Indian Journal of Experimental Biology

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VOLUME 45

NUMBER 3

MARCH 2007

CODEN : IJEB (A6) 45(3) 217-310(2007)

ISSN : 0019-5189

 

CONTENTS

 

Signaling molecules involved in the transition of growth to development of
Dictyostelium discoideum

223

      Hina A Mir, Jyotika Rajawat, Shalmali Pradhan & Rasheedunnisa Begum

 

 

 

Liposomes as adjuvant for combination vaccines

237

      Neeraj Mishra, Prem N Gupta, Sunil Mahor, Kapil Khatri, Amit K Goyal &
Suresh P Vyas

 

 

 

Effect of GnRH on guinea pig endometrium at preimplantation stage

242

      G A Jelodar, S Gholami & F Jafarpour

 

 

 

Remote ischaemic preconditioning and prevention of cerebral injury

247

      Ashish K Rehni, Richa Shri & Manjeet Singh

 

 

 

Circadian variation in lipid peroxidation induced by benzene in rats

253

      S V S Rana, Navita Chaudhary & Yeshvandra Verma

 

 

 

Paraffin baiting sytem for demonstration of growth and biofilm production in Pseudomonas aeruginosa

258

      Vishnu prasad S, Mamatha Ballal & P G Shivananda

 

 

 

Osmotic adjustment increases water uptake, remobilization of assimilates and maintains photosynthesis in chickpea under drought

261

      P S Basu, Masood Ali & S K Chaturvedi

 

 

 

Rapid micropropagation via axillary bud proliferation of Adhatoda vasica Nees from nodal segments

268

      Gauri Abhyankar & V D Reddy

 

 

 

Curcuma longa as feed additive in broiler bird and its patho-physiological effects

272

      P Kumari, M K Gupta, R Ranjan, K K Singh & R Yadava

 

 

 

Anti-inflammatory, antiarthritic and analgesic activity of a herbal formulation (DRF/AY/4012)

278

      Mahesh S Kaneria, S R Naik & R K Kohli

 

 

 

Characterization of antimicrobial compounds from a common fern, Pteris biaurita

285

      A K Dalli, G Saha & U Chakraborty

 

Effects of polyherbal formulation ‘ImmuPlus’ on immunity and disease resistance of Indian major carp, Labeo rohita at different stages of growth

291

      Jaya Kumari, P K Sahoo, & S S Giri

 

 

 

Preventive role of Gynandropsis gynandra L., against aflatoxin B1 induced lipid peroxidation and antioxidant defense mechanism in rat

299

      D Sivanesan & V Hazeena Begum

 

 

 

Hepatoprotective and antioxidant activities of flowers of Calotropis procera (Ait) R. Br. in CCl4 induced hepatic damage

304

      Absar Ahmed Qureshi, T Prakash, Tushar Patil, A H M Viswanath Swamy,
A Veeran Gouda, Prabhu K & S Ramachandra Setty

 

 

 

Author Index

Abhyankar Gauri

268

Ali Masood

261

Ballal Mamatha

258

Basu P S

261

Begum Rasheedunnisa

223

Begum V Hazeena

299

Chakraborty U

285

Chaturvedi S K

261

Chaudhary Navita

253

Dalli A K

285

Gholami S

242

Giri S S

291

Gouda A Veeran

304

Goyal Amit K

237

Gupta M K

272

Gupta Prem N

237

Jafarpour F

242

Jelodar G A

242

Kaneria Mahesh S

278

Khatri Kapil

237

Kohli R K

278

Kumari Jaya

291

Kumari P

272

Mahor Sunil

237

Mir Hina A

223

Mishra Neeraj

237

Naik S R

278

Patil Tushar

304

Prabhu K

304

Pradhan Shalmali

223

Prakash T

304

Qureshi Absar Ahmed

304

Rajawat Jyotika

223

Rana S V S

253

Ranjan R

272

Reddy V D

268

Rehni Ashish K

247

Saha G

285

Sahoo P K

291

Setty S Ramachandra

304

Shivananda P G

258

Shri Richa

247

Singh K K

272

Singh Manjeet

247

Sivanesan D

299

Verma Yeshvandra

253

Vishnu prasad S

258

Viswanath Swamy A H M

304

Vyas Suresh P

237

Yadava R

272

 

Keyword Index

ABLs

261

Adhatoda vasica

268

Adjuvants

237

Aeromonas hydrophila

291

AFB1

299

Analgesic effect

278

Antiarthritic

278

Anti-inflammatory

278

Antimicrobial compounds

285

Antioxidant

299, 304

Axillary bud

268

Benzene

253

Broiler birds

272

Calotropis procera

304

CCl4

304

Cerebral injury

247

Cicer arietinum

261

Circadian rhythm

253

Combination vaccines

237

Curcuma longa

272

Cytochrome P4502E1

253

D. discoideum

223

Diphtheria toxoid

237

F:C efficiency

272

Feed additive

272

Fern

285

GC-MS

285

GDT

223

GnRH

242

Growth

291

GSH

253

Guinea pig endometrium

242

Gynandropsis gynandra

299

 Hepatoprotection

304

Hexose

261

HPLC

285

Immunity

291

Immunostimulation              291

291

ImmuPlus (AquaImmu)

291

Ischaemic preconditioning

247

Labeo rohita

291

Leaf water potential

261

Lipid peroxidation

253, 259

Liposomes

237

Micropropagation

268

Nephroprotective effect

272

Nodal segments

268

Paraffin baiting

258

Photosynthesis

261

Preimplantation stage

242

Pseudomonas aeruginosa

258

Relative water content

261

Reperfusion

247

Signal transduction

223

Starch

261

Sucrose

261

Sucrose-phosphate synthase

261

Tetanus toxoid

237

Transpiration

261

Water stress

261

 

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 223-236

 

 

Review Article

 

Signaling molecules involved in the transition of growth to development of Dictyostelium discoideum

Hina A Mir, Jyotika Rajawat, Shalmali Pradhan & Rasheedunnisa Begum

 

The social amoeba Dictyostelium discoideum, a powerful paradigm provides clear insights into the regulation of growth and development. In addition to possessing complex individual cellular functions like a unicellular eukaryote, D. discoideum cells face the challenge of multicellular development. D. discoideum undergoes a relatively simple differentiation process mainly by cAMP mediated pathway. Despite this relative simplicity, the regulatory signaling pathways are as complex as those seen in metazoan development. However, the introduction of restriction-enzyme-mediated integration (REMI) technique to produce developmental gene knockouts has provided novel insights into the discovery of signaling molecules and their role in D. discoideum development. Cell cycle phase is an important aspect for differentiation of D. discoideum, as cells must reach a specific stage to enter into developmental phase and specific cell cycle regulators are involved in arresting growth phase genes and inducing the developmental genes. In this review, we present an overview of the signaling molecules involved in the regulation of growth to differentiation transition (GDT), molecular mechanism of early developmental events leading to generation of cAMP signal and components of cAMP relay system that operate in this paradigm.

 

 

Papers

Indian Journal of Experimental Biology 

Vol. 45, March 2007, pp. 237-241

 

 

Liposomes as adjuvant for combination vaccines

Neeraj Mishra, Prem N Gupta, Sunil Mahor, Kapil Khatri, Amit K Goyal & Suresh P Vyas

 

Received 17 March 2006; revised 18 October 2006

In the present study tetanus toxoid (TT) loaded liposomes and diphtheria toxoid (DT) loaded liposomes were prepared by reverse phase evaporation method and after combining these two vaccines the potential advantages were investigated. Prepared systems were characterized for the size, shape and entrapment efficiency. SDS-PAGE analysis of TT and DT was also performed. The selected liposomal formulations were administered subcutaneously to Balb/c mice and their immune responses were determined using ELISA after 15, 30, 45 days. After boosting the maximum immune response was observed after 45 days and was found to be 0.831 and 0.749 for TT loaded liposome and DT loaded liposomes respectively. When the mice were immunized subcutaneously with the physical mixture of TT loaded liposomes and DT loaded liposomes the immune response for the combination vaccine was found to be 1.44 and 0.741 for the TT and DT respectively. The result showed that the immune response of TT increased when it was combined with DT in liposomes. This confirms adjuvantcity of DT vis-a-vis immunogenicity. Thus, carrier mediated cocktail vaccination holds promise for clinical applications.

 

 

 

Indian Journal of Experimental Biology 

Vol. 45, March 2007, pp. 242-246

 

 

Effect of GnRH on guinea pig endometrium at preimplantation stage

G A Jelodar

 

and

S Gholami & F Jafarpour

 

Received 4 May 2006; revised 20 November 2006

Endometrium of GnRH treated group resembled with pregnant group and endometrial thickness in these groups significantly increased in comparison with non-pregnant group.  In GnRH treated animals, most of histomorphological changes in epithelial cells, glands and stroma of uterus was similar to pregnant group. The results revealed that mammalian form of GnRH exerted endometrial change in guinea pig almost similar to those occur in normal pregnant animals and its administration prior to implantation may improve pregnancy rate following embryo transfer.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 247-252

 

 

Remote ischaemic preconditioning and prevention of cerebral injury

Ashish K Rehni, Richa Shri & Manjeet Singh

 

Received 20 September 2005; revised 9 October 2006

Bilateral carotid artery occlusion of 10 min followed by reperfusion for 24 hr was employed in present study to produce ischaemia and reperfusion induced cerebral injury in mice. Cerebral infarct size was measured using triphenyltetrazolium chloride staining. Short-term memory was evaluated using elevated plus maze. Inclined beam walking test was employed to assess motor incoordination. Bilateral carotid artery occlusion followed by reperfusion produced cerebral infarction and impaired short-term memory, motor co-ordination and lateral push response. A preceding episode of mesenteric artery occlusion for 15 min and reperfusion of 15 min (remote mesenteric ischaemic preconditioning) prevented markedly ischaemia-reperfusion-induced cerebral injury measured in terms of infarct size, loss of short-term memory, motor coordination and lateral push response. Glibenclamide (5 mg/kg, iv) a KATP channel blocker and caffeine (7 mg/kg, iv) an adenosine receptor blocker attenuated the neuroprotective effect of remote mesenteric ischaemic preconditioning. It may be concluded that neuroprotective effect of remote mesenteric ischaemic preconditioning may be due to activation of adenosine receptors and consequent activation of KATP channels in mice.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 253-257

 

 

Circadian variation in lipid peroxidation induced by benzene in rats

S V S Rana, Navita Chaudhary & Yeshvandra Verma

 

Received 8 August 2005; revised 16 December 2006

Time-dependent effect of benzene, a potent carcinogenic industrial solvent, on lipid peroxidaiton and associated mechanisms has been studied in liver and kidney of rats. Significant differences were observed in the values of urinary phenol, microsomal malondialdehyde, reduced glutathione (GSH) and cytochrome P4502E1 in rats treated with benzene in morning and evening hours. Higher were the values for urinary phenol and hepatic microsomal malondialdehyde in rats administered benzene in evening hours. Contrarily, higher were the values for GSH and cytochrome P4502E1 in rats treated with benzene in morning hours. Increased microsomal lipid peroxidation has been attributed to low GSH status, whereas increased phenol concentration could be related to low activity of cytochrome P4502E1 in the liver of rats in evening hours. It is concluded that circadian rhythmicity in hepatic drug metabolizing enzyme system and GSH contributes in toxicity of benzene. The results are important from occupational health point of view.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 258-260

 

 

Paraffin baiting system for demonstration of growth and biofilm production in Pseudomonas aeruginosa

 

Received 14 June 2006; revised 16 October 2006

Pseudomonas aeruginosa is one of the commonest pathogens among the pseudomonads. This organism can grow in minimal nutritional requirements. Because of the ability of pseudomonads to grow on paraffin is not commonly found among other human pathogens and the primary human pathogen being P. aeruginosa, we studied the adaptation of this organism to paraffin baiting system for growth and biofilm formation. Strains were tested for the capacity to use paraffin as the sole source of carbon using Czapek’s minimal salt medium. Of the 53 clinical isolates of P. aeruginosa,
20 strains exhibited growth by 24 hrs and 42 strains by 48 hrs. The remaining strains did not show any growth in the paraffin baiting system. The oxidase test with the paraffin baiting system was also performed. This simple and inexpensive method can be used to isolate and demonstrate the biochemical and biofilm forming capacity of the organism.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 261-267

 

 

Osmotic adjustment increases water uptake, remobilization of assimilates and maintains photosynthesis in chickpea under drought

P S Basu, Masood Ali & S K Chaturvedi

 

Received 25 July 2006; revised 10 October 2006

Eight chickpea advanced breeding lines (ABLs) and their parents were evaluated for osmotic adjustment (OA), leaf carbohydrates and gas exchange under dryland field . These (ABLs) were derived from crosses between CTS 60543 ´ Kaniva and Tyson ´ Kaniva. Mean leaf water potential (LWP) fell down from –1.00 MPa at pre-stress level to about –2.25 MPa during terminal stress. Relative water content (RWC) showed periodic changes with alternate decrease or increase at certain interval, which also influenced the values of OA (low or high) in number of genotypes e.g. Kaniva, CTS 60543, Tyson and M 75. Significant variation in OA ranging 0.45 to 0.88 MPa was observed at high level of stress at –2.5 MPa. However, none of the genotypes showed stability of OA over the period of stress. Leaf starch declined even at mild stress (LWP,-1.6 MPa) resulting in an increase in hexose sugars and activation state of sucrose-phosphate synthase (SPS) that led to accumulation of  sucrose. Both photosynthesis (Pmax) and transpiration decreased concurrently in two chickpea lines M 129 and Tyson with increasing water stress. However, rate of decline in the photosynthesis slowed down even drought was further intensified. The observed periodic changes in OA, RWC and photosynthesis appeared to be associated with drought-induced changes in SPS and carbohydrates which modify water uptake of the leaves.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 268-271

 

 

Rapid micropropagation via axillary bud proliferation of Adhatoda vasica Nees from nodal segments

Gauri Abhyankar & V D Reddy

 

Received 16 January 2006; revised 10 October 2006

A protocol for rapid multiplication of Adhatoda vasica has been developed through nodal explants from field grown mature plants. The maximum number of shoots, i.e., 7.75 + 0.392 differentiated from split nodal halves on MS medium supplemented with BA (10.0 mg/l) during 4 weeks of culture. Maximum number of shoots formed per explant increased to ca. 30 within 6 weeks of subculture on medium containing BA (1.0 mg/l) and Kn (1.0 mg/l). The isolated shoots rooted 90% in MS medium containing IBA (0.1 mg/l) in 2 weeks. The rooted plantlets were successfully transferred to soil in glasshouse and subsequently in field. The plantlets rooted in liquid medium did not survive, but those rooted on solid medium showed more than 75% survival. In vitro raised plants grew successfully ex vitro till flowering.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 272-277

 

 

Curcuma longa as feed additive in broiler birds and its patho-physiological effects

P Kumari, M K Gupta, R Ranjan, K K Singh & R Yadava

 

Received 7 March 2006; revised 30 August 2006

Broiler birds (Vencob chicken of 3 days old) when given feed mixed with powdered rhizome of Curcuma longa
(CL; @ 1 g/kg) for 42 days of age, showed significant decrease in serum uric acid and albumin as compared to control, whereas significant increase was recorded in the level of serum total protein and globulin. Level of serum glucose, alkaline phosphatase, aspartate amino transferase and calcium showed no significant variation between the two groups. Micronutrient assay revealed significantly higher level of manganese, zinc, iron and copper in treated group as compared to control group. HA/HI test revealed better humoral response against RD vaccine in CL administered birds. Haematological study showed significantly higher haemoglobin and absolute neutrophil count in treated group. Addition of CL as feed additive also resulted in better growth rate, feed consumption and F:C efficiency in the treated birds. Thus, it could be concluded that powdered CL might be a useful feed additive, since it enhanced the F:C efficiency and had nephroprotective properties.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 278-284

 

 

Anti-inflammatory, antiarthritic and analgesic activity of a herbal
formulation (DRF/AY/4012)

Mahesh S Kaneria1, S R Naik & R K Kohli

 

Received 20 January, 2006; revised 7 October 2006

Anti-inflammatory, antiarthritic and analgesic effect of a herbal product (DRF/AY/4012) was evaluated in animal models. Herbal product treatment induced a dose dependent anti-inflammatory activity in acute inflammatory models (carrageenin and egg–albumin induced rat hind paw edema). It also elicited promising anti-inflammatory activity in chronic inflammatory models (cotton pellet granuloma and Freund’s adjuvant induced polyarthritis in rats). Further, the product inhibited the increased level of serum lysosomal enzyme activity viz. serum glutamic oxaloacetic transaminase, serum glutamic pyruvic transaminase, alkaline phosphatase and the lipid peroxidation in liver. In Freund’s adjuvant induced polyarthritis, herbal product reduced the increased level of hydroxy proline, hexosamine and total protein content in edematous tissue .The  product also exhibited mild to moderate analgesic activity in acetic acid induced writhing in mice. The LD50 value of the herbal product was more than 16gm/kg by oral route in mice. The product has distinct advantages over the existing agents and deserves further developmental studies.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 285-290

 

 

Characterization of antimicrobial compounds from a common fern, Pteris biaurita

A K Dalli, G Saha & U Chakraborty

 

Received 29 May 2006; revised 5 December 2006

Methanol extract was prepared from the fronds of Pteris biaurita and partial purification was done by solvent partitioning with diethyl ether and ethyl acetate, followed by hydrolysis and further partitioning with ethyl acetate. The three fractions, thus obtained were bioassayed separately against five test fungi- Curvularia lunata, Fomes lamaoensis, Poria hypobrumea, Fuasrium oxysporum and a bacterium- Bacillus pumilus, by spore germination, radial growth and agar cup techniques. Results revealed that ethyl acetate fraction (III) contained the active principle. TLC plate bioassay of the active fraction revealed inhibition zone at an Rf of 0.5-0.65. Silica gel from this region was scraped, eluted in methanol and subjected to UV-spectrophotometric analysis. An absorption maxima of 278 nm was recorded. HPLC analysis of TLC-eluate revealed a single peak with retention time of 8.1 min. GC-MS analysis revealed six major peaks in the retention time range of 7.2-10.9 min. Comparison with GC-MS libraries revealed that the extracts may contain a mixture of eicosenes and heptadecanes.

 

 

 

Indian Journal of Experimental BiologyVol. 45, March 2007, pp. 291-298

 

 

Effects of polyherbal formulation ‘ImmuPlus’ on immunity and disease resistance of Indian major carp, Labeo rohita at different stages of growth

Jaya Kumari, P K Sahoo & S S Giri

 

Received 6 June 2005; revised 14 July 2006

A series of experiments were performed to determine the impact of polyherbal immunomodulatory formulation ‘ImmuPlus (AquaImmu) on growth, immunity and disease resistance of rohu (Labeo rohita), one of the Indian major carp at different stages of growth. Rohu larvae were fed on plankton, ImmuPlus-mixed compound feed, and plankton plus ImmuPlus-mixed compound feed (ImmuPlus added at three dose levels of 0.25, 0.50, and 0.75 g/kg feed) from 4th day of hatching to 14th day. ImmuPlus-mixed diets enhanced growth of larvae, survival and disease resistance against Aeromonas hydrophila challenge, compared to only plankton-fed group. In two other experiments, advanced rohu larvae and fingerlings were fed with ImmuPlus-mixed compound feed (at 0, 0.5, 1.0 and 2.0 g/kg) for 15, 30 and 45 days. At the end of 45 days for advanced larvae and 30 days for fingerlings, the fish fed with ImmuPlus at 1.0 g/kg level showed significantly higher growth and disease resistance against A. hydrophila challenge. In a separate experiment, juveniles of rohu were fed with 1 g/kg of ImmuPlus incorporated feed for 15 and 30 days. At the end of the trial, the ImmuPlus fed fish showed enhanced non-specific immunity (as measured through nitroblue tetrazolium reduction assay, serum lysozyme activity, serum haemolysin titre and resistance against A. hydrophila challenge in non-vaccinated fish as well as specific immunity levels (as measured through bacterial agglutination titre against A. hydrophila in vaccinated fish). Incorporation of ImmuPlus at 1 g/kg level in the diet of rohu may be beneficial for enhancing disease resistance.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 299-303

 

 

Preventive role of Gynandropsis gynandra L., against aflatoxin B1 induced lipid peroxidation and antioxidant defense mechanism in rat

D Sivanesan & V Hazeena Begum

 

Received 24 March 2006; revised 28 September 2006

G.gynandra extract was found to potentially diminish the rate of lipid peroxidation, with a significant increase in the levels of enzymatic (superoxide dismutase, catalase and glutathione peroxidase) and non-enzymatic (reduced glutathione vitamins E and C, and uric acid) antioxidants, which were found, altered during aflatoxin B1 (AFB1) injection. The result confirmed that G.gynandra extract exerts its chemopreventive efficacy by preventing the rate of lipid peroxidation and influenced the enzymatic and non-enzymatic antioxidants in AFB1 induced male albino rats.

 

 

 

Indian Journal of Experimental Biology

Vol. 45, March 2007, pp. 304-310

 

 

Hepatoprotective and antioxidant activities of flowers of Calotropis procera (Ait) R. Br. in CCl4 induced hepatic damage

Absar Ahmed Qureshi, T Prakash, Tushar Patil, A H M Viswanath Swamy, A Veeran Gouda,
Prabhu K & S Ramachandra Setty

 

Received 22 February 2006; revised 16 October 2006

Hepatoprotective activity of 70% ethanolic extract of flowers of C. procera was studied against CCl4 induced hepatic injury in albino rats and mice. In addition, antioxidant activity was studied by in vitro models. Pre-treatment with 70% ethanolic extract (CPA) reduced the biochemical markers of hepatic injury like serum glutamate pyruvate transaminase, serum glutamate oxaloacetate transaminase, alkaline phosphatase, bilirubin, cholesterol, HDL and tissue glutathione (GSH) levels. Similarly pretreatment with CPA reduced the CCl4 induced elevation in the pentobarbitone sleeping time. Histopathological observations also revealed that pretreatment with CPA protected the animals from CCl4 induced liver damage. CPA demonstrated dose dependant reduction in the in vitro and in vivo lipid peroxidation induced by CCl4. In addition it showed dose dependant free radical scavenging activity. The results indicate that flowers of C. procera possess hepatoprotective property possibly because of its anti-oxidant activity. This property may be attributed to the quercetin related flavonoids present in the flowers of Calotropis procera.