Indian Journal of Biochemistry & Biophysics
CODEN: IJBBBQ ISSN:
0301-1208
|
VOLUME 46 |
NUMBER 1 |
FEBRUARY 2009 |
Special
Issue
Free Radicals and Antioxidants in Human Health
CONTENTS
Minireviews
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Free
Radical Induced Oxidative Damage to DNA: Relation to Brain Aging and |
9 |
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Endogenous
and Induced Oxidative Stress in Multi-cellular Tumour Spheroids: |
16 |
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Is Mycophenolate
more than just an Immunosuppressant? – An Overview |
25 |
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Future
Perspectives of Nutrigenomics Foods: Benefits vs. Risks |
31 |
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Papers
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HBx
protein modulates PI3K/Akt pathway to overcome genotoxic stress-induced |
37 |
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|
|
|
|
|
|
Protective effect of
liposomal formulation of tuftsin (a naturally occurring tetrapeptide) against
cyclophosphamide-induced genotoxicity and oxidative stress in mice |
45 |
|
K Arif,
A Ejaj, A Maroof, A K Azmat, C Arun,
|
|
|
|
|
|
Induction of
oxidative stress by restraint stress and corticosterone treatments in rats |
53 |
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|
|
|
|
|
|
Curcumin arrests
endometriosis by downregulation of matrix metalloproteinase-9 activity
|
59 |
|
|
|
|
|
|
|
Evaluation
of the chronic complications of diabetes in a high fructose diet in rats |
66 |
|
|
|
|
|
|
|
Role of bilirubin
as antioxidant in neonatal jaundice and effect of ethanolic extract of sweet lime peel on
experimentally induced jaundice in rat |
73 |
|
|
|
|
|
|
|
Effect
of Bacillus subtilis PB6, a natural probiotic on colon mucosal
inflammation and |
79 |
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R Selvam, P Maheswari*, P Kavitha, M
Ravichandran, B Sas &
C
|
|
|
|
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Functionality
of drug efflux pumps in antimonial resistant Leishmania donovani field
|
86 |
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G Mandal, A Sarkar, P Saha, N Singh,
|
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Stress modulating antioxidant effect of Nardostachys jatamansi |
93 |
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N Lyle, D Bhattacharyya*, T
K Sur, S Munshi, S Paul,
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Antidiabetic
and antioxidant potential of ethanolic extract of Butea monosperma
leaves in
|
99 |
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Embryo protective
effect of pomegranate (Punica granatum L.) fruit extract in adriamycin-induced
oxidative stress |
106 |
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Antioxidant activity of commonly consumed cereals, millets, pulses and legumes in
|
112 |
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Time-dependent
effects of ethanol on blood oxidative stress parameters and cytokines |
116 |
|
S K Das*, S Varadhan, G Gupta, S
Mukherjee, L Dhanya, D
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Short communications
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Antioxidant potential of the
root of Vetiveria zizanioides (L.) Nash |
122 |
|
S
Luqman*, R Kumar, S Kaushik, S Srivastava, M P Darokar & S P
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Improved method of total antioxidant assay |
126 |
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Alterations
in plasma nitric oxide during aging in humans |
130 |
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Author Index |
133 |
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Keyword Index |
134 |
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——————
*Author
for correspondence
Foreword
Free Radical Research is an interdisciplinary area of
research involving basic scientists, clinicians, therapists, pharmacists,
biotechnologists and food technologists. The Society for Free Radical
Research (SFRR) promotes research on free radicals and antioxidants with
particular reference to medical and industrial importance for the benefit of
mankind. SFRR-India held its third satellite meeting at the All India Institute
of Medical Sciences (AIIMS),
Prof. Rao’s meticulous job of pooling the papers, get
them peer-reviewed and revised, and the editorial team’s painstaking dedicated
efforts in bringing out this fine special issue on the theme “Free Radicals
and Antioxidants in Human Health” are indeed adorable. The contents cover a
wide range of topics in the general area of Free Radical Research with reviews
on oxidative DNA damage in relation to aging and neural disorders, oxidative
stress in tumour spheroids, which are highly relevant ex vivo model
systems for cancer; mycophenolate for immunosuppression and other beneficial
effects; and pros and cons of nutrigenomics foods. The regular papers pertains
to genotoxic stress; restraint stress and corticosterone treatment as a
mediator of oxidative stress; mechanisms responsible for curcumin induced
inhibition of endometriosis; high fructose diet-induced diabetes model and its
implications; tuftsin and cyclophosphamide induced genotoxicity; beneficial
effect of bilirubin and sweet lime peel on neonatal jaundice; probiotic and
inflammatory bowel disease; drug efflux and leishmaniasis; stress modulation by
the Nardostachys jatamansi; antidiabetic and antioxidant effect of Butea
monosperma; pomegranate and adriamycin toxicity; and antioxidant effect of
cereals, millets, pulses and legumes. Short communications included antioxidant
potential of Vetiveria zizaniodes; improved method for antioxidant
assay; and alterations in nitric oxide levels during aging.
The above manuscripts cover a wide variety of highly
relevant topics in Free Radical Research, and I am sure they will be of great
interest to the readers. I acknowledge all the authors for their contributions,
reviewers for cooperation, besides the editorial team of IJBB for hardwork and
the unstinted support of The Director, NISCAIR without whom publication of this
flawless issue would not have been possible.
Dr. T.P.A. Devasagayam
Vice-President, SFRR-India & President, SFRR-Asia
February 24, 2009
Indian Journal of
Biochemistry & Biophysics
Vol. 46, February 2009, pp
9-15
MINIREVIEWS
Free Radical Induced Oxidative damage to DNA: Relation to Brain Aging and Neurological Disorders
Kalluri Subba Rao*
Center for Biotechnology,
Jawaharlal Nehru Technological University (JNTU), Kukatpally,
Received 12 June 2008; revised 10 November 2008
Free radicals are produced in cells by cellular metabolism and by exogenous agents. These species react with biomolecules in cells and one of the important targets is DNA. This kind of damage, often referred to as oxidative DNA damage, has consequences in various organs and particularly in brain, in view of its high metabolic activity and oxygen consumption. The consequences include mutagenesis of various kinds ranging from simple oxidation of bases to large deletions through single and double strand breaks. In brain, because of its post-mitotic nature, oxidative damage to DNA is seen more often at the level of bases. A major route for repairing oxidative damage to bases is base excision repair (BER). It is increasingly becoming apparent that defects in repairing oxidative DNA damage can lead to a number of neurological disorders like Alzheimer and Parkinson. Our recent studies have clearly demonstrated that BER is highly compromised in brain cells with increasing age and this could well be one of the major causative factors for normal aging and the associated deteriorating mental conditions, including certain neurological abnormalities.
Keywords:
Free radical, Oxidative damage, DNA, Brain aging, Neurological disorders, Base excision repair (BER), Nucleotide
excision repair (NER)
E-mail: ksrsl@yahoo.com
Indian Journal of
Biochemistry & Biophysics
Vol. 46, February 2009, pp.
16-24
Endogenous and Induced Oxidative Stress in Multi-cellular Tumour Spheroids: Implications for Improving Tumour Therapy
Divya Khaitan and B
Division of Radiation Biosciences, Institute of Nuclear
Medicine and Allied Sciences,
Brig. S K Mazumdar Marg, Delhi 110054, India
Received 17 July 2008; revised 9 January 2009
The endogenous oxidative stress in tumours is determined by the status of mitochondrial, metabolic, oxygen (hypoxia) and inherent enzymatic as well as non-enzymatic antioxidant defense systems, which influence tumour growth and respond to anticancer therapeutics. Induced oxidative stress is one of the important determinants of the outcome of treatment with certain chemotherapeutic drugs and ionizing radiation. The mild to moderate levels of reactive oxygen species (ROS) have often been found to trigger prosurvival responses, thereby contributing to the resistance against therapy. The higher levels of ROS stimulate multiple death pathways viz. typical and atypical apoptosis, necrosis etc, thereby enhancing the therapeutic efficiency. Therefore, approaches employing therapeutic agents that generate ROS efficiently in the tumour cells and enhance the antioxidant defense system in the normal cells could significantly enhance the therapeutic gain. Multi-cellular tumour spheroids (MCTS) offer an excellent in vitro system that mimics endogenous oxidative stress often observed in tumours, arising due to a number of factors (gradients of oxygen and nutrients, altered intercellular interaction and tumour necrosis factor), besides antioxidant defense systems similar to tumours in vivo. More importantly, MCTS resemble tumours in vivo with reference to the induced oxidative stress related responses, particularly following combinations of certain chemotherapeutic drugs and metabolic inhibitors and differs significantly from the responses in monolayer cultures. Therefore, MCTS appear to be excellent in vitro models, ideally suited for developing novel therapies that are based on the generation of oxidative stress in tumours. The present review provides a modest account on the utility of MCTS in understanding the role of oxidative stress in treatment-induced responses of tumours for designing therapies and therapeutics.
Keywords:
Multi-cellular spheroids, Oxidative stress, Radiotherapy, Chemotherapy
E-mail: bsd@inmas.org
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 25-30
Is Mycophenolate more than just
an Immunosuppressant? –
An Overview
Abishek Iyer and Lindsay Brown*
School of Biomedical Sciences, The
Received 10 June 2008; revised 22 January 2009
The development of immunosuppressant compounds, such as cyclosporine and tacrolimus was crucial to the success of transplant surgery and for treatment of autoimmune diseases. However, immunosuppressant therapy may increase the concentrations of reactive oxygen species (ROS), inducing oxidative damage such as an increased vascular damage. The major source of ROS in the vascular endothelial cells is NADPH oxidase. The subunit structure and function of this enzyme complex in vascular cells differs from that in phagocytic leucocytes. The enzyme subunits Nox1, Nox2 and Nox4 are only found in vascular cells. The GTP-dependent protein subunit Rac 1 needs to be activated for this enzyme to function. Inhibiting this protein subunit should reduce NADPH oxidase-induced oxidative stress. In the cardiovascular system, oxidative stress is observed as hypertension, hypertrophy, fibrosis, conduction abnormalities and endothelial dysfunction, as well as cardiac allograft vasculopathy in transplant patients. In contrast to cyclosporine and tacrolimus, the immunosuppressant mycophenolate inhibits the Rac 1 subunit thus inhibiting NADPH oxidase in the vasculature. This may reduce oxidative stress, prevent the development of cardiac allograft vasculopathy, decrease the deterioration of vascular function and improve cardiovascular function chronically in transplant patients. This overview discusses whether this anti-oxidant immunosuppressive property could translate into a more general protective role for mycophenolate in the prevention of cardiovascular disease.
Keywords: Mycophenolate,
Immunosuppressants, Oxidative stress, NADPH oxidase, Cardiovascular disease
E-mail:
l.brown@uq.edu.au
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 31-36
Future Perspectives of Nutrigenomics Foods: Benefits vs. Risks
Dilip Ghosh*
Smart Foods Centre,
Received 20 September 2008; revised 10 November 2008
Nutrigenomics, defined as the application of high-throughput genomics tools in nutrition research is now past its incubation phase. The poorly understood associations of diet and disease prevention in particular will likely be the single most important catalyst to its accelerated and continued growth. Whether the goal of matching foods to individual genotypes to improve the health of those individuals can be attained, and personalised nutrigenomic foods enter the world's food markets, depends on numerous hurdles being overcome: some scientific in nature, some technical and others related to consumer, market or ethical issues. Public adoption of new technologies is an important determinant for their success. Many of the drivers behind the trend in personalisation of food are now known, particularly ethical, legal, and social issues (ELSI) are the major drivers. Future development in the field of nutrigenomics undoubtedly will place its seemingly huge potential in better perspective. From the scientific responsibility point of view, one hopes that the new perspectives to be gained and progress to be made in this field will be so managed as to take the public at large on board, if we are to avoid another nutrition education disaster of the genetically modified organism type and dimension.
Keywords: Nutrigenomics, Nutrigenetics, Nutrigenomics foods, Functional foods, Nutrition, Consumers, Ethical-legal-social issues, Regulation,
Gene-diet interactions,
Diet, Omics
E-mail:
dilip.ghosh@bigpond.com
PAPERS
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 37-44
HBx protein modulates PI3K/Akt pathway to overcome genotoxic stress-induced destabilization of cyclin D1 and arrest of cell cycle
A Mukherji, V C
Janbandhu and V Kumar*
Virology Group, International Centre for Genetic Engineering
and Biotechnology, Aruna Asaf Ali Marg, New Delhi 110067, India
Received 23 April 2008; revised 15 January 2009
Growth arrest represents an innate barrier to carcinogenesis. DNA damage and replicational stress are known to induce growth arrest and apoptotic death to avert genomic instability and consequently carcinogenesis. In this study, working on the genotoxic stress induced by hydroxyurea and methylmethanesulfone, we observed a growth arrest at G1/S-phase that was mediated by destabilization of cyclin D1. The growth arrest was independent of the stability of cdc25A and preceded transcriptional up-regulation of p21waf1. Cyclin D1 destabilization involved its phosphorylation by GSK-3b at threonine-286, since overexpression of the kinase-dead mutant of GSK-3b or cyclin D1T286A mutant conferred stability to cyclin D1. Further, overexpression of cyclin D1T286A also helped in bypassing G1/S phase growth arrest. We also observed a rapid inactivation of Akt/PKB kinase in the presence of hydroxyurea. Enforced expression of the constitutively active Akt or viral oncoprotein HBx (Hepatitis B virus X protein) was sufficient to overcome growth arrest, independent of ATR signaling and stabilized cyclin D1. Thus, the present work not only establishes cyclin D1 to be a novel mediator of genotoxic stress signaling, but also explains how a deregulated mitogenic signaling or a viral oncoprotein can help bypass growth arrest.
Keywords:
Cell cycle, Cyclin D1, Genotoxic stress, HBx, Hydroxyurea,
Methylmethanesulfone, PKB/Akt
E-mail: vijay@icgeb.res.in
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp 45-52
Protective effect of liposomal formulation of tuftsin (a naturally occurring tetrapeptide) against cyclophosphamide-induced genotoxicity and oxidative stress in mice
K Arif, A Ejaj, A Maroof, A K Azmat, C Arun,
Interdisciplinary Biotechnology Unit,
+
Received 02 August 2008; revised 16 January 2009
Tuftsin, a naturally occurring tetrapeptide with a sequence Thr-Lys-Pro-Arg was evaluated for its in vivo protective effect against cyclophosphamide-induced genotoxicity and oxidative stress in Swiss albino mice. The anticancer drug cyclophosphamide (CP) was administered intra-peritonially to induce mutagenic effect. The drug treatment caused significant increase in chromosomal aberrations, formation of micronucleated polychromatic erythrocytes (MNPCE’s), as well as oxidative stress and decrease in lipid peroxidation in liver of the animals. The pretreatment with tuftsin abolished such effects in dose-dependent manner and also increased mitotic index in the experimental animals. Results of the present study validated chemo-preventive properties of tuftsin against CP-induced chromosomal mutations and cellular injury of liver by oxidative stress.
Keywords: Genotoxicity, Cyclophosphamide, Tuftsin, Liposome,
Oxidative stress
E-mail: owais_lakhnawi@yahoo.com
Indian Journal of Biochemistry & Biophysics
Vol. 46, February 2009, pp. 53-58
Induction of oxidative stress by restraint stress and
corticosterone
treatments in rats
Ayesha Zafir and Naheed Banu*
Department of
Biochemistry, Faculty of Life Sciences,
Received 16 July 2008; revised 21 January 2009
Chronic exposure to psychological stress in humans and restraint stress in experimental animals results in increased oxidative stress and resultant tissue damage. To study the contribution of stress hormones towards stress-induced oxidative processes in the brain, we investigated the response of important free-radical scavenging enzymes toward chronic administration of two doses of corticosterone (low dose: 10 mg/kg/day, high dose: 40 mg/kg/day) in rodents. After a 21-day experimental period, a significant decline in both superoxide dismutase and catalase was observed in both stressed and stress hormone-treated animals. The brain levels of glutathione as well as the activities of glutathione-S-transferase and glutathione reductase were also significantly decreased, while lipid peroxidation levels were significantly increased in comparison to controls. A direct pro-oxidant effect of stress hormones in the brain during physical and psychological stress was observed, indicating important implications for oxidative stress as a major pathological mechanism during chronic stress and a consequent target option for anti-stress therapeutic interventions.
Keywords: Oxidative stress, Restraint stress,
Antioxidant enzymes, GSH, Lipid peroxidation, Corticosterone, Stress hormone
Email: naheedbanu7@yahoo.com
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 59-65
Curcumin arrests endometriosis by downregulation of
matrix metalloproteinase-9 activity
Snehasikta Swarnakar* and Sumit
Paul
Department of Physiology, Drug Development Diagnostics
and Biotechnology Division,
Indian Institute of Chemical Biology,
4, Raja S.C. Mullick Road, Jadavpur, Kolkata 700032, India
Received 06 October 2008; revised 17 January 2009
Curcumin, a polyphenol derived from turmeric (Curcuma longa) possesses diverse pharmacological properties including antioxidant, anti-inflammatory and antiproliferative activities. Endometriosis is a gyneocological disorder characterized by growth of endometrial tissues outside uterus that involves aberrant matrix remodeling. In this study the effect of curcumin was studied on surgically developed endometriosis in mice. Endometriosis with varying severity was developed in mice by peritoneal implantation of uterine fragments. The changes in matrix metalloproteinase (MMP)-9 and tissue inhibitor of metalloprotease (TIMP)-1 were investigated in endometriotic tissues following curcumin pre- and post-treatment. Results showed that MMP-9 activity increased gradually in endometriotic tissues with severity and curcumin treatment reversed the MMP-9 activity near to control value. Curcumin administered either post- or pre-endometriosis arrested endometriosis in a dose-dependent manner. It inhibited both MMP-9 activity and its expression at the level of secretion, during regression of endometriotic lesion. In addition, the attenuated activity of MMP-9 was associated with decreased expression of tumor necrosis factor-α (TNF-α) during healing, suggesting the anti-inflammatory property of curcumin. Moreover, curcumin pretreatment prevented lipid peroxidation and protein oxidation in endometriotic tissues. We reported here for the first time the anti-endometriotic property of curcumin via MMP-9 dependent pathway that may lead to new therapeutic intervention.
Keywords: Curcumin, Endometriosis, Matrix metalloproteinase, Tissue
inhibitor of metalloproteinase, Tumor necrosis factor-a, Extracellular matrix
Email: snehasiktas@hotmail.com
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp 66-72
Evaluation of the chronic complications of diabetes in a high fructose diet in rats
Jatin Patel, Abishek Iyer and Lindsay Brown*
School of Biomedical Sciences, The
Received 12 June 2008;
revised 19 January 2009
The increasing prevalence of type 2 diabetes is associated with increasing health costs, especially for the treatment of cardiovascular disease. The development of new treatment modalities requires animal models that mimic the range of pathophysiological changes seen in diabetic humans. Dietary fructose intake has been linked to the increase in insulin resistance as part of the metabolic syndrome; fructose-fed rats develop type 2 diabetes. This study has characterized the cardiovascular changes in young adult male Wistar rats fed a 61% fructose diet for 16 weeks. Our results extend the reported changes of hypertension, lipid abnormalities, impaired glucose tolerance and impaired oxidative defense to include ventricular dilatation with hypertrophy and decreased contractile function, together with increased inflammatory cell infiltration into the ventricular myocardium, resulting in excessive collagen deposition and an increased stiffness of the left ventricle. However, endothelial dysfunction, tactile allodynia as a symptom of peripheral neuropathy and retinopathy are not present in these rats, in contrast to the streptozotocin-induced model of type 1 diabetes. Thus, fructose feeding mimics many, but not all, of the symptoms of type 2 diabetes in humans.
Keywords: Type 2 diabetes, High fructose diet
E-mail: l.brown@uq.edu.au
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 73-78
Role of bilirubin as antioxidant in neonatal jaundice and
effect of ethanolic extract of sweet lime peel on experimentally induced
jaundice in rat
N Nag, S Halder, R Chaudhuri, S Adhikary† and
*Department of Biochemistry,
Kolkata 700 019,
†Department of Radiation and Photochemistry, Bhabha
Atomic Research Centre,
Received 23 July 2008; revised 16 January 2009
Bilirubin above a threshold level is toxic to human system and is excreted in urinary and through gastrointestinal tract. The role of bilirubin as antioxidant is debatable. This paper aims at elucidating the role of bilirubin as an antioxidant in neonatal jaundice patients. It is observed that bilirubin up to 6 mg/dl in blood acts as an antioxidant and above 12.5 mg/dl is strongly prooxidant. Phototherapy is the accepted therapeutic management of neonatal jaundice and has been shown to enhance the oxidative stress. Approaches have been taken to formulate a herbal medication which will reduce bilirubin level in the neonates without inducing additional damages. The ethanolic extract of sweet lime peel, administered orally at a dose of 72 µg is found to reduce the oxidative stress in erythrocytes of phenylhydrazine-induced jaundiced rats treated with phototherapy.
Keywords: Bilirubin,
Antioxidant, Neonatal jaundice, Oxidative stress, Phototherapy, Sweet lime peel
Email: smbioc@gmail.com
Indian
Journal of Biochemistry & Biophysics
Vol. 46, February 2009, pp.
79-85
Effect of Bacillus
subtilis PB6, a natural probiotic on colon
mucosal inflammation and plasma cytokines levels in
inflammatory bowel disease
R Selvam1, P Maheswari3*, P Kavitha4,
M Ravichandran2, Benedikt Sas6 and C
1Preclinical Development, 2Research and
Development, Kemin Industries South Asia (P) Ltd, Plot # K3, 11th
Cross Street, SIPCOT Industrial complex, Gummidipoondi, 601 201, Tamil Nadu,
India
3Preclinical Development, 4Clinical
Development, 5Research and Development, Kemin Pharma, Kemin
Industries South Asia (P) Ltd, Pharma, The
Trapezium, # 39, Second Floor, Nelson Manickam Road,
Chennai 600 029,Tamil Nadu, India
6Kemin
Pharma bvba, Atealaan 4H, B-2200 Herentals,
Received 04 August 2008;
revised 15 January 2009
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 × 108 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-α, IL-1β, IL-6 and IFN-γ) are also significantly lowered (p<0.05) and anti-inflammatory cytokine (IL-10 and TGF-β) 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.
Keywords:
Inflammatory bowel disease, Bacillus
subtilis PB6, TNBS-induced
colitis, Wistar rats, Cytokines
E-mail: maheshwari.p@kemin.com
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp 86-92
Functionality of drug efflux pumps in antimonial resistant Leishmania donovani field isolates
Goutam Mandal1, Avijit Sarkar1, Piu Saha1, Neeloo Singh2, and Shyam Sundar3 and Mitali Chatterjee1*
1Department
of Pharmacology, Institute of Post Graduate Medical Education and Research,
Kolkata, 244B Acharya JC Bose Road, Kolkata 700 020 India
2Drug
Target Discovery & Development Division, Central Drug Research Institute,
3Dept.
of Medicine,
Received 15 May 2008; revised 10 January 2009
The recent upsurge of antimony (Sb) resistance is a major impediment to successful chemotherapy of visceral leishmaniasis (VL). Mechanisms involved in antimony resistance have demonstrated an upregulation of drug efflux pumps; however, the biological role drug efflux pumps in clinical isolates remains to be substantiated. Thus, in this study, the functionality of drug efflux pumps was measured in promastigotes and axenic amastigotes isolated from VL patients, who were either Sb-sensitive (AG83, 2001 and MC9) or resistant (NS2, 41 and GE1) using rhodamine123 as a substrate for multidrug resistant (MDR) pumps and calcein as a substrate for multidrug resistance-associated proteins (MRP) respectively; their specificity was confirmed using established blockers. Sb-resistant (Sb-R) isolates accumulated higher amounts of R123, as compared to Sb-sensitive (Sb-S) isolates. Verapamil, a MDR inhibitor failed to alter R123 accumulation, suggesting absence of classical MDR activity. In Sb-R isolates, both promastigotes and axenic amastigotes accumulated significantly lower amounts of calcein than Sb-S isolates and probenecid, an established pan MRP blocker, marginally increased calcein accumulation. Depletion of ATP dramatically increased calcein accumulation primarily in Sb-R isolates, indicating existence of a MRP-like pump, which was more active in Sb-R isolates. In conclusion, our data suggested that overfunctioning of a MRP-like pump contributed towards generation of Sb-R phenotype in L. donovani field isolates.
Keywords: Antimony resistance, Axenic amastigotes, Drug efflux pumps, Multidrug
resistance protein (MRP), Multidrug resistant (MDR),
Promastigotes, Visceral leishmaniasis
E-mail: ilatim@vsnl.net
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 93-98
Stress modulating antioxidant effect of Nardostachys jatamansi
Nazmun Lyle1, Dipankar
Bhattacharyya1*, Tapas K Sur1, Santanu
Munshi2, Suhrita Paul3, Suparna
Chatterjee1 and
1Department
of Pharmacology,
244-B,
2Department
of Pharmacology,
3Department
of Pharmacology,
4Department of Physiology,
University College of Science and Technology,
University of Calcutta,
Received 25 March 2008 ; revised 29 October 2008
The rhizomes of Nardostachys jatamansi, the plant commonly known as Jatamansi have been described in Ayurveda for their soothing and sedative action on the central nervous system. In the present study, the anti-stress effect of hydro-ethanolic extract (70%) of N. jatamansi (NJE) was evaluated in reference to its antioxidant property. Wistar rats were divided into four groups: naïve, stressed, and T-200 and T-500 stressed with oral pre-treatment of NJE 200 and 500 mg/kg, respectively. Restraint of rats in metallic chambers for 4 h at 4°C was followed by sacrifice and assessment of stress-induced alterations in biochemical parameters, incidence and severity of ulcers. Lipid peroxidation (LPO) and NO levels in stomach and LPO, NO levels and catalase activity in brain, plasma corticosterone level and adrenal ascorbic acid were measured. In vitro antioxidant activity of NJE was studied by measuring the free radical scavenging activity. NJE showed potent antioxidant activity and significantly reversed the stress-induced elevation of LPO and NO levels and decrease in catalase activity in the brain. It inhibited the incidence of gastric ulcerations and reversed the alterations in biochemical parameters/markers of stress-induced gastric ulceration. NJE also significantly altered stress-induced increase in adrenal and spleen weights and decrease in level of ascorbic acid in adrenal gland. Elevation of plasma corticosterone level was negated dose- dependently. The findings suggest that the NJE possesses significant anti-stress activity, which may be due to its antioxidant activity.
Keywords:
Adaptogen, Corticosterone, Lipid peroxidation, Nardostachys jatamansi,
Restraint stress, ROS, Ulcer
E-mail: lyle.nazmun@gmail.com;
lyle_n@rediffmail.com
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 99-105
Antidiabetic and antioxidant potential of
ethanolic extract of
Butea monosperma leaves in
alloxan-induced diabetic mice
Nidhi Sharma* and Veena Garg
Department of Bioscience and Biotechnology,
Received 05 July; revised 25 November 2008
The possible protective effect of ethanolic extract of B. monosperma leaves (BMEE) on diabetes and diabetes-induced oxidative stress was evaluated in alloxan (ALXN)-induced diabetic male adult mice. Experimental animals were divided into three groups viz., I, II, and III. Diabetes mellitus (DM) was induced in groups II and III mice by a single intraperitoneal injection of alloxan (150 mg/kg body wt). Group I (control mice) received an equal volume of normal saline. Group III mice were further treated with BMEE (300 mg/kg body wt, p.o.) for a period of 45 days. Body weight and fasting blood glucose (FBG) levels were measured at periodic intervals during the test period. At the end of treatment period, blood was collected by cardiac puncture under mild ether anesthesia and serum was isolated to analyze its lipid profile i.e. serum total cholesterol (TC), triglyceride (TG), high density lipoprotein (HDL), low density lipoprotein (LDL) and very low density lipoprotein (VLDL). The homogenates of hepatic, pancreatic and renal tissues were also analyzed for both enzymatic and non-enzymatic antioxidants, such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), reduced glutathione (GSH), thiobarbituric acid reactive substances (TBARS) and total protein (TP). Alloxan injection resulted in a significantly (P<0.05) increased concentration of FBG level. Besides, the levels of enzymatic and non-enzymatic antioxidants were decreased and TBARS level increased significantly (P<0.05) in hepatic, pancreatic and renal tissues. Also, serum TC, TG, LDL and VLDL-cholesterol level elevated significantly (P<0.05), whereas HDL-cholesterol reduced significantly (P<0.05) in group II (alloxan-treated diabetic control). The FBG level decreased significantly (P<0.05) after 45 days treatment of BMEE from 172 to 117.143 mg/dl, as compared to normal control (79.286 mg/dl). The activities of antioxidant enzymes (CAT and GSH-Px) and GSH level in hepatic, pancreatic and renal tissues also increased significantly (P<0.05) in BMEE-treated mice, but the activity of SOD was not improved significantly. BMEE treatment also reduced the TBARS levels and lowered serum lipid profile significantly (P<0.05). The findings of the present study indicated significant hypoglycemic and anti-oxidant activity in B. monosperma leaves, thus lends credence to its folklore use in the management and/or control of type-2 DM.
Keywords: Butea monosperma
leaves, Diabetes mellitus, Oxidative stress, Alloxan, Antidiabetic, Antioxidant
potential
E-mail: nidhisharma2006@gmail.com;
sharma_nidhi18@yahoo. co.in
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 106-111
Embryo
protective effect of pomegranate (Punica
granatum L.)
fruit extract in adriamycin-induced oxidative stress
R
Department of Biochemistry,
Received 21 June 2008; revised 31 December 2008
The possible protective role of pomegranate (Punica
granatum L.)
fruit extract which has shown antioxidant capacity higher than that of red wine
and green tea was evaluated against adriamycin-induced oxidative stress in
chick embryos. Adriamycin (ADR), an anthracycline broad spectrum of
chemotherapeutic drug is used for the treatment of variety of cancers; however,
its prolonged use is limited by an irreversible, dose-dependant and progressive
cardiomyopathy, hepatotoxicity and general toxicity to other organs in human
beings, due to oxidative stress. The morphological changes (malformation of
different organs), changes in body weight, volume of amniotic fluid (AF) and
biochemical parameters of AF were studied after 24 and 48 h of incubation by
comparing ADR alone and pomegranate fruit extract pretreated groups with their
respective controls of 12 days old chick embryos. ADR alone at a dose of 70 mg/egg showed a significant dose versus time-
dependent reduction in body weight, volume of AF. A dose-related increase in
embryo gross morphological deformities and significant changes in the levels of
biochemical parameters in AF were observed in ADR-treated group. These changes
were significantly ameliorated to normal by pre-administration of pomegranate
fruit extract at a dose of 200 µg/egg. Thus,
the present study demonstrated the embryo protective nature of pomegranate
fruit extract against ADR-induced oxidative stress.
Keywords: Adriamycin, Amniotic fluid, Biochemical parameters,
Chick embryo
E-mail: prparthasarathy85@yahoo.co.in
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 112-115
Antioxidant
activity of commonly consumed cereals, millets, pulses and legumes in
D Sreeramulu*, C Vijaya Kumar Reddy and M Raghunath
Endocrinology
and Metabolism Division, National Institute of Nutrition,
Received 16 October 2008; revised 16 January 2009
Plant foods are
important due to their antioxidant activity (AOA) attributed to the phenolics
which are known to protect organisms against harmful effects of oxygen
radicals. However, information on antioxidant activity of Indian plant foods is
scanty. Therefore, the present study evaluated the AOA of cereals, millets,
pulses and legumes, commonly consumed in
Keywords: Antioxidant activity, Ferric reducing antioxidant
power, Diphenyl-1-picryl hydrazyl, Total
phenolic content, Cereals, Millets, Pulses, Legumes
Email:
dandesr@yahoo.com
Indian
Journal of Biochemistry & Biophysics
Vol. 46, February 2009, pp. 116-121
Time-dependent effects of ethanol on blood oxidative stress parameters and cytokines
Subir Kumar Das†*, Sowmya Varadhan, Geetanjali Gupta#,
Sukhes Mukherjee, L Dhanya,
D N Rao# and D M Vasudevan
Department of Biochemistry,
Amrita Institute of Medical Sciences, Elamakkara P.O. Cochin 682 026, Kerala
†
#All India Institute of Medical Sciences, Ansari
Nagar,
Received 15 June 2008; revised 06 January 2009
Alcohol
consumption is implicated in the genesis of a spectrum of liver abnormalities,
which are associated with a number of factors. In the present study,
time-dependent effects of ethanol on cytokines (TNF-a, IL-2, IL-4, IL-10, IFN-g, VEGF-A and
TGF-b1) in serum, and blood oxidative stress parameters
such as reduced glutathione content, TBARS level and activities of
GPx, GR, GST, catalase and SOD in 8-10 weeks-old male BALB/c mice have been
investigated. Ethanol administered @1.6 g/kg body wt/day significantly
increased the activities of liver marker enzymes AST, ALT and ALP. Serum
nitrite levels and haemolysate TBARS level also increased, while total
antioxidant status in serum and GSH content in whole blood hemolysate decreased
from 4th week onwards of exposure. Inspite of the increased serum
nitrite level and GST activity in the haemolysate, albumin level in serum, GPx
and GR activities in haemolysate decreased after 12 weeks of exposure. Chronic
ethanol treatment did not show any effect on IL-2, but IL-4 level was reduced
and other cytokines such as IL-10, TNF-a, IFN-g, TGF-b1 and VEGF-A levels were increased
significantly after 12 weeks. The study indicates a relationship between free
radical generation and immune response, and suggests that ethanol-induced liver
damage is associated with oxidative stress and immunological alterations in a
time-dependent manner.
Keywords: Cytokines, Ethanol, Interleukin, Liver, Oxidative
stress
E-mail: drsubirkdas@yahoo.co.in; drsubirkdas@gmail.com
SHORT COMMUNICATIONS
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 122-125
Antioxidant potential of the root of
Vetiveria zizanioides (L.)
Nash#
Suaib Luqman*, Ritesh Kumar, Shubhangi Kaushik, Suchita Srivastava, Mahendra P Darokar and Suman P S Khanuja
Genetic Resources and Biotechnology Division, Central
Institute of Medicinal and Aromatic Plants (Council of Scientific and Industrial
Research), P.O. CIMAP,
Received 17 July 2008; revised 24 December 2008
Vetiveria zizanioides, an aromatic plant commonly known as vetiver has been used for various ailments. The essential oil of vetiver root has been shown to possess antioxidant activity. However, antioxidant potential of spent root extract has not been reported. Hence, in the present study, ferric reducing, free radical scavenging and antioxidant activity of two genotypes namely KS1 and gulabi of V. zizanioides L. Nash root were investigated using in vitro assays — the ferric reducing antioxidant power (FRAP), 1,1-diphenyl-2-picrylhydrazyl (DPPH), total phenolic content (TPC), total antioxidant capacity (TAC) and reducing power (RP). KS1 genotype showed higher FRAP values, DPPH inhibition, TPC and RP potential compared to gulabi and the antioxidant activity increased with the concentration of the extract (10-1000 µg/mL). A significant protective effect of cv KS1 (100 µg/mL) extract was also observed in reduced glutathione (GSH) and malondialdehyde (MDA) concentrations of erythrocytes subjected to oxidative stress by tert-butyl hydroperoxide (t-BHP) and hydrogen peroxide (H2O2). The cv KS1 showed better antioxidant activity, compared to cv gulabi indicating the possibility of exploring the presence of different phytoconstituents in the two varieties.
Keywords: Vetiveria zizanioides, Antioxidant, Erythrocytes, Glutathione,
Malondialdehyde
E-mail: s.luqman@cimap.res.in
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp. 126-129
Improved method of total antioxidant assay
Ruby Guptaa, Mukta Sharmaa, Ramakrishnan
Lakshmya*, Dorairaj Prabhakaranb
and K Srinath Reddyc
aDepartment of Cardiac Biochemistry, cDepartment
of Cardiology,
All India Institute of Medical Sciences, Ansari Nagar,
New Delhi 110029
bCenter for Chronic Disease Control, Safdarjung
Development Area,
Received 06 October 2008; revised 20 January 2009
Commercially available analytical kits for the estimation of total antioxidant status are expensive and time-consuming. Most of the commercially available kits for total antioxidants estimation are based on the principle of suppression of ABTS radical cation formation by antioxidant in the serum sample. The method requires stringent assay conditions, like exact incubation time and the temperature (37°C) of the reaction and on an average not more than 40 samples can be analyzed on a day. We have adapted the assay to a microplate, thereby allowing more number of samples to be analyzed per day. Further, the reagent volume required is one fourth than that for the original procedure thereby cutting cost. Thirty samples were analyzed by original method on spectrophotometer and our adapted microplate assay. The values of total antioxidant obtained by the two methods correlated well. Thus, total antioxidant can be estimated reliably using the microplate method.
Keywords: Total antioxidant, ABTS, Microplate, Trolox,
Coefficient of variation
E-mail: lakshmy_ram@yahoo.com; lakshmyram@gmail.com
Indian
Journal of Biochemistry & Biophysics
Vol.
46, February 2009, pp 130-132
Alterations in plasma nitric oxide during aging in humans
Pawan Kumar Maurya and Syed Ibrahim Rizvi*
Department of Biochemistry,
Received 16 July 2008; revised 20 January 2009
Nitric oxide (NO) is relatively harmless, but along with superoxide radical becomes precursor of many toxic species, such as peroxy and hydroxyl radicals, hydrogen peroxide, and peroxynitrite. In the present study, we determined plasma NO as a function of human age and correlated NO levels with total antioxidant capacity of the plasma. Results showed significant increase in NO level as a function of human age and plasma NO level positively correlated with total antioxidant potential. Increased NO may contribute to the development of oxidative stress during aging.
Keywords: Nitric oxide, Aging, Human, Oxidative stress
E-mail: rzv@rediffmail.com
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