Mugineic acid, active ingredient of wheat grass: an oral novel hexadentate iron chelator in iron overloaded diseases.

Iron chelation therapies are required for the treatment of iron overloaded patients; nonetheless, their side effects are also well known. We have evaluated iron-chelating activity of wheat grass extract (WHE) and its purified compound, mugineic acid in murine model with phenylhydrazine (PHZ) and dextran induced acute and chronic iron overload conditions. PHZ and dextran treatment induced acute and chronic iron overload condition in mice, respectively, as indicated by increased serum and tissue iron in both cases. Iron overload was also accompanied with haemosiderosis in tissues (liver and spleen). These PHZ and dextran -: treated mice were orally treated with either crude WHE or purified mugineic acid. The efficacy of mugineic acid and WHE was compared with the potent oral iron chelator ICL670 (Exjade). PHZ and dextran treatment followed by oral administration of WHE or mugineic acid significantly checked the rise of serum/plasma levels of iron as well as tissue iron and also, haemosiderosis in tissues. The results are highly comparable with known iron chelator ICL670. WHE and purified mugineic acid, both seem to have significant prospect to be the cheap, non-toxic, hexadentate and oral therapeutic agents to prevent or alleviate toxic iron overload in patients.

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.

Governance of HIV/AIDS: implications for health sector response.

This paper reviews the essence of effective governance and importance of a multi-sectoral approach in generating health systems response to HIV/AIDS. This comprehensive approach highlights the importance of integrating reproductive sexual health programs and HIV prevention services, including peer education, life skills, and Voluntary Counseling and Testing (VCT), for Prevention of Mother-to-Child Transmission (PMTCT) and reaching out to People Living with HIV/AIDS (PLHA). Research implications for governance of health systems response to HIV/AIDS, integrated youth health policies and high-level political commitment, are emphasized by strategic implications for HIV/AIDS control and followed by a policy thrust on health systems as a strategic plan to achieve sustainability in the fight against HIV/AIDS.

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

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.

Study of rogi kalyan samitis in strengthening health systems under national rural health mission, district pune, maharashtra.

BACKGROUND: With objective of health systems strengthening, as visualized under National Rural Health Mission (NRHM); one key strategic intervention is up-gradation of health service delivery facilities so as to provide sustainable quality care with accountability and people's participation, which required the development of a proper management structure called Rogi Kalyan Samitis (RKS). It is the State's attempt to make health everyone's business by de-mystifying health-care delivery at district and sub-district levels with reference to facility based health-care delivery by encouraging citizen's participation in management bodies. OBJECTIVE: The study was an attempt to define 'functional Health Systems' with a focus on strategic issues concerning RKS operations. MATERIALS AND METHODS: A mixed-method, multi-site, collective case study approach was adopted. In-depth interviews of key-stakeholders were conducted. Qualitative data were analyzed thematically and coded inductively. RESULTS: RKS is yet to bring out quality component to the health services being provided through facilities. This can be attributed to structural and managerial weakness in the system; however, certainly NRHM has been consistent in creating a road-map for benefitting local community and their participation through RKS. CONCLUSION: The progress of the RKS can further be enhanced by giving due priority to critical areas. Furthermore, the results emphasize an urgent need for devising strategies and actions to overcome significant systemic constraints as highlighted in the present study.

Distortion of ß-globin chain of hemoglobin alters the pathway of erythrocytic glucose metabolism through band 3 protein.

BACKGROUND AND AIMS: Band 3 is a transmembrane protein of erythrocytes and its cytosolic part regulates glucose metabolism to proceed along the pentose phosphate pathway (PPP) or glycolytic pathway by binding with the central cavity of the ß chain of deoxygenated hemoglobin and with some glycolytic enzymes competitively. In ß thalassemia major and hemoglobin (Hb)Eß thalassemia, ß chain is either absent or distorted and glucose metabolism may be disturbed. We estimated adenosine triphosphate (ATP), glucose 6-phosphate dehydrogenase (G6PD) and aldolase activity in oxygenated and deoxygenated state of erythrocytes of ß major, HbEß thalassemia and normal controls to understand the hypothesis that major glucose metabolism within the erythrocytes of these diseases may proceed through the PPP. METHODS: Fifty of each group of patients and 52 normal controls were included. Patients' blood was collected 1 month prior to blood transfusion. G6PD and aldolase were estimated using a commercial kit. ATP was measured by spectrophotometric method. RESULTS: Significantly low levels of erythrocytic ATP and higher levels of G6PD were found in two groups of patients. Aldolase level in deoxygenated hemolysate was significantly higher than oxygenated hemolysate in normal controls but no significant change was noticed in both types of thalassemic patients. CONCLUSIONS: In ß thalassemia major and HbEß thalassemia due to distortion of ß chain, binding of deoxygenated hemoglobin with band 3 is inhibited and thus traffic control of glycolytic pathway is disturbed and shifted towards PPP.

A novel biflavonyloxymethane from Pongamia pinnata and its radical quenching activity.

The root bark of Pongamia pinnata Pierre (syn P. glabra Vent.) has afforded a new biflavonyloxymethane, pongabiflavone, along with a known furanoflavone, 3-methoxy-(7, 8, 2", 3") furanoflavone. The structure of this new compound was elucidated from extensive spectral studies, including 2D-NMR spectroscopic experiments. The antioxidant, radical quenching activity- superoxide and nitric oxide quenching activities of both pongabiflavone and previously isolated karanjabiflavone have been evaluated which can be a key to cure Psoriasis.

Non-haem iron-mediated oxidative stress in haemoglobin E beta-thalassaemia.

INTRODUCTION: Haemoglobin (Hb) E beta-thalassaemia is a common thalassaemic disorder in Southeast Asia and is very common in the eastern and north-eastern parts of India. The disease cause rapid erythrocyte destruction due to the free radical mediated injury but factors for the oxidative injury are not clearly known. We investigated the free reactive iron (non-haem) mediated insult in Hb E beta-thalassaemia. MATERIALS AND METHODS: Thirty Hb E beta-thalassaemic patients (age range, 3 to 15 years) who had undergone blood transfusion at least 1 month prior to sampling and 32 normal healthy individuals (age range, 18 to 30 years) were included in this study. We estimated the ferrozine detected intracellular erythrocytic free reactive iron (nonhaem iron), reduced glutathione (GSH), glutathione reductase activity, cellular damage marker serum thiobarbituric acid reacting substances (TBARS) and also serum ferritin using standard methods. RESULTS: We found that the erythrocytic free reactive iron was significantly higher (P <0.001) in Hb E beta patients and was about 30% more than in controls. The elevated level of erythrocytic non-haem iron was associated with a high level of serum TBARS which was about 86% higher in patients than in controls. The serum ferritin level was also significantly higher (P <0.001) compared to controls. The erythrocytic reduced glutathione level was significantly lower (P <0.001) at about 65% less in the patients' group and the erythrocytic glutathione reductase enzyme was also found to be significantly lower (P <0.001) in Hb E beta-thalassaemia. CONCLUSIONS: We concluded that a significantly elevated level of erythrocytic free reactive iron and lipid peroxidation end product was associated with low erythrocytic GSH level. This reflects non-haem iron mediated cellular damage in Hb E beta-thalassaemia.

Elevated level of carbonyl compounds correlates with insulin resistance in type 2 diabetes.

INTRODUCTION: Recent periodicals direct that reactive carbonyl compounds are formed due to existing oxidative stress in type 2 diabetes mellitus, which further nonenzymatically react with proteins and lipids to form irreversible advanced glycation end products (AGE) and advanced lipoxidation end products (ALE). In type 2 diabetes mellitus, insulin resistance plays a pivotal role in hyperglycaemia. In this study, we tried to fi nd the relation between insulin resistance and carbonyl stress. MATERIALS AND METHODS: Forty-seven patients of type 2 diabetes mellitus (age 51 ± 5.06 years) were selected and fasting plasma glucose, serum insulin, total carbonyl compounds, HbA1c, thiobarbituric acid reacting substances (TBARS) and Trolox equivalent antioxidant capacity (TEAC) were estimated using standard protocols. Homeostatic model assessement of insulin resistance (HOMA-IR) was evaluated from fasting plasma glucose and serum insulin levels. RESULTS: We found highly significant correlations of carbonyl compounds with HOMA-IR, fasting plasma glucose and glycated haemoglobin (HbA1c). Correlations of lipid peroxidation end product, TBARS were not so significant. CONCLUSION: Findings from this study indicate that the level of carbonyl compounds can be a biomarker of insulin resistance in type 2 diabetes mellitus.

Antidiarrhoeal activity of carbazole alkaloids from Murraya koenigii Spreng (Rutaceae) seeds.

The bioassay guided fractionation of the n-hexane extract of the seeds of Murraya koenigii Spreng (Rutaceae) resulted in the isolation of three bioactive carbazole alkaloids, kurryam (I), koenimbine (II) and koenine (III). The structures of the compounds were confirmed from their (1)H-, (13)C-, and 2D-NMR spectral data. Of the three compounds (I) and (II) exhibited significant inhibitory activity against castor oil-induced diarrhoea and PGE(2)-induced enteropooling in rats. The compounds also produced a significant reduction in gastrointestinal motility in the charcoal meal test in Wistar rats.

A new chalcone from Pongamia pinnata and its antioxidant properties.

The root bark of Pongamia pinnata Pierre [syn P. glabra (family: Fabaceae)] afforded a new chalcone (karanjapin) and two known flavonoids, a pyranoflavonoid (karanjachromene) and a furanoflavonoid (karanjin) The structure of karanjapin has been established from extensive 2D NMR spectral studies as beta,2'-dihydroxy-a,4'-dimethoxy-3,4-methylenedioxychalcone. Karanjapin and karanjachromene were found to possess significant antioxidant activity. This may play an important role in the pathogenesis of several diseases.

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.

Effect of glycation of hemoglobin on its interaction with trifluoperazine.

Trifluoperazine (TFZ), a phenothiazine drug, penetrates into human erythrocytes and releases oxygen by interaction with hemoglobin. TFZ-induced oxygen release from hyperglycemic erythrocytes isolated from diabetic patients is considerably less compared to that from the cells of normoglycemic individuals. In diabetes mellitus, hemoglobin is significantly glycated by glucose. Non-glycated hemoglobin, HbA0 and its major glycated analog, HbA1c have been separated from the blood samples of diabetic patients. TFZ releases considerable amount of oxygen from HbA0, but very little from HbA1c. Spectrofluorimetric studies reveal that TFZ forms excited state complexes with both HbA0 and HbAlc. Titration of HbA0 with TFZ in a spectrophotometric study exhibits two isosbestic points. Similar experiment with HbAlc causes gradual loss of the Soret peak without appearance of any isosbestic point indicating a possibility of heme loss during interaction, which is also supported by gel filtration experiment and SDS-PAGE experiment followed by heme staining. The results suggest that drug action on hemoglobin is influenced by glycation-induced structural modification of the protein.

Effect of nonenzymatic glycation on functional and structural properties of hemoglobin.

HbA(1c), the major glycated hemoglobin increases proportionately with blood glucose concentration in diabetes mellitus. H(2)O(2) promotes more iron release from HbA(1c) than that from nonglycated hemoglobin, HbA(0). This free iron, acting as a Fenton reagent, might produce free radicals and degrade cell constituents. Here we demonstrate that in the presence of H(2)O(2), HbA(1c) degrades DNA and protein more efficiently than HbA(0). Formation of carbonyl content, an index of oxidative stress, is higher by HbA(1c). Compared to HbA(0), HbA(1c) is more rapidly autooxidized. Besides these functional changes, glycation also causes structural modifications of hemoglobin. This is demonstrated by reduced alpha-helix content, more surface accessible hydrophobic tryptophan residues, increased thermolability and weaker heme-globin linkage in HbA(1c) than in its nonglycated analog. The glycation-induced structural modification of hemoglobin may be associated with its functional modification leading to oxidative stress in diabetic patients.