expected in an aggressive form of cancer, as well as evidence of reduced expression of genes associated with mucosal (MUC6, CA2, TFF1) and xenobiotic (AKR1C2, AKR1B10) defenses. When our results are compared to previous whole-genome expression profiling studies keratin, mucin, annexin and trefoil factor gene groups are the most frequently represented differentially expressed gene families. Eleven genes identified here are also represented in at least 3 other profiling studies. We used these genes to discriminate between squamous epithelium, BE and EAC within the two largest cohorts using a support vector machine leave one out cross validation (LOOCV) analysis. While this method was satisfactory for discriminating squamous epithelium and BE, it demonstrates the need for more detailed investigations into profiling changes between BE and EAC.     

Photoluminescence and structural analysis of trivalent europium doped MLaAl3O7 (M = Ba,Ca, Mg and Sr) nanophosphors

The solution combustion technique was used to synthesize MLaAl₃O₇ (M = Ba, Ca, Mg, and Sr) nanophosphors‐doped with Eu³⁺ using metal nitrates as precursors. The photoluminescence (PL) emission spectra exhibited three peaks at 587–591, 610–616, and 653–654 corresponding to ⁵D₀→⁷F₁, ⁵D₀→⁷F₂, and ⁵D₀→⁷F₃ transitions, respectively. Upon excitation at 254 nm, these nanophosphors displayed strong red emission with the dominant peak attributed to the ⁵D₀→⁷F₂ transition of Eu³⁺. The materials were further heated at 900 and 1050°C for 2 h to examine the consequence of temperature on crystal lattice and PL emission intensity. X‐ray diffraction (XRD) analysis proved that all the synthesized materials were of a crystalline nature. CaLaAl₃O₇ material has a tetragonal crystal structure with space group P421m. Scherer's equation was used to calculate the crystallite size of synthesized phosphors using XRD data. A Fourier transformation infrared study was used to observe the stretching vibrations of metal–oxygen bonds. Infrared peaks for stretching vibrations corresponding to lanthanum–oxygen and aluminium–oxygen bonds were found at 582 and 777 cm^–1 respectively for CaLaAl₃O₇ phosphor material. Transmission electron microscopy images were used to determine the size of particles (18–37 nm for the as‐prepared materials) and also to analyze the three‐dimensional view of these materials. The experimental data indicate that these materials may be promising red‐emitting nanophosphors for use in white light‐emitting diodes.     

Imaging in vivo neuronal transport in genetic model organisms using microfluidic devices

Microfluidic devices have been developed for imaging behavior and various cellular processes in Caenorhabditis elegans, but not subcellular processes requiring high spatial resolution. In neurons, essential processes such as axonal, dendritic, intraflagellar and other long-distance transport can be studied by acquiring fast time-lapse images of green fluorescent protein (GFP)-tagged moving cargo. We have achieved two important goals in such in vivo studies namely, imaging several transport processes in unanesthetized intact animals and imaging very early developmental stages. We describe a microfluidic device for immobilizing C. elegans and Drosophila larvae that allows imaging without anesthetics or dissection. We observed that for certain neuronal cargoes in C. elegans, anesthetics have significant and sometimes unexpected effects on the flux. Further, imaging the transport of certain cargo in early developmental stages was possible only in the microfluidic device. Using our device we observed an increase in anterograde synaptic vesicle transport during development corresponding with synaptic growth. We also imaged Q neuroblast divisions and mitochondrial transport during early developmental stages of C. elegans and Drosophila, respectively. Our simple microfluidic device offers a useful means to image high-resolution subcellular processes in C. elegans and Drosophila and can be readily adapted to other transparent or translucent organisms.     

Spectroscopic and molecular docking studies on chlorambucil interaction with DNA

Chlorambucil (CMB) is an anticancer drug used for the treatment of variety of cancers. Structural and conformational changes associated with DNA after binding with CMB were explored using spectroscopic techniques to get insight into the mechanism of action of CMB at molecular level. Different molar ratios of CMB-DNA complex were prepared with constant DNA concentration under physiological conditions. FTIR spectroscopy, UV-visible spectroscopy, CD spectroscopy and molecular docking studies were employed to determine the binding site and binding constant of CMB with DNA. The results show CMB binds DNA through nitrogenous bases (thymine, guanine and cytosine). The binding constant was calculated to be 1.3×10(3)M(-1), which suggests weak binding of CMB with DNA double helix. FTIR and CD results show that CMB do not disturb native B-conformation of DNA and it continues to remain in its B conformation even at higher concentrations of CMB. The molecular docking results are in corroboration with our experimental results and provides structural insight into the interaction site.