Indian Journal of Experimental Biology


Total visitors:521 since 18-8-05

ISSN : 0019-5189

CODEN : IJEB (A6) 43(9)  755-840 (2005)

VOLUME 43

NUMBER 9

SEPTEMBER 2005

 

CONTENTS

 

Papers

 

      Identification of 38kDa Brugia malayi microfilarial protease as a vaccine candidate for lymphatic filariasis
K N Krithika, Pankaj Dabir, Sandeep Kulkarni, V Anandharaman & M V R Reddy

 


759

      Protective role of allicin and L-ascorbic acid against the genotoxic damage induced by chlormadinone acetate in cultured human lymphocytes
Yasir Hasan Siddique & Mohammad Afzal

 


769

      Effect of spirulina and Liv-52 on cadmium induced toxicity in albino rats
K Jeyaprakash & P Chinnaswamy

 

773

      Biochemical and histological studies on H2-receptor antagonist ranitidine-induced
hepatotoxicity in rats
Faried A E Hemieda, El-Sayed K Abdel-Hady & Mostafa A Abou Elnga

 


782

 

      Short-term zinc deficiency in diet induces increased oxidative stress in testes and
epididymis of rats
N Nair, S Bedwal, S Prasad, M R Saini & R S Bedwal

 


786

      Effects of calcium channel blocker, mibefradil, and potassium channel opener, pinacidil, on the contractile response of mid-pregnant goat myometrium
Gurupada Mandi, S N Sarkar, S K Mishra & V Raviprakash

 


795

      Endophytic colonization and in planta nitrogen fixation by a diazotrophic Serratia sp.
in rice
G S Sandhiya, T C K Sugitha, D Balachandar & K Kumar

 


802

      A new approach for enhanced multiplication of arbuscular mycorrhizal fungi and isolation of ITS regions from Glomus deserticola and Laccaria fraterna
Padmanabhan Chellappan, Natarajan Mohan, Chellappa Ramesh, Thangaswamy Selvaraj, Muthaiyan Arunachalam & Ayyamperumal Mahadevan

 


808

      Polymorphism in Alnus based Frankia of Darjeeling
Manprit Bajwa, Amab Sen & Balwinder Singh Bajwa

813

      Genetic diversity and differentiation of Indian isolates of Phytophthora infestans as revealed by RAPD analysis
Ila Atheya, B P Singh, S K Chakrabarti & D Pattanayak

 


817

      Increase in NADH-glutamate dehydrogenase activity by mercury in excised bean
leaf segments
Priyanka Gupta & Rekha Gadre

 


824

      Rapid in vitro propagation of the threatened endemic orchid, Ipsea malabarica (Reichb.f.) J D Hook through protocorm-like bodies
K P Martin & Joseph Madassery

 

 

829

Notes

 

      Antibacterial activity of Ocimum sanctum L. fixed oil
Surender Singh, Manjusha Malhotra & D K Majumdar

835

      Cloning and sequencing of 28 kDa outer membrane protein gene of
Brucella melitensis Rev. 1
Pallab Chaudhuri, S Vinoth Kumar, Rajeev Prasad, S K Srivastava & M P Yadav

 


838

 

 

Announcement

 

TOXOCON-1

758

 

 

 

 

 

 

 

 

 

 

 

Announcement

 

TOXOCON-1

28 November 2005, Cochin, India

 

         With a view to bring professionals and students engaged in various branches of toxicology together, TOXOCON-1‑the inaugural conference of the Indian Society of Toxicology, will be held on 28 November 2005 at Amrita Institute of Medical Sciences, Cochin. Details of the conference can be obtained from Dr V V Pillay, Organizing Secretary, TOXOCON-1, Department of Analytical Toxicology, Amrita Institute of Medical Sciences, Cochin 682 026, India. Phone: 0484-2804852 (O), 2807055 (R), 9895282388 (Mobile); E-mail: toxicology@medical.amrita.edu

 

 

 

 

AUTHOR INDEX

 

 

Abdel-Hady El-Sayed K

782

Afzal Mohammad

769

Anandharaman V

759

Arunachalam Muthaiyan

808

Atheya Ila

817

Bajwa Balwinder Singh

813

Bajwa Manprit

813

Balachandar D

802

Bedwal R S

786

Bedwal S

786

Chakrabarti S K

817

Chaudhuri Pallab

838

Chellappan Padmanabhan

808

Chinnaswamy P

773

Dabir Pankaj

759

Elnga Mostafa A Abou

782

Gadre Rekha

824

Gupta Priyanka

824

Hemieda Faried A E

782

Jeyaprakash K

773

Krithika K N

759

Kulkarni Sandeep

759

Kumar K

802

Kumar S Vinoth

838

Madassery Joseph

829

Mahadevan Ayyamperumal

808

Majumdar D K

835

Malhotra Manjusha

835

Mandi Gurupada

795

Martin K P

829

Mishra S K

795

Mohan Natarajan

808

Nair N

786

Pattanayak D

817

Prasad Rajeev

838

Prasad S

786

Ramesh Chellappa

808

Raviprakash V

795

Reddy M V R

759

Saini M R

786

Sandhiya G S

802

Sarkar S N

795

Selvaraj Thangaswamy

808

Sen Arnab

813

Siddique Yasir Hasan

769

Singh B P

817

Singh Surender

835

Srivastava S K

838

Sugitha T C K

802

Yadav M P

838

 

 

 

KEYWORD INDEX

 

 

ACP

782

Actinorhizal plants

813

Allicin

769

ALT

782

Antibacterial

835

Antioxidant capacity

786

Antioxidants

769

AST

782

Bean leaf

824

Brucella melitensis

838

Bulbs

829

Cadmium chloride

773

Calcium channel blocker

795

Chlormadinone acetate

769

Chromosomal aberration

769

Cloning

838

Endophytes

802

Epididymis lipid peroxidation

786

Filariasis

759

Flavonoid

802

Frankia

813

Genetic diversity

817

Goat myomtrium

795

Glomus deserticola

808

Glutamate dehydrogenase

824

Glutamate synthase

824

GSH

786

GST

786

Hepatotoxicity

782

Hydroperoxides

786

Internal transcribed spacer (ITS)

808

Ipsea malabasica

829

Laccaria fraterna

808

l-Ascorbic acid

769

Linolenic acid

835

Liv-52

773

Malabar Daffodil orchid

829

Mercury effects

824

Mibefradil

795

Microfilarial soluble antigen

759

Mycorrhiza

808

Nitrogen fixation

802

NTSYpc

813

Ocimum sanctum fixed oil

835

Outer membrane protein

838

PCR-RFLP

813

Phaseolus vulgaris

824

Phytophthora infestans

817

Pinacidil

795

Potassium channel opener

795

Protease

759

Ranitidine

782

16 rRNA

813

RAPD analysis

817

Rat

782

Rhizome

829

Rice

802

S. aureus

835

Serratia

802

Serum

782

Sister chromatid exchanges

769

Spirulina

773

Testes

786

Thiobarbituric acid reactive substances (TBARS)

773

Vaccine

759

Zinc deficiency

786

 

 

 

 

 

 

 

 

Papers

 

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 759-768

 

 

Identification of 38kDa Brugia malayi microfilarial protease
as a vaccine candidate for lymphatic filariasis

K N Krithika, Pankaj Dabir, Sandeep Kulkarni, V Anandharaman & M V R Reddy

 

Received 7 January 2005; revised 30 May 2005

A FPLC purified 38kDa protease (Bm mf S-7) isolated from B. malayi microfilarial soluble antigen was identified. It showed pronounced reactivity with sera collected from 'putatively immune’ asymptomatic and amicrofilaraemic individuals residing in an endemic area for bancroftian filariasis. Further the immune protective activity of Bm mf S-7 antigen was evaluated in susceptible hosts, jirds (Meriones unguiculatus) against B. malayi filarial infection. The antigen showed 89% cytotoxicity against mf and 87-89% against infective (L3) larvae in in vitro antibody dependent cellular cytotoxicity Assay (ADCC) and in situ micropore chamber methods. Bm mf S-7 immunized jirds after challenge infection showed 81.5% reduction in the adult worm burden. The present study has shown that, the 38kDa microfilarial proteases (Bm mf S-7) could stimulate a strong protective immune response against microfilariae and infective larvae in jird model to block the transmission of filariasis. Analysis of IgG subclasses against Bm mf S-7 revealed a significant increase in IgG2 and IgG3 antibodies in endemic normals. Lymphocyte proliferation to Bm mf S-7 was significantly high in endemic normal group as compared to that in clinical and microfilarial carriers. Significantly enhanced levels of IFN-γ in the culture supernatant of PBMC of endemic normals followed by stimulation with Bm mf S-7 suggest that the cellular response in this group is skewed towards Th 1 type.

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 769-772

 

 

Protective role of allicin and L-ascorbic acid against the genotoxic damage induced by chlormadinone acetate in cultured human lymphocytes

Yasir Hasan Siddique & Mohammad Afzal

 

Received 10 December 2004; revised 16 March 2005

In our present study, different doses of allicin and L-ascorbic acid were tested against the genotoxic damage induced by chlormadinone acetate (CMA; 40 mM) using chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) as the parameters. Treatment with allicin and L-ascorbic acid resulted in reduction of CAs and SCEs. The results suggested a protective role of allicin and L-ascorbic acid against CMA induced genotoxic damage.

 

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 773-781

 

 

Effect of spirulina and Liv-52 on cadmium induced toxicity in albino rats

K Jeyaprakash & P Chinnaswamy

 

Received 7 July 2004; revised 26 May 2005

Oral administration of cadmium (6mg/kg body weight/day) as cadmium chloride (CdCl2) for 30 days resulted in a significant increase in thiobarbituric acid reactive substances (TBARS) level and a decrease in the levels of copper, zinc, iron, selenium, glutathione, superoxide dismutase, catalase, glutathione peroxidase when compared to normal control. Administration of either Liv-52 alone or in combination with spirulina produced a well pronounced protective effect in respect to these parameters in cadmium intoxicated rats. The protective effect of spirulina and Liv-52 in respect to biochemical changes were also confirmed by histopathological study in the liver and kidney sections.

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 782-785

 

 

Biochemical and histological studies on H2-receptor antagonist ranitidine-induced hepatotoxicity in rats

 

Faried A E Hemieda & El-Sayed K Abdel-Hady

 

and

Mostafa A Abou Elnga

 

Received 3 February 2005; revised 13 June 2005

This study was designed to investigate the hepatotoxicity of ranitidine treatment in dose levels of 10, 30, and 50 mg/kg b.wt. for 3 weeks period in male rats. The results showed some adverse changes in rats treated with either 10 or 30 mg/kg. Treatment with dose of 50 mg/kg produced marked increase in the activity of both acid phosphatase in liver and aspartate aminotransferase in serum and liver, with a tendency for increase in serum alanine aminotransferase activity. Also, a significant decrease in the serum activity of both amylase and alkaline phosphatase was noted. Microscopic examination of livers of the same animals revealed absence of some hepatic cells, pyknotic nuclei, dilatation of blood sinusoids, binucleated cells, and infiltration of lymphocytes. These biochemical and histological changes indicate that ranitidine when given chronically in high dose could produce hepatotoxicity in rats.

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 786-794

 

 

Short-term zinc deficiency in diet induces increased oxidative stress in testes and epididymis of rats

N Nair, S Bedwal, S Prasad, M R Saini & R S Bedwal

 

Received 6 January 2003; revised 26 April 2005

In order to determine the effects of Zinc deficient diet on oxidative stress in testis and epididymis, various parameters viz: total proteins, lipid peroxidation, hydroperoxides, antioxidant capacity and enzymatic activities are evaluated in rats fed on zinc deficient diet for 2, 4 and 6 weeks. Total proteins, water and lipid solouble antioxidant capacity decreased while lipid peroxidation (TBARS) and hydroperoxides concentration increased in testes, caput and cauda epididymis except in 2ZD (testes) where hydroperoxides revealed a significant decrease. GSH decreased in testes and caput and cauda epididymis. GPx and g-GT activities increased in testes and caput and cauda epididymis of zinc deficient rats. Further, GST increased in testes but exhibited decreases after 2 and 4 weeks and an increase after 6 weeks in caput and cauda epididymis of zinc deficient rats. GR activities decreased in testes but it increased in caput and cauda epididymis of zinc deficient rats. Thus, zinc deprivation results in increased sensitivity to oxidative stress. All these may have been as a consequence of increased ROS generation and/or decreased zinc dependent antioxidant processes.

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 795-801

 

 

Effects of calcium channel blocker, mibefradil, and potassium channel opener, pinacidil, on the contractile response of mid-pregnant goat myometrium

Gurupada Mandi, S N Sarkar, S K Mishra & V Raviprakash

 

Received 11 March 2005; revised 10 June 2005

The present study was undertaken to investigate the in vitro influence of mibefradil, a calcium channel blocker, and pinacidil, a potassium channel opener, on pregnant goat myometrial spontaneous rhythmic contractility and contractions induced with the agonist, oxytocin. Longitudinal strips from the distal region of uterus, collected from goats at mid-gestation, were mounted in an organ bath for recording isometric contractions. Mibefradil (10-8-10-4 M) or pinacidil
(10
-10-10-4 M), added cumulatively to the bath at an increment of 1 log unit, caused concentration-dependent inhibition of the spontaneous rhythmic contractions of isolated uterine strips. The rhythmic contraction was, respectively, abolished at 100 and 10 mM concentrations of mibefradil and pinacidil. In a concentration-dependent manner, mibefradil (1 and 10 mM) antagonized the contractions elicited with oxytocin (10-5-10-2 IU). Pretreatment of uterine strips with glibenclamide
(10
mM), a selective KATP channel blocker, caused a rightward shift of the concentration-response curve of pinacidil with a concomitant decrease in its pD2 value. Pinacidil (0.3, 1 and 3 mM), in a concentration-related manner, antagonized the oxytocin (10-5-10-2 IU)-induced contractile response. The inhibition of spontaneous rhythmic contractions and antagonism of oxytocin-induced contraction by mibefradil in the pregnant goat myometrium may be related to the antagonism of voltage-dependent Ca2+ channels, while by pinacidil suggests that KATP channel could be a therapeutic target for tocolysis.

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 802-807

 

 

Endophytic colonization and in planta nitrogen fixation by a
diazotrophic Serratia sp. in rice

G S Sandhiya, T C K Sugitha, D Balachandar & K Kumar

 

Received 15 February 2005; revised 31 May 2005

Nitrogen fixing endophytic Serratia sp. was isolated from rice and characterized. Re-colonization ability of Serratia sp. in the rice seedlings as endophyte was studied under laboratory condition. For detecting the re-colonization potential in the rice seedlings, Serratia sp. was marked with reporter genes (egfp and Kmr) using transposon mutagenesis. The conjugants were screened for re-colonization ability and presence of nif genes using PCR. Further, the influence of flavonoids and growth hormones on the endophytic colonization and in planta nitrogen fixation of Serratia was also investigated. The flavonoids, quercetin (3 mg/ml) and diadzein (2 mg/ml) significantly increased the re-colonization ability of the endophytic Serratia, whereas the growth hormones like IAA and NAA (5 mg/ml) reduced the endophytic colonization ability of Serratia sp. Similarly, the in planta nitrogen fixation by Serratia sp. in rice was significantly increased due to flavonoids. The inoculation of endophytic diazotrophs increased the plant biomass and biochemical constituents.

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 808-812

 

 

A new approach for enhanced multiplication of arbuscular mycorrhizal fungi and isolation of ITS regions from Glomus deserticola and Laccaria fraterna

Padmanabhan Chellappan, Natarajan Mohan, Chellappa Ramesh, Thangaswamy Selvaraj,, Muthaiyan Arunachalam and Ayyamperumal Mahadevan

 

Received 27 January 2005; revised 20 May 2005

Rootlets induced from the petiole base of L. purpureus, using IAA and kinetin was used for enhanced multiplication of arbuscular mycorrhizal (AM) fungus, G. deserticula. Using conserved short arbitrary oligonucleotides, as specific primers, we amplified the ITS-region, a molecular marker for fungal identification, from the genomic DNA extracted from cultured spores of G. deserticola, and genomic DNA extracted from the mycelium of L. fraterna. The capacity of fungal colonization and subsequent spore formation of G. deserticola, compared with the natural root system was evaluated. This technology would provide a simple way to multiply AM fungi and to produce spores without microbial contamination useful for further molecular characterization.

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 813-816

 

 

Polymorphism in Alnus based Frankia of Darjeeling

Manprit Bajwa, Arnab Sen & Balwinder Singh Bajwa

 

Received 16 November 2004; revised 3 May 2005

DNA samples extracted from the root nodules of Alnus nepalensis, collected from 10 different locations of Darjeeling hills, were used to assess the genetic diversity of Frankia. The DNA samples from the nodules of naturally growing plants were used as templates in PCR, targeting different genomic regions of Frankia, namely distal, middle and proximal parts of 16S rRNA gene and nifH-D IGS region with locus specific primers. The PCR products were digested with a number of frequent (4-base) cutter restriction endonucleases. Bands were scored as present (1) or absent (0) and the clustering was done using NTSYSpc. Distinct polymorphism was found among the nodules collected from different parts of the region and those of same geographic area. These results demonstrate that genetic diversity is indeed present among the naturally occurring Frankia of Darjeeling, India.

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 817-823

 

 

Genetic diversity and differentiation of Indian isolates of Phytophthora infestans as revealed by RAPD analysis

Ila Atheya, B P Singh, S K Chakrabarti and D Pattanayak

 

Received 31 January 2005; revised 20 April 2005

Sixty-seven isolates of Phytophthora infestans collected from Himalayan hill regions and subtropical planes of India were characterized by RAPD markers to assess diversity and differentiation based on location of origin. Ten random decamer primers generated 161 polymorphic fragments. Association of P. infestans isolates on the dendrogram and PCO plot revealed two clear grouping based on geographical location of origin-hill isolates and plane isolates. Quantification of diversity by Shannon index of diversity analysis demonstrated that most of the diversity was present with a particular population (hill or plane) of P. infestans isolates, with 85% variation being within and 15% being between hill and plane isolates. Subtropical plane isolates of P. infestans exhibited higher variability compared to hill isolates and they were more dispersed on the PCO plot. No clear differentiation of isolates based on mating type was reflected on the dendrogram and PCO plot.

 

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 824-828

 

 

Increase in NADH-glutamate dehydrogenase activity by mercury in excised
bean leaf segments

Priyanka Gupta & Rekha Gadre

 

Received 29 November 2004; revised 26 May 2005

Application of Hg to excised bean leaf segments increased the glutamate dehydrogenase (NADH – GDH) activity substantially. However, specific activity of the enzyme decreased at lower concentration of Hg, and increased to lesser extent at higher concentration of Hg. Mercury supply increased the glutamate synthase (NADH – GOGAT) activity also. Mercury supply increased the NADH – GDH activity in the presence of NH4NO3, but to a lesser extent than in the absence of NH4NO3. The specific activity of the enzyme decreased considerably at lower concentration of Hg, but increased significantly at higher concentration of Hg. An increase in NADH – GOGAT activity was observed in the presence of NH4NO3, but specific activity of the enzyme decreased marginally. Increase in GDH activity due to Hg remained unaffected by the supply of sucrose, but was reduced by glutamine and glutathione and enhanced by Al. The glutamate dehydrogenase (+Hg enzyme) from mercury treated leaf segments had higher value of S0.5 for NADH than the enzyme (-Hg enzyme) from material not treated with mercury indicating that Hg binding to enzyme prevented NADH binding to the enzyme possibly at thiol groups. However, + Hg enzyme has more reactivity, as apparent Vmax value was higher for it. It has been suggested that Hg activates the NADH-GDH enzyme in the bean leaf segments by binding to thiol groups of protein and pronounced increase in activity by Hg suggests a possible role of enzyme under Hg-stress.

 

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 829-834

 

 

Rapid in vitro propagation of the threatened endemic orchid, Ipsea malabarica (Reichb.f.) J D Hook through protocorm-like bodies

K P Martin & Joseph Madassery

Department of Biotechnology, University of Calicut, Calicut 673 635, India

Received 10 January 2005; revised 16 March 2005

Rapid propagation of I. malabarica (Reichb. f.) J D Hook, an endemic and endangered orchid of the Western Ghats of Kerala, India through conversion of axillary buds to protocorm-like bodies (PLBs), and subsequent plant regeneration was achieved. Growth regulators and sugar displayed significant influence in the induction of PLBs. In vitro derived shoots from field grown rhizomes of Ipsea cultured on Murashige and Skoog (MS) medium with 13.3 mM N6-benzyladenine (BA) containing 2% commercial grade sugar turned the axillary buds to PLBs within 25 days, and developed a mean of 33.1 PLBs within 50 days. Kinetin (KIN) did not induce PLBs, but facilitated axillary bud proliferation. Transfer of PLBs on medium having same concentration of BA and sugar facilitated rapid multiplication, and developed a mean of 47.5 PLBs. No decline of PLB proliferation was observed up to 10th subculture. Half strength MS medium with 6.97 mM KIN facilitated conversion of 98% PLBs to plantlets. On this media, a mean of 5.8 roots were also developed per shoot. Shoots developed bulbs during culture were grown to rhizomes. Increase of sugar to 6 or 8% hastened the development of bulbs/rhizomes. Reintroduction of PLB-derived plantlets in the natural habitat i.e. at Vellarimala (at 1300 m height) of the Western Ghats of Kerala was attempted as a means to assist in situ conservation. This is the first report of conversion of axillary buds to PLBs. The protocol enables to surmount the threat of extinction of this endemic and endangered orchid.

 

 

 

Notes

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 835-837

 

 

Antibacterial activity of Ocimum sanctum L. fixed oil

 

Surender Singh, Manjusha Malhotra & D K Majumdar

 

Received 29 November 2004; revised 27 May 2005

Ocimum sanctum fixed oil showed good antibacterial activity against Staphylococcus aureus, Bacillus pumilus and Pseudo­monas aeruginosa, where S. aureus was the most sensitive organism. Sesame and soyabean oils also showed moderate activity against S. aureus. Higher content of linolenic acid in
O. sanctum fixed oil could contribute towards its antibacterial activity. The antibacterial activity combined with anti-inflammatory and analgesic activities of the oil, could make it useful in inflammatory disorder resulting from staphylococcal infection.

 

 

 

Indian Journal of Experimental Biology

Vol. 43, September 2005, pp 838-840

 

 

Cloning and sequencing of 28 kDa outer membrane protein gene of Brucella melitensis Rev. 1

Pallab Chaudhuri , S Vinoth Kumar, Rajeev Prasad,
S K Srivastava & M P Yadav

 

Received 6 April, 2005

Brucella melitensis is an organism of paramount zoonotic importance. The 28 kDa outer membrane protein (OMP) is one of the immunodominant antigens of B. melitensis. The gene encoding 28 kDa OMP (omp28) has been amplified from
B. melitensis Rev. 1 strain. A PCR product of 753 bp, encoding complete omp28 gene of B. melitensis, was obtained. The gene was further cloned and sequenced. The nucleotide sequence of B. melitensis Rev. 1 strain showed substitution of 2 nucleotides from that of 16M strain.