Indian Journal of Experimental Biology

ISSN : 0019-5189   CODEN : IJEB (A6) 41(2) 99-184 (2003)
VOLUME 41

NUMBER 2

FEBRUARY 2003

 

 

 

CONTENTS

Letter to the Editor

 

Role of vessel factor in oxygen uptake study (an addendum)

103

M S Misra

 

 

 

Hypothesis

 

Post-irradiation free radical generation : Evidence from the conversion of xanthine

dehydrogenase into xanthine oxidase

R K Kale

 

105

 

 

Papers

 

Virus induced chromosomal abnormalities in Chinese hamster lung cell line and human peripheral blood leukocyte culture

Smita P Paranjape & U V Wagh

 

112

 

 

Effect of dietary zinc deficiency on metallothionein concentration of epididymal

luminal fluids of weanling Wistar albino rats

Ranjana Agrawal & R S Bedwal

 

118

 

 

Generation and characterization of monoclonal antibodies to protective antigen of

Bacillus anthracis

K S R Sastry, U Tuteja & H V Batra

 

123

 

 

Growth-associated production and characterization of poly (3-hydroxybutyric acid) of Azotobacter beijerinckii DAR-102

A Manna & A K Paul

 

129

 

 

Enzymatic and non-enzymatic antioxidants in selected Piper species

J Karthikeyan & P Rani

135

 

 

Simple and economical assay systems for evaluation of phosphinothricin resistant transgenics of sorghum, Sorghum bicolor. (L.) Moench., and pearl millet, Pennisetum glaucum (L.) R. Br.

D Harshavardhan, B Santha, T S Rani, K Ulaganathan, T Y Madhulety, C Laxminarayana & N Seetharama

 

 

141

 

 

Genetic transformation of Robinia pseudoacacia by Agrobacterium tumefaciens

Kamlesh Kanwar, Anju Bhardwaj, Sandhya Agarwal & D R Sharma

149

 

 

In vitro propagation of three commercial cut flower cultivars of Anthurium andraeanum Hort.

Dominic Joseph, K P Martin, Joseph Madassery & V J Philip

 

154

 

 

Effect of NaCl on leaf salt secretion and antioxidative enzyme level in roots of a

mangrove, Aegiceras corniculatum

Sujatarani Mishra & A B Das

 

160

 

 

Notes

 

Macromolecular synthesis in wing discs of Spodoptera mauritia Boisd : Effects of a juvenile hormone analogue

K Safarulla, Benny T M & V S K Nair

 

167

 

 

Response related enzymatic changes in ovaries of superovulated mice

Ranjna S Cheema, C K Dhanju & J S Matharoo

171

 

 

Effect of 24-epibrassinolide on pollen viability during heat-stress in tomato

Ishwar Singh & Mariko Shono

174

 

 

Detection of spores of Bacillus anthracis from environment using polymerase chain reaction

Syed Imteyaz Alam, Gauri Shankar Agarwal, Dev Vrat Kamboj, Ganga Prasad Rai & Lokendra Singh

 

177

 

 

Effect of methanolic leaf extract of Cissampelos mucronata A. Rich against

indomethacin induced ulcer in rats

S V Nwafor & P A Akah

 

181

 

 

Obituary

 

Professor A N Chakrabarty (1930-2002)

184

   
Author Index
 

 

 

 

Announcement

 

 

Indian Pharmacology Society

PP Surya Kumari Prize

 

PP Surya Kumari prize is awarded every year by the Indian Pharmacology Society (IPS) for the best paper on ‘Diabetes mellitus and other endocrinal and metabolic disorders’ published in any journal during last five years. The prize is open to Indian scientists working in Indian laboratories. The award is presented to the winner at the annual conference of IPS.

 

Those desirous of competing for the prize for the year 2002 may submit five reprints/copies of the paper, published during 1998-2002 to Professor R Raveendran, Convenor, PP Surya Kumari Prize, and The Chief Editor, Indian Journal of Pharmacology, Department of Pharmacology, Jawaharlal Institute of Postgraduate Medical Education and Research (JIPMER), Pondicherry 605 006, India, before

31 March 2003.

 

  Phone : 0413-22711969; Fax : 0413-2272067; E-mail : ijp@jipmer.edu.

 

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 103-104

   

 

Letter to the Editor

 

Role of vessel factor in oxygen uptake study (an addendum)

M S Misra

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 105-111

   

 

Hypothesis

 

 

Post-irradiation free radical generation: Evidence from the conversion of xanthine dehydrogenase into xanthine oxidase

R K Kale

 

The xanthine oxidoreductase (XOR) system which consists of xanthine dehydrogenase (XDH) and xathine oxidase (XO), is one of the major sources of free radicals in biological systems. The XOR system is pre-dominantly present as XDH in normal tissues and converts into the free radical generating XO-form in the damaged tissue. Therefore, the XO-form of the XOR system is expected to be mainly found in radiolytically damaged tissues. In such an event, XO may catalyze the generation of free radicals and potentiate radiation effects in the post-irradiation period. Recent findings on the effect of ionizing radiation on the XOR system in the liver of mice, peroxidative damage and lactate dehydrogenase support this possibility. From these results it has been hypothesized that free radical generating systems could be activated in the radiolytically damaged cell and in turn contribute to the cause and complications of late effects and their persistence in post-irradiation period. This aspect may have great significance in the understanding of radiation – induced damages. It may also have serious implication in various fields like radiation therapy, health physics, carcinogenesis, space travelling radiation exposures and post nuclear accident care. Further, it is suggested that efforts need to be made to search more system(s) which could be activated particularly at lower doses of radiation to generate free radicals in the post-exposure period.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 112-117

 

 

Papers

 

 

Virus induced chromosomal abnormalities in Chinese hamster lung cell line and human peripheral blood leukocyte culture

Smita P Paranjape

and

U V Wagh

 

Received 11 June 2002; revised 17 December 2002

 

 

Chinese hamster lung (CHL) cells were susceptible to Herpes Simplex type-1 and Chandipura viruses; which induced chromosomal abnormalities in these cells. Chromosomal changes induced in these cells were specific. The cells were refractory to measles virus and chromosomal abnormalities were not detected after inoculation of the virus. On the other hand human peripheral blood (HPB) leukocytes were susceptible to all the 3 viruses studied and exhibited chromosomal abnormalities upon infection. The aberrations induced in HPBL cultures were random. The results suggest that a virus could induce chromosomal changes only in susceptible cells. This is the first report of comparative in vitro study on chromosomes.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 118-122

 

 

Effect of dietary zinc deficiency on metallothionein concentration of epididymal luminal fluids of weanling Wistar albino rats

Ranjana Agrawal & R S Bedwal

 

Received 6 February 2002; revised 4 December 2002

 

 

Metallothionein (MT) and zinc concentrations have been estimated in luminal fluids of caput/corpus and cauda epididymis and serum of zinc deficient (ZD), pairfed (PF) and control — ad libitum fed (ZC) groups of Wistar rats. MT decreased significantly in luminal fluids of caput corpus and cauda epididymis and serum of zinc deficient rats as compared to their respective controls. However, the decrease was non-significant in luminal fluids of corpus epididymis and serum of 4-weeks zinc deficient animals as compared to their control. Zinc levels also declined significantly in luminal fluids of epididymis and serum of zinc deficient rats as compared to their respective pairfed and control groups. Thus zinc deficiency state reduces zinc and MT concentrations in luminal fluid of epididymis and serum.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 123-128

 

 

Generation and characterization of monoclonal antibodies to protective antigen of Bacillus anthracis

K S R Sastry, U Tuteja & H V Batra

 

Received 20 June 2001: revised 20 November 2002

 

 

Monoclonal antibodies (MoAbs) were generated following immunization of BALB/c mice with protective antigen (PA) of B. anthracis. Five clones reactive to this protein were stabilized and preserved. These MoAbs could detect nanogram levels of PA when tested in ELISA. In Western blotting, they reacted with all PA preparations tested and no cross-reactivity was observed with lethal factor, edema factor of B.anthracis and with other organisms. These MoAbs could detect PA from 22 confirmed clinical isolates of B. anthracis on Western blotting and hold promise for direct detection of PA in clinical samples for diagnosing anthrax.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 129-134

 

 

Growth-associated production and characterization of poly (3-hydroxybutyric acid) of Azotobacter beijerinckii DAR-102

A Manna & A K Paul

 

Received 20 March 2002; revised 20 August 2002

 

 

Production of poly (3-hydroxybutyric acid) [P(3HB)] by Azotobacter beijerinckii DAR-102 isolated in this laboratory has been optimized under batch-culture. The accumulatad polymer attained 58% of cell dry mass during mid-stationary phase with an yield of 0.58 g/l when grown in nitrogen-free medium. The optimum concentration of glucose and fructose for P(3HB) production was 3% (w/v) and 2% (w/v) respectively while that of casamino acid and tryptose was 0.1% (w/v). Phosphate at a concentration suboptimal for growth and limitation of oxygen in the medium favoured P(3HB) accumulation. The production of P(3HB) was maximum with an inoculum dose of 4% (v/v). The accumulated polymer was isolated by direct chloroform extraction of the dry cell mass and purified by precipitation with diethyl ether. The purified polymer has been characterized in terms of its solubility properties, melting temperature, and UV-, IR- and NMR-spectroscopic analyses.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 135-140

 

 

Enzymatic and non-enzymatic antioxidants in
selected Piper species

J Karthikeyan

and

P Rani

 

Received 30 April 2002; revised 9 December 2002

 

 

Piper species, commonly used in diet and traditional medicine were assessed for their antioxidant potential. Catalase activity was predominated in Piper longum, followed by Piper cubeba, green pepper, Piper brachystachyum and Piper nigrum. P. nigrum was richest in glutathione peroxidase and glucose-6-phosphate dehydrogenase, green pepper was richest in peroxidase and vitamin C while vitamin E was more in P. longum and P. nigrum. P. brachystachyum and P. longum were rich sources of vitamin A. All the Piper species had GSH content of around 1 to 2 nM/g tissue. The antioxidant components of Piper species constitute a very efficient system in scavenging a wide variety of reactive oxygen species. Antioxidant potential of Piper species was further confirmed by their ability to curtail in vitro lipid peroxidation by around 30-50% with concomitant increase in GSH content.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 141-148

 

 

Simple and economical assay systems for evaluation of phosphinothricin resistant transgenics of sorghum,
Sorghum bicolor. (L.) Moench., and pearl millet,
Pennisetum glaucum (L.) R. Br.

D Harshavardhan, B Santha, T S Rani, K Ulaganathan, T Y Madhulety, C Laxminarayana & N Seetharama

 

Received 15 April 2002;revised 18 November 2002

 

 

Five simple and rapid methods for evaluation of sorghum and pearl millet transgenics resistant to herbicide phosphinothricin (used as selectable marker) were studied. For rapid in vitro selection, three assays (establishment of sensitivity curves for embryogenic calli, determination of lethal doses for seed germination, and a rapid screening of cut young leaves based on the colour change of the medium) were established. For rapid screening of transgenic progeny, effects of in vivo Basta leaf spray and dip tests were studied at three different morphological stages. For all the above assays, LD50 and LD100 values were higher for pearl millet than sorghum. However, in both the crops, genotype effect was not significant. The assays standardized in the study were found to be effective for rapid, economical and mass-scale identification and characterization of transgenic plants of sorghum and pearl millet.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 149-153

 

 

Genetic transformation of Robinia pseudoacacia by
Agrobacterium tumefaciens

Kamlesh Kanwar, Anju Bhardwaj, Sandhya Agarwal & D R Sharma

 

Received 26 March 2002; revised 2 December 2002

 

 

Transgenic Robinia pseudoacacia plants were obtained by Agrobacterium tumefaciens mediated gene transfer.  Agrobacterium strain LBA4404 harbouring a binary vector that contained the chimeric neomycin phosphotransferase II (NPTII) and b-glucuronidase (GUS) genes was co-cultivated with hypocotyl segments of in vitro raised seedlings of Robinia.  Parameters important for high efficiency regeneration and transformation rates included type of explant, pre-conditioning of explants and appropriate length of co-cultivation period with Agrobacterium. A transformation frequency 16.67% was obtained by 48 hr of pre-conditioning followed by 48 hr of co-cultivation.  Transformed tissue was selected by the ability to grow on kanamycin containing medium. Successful regeneration was followed after histochemical GUS assay for the detection of transgenic tissue.  This transformation procedure has the potential to expand the range of genetic variation in Robinia.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 154-159

 

 

In vitro propagation of three commercial cut flower cultivars of
Anthurium andraeanum Hort.

Dominic Joseph, K P Martin, Joseph Madassery & V J Philip

 

Received 9 September 2002; revised 26 November 2002

 

 

In vitro propagation of Anthurium andraeanum Hort. cut flower cultivars viz. Lima White, Tropical White and Tropical Red through organogenesis using mature plant derived leaf explants was established on Murashige and Skoog (MS) medium fortified with different growth regulators. Cultivar, stage and different regions of the source leaf, and type of growth regulators significantly influenced callus induction. Explants from folded brown leaves were superior in induction of callus. Half strength MS medium fortified with 0.88 µM of benzyladenine (BA), 0.9 µM of 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.46 µM  of kinetin (Kn) at pH 5.5 was most effective for callus induction. Transfer of callus to medium with 0.54 mM of NAA in place of 2,4-D induced higher number of shoots. Subsequent cultures displayed enhanced rate of shoot initiation and multiplication. Transfer of shoots onto half strength MS medium supplemented with 0.54 mM of NAA favoured rooting of shoots. Cultivar Tropical White was superior in callus, shoot and root induction compared to Lima White and Tropical Red. Plantlets after acclimation in greenhouse were transferred to net-house, that exhibited ninety seven per cent survival. Plants flowered normally between 12 and 15 months and were morphologically similar to that of the mother plants.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 160-166

 

 

Effect of NaCl on leaf salt secretion and antioxidative enzyme level in roots of a mangrove, Aegiceras corniculatum

Sujatarani Mishra & A B Das

 

Received 4 June 2002; revised 20 November 2002

 

 

Short-term salt (NaCl) treatment on Aegiceras corniculatum in roots and leaves showed no change in fresh and dry weight of leaves, roots and leaf area. There was no significant change in total soluble root protein, photosynthetic pigments of leaves and spectral characteristics of thylakoids. However, the activity of antioxidative enzymes (catalase, ascorbate peroxidase and guaiacol peroxidase) in roots decreased by 72, 58 and 80% respectively after 96 hr of treatment (300 mM of NaCl). Secretion of salts from the leaf salt glands and salt accumulation on upper surface of the leaves were quantified that revealed linear increase of salt secretion of leaf with increase in period of salt treatment. It was concluded that loss of activities of antioxidative enzymes at high salt treatment, caused leaf senescence in spite of high rates of salt secretion by Aegiceras corniculatum.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 167-170

 

Notes

Macromolecular synthesis in wing discs of Spodoptera mauritia Boisd : Effects of a juvenile hormone analogue

K Safarulla, Benny T M & V S K Nair

 

Received 11 July 2002; revised 20 November 2002

 

 

Last instar larvae of S. mauritia treated topically on day 0, day 1, day 2 and day 3 with a daily dose of 25 mg juvenile hormone analogue (JHA) moulted into supernumerary larvae. The imaginal discs of the supernumerary larvae especially those of mouthparts and thoracic appendages showed pupal characteristics. However the wing discs, which showed only partial differentiation, were uneverted and highly tanned. In an effort to provide an explanation to this anomaly the RNA, DNA and protein profile in the wing discs of supernumerary larvae were studied. Quantitative analysis of DNA, RNA and protein showed a considerable increase in the amount of DNA and protein and a decline in RNA level. SDS-PAGE analysis of wing disc proteins of JHA treated larvae showed a reduction in the expression of many major proteins that were predominant in the wing discs of control larvae. The results suggest that JHA induced inactivation of genes involved in the synthesis of proteins needed for evagination process may be responsible for the formation of uneverted, partially differentiated pupal wing discs in supernumerary larvae.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 171-173

 

 

Response related enzymatic changes in ovaries of
superovulated mice

Ranjna S Cheema, C K Dhanju & J S Matharoo

 

Received 1 February 2002; revised 25 November 2002

 

 

Adult female mice were superovulated with PMSG followed by HCG and 140 blastocysts and 69 morulae were recovered from 24 mice. On the basis of the response, mice were divided into six groups; non responders, 1-5, 6-10, 11-20, 21-30 and >30 embryos. The ovaries of the animals were pooled group wise, homogenized in PBS (pH 7.4) and after centrifugation for 10-15 minutes, the supernatant was analyzed for the enzymes, guanine oxaloacetate transaminase (GOT), guanine pyruvate transaminase (GPT), acid phosphatases (ACP) and alkaline phosphatases (AKP). Acid and alkaline phosphatase activities did not show any variation in relation to response to superovulation but GOT and GPT showed significantly increased activity in response to induction of superovulation. A statistically significant positive correlation was found between GOT and GPT activities and the superovulatory response in mice.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 174-176

 

 

Effect of 24-epibrassinolide on pollen viability during
heat-stress in tomato

Ishwar Singh & Mariko Shono

 

Received 5 August 2002; revised 11 October 2002

 

 

Tomato pollen when treated with 24-epibrassinolide (EBR), it was observed that in vitro pollen germination was more tolerant to high temperature. A significantly higher in vitro pollen germination, enhanced tube growth and low pollen bursting were observed in presence of EBR at 35°C. Since a preconditioning treatment of tomato plant was not required for these effects, it is concluded that EBR treatment increased basic thermotolerance of germinating pollen. This study provided the first evidence for EBR induced thermotolerance in germinating pollen.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 177-180

 

 

Detection of spores of Bacillus anthracis from environment using
polymerase chain reaction

Syed Imteyaz Alam, Gauri Shankar Agarwal, Dev Vrat Kamboj, Ganga Prasad Rai & Lokendra Singh

 

Received 7 June 2002; revised 22 November 2002

 

 

A sensitive PCR based detection of Bacillus anthracis spores from environment was standardized. Specific 1247bp amplicon could be detected with template concentration as low as 13 pg. Sensitivity was enhanced to 10 fold by nesting with second set of primers, forming 208bp amplicon. Extraction of DNA from spores purified from soil samples by aqueous polymer two-phase system followed by partial germination and freeze-thaw treatment yielded best results. Soil sample spiked with spores (8´102/g of sample) could be detected with this method.

 

 

 

Indian Journal of Experimental Biology

Vol. 41, February 2003, pp. 181-183

 

 

Effect of methanolic leaf extract of Cissampelos mucronata A. Rich
against indomethacin induced ulcer in rats

S V Nwafor & P A Akah

 

Received 7 September 2001; revised 9 December 2002

 

 

Five fractions (F1 – F5) isolated from the methanolic leaf extract of Cissampelos mucronata A. Rich were investigated for antiulcer activity. At the dose of 450 mg/kg, they showed varying degree of protection against ulcer induced by indomethacin; the order of protection being F1>F4>F5>F2>F3. The antiulcer potency of F1 and F2 is comparable with that of cimetidine (100 mg/kg, ip). Inhibition of gastric mucosal damage may partly contribute to the antiulcer activity of the fractions.

 

 

 

AUTHOR INDEX

 

 

Agarwal Gauri Shankar

177

Matharoo J S

171

Agarwal Sandhya

149

Misra M S

103

Agrawal Ranjana

118

Mishra Sujatarani

160

Akah P A

181

 

 

Alam Syed Imteyaz

177

Nair V S K

167

 

 

Nwafor S V

181

Batra H V

123

 

 

Bedwal R S

118

Paranjape Smita P

112

Benny T M

167

Paul A K

129

Bhardwaj Anju

149

Philip V J

154

 

 

 

 

Cheema Ranjna S

171

Rai Ganga Prasad

177

 

 

Rani P

135

Das A B

160

Rani T S

141

Dhanju C K

171

 

 

Dominic Joseph

154

Safarulla K

167

 

 

Santha B

141

Harshavardhan D

141

Sastry K S R

123

 

 

Seetharama N

141

Kale R K

105

Sharma D R

149

Kamboj Dev Vrat

177

Shono Mariko

174

Kanwar Kamlesh

149

Singh Ishwar

174

Karthikeyan J

135

Singh Lokendra

177

 

 

 

 

Laxminarayana C

141

Tuteja U

123

 

 

 

 

Madassery Joseph

154

Ulaganathan K

141

Madhulety T Y

141

 

 

Manna A

129

Wagh U V

112

Martin K P

154