A NOVEL METHOD FOR ISOLATION AND TRILINEAGE DIFFERENTIATION OF RAT BONE MARROW DERIVED MESENCHYMAL STEM CELLS

Objective: Goal of this study is to find a simple method for isolation, colony formation and Trilineage differentiation of Mesenchymal stem cells frombone marrow of Wistar Rat. Adherent capacity, morphology, trilineage differentiation and colony formation of bone marrow mesenchymal stem cellswere studied in low glucose, high glucose Dulbecco modified eagle medium with various concentration of fetal bovine serum.Methods: Mesenchymal stem cells were isolated from bone marrow of Wistar rat by Ficoll (sigma) density gradient centrifugation with plasticadherence method. Bone marrow was collected from femur and Tibia of 6-weeks-old Wistar rat; Bone marrow was cultured in Dulbecco's modifiedEagles's medium (DMEM) with low glucose, high glucose supplement (Invitrogen) and 10%, 15% of fetal bovine serum (FBS) at the density of1 × 106, incubated at 37°C in 5% of CO; adherent capacity, colony formation were studied. Differentiation of bone marrow mesenchymal stem cells(BMMSCs) was monitored under suitable differentiation medium. The cells were refeed every 3-4 days and passaged when the cells reached 80-90%confluences.2Result: BMMSCs were adhered in tissue culture flask at 24 hrs, 48 hrs in high glucose and low glucose DMEM with 10% of FBS, respectively. Colonyformation was faster in high glucose and low glucose DMEM with 15% of FBS compared to high glucose and low glucose DMEM with10% FBS.Morphological changes were observed in BMMSCs high glucose, low glucose DMEM with 10% FBS, but no changes were found in the differentiationof BMMSCs in high glucose, low glucose DMEM. Differentiation of BMMSCs was nourishing in third passage cells.Conclusion: High glucose DMEM with 10% FBS is a good supplement for adherence of cells whereas high glucose, low glucose DMEM with 15% FBScan be utilized for rapid Colony formation. Third passage BMMSCs is fruitful for differentiation of BMMSCs.Keywords: Wistar rat, Bone marrow, Mesenchymal stem cells, Adherent capacity, Colony formation, Differentiation, Osteoblast, Chondrocytes,Adipocytes

Seaweed extract as a biostimulant for legume crop, green gram

The aim of this research is to investigate the effect of seaweed extracts obtained from the marine green algae, Ulva lactuca Linnaeus, Caulerpa scalpelliformis (R. Brown ex Turner) C. Agardh, brown algae Sargassum plagiophyllum C. Agardh, Turbinaria conoides (J. Agardh) Kutzing, Padina tetrastromatica Hauck, Dictyota dichotama (Hudson) J. V. Lamouroux on the stimulate germination, growth parameters of the Vigna radiata. The present study reveals the seeds germination, fresh weight and dry weight of shoots and roots. These results suggested that seaweed extracts stronger induce seed germination and growth parameters

Formation of polycrystalline SnS layers by a two-step process

Thin films of SnS have been produced by a novel two-stage process. This involved the deposition of thin films of Sn onto glass substrates using d.c. magnetron sputtering followed by conversion of the metallic layers into the compound by annealing in the presence of elemental sulfur. All the layers synthesised were found to be polycrystalline, the grain size and crystallinity of the layers increasing with increasing annealing temperature. The precursor layers sulfurised at temperatures 350°C, were found to be non-stoichiometric and X-ray diffraction data indicated the presence of a range of binary phases other than SnS. The best SnS layers were synthesised for annealing temperatures between 300 and 350°C. These layers were found to be stoichiomentric with a strong {111} preferred orientation. The stoichiometric SnS layers had resistivities of 1.5×102 ?cm and Arrhenius plots of the resistivity gave an activation energy of 0.65 eV. The optical energy band gap of the layers was 1.35 eV. These p-type layers could find application as absorber layers in thin film solar cells

Hydrogenolysis of sorbitol over Ni, Pt and Ru supported on SBA-15

Hydrogenolysis of sorbitol (15% aqueous solution) has been carried out in a batch reactor over Ni (6 wt%), Pt (1 wt%) and Ru (1 wt%) supported on SBA-15 and carbon coated SBA-15 (SBA-15(C)). For comparison, the three metals have also been supported on activated carbon (AC). The catalysts are characterized by XRD, N2 and H2 adsorption measurements. Addition of Ca(OH)2 to the reaction mixture increases conversion and selectivity for the dihydroxy compounds, 1,2-propanediol (PD) and ethylene glycol (EG). Based on yield of dihydric alcohols (PD+EG), the performance of the catalysts at 220 °C and 60 bar in the presence of Ca(OH)2 is in the order: Ru-AC ~ Ru-SBA-15(C) > Ru-SBA-15 ~ Ni-SBA-15, the yields being 40, 39, 31 and 29 wt%, respectively

Sodium-coupled Monocarboxylate Transporters in Normal Tissues and in Cancer

SLC5A8 and SLC5A12 are sodium-coupled monocarboxylate transporters (SMCTs), the former being a high-affinity type and the latter a low-affinity type. Both transport a variety of monocarboxylates in a Na+-coupled manner. They are expressed in the gastrointestinal tract, kidney, thyroid, brain, and retina. SLC5A8 is localized to the apical membrane of epithelial cells lining the intestinal tract and proximal tubule. In the brain and retina, its expression is restricted to neurons and the retinal pigment epithelium. The physiologic functions of SLC5A8 include absorption of short-chain fatty acids in the colon and small intestine, reabsorption of lactate and pyruvate in the kidney, and cellular uptake of lactate and ketone bodies in neurons. It also transports the B-complex vitamin nicotinate. SLC5A12 is also localized to the apical membrane of epithelial cells lining the intestinal tract and proximal tubule. In the brain and retina, its expression is restricted to astrocytes and Müller cells. SLC5A8 also functions as a tumor suppressor; its expression is silenced in tumors of colon, thyroid, stomach, kidney, and brain. The tumor-suppressive function is related to its ability to mediate concentrative uptake of butyrate, propionate, and pyruvate, all of which are inhibitors of histone deacetylases. SLC5A8 can also transport a variety of pharmacologically relevant monocarboxylates, including salicylates, benzoate, and γ-hydroxybutyrate. Non-steroidal anti-inflammatory drugs such as ibuprofen, ketoprofen, and fenoprofen, also interact with SLC5A8. These drugs are not transportable substrates for SLC5A8, but instead function as blockers of the transporter. Relatively less is known on the role of SLC5A12 in drug transport

Influence of Grain Size on the Band-gap of Annealed SnS Thin Films

The manuscript reports the variation in optical band-gap of vacuum annealed SnS thin films. The samples were characterized by using X-Ray Diffraction, UV-visible Spectroscopy and Raman Analysis. Results show that while annealing does not effect the nano-crystalline sample's lattice structure or unit cell size it does control the grain size. The band-gap (Eg) decreases with increase in grain size. Eg values were found to be very high (1.8-2.5 eV) for samples studied

Deferiprone: Pan-selective Histone Lysine Demethylase Inhibition Activity and Structure Activity Relationship Study

Deferiprone (DFP) is a hydroxypyridinone-derived iron chelator currently in clinical use for iron chelation therapy. DFP has also been known to elicit antiproliferative activities, yet the mechanism of this effect has remained elusive. We herein report that DFP chelates the Fe 2+ ion at the active sites of selected iron-dependent histone lysine demethylases (KDMs), resulting in pan inhibition of a subfamily of KDMs. Specifically, DFP inhibits the demethylase activities of six KDMs - 2A, 2B, 5C, 6A, 7A and 7B - with low micromolar IC 50 s while considerably less active or inactive against eleven KDMs - 1A, 3A, 3B, 4A-E, 5A, 5B and 6B. The KDM that is most sensitive to DFP, KDM6A, has an IC 50 that is between 7- and 70-fold lower than the iron binding equivalence concentrations at which DFP inhibits ribonucleotide reductase (RNR) activities and/or reduces the labile intracellular zinc ion pool. In breast cancer cell lines, DFP potently inhibits the demethylation of H3K4me3 and H3K27me3, two chromatin posttranslational marks that are subject to removal by several KDM subfamilies which are inhibited by DFP in cell-free assay. These data strongly suggest that DFP derives its anti-proliferative activity largely from the inhibition of a sub-set of KDMs. The docked poses adopted by DFP at the KDM active sites enabled identification of new DFP-based KDM inhibitors which are more cytotoxic to cancer cell lines. We also found that a cohort of these agents inhibited HP1-mediated gene silencing and one lead compound potently inhibited breast tumor growth in murine xenograft models. Overall, this study identified a new chemical scaffold capable of inhibiting KDM enzymes, globally changing histone modification profiles, and with specific anti-tumor activities

Molecular targets for the protodynamic action of cis-urocanic acid in human bladder carcinoma cells

<p>Abstract</p> <p>Background</p> <p>cis-urocanic acid (cis-UCA) is an endogenous amino acid metabolite capable of transporting protons from the mildly acidic extracellular medium into the cell cytosol. The resulting intracellular acidification suppresses many cellular activities. The current study was aimed at characterizing the molecular mechanisms underlying cis-UCA-mediated cytotoxicity in cultured cancer cells.</p> <p>Methods</p> <p>5367 bladder carcinoma cells were left untreated or treated with cis-UCA. Cell death was assessed by measuring caspase-3 activity, mitochondrial membrane polarization, formation and release of cytoplasmic histone-associated DNA fragments, and cellular permeabilization. Cell viability and metabolic activity were monitored by colorimetric assays. Nuclear labelling was used to quantify the effects of cis-UCA on cell cycle. The activity of the ERK and JNK signalling pathways was studied by immunoblotting with specific antibodies. Phosphatase activity in cis-UCA-treated cells was determined by assay kits measuring absorbance resulting from the dephosphorylation of an artificial substrate. All statistical analyses were performed using the two-way Student's t-test (p < 0.05).</p> <p>Results</p> <p>Here we report that treatment of the 5637 human bladder carcinoma cells with 2% cis-UCA induces both apoptotic and necrotic cell death. In addition, metabolic activity of the 5637 cells is rapidly impaired, and the cells arrest in cell cycle in response to cis-UCA. Importantly, we show that cis-UCA promotes the ERK and JNK signalling pathways by efficiently inhibiting the activity of serine/threonine and tyrosine phosphatases.</p> <p>Conclusions</p> <p>Our studies elucidate how cis-UCA modulates several cellular processes, thereby inhibiting the proliferation and survival of bladder carcinoma cells. These anti-cancer effects make cis-UCA a potential candidate for the treatment of non-muscle invasive bladder carcinoma.</p

MCT1-mediated transport of a toxic molecule is an effective strategy for targeting glycolytic tumors

There is increasing evidence that oncogenic transformation modifies the metabolic program of cells. A common alteration is the upregulation of glycolysis, and efforts to target glycolytic enzymes for anticancer therapy are under way. Here, we performed a genome-wide haploid genetic screen to identify resistance mechanisms to 3-bromopyruvate (3-BrPA), a drug candidate that inhibits glycolysis in a poorly understood fashion. We identified the SLC16A1 gene product, MCT1, as the main determinant of 3-BrPA sensitivity. MCT1 is necessary and sufficient for 3-BrPA uptake by cancer cells. Additionally, SLC16A1 mRNA levels are the best predictor of 3-BrPA sensitivity and are most elevated in glycolytic cancer cells. Furthermore, forced MCT1 expression in 3-BrPA–resistant cancer cells sensitizes tumor xenografts to 3-BrPA treatment in vivo. Our results identify a potential biomarker for 3-BrPA sensitivity and provide proof of concept that the selectivity of cancer-expressed transporters can be exploited for delivering toxic molecules to tumors.National Institutes of Health (U.S.) (NIH CA103866)Jane Coffin Childs Memorial Fund for Medical Research (Fellowship)National Science Foundation (U.S.) (Fellowship)Howard Hughes Medical Institute (Investigator