Indian Journal of Experimental Biology

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VOLUME 47

NUMBER 2

FEBRUARY 2009

CODEN: IJEB (A6) 47(2) 77-150 (2009)

ISSN: 0019-5189

CONTENTS

Papers

 

   
Celecoxib administration exhibits tissue specific effect on 3H-benzo(a)pyrene-DNA adduct formation in cigarette smoke inhaling mice 83
Ashwani Koul, Lalita Tanwar & Neha Arora  
   
In vivo microdialysis studies of striatal level of neurotransmitters after haloperidol and chlorpromazine administration
91
Shrinivas K Kulkarni, Mahendra Bishnoi & Kanwaljit Chopra  
   
Influence of w-6/w-3 rich dietary oils on lipid profile and antioxidant enzymes in normal and stressed rats 98
M K Benson & Kshama Devi  
   
Beneficial effects of modified egg on oxidative stress in F1- generation of metabolic syndrome-X induced Wistar rat 104
Satish Kumar Taneja & Kshetrimatyum Birla Singh  
   
Anti-inflammatory activity of flower extract of Calendula officinalis Linn. and its possible mechanism of action
  113
Korengath Chandran Preethi, Girija Kuttan & Ramadasan Kuttan  
   
Antimicrobial activity of flavanoid sulphates and other fractions of Argyreia speciosa (Burm.f) Boj
  121
P V Habbu, K M Mahadevan , R A Shastry & H Manjunatha  
   
Administration of aqueous extract of Desmodium gangeticum (L) root protects rat heart against ischemic reperfusion injury induced oxidative stress 129
Gino A Kurian & Jose Paddikkala  
   
Antagonistic effect of polyamines on ABA-induced suppression of mitosis in
Allium cepa L.
136
Arpana Mahajan & Shashi Sharma  
   
High frequency plantlet regeneration from rhizomatous buds in Mantisia spathulata Schult. and Manitisia wengeri Fischer and analysis of genetic uniformity using RAPD markers 140
Sudipta Shekhar D Bhowmik, Suman Kumaria, S R Rao & Pramod Tandon  
   
Notes  
   
Effect of thyroxineon plasminogen activator and inhibitor activity in rat 147
P Bubber, N Bubber  & D D Bansal  

 

Author Index

Arora Neha

83

 

 

Bansal D D

147

Benson M K

98

Bhowmik Sudipta Shekhar D

140

Bishnoi Mahendra

91

Bubber  N

147

Bubber P

147

 

 

Chopra Kanwaljit

91

 

 

Devi Kshama

98

 

 

Habbu P V

121

 

 

Koul Ashwani

83

Kulkarni Shrinivas K

91

Kumaria Suman

140

Kurian Gino A

129

Kuttan Girija

113

Kuttan Ramadasan

113

 

 

Mahadevan K M

121

Mahajan Arpana

136

Manjunatha H

121

 

 

Paddikkala Jose

129

Preethi Korengath Chandran

113

 

 

Rao S R

140

 

 

Sharma Shashi

136

Shastry R A

121

Singh Kshetrimatyum Birla

104

 

 

Tandon Pramod

140

Taneja Satish Kumar

104

Tanwar Lalita

83

 

 

 

 

Keyword Index

ABA

136

Allium cepa

136

Antifungal

121

Anti-inflammatory agents

113

Antimicrobial

121

Antioxidants

129

Antitubercular

121

Argyreia speciosa

121

Atherogenic index

98

Atypical antipsychotics

91

 

 

Calendula officinalis

113

Carcinogenesis

83

Carcinogen-DNA adduct

83

Catalase

136

Celecoxib

83

Cigarette smoke

83

Cu

104

Cyclooxygenase-2

113

 

 

Desmodium gangeticum

129

Diabetes mellitus

104

Fibrinolytic parameters

147

Flavanoid sulphates

121

 

 

Hepatic

83

Homeostatic balance

147

 

 

In vivo microdialysis

91

Inhalation

83

Lipid peroxidation

98

 

 

Manitisia wengeri

140

Mantisia spathulata

140

Mg

104

Mitotic Index

136

Mn

104

Modified egg

104

Mustard oil

98

Myocardial ischemia

129

Myocardial ischemic
 reperfusion injury

 

129

 

 

Neurotransmitters

91

Omega-3 fatty acid

104

Oxidative stress

104

 

 

Plant regeneration

140

Proinflammatory cytokines

113

PUFA

98

Pulmonary

83

Putrescine

136

 

 

RAPD analysis

140

Renal function

147

 

 

Spermidine

136

Spermine

136

Sunflower oil

98

 

 

Tardive dyskinesia

91

Thyroxine

147

 

 

Vitamin- E

104

 

 

Zn

104

 

 

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 83-90

 

 

Papers

 

Celecoxib administration exhibits tissue specific effect on
3H-benzo(a)pyrene-DNA adduct formation in cigarette smoke inhaling mice

Ashwani Koul*, Lalita Tanwar & Neha Arora

Department of Biophysics, Basic Medical Sciences Block, Panjab University, Chandigarh 160 014, India

Received 2 April 2008; revised 1 December 2008

In the present study, cigarette smoke (CS) exposure significantly enhanced 3H-B(a)P-DNA adduct formation in both the pulmonary and hepatic tissues. Mice co-treated with CS and celecoxib (a specific COX-2 inhibitor) exhibited a significant decrease in hepatic carcinogen-DNA adduct formation in comparison to the smoke exposed group, however the lungs of the co-treated animals exhibited a significant increase in carcinogen-DNA adduct formation when compared to the control group and smoke exposed group. CS exposure enhanced the activity of carcinogen activation enzymes in both the tissues and decreased the activity of carcinogen detoxification enzymes in the hepatic tissue only, when compared to the control group. Celecoxib administration to CS inhaling mice modulated the carcinogen biotransformation considerably when compared to the CS exposed group. Celecoxib administration to CS inhaling mice produced a low index of carcinogenesis in the hepatic tissue but increased the index of carcinogenesis in the pulmonary tissue. These observations seem to be critical and tissue specific when related to carcinogenesis.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 91-97

 

 

In vivo microdialysis studies of striatal level of neurotransmitters after haloperidol and chlorpromazine administration

Shrinivas K Kulkarni*Ş, Mahendra Bishnoi & Kanwaljit ChopraŞ

Centre with Potential for Excellence in Biomedical Sciences (CPEBS), Panjab University, Chandigarh, 160 014, India

ŞPharmacology Division, University Institute of Pharmaceutical Sciences, Panjab University, Chandigarh, 160 014, India

Received 4 September, 2008; revised 15 December 2008

Therapeutic success of atypical antipsychotics has focused the attention on the role of receptor systems other than dopaminergic system in the pathophysiology of neuroleptics-associated acute (Parkinson’s like syndrome) and chronic (tardive dyskinesia) extrapyramidal side effects. This study was planned to investigate changes in striatal levels of norepinephrine, dopamine and serotonin after acute and chronic administration of classical neuroleptics (haloperidol and chlorpromazine). These changes were correlated with behavioural alterations in rats. In vivo microdialysis with HPLC/ECD system revealed that there was a marked decrease in striatal neurotransmitter contents (NE, DA and 5-HT), which was also correlated with severe cataleptic response in rats after acute administration of haloperidol (2 mg/kg) and chlorpromazine (20 mg/kg). Chronic administration of haloperidol (1 mg/kg for 21 days) and chlorpromazine (5 mg/kg for 21 days) resulted in time dependent increase in orofacial hyperkinetic movements. The microdialysis studies also showed a significant decrease in the striatal levels of all the neurotransmitters. The results provide evidence for the involvement of striatal adrenergic and serotonergic systems, besides dopaminergic system in neuroleptic-induced acute and chronic extrapyramidal symptoms.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 98-103

 

 

Influence of w-6/w-3 rich dietary oils on lipid profile and antioxidant enzymes in normal and stressed rats

M K Benson & Kshama Devi*

Department of Pharmacology, Al-Ameen College of Pharmacy, Bangalore, 560 027, India

Received 12 June 2008; revised 17 December 2008

To evaluate the influence of w-6/w-3 poly unsaturated fatty acid (PUFA) containing oils on lipid profile and endogenous antioxidant enzymes in normal and stressed (immobilization) rats, 28 day old male Wistar rats were fed for 45 days with fat enriched special diet (10% fat) prepared with sunflower oil (SO) ľ w-6 rich, mustard oil (MO) ľ w-3 rich and groundnut oil ľ control respectively. SO treated normal rats have significantly reduced total cholesterol, high density lipoprotein – cholesterol (HDL-C) and catalase thereby significantly increased the atherogenic index (AI) and lipid peroxidation (LPO). However, treatment with MO increased superoxide dismutase; decreased LPO significantly. Under stress conditions AI and LPO were significantly high with SO and significantly less with MO. In addition, SO decreased HDL-C whereas MO decreased non-HDL-C significantly. Results suggest a protective role against AI and LPO in normal and stress conditions in MO. The quantity of w-3 fatty acids in dietary oil may play a crucial role in the body against atherogenicity. The findings signify that not just PUFA, but type of PUFA present in dietary oil used is important.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 104-112

 

 

Beneficial effects of modified egg* on oxidative stress in F1- generation of metabolic syndrome-X induced Wistar rat

Satish Kumar Taneja & Kshetrimatyum Birla Singh

Department of Zoology, Panjab University, Chandigarh 160 014, India

Received 23 August 2007; revised 5 December 2008

Congenital malformations of neonates are one of the adverse effects of diabetic pregnancy which can be prevented by supplementation of vitamin E and C. The survived neonates usually are at higher risk to diabetes, hypertension, dyslipidemia and cardiovascular diseases that may possibly be prevented through antioxidants administration. In view of this information, the efficacy of modified poultry egg enriched with optimum minerals, vitamin E and omega-3 fatty acids was studied on F1-generation, which were made to survive by feeding them this modified egg to diabetic mothers of Wistar rats. The survived F1-generation displayed hyperglycemia, dyslipidemia and hypertension like their parents, evaluated after three months of the experiment. Their mineral status revealed a higher Zn and lower Cu, Mg and Mn levels in liver and kidney. Their lipid peroxidation products were however higher and the enzyme activities of superoxide dismutase, catalase, glutathione-S-transferase, glutathione reductase, glutathione (reduced) and glucose -6 phosphate dehydrogenase were significantly lower. In the other group of F1-generation, fed modified egg mixed diet, a significant reduction in the blood pressure, serum glucose, serum lipid profile, and the lipid peroxidation products, and a significant increase in the activities of enzymes per se with reversal of Zn, Cu, Mg and Mn levels closer to the control group were recorded. The data suggest that the modified egg can ameliorate the oxidative stress in F1- generation of diabetic rats by improving the mineral status in their body.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 113-120

 

 

Anti-inflammatory activity of flower extract of Calendula officinalis Linn. and its possible mechanism of action

Korengath Chandran Preethia, Girija Kuttanb & Ramadasan Kuttana*

Department of  aBiochemistry and bImmunology Amala Cancer Research Centre, Amala Nagar, Thrissur 680 555, India

Received 7 March 2008; revised 18 December 2008

Calendula officinalis flower extract possessed significant anti-inflammatory activity against carrageenan and dextran-induced acute paw edema. Oral administration of 250 and 500 mg/kg body weight Calendula extract produced significant inhibition (50.6 and 65.9% respectively) in paw edema of animals induced by carrageenan and 41.9 and 42.4% respectively with inflammation produced by dextran. In chronic anti-inflammatory model using formalin, administration of 250 and 500 mg/kg body weight Calendula extract produced an inhibition of 32.9 and 62.3% respectively compared to controls. TNF-α production by macrophage culture treated with lipopolysaccharide (LPS) was found to be significantly inhibited by Calendula extract. Moreover, increased levels of proinflammatory cytokines IL-1β, IL-6, TNF-α and IFN-γ and acute phase protein, C- reactive protein (CRP) in mice produced by LPS injection were inhibited significantly by the extract. LPS induced cyclooxygenase-2 (Cox-2) levels in mice spleen were also found to be inhibited by extract treatment. The results showed that potent anti-inflammatory response of C. officinalis extract may be mediated by the inhibition of proinflammatory cytokines and Cox-2 and subsequent prostaglandin synthesis.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 121-128

 

 

Antimicrobial activity of flavanoid sulphates and other fractions of Argyreia speciosa (Burm.f) Boj.

P V Habbu1*, K M Mahadevan2 , R A Shastry1 & H Manjunatha3

1Postgraduate Department, Division of Pharmacognosy, SET’s College of Pharmacy, Dharwad 580 002, India

2Postgraduate Department, School of Chemical Sciences and Research, Kuvempu University, Shankaraghatta, Shimoga, India

3Department of PG Studies and Research in Biotechnology and Bioinformatics Kuvempu University, Shankaraghatta 577 45, India

Received 22 July 2008; revised 12 December 2008

Antimicrobial activity of flavanoid sulphates and different fractions of A. speciosa root was studied against bacteria, fungi and Mycobacterium tuberculosis H37 Rv sensitive strain by in vitro and in vivo assays. Flavanoid sulphates such as quercetin 3’7 di-O methyl 3- sulphate and kaempferol 7-O methyl 3-sulphate were isolated from the n-butanol fraction of 80% methanolic extract of the plant. The structures of the isolated flavanoids were confirmed by spectral studies. Ethyl acetate (EAAS) fraction and flavanoid sulphates inhibited the growth of M. tuberculosis Rv sensitive strain at MIC values 50 and 25 mg/ml, respectively. Ethanolic fraction (EtAS) showed significant inhibition of gram positive organism with a MIC of 31.25 µg/ml. More inhibition was observed with a less MIC (2 µg/ml) for flavanoid sulphates against Klebsiella pneumoniae, a gram negative organism and it is almost comparable with the standards. Interestingly, chloroform fraction alone exhibited significant antifungal activity with a MIC of 100 µg/ml. A synergistic effect between flavanoids sulphates and commercially available antitubercular drugs was observed with FIC index of 0.443±0.245, 0.487±0.247 for isoniazid and 0.468±0.333, 0.417±0.345 for rifampicin, whereas EAAS fraction showed partial synergistic effect. A synergistic effect was observed for EAAS fraction and flavanoids sulphates with FIC index < 0.5 with antibiotics. Hemolysis assay on RBCs suggested that EAAS and flavanoids sulphates exhibited least cellular toxicity to erythrocytes as compared to chloramphenicol. In vivo studies in mice infected with K. pneumoniae demonstrated that on day 10 post treatment of different fractions and isolated compounds of A. speciosa, about 60% of the animals treated with EAAS, 70% of animals treated with flavanoids sulphates and 40% of animals treated with EtAS were survived.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 129-135

 

 

Administration of aqueous extract of Desmodium gangeticum (L) root protects rat heart against ischemic reperfusion injury induced oxidative stress

Gino A Kurian*

MMM Academy of Medical Sciences, Institute of Cardiovascular Diseases, Madras Medical Mission,
4A Jayalalithanagar, Mogappair, Chennai 600 037, India

and

Jose Paddikkala

Department of Plant Biotechnology, Amala Cancer Research Center, Amalanagar, Trichur 680 555, India

Received 25 July 2007; revised 11 November 2008

Myocardial reperfusion is believed to be associated with free radical injury. The present study evaluates the effect of aqueous extract of D. gangeticum (DG) on lipid peroxides and antioxidants in ischemic reperfused (IR) Wistar albino male rats. Significant elevation in lipid peroxide products (thiobarbituric acid reactive substances) and decreased activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase) were observed in the rat hearts during ischemia reperfusion phase. Pre treatment of rats with aqueous extract of DG orally for 30 days showed significantly improved preservation of antioxidant enzymes and subsequent reduction in lipid peroxidation. But 2,3,5 triphenyl tetrazolium chloride (TTC) stained rat heart did not show much significant antioxidant enzyme activities and lipid peroxidation. On the other hand, TTC unstained rat heart showed significant improvement in the antioxidant activities indicating cardio protective effect of aqueous extract of DG in myocardium affected by ischemia reperfusion insult. The administration of DG to normal rats did not have any significant effect on any of the parameter studied. These results indicate that DG improves the antioxidant capacity of heart and attenuate the degree of lipid peroxidation after IR.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 136-139

 

 

Antagonistic effect of polyamines on ABA-induced suppression of mitosis
in Allium cepa L.

Arpana Mahajan & Shashi Sharma*

Department of Biosciences, Himachal Pradesh University, Shimla 171 005, India

Received 17 September 2008; revised 27 November 2008

Effect of abscisic acid (ABA) and polyamines (PAs) [putrescine (Put), spermidine (Spd) and spermine (Spm)] on mitosis in root tips of A. cepa was studied. Treatment with ABA (0.1 to 100 µM) for 24 hr suppressed the mitosis, measured as mitotic index (MI), in a concentration-dependent manner with approx. 50% suppression at 10 µM of ABA. Treatment with different PAs (1 to 100 µM) had differential mitosis suppression effect. Spm was most inhibitory followed by Spd and Put, respectively. The higher concentrations of PAs (1 mM Put; 0.1 and 1 mM Spd or Spm) caused cell distortion. Remarkably, a 24 hr pretreatment of root tips with PAs prior to ABA (100 µM) treatment resulted in a general concentration-dependent reversal of ABA-induced suppression of MI. Catalase (CAT) activity in the root tips, an indicator of redox metabolism, increased due to ABA treatment in a concentration-dependent manner, remained unaltered in response to Put and declined due to Spd and Spm (≥0.1 mM). However, all PAs, irrespective of their individual effects, generally antagonized the ABA-dependent increase in CAT activity. Data indicate the possibility of ABA-PA interaction in the regulation of mitosis.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 140-146

 

 

High frequency plantlet regeneration from rhizomatous buds in Mantisia spathulata Schult. and Mantisia wengeri Fischer and analysis of genetic uniformity using RAPD markers

Sudipta Shekhar D Bhowmik, Suman Kumaria*, S R Rao** & Pramod Tandon

Plant Biotechnology Laboratory, Department of Botany,

Centre for Advanced Studies, North Eastern Hill University, Shillong 793 022, India

Received 25 August 2008; revised 21 November 2008

A protocol has been devised for enhanced in vitro regeneration of critically endangered Mantisia spathulata Schult. and Mantisia wengeri Fischer. Highest Bud Forming Capacity (BFC) of 6.10±0.55 with an average of 19.93±3.19 roots was obtained for M. spathulata within 5-6 weeks in Murashige and Skoogs (MS) medium supplemented with a combination of 10.0µM of N6-benzyladenine (BA) and 2.5µM of α-naphtalene acetic acid (NAA). For M. wengeri, BFC of 7.82±0.73 and 20.86±1.65 roots was achieved in MS media supplemented with a combination of 5.0µM BA and 2.5µM of NAA RAPD markers were used to evaluate the genetic stability of in vitro raised hardened plantlets. Similarity coefficient among the regenerated plants ranged between 0.85-0.98 for M. spathulata and 0.83-0.98 for M. wengeri. Maximum of 88 and 90% genetic similarity were obtained between in vitro raised hardened plantlets and mother stock of M. spathulata and M. wengeri, respectively through RAPD analysis. The hardened plantlets after RAPD analysis on being transferred to soil of experimental garden showed no marked phenotypic variations in vegetative or floral characteristics.

 

 

Indian Journal of Experimental Biology

Vol. 47, February 2009, pp. 147-150

 

 

Notes

Effect of thyroxine on plasminogen activator and inhibitor activity in rat

P Bubber*, N Bubber  & D D Bansal

Department of Biochemistry, Panjab University,
Chandigarh, 160 014, India

Received 20 August 2008; revised 1 December 2008

Thyroid hormones influence mineral metabolism, distribution of water and electrolytes and are therefore of great importance in the maintenance of homeostasis under normal and diseased conditions such as renal failure. The present study was carried out to determine the effect of thyroxine on fibrinolytic parameters such as plasminogen activators (PA) in rat kidney, levels of PA and plasminogen activator inhibitor (PAI), glucose in plasma and serum lipid profile injected with thyroxine (75 µg eltroxine/100 g-1 body weight, ip for 7 days). Treatment increased PA activity significantly in rat kidneys. No changes were seen in PA, PAI and glucose in plasma of rats. There was significant decrease in total cholesterol and LDL-cholesterol levels in serum of treated group resulting in the decrease of HDL/LDL and total cholesterol/cholesterol ratios. However, triglycerides and VLDL showed significant higher activity in the serum of treated group as compared to controls.  The results suggest beneficial effects of thyroxine treatment by increasing PA activity in kidney and reducing the cholesterol content in blood. It may be helpful to prevent hypercoagulable state by maintaining the normal homeostatic balance and restoring renal function.