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Biomedical subjects

K A Balasubramanian

Publications and source records attributed to K A Balasubramanian.

At least 19 recordsLinked to original sources
PubMed2006

Intestinal mucosal alterations in rats with carbon tetrachloride-induced cirrhosis: changes in glycosylation and luminal bacteria.

Spontaneous bacterial peritonitis is a major cause of mortality after liver cirrhosis. Altered permeability of the mucosa and deficiencies in host immune defenses through bacterial translocation from the intestine due to intestinal bacterial overgrowth have been implicated in the development of this complication. Molecular mechanisms underlying the process are not well known. In order to understand mechanisms involved in translocation of bacteria, this study explored the role of oxidative stress in mediating changes in intestinal mucosal glycosylation and luminal bacterial content during cirrhosis. CCl4-induced cirrhosis in rats led to prolonged oxidative stress in the intestine, accompanied by increased sugar content of both intestinal brush border and surfactant layers. This was accompanied by changes in bacterial flora in the gut, which showed increased hydrophobicity and adherence to the mucosa. Inhibition of xanthine oxidase using sodium tungstate or antioxidant supplementation using vitamin E reversed the oxidative stress, changes in brush border membrane sugar content, and bacterial adherence. In conclusion, oxidative stress in the intestine during cirrhosis alters mucosal glycosylation, accompanied by an increased hydrophobicity of luminal bacteria, enabling increased bacterial adherence onto epithelial cells. This might facilitate translocation across the mucosa, resulting in complications such as spontaneous bacterial peritonitis.

Animals↗
PubMed2005

Indomethacin induces free radical-mediated changes in renal brush border membranes.

Nonsteroidal anti-inflammatory drugs (NSAIDs) are used extensively in clinical medicine. One disadvantage of their use, however, is the occurrence of adverse effects in the kidneys. The side effects produced in this organ have been classically attributed to the inhibitory effect of these drugs on the activity of cyclooxygenase, a key enzyme in prostaglandin synthesis. Our earlier work with indomethacin, a commonly used NSAID, has shown that oxidative stress and mitochondrial dysfunction occur in the kidney in response to the drug. In view of this, this study looked into the effect of indomethacin on brush border membranes (BBM) from the kidney, as these biomembranes are prime targets of oxygen free radicals. Rats, fasted overnight, were dosed with indomethacin (20 mg/kg) by gavage and sacrificed 24 h later. BBM were isolated from the kidneys by polyethylene glycol precipitation. It was found that there was an increase in levels of products of peroxidation and a fall in the level of alpha-tocopherol in the BBM from indomethacin-dosed rats. These BBM also exhibited impaired glucose transport. The lipid composition of the membranes was also found to be altered. Alterations in lipids were associated with up-regulation of phospholipase A2. Pretreatment with L-arginine, a nitric oxide donor, protected against these effects of indomethacin. Thus, this study suggests that indomethacin induces impairment in structure and function of BBM in the kidney, with these effects possibly mediated by free radicals and activation of phospholipases. We postulate that such alterations may be important in the pathogenesis of NSAID-induced nephropathy.

Administration, Oral↗
PubMed2005

Surgical stress-induced alterations in retinoid metabolism in the small intestine: role of oxygen free radicals.

Oxidative stress in the small intestine can result in altered cell proliferation, migration, and differentiation of villus-crypt cells. Retinoid metabolism is recognized as an important mediator of cellular differentiation in the intestine. This study examined the effect of oxidative stress in retinoid metabolism in a surgical stress model. Surgical stress was performed by handling the intestine as done during laparotomy. Villus-crypt cells were isolated at different time periods and various retinoid concentrations in the cell homogenate and the retinoic acid forming enzymes were quantitated using HPLC. Surgical stress resulted in altered retinoid levels in various cell populations in the small intestine at 1 and 12h. The activity of alkaline phosphatase and retinal oxidase was also altered at these time points and all these changes were prevented by inhibiting superoxide generation using xanthine oxidase inhibitor, allopurinol. These studies suggest that alterations seen in enterocytes during surgical stress may be mediated by changes in retinoid metabolism.

Aldehyde Oxidoreductases↗
PubMed2005

Surgical manipulation of the intestine and distant organ damage-protection by oral glutamine supplementation.

BACKGROUND: The intestine is increasingly recognized as a primary effector of distant organ damage, such as the lung, after any abdominal surgery. Earlier studies have shown that surgical manipulation of the intestine induces generation of reactive oxygen species in the intestine, resulting in mucosal and lung damage. Because glutamine is preferentially used by the small intestine as an energy source, this study examined the effect of glutamine and glutamic acid on intestinal and lung damage after surgical manipulation. METHODS: Controls and rats were pretreated for 7 days with 2% glutamine or glutamic acid, or the isonitrogenous amino acids glycine or alanine in the diet and subjected to surgical manipulation of the intestine. The intestine and lung were assessed for damage, and protection offered by various amino acids was studied. RESULTS: Surgical manipulation resulted in oxidative stress in the intestine as evidenced by increased xanthine oxidase activity and decreased antioxidant status. Enterocyte mitochondria were also functionally impaired with altered calcium flux, decreased respiratory control ratio, and increased swelling. Gut manipulation also resulted in neutrophil infiltration and oxidative stress in the lung as assessed by an increase in myeloperoxidase activity, lipid peroxidation, and antioxidant status. Glutamine or glutamic acid supplementation for 7 days before surgical manipulation showed a protective effect against the intestinal and lung damage. CONCLUSIONS: This study suggests that preoperative enteral glutamine or glutamic acid supplementation attenuates intestinal and lung damage in rats during surgical manipulation and that this effect might offer protection from postsurgical complications.

Administration, Oral↗
PubMed2004

Altered glycosylation of surfactant and brush border membrane of the small intestine in response to surgical manipulation.

BACKGROUND: Surgical stress can lead to bacterial translocation from the intestine into systemic circulation. Adherence of bacteria onto the glycoconjugates of the brush border membrane (BBM) and surfactant coat (SLP) of the mucosal cells is the first step in the translocation of luminal bacteria. Our earlier study showed that surgical manipulation of the intestine results in oxidative stress leading to structural and functional alterations in the mucosa. This study looks at the effect of surgical manipulation on the glycoconjugate alterations of SLP and BBM. MATERIALS AND METHODS: SLP and BBM were isolated from control and after surgical manipulation and the sugar composition was analyzed. Bacterial adherence using E. coli isolated from cecum was compared after coating microtiter plates with SLP or BBM isolated from control and after surgical manipulation. RESULTS: An increase in various sugars was seen after surgical manipulation both in SLP and BBM and this increase was maximum at 12 h after surgery. These alterations increased bacterial adherence onto SLP and BBM. Inhibiting superoxide generation by allopurinol treatment prior to surgical manipulation prevented glycosylation alteration and bacterial adherence. CONCLUSION: Surgical manipulation results in altered glycoconjugates of SLP and BBM which leads to increased bacterial adherence. These alterations are probably brought about by oxygen-free radicals. This is clinically significant because postsurgical complications such as sepsis may be brought about by altered glycosylation.

Allopurinol↗
PubMed2003

Heat preconditioning prevents oxidative stress-induced damage in the intestine and lung following surgical manipulation.

BACKGROUND: The intestine is increasingly recognized as a primary effector of distant organ damage, such as lung, following abdominal surgery. Surgical manipulation of the intestine generates oxygen free radicals resulting in mucosal damage. Heat preconditioning has been proposed to prevent various stress-induced alterations in cells and tissues, including oxidative stress. This study examined the effect of heat preconditioning on oxidative stress-induced damage to the intestine and lung, following surgical manipulation. METHODS: Control rats and rats pretreated with heat were subjected to surgical manipulation by opening the abdominal wall and handling the intestine as done during laparotomy. Intestine and lung were assessed for damage by histology and markers of oxidative stress. RESULTS: Surgical manipulation resulted in ultrastructural changes in the intestine. Biochemical alterations in the enterocytes were evident, with increased xanthine oxidase activity resulting in production of superoxide anion and with a decrease in antioxidant status. Gut manipulation also resulted in neutrophil infiltration and oxidative stress in the lung as assessed by histology, myeloperoxidase activity, lipid peroxidation and antioxidant status. Heat conditioning before surgical manipulation had a protective effect against this intestinal and lung damage. CONCLUSION: This study suggests that mild whole-body hyperthermia before surgery might offer protection from postoperative complications.

Animals↗
PubMed2003

A simple method of rat renal brush border membrane preparation using polyethylene glycol precipitation.

A simple method for preparation of brush border membranes (BBM) from rat kidney using polyethylene glycol (PEG) precipitation has been described. This method avoids the use of cations for the preparation, which might alter membrane lipid composition. These preparations were assessed for enrichment of marker enzymes, contamination by subcellular structures, lipid composition and transport function. An enrichment of 11.8910-fold of alkaline phosphatase, 13.9500-fold of amino peptidase and 13.6500-fold of gamma-glutamyl transpeptidase and an approximate yield of 60% were seen in the final membrane preparation as compared to the homogenate. There was very little contamination of basolateral membranes, peroxisomes, microsomes or lysosomes in the final membrane preparation. Analysis of sugars indicated high content of fucose and sailic acid as compared to hexoses. Isolated membranes appeared as vesicles as seen by electron microscopy. Lipid analysis indicated the presence of various neutral and phospholipids with a high content of sphingomyelin along with a cholesterol/phospholipid ratio of 0.4850. The isolated membrane vesicles were able to transport glucose. This study has shown a simple method of renal brush border membrane preparation, which is comparatively pure and functionally active.

Animals↗
PubMed2003

Effect of oxidants on small intestinal brush border membranes and colonic apical membranes--a comparative study.

This study compares composition of the rat small intestinal brush border membranes (BBM) and colonic apical membranes (CAM) and their susceptibility to in vitro exposure to various oxidants. Differences were observed between BBM and CAM in their lipid composition, sugar content, alkaline phosphatase (ALP) activity and cholesterol/phospholipid ratio. BBM and CAM were exposed to superoxide generated by xanthine+xanthine oxidase (X-XO) or peroxides such as tertiary butyl hydroperoxide (tBuOOH) and hydrogen peroxide (H(2)O(2)) and alterations in ALP activity, peroxidation parameters and membrane lipids were analyzed. Exposure of BBM and CAM to superoxide resulted in decrease in ALP activity and increase in peroxidation parameters such as protein carbonyl content, malondialdehyde and conjugated diene. Superoxide exposure also resulted in lipid alterations specifically in certain phospholipids. These alterations were prevented either by superoxide dismutase or by allopurinol. Peroxides did not have any significant effect. These results suggest that both BBM and CAM are susceptible to superoxide, which can bring about peroxidation and degradation of membrane lipids specifically, certain phospholipids.

Alkaline Phosphatase↗
PubMed2003

Isolation of human small intestinal brush border membranes using polyethylene glycol and effect of exposure to various oxidants in vitro.

This study presents a method of brush border membrane (BBM) preparation from the human small intestine using polyethylene glycol (PEG) precipitation and also looks at the effect of in vitro oxidant exposure on structural and functional alterations in the membrane. Isolated BBM were relatively pure as judged by 10- to 14-fold enrichment of marker enzymes with less than 1% contamination by other subcellular organelles. These membranes showed uphill transport of glucose and lipid analysis showed a cholesterol-phospholipid (C/P) ratio of 1.19. Isolated BBM were found to be susceptible to superoxide generated by xanthine oxidase (XO), resulting in lipid and protein oxidation along with altered glucose uptake. Superoxide exposure also resulted in phospholipid alterations, especially generation of lyso phospholipids. These changes were prevented by inhibiting XO by allopurinol or scavenging superoxide by superoxide dismutase (SOD). Other oxidants studied did not have significant affect on these membranes. These studies suggest that PEG can be used for preparation of BBM from the human small intestine and these membranes undergo structural and functional alterations on exposure to superoxide.

Alkaline Phosphatase↗
PubMed2002

Surgical manipulation of the small intestine and its effect on the lung.

BACKGROUND: Surgical manipulation of the intestine results in generation of oxygen free radicals leading to mucosal damage as evidenced by ultrastructural and biochemical changes. It is likely that the gut-derived mediators can bring about damage to distant organs such as the lung. METHODS: Surgical manipulation of the gut was performed by opening the abdominal wall and handling the intestine. Lung damage was assessed by histology, markers of oxidative stress, and protein content in bronchoalveolar lavage fluid. Protection offered by pretreatment with various compounds such as allopurinol, L-arginine, quinacrine, and indomethacin was also studied. RESULTS: Gut manipulation resulted in neutrophil infiltration, oxidative stress, and permeability changes in the lung and these changes were maximum 30 and 60 min following surgical manipulation, which recovered with time and reversed to normal by 24 h. Prior treatment with inhibitors of xanthine oxidase, phospholipase A(2), or cyclooxygenase showed a protective effect against lung damage. CONCLUSION: This study has shown that laparotomy and intestinal handling result in distant organ (lung) damage which is probably brought about by neutrophil infiltration and oxidative stress on the lung. This is likely mediated by compounds generated in the intestine and transported into the systemic circulation since inhibition of generation of chemical mediators in the intestine offers protection against lung damage.

Allopurinol↗
PubMed2002

Heat preconditioning attenuates oxygen free radical-mediated alterations in the intestinal brush border membrane induced by surgical manipulation.

BACKGROUND: The intestine is highly susceptible to free radical-induced damage and our earlier work has shown that surgical manipulation of the intestine results in generation of oxygen free radicals and mucosal damage along with alterations in the brush border membrane (BBM). Heat preconditioning is known to offer protection against various stresses including oxidative stress and this study looked at the effect of heat preconditioning on the intestinal BBM alterations following surgical manipulation. METHODS: Control and rats heat preconditioned were subjected to surgical manipulation by opening the abdominal wall and handling the intestine. BBM were isolated from the intestine and structural and functional alterations to these membranes were assessed. RESULTS: Surgical manipulation resulted in oxidative stress suggested by a decrease in alkaline phosphatase activity and alpha-tocopherol content, accompanied by an increase in lipid peroxidation. A decrease in glucose transport by the isolated BBM vesicles suggested functional impairment. Surgical manipulation resulted in phospholipid degradation with generation of arachidonic acid along with appearance of cPLA(2) in the membrane. These changes were prevented by heat preconditioning of the animal prior to surgical manipulation. CONCLUSION: These results suggest that heat preconditioning offers protection from damage to the intestinal BBM following surgical manipulation and mild whole body hyperthermia might prevent postsurgical complications.

Alkaline Phosphatase↗
PubMed2002

Heat preconditioning prevents enterocyte mitochondrial damage induced by surgical manipulation.

BACKGROUND: The small intestine is susceptible to free radical-induced damage and our earlier work has shown that surgical manipulation of the intestine results in generation of oxygen free radicals, leading to mucosal damage. Heat preconditioning has been shown to offer protection against various stresses including oxidative stress and this study looked at the effect of heat preconditioning on surgical manipulation-induced intestinal mitochondrial alterations. METHODS: Control and rats pretreated with heat were subjected to surgical manipulation by opening the abdominal wall and handling the intestine as done during laparotomy. Mitochondria were prepared from isolated enterocytes and structural and functional alterations were assessed. RESULTS: Surgical manipulation of the intestine resulted in mitochondrial alterations as seen by ultrastructural changes and altered lipid composition. Mitochondria were functionally impaired as evidenced by altered calcium flux, decreased respiratory control ratio, and increased tetrazolium dye reduction and swelling. Along with this, biochemical alterations such as increased lipid and protein oxidation were seen following surgical manipulation. Mild heat preconditioning of the animal prevented these damaging effects. CONCLUSIONS: These studies suggest that stress in the small intestine due to surgery can affect enterocyte mitochondrial structure and function and these effects can be prevented by mild whole body hyperthermia prior to surgery.

Animals↗
PubMed2001

A novel method of preparation of small intestinal brush border membrane vesicles by polyethylene glycol precipitation.

A novel method of brush border membrane vesicle (BBMV) preparation from the small intestinal mucosa using polyethylene glycol (PEG) precipitation has been presented. This preparation is compared with calcium-precipitated BBMVs in marker enzyme enrichment, contamination by other subcellular membranes, transport of glucose, and lipid composition. PEG-precipitated BBMVs are comparable with calcium-precipitated membranes in all parameters except lipid composition and thiol content. PEG-precipitated membranes have more phosphatidylcholine and phosphatidylethanolamine and less lysophosphatidylcholine and lysophosphatidylethanolamine as compared to calcium-precipitated membranes. Diacylglycerol and triacylglycerol content are also high in PEG-precipitated membranes. Alteration in lipid composition indicate the possible activation of lipase and phospholipase by calcium during BBMV preparation, which is not seen in PEG precipitation. Thiol content is almost double in PEG-precipitated membranes as compared to calcium-precipitated membranes. These results indicate that PEG can be used for the preparation of BBMVs in native form from the intestine without any alteration in their structural components, and these membranes show comparable transport activity.

Alkaline Phosphatase↗
PubMed2001

Nitric oxide protects the intestine from the damage induced by laparotomy and gut manipulation.

BACKGROUND: The intestine is highly susceptible to free radical-induced damage, and our earlier work has shown that surgical stress induces the generation of oxygen free radicals in enterocytes, resulting in intestinal damage along with ultrastructural changes. Since nitric oxide (NO) is an important mediator of gastrointestinal function, this study looked at the effect of NO on surgical stress-induced intestinal alterations. MATERIALS AND METHODS: Control rats and rats pretreated with the NO donor l-arginine were subjected to surgical stress by opening the abdominal wall and handling the intestine as done during laparotomy. Enterocytes were isolated and homogenate prepared, and the protection offered by l-arginine against damage due to surgical stress was determined and compared with normal controls. Protection to structural as well as functional aspects of the intestine was also examined. RESULTS: Intestinal manipulation affected intestinal structure as assessed by electron microscopy. Functional impairment of the enterocyte was also evident, with increased xanthine oxidase activity resulting in production of superoxide anion. This impairment is more dramatic in the crypt cells. Increased protease activity was also seen following laparotomy and handling. Pretreatment with the NO synthase substrate l-arginine prevented these damaging effects. Arginine protection was abolished in the presence of the NO synthase inhibitor NG-nitro-l-arginine methyl ester, indicating the role of NO. CONCLUSION: Stress in the small intestine due to any surgery can affect enterocyte structure and function. These damaging effects can be prevented by NO, an important modulator of cellular function.

Animals↗
PubMed2001

Intestinal mitochondrial dysfunction in surgical stress.

BACKGROUND: Surgical stress is associated with altered intestinal function. Our earlier study using a rat model indicated that oxidative stress plays an important role in this process. Since mitochondria are crucial to cellular function and survival and are both a target as well as a source of reactive oxygen species, the present study looks at the changes in enterocyte mitochondria during surgical stress. METHODS: Surgical stress was induced by opening the abdominal wall and handling the intestine as done during laparotomy. Mitochondria were prepared from the isolated enterocytes at different time periods after surgical stress. The effect of surgical stress on enterocyte mitochondrial ultrastructure, respiration, anti-oxidant enzyme activity, thiol redox status, calcium flux, permeability, and matrix enzymes was then studied. RESULTS: Surgical stress resulted in alterations in mitochondrial respiration and thiol redox status. It was also associated with altered mitochondrial matrix enzyme activity, decreased superoxide dismutase activity, induction of mitochondrial permeability transition, and swelling, as well as impairment of mitochondrial calcium flux. These alterations were seen at a maximum of 60 min following surgical stress and were reversed by 24 h. CONCLUSIONS: Laparotomy and mild intestinal handling itself results in enterocyte mitochondrial damage. Since mitochondria are important cellular organelles, this damage can probably lead to compromised intestinal function.

Animals↗
PubMed2001

Nitric oxide prevents intestinal mitochondrial dysfunction induced by surgical stress.

BACKGROUND: The intestine is highly susceptible to free radical-induced damage and earlier work has shown that surgical stress induces generation of oxygen free radicals in enterocytes, resulting in intestinal damage along with changes in mitochondrial structure and function. Nitric oxide is an important mediator of gastrointestinal function and this study looked at the effect of nitric oxide on surgical stress-induced intestinal mitochondrial alterations. METHODS: Controls and rats pretreated with the nitric oxide donor L-arginine were subjected to surgical stress by opening the abdominal wall and handling the intestine. Enterocytes were isolated, mitochondria prepared and the protection offered by L-arginine against damage due to surgical stress was determined. Protection to structural as well as functional aspects of mitochondria was examined. RESULTS: Mild handling of the intestine affected the enterocyte mitochondrial structure as assessed by lipid composition and electron microscopy. Mitochondria were also functionally impaired with altered calcium flux and decreased respiratory control ratio. Pretreatment with the nitric oxide synthase substrate L-arginine prevented these damaging effects of surgical stress. Protection with arginine was abolished by the nitric oxide synthase inhibitor NG-nitro-L-arginine methyl ester, indicating the role of nitric oxide. CONCLUSION: Surgical stress in the small intestine can affect enterocyte mitochondrial structure and function. These damaging effects can be prevented by nitric oxide, an important modulator of cellular function.

Animals↗
PubMed2001

Intestinal dysfunction in liver cirrhosis: Its role in spontaneous bacterial peritonitis.

Spontaneous bacterial peritonitis is a common illness in patients with cirrhosis and ascites that occurs without any apparent focus of infection. Bacterial translocation plays an important role in spontaneous bacterial peritonitis and it is evident from a variety of studies that the gut is a major source of this bacteria. Gut motility alterations, along with bacterial overgrowth and changes in intestinal permeability, probably play a role in this bacterial translocation. The present review looks at the role of the intestine in spontaneous bacterial peritonitis induced by liver cirrhosis and the factors influencing bacterial translocation in this disease.

Animals↗