Indian Journal of Experimental Biology

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VOLUME 52

NUMBER 4

APRIL 2014

CODEN: IJEB (A6) 52 (4) 291-382 (2014)

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

CONTENTS

 

 

Papers

 

 

 

Analysis of rapamycin induced autophagy in Dictyostelium discoideum

295

 

 

Pynskhem Bok Swer, Rakhee Lohia & Shweta Saran

 

 

 

Neuronal differentiation of PC12 and embryonic stem cells in two- and three-dimensional
in vitro culture

305

 

 

Soheil Sadri, Mozafar Khazaei, Ali Ghanbari, Mohammad Rasool Khazaei & Palak Shah

 

 

 

Epigallocatechin gallate induces the steady state mRNA levels of pS2 and PR genes in
MCF-7 breast cancer cells

312

 

 

Mohan C Manjegowda, Gauri Deb & Anil M Limaye

 

 

 

Chronic spinal infusion of loperamide alleviates postsurgical pain in rats

317

 

 

Rakesh Kumar, K H Reeta & Subrata Basu Ray

 

 

 

Protective effect of Azadirachta indica A. Juss against doxorubicin-induced cardiac toxicity
in tumour bearing mice

323

 

 

Ashwani Koul, Renu Goyal & Sanjay Bharati

 

 

 

Impairment of renal structure and function following heterogeneous chemical mixture
exposure in rats

332

 

 

Kiran Morya & Kauresh D Vachhrajani

 

 

 

Phenotypic plasticity and ecotypic variations in growth and flowering time of
Arabidopsis thaliana (L.) under different light and temperature conditions

344

 

 

Shubhangi Moharekar (Lokhande), Sanjay Moharekar, Tsuyoshi Kobayashi, Hiroaki Ishii,
Akihiro Sumida & Toshihiko Hara

 

 

Regulation of nitrogen metabolism in salt tolerant and salt sensitive Frankia strains

352

 

 

Amrita Srivastava & Arun Kumar Mishra

 

 

Biosynthesis of silver nanoparticles using fresh extracts of Tridax procumbens Linn

359

 

 

Himakshi Bhati訪ushwaha & C P Malik

 

 

 

Efficacy of Spodoptera litura multiple nucleopolyhedrovirus after serial passage through the homologous insect larval host

369

 

 

Mudasir Gani, R K Gupta & K Bali

 

 

 

Pharmacophore based approach to design inhibitors in Crustaceans: An insight into
the molt inhibition response to the receptor guanylyl cyclase

375

 

 

Sajal Shrivastava & S Adline Princy

 

 

覧覧覧覧覧覧覧

NISCAIR Policy on Plagiarism

 

The system of formal communication in science through publication in primary journals is based on originality and quality of information being the only criteria for publication. However, there have been tendencies to misuse the system and vitiate the process of science communication for personal benefits. One of the ills afflicting science communication is plagiarism. Attempts at plagiarism may range from verbatim, copying of extensive material of other authors, misappropriating results/data of others with minor changes in language/presentation without giving credit to original source, to publish essentially the same information more than once.

As the premier publisher in India of primary scientific journals in various disciplines of science and technology, NISCAIR strongly reiterates its policy of discouraging plagiarism of all kinds. All efforts are made to detect and frustrate attempts at plagiarism through editorial screening and rigorous peer review in respect of communications received for publication in NISCAIR publications. Cooperation of the scientific community is sought in our efforts to frustrate all attempts at plagiarism.

It is mandatory on the part of the corresponding author to furnish the following certificate at the time of submission of the manuscript for publication:

 

[This is to certify that the reported work in the article entitle, (give full title with all the authors name) submitted for publication in the journal, the. is an original one and has not been submitted for publication elsewhere. I/we further certify that proper citation to the previously reported work have been given and no data/table/figures have been quoted verbatim from other publications without giving due acknowledgement and without the permission of the author(s). The consent of all the authors of this article has been obtained for submitting the article to the journal, 套.

Signatures and names of all the authors]

 

In case any attempt to plagiarize is brought to our attention accompanied with convincing evidence, following steps would be taken:

(a)            After consulting the respective Editorial Board Members, authors guilty of plagiarism will be debarred from publishing their papers in NISCAIR journals

(b)           Heads of the departments/institutes of the offending authors will be intimated of such incidences of plagiarism.

(c)            Such incidents of plagiarism will be publicized through the concerned NISCAIR journals in consultation with the respective Editorial Board Members.

 

覧覧覧覧覧覧覧

 

 

Author Index

Bali K

369

Bharati Sanjay

323

Bhati訪ushwaha Himakshi

359

 

 

Deb Gauri

312

 

 

Gani Mudasir

369

Ghanbari Ali

305

Goyal Renu

323

Gupta R K

369

 

 

Hara Toshihiko

344

 

 

Ishii Hiroaki

344

 

 

Khazaei Mohammad Rasool

305

Khazaei Mozafar

305

Kobayashi Tsuyoshi

344

Koul Ashwani

323

 

 

Limaye Anil M

312

Lohia Rakhee

295

 

 

Malik C P

359

Manjegowda Mohan C

312

Mishra Arun Kumar

352

Moharekar (Lokhande) Shubhangi

344

Moharekar Sanjay

344

Morya Kiran

332

 

 

Princy S Adline

375

 

 

Rakesh Kumar

317

Ray Subrata Basu

317

Reeta K H

317

 

 

Sadri Soheil

305

Saran Shweta

295

Shah Palak

305

Shrivastava Sajal

375

Srivastava Amrita

352

Sumida Akihiro

344

Swer Pynskhem Bok

295

 

 

Vachhrajani Kauresh D

332

 

 

 

Keyword Index

Antimicrobial

359

Apoptosis

305

Arabidopsis thaliana

344

Autophagy

295

Azadirachta indica

323

 

 

Baculoviruses

369

Bioinsecticides

369

Biosynthetic approach

359

Breast cancer

312

 

 

Calcium

295

Carcinogenesis

312

Cardiac toxicity

323

 

 

3D culture

305

Dictyostelium

295

Doxorubicin

323

 

 

Ecdysteroid

375

Embryonic stem cells

305

Entomopathogens

369

Epigallocatechin gallate

312

Estrogen responsive markers

312

 

 

Fibrin gel

305

Flowering

344

Frankia

352

FTIR

359

 

 

Growth enhancers

375

 

 

Hargreaves test

317

Heterogeneous chemical mixture

332

HipHop

375

 

 

Intrathecal catheterization

317

 

 

Kidney structure and function

332

 

 

Latitude

344

Lipid peroxidation

332

Loperamide

317

Low doses

332

 

 

MCF-7

312

Molt inhibiting hormone

375

Morphine

317

 

 

Nanoparticles

359

Nitrogen metabolism

352

Noctuidae

369

Osmotic minipump

317

Oxidative stress

323,332

 

 

PC12 cells

305

Pharmacophore

375

Phenotypic plasticity

344

Plantar incision

317

Progesterone receptor

312

pS2

312

 

 

Rapamycin

295

ROS

295

 

 

Salinity

352

Salt sensitive

352

Salt tolerant

352

 

 

Temperature

344

Trefoil Factor 1 (pS2)

312

Tridax procumbens

359

Tumour

323

 

 

von Frey test

317

 

 

Correspondent author is marked by *

 

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 295-304

 

 

Analysis of rapamycin induced autophagy in Dictyostelium discoideum

Pynskhem Bok Swer, Rakhee Lohia1 & Shweta Saran*

School of Life Sciences, Jawaharlal Nehru University, New Delhi 110 067, India

1Department of Genetics, University of Delhi, South Campus, New Delhi 110 021, India

Received 9 July 2013; revised 11 December 2013

Natural autophagy and autophagic cell death is being studied in the model system, D. discoideum, which has well known genetic and experimental advantages over the other known systems. There is no apoptotic machinery present in this organism which could interfere with the non-apoptotic cell death. The target of rapamycin (TOR) pathway is a major nutrient-sensing pathway which when inhibited by the drug rapamycin induces autophagy. Rapamycin was originally discovered as an anti-fungal agent but its use was abandoned when it was discovered to have potent immunosuppressive and anti-proliferative properties. It is a known drug used today for various cancer treatments and also for increasing longevity in many model organisms. It has a wide usage but its effects on other pathways or molecules are not known. This model system was used to study the action of rapamycin on autophagy induction. Using the GFP-Atg8, an autophagosome marker, it was shown that rapamycin treatment can induce autophagy by an accumulation of reactive oxygen species and intracellular free calcium. Rapamycin suppresses proliferation by induction of cell cycle arrest in the G1 phase. Taken together, the results suggest that the core machinery for autophagy is conserved in D. discoideum and it can serve as a good model system to delineate the action of rapamycin induced autophagy.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 305-311

 

 

Neuronal differentiation of PC12 and embryonic stem cells in two- and
three-dimensional in vitro culture

Soheil Sadri1,2, Mozafar Khazaei1,*, Ali Ghanbari1, Mohammad Rasool Khazaei1 & Palak Shah3

1Fertility and Infertility Research Center, Stem Cell Division, Kermanshah University of Medical Science,
University Avenue, Shahid Shiroodi Blvd, 6714869914, Kermanshah, Iran

2Division of Genomic Medicine, George Washington University, Washington DC, USA

3Division of Cardiology, George Washington University, Washington DC, USA

Received 14 November 2012; revised 11 June 2013

The quality of neuronal differentiation and reduction in apoptosis that occurred in two-dimensional (2D) and three-dimensional (3D) culture conditions is compared. PC12 and embryonic stem cells are two commonly utilized cell lines for the study of neuronal regeneration. These cells were induced to neuronally differentiate by adding NGF and retinoic acid respectively. Total neurite length and expression of neuronal markers (MAP-2 and β3-tubulin) was assessed by morphometry and immunocytochemistry. Also, TUNEL assay was used to detect apoptosis. Upon exposure to a differentiation media in the 3D fibrin gel, PC12 and embryonic stem cells stopped dividing, had increased adhesion to the substratum, extended neurite processes and expressed neuronal markers. The same results, however, were not observed with the 2D culture. Also, the apoptosis index performed by TUNEL assay demonstrated a reduction in the degree of apoptosis in the 3D culture compared to 2D culture. Fibrin matrix supports growth and neuronal differentiation of PC12 and embryonic stem cells. In addition, the 3D culture enhanced cellular resistance to apoptosis when compared to the 2D culture. It appears as if a 3D culture system may offer a better technique for future neuronal tissue engineering investigations.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 312-316

 

 

Epigallocatechin gallate induces the steady state mRNA levels of pS2 and
PR genes in MCF-7 breast cancer cells

Mohan C Manjegowda, Gauri Deb & Anil M Limaye*

Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati 781 039, India

Received 30 April 2013; Revised 18 December 2013

Investigations using in vitro and in vivo models of breast carcinogenesis have demonstrated anti-neoplastic activity of the green tea polyphenol, epigallocatechin gallate (EGCG). Although a number of molecular targets of EGCG have been identified, its impact on the expression of estrogen target genes is not completely understood. Here, we examined the mRNA expression levels of two estrogen target genes, namely Trefoil Factor 1 (pS2) and Progesterone Receptor (PR) in MCF-7 cells treated with EGCG. We observed that treatment with 40 オM EGCG, which caused only 20% decrease in cell viability, resulted in increased steady state expression levels of pS2 and PR mRNA. This suggests that EGCG may exert its biological activities, at least in part, by influencing the expression of estrogen target genes.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 317-322

 

 

Chronic spinal infusion of loperamide alleviates postsurgical pain in rats

Rakesh Kumar1*, K H Reeta2 & Subrata Basu Ray1

Department of 1Anatomy and 2 Pharmacology

All India Institute of Medical Sciences, New Delhi 110 029, India

Received 8 January 2013; revised 2 December 2013

Plantar incision in rat generates spontaneous pain behaviour. The opioid drug, morphine used to treat postsurgical pain produces tolerance after long-term administration. Loperamide, a potent mu-opioid agonist, has documented analgesic action in various pain conditions. However, loperamide analgesia and associated tolerance following continuous spinal administration in postsurgical pain has not been reported. Chronic spinal infusion of drugs was achieved using intrathecal catheters connected to osmotic minipump. Coinciding with the onset of spinal infusion of loperamide or morphine, rats were subjected to plantar incision. Pain-related behaviour was assessed by Hargreaves apparatus (thermal hyperalgesia) and von Frey filaments (mechanical allodynia). Morphine and loperamide (0.5, 1 and 2 オL/h) induced analgesia was observed until 7th day post-plantar incision in Sprague-Dawley rats. Morphine and loperamide produced dose-dependent analgesia. Loperamide, in the highest dose, produced analgesia till 7th day. However, the highest dose of morphine produced inhibition of thermal hyperalgesia till 5th day and mechanical allodynia only till 3rd day post-plantar incision. Morphine and loperamide produced analgesia in postsurgical pain, which may be mediated through different mechanisms. Longer duration of analgesia with loperamide could probably be due sustained blockade of calcium channels.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 323-331

 

 

Protective effect of Azadirachta indica A. Juss against doxorubicin-induced cardiac toxicity in tumour bearing mice

Ashwani Koul*, Renu Goyal & Sanjay Bharati

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

Received 14 June 2013; revised 13 January 2014

Doxorubicin (DOX) treatment (12 オg/g body weight, once a week for 2 weeks) resulted in a significant decrease in the heart rate along with an increase in QRS, ST, and QT intervals. Histopathological studies showed cardiomyocyte degeneration, cytoplasmic vacuolation and macrophage infiltration in cardiac tissue. A marked increase in the rate of apoptosis was also observed. An increased oxidative stress was evidenced by significantly higher levels of lipid peroxidation (LPO) and depletion of reduced glutathione. A decrease in the activity of cellular antioxidant defence enzymes was also observed. The decrease in the heart rate and ECG alterations were prevented significantly by AAILE (100 オg/g body weight, po) co-treatment, started two weeks prior to DOX treatment and continued till the termination of the experiment. The cardioprotection was also evident from histopathology and decrease in the rate of apoptosis in cardiomyocytes. AAILE co-treatment also prevented DOX-induced increase in LPO and decrease in antioxidant defence enzymes. The results suggest that AAILE administration prevents DOX-induced cardiotoxicity.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 332-343

 

 

Impairment of renal structure and function following heterogeneous chemical mixture exposure in rats

Kiran Morya & Kauresh D Vachhrajani*

Division of Environment and Toxicology, Department of Zoology, Faculty of Science,

The Maharaja Sayajirao University of Baroda, Vadodara 390 002, India

Received 14 January 2013; revised 27 December 2013

Renal structural and functional alterations following an exposure to a heterogeneous chemical mixture (HCM) of phthalic acid di butyl ester, 1, 2謀ichlorobenzene, cadmium chloride and chromium trioxide, administered through oral gavage in low doses (1/100 and 1/1000 of LD50 value of individual chemical) for 60 days, followed by withdrawal till
120 days resulted in significant rise in kidney lipid peroxidation and fall in the activities of enzymatic antioxidants. However, withdrawal of HCM treatment restored most of these altered parameters. Degenerative changes in the kidney included proximal convoluted tubules devoid of brush boarder with cytoplasmic blebbing, dissolution and sloughing of nuclei. Cortical glomeruli were also affected with epithelial disintegration, pyknosis of podocyte nuclei and mesengial cell hyperplasia. The morphological alterations recovered fully in the low dose compared to the high dose treatment group.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 344-351

 

 

Phenotypic plasticity and ecotypic variations in growth and flowering time of Arabidopsis thaliana (L.) under different light and temperature conditions

Shubhangi Moharekar (Lokhande)1,2,*, Sanjay Moharekar1,2, Tsuyoshi Kobayashi1,3, Hiroaki Ishii1,4,
Akihiro Sumida1 & Toshihiko Hara1

1The Institute of Low Temperature Science, Hokkaido University, Sapporo, 060-0819, Japan

Received 12 September 2012; revised 28 October 2013

Four ecotypes of A. thaliana (L.) (Ct-1, Pf-0, Old-1 and Per-1) from low to high latitudes were grown under different light (300 mmol photon m-2s-1 and 150 mmol photon m-2s-1) and temperature (22 and 14 コC) conditions to investigate their effects on phenotypic plasticity and ecotypic variations in plant growth and first flowering time. The results suggest that in A. thaliana low temperature decreases both phenotypic plasticity and ecotypic variations in first flowering time and total dry matter at final harvest under different light intensities. Relative growth rate is the most stable parameter of A. thaliana that is hardly affected by ecotype (no effect), light (no effect) or temperature (small effect) and this may one of the reason why A. thaliana is widely distributed on earth as a result of adaptations to different environments.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 352-358

 

 

Regulation of nitrogen metabolism in salt tolerant and salt sensitive
Frankia strains

 

Amrita Srivastava & Arun Kumar Mishra*

Laboratory of Microbial Genetics, Department of Botany, Banaras Hindu University, Varanasi 221 005, India

Received 23 January 2013; revised 10 October 2013

Effect of salinity (0, 50, 100, 250, 500 and 750 mM NaCl) was observed on some important physiological parameters of nitrogen metabolism such as nitrate uptake, intracellular and extracellular ammonium status and activities of nitrogenase, nitrate reductase, nitrite reductase and glutamine synthetase among Frankia strains differing in their salt tolerance capacity. Nitrogenase activity closely followed the growth pattern with regular decline on NaCl supplementation. All the other enzymes showed optimum activity at 100 mM and declined further. Co-regulation of the nitrate uptake system and sequential enzyme activities plays a crucial role in governing the nitrogen status of strains during salt stress.
HsIi10 experiencing minimum decline in enzyme activities and best possible nitrogen regulation under NaCl replete condition showed adequate nutritional management. Among all the strains, HsIi10 proved to be salt tolerant on account of above features while the salt sensitive strain HsIi8 lacked the ability to regulate various steps of nitrogen metabolism during salinity, and thus Frankia strain HsIi10 can potentially serve as a potential biofertilizer in the saline soil.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 359-368

 

 

Biosynthesis of silver nanoparticles using fresh extracts of Tridax procumbens Linn

Himakshi Bhati訪ushwaha & CP Malik*

School of Life Sciences, Jaipur National University, Jagatpura, Jaipur, India 302 017

Received 12 March 2013; revised 18 November 2013

A simple and eco-friendly method for the synthesis of biogenic nanoparticles (NP痴) using an aqueous solution of
T. procumbens fresh plant extract (leaf and stem) as a bioreductant is reported. The prepared biogenic nanoparticles were well characterized using U.V. visible spectroscopy, scanning electron microscopy, X-ray diffraction and Fourier-transform infrared spectroscopy. The particles were confirmed to be elemental crystal by X-ray diffraction. The potential applications of biosynthesized nanoparticles as antimicrobial (antibacterial and antifungal) against pathogens Escherichia coli, Vibrio cholerae, Aspergillus niger and Aspergillus flavus were demonstrated.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 369-374

 

 

Efficacy of Spodoptera litura multiple nucleopolyhedrovirus after serial passage through the homologous insect larval host

Mudasir Gani*, R K Gupta & K Bali

Division of Entomology, Sher-e-Kashmir University of Agricultural Sciences and Technology, Jammu, Chatha, India 180 009

Received 9 August 2013; revised 28 October 2013

An originally isolated baculovirus, Spodoptera litura multiple nucleopolyhedrovirus (SpltMNPV) was serially passed through the S. litura larvae for upto four generations to determine the mean number of occlusion bodies (OBs) harvested per larva and their efficacy in terms of infectivity, feeding cessation and speed of kill of host larvae. The results revealed that the mean number of OBs harvested per larva increased significantly with increase in the dose of SpltMNPV at each passage and the yield was significantly lower in original stock wild-type SpltMNPV (P0) as compared to serially passed SpltMNPV (P1, P2, P3 and P4). Laboratory bioassays indicate that median lethal doses (LD50), median times to feeding cessation (FT50) and median survival times (ST50) of P0, P1, P2, P3 and P4 were significantly different from each other. The OBs of each passage when tested for their cross-infectivity against Spodoptera exigua and Spilarctia obliqua revealed significant reduction in their mortality. These results indicate that serially passed SpltMNPV is more host specific and more effective biocontrol agent than the original stock wild-type virus and can be adopted for mass production as a viral pesticide for control of the S. litura.

 

 

Indian Journal of Experimental Biology

Vol. 52, April 2014, pp. 375-382

 

 

Pharmacophore based approach to design inhibitors in Crustaceans: An insight into the molt inhibition response to the receptor guanylyl cyclase

Sajal Shrivastava & S Adline Princy*

Quorum Sensing Laboratory, SASTRA痴 Hub for Research and Innovation, SASTRA University, Thanjavur 613 401, India

Received 13 May 2013; revised 8 January 2014

The first set of competitive inhibitors of molt inhibiting hormone (MIH) has been developed using the effective approaches such as Hip-Hop, virtual screening and manual alterations. Moreover, the conserved residues at 71 and 72 positions in the molt inhibiting hormone is known to be significant for selective inhibition of ecdysteroidogenesis; thus, the information from mutation and solution structure were used to generate common pharmacophore features. The geometry of the final six-feature pharmacophore was also found to be consistent with the homology-modeled MIH structures from various other decapod crustaceans. The Hypo-1, comprising six features hypothesis was carefully selected as a best pharmacophore model for virtual screening created on the basis of rank score and cluster processes. The hypothesis was validated and the database was virtually screened using this 3D query and the compounds were then manually altered to enhance the fit value. The hits obtained were further filtered for drug-likeness, which is expressed as physicochemical properties that contribute to favorable ADME/Tox profiles to eliminate the molecules exhibit toxicity and poor pharmacokinetics. In conclusion, the higher fit values of CI-1 (4.6), CI-4 (4.9) and CI-7 (4.2) in conjunction with better pharmacokinetic profile made these molecules practically helpful tool to increase production by accelerating molt in crustaceans. The use of feeding sub-therapeutic dosages of these growth enhancers can be very effectively implemented and certainly turn out to be a vital part of emerging nutritional strategies for economically important crustacean livestock.