Perturbation of oxidant-antioxidant status in non-obese prehypertensive male subjects.
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Adult↗
Biomedical subjects
N Selvaraj
Publications and source records attributed to N Selvaraj.
Vitamin E protects the insulin sensitivity and redox balance in rat L6 muscle cells exposed to oxidative stress.
BACKGROUND: The effect of reactive oxygen species (ROS) on insulin action is unclear. This study was carried out to explore the effect of prolonged low grade oxidative stress and vitamin E treatment on cultured rat L6 muscle cells. METHODS: L6 myotubes were incubated with vitamin E for 18 h and treated with H2O2 generating system for 12 h. Insulin stimulated glucose uptake, total antioxidant capacity and reduced glutathione concentration were measured. RESULTS: There was a marked decrease in insulin stimulated glucose transport in L6 cells exposed to H2O2 generating system. Pretreatment with vitamin E attenuates the effect of H2O2 on insulin action. Treatment with H2O2 caused a significant reduction in the levels of reduced glutathione and total antioxidant capacity, these alterations were reversed by vitamin E pretreatment. Vitamin E per se had no effect on insulin stimulated glucose transport in cells not exposed to oxidative stress. CONCLUSION: In conclusion, our observations indicate that vitamin E improves the free radical defense system potential and prevents oxidative stress induced insulin resistance in rat L6 muscle cells.
Animals↗
Evidence for the role of lipid peroxides on glycation of hemoglobin and plasma proteins in non-diabetic asthma patients.
BACKGROUND: Collective evidences reveal that malondialdehyde (MDA), reduced glutathione (GSH) and ascorbic acid can modulate protein glycation. We investigated the concentrations of MDA, GSH, ascorbic acid and protein glycation in asthma patients to delineate the possible association among these parameters. METHODS: Blood was collected from 18 asthma patients and 16 age and sex matched control subjects. Glycated hemoglobin (HbA1C), GSH, MDA, vitamin C, fructosamine and glucose were assessed in both groups. The effect of H2O2 on glycation of hemoglobin was studied by incubating normal healthy erythrocytes with either 5 or 50 mmol/l glucose concentration. RESULTS: Plasma of asthma patients revealed significantly higher concentrations of lipid peroxides and fructosamine concentrations than the matched controls. Glycated hemoglobin concentrations were also found to be significantly increased. Ascorbic acid and GSH concentrations were decreased significantly in the test group when compared with the healthy control group. When the effects of fasting glucose, GSH and ascorbic acid on the concentrations of HbA1C and fructosamine were refuted by partial correlation analysis, MDA was found to be a significant determinant of HbA1c and fructosamine in patients with asthma. The in vitro model with human erythrocytes showed an enhancement of protein glycation by H2O2. CONCLUSION: An increased glycation of proteins was found in asthma patients. These data also support the premise that lipid peroxides per se do have a role to play in glycation of hemoglobin and plasma proteins.
Adult↗
Effect of lipid peroxides and antioxidants on glycation of hemoglobin: an in vitro study on human erythrocytes.
BACKGROUND: Glycation and lipid peroxidation are two important processes known to play a key role in complications of many pathophysiological process. We sought to assess the possibility of an interaction between these processes in vitro and to examine the effect of lipoic acid and taurine on the glycation of hemoglobin and lipid peroxidation. METHODS: Human erythrocytes in phosphate buffered saline (pH 7.4) were incubated with 5 or 50 mmol/l glucose. To study the effect of antioxidants on glycation of hemoglobin, erythrocytes were incubated with either lipoic acid or taurine and then exposed to glucose concentration of either 5 or 50 mmol/l. To clarify if lipid peroxides per se enhances the glycated hemoglobin level, an in vitro study was performed by incubating erythrocyte suspension containing either 5 or 50 mmol/l glucose with or without MDA. Lipid peroxides and glycated hemoglobin levels were determined in the glucose treated cells. RESULTS: Glycated hemoglobin levels were higher in erythrocytes incubated with 50 mmol/l glucose concentrations than in erythrocytes incubated with 5 mmol/l glucose. The increase in glycated hemoglobin levels was blocked significantly when erythrocytes were pretreated with either lipoic acid or taurine. Both the antioxidants used in the present study markedly reduced the MDA levels. The level of glycated hemoglobin in erythrocyte incubated in the presence of MDA was increased significantly when compared to erythrocyte incubated with glucose alone. CONCLUSIONS: Lipid peroxides per se may have a role to play in glycation of hemoglobin and antioxidants (lipoic acid and taurine) can partially inhibit the formation of glycated hemoglobin by lowering the levels of lipid peroxides.
Antioxidants↗
Reassessing the increased glycation of hemoglobin in nondiabetic chronic renal failure patients: a hypothesis on the role of lipid peroxides.
BACKGROUND: Glycated hemoglobin (HbA(1C)) is considered clinically useful for assessing long-term integrated control of blood glucose in diabetes. However, an increased HbA(1C) concentration has been documented in chronic renal failure (CRF) patients without any history of diabetes. Collective evidences reveal that lipid peroxidation (MDA) can modulate protein glycation. We evaluated the relationship between glycated hemoglobin (HbA(1C)) and lipid peroxidation in non-diabetic CRF patients. METHODS: Twenty-eight nondiabetic CRF and 23 age- and sex-matched healthy subjects were enrolled for this study. Plasma urea, creatinine, lipid peroxides, fasting glucose and HbA(1C) were analyzed in both the groups. The in-vitro effect of MDA on glycation of hemoglobin was studied by incubating healthy erythrocytes with either 5 or 50 mmol/l glucose concentration. RESULTS: The percentage of HbA(1C) concentrations and plasma malondialdehyde (MDA) were significantly increased in CRF patients compared to control subjects. When the effects of uremia and blood glucose on the concentration of HbA(1C) was refuted by partial correlation analysis, MDA was found to be a significant determinant of HbA(1C) (r=0.41, p=0.04) in patients with renal failure. In-vitro incubation of RBC with glucose along with MDA was found to enhance the process of hemoglobin glycation. CONCLUSION: Our results suggest that lipid peroxidation per se can contribute to glycation of hemoglobin, warranting extra-precaution in interpreting HbA(1C) as a measure of glycemic control in CRF.
Adult↗
Agrobacterium-mediated genetic transformation and development of herbicide-resistant sugarcane (Saccharum species hybrids) using axillary buds.
Direct regeneration from explants without an intervening callus phase has several advantages, including production of true type progenies. Axillary bud explants from 6-month-old sugarcane cultivars Co92061 and Co671 were co-cultivated with Agrobacterium strains LBA4404 and EHA105 that harboured a binary vector pGA492 carrying neomycin phosphotransferase II, phosphinothricin acetyltransferase (bar) and an intron containing beta-glucuronidase (gus-intron) genes in the T-DNA region. A comparison of kanamycin, geneticin and phosphinothricin (PPT) selection showed that PPT (5.0 mg l(-1)) was the most effective selection agent for axillary bud transformation. Repeated proliferation of shoots in the selection medium eliminated chimeric transformants. Transgenic plants were generated in three different steps: (1) production of putative primary transgenic shoots in Murashige-Skoog (MS) liquid medium with 3.0 mg l(-1) 6-benzyladenine (BA) and 5.0 mg l(-1) PPT, (2) production of secondary transgenic shoots from the primary transgenic shoots by growing them in MS liquid medium with 2.0 mg l(-1) BA, 1.0 mg l(-1) kinetin (Kin), 0.5 mg l(-1) alpha-napthaleneacetic acid (NAA) and 5.0 mg l(-1) PPT for 3 weeks, followed by five more cycles of shoot proliferation and selection under same conditions, and (3) rooting of transgenic shoots on half-strength MS liquid medium with 0.5 mg l(-1) NAA and 5.0 mg l(-1) PPT. About 90% of the regenerated shoots rooted and 80% of them survived during acclimatisation in greenhouse. Transformation was confirmed by a histochemical beta-glucuronidase (GUS) assay and PCR amplification of the bar gene. Southern blot analysis indicated integration of the bar gene in two genomic locations in the majority of transformants. Transformation efficiency was influenced by the co-cultivation period, addition of the phenolic compound acetosyringone and the Agrobacterium strain. A 3-day co-cultivation with 50 micro M acetosyringone considerably increased the transformation efficiency. Agrobacterium strain EHA105 was more effective, producing twice the number of transgenic shoots than strain LBA4404 in both Co92061 and Co671 cultivars. Depending on the variety, 50-60% of the transgenic plants sprayed with BASTA (60 g l(-1) glufosinate) grew without any herbicide damage under greenhouse conditions. These results show that, with this protocol, generation and multiplication of transgenic shoots can be achieved in about 5 months with transformation efficiencies as high as 50%.
Acetophenones↗
Possible link between glycated hemoglobin and lipid peroxidation in hyperthyroidism.
BACKGROUND: Glycated hemoglobin (HbA1C) levels are enhanced by elevated glucose concentrations. Glycation of hemoglobin is also modulated by lipid peroxides, ascorbic acid and reduced glutathione (GSH). We determined the strength of the relationships among these variables in a group of hyperthyroid patients. METHODS: Twenty-two untreated hyperthyroid patients and 17 healthy controls were recruited for the study. Whole blood GSH, HbA1C, plasma lipid peroxides, ascorbic acid and fasting glucose were analyzed in both the groups. Direct and partial correlation analysis was performed to explore the possible relationships between these variables. RESULTS: In hyperthyroid patients, HbA1C and lipid peroxides levels were found to be significantly increased than the controls. Ascorbic acid and GSH were decreased significantly in the test group when compared with the healthy control group. With partial correlation analysis, fasting glucose and lipid peroxides were found to have a significant positive correlation with HbA1C. Ascorbic acid and GSH showed no significant association with HbA1C levels. CONCLUSION: These data suggest that HbA1C levels are closely associated with fasting glucose and lipid peroxides in hyperthyroid patients. Therefore, serum lipid peroxides level should be kept in mind while interpreting HbA1C as a long-term glycemic index in hyperthyroid cases.
Adolescent↗
Increased protein glycation in non-diabetic pediatric nephrotic syndrome: possible role of lipid peroxidation.
BACKGROUND: Malondialdehyde (MDA), ascorbic acid and reduced glutathione (GSH) have been reported to play a possible role in glycation of proteins. This study was performed to evaluate this correlation in nephrotic syndrome patients by comparing the levels of fructosamine with MDA, ascorbic acid and GSH. METHODS: Fifteen children with nephrotic syndrome during relapse and 10 age- and sex-matched healthy controls were enrolled for this study. Whole blood GSH, plasma MDA, total ascorbic acid and fasting glucose were analyzed in both the groups. Partial correlation analysis was performed to predict the independent association of MDA, ascorbic acid and GSH on fructosamine. RESULTS: Plasma MDA and fructosamine levels were found to be increased in nephrotic syndrome patients when compared with controls. Plasma ascorbic acid and whole blood GSH were decreased in nephrotic group vs. healthy controls. Partial correlation analysis showed a significant positive correlation between fructosamine and MDA. CONCLUSIONS: Present data point to a possible involvement of MDA in the glycation of protein in non-diabetic nephrotic syndrome patients, and provide support for the potential use of an antioxidant therapy in these patients.
Ascorbic Acid↗
An evaluation of level of oxidative stress and protein glycation in nondiabetic undialyzed chronic renal failure patients.
BACKGROUND: Chronic renal failure (CRF) patients on prolonged dialysis and with diabetes have been found to have significant alteration in their antioxidant status and protein glycation, but little is known about the same in nondiabetic undialyzed CRF patients. This study was performed to evaluate (a) the levels of oxidative stress and protein glycation in nondiabetic undialyzed CRF patients and (b) the possible influence of oxidative stress on protein glycation. DESIGN AND METHODS: A case control study was performed on 23 nondiabetic undialyzed CRF patients and 17 control subjects. The concentrations of total plasma ascorbic acid, whole blood reduced glutathione (GSH) and plasma lipid peroxides were measured to evaluate the antioxidant status and oxidative stress. The protein glycation was measured from plasma fructosamine normalized for albumin concentrations. A partial correlation analysis was carried out to analyze the effect of oxidative stress parameters on protein glycation. RESULTS: A compromised nonenzymatic defense against free-radical generation was evidenced by decreased concentrations of whole blood GSH and plasma ascorbic acid. The concentrations of lipid peroxides, fructosamine and fructosamine/albumin ratio were significantly (p<0.05) higher in the CRF group when compared with controls. The partial correlation analysis revealed that oxidative stress influences protein glycation in CRF patients. CONCLUSIONS: Increased oxidative stress might have a role in promoting protein glycation in nondiabetic undialyzed CRF patients. The reduction of oxidative stress and protein glycation might open new therapeutic approaches in treating CRF patients.
Adult↗
Differences in Ca(2+) signaling underlie age-specific effects of secretagogues on colonic Cl(-) transport.
Taurodeoxycholic acid (TDC) stimulates Cl(-) transport in adult (AD), but not weanling (WN) and newborn (NB), rabbit colonic epithelial cells (colonocytes). The present study demonstrates that stimuli like neurotensin (NT) are also age specific and identifies the age-dependent signaling step. Bile acid actions are segment and bile acid specific. Thus although TDC and taurochenodeoxycholate stimulate Cl(-) transport in AD distal but not proximal colon, taurocholate has no effect in either segment. TDC increases intracellular Ca(2+) concentration ([Ca(2+)](i)) in AD, but not in WN and NB, colonocytes. In AD cells, TDC (5 min) action on Cl(-) transport needs intra- but not extracellular Ca(2+). NT, histamine, and bethanechol increase Cl(-) transport and [Ca(2+)](i) in AD, but not WN, distal colonocytes. However, A-23187 increased [Ca(2+)](i) and Cl(-) transport in all age groups, suggesting that Ca(2+)-sensitive Cl(-) transport is present from birth. Study of the proximal steps in Ca(2+) signaling revealed that NT, but not TDC, activates a GTP-binding protein, Galpha(q), in AD and WN cells. In addition, although WN and AD colonocytes had similar levels of phosphatidylinositol 4,5-bisphosphate, NT and TDC increased 1,4,5-inositol trisphosphate content only in AD cells. Nonresponsiveness of WN cells to Ca(2+)-dependent stimuli, therefore, is due to the absence of measurable phospholipase C activity. Thus delays in Ca(2+) signaling afford a crucial protective mechanism to meet the changing demands of the developing colon.
Aging↗
Establishment and characterization of steroidogenic granulosa cells expressing beta(2)-adrenergic receptor: regulation of adrenodoxin and steroidogenic acute regulatory protein by adrenergic agents.
Primary granulosa cells obtained from PMSG primed immature rats were triple transfected with SV40 DNA, Ha-ras oncogene and an expression vector containing human beta(2)-adrenergic receptors resulting in granulosa cell lines constitutively expressing the beta(2)-adrenergic receptors. Isoproterenol, a potent adrenergic agent, stimulated both cAMP accumulation and progesterone production in these cells in a dose dependent manner. Responsiveness of these cells was specific only to isoproterenol, while hCG (2.4 nM) and hFSH (2.4 nM) had no effect on steroid production. ED(50) for stimulation of cAMP and progesterone in these cells by isoproterenol was 2x10(-6) M and 7x10(-6) M, respectively. Forskolin also showed a dose dependent stimulation of cAMP and progesterone with ED(50) of 1.5 and 0.35 microg/ml, respectively. Epinephrine at a dose of 10(-5) M elicited maximum response to produce cAMP and progesterone. Isoproterenol induced accumulation of cAMP and progesterone in these cells were inhibited by beta(2)-adrenergic blocker, propranolol with an ED(50) of 6x10(-8) and 7x10(-9) M, respectively, whereas the beta(1)-adrenergic blocker, metoprolol was effective only at a very high concentration (ED(50)>10(-4) and 1.9x10(-5) M for inhibiting isoproterenol induced cAMP and progesterone production, respectively). Induction of steroidogenesis by isoproterenol or forskolin involved de novo synthesis of the cytochrome P450 side chain cleavage (SCC) enzyme complex, as assessed by indirect immunofluorescence staining for adrenodoxin. Western analysis indicate that expression of adrenodoxin is upregulated by forskolin, isoproterenol and adrenalin by 7.8-, 6.9- and 10.8-fold, respectively. The presence of StAR protein was identified by Western blotting. StAR expression was elevated by 8.3-, 2.5- and 4.7-fold upon stimulation with forskolin, isoproterenol and adrenalin, respectively. Thus, this cell line could serve as a good model system to study catecholamine mediated regulation of growth and differentiation of granulosa cells and the role of oncogenes in this process.
1-Methyl-3-isobutylxanthine↗
Myocardial infarction, seizure and an abnormal head CT scan.
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Bupropion↗
Hemiparesis following influenza vaccination.
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Aged↗
Modulation of FSH receptor phosphorylation correlates with hormone-induced coupling to the adenylate cyclase system.
The authors have recently demonstrated that an inhibitor of protein phosphorylation, staurosporine (SSP), can dramatically enhance follicle-stimulating hormone (FSH) stimulated cyclic adenosine monophosphate (cAMP) accumulation in rat granulosa cell line (GFSHR-17) overexpressing about 20-fold FSH receptor than primary granulosa cells. Moreover, incubation with SSP can partially release the cells from FSH-induced desensitization. In this work, it was examined whether coupling of FSH receptor to the adenylate cyclase is correlated with the degree of receptor phosphorylation. Immunoprecipitation of FSH receptor after metabolic labeling of the cells with 32P-orthophosphate revealed that preincubation of the cells with SSP resulted in pronounced reduction in FSH receptor phosphorylation compared to control cells, concomitantly with a dramatic increase in FSH-stimulated cAMP accumulation. In contrast, incubation of the cells with saturating dose of FSH, which leads to uncoupling between the receptor and the adenylate cyclase, resulted in enhanced receptor phosphorylation. Moreover, cells preincubated with FSH could be released from desensitization by further incubation with SSP and a significant reduction in FSH receptor phosphorylation. Immunostaining of the cells with FSH receptor antibody reveal a homogenous distribution of the receptor on the surface of SSP-treated cells. Some aggregation of the receptor was evident in control cells that were not treated with SSP. In contrast, massive clustering and capping of the receptor molecules were observed on the surface of FSH-stimulated cells. The current data suggest that phosphorylation-dephosphorylation of the receptor molecules play an important role in the degree of coupling between the receptor and the adenylate cyclase system. Moreover, desensitization to FSH stimulation that is implicated with high degree of receptor phosphorylation may lead to aggregation of the receptor molecules on the cell surface.
Adenylyl Cyclases↗
Apoptosis in steroidogenic cells: structure-function analysis.
Granulosa cells are the main producers of the female sex steroid hormones, progesterone and estradiol, which are responsible for the cyclicity in ovarian function. Programmed cell death in the ovary plays a crucial role in limiting the number of follicles that can ovulate and thus prevents the development of more embryos than can successfully complete pregnancy. Granulosa cell apoptosis is regulated by the concerted action of endocrine, paracrine, and autocrine factors. These factors lead to the developmental decision of whether the steroidogenic cell will luteinize and enter the pathway leading to programmed cell death, or whether the life span of the luteinized cell will be prolonged to continue secretion of progesterone, which is essential for the maintenance of pregnancy. At the level of the individual cell, we find that enhanced steroidogenesis can be maintained during the initial steps of apoptosis as long as the steroidogenic apparatus remains intact. This can be achieved by a unique mechanism of compartmentalization of steroidogenic organelles in the perinuclear region and migration of the multicatalytic proteinase, the proteasome, to the apoptotic blebs. Reorganization of the actin cytoskeleton during apoptosis may provide an efficient barrier between the proteolytic activity and the steroidogenic activity in the apoptotic cell. It is suggested that steroidogenesis can be maintained in the apoptotic cells as long as the steroidogenic organelles bearing the steroidogenic apparatus remain intact.
8-Bromo Cyclic Adenosine Monophosphate↗
Control of differentiation, transformation, and apoptosis in granulosa cells by oncogenes, oncoviruses, and tumor suppressor genes.
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Animals↗
Partial sequencing of the rat steroidogenic acute regulatory protein message from immortalized granulosa cells: regulation by gonadotropins and isoproterenol.
Steroidogenic acute regulatory protein (StAR), a 30-kDa protein involved in the transport of cholesterol to inner mitochondrial membrane during stimulation of steroid hormone biosynthesis, has recently been cloned from human adrenals and MA-10 mouse Leydig tumor cells. We examined the regulation of StAR mRNA accumulation upon induction of steroidogenesis in immortalized rat granulosa cells. Granulosa cells were transfected with SV40 DNA alone (POGS5); with SV40 DNA and Ha-ras oncogene (POGRS1); with SV40 DNA, Ha-ras oncogene and LH/CG receptor (GLHR15) or with FSH receptor (GFSHR17) or with the beta 2-adrenergic receptor (G beta 2AR13) expression plasmids. Cells were cultured to confluency and then stimulated for 24 h with oFSH (4 nM), hCG (2.4 nM), isoproterenol (10 microM) or forskolin (50 microM). By quantitative RT-PCR, StAR mRNA was undetectable in non-steroidogenic cells (transfected with SV40 DNA alone, POGS5) either in the presence or in the absence of forskolin. In contrast, variable amount of the message was detected in all steroidogenic cell lines cotransfected with SV40 DNA and Ha-ras. Moreover, an increase in the StAR mRNA expression was evident in all steroidogenic cells upon stimulation with their respective agonists, concomitantly with enhanced progesterone production. The RT-PCR product was sequenced and the 379 base pairs of rat StAR were found to be 93% and 86% identical to mouse and human cDNA, respectively. The deduced 126 amino acid sequence was 95%, 88% and 88% identical to the mouse, human and bovine deduced protein sequences. We conclude that StAR message is expressed only in the steroidogenic rat granulosa cells and can be upregulated by FSH, hCG, isoproterenol and forskolin in the appropriate cell lines. In addition, we find that the rat StAR cDNA exhibit a high degree of homology with the mouse and human sequences.
Amino Acid Sequence↗
Expression of Ad4-BP/cytochrome P450 side chain cleavage enzyme and induction of cell death in long-term cultures of human granulosa cells.
Freshly isolated granulosa cells obtained from women undergoing in-vitro fertilization (IVF) become refractory to further gonadotrophin stimulation in culture due to their previous hormonal treatment. However, when precultured for 7 days in gonadotrophin-free medium they regain their response to both human chorionic gonadotrophin (HCG) and follicle stimulating hormone (FSH) with a 10-fold and 5-fold increase in progesterone production respectively, within an additional 7 days of culture. Forskolin, a direct activator of adenylate cyclase, increased progesterone levels 12-fold compared with non-stimulated cultures. Oestradiol formation was also significantly elevated (P < 0.005) following 48 h stimulation with luteinizing hormone (LH), FSH or forskolin. Intracellular cAMP levels rose 1.5-fold, 10-fold and 15-fold after 1 h of FSH, HCG or forskolin treatment. Expression of both cytochrome P450 side chain cleavage enzyme (SCC) and the steroidogenic transcription factor SF1/Ad4BP could be demonstrated by Western blotting. However, elevation of P450 SCC alone was evident following FSH and HCG stimulation. In the presence of serum, the ultrastructure of these cultured cells displayed numerous lipid droplets and well-developed mitochondria, characteristic of highly steroidogenic cells. The proportion of apoptotic nuclei in these cultures was < 30%. Removal of the serum increased apoptotic incidence to 40%, whereas addition of FSH prevented cell death significantly (P < 0.01). HCG and forskolin increased apoptosis to approximately 50%, while treatment with 8Br-cAMP led to 80% cell death. Our data suggest that, after prolonged culture, human granulosa cells can regain cAMP and steroidogenic response to gonadotrophin stimulation. Moreover, our experiments indicate that apoptosis and steroidogenesis can coexist in the same cell population while the interrelationship between these processes can be determined by the intracellular levels of cAMP.
Apoptosis↗