Manoj Kar

Role of free reactive iron in psoriasis.

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In vitro Studies of Iron Chelation Activity of Purified Active Ingredients Extracted from Triticum aestivum Linn. (Wheat Grass)

Objective: Seven to eight days germinated plants of Triticum aestivum (wheat grass) are a rich source of vitamin A, C, calcium, magnesium, phosphorus, potassium, sodium, sulphur, cobalt, zinc and protein. Traditionally the aqueous extract of T. aestivum was reported to be used as a health tonic in folk and ayurvedic medicine. We previously reported that aqueous extract of T. aestivum was found to reduce the blood transfusion requirement in iron overloaded Thalassemia and Myelodysplastic syndrome patients. Our objective was to extract and purify active ingredients from wheat grass and study their mode of action in stabilizing hemoglobin level in those patients. Design and Method: Active ingredients of wheat grass were extracted and purified by cation exchange column chromatography followed by High Performance Liquid Chromatography. In vitro experiments with phenylhydrazine treated red blood cell hemolysate were carried out before and after treatment of purified fraction of T. aestivum to study iron chelating activity. Result: Purified fraction of T. aestivum treated red blood cell showed significant inhibition

Suppression of NF-κB p65 nuclear translocation and tumor necrosis factor-α by Pongamia pinnata seed extract in adjuvant-induced arthritis

Abstract Pongamia pinnata is a plant known for its therapeutic usage in Indian traditional medicine. Despite the controversy regarding toxic flavonoid and erucic acid content, the seed of this plant is consumed in tribal medicine and its oil is used in Ayurveda to treat psoriasis and arthritis. This study explored the potential anti-arthritic effects of a P. pinnata seed (hexane) extract (PSE) at non-lethal doses in an adjuvant-induced arthritic rat model; possible mechanisms of any observed effects were also explored. After establishing the lethal doses arising from oral exposure to the extract, the material was administered per os daily at two doses (0.3 g/kg/day; 0.5 g/kg/day) to arthritic rats. Other rats received indomethacin or vehicle (control). Treatments were performed for a total of 14 days. One day after the final exposure, the rats were euthanized to permit harvest of various cells, blood, and tissues for analyses. Paw diameter and tissue myeloperoxidase activity in the paws were evaluated as indices for edema and neutrophil infiltration into the tissue. The severity of arthritis in the experimental rats was assessed via measures of urinary hydroxyproline (HP) and glucosamine, and of serum pro-inflammatory TNFα and anti-inflammatory IL-10. The extent of NF-κB p65 nuclear translocation in peritoneal macrophages harvested from naïve rats and then treated in vitro was also assessed. The results indicated that exposure to PSE significantly decreased paw diameter, tissue myeloperoxidase level, and levels of urinary HP and glucosamine, as well as of serum TNFα and IL-10 in adjuvant-injected (arthritic) rats. In vitro PSE treatment also resulted in a marked inhibition of NF-κB p65 nuclear translocation in primary cultures of peritoneal macrophages. Thus, PSE appears to be able to prevent experimental arthritis, in part, by helping to maintain the balance between pro- and anti-inflammatory cytokines and by inhibiting NF-κB activation.

Methylglyoxal increase in uremia with special reference to snakebite-mediated acute renal failure

BACKGROUND Advanced glycation and lipoxidation endproducts (AGEs and ALEs) due to oxidative and carbonyl stress are involved in pathogenesis of several diseases including uremia. Methylglyoxal, a dicarbonyl compound is a metabolic hazard and potent glycating agent in the body, which is an important precursor of AGE and ALE. Methylglyoxal has been reported to be increased in uremia, but there is no report of MG status in snake venom mediated acute renal failure cases (SARF). We investigated the carbonyl and oxidative stress as well as the methylglyoxal concentration in SARF where renal clearance is rapidly shut down. METHODS We studied serum carbonyl compounds, methylglyoxal, total antioxidant status, GSH and cellular damage marker thiobarbituric acid reacting substances (TBARS) and intracellular erythrocytic GSH concentration following standard methods of 45 SARF and 56 normoglycemic chronic renal failure cases (CRF) and compared with 81 normal controls. RESULT Methylglyoxal concentration has been found to be significantly increased in SARF associated with decreased concentration of serum as well as erythrocytic GSH and other antioxidant markers , in comparison with CRF and normal control. The cellular damage (TBARS concentration), is also found increased in SARF. CONCLUSION MG increase as well as accumulation due to GSH depletion may play a pivotal role in their rapid pathophysiological complicacies in SARF.

Quantifying morphological alteration of RBC population from light scattering data

Studies of RBC morphological alterations, despite their potential clinical and experimental application, are compromised due to lack of simple and rapid techniques. As a complementary approach toward quantitative microscopy, we have reconstituted morphological information from light scattering data obtained from flow cytometer. Normal and poikilocytic agent treated samples were analyzed by microscopy and respective morphological index (MI) was calculated from the morphology based scores assigned to RBC. The samples were simultaneously analyzed by flowcytometer and the scatter data were obtained. Accordingly, the best correlated parameters of both forward scatter and side scatter were chosen to formulate a suitable regression model with MI as response. Flow cytometry data was also verified with another instrument (BD FACS Verse) and the equation obtained was validated with separate set of samples. The multivariate regression analysis yields a quadratic model with MI as response (R2 = 0.96, p <  0.001). The flow cytometric data from both instruments were in good agreement (Intra class correlation ∼0.9, p <  0.001). The model was found to simulate the sample MI with high accuracy (R2 = 0.97, p <  0.001). This proposed method was verified to be simple, rapid, quantitative and cost effective for the measurement of morphological alteration of RBC.