Indian Journal of Experimental Biology

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VOLUME 50

NUMBER 5

MAY 2012

CODEN: IJEB (A6) 50 (4) 309-378 (2012)

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

 

CONTENTS

 

Papers

 

Anti-HCMV and KSHV effect of a trapping ligand antagonist for Herpesvirus-encoded GPCR

Hong-Ai Liu, Han-Xiao Sun*, Xue-Mei Mo, Shi-Yu Li, Xiu-Ying Li, Guang Zhang & Lu Li

313

 

 

Nimodipine down-regulates CGRP expression in the rat trigeminal nucleus caudalis

Lakshmy Vijayan, Devanshu Bansal & Subrata Basu Ray

320

 

 

Prokaryotic expression of chicken infectious anemia apoptin protein and characterization of its

polyclonal antibodies

Shikha Saxena, Gandham Ravi Kumar, Prafull Singh, Uttara Chaturvedi, Lovleen Saxena,

Rajiv Kumar, Aditya Prasad Sahoo, Juwar Doley, Rajmani, Amit Kumar, Sudesh Kumar &

Ashok K Tiwari

325

 

 

Effect of supplementation of dermal fibroblasts conditioned medium on expansion of keratinocytes

through enhancing attachment

S R Chowdhury1, B S Aminuddin2 & B H I Ruszymah

332

 

 

Phenotypic and molecular characterization of indigenous rhizobia nodulating chickpea in India

Rhitu Rai, Prasanta K Dash, Trilochan Mohapatra & Aqbal Singh

340

 

 

Antihyperglycemic activity of Woodfordia fruticosa (Kurz) flowers extracts in glucose metabolism and lipid peroxidation in streptozotocin-induced diabetic rats

Neeraj Verma, G Amresh1, P K Sahu, Ch V Rao & Anil Pratap Singh

351

 

 

Antitumor potential of Castanopsis indica (Roxb. ex Lindl.) A. DC. leaf extract against Ehrlich's

ascites carcinoma cell

Narayan Dolai, Indrajit Karmakar, R B Suresh Kumar, Asis Bala, U K Mazumder &

Pallab Kanti Haldar

359

 

 

Effect of temperature on behavioural isolation : A study with Drosophila ananassae populations

Sujata Yadav & Anand Kumar Yadav

366

 

 

Nitric oxide alleviates oxidative damage induced by high temperature stress in wheat

A Bavita, B Shashi & S B Navtej

372

 

Editor痴 Note

 

The Indian Journal of Experimental Biology is covered by the following international abstracting and indexing services:

 

Science Citation Index ExpandedTM

PubMed (http://www.ncbi.nim.nih.gov/)

MEDLINE

BIOSIS

Chemical Abstracts Service

Excerpta Medica

Informascience

Refrativnyi Zhurnal

Zoological Records

 

 

 

 

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Indian Journal of Experimental Biology in Open Access Mode

 

The Indian Journal of Experimental Biology (IJEB) is now an open access journal in the repository, NISCAIR Online Periodicals Repository (NOPR) [http://nopr.niscair.res.in].

Full text of all articles published in IJEB from 2006 onwards can now be accessed at NOPR in the open access mode. Papers in the current issue shall be uploaded immediately. Papers published in earlier years shall be added soon.

NOPR is based on DSpace, a digital repository software, and allows document browsing, document searching and various search options like title, author name, keywords, year, issue, etc.

 

 

 

Author Index

Aminuddin B S

332

Amresh G

351

 

 

Bala Asis

359

Bansal Devanshu

320

Bavita A

372

 

 

Chaturvedi Uttara

325

Chowdhury S R

332

 

 

Dash Prasanta K

340

Dolai Narayan

359

Doley Juwar

325

 

 

Haldar Pallab Kanti

359

 

 

Karmakar Indrajit

359

Kumar Amit

325

Kumar Gandham Ravi

325

Kumar R.B.Suresh

359

Kumar Rajiv

325

Kumar Sudesh

325

 

 

Li Lu

313

Li Shi-Yu

313

Li Xiu-Ying

313

Liu Hong-Ai

313

 

 

Mazumder U. K.

359

Mo Xue-Mei

313

Mohapatra Trilochan

340

 

 

Navtej S B

372

 

 

Rai Rhitu

340

Rajmani

325

Rao Ch V

351

Ray Subrata Basu

320

Ruszymah B H I

332

Sahoo Aditya Prasad

325

Sahu P K

351

Saxena Lovleen

325

Saxena Shikha

325

Shashi B

372

Singh Anil Pratap

351

Singh Aqbal

340

Singh Prafull

325

Sun Han-Xiao

313

 

 

Tiwari Ashok K

325

 

 

Verma Neeraj

351

Vijayan Lakshmy

320

 

 

Yadav Anand Kumar

366

Yadav Sujata

366

 

 

Zhang Guang

313

 

Keyword Index

Antagonist

313

Antihyperglycemic

351

Antioxidant

351,359

Antioxidant enzymes

372

Antitumor

359

 

 

Behavioural isolation

366

 

 

Calcium channel blocker

320

cAMP response-element
 binding protein

320

Castanopsis indica

359

Chicken infectious anemia

325

Chickpea

340

Cytotoxicity

359

 

 

Dermal fibroblast conditioned medium

332

Diabetes mellitus

351

Drosophila ananassae

366

 

 

EAC cell

359

Efficiency of cell
 attachment

332

Expansion index

332

 

 

Genetic diversity

340

 

 

High temperature stress

372

Human cytomegalovirus

313

 

 

Image J

320

Immunohistochemistry

320

 

 

Kaposi痴 sarcoma-
 associated herpesvirus
 ORF74



313

Keratinocytes

332

 

 

Lipid peroxidation

372

 

 

Migraine

320

 

 

Peptide H22-LP

313

Phylogeny

340

Polyclonal sera

325

Prokaryotic expression

325

 

 

Rhizobia

340

 

 

Sodium nitroprusside

372

Streptozotocin

351

Symbiotic efficiency

340

 

 

Temperature effect

366

Tissue Engineering

332

Trapping receptor/ligand

313

Trigeminal ganglia

320

 

 

US28

313

 

 

VP3 gene

325

 

 

Wheat

372

Woodfordia fruticosa

351

 

 

Correspondent author has been indicated by * sign

 

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 313-319

 

 

Anti-HCMV and KSHV effect of a trapping ligand antagonist for
Herpesvirus-encoded
GPCR

Hong-Ai Liu, Han-Xiao Sun*, Xue-Mei Mo, Shi-Yu Li, Xiu-Ying Li, Guang Zhang & Lu Li

Institute of Genomic Medicine, College of Pharmacy, Jinan University, Guangzhou, 510632, China

Received 28 September 2011; revised 28 February 2012

We have found and synthesized a trapping ligand peptide H22-LP (the conservative sequence is NAHCALL) from a random phage library according to the broad-spectrum trapping receptor H22, which derived from the residue 14-35 near the N-terminal region of receptor US28 on HCMV. In this study, we will evaluate its potential as an efficient antagonist of US28 and the anti-virus activity, acting as a broad spectrum chemokine receptors antagonist. Stable expression of US28 and ORF74 in NIH/3T3 cells were successfully constructed in vitro. Flow cytomety was used to determine the concentration of Ca2+ induced by H22-LP, and the binding of H22-LP and US28 was confirmed by enzyme-linked immunosorbent assay (ELISA). Antivirus activity of H22-LP on HCMV and KSHV was evaluated by anti-virus experiments. Our data suggest that H22-LP is an effectual antagonist of receptor US28 of HCMV and ORF74 of KSHV in the transfection assay, and it has potential to inhibit infection of HCMV and KSHV. These results provide support for the development of anti-virus strategies based on targeted inhibiting the infection of herpesvirus.

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 320-324

 

 

Nimodipine down-regulates CGRP expression in the rat
trigeminal nucleus caudalis

 

Lakshmy Vijayan, Devanshu Bansal & Subrata Basu Ray*

Department of Anatomy, All India Institute of Medical Sciences (AIIMS), New Delhi 110 029, India

Received 15 July 2011; revised 6 February 2012

L-type calcium channel blockers like verapamil are used in the prophylaxis of migraine. However, their effect on the expression of CGRP in the trigeminal nucleus caudalis (TNC) is unknown. It is important because an earlier study had shown that olcegepant, a CGRP receptor antagonist, acts at the level of the trigeminal spinal nucleus rather than the trigeminal ganglia. Nimodipine was used in the present study as it crosses the blood-brain barrier. The objective of the study was to determine the pattern of expression of calcitonin gene-related peptide (CGRP) in the TNC after administration of nimodipine and/or morphine. Wistar rats were injected with saline, morphine, nimodipine or morphine + nimodipine for 14 days. Subsequently, the lowest part of the medulla oblongata containing the spinal nucleus was removed and processed for immunohistochemical localization of CGRP. The density of expression was quantified using Image J software. The results were statistically analyzed. CGRP expression was noted over the superficial part of the TNC, which decreased significantly after nimodipine administration. Conversely, morphine produced an up-regulation. The expression was unchanged with reference to saline in the morphine + nimodipine treated group. Decreased expression of CGRP in the trigeminal nucleus caudalis after nimodipine is being reported for the first time. Also, whether CGRP expression can be used as a marker for predicting the therapeutic efficacy of an anti-migraine drug is currently being investigated.

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 325-331

 

 

Prokaryotic expression of chicken infectious anemia apoptin protein and characterization of its polyclonal antibodies

Shikha Saxena, Gandham Ravi Kumar, Prafull Singh, Uttara Chaturvedi, Lovleen Saxena, Rajiv Kumar,
Aditya Prasad Sahoo , Juwar Doley, Rajmani, Amit Kumar, Sudesh Kumar & Ashok K Tiwari*

Molecular Biology Laboratory, Department of Veterinary Biotechnology,
Indian Veterinary Research Institute,
Izatnagar, 243 122, India

Received 2 June 201; revised 21 February 2012

In the present study recombinant VP3 (rVP3) was expressed in E.coli BL21 (DE3) (pLysS) and its polyclonal antibodies were characterized. SDS鳳AGE analysis revealed that the expression of recombinant protein was maximum when induced with 1.5 mM IPTG for 6 h at 37コC. The 6ラHis-tagged fusion protein was purified on Ni-NTA and confirmed by Western blot using CAV specific antiserum. Rabbits were immunized with purified rVP3 to raise anti-VP3 polyclonal antibodies. Polyclonal serum was tested for specificity and used for confirming expression of VP3 in HeLa cells transfected with pcDNA.cav.vp3 by indirect fluorescent antibody test (IFAT), flow cytometry and Western blot. Available purified rVP3 and polyclonal antibodies against VP3 may be useful to understand its functions which may lead to application of VP3 in cancer therapeutics.

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 332-339

 

 

Effect of supplementation of dermal fibroblasts conditioned medium on
expansion of keratinocytes through enhancing attachment

S R Chowdhury1, B S Aminuddin2 & B H I Ruszymah1,3*

1Tissue Engineering Centre, Universiti Kebangsaan Malaysia (National University of Malaysia) Medical Centre
Jalan Yaacob Latif, Bandar Tun Razak, Cheras, 56000 Kuala Lumpur, Malaysia

2Ear, Nose and Throat Consultant Clinic, Ampang Puteri Specialist Hospital, Jalan Mamanda 9,
Taman Dato Ahmad Razali, 68000 Selangor, Malaysia

3Department of Physiology, Universiti Kebangsaan Malaysia (National University of Malaysia) Medical Centre
Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia

Received 28 June 2011; revised 27 February 2012

In the present study in vitro expansion of human keratinocytes by supplementing dermal fibroblasts conditioned medium (DFCM) has been reported. Effect of two different DFCM acquired by culturing fibroblasts in keratinocyte-specific medium (defined keratinocytes serum free medium, DFCM-DKSFM) and fibroblast-specific serum free medium (F12: DMEM nutrient mix, DFCM-FD) have been compared. Growth kinetics of keratinocytes in terms of efficiency of cell attachment, expansion index, apparent specific growth rate and growth potential at the end of culture was evaluated in culture supplemented with DFCM-DKSFM and DFCM-FD in comparison with control i.e. DKSFM only. Results indicated that supplementation of DFCM caused significant increase in keratinocyte attachment. Efficiency of keratinocyte attachment in culture supplemented with DFCM-DKSFM was significantly higher compared to those cultured in DFCM-FD and DKSFM. In addition, the expansion index of keratinocytes in cultures supplemented with DFCM-DKSFM and DFCM-FD were 3.7 and 2.2 times higher than that of control condition even though the apparent growth rate and proliferative potential was found significantly lower. These results suggested that supplementation of DFCM enhanced expansion of keratinocyte by increasing efficiency of cell attachment, and DFCM-DKSFM provided suitable condition for in vitro expansion of keratinocytes compared to DFCM-FD and control condition.

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 340-350

 

 

Phenotypic and molecular characterization of indigenous rhizobia nodulating chickpea in India

Rhitu Rai*, Prasanta K Dash, Trilochan Mohapatra & Aqbal Singh

National Research Centre on Plant Biotechnology, Indian Agricultural Research Institute Campus, New Delhi 110 012, India

Received 20 July 2011; revised 6 February 2012

In a combined approach of phenotypic and genotypic characterization, 28 indigenous rhizobial isolates obtained
from different chickpea growing regions in peninsular and northern India were analyzed for diversity. The field isolates were compared to two reference strains TAL620 and UPM-Ca142 representing M. ciceri and M. mediterraneum respectively. Phenotypic markers such as resistance to antibiotics, tolerance to salinity, temperature, pH, phosphate solubilization ability, growth rate and also symbiotic efficiency showed considerable diversity among rhizobial isolates. Their phenotypic patterns showed adaptations of rhizobial isolates to abiotic stresses such as heat and salinity. Two salt tolerant strains (1.5% NaCl by T1 and T4) with relatively high symbiotic efficiency and two P-solubilising strains (66.7 and 71
mg/ml by T2 and T5) were identified as potential bioinoculants. Molecular profiling by 16S ribosomal DNA Restriction Fragment Length Polymorphism (RFLP) revealed three clusters at 67% similarity level. Further, the isolates were differentiated at intraspecific level by 16S rRNA gene phylogeny. Results assigned all the chickpea rhizobial field isolates to belong to three different species of Mesorhizobium genus. 46% of the isolates grouped with Mesorhizobium loti and the rest were identified as M. ciceri and M. mediterraneum, the two species which have been formerly described as specific chickpea symbionts. This is the first report on characterization of chickpea nodulating rhizobia covering soils of both northern and peninsular India. The collection of isolates, diverse in terms of species and symbiotic effectiveness holds a vast pool of genetic material which can be effectively used to yield superior inoculant strains.

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 351-358

 

 

Antihyperglycemic activity of Woodfordia fruticosa (Kurz) flowers
extracts in glucose metabolism and lipid peroxidation in
streptozotocin-induced diabetic rats

Neeraj Verma1*, G Amresh1, P K Sahu2, Ch V Rao3 & Anil Pratap Singh3

1Department of Pharmacology, Goel Institute of Pharmacy & Sciences,
Faizabad Road (Near Indira Canal), Lucknow 227 105, India

2Department of Pharmacology, School of Pharmaceutical Sciences, Siksha 前 Anusandhan University,
Kalinga Nagar, Ghatikia, Bhubaneswar 751 003, India

3Pharmacognosy and Ethnopharmacology Division, CSIR-National Botanical Research Institute,
Rana Pratap Marg, P.O. Box No. 436, Lucknow 226 001, India

Received 28 June 2011; revised 14 February 2012

The ethanolic extract of W. fruticosa flowers (250 and 500 mg/kg) significantly reduced fasting blood glucose level and increased insulin level after 21 days treatment in streptozotocin diabetic rats. The extract also increased catalase, superoxide dismutase, glutathione reductase, glutathione peroxidase activities significantly and reduced lipid peroxidation. Glycolytic enzymes showed a significant increase in their levels while a significant decrease was observed in the levels of the gluconeogenic enzymes in ethanolic extract treated diabetic rats. The extract has a favourable effect on the histopathological changes of the pancreatic β- cells in streptozotocin induced diabetic rats. The results suggest that W. fruticosa possess potential antihyperglycemic effect by regulating glucose homeostasis and antioxidant efficacy in streptozotocin-induced diabetic rats.

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 359-365

 

 

Antitumor potential of Castanopsis indica (Roxb. ex Lindl.) A. DC. leaf extract against Ehrlich's ascites carcinoma cell

Narayan Dolai1, Indrajit Karmakar2, R B Suresh Kumar3, Asis Bala2,3, U K Mazumder3 & Pallab Kanti Haldar2,3,*

1Department of Natural Products, National Institute of Pharmaceutical Education and Research, Kolkata 700 032, India.

2Department of Pharmacology, Himalayan Pharmacy Institute, Majhitar, East Sikkim 737 136, India.

3Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700 032, India.

Received 5 August 2011; revised 29 February 2012

Methanol extract of C. indica (MECI) leaves showed direct cytotoxicity on Ehrlich ascites carcinoma (EAC) cell in a dose dependant manner and there was significant decrease in the tumor volume, viable cell count, tumor weight and elevated the life span of EAC tumor bearing mice. Hematological profile and biochemical estimations were significantly restored to normal levels in MECI treated as compared to EAC control mice. MECI treatment significantly modulated the tissue antioxidant assay parameters as compared to the EAC control mice. The results revealed that MECI possesses significant dose dependent antitumor potential which may be due to its cytotoxicity and antioxidant properties.

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 366-371

 

 

Effect of temperature on behavioural isolation : A study with
Drosophila ananassae populations

Sujata Yadav* & Anand Kumar Yadav

Department of Zoology, Agra College, Agra 282 002, India

Received 1 August 2011; revised 17 February 2012

Experiments were conducted to study sexual isolation among two natural populations of Drosophila ananassae maintained at 18 C and 24 C for 12 generations in the laboratory to see the effect of this environmental variable on behavioural isolation. Multiple choice technique was used and matings were observed directly in Elens Wattiaux mating chamber. Results showed sexual isolation among strains that were maintained at different temperatures, indicating that temperature may have affected the mating behaviour of the flies which resulted in the induction of ethological isolation among the strains.

 

 

Indian Journal of Experimental Biology

Vol. 50, May 2012, pp. 372-378

 

 

Nitric oxide alleviates oxidative damage induced by high temperature
stress in wheat

A Bavita1*, B Shashi1 & S B Navtej2

1Department of Biochemistry, 2Department of Plant Breeding and Genetics
Punjab Agricultural University, Ludhiana, 141 004, Punjab

Received 27 September 2011; revised 18 February 2012

Effect of sodium nitroprusside (SNP), a donor of nitric oxide (NO) was examined in two wheat (Triticum aestivum L.) cultivars, C 306 (heat tolerant) and PBW 550 (comparatively heat susceptible) to study the extent of oxidative injury and activities of antioxidant enzyme in relation to high temperature (HT) stress. HT stress resulted in a marked decrease in membrane thermostability (MTS) and 2, 3, 5-triphenyl tetrazolium chloride (TTC) cell viability whereas content of lipid peroxide increased in both the cultivars. The tolerant cultivar C 306 registered less damage to cellular membranes compared to PBW 550 under HT stress. Activities of antioxidant enzymes viz, superoxide dismutase, catalase, ascorbate peroxidase, guaicol peroxidase and glutathione reductase increased with HT in both the cultivars. Following treatment with SNP, activities of all antioxidant enzymes further increased in correspondence with an increase in MTS and TTC. Apparently, lipid peroxide content was reduced by SNP more in shoots of heat tolerant cultivar C 306 indicating better protection over roots under HT stress. The up-regulation of the antioxidant system by NO possibly contributed to better tolerance against HT induced oxidative damage in wheat.