Home About us Contact
|
Home About us Contact | ||
|
|
||
Complete Inhibition (complete + inhibition)
Selected AbstractsIn Vivo RANK Signaling Blockade Using the Receptor Activator of NF-,B:Fc Effectively Prevents and Ameliorates Wear Debris-Induced Osteolysis via Osteoclast Depletion Without Inhibiting OsteogenesisJOURNAL OF BONE AND MINERAL RESEARCH, Issue 2 2002Lisa M. Childs Abstract Prosthesis failure due to wear debris-induced osteolysis remains a major clinical problem and the greatest limitation for total joint arthroplasty. Based on our knowledge of osteoclast involvement in this process and the requirements of receptor activator of NF-,B (RANK) signaling in osteoclastogenesis and bone resorption, we investigated the efficacy of RANK blockade in preventing and ameliorating titanium (Ti)-induced osteolysis in a mouse calvaria model. Compared with placebo controls we found that all doses of RANK:Fc above 1 mg/kg intraperitoneally (ip) per 48 h significantly inhibited osteoclastogenesis and bone resorption in response to Ti implanted locally. Complete inhibition occurred at 10 mg/kg ip per 48 h, yielding results that were statistically equivalent to data obtained with Ti-treated RANK,/, mice. We also evaluated the effects of a single injection of RANK:Fc on day 5 on established osteolysis and found that Ti-treated were still depleted for multinucleated tartrate-resistant acid phosphatase-positive (TRAP+) cells 16 days later. More importantly, this osteoclast depletion did not affect bone formation because the bone lost from the osteolysis on day 5 was restored by day 21. An assessment of the quantity and quality of the newly formed bone in these calvariae by calcein labeling and infrared (IR) microscopy, respectively, showed no significant negative effect of RANK:Fc treatment. These studies indicate that osteoclast depletion via RANK blockade is an effective method to prevent and reverse wear debris-induced osteolysis without jeopardizing osteogenesis. [source] Effects of tea polyphenols on the activities of soybean trypsin inhibitors and trypsinJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 2 2004Huihua Huang Abstract Tea polyphenols (TPs) and other materials were extracted from Chinese green tea, and their effects on trypsin inhibitors and trypsin were analysed. TPs were found to have a deactivation effect on both Kunitz trypsin inhibitor (KTI) and Bowman,Birk trypsin inhibitor (BBTI). KTI was more easily deactivated than BBTI by complexing with TPs. The deactivation effect of TPs on KTI and BBTI reached a maximum at a TP/KTI ratio of 25 and a TP/BBTI ratio of 16. However, the deactivation effect of TPs on KTI and BBTI was reduced dramatically when KTI and BBTI were already complexed with trypsin. TPs were also found to inhibit trypsin. The inhibitory activity of TPs, KTI and BBTI on trypsin was found to decrease in the order BBTI > gtTI > gtPs. Complete inhibition of trypsin by TPs could not be achieved. When the TP concentration was increased to about 17 µg ml,1, the residual activity of trypsin was maintained at 400 TU mg,1, equivalent to 32% of the initial trypsin activity. In TP inhibition the KM value for trypsin remained unchanged at 5.88 × 10,4 mol l,1 and Vmax decreased when benzoyl- DL -arginine- p -nitroanilide (BAPNA) was used as substrate. The pattern of trypsin inhibition by TPs is non-competitive. Copyright © 2004 Society of Chemical Industry [source] Complete inhibition of fibrinolysis by sustained carboxypeptidase B activity: the role and requirement of plasmin inhibitorsJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 6 2007J. B. WALKER Summary.,Background:,The antifibrinolytic effect of activated thrombin-activatable fibrinolysis inhibitor (TAFIa) and carboxypeptidase B (CPB) displays threshold behavior. When CPB was used to simulate conditions mimicking continuous TAFIa activity, it affected the lysis of plasma clots differently to clots formed from a minimal fibrinolytic system comprising fibrinogen, plasminogen and ,2 -antiplasmin. Whereas CPB saturably prolonged clot lysis in the purified system, the effect of CPB did not appear saturable in plasma clots. Methods:,To rationalize this difference, we investigated the effects of ,2 -antiplasmin, ,2 -macroglobulin, antithrombin and aprotinin on CPB-mediated antifibrinolysis. Results:,CPB alone prolonged fibrinolysis in a saturable manner and the efficacy of CPB increased with decreasing tissue-type plasminogen activator (t-PA) concentration. The inhibitors by themselves did not halt fibrinolysis and the potency of each inhibitor in the absence of CPB mirrored their solution-phase plasmin inhibitory potentials: ,2 -antiplasmin , aprotinin >> ,2 -macroglobulin >> antithrombin. With both CPB and inhibitor present, a synergistic effect was observed. The antifibrinolytic sensitivity to CPB was related to the plasmin inhibitory potential of the inhibitor. Conclusions:,Fibrinolysis could be completely inhibited by ,2 -antiplasmin, ,2 -macroglobulin and antithrombin, but not aprotinin, in the presence of CPB, and occurred only when the irreversible inhibitor or pool of inhibitors were in excess of plasminogen. Western blot analysis indicated that the CPB-mediated shutdown of fibrinolysis was a result of plasminogen consumption prior to clot lysis. The CPB concentration required for fibrinolytic shutdown was dependent on t-PA concentration and the inhibitory potential of the irreversible inhibitor pool. [source] A Human Anti-CD40 Monoclonal Antibody, 4D11, for Kidney Transplantation in Cynomolgus Monkeys: Induction and Maintenance TherapyAMERICAN JOURNAL OF TRANSPLANTATION, Issue 8 2009T. Aoyagi Blockade of CD40,CD154 signaling pathway is an attractive strategy to induce potent immunosuppression and tolerance in organ transplantation. Due to its strong immunosuppressive effect shown in nonhuman primate experiments, anti-CD154 monoclonal antibodies (mAbs) have been tried in clinical settings, but it was interrupted by unexpected thromboembolic complications. Thus, inhibition of the counter molecule, CD40, has remained an alternative approach. In the previous preliminary study, we have shown that 4D11, a novel fully human anti-CD40 mAb, has a fairly potent immunosuppressive effect on kidney allograft in nonhuman primates. In this study, we aimed to confirm the efficacy and untoward events of the 2-week induction and 180-day maintenance 4D11 treatments. In both, 4D11 significantly suppressed T-cell-mediated alloimmune responses and prolonged allograft survival. Addition of weekly 4D11 administration after the induction treatment further enhanced graft survival. Complete inhibition of both donor-specific Ab and anti-4D11 Ab productions was obtained only with higher-dose maintenance therapy. No serious side effect including thromboembolic complications was noted except for a transient reduction of hematocrit in one animal, and decrease of peripheral B-cell counts in all. These results indicate that the 4D11 appears to be a promising candidate for immunosuppression in clinical organ transplantation. [source] Coxiella burnetii inhabits a cholesterol-rich vacuole and influences cellular cholesterol metabolismCELLULAR MICROBIOLOGY, Issue 3 2006Dale Howe Summary Coxiella burnetii directs the synthesis of a large parasitophorous vacuole (PV) required for replication. While some lysosomal characteristics of the PV have been described, the origin and composition of the PV membrane is largely undefined. Cholesterol is an essential component of mammalian cell membranes where it plays important regulatory and structural roles. Here we investigated the role of host cholesterol in biogenesis and maintenance of the C. burnetii PV in Vero cells. The C. burnetii PV membrane stained with filipin and was positive for the lipid raft protein flotillin-1, suggesting PV membranes are enriched in cholesterol and contain lipid raft microdomains. C. burnetii infection increased host cell cholesterol content by 1.75-fold with a coincident upregulation of host genes involved in cholesterol metabolism. Treatment with U18666A, lovastatin, or 25-hydroxycholesterol, pharmacological agents that inhibit cholesterol uptake and/or biosynthesis, altered PV morphology and partially inhibited C. burnetii replication. Complete inhibition of C. burnetii PV development and replication was observed when infected cells were treated with imipramine or ketoconazole, inhibitors of cholesterol uptake and biosynthesis respectively. We conclude that C. burnetii infection perturbs host cell cholesterol metabolism and that free access to host cholesterol stores is required for optimal C. burnetii replication. [source] ,-Amyloid immunization approaches for Alzheimer's diseaseDRUG DEVELOPMENT RESEARCH, Issue 2 2002Bruno P. Imbimbo Abstract Alzheimer's disease (AD) represents the third leading cause of death in the U.S. and the leading cause of dementia in the elderly population. Until recently, there was little hope of efficiently combating this devastating disease. The deposition of ,-amyloid (A,) is the major pathological hallmark of AD brains. Genetic, biochemical, and pharmacological evidence support the hypothesis that A, plays a key role in the development of the disease. Thus, in the last 5 years a number of pharmacological strategies have been developed to interfere with the A, cascade. The most revolutionary of these approaches was proposed in 1999 by scientists at Elan Pharmaceuticals, which immunized against A, transgenic mice with spontaneously developing A, pathology. The immunization was achieved by subcutaneous injections of a preaggregated form of the synthetic human 42-amino acid A, emulsified with Freund's adjuvant, an immune stimulant. The vaccination caused a near complete inhibition of A, plaque formation in younger animals and a marked reduction of the A, burden in older animals. The effects on A, plaques were accompanied by a reduction of A,-associated astrogliosis and neuritic dystrophy. These results were later confirmed by other groups with similar vaccination protocols, which also demonstrated that the A, immunization of transgenic animals normalize or reduce the cognitive impairment associated with A, pathology. Interestingly, effective removal of brain A, plaques was also obtained by peripherally administering A, antibodies. The mechanism with which the vaccine increases A, clearance is not fully understood. Centrally, the vaccine appears to activate A, phagocytosis by microglial monocytes. Peripherally, serum A, antibodies bind and sequester A,, thus altering its equilibrium between CNS and plasma. The dramatic results obtained in animal models of AD raised unprecedented hopes for both a preventive and a curative intervention for this devastating disorder. A vaccine preparation for human use (AN-1792) composed of preaggregated human A,42 peptide and a highly purified saponin derivative (QS-21) was developed by Elan Pharmaceuticals and Wyeth Ayerst and tested in AD patients. Unfortunately, a Phase IIa study aimed at evaluating the safety and immunological activity of AN-1792 in 360 AD patients was discontinued because 15 subjects receiving the vaccine developed serious signs of CNS inflammation. Both central activation of cytotoxic T cells and autoimmune reactions were proposed as potential mechanisms of toxicity. Other therapeutic A, vaccination strategies are being pursued, including immuno-conjugates and monoclonal antibodies. The future of these and other A, immunization approaches depend on a clear understanding of the mechanism of A, clearance and additional insight into the role of inflammation in the AD brain. Drug Dev. Res. 56:150,162, 2002. © 2002 Wiley-Liss, Inc. [source] Glucose inhibits the formation of gas vesicles in Haloferax volcanii transformantsENVIRONMENTAL MICROBIOLOGY, Issue 1 2008Torsten Hechler Summary The effect of glucose on the formation of gas vesicles was investigated in Haloferax mediterranei and Hfx.volcanii transformants containing the mc- gvp gene cluster of Hfx. mediterranei (mc-vac transformants). Increasing amounts of glucose in the medium resulted in a successive decrease in the amount of gas vesicles in both species, with a complete inhibition of their formation at glucose concentrations of > 70 mM in mc-vac transformants, and 100 mM in Hfx. mediterranei. Maltose and sucrose imposed a similar inhibitory effect, whereas xylose, arabinose, lactose, pyruvate and 2-deoxy-glucose had no influence on the gas vesicle formation in mc-vac transformants. The activities of the two mc-vac promoters were strongly reduced in mc-vac transformants grown in the presence of > 50 mM glucose. The gas vesicle overproducing ,D transformant (lacking the repressing protein GvpD) also showed a glucose-induced lack of gas vesicles, indicating that GvpD is not involved in the repression. The addition of glucose was useful to block gas vesicle formation at a certain stage during growth, and vice versa, gas vesicle synthesis could be induced when a glucose-grown culture was shifted to medium lacking glucose. Both procedures will enable the investigation of defined stages during gas vesicle formation. [source] Effect of byproducts from the ozonation of pyrene: Biphenyl-2,2,,6,6,-tetracarbaldehyde and biphenyl-2,2,,6,6,-tetracarboxylic acid on gap junction intercellular communication and neutrophil functionENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 3 2005Stephanie L. Luster-Teasley Abstract In this study, biphenyl-2,2,,6,6,-tetracarbaldehyde, an initial by product formed from the ozonation of pyrene, and biphenyl-2,2,,6,6,-tetracarboxylic acid, a subsequent pyrene ozonation byproduct, were evaluated using two toxicology assays to compare the toxicity of ozonation byproducts with that of the parent compound. The first assay measured the potential for the compounds to block gap junctional intercellular communication (GJIC) using the scrape loading/dye transfer technique in normal WB-344 rat liver epithelial cells. The second assay evaluated the ability of the compounds to affect neutrophil function by measuring the production of superoxide in a human cell line (HL-60). Pyrene significantly blocked intercellular communication (f= 0.2,0.5) at 40 ,M and complete inhibition of communication (f < 0.2) occurred at 50 ,M. Gap junctional intercellular communication in cells exposed to biphenyl-2,2,,6,6,-tetracarbaldehyde reached f < 0.5 at a concentration of 15 ,M. At concentrations greater than 20 ,M, biphenyl-2,2,,6,6,-tetracarbaldehyde was cytotoxic and the inhibition of GJIC was caused by cell death. Biphenyl-2,2,,6,6,-tetracarboxylic acid was neither cytotoxic nor inhibitory to GJIC at the concentrations tested (10,500 ,M). Exposure to biphenyl-2,2,,6,6,-tetracarbaldehyde resulted in a concentration-dependent decrease in phorbol 12-myristate 13-acetate,stimulated O12 production. Neither exposure to pyrene nor biphenyl-2,2,,6,6,-tetracarboxylic acid caused a significant toxic effect on neutrophil function. [source] Toxicity of lead in aqueous medium to Desulfovibrio desulfuricans G20ENVIRONMENTAL TOXICOLOGY & CHEMISTRY, Issue 2 2003Rajesh K. Sani Abstract The toxicity of Pb(II) to sulfate-reducing bacteria (SRB) was studied using Desulfovibrio desulfuricans G20 in a medium specifically designed to assess metal toxicity. The effects of Pb(II) toxicity were observed in terms of longer lag times, lower specific growth rates, and in some cases no measurable growth. With an increase in medium pH from 6 to 8, Pb(II) toxicity decreased. At all pH values, in the presence of Pb(II) concentrations ranging from 3 to 15 ,M, specific growth rates decreased and lag times increased. The minimum inhibiting concentration (MIC) of Pb(II) causing a complete inhibition in growth at pH 6 was 10 ,M, as compared to 15 ,M at pH 7.2 and 8. These MIC values are 40 times lower than previously reported for SRB. Results also show that with increases in initial cell protein concentration (inoculum size), soluble Pb(II) removal rates increased and the degree to which Pb(II) caused increased lag times was reduced. In the presence of Pb(II), in all cases in which D. desulfuricans grew (even after a 312-h lag time), the final cell protein concentration was equivalent to that of the Pb-free control. Live/dead staining, based on membrane integrity, indicated that while Pb(II) inhibited growth, Pb(II) did not cause a loss of D. desulfuricans membrane integrity. [source] Phospholipase,C, negatively regulates Rac/Cdc42 activation in antigen-stimulated mast cellsEUROPEAN JOURNAL OF IMMUNOLOGY, Issue 1 2007Mirvat El-Sibai M.D. Abstract The Rho GTPases Rac and Cdc42 play a central role in the regulation of secretory and cytoskeletal responses in antigen-stimulated mast cells. In this study, we examine the kinetics and mechanism of Rac and Cdc42 activation in the rat basophilic leukemia RBL-2H3 cells. The activation kinetics of both Rac and Cdc42 show a biphasic profile, consisting of an early transient peak at 1,min and a late sustained activation phase at 20,40,min. The inhibition of phospholipase,C (PLC), causes a twofold increase in Rac and Cdc42 activation that coincides with a dramatic production of atypical filopodia-like structures. Inhibition of protein kinase,C using bisindolylmaleimide mimics the effect of PLC, inhibition on Rac activation, but not on Cdc42 activation. In contrast, depletion of intracellular calcium leads to a complete inhibition of the early activation peak of both Rac and Cdc42, without significant effects on the late sustained activation. These data suggest that PLC, is involved in a negative feedback loop that leads to the inhibition of Rac and Cdc42. They also suggest that the presence of intracellular calcium is a prerequisite for both Rac and Cdc42 activation. [source] Content and biosynthesis of polyamines in salt and osmotically stressed cells of Synechocystis sp.FEMS MICROBIOLOGY LETTERS, Issue 1 2003PCC 680 Abstract The effects of various NaCl and sorbitol concentrations in the growth medium on polyamine content and on two enzymes of the polyamine biosynthesis pathway, arginine decarboxylase (ADC) and S -adenosyl methionine decarboxylase (SAMDC), were investigated in the unicellular cyanobacterium Synechocystis sp. PCC 6803. Synechocystis cells showed no difference in growth rate when the concentration of NaCl was raised up to 550 mM. The growth rate decreased at 300 mM sorbitol, and complete inhibition of growth occurred at concentrations of ,700 mM sorbitol. Salt stress induced a moderate increase in the total cellular polyamine content, spermine in particular. Osmotic stress caused an apparent increase in the total cellular polyamine content with a marked increase of spermidine induced by 700 mM sorbitol. Importantly, a low level of spermine, which so far has never been detected in cyanobacteria, could be found in Synechocystis sp. PCC 6803. ADC, a key enzyme for putrescine synthesis, was unaffected by salt stress but showed a six-fold increase in enzyme activity upon osmotic stress imposed by 700 mM sorbitol. SAMDC, another important enzyme for spermidine and spermine synthesis, responded to salt and osmotic stresses similarly to the pattern observed for ADC. An analysis by reverse transcription-polymerase chain reaction revealed an increase of ADC mRNA level in cells under salt and osmotic stresses. Most importantly, the increase of ADC mRNA was attributed to its slower turnover rate under both stress conditions. Interestingly, the samdc gene(s) of Synechocystis appear to be unique since comparisons with known gene sequences from other organisms resulted in no homologous sequences identified in the Synechocystis genome. [source] Inhibition of accelerated tumor growth by blocking the recruitment of mobilized endothelial progenitor cells after chemotherapyINTERNATIONAL JOURNAL OF CANCER, Issue 7 2009Junichi Murakami Abstract It has been suggested that immature progenitor cells mobilize from bone marrow into the peripheral blood in response to the chemotherapy-induced myelosuppression. We investigated how the mobilization of immature progenitor cells affects tumor growth after chemotherapy. We found significantly increased numbers of CD34+/Flk-1+ endothelial progenitor cells in the peripheral blood of mice 1 week after the administration of 100 mg/kg cyclophosphamide vs. a saline injection (0.39 ± 0.09% vs. 0.20 ± 0.10%, respectively; p < 0.05). Tumor growth in the mice given chemotherapy was almost 1.3-fold faster than that in the mice given saline (268 ± 66 mg vs. 210 ± 3 5 mg, respectively; p < 0.05). Histological examination of tumor tissue revealed significantly higher microvessel density and more Ki67-positive cells, but significantly fewer apoptotic cells, in the mice given chemotherapy than in those given saline (p < 0.05). Furthermore, we detected significantly more bone marrow-derived cells, some of which stained positively for CD34 and were localized in the vessels, in tumor tissue from the mice given chemotherapy than in that from the mice given saline. However, the transient disruption of the SDF-1/CXCR4 axis by the antibody neutralization of CXCR4, which occurred over 1 week, blocked the recruitment of bone marrow-derived cells into the tumor tissue, and resulted in complete inhibition of accelerated tumor growth after chemotherapy. Our results show that chemotherapy induced the mobilization of endothelial progenitor cells and accelerated tumor growth, but that transient disruption of the SDF-1/CXCR4 axis could prevent accelerated tumor growth by blocking the recruitment of mobilized endothelial progenitor cells after chemotherapy. © 2008 Wiley-Liss, Inc. [source] A recombinant humanized anti-insulin-like growth factor receptor type I antibody (h7C10) enhances the antitumor activity of vinorelbine and anti-epidermal growth factor receptor therapy against human cancer xenograftsINTERNATIONAL JOURNAL OF CANCER, Issue 2 2005Liliane Goetsch Abstract Interaction of insulin-like growth factor receptor I (IGF-IR) with its ligands has been reported to induce cell proliferation, transformation and blockade of cell apoptotic functions. IGF-IR is overexpressed on numerous tumor cell types and its blockade could be of importance for anti-cancer therapy. We have generated a humanized anti-IGF-IR antibody h7C10 that blocks in vitro IGF-I and IGF-II-induced cell proliferation of MCF-7 breast cancer cells. Analysis of the IGF-I transduction cascade demonstrated that the humanized anti-IGF-IR antibody and its murine parental form block IGF-I-induced tyrosine phosphorylation, both its ,-chain and IRS-1 tyrosine phosphorylation. This presumably leads to cell cycle arrest and, consequently, growth inhibition. Treatment of nude mice bearing either human breast cancer cells (MCF-7) or non small lung cancer cells (A549) with h7C10, or its murine parental form 7C10, inhibited significantly tumor growth. An almost complete inhibition of A549 tumor growth was observed when mice were treated with the anti-IGF-IR antibody combined with either a chemotherapeutic agent, Vinorelbine or an anti-epidermal growth factor receptor (EGFR) antibody, 225. Combined therapy prolonged significantly the life span of mice in an orthotopic in vivo model of A549; the combination of the anti-IGF-IR antibody with an anti-EGFR antibody was superior to the Vinorelbine combination. The present results indicate that the humanized anti-IGF-IR antibody h7C10 has a great potential for cancer therapy when combined with either a chemotherapeutic agent or an antibody that targets other growth factor receptors, such as the epidermal growth factor receptor. [source] Potential of peanut skin phenolic extract as antioxidative and antibacterial agent in cooked and raw ground beefINTERNATIONAL JOURNAL OF FOOD SCIENCE & TECHNOLOGY, Issue 7 2010Jianmei Yu Summary This study investigated the potential of peanut skin extract (PSE) as inhibitor of lipid oxidation in cooked and raw ground beef (GB) and as antimicrobial agent in raw GB. Results show that addition of PSE to raw GB before cooking significantly inhibited the formation of peroxides and TBARS in cooked GB during the refrigerated storage. PSE at concentration ,0.06% was as effective as BHA/BHT at 0.02% in inhibiting lipid oxidation. PSE also inhibited the oxidation of meat pigments thereby preserving the fresh redness of treated meat when used at 0.02,0.10%. Microplate assay showed complete inhibition of test bacteria (Bacillus subtilis, Salmonella typhimurium, Staphylococcus aureus, Streptococcus faecalis and Escherichia coli) in the presence of PSE at 0.4% or higher. However, the antimicrobial effect of PSE in GB was less potent. Hence, PSE can primarily serve the dual purposes of preserving the colour of raw GB and preventing lipid oxidation in cooked products. [source] Efficacy of natamycin for control of growth and ochratoxin A production by Aspergillus carbonarius strains under different environmental conditionsJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2007Á. Medina Abstract Aims:, To examine the efficacy of natamycin produced by Streptomyces natalensis against strains of Aspergillus carbonarius growth and ochratoxin A (OTA) production under different environmental factors on a grape juice-based medium. Methods and Results:, Detailed studies in the range 0,20 ng ml,1 for control of growth and ochratoxin production by strains of A. carbonarius at 0·98, 0·96 and 0·94 water availabilities (aw) and 15,25°C on a fresh red grape extract medium were examined. Inhibition of growth was depending on temperature and aw level. At 15°C, 5,10 ng ml,1 natamycin was effective in reducing growth almost completely. However, at 20,25°C and all the three aw levels, growth was only slightly inhibited by 5,10 ng ml,1 natamycin. There were strain differences with regard to inhibition of OTA production. At 15°C and 0·98 aw, 10 ng ml,1 was required to inhibit production by >90%. However, at 0·96 and 0·94 aw, almost complete inhibition occurred. At 20°C, OTA production was only significantly inhibited by 10 ng ml,1 natamycin at 0·94 aw. At 0·96 and 0·98 aw, some inhibition occurred with 5,10 ng ml,1, but greater concentrations would be required for effective inhibition. At 25°C, 5 ng ml,1 was effective at all aw levels. However, at 15°C and 25°C and a wide range of aw levels, natamycin effectively controlled OTA production. Conclusions:, Natamycin appears to be a very effective for controlling growth and OTA production by strains of A. carbonarius over a range of aw and temperature conditions on grape-based media. Significance and Impact of the Study:, This is the first detailed study to demonstrate the impact of natamycin against A. carbonarius. This study suggests that use of natamycin at 50,100 ng ml,1 can give complete inhibition of growth of A. carbonarius and OTA production over a range of environmental conditions. Natamycin could be an important component of a system to prevent OTA contamination of wine as well during the drying and production of vine fruits. [source] Effect of temperature and pH on the toxicity of aluminium towards two new, soil born species of Arthrobacter sp.JOURNAL OF BASIC MICROBIOLOGY, Issue 2 2004Paul Illmer Dr. Two coryneform bacteria Arthrobacter sp. PI/1,95 (Arth1) and Arthrobacter sp. PI/3-95 (Arth3) were isolated from forest soil characterized and investigated with respect to their reaction towards aluminium (Al). Sigmoid functions were used to describe dose-response relationships between Al concentration and microbial growth and to calculate EC-values. EC100 , indicating a complete inhibition of microbial growth , varied between 185 ,M Al for Arth1 and 11 mM Al for Arth3. A pure pH effect seems probable in connection with the sensitive Arth1 but not with the tolerant species Arth3. Temperature was shown to distinctly increase the toxic effects of Al towards Arth3 whereas only a moderate modification in Al-toxicity was observed with Arth1. (© 2004 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Inhibition of human cytochrome p450 1b1 further clarifies its role in the activation of dibenzo[a,l]pyrene in cells in cultureJOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 3 2007Brinda Mahadevan Abstract Metabolic activation and DNA adduct formation of the carcinogenic aromatic hydrocarbon dibenzo{a,l}pyrene (DBP) was investigated in human mammary carcinoma MCF-7 cells and human cytochrome P450 (CYP) 1B1-expressing Chinese hamster V79 cells in culture. It has been shown that DBP is metabolically activated to DNA-binding diol epoxides both in vitro and in vivo. To further establish the role of human CYP1B1 in the activation of DBP, both cell lines were cotreated with DBP and a selective chemical inhibitor of CYP1B1, 2,4,3, ,5,-tetramethoxy-stilbene (TMS). Results from DBP,DNA adduct analyses revealed the complete inhibition of DNA binding when cells were cotreated with DBP and TMS in comparison to DBP alone. Inactivation of CYP1B1 by TMS was also demonstrated through a decrease in the 7-ethoxyresorufin O -deethylase (EROD) activity in microsomes isolated from these cells. Emodin, 3-methyl-1,6,8-trihydroxyanthraquinone, an active ingredient of an herb, has been recently shown of being able to induce CYP1 gene expression. Examination of human CYP1B1 induction and EROD activity confirmed an increase in protein levels upon cotreatment with emodin and DBP. Despite increases in protein levels and enzyme activity, there was no significant change in DBP,DNA binding levels at very low substrate concentrations (17 nM). The data obtained in this study emphasize the central role of CYP1B1 in the activation of DBP in human cells in culture. © 2007 Wiley Periodicals, Inc. J Biochem Mol Toxicol 21:101,109, 2007; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20168 [source] Role of annexin A6 isoforms in catecholamine secretion by PC12 cells: Distinct influence on calcium responseJOURNAL OF CELLULAR BIOCHEMISTRY, Issue 1 2010Paulina Podszywalow-Bartnicka Abstract Noradrenaline and adrenaline are secreted by adrenal medulla chromaffin cells via exocytosis. Exocytosis of catecholamines occurs after cell stimulation with various endogenous activators such as nicotine or after depolarization of the plasma membrane and is regulated by calcium ions. Cytosolic [Ca2+] increases in response to cell excitation and triggers a signal-initiated secretion. Annexins are known to participate in the regulation of membrane dynamics and are also considered to be involved in vesicular trafficking. Some experimental evidence suggests that annexins may participate in Ca2+ -regulated catecholamine secretion. In this report the effect of annexin A6 (AnxA6) isoforms 1 and 2 on catecholamine secretion has been described. Overexpression of AnxA6 isoforms and AnxA6 knock-down in PC12 cells were accompanied by almost complete inhibition or a 20% enhancement of dopamine secretion, respectively. AnxA6-1 and AnxA6-2 overexpression reduced ,[Ca2+]c upon depolarization by 32% and 58%, respectively, while AnxA6 knock-down increased ,[Ca2+]c by 44%. The mechanism of AnxA6 action on Ca2+ signalling is not well understood. Experimental evidence suggests that two AnxA6 isoforms interact with different targets engaged in regulation of calcium homeostasis in PC12 cells. J. Cell. Biochem. 111: 168,178, 2010. © 2010 Wiley-Liss, Inc. [source] Reduction of intracellular pH inhibits constitutive expression of Cyclooxygenase-2 in human colon cancer cellsJOURNAL OF CELLULAR PHYSIOLOGY, Issue 2 2004Daniela Pirkebner Cyclooxygenase-2 (COX-2) over-expression is critically involved in tumor formation. Intracellular pH (pHi) has been shown to be alkaline in cancer cells, and to be an important trigger for cell proliferation. This study therefore analyzed the relationship between pHi and COX-2 expression. HRT-18 and Caco-2 cells cultured in medium with bicarbonate maintained a pHi of ,7.6, which is higher than that of non-neoplastic cells. Cells grown in bicarbonate-free medium with a pH at 6.8 showed a reduction in pHi to approximately 7.0. Importantly, reduction of pHi resulted in a complete inhibition of COX-2 mRNA and protein expression. When cells were grown in bicarbonate-supplemented medium at pH 6.8, pHi maintained at ,7.6 and COX-2 expression was not inhibited. Additionally, analysis utilizing protein synthesis inhibitor cycloheximide demonstrated that pHi mediated inhibition of COX-2 mRNA expression requires de novo protein synthesis of regulatory protein(s). These data strongly suggest that an alkaline pHi is an important trigger for constitutive COX-2 expression. Defining pHi -mediated mechanisms that govern the constitutive COX-2 expression may help in developing new strategies to block COX-2 over-expression in cancer cells. J. Cell. Physiol. 198: 295,301, 2004© 2003 Wiley-Liss, Inc. [source] PARTIAL PURIFICATION AND CHARACTERIZATION OF NEUTRAL TREHALASE FROM COMMERCIAL BAKER'S YEAST, SACCHAROMYCES CEREVISIAEJOURNAL OF FOOD BIOCHEMISTRY, Issue 6 2000SANIYE YARAR ABSTRACT The neutral trehalase of a commercial baker's yeast (S. cerevisiae) strain has been partially purified using ammonium sidfate fractionation and DEAE-cellulose column chromatography techniques. Trehalase was precipitated between 35,50% ammonium sulfate saturation and approximately 5,8 fold purification was achieved. The yeast cAMP-dependent protein kinase was also precipitated in the same fraction and these two proteins were separated by DEAE-cellulose column chromatography. Trehalase became totally inactive after ion exchange chromatography, "cryptic trehalase" (tre-c), but was later activated with the addition of partially purified protem kinase together with cAMP and ATP. A 215 fold purification was obtained after DEAE-ceUulose column chromatography. One mM EDTA caused complete inhibition of the enzyme in crude extract, however the inhibition levels in ammonium sulfate and DEAE-cellulose fractions were 73.5% and 50%, respectively. Optimal pH range and temperature of the enzyme were determined as pH 6,6.8 and 30C, respectively. The kinetic parameters, Km and Vmax, were estimated as 11.78 mM trehalose and 12.47 ,mole glucose/min-mg protein, respectively. [source] EFFECT OF ETHANOL VAPOR ON GROWTH AND TOXIN PRODUCTION BY CLOSTRIDIUM BOTULINUM IN A HIGH MOISTURE BAKERY PRODUCTJOURNAL OF FOOD SAFETY, Issue 2 2000DAPHNE PHILLIPS DAIFAS ABSTRACT To determine the effect of ethanol vapor on toxin production by Clostridium botulinum, studies were done in English style crumpets (aw 0.990, pH 6.5) challenged with 500 spores/g C. botulinum types A and proteolytic B and packaged in high gas barrier bags [ethanol transmission rate (ETR) 0.21 g/m2/day @ 25 C]. Crumpets were packaged in air with either commercially available ethanol vapor generators (Ethicap® 2, 4 or 6G) or cotton wool pads saturated with 2, 4 or 6 g of 95% food grade ethanol and stored at 25C. Toxin was detected in all inoculated control crumpets (0% ethanol) after 5 days at ambient temperature (25C). Ethicap® 2G delayed toxicity for 10 days while complete inhibition (>21 days) was observed in all crumpets packaged with 4 or 6G Ethicap® or with 2, 4 or 6 g of ethanol per pad. However, all crumpets were overtly spoiled by this time. Both headspace ethanol and absorption of ethanol by crumpets increased as a function of Ethicap® size/weight of ethanol. Based on these preliminary studies, ethanol vapor would appear to be an effective additional barrier to control the growth and toxin production by C. botulinum in high moisture bakery products and ensure the safety of these products at ambient temperature. [source] A Survey on the Potential Mode of Inhibition for Oxalic Acid on Polyphenol OxidaseJOURNAL OF FOOD SCIENCE, Issue 8 2003R. Yoruk ABSTRACT: The potential mode of inhibition for xalic acid on polyphenol oxidase (PPO) was investigated. The extent of inhibition was influenced not only by oxalic acid concentration but also by pH. Inhibition was most prominent at pH 4.0 where complete inhibition occurred at the 4-mM oxalic acid concentration and was less evident at higher pH values. Inhibition of PPO by oxalic acid was due to its binding with copper to form an inactive complex, and the inhibition was characterized as noncompetitive. Oxalic acid diminished the catechol-quinone product formation, and no quinone bleaching was observed. Oxalic acid was a more potent inhibitor of PPO compared with other structurally related acids. [source] Protective effect of rebamipide on indomethacin-induced intestinal damage in ratsJOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, Issue 10 2001Hiroyuki Mizoguchi Abstract Background and Aim: We evaluated the effect of rebamipide (2-(4-chlorobenzoylamino)-3-[2(1H)-quinolinon-4-yl] propionic acid), a novel anti-ulcer drug, on indomethacin-induced small intestinal lesions in rats. Methods: The animals were administered indomethacin (10 mg/kg, s.c.), and they were killed 24 h later. Rebamipide (30,300 mg/kg) was administered p.o. twice, 30 min before, and 6 h after indomethacin. Results: Indomethacin caused hemorrhagic lesions in the rat small intestine, accompanied by an increase in enterobacterial translocation, inducible nitric oxide synthase (iNOS) and myeloperoxidase (MPO) activities, as well as thiobarbituric acid (TBA) reactants, and these changes were significantly prevented by the supplementation with 16,16-dimethyl prostaglandin E2 (dmPGE2; 10 ,g/kg, i.v.) or the pretreatment of animals with the antibiotic ampicillin. Treatment of the animals with rebamipide dose-dependently prevented the development of intestinal lesions, and this effect was mimicked by i.v. administration of superoxide dismutase (SOD: 3000 U/kg) + catalase (CAT: 5000 U/kg). The protection by rebamipide was accompanied by a significant suppression of the increase in both MPO and iNOS activities, and a complete inhibition of the increase in TBA reactants, while SOD + CAT significantly inhibited the increase of MPO activity and TBA reactants, but not iNOS activity. The bacterial translocation following indomethacin was also significantly decreased by either rebamipide or SOD + CAT. Conclusion: These results confirmed the importance of enterobacteria and iNOS/NO in the pathogenesis of indomethacin-induced small intestinal lesions, and suggested that rebamipide prevents the development of these lesions, probably by its radical scavenging action. [source] Poster Session BP07: Neurodegenerative DiseasesJOURNAL OF NEUROCHEMISTRY, Issue 2002F. Jayman Presynaptic terminals contain an abundant 140-amino acid phosphoprotein, dubbed ,-synuclein, which is accumulated in Lewy bodies typically observed in neurons in neurodegenerative diseases, such as Parkinson's disease. In this study, the role of ,-synuclein in regulating cycle, differentiation, and survival of neuronal cells was studied using a rat dopaminergic cell line ZN27D. To delineate specific effects of ,-synuclein the same cell line was engineered to express human ,-synuclein and a vector-transfected cell line RK27 was used as a second control. All three cell lines showed significant proliferation even in serum-free medium, and complete inhibition of cell division and differentiation could be achieved in the ZN27D cells only when both dibutyryl cAMP (dbcAMP) and retinoic acid were present. In contrast, the ,-synuclein expressing cells could be differentiated in the presence of only dbcAMP. Dose dependence of MPP+(1-methyl-4-phenylpyridinium iodide)-mediated caspase3 activation was studied in undifferentiated ZN27D cells. At 200 ,m MPP+ a significant cleavage of the caspase3 substrate PARP was observed and it was reversed in the presence of ,-synuclein. MPP+ also inhibited aminophospholipid translocase (APTL), a P-type ATPase that is responsible for inner plasma membrane localization of phophotidylserine in healthy cells. The role of ,-synuclein in regulating cell cycle, differentiation, APTL activity and cell death is being investigated further in the dopaminergic ZN27D cell line. [source] Loss of lipopolysaccharide-induced nitric oxide production and inducible nitric oxide synthase expression in scrapie-infected N2a cellsJOURNAL OF NEUROSCIENCE RESEARCH, Issue 2 2003Heléne Lindegren Abstract In scrapie-infected cells, the conversion of the cellular prion protein to the pathogenic prion has been shown to occur in lipid rafts, which are suggested to function as signal transduction platforms. Neuronal cells may respond to bacterial lipopolysaccharide (LPS) treatment with a sustained and elevated nitric oxide (NO) release. Because prions and the major LPS receptor CD14 are colocalized in lipid rafts, the LPS-induced NO production in scrapie-infected neuroblastoma cells was studied. This study shows that LPS induces a dose- and time-dependent increase in NO release in the murine neuroblastoma cell line N2a, with a 50-fold increase in NO production at 1 ,g/ml LPS after 96 hr, as measured by nitrite in the medium. This massive NO release was not caused by activation of the neuronal NO synthase (nNOS), but by increased expression of the inducible NOS (iNOS) mRNA and protein. However, in scrapie-infected N2a cells (ScN2a), the LPS-induced NO production was completely abolished. The absence of LPS-induced NO production in ScN2a was due not to abolished enzymatic activity of iNOS but to a complete inhibition of the LPS-induced iNOS gene expression as measured by Western blot and RT-PCR. These results indicate that scrapie infection inhibits the LPS-mediated signal transduction upstream of the transcriptional step in the signaling cascade and may reflect the important molecular and cellular changes induced by scrapie infection. © 2002 Wiley-Liss, Inc. [source] Retinoic acid is a potential negative regulator for differentiation of human periodontal ligament cellsJOURNAL OF PERIODONTAL RESEARCH, Issue 6 2005Natsuko Shibuya Background and objectives:, Retinoic acid (RA) exerts a wide variety of effects on development, cellular differentiation and homeostasis in various tissues. However, little is known about the effects of RA on the differentiation of periodontal ligament cells. In this study, we investigated whether RA can affect the dexamethasone-induced differentiation of periodontal ligament cells. Methods and results:, Human periodontal ligament cells were differentiated via culturing in the presence of dexamethasone, ascorbic acid, and ,-glycerophosphate for mineralized nodule formation, as characterized by von Kossa staining. Continuous treatment with all- trans -RA inhibited the mineralization in a dose-dependent manner, with complete inhibition over 1 µm RA. Other RA analogs, 9- cis -RA and 13- cis -RA, were also effective. Furthermore, addition of RA for just the first 4 days completely inhibited the mineralization; however, as RA was added at later stages of culture, the inhibitory effect was diminished, suggesting that RA had a phase-dependent inhibition of mineralization. RA receptor (RAR)-, agonist (AM-580), but not retinoid X receptor agonist (methoprene acid), inhibited the mineralization, and reverse transcription,polymerase chain reaction analysis revealed that RAR-, was expressed on the cells, suggesting that RAR-, was involved in the inhibitory mechanism. This inhibition was accompanied by inhibition of alkaline phosphatase activity; however, neither expression of platelet-derived growth factor (PDGF) receptor-,, PDGF receptor-,, or epidermal growth factor (EGF) receptor, nor phosphorylation of extracellular signal-regulated kinases triggered by PDGF-ascorbic acid or PDGF-BB was changed, as assessed by flow cytometry or western blot analyses. Conclusions:, These findings suggest that RA is a potential negative regulator for differentiation of human periodontal ligament cells. [source] CONTRASTING EFFECTS OF METHIONINE SULFOXIMINE ON UPTAKE AND ASSIMILATION OF AMMONIUM IN ULVA INTESTINALIS (CHLOROPHYCEAE),JOURNAL OF PHYCOLOGY, Issue 4 2004Neill G. Barr Ammonium is assimilated in algae by the glutamine synthetase (GS),glutamine:2-oxoglutarate aminotransferase pathway. In addition to the assimilation of external ammonium taken up across the cell membrane, an alga may have to reassimilate ammonium derived from endogenous sources (i.e. nitrate reduction, photorespiration, and amino acid degradation). Methionine sulfoximine (MSX), an irreversible inhibitor of GS, completely inhibited GS activity in Ulva intestinalis L. after 12 h. However, assimilation of externally derived ammonium was completely inhibited after only 1,2 h in the presence of MSX and was followed by production of endogenous ammonium. However, endogenous ammonium production in U. intestinalis represented only a mean of 4% of total assimilation attributable to GS. The internally controlled rate of ammonium uptake (Vi) was almost completely inhibited in the presence of MSX, suggesting that Vi is a measure of the maximum rate of ammonium assimilation. After complete inhibition of ammonium assimilation in the presence of MSX, the initial or surge (Vs) rate of ammonium uptake in the presence of 400 ,M ammonium chloride decreased by only 17%. However, the amount that the rate of ammonium uptake decreased by was very similar to the uninhibited rate of ammonium assimilation. In addition, the decrease in the rate of ammonium uptake in darkness (in the absence of MSX) in the presence of 400 ,M ammonium chloride matched the decrease in the rate of ammonium assimilation. However, in the presence of 10 ,M ammonium chloride, MSX completely inhibited ammonium assimilation but had no effect on the rate of uptake. [source] UV-A/BLUE LIGHT,INDUCED REACTIVATION OF SPORE GERMINATION IN UV-B IRRADIATED ULVA PERTUSA (CHLOROPHYTA),JOURNAL OF PHYCOLOGY, Issue 2 2004Taejun Han Recent reduction in the ozone shield due to manufactured chlorofluorocarbons raised considerable interest in the ecological and physiological consequences of UV-B radiation (,=280,315 nm) in macroalgae. However, early life stages of macroalgae have received little attention in regard to their UV-B sensitivity and UV-B defensive mechanisms. Germination of UV-B irradiated spores of the intertidal green alga Ulva pertusa Kjellman was significantly lower than in unexposed controls, and the degree of reduction correlated with the UV doses. After exposure to moderate levels of UV-B irradiation, subsequent exposure to visible light caused differential germination in an irradiance- and wavelength-dependent manner. Significantly higher germination was found at higher photon irradiances and in blue light compared with white and red light. The action spectrum for photoreactivation of germination in UV-B irradiated U. pertusa spores shows a major peak at 435 nm with a smaller but significant peak at 385 nm. When exposed to December sunlight, the germination percentage of U. pertusa spores exposed to 1 h of solar radiation reached 100% regardless of the irradiation treatment conditions. After a 2-h exposure to sunlight, however, there was complete inhibition of germination in PAR+UV-A+UV-B in contrast to 100% germination in PAR or PAR+UV-A. In addition to mat-forming characteristics that would act as a selective UV-B filter for settled spores under the parental canopy, light-driven repair of germination after UV-B exposure could explain successful continuation of U. pertusa spore germination in intertidal settings possibly affected by intense solar UV-B radiation. [source] Acetic Acid, Ethanol and Steam Effects on the Growth of Botrytis cinerea in vitro and Combination of Steam and Modified Atmosphere Packaging to Control Decay in KiwifruitJOURNAL OF PHYTOPATHOLOGY, Issue 2 2009Anastasia L. Lagopodi Abstract The effects of acetic acid fumigation, ethanol fumigation, and steam heat treatment on growth of Botrytis cinerea in vitro were investigated. The effect of steam heat treatments in combination with modified atmosphere packaging (MAP) on Botrytis decay development on ,Hayward' kiwifruit was also studied. The fungus was grown in Petri dishes on potato dextrose agar. Ethanol fumigation with 100 ,l/l for 3 or 6 min, or 200 ,l/l for 6 min enhanced the growth of B. cinerea. The effect of acetic acid on growth of B. cinerea was time and dosage-dependent. Fumigation with 1 ,l/l for 6 min, 2 ,l/l for 3 min, and 4 ,l/l for 3 min promoted radial growth of the fungus when compared to the growth of the untreated control. Fumigation with 2 ,l/l for 6 min delayed the growth of the fungus for the first 6 days, while fumigation with 6 ,l/l for 3 min delayed the growth of the fungus after the sixth day. Fumigation with 4 or 6 ,l/l acetic acid for 6 min, and 8 ,l/l acetic acid for 3 or 6 min resulted in complete inhibition of fungal growth. Steam heat treatment at 45°C for 6 min, and at 48, 51, and 54°C for 3 or 6 min completely inhibited fungal growth in vitro. Furthermore, steam treatments at 47, 50, and 53°C for 3 or 6 min completely inhibited decay at the stem end of kiwifruit kept at 10°C in MAP for 12 days. However, none of the steam treatments inhibited decay in wounds on the surface of the fruit kept in MAP. [source] Scirpusin A, a hydroxystilbene dimer from Xinjiang wine grape, acts as an effective singlet oxygen quencher and DNA damage protectorJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 5 2010Qingjun Kong Abstract BACKGROUND: Grapes and red wines are rich sources of phenolic compounds such as anthocyanins, catechins, flavonols and stilbenes, most of which are potent antioxidants showing cardioprotective properties. We first isolated scirpusin A, a hydroxystilbene dimer, from a wine grape of Xinjiang, and studied its antioxidant activity. RESULTS: Reactive oxygen species scavenging effects and the protection against reactive singlet oxygen-induced DNA damage of scirpusin A have been investigated in our experiments. The concentration of scirpusin A required to inhibit 50% of 1O2 generation was 17 µmol L,1, while addition of scirpusin A at 140 µmol L,1 caused complete inhibition. Further kinetic study revealed that the reaction of Scirpusin A with singlet oxygen has an extremely high rate constant (ka = 4.68 × 109 L mol,1 s,1). Scirpusin A (140 µmol L,1) exhibited significant inhibition effects on pBR322 DNA breakage. However, scavenging effects of scirpusin A on superoxide anion O2,, and hydroxyl radical ·OH were not potent as the inhibitor rates at a concentration of 1400 µmol L,1 were 28.83% and 19.5%, respectively. CONCLUSION: The present study shows that scirpusin A is a selective quencher of singlet oxygen and a protector against reactive singlet oxygen-induced pBR322 DNA damage at very low concentrations. Copyright © 2010 Society of Chemical Industry [source] | |||||||||||||||||||||||||