Anti-microbial activity of turmeric natural dye against different bacterial strains.

Curcumin, a yellow pigment present in the Indian spice turmeric (associated with curry powder), has been linked with suppression of inflammation; angiogenesis; tumorigenesis; diabetes; diseases of the cardiovascular, pulmonary, and neurological systems, of skin, and of liver; loss of bone and muscle; depression; chronic fatigue; and neuropathic pain. The utility of curcumin is limited by its color, lack of water solubility, and relatively low in vivo bioavailability. Because of the multiple therapeutic activities attributed to curcumin, there is an intense search for the ‘‘super curcumin’’. In the present study anti microbial activity of turmeric natural dye against different bacterial strains. In the present study in vitro, test confirmed the antimicrobial activity of turmeric extract against ten different bacterial strains. The antibacterial activity was measured by agar well diffusion method. The natural dye showed antibacterial activity against all the test bacterial isolates. Turmeric natural dye showed good inhibitory activity against E.coli and Vibrio cholera with a zone of inhibition 7mm to 15mm and 10mm to 15mm respectively. As turmeric powders are used to color natural fibers from which consumer products are manufactured. The incorporation of turmeric dye with natural fiber will help to produce value added handicrafts.

Effect of Bacillus subtilis PB6, a natural probiotic on colon mucosal inflammation and plasma cytokines levels in inflammatory bowel disease.

The pathophysiology of inflammatory bowel disease (IBD) involves the production of diverse lipid mediators, namely eicosanoid, lysophospholipids, and platelet-activating factor, in which phospholipase A2 (PLA2) is the key enzyme. Thus, it has been postulated that control of lipid mediators production by inhibition of PLA2 would be useful for the treatment of IBD. This hypothesis has been tested in the present study by examining the therapeutic effect of a novel natural probitic Bacillus subtilis PB6 (ATCC- PTA 6737). B. subtilis PB6 is found to secrete surfactins (cyclic lipopeptides) which have anti-bacterial potential. These surfactins inhibit PLA2, a rate-limiting enzyme involved in the arachidonic acid associated inflammatory pathway and could downregulate the inflammatory response by regulating the eicosanoid and cytokine pathways. With this concept, an experimental animal trial has been conducted in a rat model of 2, 4, 6-trinitrobenzene sulfonic acid (TNBS)-induced colitis. The oral administration of PB6 suppresses the colitis as measured by mortality rate, changes in the weight gain, colon morphology and the levels of plasma cytokines. The animals treated orally with PB6 at 1.5 x 10(8) CFU/kg thrice daily from day 4 to 10 significantly improve gross pathology of the colon and regain the colon weight to normal (p < 0.05), compared to TNBS-induced positive control. The plasma levels of pro-inflammatory cytokines (TNF-alpha, 1L-1beta, IL-6 and IFN-gamma) are also significantly lowered (p < 0.05) and anti-inflammatory cytokine (IL-I0 and TGF-beta) significantly (p < 0.05) increased after the oral administration of PB6 on day 11. The present study supports the concept that PB6 inhibits PLA2 by the secreting surfactins. In a clinical investigation, it is found to be well tolerated by all the healthy volunteers.

Effect of renal ischaemia reperfusion on calcium oxalate retention.

The effect of ischaemia reperfusion induced renal injury for calcium oxalate deposition under normal and simulated conditions was studied. Male Wistar rats of both control (group I) and urolithic (group II) groups underwent (1 h) unilateral renal artery occlusion and were subjected to 1, 3, 6, 12, 24 and 72 h reperfusion. The group I rats subjected to 1 h renal ischaemia followed by 3 and 6 h reperfusion had significant oxalate retention than that of sham operated controls. In group II, under hyperoxaluric condition, in addition to accumulation of oxalate, calcium oxalate deposits were also observed. The increased retention of calcium oxalate was attributed to increased oxalate binding protein activity, oxalate synthesizing enzymes lactate dehydrogenase and xanthine oxidase activities and accumulation of calcium. Our findings suggested that renal cellular injury produced by ischaemia reperfusion could accelerate calcium oxalate precipitation reaction.

Oxalate binding protein from the kidney of rat and human mitochondria: studies on properties.

The oxalate binding protein of rat and human kidney mitochondria were extracted by Triton X-100 and purified on Sephadex G-200 column followed by HPLC. Their molecular masses were found to be 62 kD and 58 kD respectively, rich with arginine and acidic amino acids, 7% of carbohydrates and 1% of inorganic ions. Antibodies raised to the rat protein inhibited the oxalate binding and cross-reacted to the human protein as well as rat liver protein. The binding of oxalate to the protein was rapid, reversible, dependent on concentration of oxalate, temperature sensitive and inhibited by oxalate analogues. The saturation reached at 175 nM oxalate for rat protein with a Kd of 33.3 nM and Bmax of 21 nmoles while for human protein the saturation reached at 183 nM oxalate and had a Kd of 41 nM and Bmax of 14 n moles. The half-saturation concentration of inhibitor (IC50) of oxalate was 0.25 microM for rat protein and 0.225 microM for human protein while the structural analogues of oxalate had higher IC50 values. Proteoliposomes showed accumulation of oxalate confirming transport function of the protein. The rat protein promoted calcium oxalate crystallization in vitro better than that of human protein and antibody inhibited the crystal growth in vitro.

Effect of vitamin E and mannitol on renal calcium oxalate retention in experimental nephrolithiasis.

The calcium oxalate stone formation is induced in rats by a single injection of sodium oxalate (i.p., 7 mg/100 g body weight). There was increase in kidney oxalate concentration and kidney mitochondrial oxalate binding activity with increased lipid peroxidation. Histopathological observations showed larger aggregates of calcium oxalate crystals in the renal tubules. At 12 hours after oxalate administration a maximal crystal deposition in the renal tubule with denuded epithelium, lymphocytic infiltration and blood were observed. Increased blood urea and creatinine indicated kidney failure after oxalate administration. Calcium oxalate crystalluria, hematuria, and proteinuria with casts were observed. Renal antioxidants vitamin E, ascorbic acid and glutathione were significantly decreased on oxalate challenge. Pretreatment with vitamin E provided only partial protection from calcium oxalate deposition. Pretreatment with vitamin E and mannitol together protected the renal tubules completely from calcium oxalate deposition by normalizing the tissue oxalate concentration and mitochondrial oxalate binding activity and increasing the concentration of antioxidants on oxalate challenge.

Liver function tests in recurrent P. vivax malaria.

Blood samples were collected from 61 P. vivax infected fresh and recurrent malaria patients and liver function parameters studied. Plasma albumin, A/G ratio were found decreased significantly (p < 0.001) when compared to controls. Among the group of recurrent malaria patients with more than five attacks lowest values were found and the decrease was directly correlated with the number of attacks. The enzyme activities of plasma LDH, SGPT and thymol turbidity were found increased significantly with the increase in the number of attacks (p < 0.001). The increase was more pronounced in more than 5 attack (R3) group. The levels of total, conjugated and free bilirubin and the enzyme activities of SGOT, alkaline phosphatase were also found increased significantly in all the recurrent malarial groups, when compared to controls, without any correlation between the number of attacks. The isoenzyme pattern of plasma LDH was not altered in either fresh or recurrent malarial attack groups when compared to controls.

Oxalate binding to rat kidney mitochondria: induction by oxidized glutathione.

Increased oxalate binding with negative correlation with reduced glutathione content was observed during lipid peroxidation in rat kidney mitochondria. In presence of oxidized glutathione (GSSG), peroxidized mitochondria lost 48% of protein-SH with concomitant 3-fold increase in oxalate binding activity while control mitochondria lost only 20% protein-SH with only 0.8 fold increase in oxalate binding activity. The GSSG-induced oxalate binding was apparently due to two-fold increased affinity of oxalate to the protein. Reduced glutathione (GSH) inhibited oxalate binding competitively with Ki, 1.4 x 10(-3) M. Urolithic rat kidney mitochondria showed 30-50% increase in oxalate binding activity along with depletion of GSH and protein-SH. These studies suggest that oxalate binding is regulated by thiol status of mitochondria.

Restoration of tissue antioxidants and prevention of renal stone deposition in vitamin B6 deficient rats fed with vitamin E or methionine.

Observed loss in body weight gain, increased lipid peroxidation reaction, decreased concentrations of antioxidants, ascorbic acid, alpha-tocopherol and reduced glutathione and antioxidant enzymes, glutathione peroxidase and catalase and increased concentration of hydroperoxides and hydroxyl radicals in vitamin B6 deficient rat liver [J Nutri Biochem, 2 (1991) 245] and kidney [Biochem International, 21 (1991) 599] were nearly normalized on feeding with vitamin E or methionine. Accumulation of oxalate and calcium during vitamin B6 deficiency was abolished by feeding vitamin E or methionine. Calcium oxalate deposition observed in vitamin B6 deficient kidney was completely prevented when fed along with vitamin E or methionine. However the hyperoxaluria and hypercalciuria persisted even after feeding with vitamin E or methionine.

Restoration of antioxidants in liver by methionine feeding in experimental rat urolithiasis.

The effect of methionine or citrate on antioxidant defense system has been studied in urolithic rat. Liver weight and its protein concentration did not change in the rats fed with calculi producing diet (CPD) when compared to normal diet fed rats. Feeding rats along with citrate (c-CPD) or methionine (m-CPD) improved their body weight gain. Liver microsomes and mitochondria fractions of CPD and c-CPD fed groups showed increased susceptibility for lipid peroxidation in presence of ascorbate and t-butyl hydroperoxide when compared to either control or m-CPD fed groups. Increased superoxide dismutase and xanthine oxidase activities, decreased catalase, glutathione peroxidase and glucose-6-phosphate dehydrogenase activities, decreased concentrations of reduced glutathione, total thiols, ascorbic acid and vitamin-E and increased formation of hydroxyl radical, hydroperoxides and diene conjugates were observed in the liver of both CPD fed group as well as c-CPD fed group. Except SOD and xanthine oxidase, all other parameters were normalized in m-CPD fed group. This suggested that feeding methionine reduced the susceptibility for lipid peroxidation by restoration of the level of free radical scavengers.

Effect of citrate feeding on free radical induced changes in experimental urolithiasis.

Feeding calculi producing diet (CPD) to rats for 4 weeks produced calcium oxaltate stones. Supplementation of sodium citrate to CPD (c-CPD) prevented stone formation. Except oxalate, the excretion of calcium, phosphorus and magnesium was restored to normal in c-CPD fed rats. The CPD fed rats exhibited increase in glycolic acid oxidase (GAO) and lactate dehydrogenase (LDH) activities and only GAO activity was partially restored in c-CPD fed rats. Kidney sub-cellular fractions of calculi producing diet (CPD) fed rats showed increased susceptibility for lipid peroxidation in presence of promotors. Antioxidant enzyme activities of superoxide dismutase (SOD), catalase and glutathione peroxidase and antioxidant concentrations of reduced glutathione, total thiols, ascorbic acid and vitamin E were significantly decreased while the xanthine oxidase activity, and concentrations of hydroxyl radical, diene conjugates and hydroperoxides were significantly increased in CPD fed rats. The susceptibility to lipid peroxidation, activities of antioxidant enzymes, and the concentration of antioxidants were not normalized by feeding citrate.

Glucose-6-phosphate dehydrogenase deficiency and malaria--a study on north Madras population.

Blood samples from 381 healthy individuals and 236 malaria patients residing in North Madras were studied for glucose-6-phosphate dehydrogenase deficiency. The incidence of this deficiency in this area was found to be 10.05%. Partially deficient healthy females showed a protective trend against malarial infection with the Chi-squared test approaching statistical significance.

Increased plasma lipidperoxidation in vitamin B-6 deficient rats.

Lipidperoxidation in plasma of rats fed with vitamin B-6 deficient diet for a period of 12 weeks was studied with pair-fed controls. Plasma pyridoxal 5'-phosphate, alanine amino transferase and aspartate amino transferase, the markers of vitamin B-6 status, were significantly low in vitamin B-6 deficient rats. Plasma malondialdehyde level, conjugated dienes and lipofuscin like pigments were increased in vitamin B-6 deficiency. Increased levels of plasma lipids, calcium, iron and copper were observed in vitamin B-6 deficiency. Plasma susceptibility to lipidperoxidation was maximal in vitamin B-6 deficiency, upon stimulation by the promotors, Fe2+, Fe3+, Cu2+, ascorbate, t-butyl hydroperoxide and hydrogen peroxide.

Lipids changes in rat tissues in experimental urolithiasis.

Total lipids, free and ester cholesterol, triglyceride and phospholipids were determined in plasma, liver, kidney and intestine in control and calculi producing diet (CPD) fed rats. Cholesterol, phospholipids and triglycerides were increased in plasma while they were decreased in all the three tissues of CPD fed rats, compared to that of control. Distribution studies of phospholipids in the tissues of treated rats showed marked decrease in the concentration of the major lipids, i.e., PC, PE, PI and SPH. However, significant increase in absolute concentration as well as percent distribution of phosphatidic acid in kidney of treated rats was observed.

Induction of lipid peroxidation in calcium oxalate stone formation.

The function of lipid peroxidation and the anti-peroxidative enzymes of rat liver and kidney were investigated under hyperoxaluric and stone forming conditions. The experimental animals showed higher malondialdehyde content in liver and kidney than that of control. A significant increase in malondialdehyde release was observed in the experimental liver or kidney when incubated with either ferrous sulphate or hydrogen peroxide compared to that of control liver or kidney. Superoxide dismutase activity was not affected in the hyperoxaluric rats while there was a moderate increase in the stone forming rats when compared to control. Highly significant decrease in catalase activity was observed in both conditions in liver and kidney compared to control.

Increased lipid peroxidation in the erythrocytes of kidney stone formers.

The level of lipid peroxidation was significantly increased in erythrocytes and erythrocyte membrane in patients with stone disease. Increased activities of catalase and acetylcholinesterase in the erythrocyte membrane were observed, while hemolysate displayed no significant change in superoxide dismutase activity. The amount of phospholipids in the RBC membrane of patients was significantly increased. Peroxidation was stimulated by oxalate in vitro and was further enhanced in the presence of ferrous ion. The changes in lipid peroxidation and antioxidant enzymes are suggestive of chemical alteration of the RBC membrane during urolithiasis.

Induction of lipid peroxidation by oxalate in experimental rat urolithiasis.

The function of lipid peroxidation and the antiperoxidative enzymes of rat liver and kidney were studied in stone formation induced by intraperitoneal administration of sodium oxalate (7 mg/100 g body weight). The animals sacrificed 3 and 12 h after administration of sodium oxalate had higher level of malondialdehyde in liver and kidney than control animals. A significantly pronounced release of malondialdehyde was observed in treated liver and kidney homogenates when incubated with either ferrous sulphate or hydrogen peroxide compared to control liver and kidney. Superoxide dismutase activity was increased only in liver and not in kidney in treated animals compared to the control. A highly significant decrease in catalase activity was observed in both liver and kidney of treated animals.