Indian J Exp Biol

SEPTEMBER 2002

CODEN: IJEB (A6) 40(9) 977-1092 (2002)

ISSN: 0019-5189

Indian Journal of Experimental Biology
http : // http : //
VOLUME 40
CODEN:IJEB (A6) 40(9) 977-1092 (2002) / NUMBER 9 /

SEPTEMBER 2002

ISSN: 0019-5189
CONTENTS
Review Article
Nitrogen control of bacterial signal production in Rhizobium meliloti-alfalfa symbiosis
Ilona Dusha / 981
Mini Review
DNA mismatch repair, microsatellite instability and cancer
Minal Vaish. & B Mittal / 989
Papers
Role of macrophage-colony stimulating factor and osteoclast differentiation factor in osteoclastogenesis of bone marrow derived stem cells
Mohamed Bayoumy, Uma Sankar & Natarajan Muthusamy / 995
Nuclear zinc in liver of cadmium and zinc treated rats
Ziad M Bataineh & Jamil R Al-Alami / 1001
Antidiabetic and antioxidant effects of S-methyl cysteine sulfoxide isolated from onions (Allium cepa Linn) as compared to standard drugs in alloxan diabetic rats
Kumud Kumari& KT Augusti / 1005
Effect of oxidative stress on serum and antioxidant enzymes in liver and kidney of rats and their modulation through dietary factors
H D Ramachandran, KNarasimhamurthy & P L Raina / 1010
Response of liver antioxidant system to taurine in rats fed high fructose diet
A T Anitha Nandhini, S D Balakrishnan & C V Anuradha / 1016
Efficiency of Coleus aromaticus extract in modifying cyclophosphamide and
mitomycin-C induced clastogenicity in mouse bone marrow cells
Shyama PrasadS, Prashantha Naik & K K Vijayalaxmi / 1020
Comparative immunopotentiating properties of saponin and incomplete Freund’s
adjuvant coupled to affinity purified larval antigen of Hyalomma anatolicum
anatolicum
S Ghosh, N K Singh & Preeti Rawat / 1026
-fructofuranosidase production by 2-deoxyglucose resistant mutants of Aspergillus niger in submerged and solid-state fermentation
BalasubramaniemAshokkumar & Paramasamy Gunasekaran / 1032
Infra-red spectroscopic analyses of banana waste degraded by oyster mushroom
G V Reddy, M P Shah, I L Kothari & A Ray† / 1038
A novel strategy of oxygen restriction by some diazotrophic enteric bacteria
V Kannan & P N Raju / 1043
In vitro plantlet regeneration from seedling nodal explants of Acacia catechu
Rohini Sahni& Shrish C Gupta / 1050
Esterase as molecular marker for salt tolerance in regenerated plants of rice, Oryza
sativa L
T S Swapna / 1056
Bioregulation of carbohydrate metabolism in relation to source-sink operation during grain-filling phase of growth in wheat
Parminder Sidhu & Rangil Singh / 1060
Notes
In vitro formation of organ specific proximate carcinogen of benzo(a)pyrene by rat homogenates
Sucheta Sharma & H M Dani / 1067
Impact of hypercholesterolemia on in vitro toxicity of N-nitrosodiethylamine: Effect on lipidperoxidation of blood and tissue
Gaurav Mittal, Manpreet Kaur & Giridhar Soni / 1071
Frequency of feeding and formation of bone growth marks in frog, Rana cyanophlyctis (Schn.)
S M Kumbar & K Pancharatna / 1074
Urinary proteins and pheromonal communication in mammals
S Achiraman & G Archunan / 1077
Ocimum sanctum aqueous leaf extract provides protection against mercury induced toxicity in Swiss albino mice
Mukesh Kumar Sharma, Madhu Kumar & Ashok Kumar / 1079
Production of glycolipids containing biosurfactant by Pseudomonas species
P Ellaiah,TPrabhakar, M Sreekanth, A Thaer Taleb, P Bhima Raju & V Saisha / 1083
Monoxenic in vitro production and colonization potential of AM fungus
Glomus intraradices
A Mohammad & A G Khan / 1087
Announcements
16th National Conference of Parasitology
International Conference on Chest Diseases and Allied Sciences
National Conference on Recent Trends in Plant Science Research / 1092

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 981-988

Review Article

Nitrogen control of bacterial signal production in
Rhizobium meliloti-alfalfa symbiosis

Ilona Dusha

Under nitrogen-depleted conditions nitrogen-fixing soil bacteria of the family Rhizobiaceae are able to induce symbiotic nodules on the roots of leguminous plants where bacteroids convert atmospheric nitrogen to ammonia. The presence of exogenous nitrogen source inhibits the development and the functioning of bacterium-plant symbiosis. Earlier experiments demonstrated that nitrate inhibited all stages of symbiotic interaction, affecting primarily the host functions. The investigation of the possible involvement of the microsymbiont in nitrogen regulation showed that two signalling steps were controlled by ammonium. The synthesis of the first bacterial signal, the Nod factor was repressed by ammonium. The nitrogen signal is conveyed to nodulation (nod) genes by the general nitrogen regulatory (ntr) system and by the nodD3-syrM self-amplifying system. The fine control also involves a negative regulatory factor, ntrR. When ntrR is mutated, more efficient nodule formation and nitrogen fixation is observed in symbiosis with alfalfa even in the presence of ammonium. The biosynthesis of the second bacterial signal succinoglycan is also controlled by ammonium. SyrM, a common regulatory factor for nod and exo gene expression, may contribute to the adjustment of the amount of succinoglycan and the ratio of its biologically active form.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 989-994

Mini Review

DNA mismatch repair, microsatellite instability and cancer

Minal Vaish. & B Mittal*

Department of Medical Genetics, Sanjay Gandhi Post Graduate Institute of Medical Sciences,
Raebareli Road, Lucknow 226 014, India

Mismatch (MMR) repair system plays a significant role in restoration of stability in the genome. Mutations in mismatch repair genes hamper their activity thus bring about a defect in mismatch repair (MMR) mechanism thereby conferring instability in the microsatellite sequences of both the coding and non-coding regions of the genome. Mutated mismatch repair genes result in the expansion or contraction of microsatellite sequence and confer microsatellite unstable or replication error positive phenotype. Hypermethylation of promoter regions of some of the MMR genes also causes inactivation of these genes and thus contribute to MSI. Microsatellite instability is an indicator of MMR deficiency and is a prime cause of varied tumorogenesis.

Papers

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 995-1000

Role of macrophage-colony stimulating factor and osteoclast differentiation factor in osteoclastogenesis of bone marrow derived stem cells

Mohamed Bayoumy, Uma Sankar & Natarajan Muthusamy

Received 6 May 2002; revised 3 June 2002

Macrophage colony stimulating factor (M-CSF) and osteoclast differentiation factor (ODF) regulate osteoclastogenesis in vivo. Regulation of osteoclast development in vitro by these cytokines has been reported in the present study. Simultaneous addition of ODF and M-CSF during initiation of bone marrow culture inhibited osteoclastogenesis. However, delayed addition of ODF (three days after initiation of the culture) resulted in dramatic increase in phenotypically and functionally mature osteoclast cells. Delayed addition of ODF beyond day three decreased osteoclastogenesis. Further, removal of M-CSF as early as day three inhibited ODF-induced osteoclastogenesis. These studies provided evidence for the importance of co-ordinated regulation of osteoclastogenesis by M-CSF and ODF.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1001-1004

Nuclear zinc in liver of cadmium and zinc treated rats

Ziad M Bataineh*

and

Jamil R Al-Alami

Received 21 November 2001; revised 7 June 2002

X-ray microanalysis was performed to detect quantitatively, the variation of the nuclear zinc in the liver cells of rats. The nuclear zinc concentration showed statistical decrease and increase in response to cadmium and zinc treatments, respectively. The results suggest that the liver responds differently to cadmium and zinc treatments. The difference in response to either treatment may reflect different mechanisms of zinc transport and metabolism in the liver. The difference in binding affinity of metallothionein (MT) may suggest the involvement of Mt in the metabolism and transport of zinc, an effect, which may be modified by treatment.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1005-1009

Antidiabetic and antioxidant effects of S-methyl cysteine sulfoxide isolated from onions (Allium cepa Linn) as compared to standard drugs in alloxan diabetic rats

Kumud Kumari& K T Augusti*

Received 26 February 2002; revised 25 June 2002

Antidiabetic and antoxidant effects of S-methyl cysteine sulfoxide (SMCS) isolated from A. cepa and two standard drugs, glibenclamide and insulin were studied and compared in alloxan diabetic rats after using each of them for treatment for two months. These drugs ameliorated the diabetic condition significantly, viz. maintenance of body weight and control of blood sugar in rats. Further they lowered the levels of malondialdehyde, hydroperoxide and conjugated dienes in tissues exhibiting antioxidant effect on lipid peroxidation in experimental diabetes. This is achieved by their stimulating effects on glucose utilization and the antioxidant enzymes, viz. superoxide dismutase and catalase. The probable mechanism of action of SMCS and glibenclamide may be partly dependent on the stimulation of insulin secretions and partly due to their individual actions. In the amelioration of diabetes the standard drugs showed a better action, but as an antioxidant SMCS proved to be a better one.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1010-1015

Effect of oxidative stress on serum and antioxidant enzymes in liver and kidney of rats and their modulation through dietary factors

H D Ramachandran, K Narasimhamurthy* & P L Raina

Received 13 December 2001; revised 13 May 2002

Modulatory effect of a formulated diet based on cereals, pulses and spices incorporated with crude palm oil (CPO), soybean oil (SBO) or cod liver oil (CLO) at 10% dietary level on oxidative stress and antioxidant enzymes was studied in liver and kidney tissues. Activity of alkaline phosphatase (ALP) and acid phosphatase (ACP) increased significantly in serum in various experimental groups. Significant increase in hepatic antioxidant enzymes, catalase, glutathione peroxidase (GPx) was also seen in the experimental groups. SOD activity showed a mixed response. Further, kidney antioxidant enzymes did not show much change compared to those in liver. The results indicated dietary lipid as the key players in determining cellular susceptibility to oxidative stress, which could be modulated by cereals, pulses and spices in the diet.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1016-1019

Response of liver antioxidant system to taurine in rats fed high fructose diet

A T Anitha Nandhini, S D Balakrishnan & C V Anuradha

Received 30 October 2001; revised 3 July 2002

Fructose-fed rats were more susceptible to peroxidative damage as measured by thiobarbituric acid reactive species. The concentrations of lipid peroxides, diene conjugates, lipofuscin and hydroperoxides were significantly higher. The levels of enzymic antioxidants such as vitamin C, vitamin E and glutathione and activities of antioxidant enzymes were significantly lower in fructose-fed rats. When these rats received taurine in drinking water, peroxidative damage was minimal in both plasma and liver. Taurine was effective in inducing the antioxidant potential in fructose-fed rats. Increased peroxidative damage in liver is likely to be associated with fructose dependent pathology, which could be reduced by taurine by enhancing the antioxidant potential.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1020-1025

Efficiency of Coleus aromaticus extract in modifying cyclophosphamide and
mitomycin-C induced clastogenicity in mouse bone marrow cells

Shyama Prasad S, Prashantha Naik & K K Vijayalaxmi

Received 6 September 2001; revised 15 May 2002

The anticlastogenic potency of the ethanolic extract of a medicinal plant, C. aromaticus was investigated by taking bone marrow chromosomal aberration assay andmicronucleus (MN) test as the test parameters. Swiss albino mice were fed orally with different doses (10,15, 25, 50 and 100 mg/kg body weight) of ethanolic extract for 7 days and on the 7th day, two doses each of anticancer drugs cyclophosphamide (CP; 25 and 50 mg/kg body weight) and mitomycin-C (MMC; 4 and 8 mg/kg body weight) were injected, ip, to different groups of animals. Bone marrow MN preparations were made at 24 and 48 hr time intervals. Coleus extract reduced CP and MMC induced MN and lower doses of the extract were found to be more effective than higher doses. The effective doses of extract in MN test were selected to study the anticlastogenic effects against CP (25 and 50 mg/kg body weight) and MMC (2 and 4 mg/ kg body weight) induced chromosomal aberrations. The results indicate the protective effect of C. aromaticus against CP and MMC induced cytogenetic damage.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1026-1031

Comparative immunopotentiating properties of saponin and incomplete
Freund’s adjuvant coupled to affinity purified larval antigen of
Hyalomma anatolicum anatolicum

S Ghosh, N K Singh & Preeti Rawat

Received 24 January 2002; revised 9 July 2002

Comparative immunostimulatory properties of saponin and incomplete Freund’s adjuvant (IFA) were studied using affinity purified 39 kDa larval antigen of Hyalomma anatolicum anatolicum. Significant antibody response to 39 kDa antigen was detected in the sera of rabbits immunized with both 39 kDa plus saponin and 39 kDa plus IFA in comparison to their corresponding control animals. Insignificant differences were noted in the antibody response between the animals of two immunized groups. Upon challenge, the animals immunized with 39 kDa plus IFA rejected 76.2±9.7% of larvae and 45.8±4.1% of adults while in group of animals injected with 39 kDa plus saponin rejected 80.9±11.2% of larvae and 47.2 ±5.7% of adults.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1032--1037

-fructofuranosidase production by 2-deoxyglucose resistant mutants of
Aspergillus niger in submerged and solid-state fermentation

Balasubramaniem Ashokkumar & Paramasamy Gunasekaran

Received 18 December 2001; revised 23 April 2002

Aspergillus niger produces extracellular β-fructofuranosidase under submerged (SmF) and solid state fermentation (SSF) conditions. After UV mutagenesis of conidiospores of A. niger, 2-deoxyglucose (10 g/l) resistant mutants were isolated on Czapek’s minimal medium containing glycerol as a carbon source and the mutants were examined for improved production of β-fructofuranosidase in SmF and SSF conditions. One of such mutant DGRA-1 overproduced β-fructofuranosidase in both SmF and SSF conditions. In SmF, the mutant DGRA-1 showed higher β-fructofuranosidase productivity (110.8 U/l/hr) than the wild type (48.3 U/l/hr). While in SSF the same strain produced 322 U/l/hr of β-fructofuranosidase, 2 times higher than that of wild type (154.2 U/l/hr). In SmF, both wild type and mutants produced relatively low level of β-fructofuranosidase in medium containing sucrose with glucose than from the sucrose medium. However in SSF, the DGRA-1 mutant grown in sucrose and sucrose+glucose did not show any difference with respect to β-fructofuranosidase production. These results indicate that the catabolite repression of β-fructofuranosidase synthesis is observed in SmF whereas in SSF such regulation was not prominent.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1038-1042

Infra-red spectroscopic analyses of banana waste degraded by oyster mushroom

G V Reddy, M P Shah, I L Kothari & A Ray

Received 4 May 2001; revised 11 April 2002

Carbon, hydrogen and nitrogen analyses of banana leaf and pseudostem biomass revealed their potentiality as substrates for microorganisms. Infra-red (IR) spectra of both biomass show presence of cellulose, xylan and lignin. IR spectra of leaf and pseudostem biomass degraded in solid state fermentation (SSF) by two Pleurotus species (P. sajor-caju and P. ostreatus) for 40 days showed the utilization of cellulose, xylan and lignin by these microbes. Dynamics of various lignocellulolytic enzymes of Pleurotus species and analyses of carbon, hydrogen and nitrogen contents of degraded biomass supported the same. Both the Pleurotus species exhibited lignin consumption ability on both the substrates.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1043-1049

A novel strategy of oxygen restriction by some diazotrophic enteric bacteria

V Kannan & P N Raju

Received 6 March 2002; revised 9 July 2002

Protection of nitrogenase against oxygen inactivation in diazotrophs involves numerous strategies. Glutathione is known to play an important role in scavenging oxyradicals in many living systems. The involvement of glutathione (reduced) (GSH), glutathione peroxidase (GPX) and glutathione reductase (GR) in the protection of nitrogenase in free living diazotrophs is reported here for the first time. Reduced glutathione content and the activity of glutathione peroxidase and glutathione reductase increased with increase in oxygen concentration under nitrogen fixing conditions but decreased under anaerobic and nitrogenase repressed conditions. This correlation is used to postulate a protecting role for GSH - GPX - GR system against oxygen inactivation of nitrogenase.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1050-1055

In vitro plantlet regeneration from seedling nodal explants of Acacia catechu

Rohini Sahni & Shrish C Gupta*

Received 25 September 2001; revised 22 May 2002

Multiple shoots were initiated after 20 days in stem nodes excised from in vitro grown seedlings of Acacia catechu, on Murashige and Skoog’s medium adjuvanted with 1 to 100 µM ofN6-benzyladenine (BA). Explants were subcultured on the same medium augmented with 1.5 g l-1 of polyvinylpyrrolidone (PVP) after 30 days. In the second subculture, after 30 days, the explants were transferred to a medium lacking PVP, but containing 10µM of BA, where nine or ten shoots differentiated per explant within next 30 days. If individual shoots along with some callus were subcultured on BA (10 µM), nearly 15 shoots per explant regenerated in 90 days. Thus, the average number of shoots obtained from each node was 142 after 180 days. Since a seedling develops four nodes after 20 days, theoretically an average of 568 shoots can be obtained from a single seed. If shoots were individually subcultured on ½-strength MS medium with 14.7 µM of indole-3-butyric acid (IBA), roots developed in 20 days. Addition of 40 mg l-1 of glutamic acid to the rooting medium prevented leaf senescence. These plantlets thrived well in garden soil, sand and silica (1:1:1).

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1056-1059

Esterase as molecular marker for salt tolerance in regenerated plants of rice,
Oryza sativa L

T S Swapna*

Received 6 September 2001; revised 23 May 2002

Esterase variation was studied in plants regenerated from callus cultures of four rice (Oryza sativa) varieties, viz. pokkali, which is a moderately salt tolerant variety and three salt sensitive varieties MI 48, annapoorna and jyothi. Variation was studied at tillering stage of plants regenerated from callus culture and germinated from seeds. Somaclonal variants for salt tolerance could be detected using variation in esterase banding pattern and activity.

Indian Journal of Experimental Biology

Vol. 40, September 2002, pp. 1060-1066

Bioregulation of carbohydrate metabolism in relation to source-sink operation
during grain-filling phase of growth in wheat

Parminder Sidhu & Rangil Singh*

Received 19 November 2001; revised 17 May 2002

Mobilization of free sugars from vegetative tissues to grain and their transformation to starch in relation to activities of some relevant enzymes during growth and development were investigated in wheat (Triticum aestivum L.). Vegetative tissues, viz. flag-leaf, flag-leaf sheath, nodes and internodes contained high concentration of free sugars from 70 DAS to 18 DPA and that was in the order of accumulation–flag-leaf sheathflag-leaf and internodesnodes. In these tissues, major portion of 14C appeared in endogenous sucrose, irrespective of the nature of [U-14C]-sugars supplied. In photosynthetic structures above flag-leaf node, namely peduncle, rachis and bracts, the free sugar make-up was maximum at anthesis (90 DAS). Activity of soluble acid invertase (EC 3.2.1.26) was high in these tissues during early stages of grain growth but reverse was true for soluble neutral invertase (EC 3.2.1.27) activity. In apical and basal portions of grain, free sugars were more or less similarly distributed in concentration. Linear and rapid accumulation of starch in endosperm paralleled with a decline in accumulation of this polymer in pericarp-aleurone. In the latter tissue, the activities of starch hydrolyzing enzymes, i.e - and -amylases (3.2.1.1 and 3.2.1.2) were high during initial stages of grain growth. During active grain-filling, alkaline inorganic pyrophosphatase (EC 3.6.1.1) seemed to play a vital role during starch accumulation in endosperm, whereas the involvement of 3-PGA phosphatase (EC 3.1.3.38) was almost confined to pericarp-aleurone. Impairement of ear head photosynthesis by shading depressed starch synthesis (~50%) indicating, thereby, the significant role of current photosynthates during grain-filling. The results suggested that grain growth in wheat was influenced by an efficient operation of source as well as regulatory factors, including enzymes, constituting intrinsic potential of grain sink.