Indian J Exp Biol

NOVEMBER 2002

CODEN: IJEB (A6)  40(11)  1209-1324  (2002)

ISSN: 0019-5189

 

 

 

Indian Journal of Experimental Biology

http : // www.niscair.res.in

 

VOLUME 40

NUMBER 11

NOVEMBER 2002

 

CODEN : IJEB (A6) 40(11) 1209-1324 (2002)

ISSN : 0019-5189

 

CONTENTS

Review Articles

 

 

Oxidants, antioxidants and carcinogenesis

Gibanananda Ray & Syed Akhtar Husain

1213

 

 

 

 

Oxidant-antioxidant system : Role and significance in human body

M Irshad & P S Chaudhuri

1233

 

Papers

 

 

Role of antioxidants in chronic fatigue syndrome in mice

Amanpreet Singh, Vivek Garg, Saraswati Gupta & Srinivas K Kulkarni

1240

 

 

 

 

Mentha piperita (Linn) leaf extract provides protection against radiation induced alterations in intestinal mucosa of Swiss albino mice

R M Samarth, M R Saini, J Maharwal, A Dhaka & Ashok Kumar

 

1245

 

 

 

 

Erythrocyte, lipid peroxidation and antioxidant enzymes in hypervitaminotic A rats and their modification by dietary protein

P K Karar, R Manavalan & G Rajagopal

 

1250

 

 

 

 

Effect of propriety herbal formulation against chronic carbon tetrachloride induced hepatotoxicity

Monika Bhadauria, Anjana Jadon, Abhilasha Sharma & Sangeeta Shukla

 

1254

 

 

 

 

Protection against photooxidative damage provided by enzymatic and non-enzymatic antioxidant system in sorghum seedlings

Sangeeta Sankhalkar & Prabhat Kumar Sharma

 

1260

 

 

 

 

Analgesia in phasic and tonic pain tests in a pharmacological model of autotomy

Suman Jain & Ratna Sharma

1269

 

 

 

 

Magnesium deficiency increases oxidative stress in rats

Chetan Parkash Hans, Dharam Paul Chaudhary & Devi Dayal Bansal

1275

 

 

 

 

Staphylococcus warneri BW 94 ¾ A new source of lipase

G S Walavalkar & M M Bapat

1280

 

 

 

 

 

Assessment of EMS-induced genotoxicity in the Indian climbing perch, Anabas
testudineus
: Cytogenetical vis-à-vis protein endpoints

B Guha & A R Khuda-Bukhsh

 

1285

 

 

Regeneration from mature and immature embryos and transient gene expression via Agrobacterium-mediated transformation in emmer wheat (Triticum dicoccum Schuble)

Jigyasa Khurana, Archana Chugh & Paramjit Khurana

 

 

1295

Notes

 

Storage excretion in the Indian apple snail, Pila globosa (Swainson), during aestivation

Madhura S Athawale & S Raghupathi Rami Reddy

1304

 

 

Impact of feeding ethanolic extracts of Achyranthes aspera Linn. on reproductive functions in male rats

K Sandhyakumary, R G Boby & M Indira

 

1307

 

 

Effect of whole body gamma radiation on hepatic LDH activity, lactate, pyruvate concentration and rate of oxygen consumption in Bufo melanostictus

J Mishra, B Mittra & A Mittra

 

1310

 

 

Anti-nociceptive effect of synthesized di-hydroxy flavones: Possible mechanism

K Girija, M Kannappa Reddy & S Viswanathan

1314

 

 

Book Reviews

 

Our posthuman nature: Consequences of the biotechnology revolution; Redesigning humans: Choosing our children’s genes

Suresh I S Rattan

 

Laboratory manual : Diagnostic methods for animal and poultry mycoplasmosis

H Rahman

NMDA antagonists as potential analgesic drugs (Progress in inflammation research); Anxiety disorders pathophysiology and pharmacological treatment

Salil K Bhattacharya

1317

 

 

 

1318

 

 

1318
1319

 

 

 

Conference Report

 

5th European Nitrogen Fixation Conference ¾ A report

G S Randhawa

1321

 

 

News Scan

 

Banana potassium and stroke; Calcium may cut colon cancer risk

M K Singhal

1322

 

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1213-1232

 

 

 

Review Article

 

Oxidants, antioxidants and carcinogenesis

Gibanananda Ray & Syed Akhtar Husain

 

Reactive oxygen metabolites (ROMs), such as superoxide anions (O2•-) hydrogen peroxide (H2O2), and hydroxl radical (•OH), malondialdehyde (MDA) and nitric oxide (NO) are directly or indirectly involved in multistage process of carcinogenesis. They are mainly involved in DNA damage leading sometimes to mutations in tumour suppressor genes. They also act as initiator and/or promotor in carcinogenesis. Some of them are mutagenic in mammalian systems. O2•-, H2O2 and •OH are reported to be involved in higher frequencies of sister chromatid exchanges (SCEs) and chromosome breaks and gaps (CBGs). MDA, a bi-product of lipid peroxidation (LPO), is said to be involved in DNA adduct formations, which are believed to be responsible for carcinogenesis. NO, on the other hand, plays a duel role in cancer. At high concentration it kills tumour cells, but at low concentration it promotes tumour growth and metastasis. It causes DNA single and double strand breaks. The metabolites of NO such as peroxynitrite (OONO-) is a potent mutagen that can induce transversion mutations. NO can stimulate O2-/H2O2/•OH-induced LPO. These deleterious actions of oxidants can be countered by antioxidant defence system in humans. There are first line defense antioxidants such as superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT). SOD converts O2•- to H2O2, which is further converted to H2O with the help of GPx and CAT. SOD inhibits •OH production. SOD also act as antipoliferative agent, anticarcinogens, and inhibitor at initiation and promotion/transformation stage in carcinogenesis. GPx is another antioxidative enzyme which catalyses to convert H2O2, to H2O. The most potent enzyme is CAT. GPx and CAT are important in the inactivation of many environmental mutagens. CAT is also found to reduce the SCE levels and chromosomal aberrations. Antioxidative vitamins such as vitamin A, E, and C have a number of biological activities such as immune stimulation, inhibition of nitrosamine formation and an alteration of metabolic activations of carcinogens. They can prevent genetic changes by inhibiting DNA damage induced by the ROMs. Therefore, these antioxidants may be helpful in the treatment of human cancer. However, detailed studies are required to draw a definite conclusion.

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1233-1239

 

 

Oxidant-antioxidant system : Role and significance in human body

M Irshad & P S Chaudhuri

 

Present article gives a holistic view of the causes, role and conrol of oxidative stress in the development and progression of various human diseases. Several types of reactive species are generated in the body as a result of metabolic reactions in the form of free radicals or non-radicals. These species may be either oxygen derived or nitrogen derived and called pro-oxidants. They attack macromolecules including protein, DNA and lipid etc. causing cellular / tissue damage. To counter their effect, the body is endowed with another category of compounds called antioxidants. These antioxidants are produced either endogenously or received from exogenous sources and include enzymes like superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase, minerals like Se, Mn, Cu and Zn, and vitamins like vitamin A, C and E. Other compounds with antioxidant activity include glutathione, flavonoids, bilirubin and uric acid etc.. In a healthy body, pro-oxidants and antioxidants maintain a ratio and a shift in this ratio towards prooxidants gives rise to oxidative stress. This oxidative stress may be either mild or severe depending on the extent of shift and remains the cause of several diseases such as cardiovascular diseases, neurological diseases, malignancies, renal diseases, diabetes, inflammatory problems, skin diseases, aging, respiratory diseases, liver diseases and different types of viral infections. As more and more reports are pouring-in, a lot of information is being unfolded about oxidative stress in relation to several other diseases.

 

Papers

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1240-1244

 

 

 

Role of antioxidants in chronic fatigue syndrome in mice

Amanpreet Singh, Vivek Garg, Saraswati Gupta & Srinivas K Kulkarni

 

Received 21 March 2002; revised 27 August 2002

The present study was carried out using mice model of chronic fatigue syndrome (CFS) in which mice were forced to swim everyday for 7 days for a 6 min session. There was a significant increase in despair behavior (immobility period) in saline treated mice on successive days. Treatment with potent antioxidants carvedilol (5 mg/kg, ip) and melatonin (10 mg /kg, ip) produced a significant reduction in immobility period. Similar results were observed with herbal products St. John’s Wort (Hypericum perforatum.L) (10 mg/kg, po) and GS-02 (20 mg /kg, po). Fluoxetine, a selective serotonin reuptake inhibitor produced a significant effect only on first and second day of its treatment. Biochemical analysis revealed that chronic swim test significantly increased lipid peroxidation and catalase levels in whole brains of mice. There was a decrease in the levels of super oxide dismutase (SOD) and glutathione reductase (GSH) in the brain. Administration of carvedilol, melatonin, GS-02 and St. John’s Wort restored the levels of lipid peroxidation and glutathione. The enzymes SOD and catalase were also restored. Fluoxetine affected the biochemical variables not to the same extent as other treatments. The findings of the present study suggest that oxidative stress might play a significant role in the pathophysiology of CFS. Thus antioxidants and herbal products like St. Johns wort and GS-02 could be useful in the treatment of CFS.

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1245-1249

 

 

Mentha piperita (Linn) leaf extract provides protection against radiation induced alterations in intestinal mucosa of Swiss albino mice

R M Samarth, M R Saini, J Maharwal, A Dhaka & Ashok Kumar

 

Received 23 April 2002; revised 13 August 2002

Intestinal protection in mice against radiation injury by M. piperita (1 g/kg body weight/day) was studied from day 1 to day 20 after whole body gamma irradiation (8 Gy). Villus height, goblet cells/villus section, total cells, mitotic cells and dead cells/crypt section in the jejunum are good parameters for the assessment of radiation damage. There was significant decrease in the villus height, number of total cells and mitotic cells/crypt section, whereas goblet cells and dead cells showed significant increase after irradiation. Mentha pretreatment resulted in a significant increase in villus height, total cells and mitotic cells, whereas goblet cells and dead cells showed a significant decrease from respective irradiated controls at each autopsy day. The results suggest that Mentha pretreatment provides protection against radiation induced alterations in intestinal mucosa of Swiss albino mice.

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1250-1253

 

Erythrocyte, lipid peroxidation and antioxidant enzymes in hypervitaminotic A rats and their modification by dietary protein

P K Karar & R Manavalan

 

and

G Rajagopal

 

Received 15 March 2002; revised 26 August 2002

Rats fed excess vitamin A showed decreased body weight gain and protein efficiency ratio. In rats fed low protein vitamin A level increased in liver but with an associated decrease in plasma. These changes were reversed in high protein fed state. The amount of protein in diet had little effect on haemoglobin level in erythrocyte, but excess vitamin A in diet significantly decreased haemoglobin level in erythrocyte. Lipid peroxidation (LP) increased in rats fed low protein and decreased in high protein fed rats. Rats fed high protein and excess vitamin A showed minimum level of LP. Result showed that high protein in diet increased the levels of antioxidant enzymes, catalase and superoxide dismutase (SOD) and that excess vitamin A supplementation functions synergistically with high protein in diet to increase antioxidant enzymes level.

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1254-1259

 

Effect of propriety herbal formulation against chronic carbon tetrachloride
induced hepatotoxicity

Monika Bhadauria, Anjana Jadon, Abhilasha Sharma & Sangeeta Shukla

 

Received 26 March 2002; revised 16 August 2002

Efficacy of propriety herbal formulation (PHF) against carbon tetrachloride (CCl4) induced liver damage was investigated in adult rats. Administration of CCl4 (0.2 ml/kg; ip) twice a week for 12 weeks resulted in significant elevation in serum transaminases activity. Level of reduced glutathione was significantly decreased. On the contrary, significant elevation was found in the hepatic lipid peroxidation level. Proliferation of fibroblast replaced the hepatic parenchyma cells in focal areas. Cell organelles like mitochondria, endoplasmic reticulum and nucleus showed severe degeneration after CCl4 exposure. PHF was effective in restoring the CCl4 induced biochemical and histological ultrastructural changes

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1260-1268

 

 

Protection against photooxidative damage provided by enzymatic and
non-enzymatic antioxidant system in sorghum seedlings

Sangeeta Sankhalkar & Prabhat Kumar Sharma

 

Received 30 January 2002; revised 30 July 2002

 

Effect of photoinhibition of sorghum leaves and isolated chloroplasts on chlorophyll fluorescence, peroxidation of thylakoid lipids and activity of antioxidant enzymes were studied. Photoinhibition of intact leaves and isolated chloroplasts decreased Fv/Fm ratio and qP, while qN increased. Photoinhibitory damage was more at 5oC than at 30o or 50oC. Peroxidation of thylakoid lipids was 5 times greater when photoinhibited at 50oC compared to control. Photoinhibition of chloroplasts under low oxygen condition or when supplemented with anti-oxidants (b-carotene, ascorbate and GSH) resulted in significantly less damage to photosynthesis (Fv/Fm ratio) and peroxidation level. Photoinhibition also resulted in many fold increase in the activity of superoxide dismutase (SOD) and ascorbate peroxidase (APX) and decrease in catalase. Data presented here suggest that photoinhibition resulted in production of oxygen radicals and photoinhibition of chloroplasts in the presence of low oxygen level or when supplemented with antioxidants decreased the damage to Fv/Fm ratio and peroxidation level to a great extent since former prevented the formation of oxygen radicals and later could scavenge the oxygen radicals thus the protection. Increase activity of SOD and APX may also be to metabolise the oxygen radicals produced during photoinhibition treatment, thereby, protecting the seedlings against photooxidative damage.

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1269-1274

 

Analgesia in phasic and tonic pain tests in a pharmacological model of autotomy

Suman Jain & Ratna Sharma

 

Received 22 May 2002; revised 21 August 2002

Self-mutilation or self-injurious behaviour is a well known behavioural disorder in humans. The proposition that this behaviour in animals is a response to chronic pain of peripheral nerve injury has been met with controversy. In the present study a pharmacological model, which produces no sensory or motor loss was used to study how autotomy is related to pain. In a group of rats autotomy was induced by amphetamine in phenoxybenzamine and reserpine treated animals. The pain tests, both phasic and tonic were then performed. The results of this study showed that a total analgesia was produced in both phasic and tonic pain tests, in animals that exhibited autotomy. Injection of naloxone in these animals prevented autotomy. A correlation between autotomy and no pain is suggested in this pharmacological model of autotomy.

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1275-1279

 

Magnesium deficiency increases oxidative stress in rats

Chetan Parkash Hans, Dharam Paul Chaudhary & Devi Dayal Bansal

 

Revised 4 April 2002; revised 27 August 2002

Magnesium deficiency has been implicated in the development of atherosclerosis and late diabetic complications, diseases often associated with increased oxidative stress. Present study was carried out to examine the effect of magnesium deficiency on oxidative stress and total radical trapping antioxidant parameter (calculated) in rats and correlate it with the development of free radical mediated diseases. Male Wistar rats were divided into two groups and pair fed for six weeks with low magnesium diet (70 mg/kg) and control diet (990 mg/kg) prepared synthetically. Deionized water was given ad libitum. Low magnesium diet caused a significant decrease in plasma and red blood cell magnesium levels. A marked increase in plasma malondialdehyde and corresponding decrease in total radical trapping antioxidant parameters (calculated) were observed in the low magnesium diet group than control group. The level of plasma glucose increased moderately in the low magnesium diet group. Hypertriglyceridemia and significantly decreased plasma HDL (high density lipoprotein)-cholesterol levels were observed in the low magnesium diet group. The results clearly demonstrate that magnesium deficiency is associated with increased oxidative stress through reduction in plasma antioxidants and increased lipid peroxidation suggesting that the increased oxidative stress may be due to increased susceptibility of body organs to free radical injury.

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1280-1284

 

Staphylococcus warneri BW 94 ¾ A new source of lipase

G S Walavalkar & M M Bapat

 

Received 9 April 2001; revised 5 August 2002

Staphylococcus isolated from a common Indian sweet viz. basundi was tested for its ability to produce lipase. The colorless zone of hydrolysis around the colony grown on Baird Parker agar containing egg yolk produced extracellular lipase. Colony morphology, coagulase production, haemolysis, acid production in carbohydrate medium and enzyme activity studies showed that the organism was Staphylococcus warneri. Growth of S. warneri was obtained after 11hr at 37oC, pH 7.5, while the maximum production of lipase was obtained at 30oC at pH 6.5 after 9hr of incubation. Agitation did not increase lipase production. A sudden fall in the activity of lipase was noted after 11hr. Addition of sucrose which is a growth stimulant for Staphylococcus, did not stimulate production of lipase by these organisms. Also, addition of oleic acid, Tween 80 or ethanol did not stimulate formation of lipase.

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1285-1294

 

Assessment of EMS-induced genotoxicity in the Indian climbing perch,
Anabas testudineus : Cytogenetical vis-à-vis protein endpoints

B. Guha & A R Khuda-Bukhsh

 

Received 18 March 2002; revised 20 June 2002

Genotoxic effects of EMS have been assessed in fish, A. testudineus, using widely accepted cytogenetic protocols like chromosome aberrations, nuclear anomalies in red blood cells and abnormal sperm head morphology. In addition, gel electrophoretic protein profiles and total protein contents in nine selected tissues were analysed for evaluating their utility as potential indicators of genotoxicity. EMS not only caused chromosomal aberrations in somatic cells, nuclear anomalies in red blood cells, and increased incidence of sperm with abnormal head morphology, but also altered significantly both protein profiles and total protein contents in all tissues tested vis-à-vis suitable controls, indicating relevance of protein data in genotoxicity assessment.

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1295-1303

 

Regeneration from mature and immature embryos and transient gene expression via Agrobacterium-mediated transformation in emmer wheat
(Triticum dicoccum Schuble)

Jigyasa Khurana, Archana Chugh & Paramjit Khurana

 

Received 14 June 2002; revised 5 August 2002

The present study establishes a regeneration protocol and optimizes conditions for Agrobacterium-mediated transformation of the tetraploid emmer wheat, Triticum dicoccum. Regeneration from mature and immature embryos was accomplished as a two-step process involving callus induction in the presence of 2,4-D followed by regeneration on a 2,4-D free, cytokinin-containing medium (RM1). Higher concentrations of 2,4-D (4 mg/l) though conducive for callusing (89.39% in mature embryos and 96% in immature embryos) proved detrimental for further regeneration. At lower 2,4-D (1 mg/ml) although callusing was suboptimal, (56.8% and 84% from mature and immature embryos, respectively) the regeneration response was the highest on RM1 medium (64.4% and 56.6% from mature and immature embryos, respectively). Overall, the regeneration response of immature embryos was lower than the mature embryos by 10-12%. Due to the ease of availability of mature embryos the mature embryo-derived calli were chosen as the target tissue for Agrobacterium-mediated transformation in the two Indian varieties DDK1001 and DDK1009. Histochemical GUS expression revealed the suitability of the mature embryo -derived calli for such investigations. Of the CaMV35S and Act1 promoters employed, the monocot promoter Act1 displayed higher GUS gene activity in the mature embryo derived calli when co-cultivated with LBA4404 (pBI101:: Act1).

Notes

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1304-1306

 

Storage excretion in the Indian apple snail, Pila globosa (Swainson),
during aestivation

Madhura S Athawale & S Raghupathi Rami Reddy

 

Received 1 March 2002; revised 31 July 2002

Uric acid accumulates in several tissues of the Indian apple snail, P. globosa, during aestivation. This accumulation is particularly high in the foot muscle and reproductive organs. Since the kidney, a tiny organ in the snail, has only a limited capacity to store uric acid, extrarenal tissues are used as storage depots of uric acid during aestivation. It is suggested that the aestivating snail, faced with a cleidoic situation, resorts to storage excretion, a phenomenon well documented in insects.


Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1307-1309

 

Impact of feeding ethanolic extracts of Achyranthes aspera Linn. on reproductive functions in male rats

K Sandhyakumary, R G Boby & M Indira

 

Received 8 August 2001; revised 20 August 2002

Feeding 50% ethanolic extract of A. aspera to male rats resulted in reduced sperm counts, weight of epididymis, serum level of testosterone and testicular activity of 3b-hydroxysteroid dehydrogenase, while motility of the sperm and activity of the HMG CoA reductase were not affected. Cholesterol level in the testis, incorporation of labelled acetate into cholesterol, 17-ketosteroids in urine and hepatic and fecal bile acids were increased. The results suggest that ethanolic extract of A. aspera caused reproductive toxicity in male rats and the action may be by suppressing the synthesis of androgen.

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1310-1313

Effect of whole body gamma radiation on hepatic LDH activity, lactate, pyruvate concentration and rate of oxygen consumption in Bufo melanostictus

J Mishra, B Mittra & A Mittra

 

Received 13 May 2002; revised 20 August 2002

Whole body Co60 gamma radiation induced changes in lactic dehydrogenase (LDH) activity, pyruvate, lactate content and rate of oxygen (O2) consumption in a tropical hibernating anuran (Bufo melanostictus). In 3.5 and 7 Gy treated groups, a significant increase in LDH activity and lactate/ pyruvate ratio was observed, whereas a significant decrease in O2 consumption rate was observed in treated animals on post-irradiation day (PID) 1, 5 and 10. Increase in LDH activity was observed on PID-1 in both the treated groups, reached to a peak on PID-5 in 7 Gy treated group and then declined on PID-10.

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1314-1316

 

 

Anti-nociceptive effect of synthesized di-hydroxy flavones: Possible mechanism

K Girija, M Kannappa Reddy & S Viswanathan

 

Received 4 March 2002; revised 30 May 2002

Renewed interest on the research on the flavonoids is gaining more importance. Earlier literature on flavonoids indicated a significant anti-nociceptive action for flavones and mono-substituted flavones. However, they exhibited a ceiling effect. The present study was undertaken by new synthesizing six disubstituted flavones (DHFs) since poly substituted ones are expected to produce more potent effect. Their anti-nociceptive effect and the role of opioid involvement were studied using acetic acid induced abdominal constriction assay. All the six DHFs administered in elicited a dose related inhibition of abdominal constrictions indicating the presence of the anti-nociceptive response. However, these substances also showed a similar ceiling effect. Like other flavonoid substances, they also utilized opioid pathways. It is suggested that these newly synthesized DHFs can be included along with other flavonoids while attempting clinical trial for analgesic use.

 

 

Book Reviews

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1317

 

 

Our posthuman nature: Consequences of the biotechnology revolution

Redesigning humans: Choosing our children’s genes

Suresh I S Rattan

 

 

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1318

 

 

Laboratory manual : Diagnostic methods for animal and poultry mycoplasmosis

H Rahman

 

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1319-1320

 

 

NMDA antagonists as potential analgesic drugs (Progress in inflammation research)

 

Anxiety disorders pathophysiology and pharmacological treatment

Salil K Bhattacharya

 

Conference Report

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1321

 

 

5th European Nitrogen Fixation Conference ¾ A Report

G S Randhawa

News Scan

Indian Journal of Experimental Biology

Vol. 40, November 2002, pp. 1322

 

 

Banana potassium and stroke

Calcium may cut colon cancer risk

M K Singhal