Indian Journal of Experimental Biology

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VOLUME 51

NUMBER 5

MAY 2013

CODEN: IJEB (A6) 51 (5) 343-406 (2013)

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

 

CONTENTS

 

 

Papers

 

 

 

Purification of heparin binding oviduct specific proteins and its effect on in vitro embryo development in cattle

347

 

 

Aditya K Sharma, Sushil K Mohapatra, A K Mohanty & S K Das

 

 

 

Cloning and expression of Bos indicus interleukin-4 in mammalian cells

352

 

 

Prashanth T, G R Reddy, V V S Suryanaryana & H J Dechamma

 

 

 

Effect of epigallocatechin-3-gallate on inflammatory mediators release in LPS-induced Parkinson's disease in rats

357

 

 

Jehan S AL-amri, Magda M Hagras & Mohamed I Mujallid

 

 

 

Antidiabetic activity of heart wood of Pterocarpus marsupium Roxb. and analysis of phytoconstituents

363

 

 

Akansha Mishra, Rohit Srivastava, Swayam Prakash Srivastava, Sudeep Gautam, Akhilesh Kumar Tamrakar, Rakesh Maurya & Arvind K Srivastava

 

 

 

Role of tricalcium phosphate implant in bridging the large osteoperiosteal gaps in rabbits

375

 

 

Satish Chandra Goel, Davinder Singh, Amit Rastogi, Vinay Kumaraswamy, Ashank Gupta & Nitin Sharma

 

 

Sub-acute toxicity of cultured mycelia of Himalayan entomogenous fungus Cordyceps sinensis (Berk.) Sacc. in rats

381

 

 

Harsahay Meena, Kshetra Pal Singh, Prem Sing Negi & Zakwan Ahmed

 

 

 

Effect of UV-C on thylakoid arrangement, pigment content and nitrogenase activity in the cyanobacterium Microchaete sp.

388

 

 

J K Sahu & M imek

 

 

 

Microalgae mediated synthesis of silver nanoparticles and their antibacterial activity against pathogenic bacteria

393

 

 

S S Sudha, Karthic Rajamanickam & J Rengaramanujam

 

 

 

A multilayered supramolecular self-assembed structures from soybean oil by in situ polymerization and its applications

400

 

 

Varadharajan Kavitha & Arumugam Gnanamani

 

 

 

Letter to the Editor

 

 

 

Protective effects of subchronic caffeine administration on cisplatin induced urogenital toxicity in male mice

406

 

 

Hamid Nasri

 

 

 

_______________________

 

Announcements

 

Ist International and IIIrd National Conference on Biotechnology, Bioinformatics and Bioengineering

28 and 29 June 2013, Hotel Udayee International, Tirupati, India

 

Organized by the Society for Applied Biotechnology (www.sabt.org.in), the topics to be covered in the Conference include, but are not limited to: (1) Biodiversity and Biotechnology, (2) Plant, Food and Agricultural Biotechnology, (3) Animal, Marine and Fisheries Biotechnology, (4) Environmental and Industrial Biotechnology, (5) Pharmaceutical and Medical Biotechnology, (6) Microbial and Analytical Biotechnology, (7) Nanobiotechnology and Nanobioengineering, (8) Genomics, Proteomics and Bioinformatics, (9) Enzyme, Protein and Tissue Engineering, (10) Biochemical and Biomedical Engineering, and (11) Biotechnology Research and Opportunities. For further details, please contact: Dr. Devarajan Thangadurai, Department of Botany, Karnatak University, Dharwad, 580 003, India. Mobile: +91 9482685270/9482685280; E-mail: drthangaduraid@gmail.com; thanga@sabt.org.in or visit website: www.sabt.org.in/conferences.htm

 

 

_______________________

 

 

3rd Annual Conference TSICON 2013 and Ist International Conference on Biology of Natural Toxins

19-21 December 2013, BITS Pilani, K K Birla Goa Campus Goa, India

 

The conference aims to provide a common platform for all researchers, clinicians, students, conservationists and industries working on different aspects of natural toxins of animals, microbes and plant, snake bite management and related environmental issues to discuss their research findings. The conference will consist of plenary sessions, invited lectures, oral and poster presentations. Scientific areas to be covered are (i) Venomics, (ii) Clinical toxinology, (iii) Characterization of natural toxins, (iv) Pharmacology, pharmacokinetics and drug discovery, (v) Anti-venom/Antidotes and Industry, (vi) Ethics in toxin research, and (vii) Conservation of poisnous and venomous species. For details please contact Dr. Dibakar Chakrabarty, Department of Biological Sciences, BITS Pilani K.K Birla Goa Campus, Goa 403 726. Telephone: +91 832 2580173; E-mail: tsicon2013@gmail.com or visit website: http://www.tsicon2013.org/; or Prof. Antony Gomes, Laboratory of Toxinology and Experimental Pharmacodynamics, Department of Physiology, University of Calcutta, 92, A. P. C. Road, Kolkata 700 009. Mobile: +91-9433139031 Fax: 91-033-2351975; E-mail: secretary.tsi@gmail.com

_______________________

 

 

Author Index

 

Ahmed Zakwan

381

AL-amri Jehan S

357

 

 

Das S K

347

Dechamma H J

352

 

 

Gautam Sudeep

363

Gnanamani Arumugam

400

Goel Satish Chandra

375

Gupta Ashank

375

 

 

Hagras Magda M

357

 

 

Kavitha Varadharajan

400

Kumaraswamy Vinay

375

 

 

Maurya Rakesh

363

Meena Harsahay

381

Mishra Akansha

363

Mohanty A K

347

Mohapatra, Sushil K

347

Mujallid Mohamed I

357

 

 

Nasri Hamid

406

Negi Prem Sing

381

 

 

Rajamanickam Karthic

393

Rastogi Amit

375

Reddy G R

352

Rengaramanujam J

393

 

 

Sahu J K

388

Sharma Aditya K

347

Sharma Nitin

375

imek M

388

Singh Davinder

375

Singh Kshetra Pal

381

Srivastava Arvind K

363

Srivastava Rohit

363

Srivastava Swayam Prakash

363

Sudha S S

393

Suryanaryana V V S

352

 

 

T Prashanth

352

Tamrkar Akhilesh Kumar

363

 

 

Keyword Index

 

Acetylene reduction assay

388

Antidyslipidemic effect

363

Antihyperglycemic effect

363

Antimicrobial agents

393

 

 

Biopolymer

400

 

 

Carotenoid

388

Cattle

347

Ceramic

375

Chinese hamster ovary cells

352

Chlorophyll-a

388

Cordyceps sinensis

381

Cyanobacteria

393

Cytokines

352

 

 

Dendritic cells

352

 

 

Embryo

347

Epigallocatechin-3-gallate

357

Glucose uptake

363

 

 

High fat diet

363

 

 

In vitro fertilization

347

Interleukin-4

352

 

 

Lipopolysaccharide

357

 

 

Microchaete

388

Microcoleus sp

393

Multilayered structures

400

 

 

Nitric oxide

352

Nitric oxide

357

 

 

Osteoconduction

375

Osteoperiosteal gap

375

Oviduct specific proteins

347

Parkinson's disease

357

Polymerization

400

Pterocarpus marsupium
heart wood


363

 

 

Resistance

393

 

 

Self-assembly

400

Silver nanoparticles

393

Skeletal muscle cells

363

Soybean oil

400

Sub-acute toxicity

381

Supramolecular structures

400

 

 

TNF-a

357

Tricalcium phosphate

375

 

 

Ultraviolet-C

388

 

 

Correspondent author is marked by *

 

 

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 347-351

 

 

Purification of heparin binding oviduct specific proteins and its effect on in vitro embryo development in cattle

Aditya K Sharma1, Sushil K Mohapatra2, A K Mohanty2 & S K Das1*

1Dairy Biotechnology Laboratory, Eastern Regional Station, National Dairy Research Institute, Kalyani 741 235, India

2Animal Biotechnology Centre, National Dairy Research Institute, Karnal 132 001, India

Received 7 September 2012; revised 27 December 2012

The objective of this study was to see the effect of purified heparin binding oviduct specific proteins (OSP) as media supplement on in vitro embryo developmental competence in cattle. The oviduct specific proteins were isolated from abattoir cattle oviducts and precipitated, dialyzed and at the end purified by high performance liquid chromatography system. The SDS-PAGE profile of eluted heparin binding protein (HBP) fraction showed bands between ~66 - ~97 kDa, while heparin unbinding protein (HUBP) fraction showed two bands at ~66 kDa and in total protein (TP) bands were ~60 - ~95 kDa. Collected all three OSP fractions were used as a media supplement in three different concentrations (0, 5 and 20 g/mL) for in vitro maturation of immature oocytes, in vitro fertilization and culture of presumptive embryos at 38.5 C in 5% CO2 incubator with maximum humidity. The highest cleavage rate (73.402.36%) was observed at 5 g/mL concentration level and lowest cleavage rate (27.631.89%) was obtained in 20 μg/mL total protein (TP) fraction. The highest blastocyst formation (26.471.47%) also occurred in 5 g/mL concentration of total protein (TP) fraction and the lowest blastocyst rate (3.601.80%) was achieved at 20 g/mL HBP fraction. The highest cleavage rate in the control group was 60.452.66% in TP fraction and blastocyst formation was 11.662.54% in HUBP fraction which was not significantly differ from HBP fraction. These results indicate that at 5 g/mL of total OSP fraction (TP) and HBP used as media supplement increased the cleavage rate significantly as compared to HUBP fraction, and total OSP fraction (TP) increased blastocyst formation significantly (P<0.05) as compared to HBP & HUBP fraction.

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 352-356

 

 

Cloning and expression of Bos indicus interleukin-4 in mammalian cells

Prashanth Ta, G R Reddya, V V S Suryanaryanaa & H J Dechammaa,*

aFoot and Mouth Disease Research Laboratory, Indian Veterinary Research Institute, Bangalore Campus, Hebbal,
Bangalore 560 024, India

Received 4 October 2012; revised 25 February 2013

Dendritic cells (DC) which are located at the interface of innate and adaptive immunity are targets of infection by many RNA and DNA viruses. Advances in the ex vivo generation of monocyte derived non proliferating dendritic cells have been used for clinical application like immunotherapy. IL-4 cytokine plays essential role in the maturation and generation of DCs. Bos indicus interleukin 4 (boIL-4) 408 bp was amplified from PBMCs and cloned in pBSIIKS+ vector. The sequence analysis showed N terminal 69 bp signal sequence and one N-glycosylation site. The phylogenetic tree analysis showed that Bos indicus IL-4 is closely related to the ruminant IL-4 and least sharing of genetic line of human and mouse IL-4. The recombinant boIL-4 protein was expressed in CHO cells which secreted a 16 kDa protein which was confirmed by SDS PAGE and western blotting. The rec-boIL-4 protein proliferated the bovine PBMCs, decreased production of nitric oxide in antigen stimulated macrophages, and phagocytosed the micro particles confirming its activity on dendritic cells.

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 357-362

 

 

Effect of epigallocatechin-3-gallate on inflammatory mediators release in LPS-induced Parkinson's disease in rats

Jehan S AL-amri1, Magda M. Hagras2,3,* & Mohamed I. Mujallid1

1Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia

2Department of Pharmacology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia

3Department of Pharmacology, Faculty of Medicine, Suez Canal University, Cairo, Egypt

Received 13 July 2012; revised 5 February 2013

Degeneration of dopamine (DA)-containing neurons in the substantia nigra of the midbrain causes Parkinson's disease (PD). Although neuroinflammatory response of the brain has long been speculated to play a role in the pathogenesis of this neurological disorder, the mechanism is still poorly understood. The aim of the present study was to examine the effect of epigallocatechin-3-gallate (EGCG) in prevention of inflammatory mediators release and protection of dopaminergic neurons from lipopolysaccharide (LPS)-induced neurotoxicity. A single intraperitoneal injection of LPS (15 mg/kg) in male Sprague Dawley rats resulted in an increase of midbrain content of TNF-a, NO and a decrease of DA level at 4, 24 h, 3 and 7 days compared to the control. In addition, LPS reduced the number and the density of tyrosine hydroxylase-immunoreactive (TH-ir) neurons in the midbrain at 7 days. Pretreatment with EGCG (10 mg/kg) 24 h before LPS for 7 days decreased TNF-a and NO compared to LPS-treated rats. Moreover, it increased DA level and preserved the number and the density of TH-ir neurons compared to LPS group. In conclusion, EGCG was found to have a potential therapeutic effect against LPS-induced neurotoxicity via reducing TNF-a and NO inflammatory mediators and preserving DA level in midbrain.

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 363-374

 

 

Antidiabetic activity of heart wood of Pterocarpus marsupium Roxb. and
analysis of phytoconstituents

Akansha Mishra1, Rohit Srivastava1, Swayam Prakash Srivastava1, Sudeep Gautam1, Akhilesh Kumar Tamrakar1,
Rakesh Maurya2 & Arvind K Srivastava1*

Divisions of 1Biochemistry and 2Medicinal and Process Chemistry

CSIR-Central Drug Research Institute (CDRI), Lucknow 226001, India

Received 10 April 2012; revised 25 February 2013

The crude powder, ethanolic extract and aqueous, chloroform, hexane and n-butanol soluble fractions of ethanolic extract of heart wood of P. marsupium showed marked improvement on oral glucose tolerance post sucrose load in normal rats. All these fractions except aqueous fraction showed improvement on oral glucose tolerance post sucrose load on streptozotocin (STZ)-induced diabetic rats. The crude powder, ethanolic extract and hexane and n-butanol fractions showed marked decline in blood glucose level on STZ-induced diabetic rats. The ethanolic extract (100 mg/kg body weight) when given to STZ-induced diabetic rats for 10 consecutive days declined blood glucose, improved OGTT and increased their serum insulin levels. The ethanolic extract also showed marked improvement on oral glucose tolerance on high fat-low dosed STZ-induced diabetic rats and neonatally STZ treated rats. The ethanolic extract of P. marsupium also showed marked antidyslipidemic effects on high fat diet fed Syrian golden hamsters. Altered renal and hepatic function markers and serum insulin levels of high fat diet fed-low dosed STZ-treated diabetic rats were also found towards normalization when these animals were treated with ethanolic extract of P. marsupium for 28 consecutive days. The four out of five phenolic C-glycosides isolated from n-butanol fraction of ethanolic extract of P. marsupium enhanced glucose uptake by skeletal muscle cells (C2C12) in a dose dependent manner. It may primarily be concluded that phenolic-C-glycosides present in P. marsupium heart wood are the phytoconstituents responsible for the antihyperglycemic activity and validate the claim of antidiabetic activity of heart wood of P. marsupium.

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 375-380

 

 

Role of tricalcium phosphate implant in bridging the large osteoperiosteal
gaps in rabbits

Satish Chandra Goel*, Davinder Singh, Amit Rastogi, Vinay Kumaraswamy, Ashank Gupta & Nitin Sharma

Department of Orthopaedics, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221 005, India

Received 22 May 2012; revised 18 February 2013

Treatment options for large osteoperiosteal defects are limited and that which are available are not ideal. Osteoperiosteal defect were created in ulnae of both forelimbs of rabbits and tricalcium phosphate implant was used to bridge the gap. Amongst the 35 implanted ulnae, one implant got dislodged. Rest of the implants showed good adherence to host bone until the final follow up. Five control rabbit limbs (in which no implants were put) showed persistent bone gap. Histological and Electron microscopic examination revealed bone tissues covering the surface of the implant and bridging the gap. New bone was formed in the pores also. Tricalcium phosphate implants showed new bone formation due to osteoconductive properties. They are biodegradable. It is suggested that tricalcium phosphate implants are viable treatment alternatives in management of large osteoperiosteal defects with minimal to no adverse effects.

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 381-387

 

 

Sub-acute toxicity of cultured mycelia of Himalayan entomogenous fungus Cordyceps sinensis (Berk.) Sacc. in rats

Harsahay Meena*, Kshetra Pal Singh, Prem Sing Negi & Zakwan Ahmed

Defence Institute of Bio-Energy Research, DRDO, Field Station, Pithoragarh 262 501, India

Received 25 July 2012; revised 15 February 2013

Oral administration of laboratory cultured mycelia powder of C. sinensis did not show any sign of toxicity as no significant change was observed in organ weight and serological parameters in rats. However, there was a significant increase in food intake, body weight gain and hematological parameters like WBC, RBC, Hb and lymphocytes in treated groups. Histopathology of vital organs also supported the non toxic effect of C. sinensis. The results conclude that laboratory cultured mycelia powder of C. sinensis is safe and non toxic up to 2g/ kg body weight dose.

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 388-392

 

 

Effect of UV-C on thylakoid arrangement, pigment content and nitrogenase activity in the cyanobacterium Microchaete sp.

J K Sahua,* & M imek a,b

aFaculty of Science, University of South Bohemia, Česk Budějovice, Czech Republic

bBiology Centre of the ASCR, v. v. i. Institute of Soil Biology, and Department of Ecosystem Biology,
Faculty of Science, University of South Bohemia, Česk Budějovice, Czech Republic

Received 6 June 2012; revised 15 February 2013

The effect of UV-C radiation on thylakoid arrangement, chlorophyll-a and carotenoid content and nitrogenase activity of the cyanobacterium Microchaete sp. was studied. Chlorophyll-a and carotenoid content increased gradually up to 48 h of UV-C exposure but declined with longer exposures. Nitrogenase activity decreased moderately with 6 to 12 h exposure and decreased substantially afterwards. When cells exposed to UV-C for 12 to 24 h, grown under fluorescent light for 144 h, nitrogenase activity increased to levels greater than in the control cells. The exposure of UV-C treated cells to fluorescent light, however, did not result in recovery of pigment content. In Microchaete sp. cells treated with UV-C for 144 h, thylakoid membranes became dense, were aggregated into bundles, and were surrounded by spaces devoid of cytoplasm.

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 393-399

 

 

Microalgae mediated synthesis of silver nanoparticles and their antibacterial activity against pathogenic bacteria

S S Sudha, Karthic Rajamanickam & J Rengaramanujam

Postgraduate and Research Department of Microbiology,

Dr. N.G.P. Arts and Science College,
An Institution of Kovai Medical Center and Hospital [KMCH], Coimbatore 641 048, India

Received 9 October 2012; revised 20 February 2013

Silver nanoparticles is known to have antimicrobial affects. Cyanobacteria isolates from muthupet mangrove includes Aphanothece sp, Oscillatoria sp, Microcoleus sp, Aphanocapsa sp, Phormidium sp, Lyngbya sp, Gleocapsa sp, Synechococcus sp, Spirulina sp with were set in compliance with their cellular mechanism of nano silver creation, and were investigated by UV-VIS spectrophotometer, Energy-dispersive X-ray (EDX) and scanning electron microscopy (SEM). Silver nanoparticles were spherical shaped well distributed without aggregation in solution with an average size of about 40- 80 nm. Synthesised nano silver had antibacterial production on various organisms that provoked various diseases in humans. The cellular metabolites of Microcoleus sp. only created nano silver and it enhanced the antibacterial activity against test pathogenic bacteria from MTCC (Proteus vulgaris, Salmonella typhi, Vibrio cholera, Streptococcus sp., Bacillus subtilis, Staphylococcus aureus, Escherichia coli.) The antimicrobial assay was performed using 0.001 M concentration of nano silver in well diffusion method with positive control of appropriate standard antibiotic discs Cephotaxime, Ampicillin, Tetracyclin, Cephalexin etc. Synthesised silver nanoparticles acted as an effective antimicrobial agent and proved as an alternative for the development of new antimicrobial agents to combat the problem of resistance.

 

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 400-405

 

 

A multilayered supramolecular self-assembled structure from soybean oil by
in situ polymerization and its applications

 

Varadharajan Kavitha & Arumugam Gnanamani*

Microbiology Division, CSIR-Central Leather Research Institute (CLRI), Adyar, Chennai 600 020, India

Received 10 September 2012; revised 3 January 2013

The present study emphasizes in situ transformation of soybean oil to self-assembled supramolecular multilayered biopolymer material. The said polymer material was characterized and the entrapment efficacy of both hydrophilic and hydrophobic moieties was studied. In brief, soybean oil at varying concentration was mixed with mineral medium and incubated under agitation (200 rpm) at 37 C for 240 h. Physical observations were made till 240 h and the transformed biopolymer was separated and subjected to physical, chemical and functional characterization. The maximum size of the polymer material was measured as 2 cm in diameter and the cross sectional view displayed the multilayered onion rings like structures. SEM analysis illustrated the presence of multilayered honeycomb channeled structures. Thermal analysis demonstrated the thermal stability (200 C) and high heat enthalpy (1999 J/g). Further, this multilayered assembly was able to entrap both hydrophilic and hydrophobic components simultaneously, suggesting the potential industrial application of this material.

 

 

 

Indian Journal of Experimental Biology

Vol. 51, May 2013, pp. 406

 

 

Letter to Editor

Protective effects of subchronic caffeine administration on cisplatin induced urogenital toxicity in male mice

Hamid Nasri