Indian Journal of Experimental Biology

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VOLUME 50

NUMBER 7

JULY 2012

CODEN: IJEB (A6) 50 (7) 379-442 (2012)

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

 

CONTENTS

 

Papers

 

 

 

Stress-responsive hypothalamic-nucleus accumbens regulation may vary depending on stressors

447

 

 

Seung June Noh, Dong-Won Kang, Sang Bae Yoo, Joo Young Lee, Jin Young Kim, Bom-Taeck Kim, Jong-Ho Lee & Jeong Won Jahng

 

 

 

Effect of ANXA2 gene single nucleotide polymorphism (SNP) on the development of osteonecrosis in Indian sickle cell patient: A PCR-RFLP approach

455

 

 

Sanjay Pandey, Ravi Ranjan, Sweta Pandey, Rahasya Mani Mishra, Tulika Seth & Renu Saxena

 

 

 

Production and characterization of monoclonal antibodies against a local isolate of classical swine fever virus

459

 

 

Dilip K Sarma, Namrata Kashyap, Pankaj Deka, Prabhat Medhi & Parimal Roychoudhury

 

 

 

Anticancer activity of sclerotiorin, isolated from an endophytic fungus Cephalotheca faveolata Yaguchi, Nishim. & Udagawa

464

 

 

Periyasamy Giridharan, Shilpa Amit Verekar, Amit Khanna, P D Mishra &Sunil Kumar Deshmukh

 

 

 

Asparagus racemosus Willd (Liliaceae) ameliorates early diabetic nephropathy in STZ induced diabetic rats

469

 

 

Rahul Somani, Abhay Kumar Singhai, Prashant Shivgunde & Dilpesh Jain

 

 

 

Anti-obese activity of Butea monosperma (Lam) bark extract in experimentally induced obese rats

476

 

 

Dixit P, T Prakash, Roopa Karki & D Kotresha

 

 

 

Carbohydrate and elicitor enhanced withanolide (withaferin A and withanolide A) accumulation in hairy root cultures of Withania somnifera (L.)

484

 

 

Madhavi Doma, Gauri Abhayankar, V D Reddy & P B Kavi Kishor

 

 

 

Degradation of bacterial DNA by a natural antimicrobial agent with the help of biomimetic membrane system

491

 

 

Suman Bhandary, Shaswati Chaki, Sayanti Mukherjee, Sukhen Das, Sanjit Mukherjee, Keya Chaudhuri & Sujata G Dastidar

 

 

 

Synergistic effect of calcium stearate and photo treatment on the rate of biodegradation of low density polyethylene spent saline vials

497

 

 

D Carol, S Karpagam , S J Kingsley & S Vincent

 

 

 

Biodiesel production from seed oil of Cleome viscosa L.

502

 

 

Rashmi Kumari, Vinod Kumar Jain & Sushil Kumar

 

 

 

Announcement

 

 

 

International Conference on Industrial Biotechnology (ICIB-2012), IX Convention of the Biotech Research Society, India, Indo-Italian Workshop on Food Biotechnology: Industrial Processing, Safety & Health

446

 

 

Information for Authors

511

 

覧覧覧覧覧

 

Announcement

 

International Conference on Industrial Biotechnology (ICIB-2012)

IX Convention of the Biotech Research Society, India

Indo-Italian Workshop on Food Biotechnology: Industrial Processing, Safety & Health

 

21-23 November 2012, Patiala, India

 

 

Jointly organized by Department of Biotechnology, Punjabi University, Patiala, The Biotech Research Society, India and the Embassy of Italy, New Delhi, the Conference along with IX Convention of the Biotech Research Society, India and Indo-Italian Workshop on Food Biotechnology will be held Punjabi University, Patiala. The Conference will have the following major thrust areas of Biotechnology and their relevance to the welfare of mankind: (i) Innovative fermentation, (ii) Medical biotechnology, (iii) Environmental biotechnology, (iv) Food biotechnology, (v) Agricultural biotechnology, (vi) Proteomics and Genomics, (vii) Novel enzymes and microorganisms, (viii) Process engineering and innovative downstream processing, and (ix) Biocatalysis and process design. For further details, please contact Prof. R. S. Singh, Convener, ICIB-2012, Department of Biotechnology, Punjabi University, Patiala 147 002, India. Telephone: +91 175 3046262, 3046263; Fax: +91 175 2283073; E-mail: icibpup2012@gmail.com. Website: http://www.icibpup.org/

 

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Author Index

Abhayankar Gauri

484

 

 

Carol D

497

Chaki Shaswati

491

Chaudhuri Keya

491

 

 

Das Sukhen

491

Dastidar Sujata G

491

Deka Pankaj

459

Deshmukh Sunil Kumar

464

Doma Madhavi

484

 

 

Giridharan Periyasamy

464

 

 

Jahng Jeong Won

447

Jain Dilpesh

469

Jain Vinod Kumar

502

 

 

Kang Dong-Won

447

Karki Roopa

476

Karpagam S

497

Kashyap Namrata

459

Kavi Kishor P B

484

Khanna Amit

464

Kim Bom-Taeck

447

Kim Jin Young

447

Kingsley S J

497

Kotresha D

476

 

 

Lee Jong-Ho

447

Lee Joo Young

447

 

 

Medhi Prabhat

459

Mishra P D

464

Mishra Rahasya Mani

455

Mukherjee Sanjit

491

Mukherjee Sayanti

491

 

 

Noh Seung June

447

 

 

P Dixit

476

Pandey Sanjay

455

Pandey Sweta

455

Prakash T

476

 

 

Ranjan Ravi

455

Rashmi Kumari

502

Reddy V D

484

Roychoudhury Parimal

459

 

 

Sarma Dilip K

459

Saxena Renu

455

Seth Tulika

455

Shivgunde Prashant

469

Singhai Abhay Kumar

469

Somani Rahul

469

Suman Bhandary

491

Sushil Kumar

502

 

 

Verekar Shilpa Amit

464

Vincent S

497

 

 

Yoo Sang Bae

447

 

Keyword Index

Agarose gel electrophoresis


491

Antibacterial activity

491

Anticancer activity

464

ANXA2

455

Arthrobacter globiformis

497

Arthrobacter oxydans

497

Asparagus racemosus

469

Atherogenic diet

476

 

 

BAX

464

BCL-2

464

Biodegradation

497

Butea monosperma

476

 

 

Cafeteria diet

476

Calcium stearate

497

Caspase-3

464

Cephalotheca faveolata

464

Classical swine fever virus


459

Cleome viscosa seed oil

502

 

 

Diabetic nephropathy

469

 

 

Elicitors

484

Endophytic fungus

464

Eugenia jambolana

464

 

 

Fluorescence anisotropy

491

 

 

Hairy roots

484

Hypothalamus

447

 

 

Linoleic acid rich oil

502

Liposomal membrane

491

 

 

Microalbuminuria

469

Monoclonal antibodies

459

Monosodium glutamate

476

 

 

Non edible biodiesel oil

502

Nucleus accumbens

447

 

 

Obesity

476

 

 

PCR-RFLP

455

Plastic

497

Pro-oxidant solution

497

 

 

Restriction digestion

455

 

 

SNPs

455

Soybean/sunflower like oil


502

Streptozotocin

469

Stress

447

 

 

UV irradiation

497

 

 

van稚 Hoff enthalpy change


491

 

 

Western blotting

459

Withania somnifera

484

Withasteroids

484

 

 

Correspondent author has been indicated by * sign

 

 

Papers

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 447-454

 

 

 

Stress-responsive hypothalamic-nucleus accumbens regulation may vary depending on stressors

Seung June Noh1#, Dong-Won Kang2#, Sang Bae Yoo1, Joo Young Lee1, Jin Young Kim1, Bom-Taeck Kim3,
Jong-Ho Lee1** & Jeong Won Jahng1*

1Dental Research Institute, Department of Oral and Maxillofacial Surgery, Seoul National University School of Dentistry,
Seoul, 110-768, Korea;

2Department of Pharmacology, Kwandong University College of Medicine, Gangneung, 210-701, Korea;

3Department of Family Practice, Ajou University College of Medicine, Suwon, 443-721, Korea

Received 3 February 2012

This study was conducted to determine if the stress-responsive hypothalamic-nucleus accumbens (NAc) regulation is a stressor specific event. Male SD rats were subjected to restraint or cold stress for 2 h, and then mRNA expression of corticotropin-releasing hormone (CRH) in the hypothalamic paraventricular nucleus (PVN) was examined by in situ hybridization and the plasma corticosterone levels by radioimmunoassay. Neuronal activations in the PVN and the NAc were examined by c-Fos immunohistochemistry and the brain GABA contents by HPLC. Both restraint and cold stresses increased c-Fos expression in the PVN and the plasma corticosterone; however, CRH expression in PVN was increased only by restraint, but not by cold, stress. Restraint stress significantly increased the NAc neuronal activation, but cold stress failed to do so. Restraint stress increased the NAc-GABA contents and cold stress did the hypothalamic GABA. Results suggest that the HPA axis regulation responding to restraint stress, but not cold stress, may involve the NAc neuronal activation in relation with GABAergic neurotransmission. Additionally, CRH expression in the PVN may not play a major role in the elevation of plasma corticosterone responding to cold stress.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 455-458

 

 

Effect of ANXA2 gene single nucleotide polymorphism (SNP)
on the development of osteonecrosis in Indian sickle cell patient:
A PCR-RFLP approach

 

Sanjay Pandey1, Ravi Ranjan1, Sweta Pandey1, Rahasya Mani Mishra2, Tulika Seth1 & Renu Saxena1,*

1Department of Hematology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110 029, India

2 Department of Environmental Biology, Awadhesh Pratap Singh University, Rewa 486 003, India

Received 16 December 2011; revised 23 April 2012

Osteonecrosis is a serious complication in sickle cell patients. The common sites of the necrosis are femoral head, head of the humerus and acetabulam. Annexin A2 (ANXA2) protein mainly functions in bone formation and bone resorption. Alteration of ANXA2 gene may affect the manifestations of osteonecrosis in the patients. PCR-RFLP is a common applicable technique for the detection of known mutation/polymorphisms. Here we are presenting application of the PCR-RFLP technique for determination of the ANXA2 gene single nucleotide polymorphism frequency and their clinical association among Indian sickle cell patients. Five known SNPs of ANXA2 gene (rs7170178, rs73435133, rs73418020, rs72746635 and rs73418025) were determined using the HpyCH4V, DdeI, HpyCH4III and Sau 961 restriction enzyme respectively. Restriction enzyme DdeI was common for rs73435133 and rs72746635 SNP. Only the rs7170178 SNP was detected among patient and control and the other four SNPs were absent in the studied groups. The frequency of ANXA2 gene rs7170178 SNP (A/G, G/G) was comparatively higher in sickle cell patients than controls and it was clinically associated with sickle cell osteonecrosis. The P value of heterozygotes (A/G) and homozygotes (G/G) genotypes were <0.001 and 0.001 respectively, which were highly significant. This study established the application of PCR-RFLP in detection of ANXA2 SNPs in sickle cell patients.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 459-463

 

 

Production and characterization of monoclonal antibodies against a local
isolate of classical swine fever virus

Dilip K Sarma*, Namrata Kashyap, Pankaj Deka, Prabhat Medhi & Parimal Roychoudhury

National Fellow Project Laboratory, Department of Microbiology, College of Veterinary Science,
Assam Agricultural University, Khanapara, Guwahati 781 022, India

Received 16 November 2011; revised 23 April 2012

Monoclonal antibodies (mAbs) against a classical swine fever virus (CSFV; subgenogroup 1:1) isolate from Assam, India were produced and characterized. Four fusions of myeloma cells (SP2/0Ag) were made with spleenocytes of 8-10 weeks old BALB/C mice immunized with the viral antigen. Several hybridoma clones secreting antibodies to the virus were obtained after four fusions, but five hybridoma clones secreting antibody specific to the virus could be stabilized. All the mAbs belong to the IgG2a isotype. Except one, none of the four mAbs showed cross reaction with bovine viral diarrhoea virus and border disease virus (BDV). One mAb showed cross reaction with BDV. All the four mAbs specific to CSFV showed reactivity with the parental virus in immunoperoxidase test (IPT) and with a single protein band (molecular weight 55 kD approximately) of the virus in western blotting. In neutralization peroxidase linked assay (NPLA) all the mAbs reacted with 13 CSFV local isolates as well as with the cell culture adapted lapinized vaccine virus strain belonging to the subgenogroup 1:1. This is the first report on production and characterization of mAbs against CSFV in India.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 464-468

 

 

 

Anticancer activity of sclerotiorin, isolated from an endophytic fungus Cephalotheca faveolata Yaguchi, Nishim. & Udagawa

Periyasamy Giridharan, Shilpa Amit Verekar, Amit Khanna, P D Mishra & Sunil Kumar Deshmukh*

Piramal Healthcare Ltd, 1, Nirlon Complex, Off Western Express Highway,
Near NSE Complex, Goregaon (East), Mumbai 400 063, India

Received 20 October 2011; revised 12 April 2012

Biodiversity provides critical support for drug discovery. A significant proportion of drugs are derived, directly or indirectly, from biological sources. Through high throughput screening (HTS) and bioassay-guided isolation, bioactive compound sclerotiorin has been isolated from an endophytic fungus Cephalotheca faveolata. Sclerotiorin was found to be potent anti-proliferative against different cancer cells. In this study sclerotiorin has been found to induce apoptosis in colon cancer (HCT-116) cells through the activation of BAX, and down-regulation of BCL-2, those further activated cleaved caspase-3 causing apoptosis of cancer cells.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 469-475

 

 

Asparagus racemosus Willd (Liliaceae) ameliorates early diabetic
nephropathy in STZ induced diabetic rats

 

Rahul Somania*, Abhay Kumar Singhaib, Prashant Shivgundec & Dilpesh Jainc

aDepartment of Pharmacology, Smt. Kashibai Navale College of Pharmacy, Pune 411 048, India

bDepartment of Pharmaceutical Sciences, Dr. H S Gour University, Sagar 470 003, India

cDepartment of Pharmacology, Sinhgad College of Pharmacy, Pune 411 041, India

Received 8 November 2011; revised 24 May 2012

Diabetic nephropathy is a major 荘microvascular鋳 complication of diabetes, differs from other causes of chronic kidney diseases in its predictability, with well-defined functional progression from hyperfiltration to micro- to macroalbuminuria to renal failure. The present study was undertaken to investigate the effect of Asparagus racemosus Willd (Liliaceae) on streptozotocin -induced early diabetic nephropathy. Single i.p injection of streptozotocin (55 mg/kg) was administered to induce early diabetic nephropathy in Wistar rats and thereafter treated orally with ethanolic extract of Asparagus racemosus (EEAR) at a dose level of 100 and 250 mg/kg daily for 4 weeks. The efficacy of extract was compared with diabetic control rats. A. racemosus treatment significantly decreased plasma glucose, creatinine, urea nitrogen, total cholesterol and triglyceride levels. Renal hypertrophy, polyuria, hyperfiltration, microalbuminuria and abnormal changes in the renal tissue as well as oxidative stress were effectively attenuated by EEAR treatment. Basement membrane thickening and mesangial proliferation formation without nodules were seen in diabetic rats, whereas these structural changes were reduced in EEAR treated groups. Results of this study suggested that A. racemosus has beneficial effect in the treatment of diabetic nephropathy.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 476-483

 

 

Anti-obese activity of Butea monosperma (Lam) bark extract in experimentally induced obese rats

Dixit P1, T Prakash2,*, Roopa Karki3 & D Kotresha4

1Department of Pharmacology, Acharya & B.M. Reddy College of Pharmacy, Bangalore 560 090, India

2Department of Pharmacology and, 3Pharmaceutics, Malla Reddy College of Pharmacy, Hyderabad 500 014, India

4Department of Biochemistry, Indian Institute of Science, Bangalore 560 012, India

Received 7 March 2011; revised 3 May 2012

To study the efficacy of ethanolic extract of B. monosperma bark in cafeteria and atherogenic diet fed rats and monosodium glutamate (MSG) obese rats, different doses (200, 400 and 800 mg/kg) of ethanolic extract of B. monosperma bark showed dose dependent decrease in body weight, daily food intake, glucose, lipids, internal organs weight and fat pad weight in cafeteria and atherogenic diet fed rats and monosodium glutamate obese rats. The results suggested that B. monosperma has significant anti-obese activity.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 484-490

 

 

Carbohydrate and elicitor enhanced withanolide (withaferin A and withanolide A) accumulation in hairy root cultures of Withania somnifera (L.)

Madhavi Doma1, Gauri Abhayankar2, V D Reddy2 & P B Kavi Kishor1*

1Department of Genetics, Osmania University, Hyderabad 500 007, India

2Center for Plant Molecular Biology, Osmania University, Hyderabad 500 007, India

Received 28 September 2011; revised 27 March 2012

Leaves of Withania somnifera contained more withaferin A and withanolide A than roots indicating that these compounds mainly accumulate in leaves. With an increase in age of the plant, withaferin A was enhanced with a corresponding decrease in withanolide A. Hairy root cultures were induced from leaf explants using Agrobacterium rhizogenes and the transgenic nature of hairy roots was confirmed by partial isolation and sequencing of rolB gene, which could not be amplified in untransformed plant parts. In hairy roots, withaferin A accumulated at 2, 3 and 4% but not at 6% sucrose, the highest amount being 1733 mg/g dry weight at 4% level. High and equal amounts of withaferin A and withanolide A accumulated (890 and 886 mg/g dry tissue respectively) only at 3% sucrose. Increasing concentrations of glucose enhanced withaferin A and it peaked at 5% level (3866 mg/g dry tissue). This amount is 2842 and 34% higher compared to untransformed roots and leaves (collected from 210-day-old plants) respectively. Withanolide A was detected at 5% glucose but not at other concentrations. While chitosan and nitric oxide increased withaferin A, jasmonic acid decreased it. Acetyl salicylic acid stimulated accumulation of both withaferin A and withanolide A at higher concentrations. Triadimefon, a fungicide, enhanced withaferin A by 1626 and 3061% (not detected earlier) compared to hairy and intact roots respectively.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 491-496

 

 

Degradation of bacterial DNA by a natural antimicrobial agent with the
help of biomimetic membrane system

Suman Bhandary1, #, Shaswati Chaki 2, #, Sayanti Mukherjee2, Sukhen Das1, Sanjit Mukherjee3,
Keya Chaudhuri3 & Sujata G Dastidar2,*

1Department of Physics, Jadavpur University, Raja S. C. Mullick Road, Kolkata 700 032, India

2Department of Microbiology, Herbicure Healthcare Bio-Herbal Research Foundation,
7 & 8 Metro Garden City, D.H. Road, Pailan, Kolkata 700 104, India

3Division of Molecular and Human Genetics, CSIR- Indian Institute of Chemical Biology,
Jadavpur, Kolkata 700 032, India

Received 15 December 2011; revised 10 April 2012

The antimicrobial efficacy of methylglyoxal (MG) against several gram-negative bacteria including Escherichia coli has been reported. To determine the mechanism of action of MG, molecular interactions between lipid and MG within the liposomal membrane were also investigated. Multilamellar and unilamellar vesicles were prepared from 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). The effect of MG on DPPC liposomal membrane was studied by fluorescence spectroscopy and differential scanning calorimetry. The results indicate that MG interacts mainly with the DPPC head group that produces a significant increase in the fluidity of liposomal vesicles, which could be the cause of a fusion/aggregation effect in microbial cells. The agarose gel electrophoresis study with the genomic DNA extracted from E. coli ATCC 25922 revealed that addition of MG could completely degrade this DNA within 1 h, pointing out to their distinctly high degree of sensitivity towards MG. Further, the drug was able to cross the cell membranes, penetrating into the interior of the cell and interacting with DNA for demonstrating antibacterial activity of MG.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 497-501

 

 

Synergistic effect of calcium stearate and photo treatment on the rate of biodegradation of low density polyethylene spent saline vials

D Carol1,2*, S Karpagam2, S J Kingsley1 & S Vincent2

1Department of Plant Biology and Biotechnology,
2 P.G. & Research Department of Advanced Zoology and Biotechnology, Loyola College
Chennai 600 034, India

Received 17 October 2011; revised 23 April 2012

The biodegradation of spent saline bottles, a low density polyethylene product (LDPE) by two selected Arthrobacter sp. under in vitro conditions is reported. Chemical and UV pretreatment play a vital role in enhancing the rate of biodegradation. Treated LDPE film exhibits a higher weight loss and density when compared to untreated films. Arthrobacter oxydans and Arthrobacter globiformis grew better in medium containing pretreated film than in medium containing untreated film. The decrease in density and weight loss of LDPE was also more for pretreated film when compared to untreated film indicating the affect of abiotic treatment on mechanical properties of LDPE. The decrease in the absorbance corresponding to carbonyl groups and double bonds that were generated during pretreatment suggest that some of the double bonds were cut by Arthrobacter species. Since Arthrobacter sp. are capable of degrading urea, splitting of urea group were also seen in FTIR spectrum indicating the evidence of biodegradation after microbial incubation. The results indicated that biodegradation rate could be enhanced by exposing LDPE to calcium stearate (a pro-oxidant) which acts as an initiator for the oxidation of the polymers leading to a decrease of molecular weight and formation of hydrophilic group. Therefore, the initial step for biodegradation of many inert polymers depends on a photo-oxidation of those polymers. The application in sufficient details with improved procedures utilizing recombinant microorganism with polymer degradation capacity can lead to a better plastic waste management in biomedical field. The present plastic disposal trend of waste accumulation can be minimized with this promising eco-friendly technique.

 

 

Indian Journal of Experimental Biology

Vol. 50, July 2012, pp. 502-510

 

 

Biodiesel production from seed oil of Cleome viscosa L.

Rashmi Kumari1, 2, Vinod Kumar Jain1 & Sushil Kumar*2

1 School of Environmental Sciences, Jawaharlal Nehru University (JNU) and 2Genetic Genomics Laboratory,
National Institute of Plant Genome Research (NIPGR), New Delhi 110 067, India

Received 21 November 2011; revised 9 April 2012

Edible oil seed crops, such as rapeseed, sunflower, soyabean and safflower and non-edible seed oil plantation crops Jatropha and Pongamia have proved to be internationally viable commercial sources of vegetable oils for biodiesel production. Considering the paucity of edible oils and unsustainability of arable land under perennial plantation of Jatropha and Pongamia in countries such as India, the prospects of seed oil producing Cleome viscosa, an annual wild short duration plant species of the Indogangetic plains, were evaluated for it to serve as a resource for biodiesel. The seeds of C. viscosa resourced from its natural populations growing in Rajasthan, Haryana and Delhi areas of Aravali range were solvent extracted to obtain the seed oil. The oil was observed to be similar in fatty acid composition to the non-edible oils of rubber, Jatropha and Pongamia plantation crops and soybean, sunflower, safflower, linseed and rapeseed edible oil plants in richness of unsaturated fatty acids. The Cleome oil shared the properties of viscosity, density, saponification and calorific values with the Jatropha and Pongamia oils, except that it was comparatively acidic. The C. viscosa biodiesel had the properties of standard biodiesel specified by ASTM and Indian Standard Bureau, except that it had low oxidation stability. It proved to be similar to Jatropha biodiesel except in cloud point, pour point, cold filter plugging point and oxidation stability. In view of the annual habit of species and biodiesel quality, it can be concluded that C. viscosa has prospects to be developed into a short-duration biodiesel crop.