Indian Journal of Experimental Biology

CONTENTS

Indian Journal of Experimental Biology

 Vol 42, November 2004, pp. 1043-1055

 Review Articles

Fertilization, embryonic development and oviductal environment: Role of estrogen induced oviductal glycoprotein

Purvi Bhatt, Kaushiki Kadam, Anjana Saxena & Usha Natraj

Mammalian oviduct is the physiological site for sperm capacitation, gamete fertilization and early embryonic development. The secretory cells lining the lumen of the mammalian oviduct synthesize and secrete high molecular weight glycoprotein (OGP) in response to estrogen. The protein has been shown to interact with gametes and early embryo. Several key functions have been postulated particularly its role in pre-implantation events which would have far reaching implications in assisted reproductive technology and in the development of non-hormonal contraceptive vaccine. The intention of this article is to discuss the current status of the protein and analyze how far the postulated function of OGP has been borne out by the available data.

Indian Journal of Experimental Biology

 Vol 42, November 2004, pp. 1056-1065

Enhancement of gastrointestinal absorption of poorly water soluble drugs via lipid based systems

Natesan Subramanian & Saroj K Ghosal

Development of knowledge on lipids has attracted the scientific community for the effective utilization of the natural and synthetic lipids. Bioavailability of poorly water soluble drugs from gastrointestinal tract (GIT) can be enhanced by formulating the drugs in lipid based formulations. This formulation can increase the dissolution of poorly water soluble drugs, and facilitates the formation of solubilized phases from which absorption may occur. The enhanced solubility of lipophilic drugs from lipid-based systems will not necessarily arise directly from the administered lipid, but most likely from the intra luminal processing to which they are subjected prior to absorption. This review will focus on assessment of lipid-based formulations of drugs with a consideration of how gastrointestinal physiology, the choice of lipids and their formulation attribute and the mode of lipid digestion in the GIT influence the bioavailability of lipophilic drugs.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1066-1072

Papers

Synthesis, biological evaluation and kinetic studies of glyceride prodrugs of diclofenac

M S Y Khan & Mymoona Akhter

The prodrugs (glyceride derivatives) 3a and 3b of diclofenac were prepared by reacting 1, 2, 3-trihydroxy propane- 1,3-dipalmitate/stearate with the acid chloride of diclofenac as potential prodrugs to reduce the gastrointestinal toxicity associated with them. These prodrugs were evaluated for their ulcerogenicity, anti-inflammatory and analgesic activity. It was found that the prodrugs were significantly less irritating to the gastric mucosa as indicated by severity index of 0.86, 0.78 compared to 1.6 of diclofenac. The prodrugs 3a and 3b showed better anti-inflammatory and analgesic activity than the parent drugs. The hydrolysis of prodrugs 3a and 3b were studied at pH 3, 4, 5 and 7.4. The HPLC analysis showed that the prodrugs were resistant to hydrolysis at pH 3, 4 and 5 indicating that they did not hydrolyze in acidic environment, whereas at pH 7.4 the prodrugs readily released the parent drug in significant quantities. The plasma levels of diclofenac were also analyzed by HPLC in rats after single oral dose of the prodrugs. The results indicated that the parent drugs were readily released. The concentration of diclofenac during the study was found higher in animals treated with prodrugs 3a and 3b compared with animals treated with diclofenac. The concentration of diclofenac was found to be 38.59, 33.6 and 30.36 μg/ml in animals treated with prodrugs 3a, 3b and diclofenac respectively. In conclusion, all these studies indicated that the glyceride prodrugs of diclofenac might be considered as potential biolabile prodrugs of diclofenac.

Indian Journal of Experimental Biology

Vol. 42, November 2004, pp. 1073-1077

Immunomodulation by ‘ImmuPlus (AquaImmu)’ in giant freshwater prawn, Macrobrachium rosenbergii (De Man)

Jaya Kumari, P K Sahoo, S S Giri & Bindu R Pillai

ImmuPlus, a polyherbal commercial formulation was used to modulate the immune system of commercially important giant freshwater prawn M. rosenbergii. The prawns were fed with basal diet supplemented with ImmuPlus @ 1g/kg feed for 4 weeks. Results showed that the phenoloxidase activity (PO), haemagglutination and lysozyme activities were significantly elevated in ImmuPlus-fed prawn up to 3 weeks of feeding and declined after 4 weeks of feeding. The total protein level in ImmuPlus-fed prawn raised up to 2^nd week of feeding. Incorporation of ImmuPlus at the rate of 1g/kg feed in the diet of prawn for 3 weeks may be beneficial in raising the immune status of prawn.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1078-1083

Induction of chromosomal aberrations and sister chromatid exchanges by chlormadinone acetate in human lymphocytes: A possible role of reactive oxygen species

Yasir Hasan Siddique & Mohammad Afzal

Genotoxicity study of a synthetic progestin chlormadinone acetate (CMA) was carried out in human lymphocytes using chromosomal aberrations (CAs) and sister chromatid exchanges (SCEs) as parameter. Effect of CMA was studied at 10, 20, 30 and 40 mM. CMA was genotoxic at 30 and 40 mM. With a view to study the possible mechanism of genotoxicity of CMA, superoxide dismutase (SOD) and catalase (CAT) were used separately and in combination along with the CMA (40 mM) at different doses. SOD treatment increased CAs and SCEs at both the doses. CAT treatment decreased the frequencies of CAs and SCEs in both, separately and in combination with SOD, suggesting a possible role of reactive oxygen species for the genotoxic damage.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1084-1090

Ovarian monosaccharides (glucose and fructose): Hormonal effects and their role in final oocyte maturation and egg quality in catfish Heteropneustes fossilis, Bloch.

A Mishra & K P Joy

In the catfish H. fossilis, ovarian glucose and fructose levels showed significant annual variations that could be correlated with the gonado-somatic index. The concentration of the sugars, which was low in resting phase of gonad increased with the onset of oogenesis, reaching the peak in prespawning (June) phase. Glucose is the predominant monosaccharide in the late oogenesis, while fructose in early oogenesis and resting phase. Administration of a single injection of HCG (100 IU/fish) decreased glucose at 4 hr but increased fructose (6 hr) in the resting phase. In the preparatory phase (active oogenesis), fructose decreased significantly at 4 hr and glucose at 6 and 12 hr. In the prespawning phase, HCG administration induced a significant decrease in glucose level at 2 hr but increased it significantly at 16 and 18 hr. Fructose concentration, on the other hand, decreased ≥ 4 hr. The involvement of different energy substrates (glucose, fructose, 2-deoxyglucose and pyruvate) on final oocyte maturation and viability of eggs were demonstrated in an in vitro study, employing incubation of mature oocytes in medium containing one or the other energy substrates. Glucose at 4 and 8 mM concentrations increased the viability of eggs without affecting germinal vesicle break down (GVBD) and at high concentrations (16 and 32 mM) marginally increased GVBD but decreased viability. Pyruvate at high concentrations (≥ 8 mM) produced a positive effect on both GVBD and viability. Fructose and 2-deoxyglucose did not elicit any effect on these parameters. The results suggest that glucose has an active role in catfish oogenesis and maintaining egg quality and HCG influences monosaccharide metabolism in a season-dependent manner.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1091-1095

Ultrastructural changes in cauda epididymidal epithelial cell types of Azadirachta indica leaf treated rats

M G Ghodesawar, r Nazeer ahamed, A W Mukhtar Ahmed & R H Aladakatti

To assess if cauda epididymis is a target for the effect of A. indica leaves, Wistar strain male albino rats were administered (po) A. indica leaves (100 mg/rat/day for 24 days). Transmission electron microscopic analysis revealed that in the cauda epididymal epithelium the nuclei of principal cells were enlarged and the number of coated micropinocytotic vesicles of the apical cytoplasm decreased. Microvilli were missing and mitochondrial cristae and Golgi complex were highly disrupted. The cytoplasm was abounding with lysosomal bodies. The clear cells increased in perimeter and their nuclei increased in size and contained lesser chromatin. The nuclear membrane bulged out. The cytoplasm was vacuolized. Further, there was decrease in size of the lipid droplets, mitochondria, Golgi complex, endoplasmic reticulum and there was accumulation of lysosomal bodies. The changes in the principal and clear cells appear to be due to the effect of the hypoandrogen status caused by treatment with A. indica leaves and a direct action on the epididymal epithelium.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1096-1099

Phostoxin–induced biochemical and pathomorphological changes in rabbits

Ngozi Paulinus Okolie, Jonathan Umezuluike Aligbe & Edorisiagbon Eguagu Osakue

The effect of chronic phostoxin administration on some tissue ATPases, hematology and tissue histopathology was investigated using a combination of gravimetric, enzymatic, colorimetric and histological procedures in New Zealand White rabbits after 2 weeks administration of 0.8mg phostoxin/kg body weight/day, po. The phostoxin treatment led to significant decreases in Na⁺–K⁺ ATPase activities in renal, hepatic and cardiac tissues. Similar decreases were obtained in the activities of Ca²⁺–ATPase and Mg²⁺–ATPase in liver. In addition, the phostoxin-toxified rabbits manifested significant decreases in hematocrit, red blood cell count, hemoglobin and platelets. Histological examination of the tissues revealed pronounced degenerative changes in liver, heart and kidney.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1100-1106

Pathogenic effects of Salmonella enterica subspecies enterica serovar Typhimurium on sprouting and growth of maize

B R Singh, Ravikant Agarwal & Mudit Chandra

The study was undertaken to understand effects and survival of S. enterica subspecies enterica serovar Typhimurium (S. Typhimurium), a zoonotic serovar, on maize seed germination and plant growth. All the four strains of S. enterica subspecies enterica serovar Typhimurium significantly reduced germination of maize seeds in sprouting plates as well as in soil. About ≥2.7´10³ Salmonella cfu ml^-1 of soaking water, while ≥2.7´10⁷ Salmonella cfu g^-1 soil were required to significantly inhibit germination of maize. Similar inhibition of germination could be observed using ≥ 16 mg of bacteria free Salmonella cell lysate (CL) protein per g of soil or ≥0.5 mg of CL protein per ml of soaking water in sprouting plates. At the constant dose of 3.6´10⁷ to 3.8´10⁷ Salmonella cfu or 5 mg cell lysate protein ml^-1 of soaking water, four strains of Salmonella significantly reduced germination, however difference between strains was insignificant. After germination too, maize growth was affected both by Salmonella organism and CL with little strain-to-strain variation. All Salmonella persisted in growing plants from 15 to 35 days of plant age and up to 190 days in soil. Maize plants once grown for a week in sterile soil were resistant to invasion of S. enterica subspecies enterica serovar Typhimurium in their leaves even in doses as high as 7.6´10⁹ cfu g^-1 of soil. Salmonella persisted better and longer in plants grown from contaminated seed sown in loam soil, but rarely in plants grew in sandy soil. All maize plants had Salmonella in their stumps even after 35 days of sowing irrespective of kind of soil, primary source of infection (soil or seed) and type of S. enterica subspecies enterica serovar Typhimurium strain. The study revealed that Salmonella is not only zoonotic but a phytopathogen also.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1107-1111

Pathology of benzalkonium chloride toxicity and its effect on body weight gain in broiler birds

P K Mandal, M K Gupta & K K Singh

Four groups comprising 16 broiler birds each were given benzalkonium chloride (BC) at 100, 300, 500 and 700 ppm in drinking water for 40 days and one group of 16 birds (control) was kept on plain water. Clinical signs in higher dose groups were respiratory distress, drooling of saliva, difficulty in deglutition, inappetence, apathy, lethargy and loss of body weight. Better body weight gain was recorded in 100 ppm dose rate. At 300 ppm, no significant body weight variation was recorded, whereas, at 500 and 700 ppm dose rates, significantly poor body weight gain was recorded. Major pathological changes were seen in 500 and 700 ppm groups, which exhibited formation of yellow diphtheritic plaques in the buccal cavity, swollen and pale commissures of beak and shortening of tongue. Minute necrotic and ulcerative foci were seen in oesophagus and crop. Hyperplastic and hypertrophic alterations were seen in mucosa of the upper digestive tract. Crop of 300 ppm group revealed formation of well developed epithelial nest with pseudoepitheliomatous hyperplasia at the margin of the lesion. Serum alanine transaminase, urea and uric acid in 500 and 700 ppm groups were elevated whereas no significant variations were observed in the 100 and 300 ppm groups. BC could enhance performance of broiler birds at 100 ppm dose rate. It should not be used beyond 300 ppm.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1112-1116

Occurrence of chromium resistant thermotolerant coliforms in tannery effluent

Tuhina Verma, Pramod W Ramteke & Satyendra K Garg

Twenty six thermotolerant strains resistant to high levels of chromium (50-250 µg/ml) were isolated from treated tannery effluent. They were also found resistant to multiple heavy metals and antibiotics. Majority of them were resistant to copper and bacitracin. Nine strains representing different resistance patterns were selected for plasmid profile and conjugation studies. Agarose gel electrophoresis results revealed that 6 strains harboured a single plasmid, whereas 3 strains exhibited 2 plasmid bands. Among antimicrobials, co-trimazole and bacitracin and among metals, Cu²⁺, Cd²⁺, Zn²⁺ and Ni²⁺ resistance were transferred most frequently at variable rates. However, chromium resistance was transferred in 6 strains with a frequency ranging 19-49´10^-2. Resistance to Co²⁺ and Hg²⁺ did not transfer under environmental conditions. Among the nine strains, three were found predominantly uropathogenic Escherichia coli (UPEC) serotype 04, whereas two strains were untypable. In addition, 4 transconjugants also showed a positive result after serotyping.

Indian Journal of Experimental Biology

Vol. 42, November 2004, pp. 1117-1122

Mechanism of nickel resistance in a cobalt-resistant wall-less mutant of Neurospora crassa (fz; sg; os-1)

R Sri Rajyalaxmi, T Naga Sowjanya, P Kiranmayi & P Maruthi Mohan

A cobalt-resistant wall-less mutant of N. crassa (Cor-sl) characterized previously was also found to be 3-fold more resistant to nickel when compared to the parent wall-less mutant (W-sl). The Cor-sl strain accumulates relatively lower amounts of nickel when compared to W-sl. Sub-cellular fractionation showed significant quantities of nickel to be associated with nuclear and mitochondrial fractions in both the wall-less mutants. However significant differences were observed in vacuolar fractions of W-sl and Cor-sl strains. Fractionation of cell-free extracts on .Sephadex G-10 column resolved nickel into two peaks, of which the peak II in Cor-sl constituted 70% of nickel, while the same in W -sl was about 30%. A 3-fold increase in histidine content was observed in case of Cor-sl as compared to W-sl strain, suggesting its role in Ni-resistance.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1123-1131

Formulation of medium constituents by multi response  analysis of central composite design to enhance chitinase production in Pantoea dispersa

Vipul Gohel, Smita Trivedi, Pranav Vyas & H S Chhatpar

Received 26 April 2004; revised 9 July 2004

In the present study, a high chitinase producing strain Pantoea dispersa was isolated from the sea dumps at Bhavnagar, India. Chitin, urea, CaCl₂ and MgSO₄^.7H₂O were variables used in central composite design for chitinase production. Chitinase, biomass and pH were the responses used in different models to evaluate individually fit ones. Quadratic model was found to be fit for chitinase response whereas in the case of biomass and pH, linear model was found to be fit without the effect of others. Chitinase production was optimized with respect to other responses such as biomass and pH in multiresponse analysis of response surface design by using desirability approach. In multiresponse analysis, following medium formulation (g/l), chitin, 15; urea, 0.32; CaCl₂, 0.10 and MgSO₄.7H₂O, 0.08 was found to predict optimum chitinase production of 482.77 units/ml with overall highest desirability of 0.854 as compared to other formulations. The selection of model was done on the basis of high Adjusted R-squared value and lowered p-value for each model in individual analysis of each response. In multiresponse experiment, it was found that for response chitinase quadratic model and for responses pH and biomass linear models were well fit. Through desirability analysis, it was found that in the chitinase production, pH was essential as compared to biomass in P. dispersa. Endochitinase and chitobiase actvities were also studied.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1132-1135

Evaluation, grafting success and field establishment of cashew rootstock as influenced by VAM fungi

R Lakshmipathy, A N Balakrishna, D J Bagyaraj, D A Sumana & D P Kumar

Seven isolates of vesicular arbuscular mycorrhizal (VAM) fungi were isolated from cashew rhizosphere soil of different cashew growing regions of South India. These seven isolates along with two more VAM fungi namely Acaulospora laevis and Glomus mosseae, which were found to be better symbionts for cashew during our earlier study were used to study their effectiveness on the growth and nutrition of cashew rootstock Ullal-1. Four promising VAM fungi were selected based on this study. Rootstocks inoculated with these four fungi were evaluated for their vigour through grafting success, using Ullal-3 cashew variety as scion. Grafting success was more in rootstocks inoculated with A. laevis and one of local isolates Glomus etunicatum. Grafts with rootstock treated with G. etunicatum and A. laevis survived and performed better when planted in the field compared to the uninoculated and other VAM fungal treatments.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1136-1140

In vitro clonal propagation of bird eye chilli (Capsicum frutescens Mill.)

H B Gururaj, P Giridhar, Ashwani Sharma, B C N Prasad & G A Ravishankar

An efficient and highly reproducible protocol for micropropagation of bird eye chilli Capsicum frutescens was attempted. Murashige and Skoog (MS) medium containing 0.5-3.0 mgl^-1 of 6-benzyladenine (BA), 2-isopentenyl adenine (2iP), kinetin and 0.5-2.0 mg l^-1 of indole-3-acetic acid (IAA), indole-3-butyric acid (IBA) along with 1 gl^-1 activated charcoal (AC) were used for shoot regeneration from both shoot tip and nodal explants. Shoot tip explants (100%) grew well on medium containing 1 mgl^-1 of kinetin and 1 mgl^-1 of IBA. Shoot proliferation (1-3) from nodal explants was effective on this medium. The regenerated shoots with 4-7 nodes had further growth upon sub-culturing onto kinetin (1 mgl^-1) and IBA (1 mgl^-1) and rooted simultaneously. The rooted plants were transferred to pots after hardening under controlled conditions. The survival percentage in pots was 80-90%.

Indian Journal of Experimental Biology

 Vol. 42, November 2004, pp. 1141-1146

Notes

Pre-electroconvulsive shock administration of calcium channel blockers reduces retrograde amnesia induced by ECS

M Sushma, S Sudha & S Guido

Effect of pre-electroconvulsive shock (ECS) administration of calcium channel blockers (CCBs) like verapamil, diltiazem, nifedipine, nimodipine, flunarizine and cinnarizine on retrograde amnesia induced by ECS was examined using passive avoidance paradigm in rats. The groups (Gr 1-7) of adult, male Wistar rats received true ECS with CCBs (5mg/kg; i.p) or vehicle (10 ml/kg; ip) and other groups (Gr 8-14) received sham ECS with CCBs (5mg/kg; i.p) or vehicle (10 ml/kg; i.p). The anti-amnestic activity of CCBs were evaluated using the passive avoidance paradigm in rats. Results showed that, the baseline latencies for all the groups did not differ significantly. Rats receiving true ECS produced significantly lower latencies. There was increase in the post ECS step through latencies of the rats administered CCBs before ECS. Therefore, pre-ECS administration of calcium channel blockers might reduce retrograde amnesia produced by ECS without altering seizure duration.

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