Indian Journal of Experimental Biology

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VOLUME 51

NUMBER 6

JUNE 2013

CODEN: IJEB (A6) 51 (6) 407-480 (2013)

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

 

CONTENTS

 

 

Papers

 

 

 

Isolation, purification and characterization of the egg-yolk proteins from the oocytes of the Indian freshwater murrel, Channa punctatus (Bloch)

411

 

 

Om Prakash, N Sehgal, K V Rani & N Aggarwal

 

 

 

The lld mutation in Pisum sativum used as a genetic tool to discern the plant leaflet/leaf developmental process

421

 

 

Vishakha Sharma, Bhumi Nath Tripathi & Sushil Kumar

 

 

 

Anti-depressant like activity of N-n-butyl-3-methoxyquinoxaline-2-carboxamide (6o) a 5-HT3 receptor antagonist

435

 

 

Shvetank Bhatt, Radhakrishnan Mahesh, Thangaraj Devadoss & Ankur Jindal

 

 

 

Anxiolytic-like effect of etazolate, a type 4 phosphodiesterase inhibitor in experimental models of anxiety

444

 

 

Ankur Jindal, Radhakrishan Mahesh & Shvetank Bhatt

 

 

 

Anxiolytic and anticonvulsant activity of methanol extract of leaves of Alternanthera brasiliana (L.) Kuntze (Amaranthaceae) in laboratory animals

450

 

 

Chandana C Barua, Shameem A Begum, Acheenta G Barua, Rumi S Borah & Mangala Lahkar

 

 

 

Hypolipidemic effect of a novel biflavonoid from shells of Camellia oleifera (Abel.)

458

 

 

Y Ye, H T Xing & Y Guo

 

 

Effect of Aegle marmelos on alloxan induced early stage diabetic nephropathy in rats

464

 

 

Rajbir Bhatti, Shikha Sharma, Jatinder Singh, Amarjit Singh & M P S Ishar

 

 

 

In planta detection of Xanthomonas axonopodis pv. commiphorae using fyuA and rpoD genes

470

 

 

Jatindra Nath Samanta & Kunal Mandal

 

 

 

Effects of low concentrations of a polychlorinated biphenyl, Aroclor 1254 on membrane bound ion dependent ATPases in mice liver

477

 

 

Shweta Pathak & Rahul Kundu

 

 

Announcement

 

International Conference on Advances in Biotechnology and Bioinformatics (ICABB 2013)

25­27 November 2013, Pune, India

 

Jointly organised by Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth (Deemed University), Pune and The Biotech Research Society, India, the International Conference on Advances in Biotechnology and Bioinformatics (ICABB 2013) and X convention of Biotech Research Society India, will be held at Pune, India. The scientific program of the conference would comprise plenary and invited lectures from the internationally acclaimed and eminent experts. The theme areas are: (i) Plant Biotechnology, (ii) Microbial and Industrial Biotechnology, (iii) Animal and Medical Biotechnology, (iv) Nano-Biotechnology, (v) Systems Biology and Bioinformatics and (vi) Biotechnology, Business and IPR. For details, please contact Dr Neelu Nawani, Organising Secretary, ICABB 2013, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth (Deemed University), Survey No. 87/88, Mumbai-Bangalore Expressway, Tathawade, Pune 411 044, India; Mobile: 91-9822648395, Telephone: 91-20-65101870, E-mail: icabb2013@dpu.edu.in, or the General Chair, Prof. Ashok Pandey, Centre for Biofuels and Biotechnology Division, CSIR-National Institute for Interdisciplinary Science and Technology (NIIST), Trivandrum, 695 019, India; Telephone: 91-471-2515279, Fax: 91-471-2491712, E-mail: pandey@niist.res.in; ashokpandey56@yahoo.co.in, or visit the website: www.icabb2013.dpu.edu.in

 

_______________________

 

Editor’s 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

 

 

 

Author Index

Aggarwal N

411

 

 

Barua Acheenta G

450

Barua Chandana C

450

Begum Shameem A

450

Bhatt Shvetank

435,444

Bhatti Rajbir

464

Borah Rumi S

450

 

 

Devadoss Thangaraj

435

 

 

Guo Y

458

 

 

Ishar M P S

458

 

 

Jindal Ankur

435,444

 

 

Kundu Rahul

477

 

 

Lahkar Mangala

450

 

 

Mahesh Radhakrishnan

435,444

Mandal Kunal

470

 

 

Om Prakash

411

 

 

Pathak Shweta

477

 

 

Rani K V

411

 

 

Samanta Jatindra Nath

470

Sehgal N

411

Sharma Shikha

458

Sharma Vishakha

421

Singh Amarjit

458

Singh Jatinder

458

Sushil Kumar

421

 

 

Tripathi Bhumi Nath

421

 

 

Xing H T

458

 

 

Ye Y

458

 

Keyword Index

5-HT3 receptor antagonists

435

 

 

Acropetal movement

470

Adaxial-abaxial differentiation

421

Aegle marmelos

464

Alloxan

464

Alternanthera brasiliana

450

Anti-anxiety

444

Anticonvulsant activity

450

Antioxidant

464

Antioxidative activity

458

Aroclor 1254

477

ATPases

477

 

 

Biflavonoid

458

 

 

Camellia oleifera

458

Channa punctatus

411

Cup shaped leaf

421

Diabetic nephropathy

464

 

 

Egg-yolk extract

411

Elevated plus maze

450

Etazolate

444

Extracts

458

 

 

Head twitches

435

Hole board

450

Hypolipidemic effect

458

 

 

Leaf development mechanism

421

Leaf vein network

421

Light/dark exploration

450

Lipovitellin

411

Liver

477

Locomotor activity

450

 

 

Mediolateral polarity

421

Mice

477

Molecular detection

470

Murrel

411

 

 

Olfactory bulbectomy

435

Open field test

450

 

 

Phosphodiesterase 4

444

Phosvitin

411

Proximodistal growth

421

 

 

Reserpine induced hypothermia

435

 

 

Seizures

450

 

 

Tail suspension test

435

 

 

Xanthomonas axonopodis pv. commiphorae (Xac)


470

 

 

 

            Correspondent author is marked by *

 

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 411-420

 

 

Isolation, purification and characterization of the egg-yolk proteins from the oocytes of the Indian freshwater murrel, Channa punctatus (Bloch)

Om Prakasha, N Sehgalb, K V Ranib & N Aggarwalb*

aDepartment of Zoology, SriVenkateswara College, University of Delhi, New Delhi 110 021, India

bDepartment of Zoology, University of Delhi, Delhi 110007, India

Received 19 September 2012; revised 8 March 2013

In oviparous organisms, yolk accumulation in the oocytes is critical and indispensable for the development of the newly hatched young ones. In fish and many other oviparous vertebrates, the major constituents of the egg-yolk are synthesized as a precursor in the liver. The precursor is transported to the oocyte for uptake and cleaved into major yolk proteins lipovitellin, phosvitin and β’-components. The eggs of Channa punctatus are pelagic, have large oil globule and exceptionally high lipid content. Lipovitellin was isolated by single step gel filtration chromatography on Sepharose 6B. Purified native lipovitellin showed immunological reactivity with vitellogenin antiserum. Phosvitin isolated by phenol extraction method could not be visualized with routine protein staining methods, whereas incorporation of trivalent ions in the coomassie brilliant blue stained phosvitin. It was characterized by in vivo labeling of egg-yolk proteins with 32P. The molecular mass of murrel phosvitin was less than 14,000 kDa.

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 421-434

 

 

 

The lld mutation in Pisum sativum used as a genetic tool to discern
the plant leaflet/leaf developmental process

Vishakha Sharma1,2,3, Bhumi Nath Tripathi2 & Sushil Kumar1, 3*

1National Institute of Plant Genome Research (NIPGR), Aruna Asaf Ali Marg, New Delhi 110 067, India

2Banasthali University, PO Banasthali Vidhyapeeth 304 022, India

3SKA Institution for Research, Education and Development (SKAIRED), 4/11 Sarv Priya Vihar, New Delhi 110 016, India

Received 8 October 2012

Leaves of P. sativum the double mutant genotype tendril-less (tl) leaflet-development (lld), due to the action of lld mutation, produce many leaflets that are aborted at different stages of development. Morphological, vein pattern and histological observations showed that aborted leaflets became cup/bell/trumpet (cup) shaped because of segmental differentiation in the leaflet primordium. Cup’s inside lamina surface was adaxial and outer surfaces of cup and its stem were abaxial. The lld cups were phenotypically homologous to aborted leaves described in Arabidopsis thaliana mutants, angustifolia and those which underexpressed the HD-ZIP III proteins. Leaflet primordium was found to grow and establish three dimensional polarities apex-downwards. Primordium produced lateral outgrowth on one side of midvein. Differentiation, in the outgrowth, of secondary veins, whose xylem tissues faced each other, established the adaxial-abaxial polarity. Lateral outgrowth then developed a cavity which got bounded by future adaxial epidermis. Further growth, veinlet formation, differentiation of palisade parenchyma and spongy parenchyma followed. Opening of lateral outgrowth at its outer midline produced a flat leaflet with lateral lamina spans. The structural and functional correspondence between leaflet and simple leaves suggested commonality between leaf and leaflet development mechanisms. A molecular model for the lld led leaflet abortion was also provided.

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 435-443

 

 

Anti-depressant like activity of N-n-butyl-3-methoxyquinoxaline-2-carboxamide (6o) a 5-HT3 receptor antagonist

Shvetank Bhatt*, Radhakrishnan Mahesh, Thangaraj Devadoss & Ankur Jindal

Department of Pharmacy, Birla Institute of Technology & Science, Pilani 333 031, India

Received 10 September 2012; revised 14 March 2013

The compound 6o (at 0.5, 1 and 2 mg/kg, ip) with optimum log P and pA2 value, was subjected to forced swim test (FST) and tail suspension test (TST). The compound 6o significantly reduced the duration of immobility in mice without affecting the base line locomotion in actophotometer. Moreover, 6o (2 mg/kg, ip), potentiated the 5-hydroxytryptophan
(5-HTP)-induced head twitch responses in mice and at 1 and 2 mg/kg, ip antagonized the reserpine-induced hypothermia (RIH) in rats. In interaction studies with various standard drugs/ligands using FST, 6o (1 and 2 mg/kg, ip) potentiated the anti-depressant effect fluoxetine (5 mg/kg, ip) and reversed the depressant effect of parthenolide (1 mg/kg, ip) by reducing the duration of immobility. Furthermore, 6o (1 and 2 mg/kg, ip) potentiated the effect of bupropion (10 mg/kg, ip) in TST. The behavioural anomalies of the olfactory bulbectomised (OBX) rats were augmented by chronic 6o (1 and 2 mg/kg) treatment as observed from the modified open field test (parameters: ambulation, rearing, fecal pellet). The results suggest that compound 6o exhibited anti-depressant like effect in rodent models of depression.

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 444-449

 

 

Anxiolytic-like effect of etazolate, a type 4 phosphodiesterase inhibitor
in experimental models of anxiety

Ankur Jindal*, Radhakrishan Mahesh & Shvetank Bhatt

Department of Pharmacy, Birla Institute of Technology & Science, Pilani 333 031, India

Received 17 September 2012; revised 14 March 2013

Etazolate is a selective inhibitor of type 4 phosphodiesterase (PDE4) class enzyme. Antidepressant-like effect of etazolate has been previously demonstrated in the rodent models of depression. The present study was designed to investigate the anxiolytic-like activity of etazolate in experimental mouse models of anxiety. The putative anxiolytic effect of etazolate (0.25-1 mg/kg, ip) was studied in mice by using a battery of behavioural tests of anxiety such as elevated plus maze (EPM), light/dark (L/D) aversion, hole board (HB) and open field (OFT) with diazepam (2 mg/kg, ip) as reference anxiolytic. Like diazepam (2 mg/kg, ip), etazolate (0.5 and 1 mg/kg, ip) significantly increased the percentage of both time spent and entries into open arms in the EPM test. In the L/D test etazolate (0.5 and 1 mg/kg, ip) increased the both total time spent in and latency time to leave the light compartment. Etazolate (0.5 and 1 mg/kg, ip) also significantly increased head dipping scores and time spent in head dipping, whereas significantly decreased the head dipping latency in HB test. In addition, etazolate (0.5 and 1 mg/kg, ip) significantly increased the ambulation scores (square crossed) and number of rearing in OFT. In conclusion, these findings indicated that etazolate exhibited an anxiolytic-like effect in experimental models of anxiety and may be considered an alternative approach for the management of anxiety disorder.

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 450-457

 

 

Anxiolytic and anticonvulsant activity of methanol extract of leaves of Alternanthera brasiliana (L.) Kuntze (Amaranthaceae) in laboratory animals

Chandana C Barua1*, Shameem A Begum2, Acheenta G Barua3, Rumi S Borah4 & Mangala Lahkar5

Department of 1Veterinary Pharmacology and Toxicology, 2Veterinary Pathology, 3Veterinary Public Health & Hygiene and
4Animal Production and Management, College of Veterinary Science, Assam Agricultural University, Khanapara, Guwahati 781 022, India and 5Department of Pharmacology, GMCH, Guwahati 781 006, India

Received 9 October 2012; revised 13 March 2013

Anxiety related disorders are the most common mental illnesses and major cause of disability in man. Anxiolytic activity of methanol extract of leaves of A. brasiliana (L.) Kuntze (MEAB) was evaluated using hole board (HB), open field (OF), elevated plus maze (EPM) and light/dark exploration test (LDE) in mice. Its locomotor activity was studied using actophotometer and anticonvulsant effect was studied using maximal electroshock-induced seizures and pentylenetetrazole-induced seizures in mice. Single oral administration of MEAB at different doses (100, 300 and 600 mg/kg, ip) significantly increased the number and duration of head poking in the HB test; rearing, assisted rearing and number of square traveled in the OF test; entries and time spent in open arm in the EPM test; time spent in lighted box, and numbers of crossings and transfer latency time in the LDE test. There was significant reduction in the time spent in close arm in the EPM test and time spent in dark box in LDE test. In the actophotometer, the activity count was reduced in MEAB and diazepam treated group than control group. All the three doses of the extract significantly reduced the duration of seizures induced by pentylenetetrazole (chemoshock convulsion). However, the extract did not show any appreciable effect in electroshock convulsion model. The results of the present study suggest promising anxiolytic and anticonvulsant activity of MEAB which might be accredited to different phytoconstituents like alkaloids, steroids and triterpenes present in the methanol extract of A. brasiliana.

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 458-463

 

 

Hypolipidemic effect of a novel biflavonoid from shells of Camellia oleifera (Abel.)

Y Ye*, H T Xing & Y Guo

Pharmaceutical Engineering Department, School of Chemistry and Chemical Engineering, South
China University of Technology, Guangzhou 510 640, PR China

Received 22 October 2012

Camellia oleifera Abel. [C. oleosa (Lour.) Rehd.], an evergreen plant, is used for healthful oil production, but the shells are always discarded and need to be utilized. The present study was undertaken to explore the effect of extracts from the shells of C. oleifera on adjusting cardiovascular system. A flavonoid was obtained by reflux extraction of the shells in 70% methanol, hydrolysis in 2 M hydrochloric acid, and crystallization in acetone. Its structure was identified as a novel biflavonoid. Mice model of hyperlipidemia was setup by high fat diet for 30 d to evaluate the hypolipidemic effect of the biflavonoid at dose of 50, 100 and 200 mg/kg/d (ig). Antioxidative activity was determined by levels of malondialdehyde (MDA), superoxidase dismutase (SOD) and glutathione peroxidase (GSH-Px) in mice serum. The biflavonoid significantly controlled mice weight and liver coefficient, decreased the content of total cholesterol and triglyceride, promoted the level of high density lipoprotein in a dose dependent manner. The significant decrease of MDA content and increase of SOD and GSH-Px activity indicated it enhanced antioxidative capacity in vivo and was ascribed to hypolipidemic effect. The biflavonoid is useful in the prevention of high fat diet induced hyperlipidemia.

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 464-469

 

 

Effect of Aegle marmelos (L.) Correa on alloxan induced early stage diabetic
nephropathy in rats

Rajbir Bhatti1*, Shikha Sharma1, Jatinder Singh2, Amarjit Singh2 & M P S Ishar1,3

1 Department of Pharmaceutical Sciences, Guru Nanak Dev University, Amritsar, India.

2 Department of Pharmacology, Government Medical College, Amritsar, India

Received 31 July 2012; revised 28 January 2013

Diabetic nephropathy (DN) has a complex pathogenesis and poor prognosis due to the lack of therapeutic interventions. The present study investigates the effect of A. marmelos leaf extract (AME) on early alloxan induced DN. The treatment with AME was found to significantly decrease the fasting blood sugar, total cholesterol, blood urea, creatinine and renal TBARS and increased the levels of renal reduced glutathione and catalase significantly as compared to the diabetic control group. The maximum dose-dependent protection was observed at a dose of 200 mg kg-1. Histological examination revealed marked reversal of the morphological derangements with AME treatment as indicated by a decrease in glomerular expansion, tubular dilatation and inflammatory cells. The present results conclude that AME treatment has a significant ameliorative effect on early changes induced in the kidneys by alloxan and improves the outcome of DN.

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 470-476

 

 

In planta detection of Xanthomonas axonopodis pv. commiphorae using
fyuA and rpoD genes

Jatindra Nath Samantaa* & Kunal Mandalb

aDirectorate of Medicinal and Aromatic Plants Research (ICAR), Boriavi, Anand 387 310, India

bDirectorate of Mushroom Research, (ICAR), Chambaghat, Solan 173 213, India

Received; 26 December 2012

Guggal is tapped for extraction of medicinally important oleo–gum–resin (guggul) by inoculating the stem bark with natural gum suspension containing pathogenic bacterium Xanthomonas axonopodis pv. commiphorae (Xac). The tree dies in the process. In absence of any specific medium for isolation of Xac, it is difficult to assess spread of the pathogen within the plant. A PCR based molecular detection technique using fyuA and rpoD gene specific primers is described here. The primers amplified products only from Xac and not from host tissues or common saprophytes. The method was sensitive enough to produce positive signals for up to 4.4 bacterial cells or 2 pg target DNA per reaction mixture. However, PCR inhibitors present in plant tissues drastically reduced the limit of detection. A simple overnight incubation of surface sterilised plant tissues in nutrient medium was introduced to increase pathogen titre and to overcome this problem. This technique was successfully used to measure spread of Xac in plant tissues away from the site of inoculation. The pathogen showed preference for acropetal movement and did not spread to 7–8 cm below the site of inoculation till 15 days after inoculation. This suggests possibility to manage the disease through plant surgery.

 

 

Indian Journal of Experimental Biology

Vol. 51, June 2013, pp. 477-480

 

 

Effects of low concentrations of a polychlorinated biphenyl, Aroclor 1254 on membrane bound ion dependent ATPases in mice liver

Shweta Pathak & Rahul Kundu*

Department of Biosciences, Saurashtra University, Rajkot 360 005, India

Received 2 November 2012; revised 8 March 2013

Aroclor 1254, a polychlorinated biphenyl, is present in the environment in low concentration but references on its toxic effects on liver cell membrane proteins and the mechanism of actions are not abundantly available. Therefore, the present study was undertaken to investigate the low level, sub-acute dose and exposure duration dependent effects of Aroclor 1254 on total, Na+, K+, Ca2+ and Mg2+-ATPases of the mouse liver. The hypotheses tested in the present study were, (a) whether the low, environmentally available dose and the exposure durations of Aroclor 1254 affects the membrane-bound ion dependent ATPases, and (b) if a response was observed, whether it is a direct or indirect effects of the toxicant. Groups of mice were exposed to different doses (0.1 and 1mg kg-1 body weight d-1) and exposure durations (4 d, 8 d and 12 d) of Aroclor 1254. The results indicated significant exposure duration dependent changes in the specific activity of the selected membrane bound ATPases. As the observed changes were mostly enzyme stimulation after toxication through oral administration, the effects of the Aroclor were possibly indirect, through complex chain of reactions.