Indian Journal of Experimental Biology

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VOLUME 48

NUMBER 2

FEBRUARY 2010

CODEN: IJEB (A6) 48 (2) - (2010) 89-180

ISSN: 0019-5189 (Print); 0975-1009 (Online)

 

                                                                   CONTENTS

 

Review Article

 

Anticancer potential of animal venoms and toxins

93

      Antony Gomes, Pushpak Bhattacharjee, Roshnara Mishra, Ajoy K. Biswas, Subir Chandra Dasgupta, Biplab Giri, Anindita Debnath, Shubho Das Gupta, Tanaya Das & Aparna Gomes

 

 

 

Papers

 

Poly I:C induced microglial activation impairs motor activity in adult rats

104

      I K Patro, Amit, M Shrivastava, S Bhumika & N Patro

 

 

 

Ibal expressing microglia in the dorsal root ganglia become activated following peripheral nerve injury in rats

110

      Nisha Patro, Aarti Nagayach & I K Patro

 

 

 

Evaluation of Mycobacterium tuberculosis specific RD antigens for delayed type hypersensitivity responses in guinea pig

117

      Mamta Kalra, Gopal Krishen Khuller, Javaid Ahmad Sheikh & Indu Verma

 

 

 

Characteristics of phase 3-like activity and rebound excitation triggered by hexamethonium and atropine administration in the ovine small bowel

124

      K W Romański

 

 

 

Protective effect of proanthocyanidins on endotoxin induced experimental periodontitis in rats

133

      Jayamathi Govindaraj, Pamela Emmadi, Deepalakshmi, Vijayalakshmi Rajaram, Geetha Prakash & Rengarajulu Puvanakrishnan

 

 

 

Protective effect of ginger against alcohol-induced renal damage and antioxidant enzymes in male albino rats

143

      K R Shanmugam, C H Ramakrishna, K Mallikarjuna & K Sathyavelu Reddy

 

 

 

Ameliorative effect of a combination of vitamin E, vitamin C, a- liopic acid and stilbene resveratrol on lindane induced toxicity in mice olfactory lobe and cerebrum

150

      Mehajbeen Bano & Devendra Kumar Bhatt

 

 

 

2-Hydroxy 4-methoxy benzoic acid isolated from roots of Hemidesmus indicus ameliorates liver, kidney and pancreas injury due to streptozotocin-induced diabetes in rats

159

      Mahalingam Gayathri & Krishnan Kannabiran

 

 

Hypolipidaemic and antioxidant effects of fruits of Musa AAA (Chenkadali) in alloxan induced diabetic rats

165

      Smitha Kaimal, K S Sujatha & Sisilamma George

 

 

 

Production and partial characterization of collagenase of Streptomyces exfoliatus CFS 1068 using poultry feather

174

      Richa Jain & P C Jain

 

 

 

Book Review

 

Plant Tissue Culture and Molecular Markers—Their Role in Improving Crop Productivity

179

      A B Das, G R Rout & Prasaanna Mohanty

 

 

 

Announcement

 

National Conference on Impact of Climate Change on Biodiversity of Insects and Environment (CBI’ 10)

92

 

 

—————————

 

 

Announcement

 

National Conference on

Impact of Climate Change on Biodiversity of Insects and Environment (CBI’ 10)

15 and16 March 2010, Coimbatore

With a view to commemorate the International Year of Biodiversity, 2010, the Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore, is organizing the titled Conference at the Department. The themes of the Conference are:(i) Impact of Climate Change on Biodiversity, Bioinformatics and Insect Migration,(ii) Impact of Climate Change on Crop Pests and Food Production, (iii) Impact of Climate Change on Forestry Insects Diversity and Ecosystem, and (iv) Impact of Climate Change on Insect Vector and Disease Surveillance/Outbreak. The special theme of the conference will be Global Information System (GIS) and Remote Sensing (RS) and Bio-Geo-Information. For details, please contact Prof. K. Murugan, Organizing Secretary, CBI,’10, Department of Zoology, School of Life Sciences, Bharathiar University, Coimbatore 641 046, India.

Phone (Office): +91-422-2428 492, Ext: 492; (Home):+91-422-2425 015; Fax: +91-422-2422 387, 2425 706; Mobile: +91-9894832849.

E-mail: kmvvk@yahoo.com, kmvvk@rediffmail.com; kmvvkg@gmail.com Website: www.b-u.ac.in

 

Author Index

 

Amit

104

 

 

Bano Mehajbeen

150

Bhatt Devendra Kumar

150

Bhattacharjee Pushpak

93

Bhumika S

104

Biswas Ajoy K.

93

 

 

Das A B

180

Das Gupta Shubho

93

Das Tanaya

93

Dasgupta Subir Chandra

93

Debnath Anindita

93

Deepalakshmi

133

 

 

Emmadi Pamela

133

 

 

Gayathri Mahalingam

159

George Sisilamma

165

 

 

Giri Biplab

93

Gomes Antony

93

Gomes Aparna

93

Govindaraj Jayamathi

133

 

 

Jain P C

174

Jain Richa

174

 

 

Kaimal Smitha

165

Kalra Mamta

117

Kannabiran Krishnan

159

Khuller Gopal Krishen

117

 

 

Mallikarjuna K

143

Mishra Roshnara

93

Mohanty Prasaanna

180

 

 

Nagayach Aarti

110

 

 

Patro I K

104,110

Patro N

104

Patro Nisha

110

Prakash Geetha

133

Puvanakrishnan
Rengarajulu

 

133

 

 

Rajaram Vijayalakshmi

133

Ramakrishna CH

143

Reddy K Sathyavelu

143

Romański K W

124

Rout G R

180

 

 

Shanmugam K R

143

Sheikh Javaid Ahmad

117

Shrivastava M

104

Sujatha K S

165

 

 

Verma Indu

117

 

Keyword Index

 

Anticancer activity

93

Acute phase proteins

133

Alcohol

143

Anticholinergic drugs

124

Antioxidant

133,143,165

Azocoll

174

 

 

Cerebrum

150

Collagenase

174

Combination therapy

93

 

 

Diabetes

159

Diabetes mellitus

165

Diagnosis

117

Dorsal root ganglia

110

DTH response

117

Duodenum

124

 

 

Experimental periodontitis

133

 

 

Histopathology

143

2-Hydroxy 4-methoxy benzoic
acid

 

159

Hypolipidaemic

165

 

 

 

 

Ibal

110

Injury

110

 

 

Jejunum

124

 

 

Kidney

143

 

 

Lindane

150

Lipid peroxidation

159

a-lipoic acid

150

Lysosomal enzymes

133

 

 

Metronidazole

133

MHC II

110

Microglia

104,110

Migrating myoelectric
complex

 

124

Motor activity

104

Musa AAA (Chenkadali)

165

 

 

Olfactory lobe

150

Oxidative stress

143

Phase 3-like activity

124

Poly I:C

104

Poultry feathers

174

 

 

 

 

PPD

117

Proanthocyanidin

133

Protease

174

 

 

Rats

110,143,165

RD antigens

117

Reactive oxygen
species

 

133

Rebound excitation

124

 

 

Satellite glial cells

110

Sheep

124

Stilbene resveratrol

150

Streptomyces exfoliates
CFS 1068

 

174

Streptozoiticin

159

 

 

Toxins

93

Tuberculosis

117

 

 

Venoms

93

Viral infection

104

Vitamin C

150

Vitamin E

150

 

 

Correspondent author has been indicated by * sign

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 93-103

 

 

 

Review Article

 

 

 

Anticancer potential of animal venoms and toxins

Antony Gomes*, Pushpak Bhattacharjee, Roshnara Mishra, Ajoy K. Biswas, Subir Chandra Dasgupta &
Biplab Giri

Laboratory of Toxinology and Experimental Pharmacodynamics, Department of Physiology, University of Calcutta,
92 A P C Road, Kolkata 700 009, India

and

Anindita Debnath, Shubho Das Gupta, Tanaya Das & Aparna Gomes

Drug Development Division, Indian Institute of Chemical Biology, Kolkata 700 032, India

 

Anticancer drug development from natural resources are ventured throughout the world.  Animal venoms and toxins a potential bio resource and a therapeutic tool were known to man for centuries through folk and traditional knowledge. The biodiversity of venoms and toxins made it a unique source of leads and structural templates from which new therapeutic agents may be developed. Venoms of several animal species (snake, scorpion, toad, frog etc) and their active components (protein and non protein toxins, peptides, enzymes, etc) have shown therapeutic potential against cancer. In the present review, the anticancer potential of venoms and toxins from snakes, scorpions, toads and frogs has been discussed. Some of these molecules are in the clinical trials and may find their way towards anticancer drug development in the near future. The implications of combination therapy of natural products in cancer have been discussed.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 104-109

 

 

Papers

 

Poly I:C induced microglial activation impairs motor activity in adult rats

I K Patro1,2*, Amit1, M Shrivastava1, S Bhumika1 & N Patro1

1School of Studies in Neuroscience, Jiwaji University, Gwalior 474011, India

2School of Studies in Zoology, Jiwaji University, Gwalior 474011, India

Received 5 October 2009; revised 18 November 2009

Polyinosinic:polycytidic acid (poly I:C) is a synthetic double stranded RNA, which mimics with viral genome and mediates immune activation response similar to double stranded RNA virus infection into the brain. Microglial cells are the immune competent cells of the central nervous system having Toll like receptors-3 on their surface. Upon establishing that poly I:C infusion into the brain causes microgliosis by creating a viral infection model, the present study was designed to evaluate the effects of microglial activation following poly I:C infusion on motor activity. We infused 100µl of 1% solution of Poly I:C in TBE buffer directly into the lateral ventricle and TBE buffer as vehicle to controls. A significantly higher microglial cell count as compared to control on 2, 3 and 7 days post infusion was recorded. Motor activity and microglial cell count was assessed in both controls and poly I:C infused rats on 1,2,3,7,14,21 and 28 days post infusion. A significant decrease in motor activity and motor coordination occurred with respect to control. The results clearly demonstrate that microglial activation has a direct relevance with decreased motor activity. Findings could also have their importance in understanding the role of microglial cells on behavioral aspects in viral diseases.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 110-116

 

 

 

Iba1 expressing microglia in the dorsal root ganglia become activated following peripheral nerve injury in rats

Nisha Patro, Aarti Nagayach & I K Patro*

School of Studies in Neuroscience, Jiwaji University, Gwalior 474 011, India

Received 5 October 2009; revised 17 November 2009

The presence of microglia in dorsal root ganglia (DRG) has not been reported earlier. The dorsal root ganglia contain satellite glial cells (SGCs) and macrophages, which are considered to have infiltrated from the systemic blood. An attempt was made to investigate whether microglia as found in the central nervous system are also present in the dorsal root ganglia of untreated rats and following experimental peripheral nerve injury. Female adult Wistar rats were subjected to sciatic nerve transection injury on the right hand side. The DRGs of the right side were studied with the contralateral DRGs of the left side serving as controls. The tissues, harvested at different time points after injury, following intracardial perfusion fixation, and frozen sections were immunolabeled with anti-GFAP as a marker for SGCs and anti-Iba1 and OX-6 as markers for microglia and activated macrophagic microglia, respectively. These antibodies were also used in combination to ascertain if Iba1+ cells are the SGCs or otherwise and also if macrophagic OX-6+ cells are Iba1 positive microglia. The results indicate that Iba1 positive microglial cells are different from the SGCs in the DRGs. The Iba1 positive microglial cells respond to the sciatic nerve injury becoming activated and macrophagic and express MHCII molecules. Such activated microglia apparently may serve as neurosupportive cells, providing neuroprotection and scavenging cellular debris in response to the injury.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 117-123

 

 

Evaluation of Mycobacterium tuberculosis specific RD antigens for delayed
type hypersensitivity responses in guinea pig

Mamta Kalra, Gopal Krishen Khuller, Javaid Ahmad Sheikh & Indu Verma*

Department of Biochemistry, Post Graduate Institute of Medical Education and Research, Chandigarh 160012, India

Received 21 April 2009; revised 6 October 2009.

Tuberculin skin test (TST), an age old method is based on measuring delayed-type hypersensitivity (DTH) response to purified protein derivative (PPD). However, inspite of simplicity, ease and cost effectiveness, the usefulness of PPD test is limited due to its inability to distinguish among a protective immune response, latent infection and active tuberculosis disease. On the other hand, a skin test based on RD antigens would add advantages of a high specificity of antigens with the logistics of a skin test. However, except few reports, in vivo data of intradermal use of RD antigens for skin testing is limited. Therefore, in the present study, four M. tuberculosis (Mtb) specific antigens (ESAT6, CFP10, CFP21 and MPT64) were evaluated for their diagnostic utility based on DTH response. These antigens alone and their multiple combinations induced strong DTH response in Mtb infected guinea pigs and the response was negligible in BCG vaccinated and sham immunized animals.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 124-132

 

 

 

Characteristics of phase 3-like activity and rebound excitation triggered by hexamethonium and atropine administration in the ovine small bowel

K W Romański

Laboratory of Clinical Physiology, Department of Biostructure and Physiology, Faculty of Veterinary Medicine
Wroclaw University of Environmental and Life Sciences, Norwida 31, 50-375 Wrocław, Poland

Received 18 June 2009; revised 10 November 2009

Administration of hexamethonium (Hx) and atropine inhibits myoelectric and motor activity and then evokes a stimulatory effect called rebound excitation (RE) in the ovine small bowel. RE has not been precisely characterized so far and it is possible that it is composed of different types of motility. This study was thus devoted to characterizing these excitatory changes in the myoelectric and motor activity of the small bowel, particularly in the duodenum in conscious sheep. These alterations occurred in response to different intravenous doses of Hx and atropine administered alone or in combinations during various phases of the migrating myoelectric or motor complex (MMC) in the fasted and non-fasted sheep. Initially two basic types of excitatory response to the cholinergic blockade were found. In the course of chronic experiments different doses of Hx and atropine evoked phase 3-like activity (unorganized phase 3 of the MMC or its fragments) alternating with the less regular RE and the duration of these changes was related to the drug dose. In the non-fasted sheep these changes were less pronounced than in the fasted animals. When the drug was given during phase 1 of the MMC, RE did not occur or was greatly reduced. Administration of Hx and atropine in the course of phase 2a and phase 2b of the MMC produced roughly similar effects. Hx triggered stronger phase 3-like activity and RE than atropine. Combinations of Hx and atropine induced an additive effect, more evident in the fasted animals. These actions of Hx and atropine, thus, appear to involve at least partly the same intramural pathways. It is concluded that Hx and atropine evoke phase 3-like activity alternating with RE as the secondary stimulatory response in conscious sheep and both these types of the intestinal motility represent two distinct motility patterns.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 133-142

 

 

Protective effect of proanthocyanidins on endotoxin induced experimental periodontitis in rats

Jayamathi Govindaraj1, Pamela Emmadi2, Deepalakshmi2,

Vijayalakshmi Rajaram2, Geetha Prakash3 & Rengarajulu Puvanakrishnan4 *

Department of 1Biochemistry and2 Periodontics, Meenakshi Ammal Dental College, Alapakkam Main Road,

Maduravoyal, Chennai 600 095, India

3Department of Pathology, Meenakshi Medical College, Enathur, Kanchipuram 631552, India

4Department of Biotechnology, Central Leather Research Institute, CSIR, Chennai 600 020, India

Received 5 August 2009; revised 17 November 2009

The pathogenesis of periodontitis involves anaerobic oral bacteria as well as the host response to infection and several drugs have been developed which can curtail these deleterious effects. Proanthocyanidin, a novel flavanoid extracted from grape seeds, has been shown to provide a significant therapeutic effect on endotoxin (Escherichia coli) induced experimental periodontitis in rats. In this study, protective action of different doses of proanthocyanidins was investigated in blood by assaying the reactive oxygen species such as hydrogen peroxide, superoxide anion, myeloperoxidase and lipid peroxides, lysosomal enzyme activities such as cathepsin B, cathepsin D, β-glucuronidase and acid phosphatase, nonenzymatic antioxidants such as ascorbic acid, α-tocopherol, ceruloplasmin, reduced glutathione and antioxidant enzymes such as catalase, superoxide dismutase, glutathione peroxidase and glutathione-s-transferase. Experimental periodontitis rats showed a reduction in body weight and body weight gain could be noticed when they were administered proanthocyanidins. The levels of reactive oxygen species and lysosomal enzymes were found to increase whereas antioxidant levels were decreased significantly in experimental periodontitis. Proanthocyanidins at an effective dose of 30mg / kg body weight, sc, for 30 days effected a decrease in serum reactive oxygen species, lipid peroxides, lysosomal enzymes, acute phase proteins and an increase in antioxidant levels. Histopathological evidence of experimental periodontitis showed cellular infiltration of inflammatory cells while proanthocyanidin treated groups demonstrated only scattered inflammatory cells and blood vessels. Thus, the results showed that dietary supplementation of proanthocyanidin enhanced the host resistance as well as the inhibition of the biological and mechanical irritants involved in the onset of gingivitis and the progression of periodontal disease.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 143-149

 

 

Protective effect of ginger against alcohol-induced renal damage and antioxidant enzymes in male albino rats

K R Shanmugam, CH Ramakrishna, K Mallikarjunaa, & K Sathyavelu Reddy*

Division of Molecular Biology and Exercise Physiology, Department of Zoology,

Sri Venkateswara University, Tirupati 517 502, India

aLaboratory of Exercise Biochemistry, Taipei Physical Education College, Taipei 11153, Taiwan, ROC

Received 16 April 2009; revised 17 November 2009

Superoxide dismutase, ascorbic acid, glutathione and uric acid levels were decreased and xanthine oxidase, glutathione-s-transferase activities were increased in alcohol treated (2 g/kg body weight, once daily for 30 days) group. However, treatment with ethanolic extract of ginger (100 mg/kg, 200 mg/kg body weight, po, once daily for 30 days) these parameters came to normalcy showing the antioxidant effect of ginger. The antioxidant compounds of ginger may modulate the oxidative stress parameters. The biochemical findings were supplemented by histopathological examination of the kidney. Severe congestion and degenerative changes in tubules in alcohol treated rats were restored by ginger extract treatment. The results confirm the renal protective effect of ginger in alcohol treated rats.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 150-158

 

 

Ameliorative effect of a combination of vitamin E, vitamin C, α- lipoic acid and stilbene resveratrol on lindane induced toxicity in mice olfactory lobe and cerebrum

Mehajbeen Bano* & Devendra Kumar Bhatt

Cancer Biology and Toxicology laboratory, Department of Zoology

University College of Science, Mohan Lal Sukhadia University Udaipur 313 001, India

Received 16 June 2009; revised 9 November 2009

Acute dose of lindane (40 mg/kg body weight, ip) caused significant reduction in butyrylcholinesterase (BChE) activity both in olfactory lobe and cerebrum of mice along with reduction in catalase (CAT), total protein and elevation in superoxide dismutase (SOD) and cholesterol contents. Pretreatment by a combination of antioxidants, vitamin E, vitamin C, α- lipoic acid and stilbene resveratrol (125 mg/kg body weight, ip) significantly augment the altered level of BChE and protect the other parameters in both the brain regions. The results were adequately in agreement with the histochemical findings, suggesting the neuroprotective efficacy of combination of antioxidants studied on the lindane induced neurotoxicity.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 159-164

 

 

2-Hydroxy 4-methoxy benzoic acid isolated from roots of Hemidesmus indicus ameliorates liver, kidney and pancreas injury due to streptozotocin-induced diabetes in rats

Mahalingam Gayathri & Krishnan Kannabiran*

School of Biosciences and Technology, VIT University, Vellore 632 014,  India

Received 8 October 2008; revised 30 October 2009

Protective effect was evaluated in streptozoticin (STZ)-induced diabetes rats. 2-Hydroxy 4-methoxy benzoic acid (HMBA) was isolated from the roots of Hemidesmus indicus and administered (500 μg/kg body weight) orally for 7 weeks to STZ-induced diabetic and non-diabetic rats to study its effect on protein metabolism, serum electrolytes and on liver and kidney lipid peroxides. Oral administration of HMBA restored the altered biochemical parameters such as urea, uric acid, creatinine, plasma proteins and serum electrolytes to near-normal levels. HMBA treatment significantly decreased lipid peroxidation and malondialdehyde levels in diabetic liver and kidney. Effect of HMBA was equivalent to that of the standard drug, tolbutamide (100 mg/kg body wt). The histological changes were also in correlation with the biochemical findings. The present study showed that HMBA isolated from H. indicus roots had ameliorative effect on liver, kidney and pancreatic injury in STZ-induced diabetic rats.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 165-173

 

 

Hypolipidaemic and antioxidant effects of fruits of Musa AAA (Chenkadali) in alloxan induced diabetic rats

Smitha Kaimal, K S Sujatha1 & Sisilamma George*

Department of Veterinary Biochemistry, 1Department of Statistics, Faculty of Veterinary and Animal Sciences,
Mannuthy, Thrissur 680 651, India

Received 9 June 2009; revised 4 November 2009

Hypolipidaemic and antioxidant effects of ethanol extract of mature green fruits of Musa AAA (Chenkadali) was evaluated in alloxan induced diabetic rats. The effect of extract at two doses, 500 mg/kg body weight and 1000 mg/kg body weight was analysed and compared with a standard drug, glibenclamide. Rats administered with alloxan showed significantly increased levels of serum triacylglycerol, total cholesterol and alanine amino transferase (ALT) activity. Lipid peroxides increased significantly while reduced glutathione (GSH) decreased considerably in liver and pancreas. Oral administration of the ethanol extract of fruits of Musa AAA (Chenkadali) significantly decreased the levels of serum triacylglycerol, cholesterol and ALT activity. Significant decrease was also observed in the level of lipid peroxides while GSH content increased substantially in liver and pancreas. The effect was dose independent and rats treated with 500 mg/kg body weight showed comparable levels of serum triacylglycerol, cholesterol, ALT activity and liver lipid peroxides to that of normal control and glibenclamide treated groups. Although, there was no significant difference, treatment with
500 mg/kg body weight of the extract showed a higher content of GSH and lower level of lipid peroxides in pancreas compared with glibenclamide. Histopathological examination of pancreas and liver revealed regeneration of islet cells and hepatocytes respectively, which correlate with the biochemical findings. The present study shows that ethanol extract of mature green fruits of Musa AAA (Chenkadali) has antioxidant and hypolipidaemic properties and may be used for treating diabetes mellitus.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp. 174-178

 

 

Production and partial characterization of collagenase of Streptomyces exfoliatus CFS 1068 using poultry feather

Richa Jain & P C Jain*

Department of Applied Microbiology and Biotechnology, Dr. Hari Singh Gour University, Sagar 470 003, India

Received 2 April 2009; Revised 30 September 2009

Streptomyces exfoliatus CFS 1068, an isolate of cultivated field soil, produced maximum collagenase activity
(58.19
± 0.83 U ml-1min-1) in 5 days when soybean meal and starch were used as nitrogen and carbon sources, respectively at pH 7 and 30°C in shake cultures (150 rpm). Production of collagenase was higher (40.43± 0.63 U ml-1min-1) when poultry feathers were used as nitrogen source. Thus, the strain was found to be of biotechnological importance. The purified enzyme showed 30.34 fold increase in collagenase activity and was stable at 70°C for 1h. The enzyme was found to be of serine type.

 

 

Indian Journal of Experimental Biology

Vol. 48, February 2010, pp 179-180

 

 

Book Review

 

 

Plant Tissue Culture and Moleculr Markers—Their Role in Improving Crop Productivity