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Selected AbstractsGABAA -receptor expression in glioma cells is triggered by contact with neuronal cellsEUROPEAN JOURNAL OF NEUROSCIENCE, Issue 8 2001Michael Synowitz Abstract The expression of functional GABAA -receptors in glioma cells correlates with low malignancy of tumours and cell lines from glioma lack these receptors. Here we show that contact with neurons induces the expression of functional GABAA -receptors. C6 and F98 glioma cell lines were labelled by recombinant expression of enhanced green fluorescent protein injected into rat brain and studied in acute slices after two to three weeks of tumour growth. The cells responded to GABA or the specific agonist, muscimol with a current typical for GABAA -receptors, as studied with the patch-clamp technique. To get insight into the mechanism of GABAA receptor induction, the C6 or F98 cells were co-cultured with neurons, astrocytes, oligodendrocytes and microglia. Glioma cells expressed functional GABAA receptors within 24 h only in cultures where physical contact to neurons occurred. Activation of GABAA -receptors in the co-cultures attenuated glioma cell metabolism while blockade of the receptors increased metabolism. We conclude that with this form of interaction, neurons can influence tumour behaviour in the brain. [source] Intracellular trafficking and release of intact edible mushroom lectin from HT29 human colon cancer cellsFEBS JOURNAL, Issue 7 2000Lu-Gang Yu Our previous studies have shown that the Gal,1,3GalNAc,- (Thomsen,Friedenreich antigen)-binding lectin from the common edible mushroom Agaricus bisporus (ABL) reversibly inhibits cell proliferation, and this effect is a consequence of inhibition of nuclear localization sequence-dependent nuclear protein import after ABL internalization [Yu, L.G., Fernig, D.G., White, M.R.H., Spiller, D.G., Appleton, P., Evans, R.C., Grierson, I., Smith, J.A., Davies, H., Gerasimenko, O.V., Petersen, O.H., Milton, J.D. & Rhodes, J.M. (1999) J. Biol. Chem.274, 4890,4899]. Here, we have investigated further the intracellular trafficking and fate of ABL after internalization in HT29 human colon cancer cells. Internalization of 125I-ABL occurred within 30 min of the lectin being bound to the cell surface. Subcellular fractionation after pulse labelling of the cells with 125I-ABL for 2 h at 4 °C followed by culture of the cells at 37 °C demonstrated a steady increase in radioactivity in a crude nuclear extract. The radioactivity in this extract reached a maximum after 10 h and declined after 20 h. Release of ABL from the cell, after pulse labelling, was assessed using both fluorescein isothiocyanate-labelled ABL and 125I-ABL and was slow, with a t1/2 of 48 h. Most of the 125I-ABL both inside cells and in the medium remained intact, as determined by trichloroacetic acid precipitation and SDS/PAGE, and after 48 h only 22 ± 2% of ABL in the medium and 14 ± 2% inside the cells was degraded. This study suggests that the reversibility of the antiproliferative effect of ABL is associated with its release from cells after internalization. The internalization and subsequent slow release, with little degradation of ABL, reflects the tendency of lectins to resist biodegradation and implies that other endogenous or exogenous lectins may be processed in this way by intestinal epithelial cells. [source] Temperature and Storage Duration Effects on Esterase Activity in Fresh-cut Cantaloupe MelonJOURNAL OF FOOD SCIENCE, Issue 3 2003O. Lamikanra ABSTRACT The effect of storage time and temperature (4 °C or 15 °C) on esterase (ES) activity in fresh-cut cantaloupe melon (Cucumis melo L. var. reticulatus Naud) was determined. Enzymatic activity, after 24 h in storage, was reduced by 40% and 10% in fruit stored at 4 °C and 15 °C, respectively. The ES in cantaloupe melon was determined to be carboxylesterase with 2 isozymes (pI = 6.1 and pI = 9.5) and low thermal stability. Pectin methyl esterase activity in cut fruit also decreased by about 25% at both temperatures after 24 h, but greatly increased after 72 h only in 15 °C stored fruit. ES in cantaloupe melon appears to be regulated by metalloproteases, presumably metallo-exoproteases. [source] In vitro effects of phlorotannins from Ascophyllum nodosum (brown seaweed) on rumen bacterial populations and fermentationJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 13 2009Yuxi Wang Abstract BACKGROUND: Use of brown algae (seaweed) in ruminant diets is increasing, but the effects of its phlorotannins (PT) on rumen microbial ecology have not been determined. Mixed forage (50:25:25 ground barley silage,alfalfa hay,grass hay) was used as substrate in a batch culture ruminal incubation that included PT extracted from Ascophyllum nodosum, with and without polyethylene glycol. Principal ruminal bacteria were quantified using real-time polymerase chain reaction. RESULTS: At 500 µg mL,1, PT reduced growth of Fibrobacter succinogenes by 78%, 83% and 65% at 6, 12 and 24 h (P < 0.001), Ruminococcus albus at 24 h only (P < 0.01) and did not affect R. flavefaciens. Non-cellulolytic bacteria Selenomonas ruminantium, Ruminobacter amylophilus and Prevotella bryantii were increased (P < 0.001) by PT at 12 and 24 h. Effects of PT on fermentation products (gas production, volatile fatty acid profiles and ammonia accumulation) were consistent with alterations in rumen microbial populations. CONCLUSION: The effects of PT on ruminal bacteria were species-dependent, which suggests that diet may mediate PT effects on animal performance. The variation in sensitivity of ruminal bacteria to PT reflects previously reported effects of condensed tannins from terrestrial plants on microbial populations. Copyright © 2009 Crown in the right of Canada. Published by John Wiley & Sons, Ltd [source] Prostaglandin F2, Stimulates Endothelial Nitric Oxide Synthase Depending on the Existence of Bovine Granulosa Cells: Analysis by Co-culture System of Endothelial Cells, Smooth Muscle Cells and Granulosa CellsREPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2008K Shirasuna Contents Prostaglandin F2, (PGF2,) induces luteolysis in the mid but not in the early luteal phase; despite this, both the early and the mid corpus luteum (CL) have PGF2, receptor (FPr). We previously indicated that the luteal blood flow surrounding the CL drastically increases prior to a decrease of progesterone (P) in the cows, suggesting that an acute increase of luteal blood flow may be an early sign of luteolysis in response to PGF2, and that this may be induced by a vasorelaxant nitric oxide (NO). The aim of this study was to investigate the luteal stage-dependent and the site-restricted effect of PGF2, and NO on the mRNA expressions and P secretion. To mimic the local luteal region both of peripheral and central areas of the CL, we utilized co-cultures using bovine aorta endothelial cells (EC), smooth muscle cells (SMC) and luteinizing granulosa cells (GC) or fully-luteinized GC. PGF2, stimulated the expression of endothelial NO synthase (eNOS) mRNA at 0.5 h in mix-cultures of EC and SMC with fully-luteinized GC but not with luteinizing GC. The expression of eNOS mRNA in EC was increased by PGF2, at 1 h only when EC was cultured together with fully-luteinized GC but not with luteinizing GC. In all co-cultures, PGF2, did not affect the mRNA expression of FPr. Treatment of NO donor inhibited P secretion at 0.5 h. In conclusion, the present study suggests that the coexistence of the mature luteal cells (fully-luteinized GC) with EC/SMC may be crucial for acquiring functional NO synthesis induced by PGF2,. [source] | ||||||||||