Indian Journal of Biochemistry 

& Biophysics

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CONTENTS

 

Papers
Studies on surfactant-biopolymer interaction. II. Interaction of cetyl trimethyl ammonium-, cetyl ethanolyl dimethyl ammonium-, cetyl diethanolyl methyl ammonium-, and cetyl triphenyl phosphonium bromides and cetyl pyridinium chloride with calf thymus DNA	205
Arindam Chatterjee and Satya P Moulik*
Oxidative stress in tumour-bearing fore-stomach and distant normal organs of Swiss albino mice	216
Gagandeep, A R Rao and R K Kale*
Induction of apoptosis in pheochromocytoma (PC12) cells exposed to eicosapentaenoic acid in vitro	222
Mei Li, Haitao Ge, Xiuqin Kong, Weifa Zheng and Zhili Liu*
Molecular and biochemical markers associated with leaffolder (Cnaphalocrocis medinalis G.) resistance in rice (Oryza sativa L.)	228
Seema Sinha, R Balasaraswathi*, K Selvaraju and P Shanmugasundaram
Methods for inhibition of residual lipase activity in colorimetric assay: A comparative study	233
Shamsher Singh Kanwar*, Rajeev Kumar Kaushal, Arshad Jawed, Reena Gupta and Swapandeep Singh Chimni
tRNomics: A comparative analysis of Picrophilus torridus with other archaeal thermoacidophiles	238
Bibekanand Mallick*, Jayprokas Chakrabarti, Smarajit Das and Zhumur Ghosh
Notes
Occurrence of trypsin-like protease in cardamom (Elettaria cardamomum  Maton)	243
A Josephrajkumar*, Romit Chakrabarty and George Thomas
Antioxidant activity of ethanolic extract of Terminalia chebula fruit against isoproterenol-induced oxidative stress in rats	246
Subramaniyan Suchalatha and Chennam Srinivasulu Shyamala Devi*
Biochemical changes in erythrocyte membrane in uncontrolled type 2 diabetes mellitus	250
Shamlan M S Reshamwala and Neela D Patil*
(Contd)
Effect of phenobarbitone on cytochrome P450 activity and chlorpyrifos and 3,5,6-trichloropyridinol levels in liver and serum in rat	254
Radhey S Verma, Anugya Mehta and N Srivastava*
Yolk protein profiles of three prawn (Macrobrachium) species during reproductive cycle	258
S Shanju and P Geraldine*
Book Review	262
Instructions to Authors	264

 

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*Author for correspondence

 

 

 

 

 

 

 

 

AUTHOR INDEX

 

 

 

 

 

 

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp. 205-215

 

Studies on surfactant-biopolymer interaction. II. Interaction of

cetyl trimethyl ammonium-, cetyl ethanolyl dimethyl ammonium-,

cetyl diethanolyl methyl ammonium- and cetyl triphenyl phosphonium

bromides and cetyl pyridinium chloride with calf thymus DNA

 

Arindam Chatterjee and Satya P Moulik*

 

The interaction of the surfactants cetyl trimethyl ammonium-, cetyl ethanolyl dimethyl ammonium-, cetyl diethanolyl methyl ammonium-, and cetyl triphenyl phosphonium bromides and cetyl pyridinium chloride with calf thymus DNA was studied at 303 K in phosphate buffer (pH 7.0) at 10 mM NaCl using spectrophotometric, viscometric, tensiometric, dynamic light scattering, circular dichroism, fluorescence microscopic and microcalorimetric techniques. All the surfactants interacted fairly with DNA, making the biopolymer condensed, even to the aggregated globular configuration at higher [surfactant]/[DNA] mole ratio (R), with direct evidence from fluorescence microscopy. Melting temperature and light scattering intensity of the DNA increased, whereas viscosity decreased in the presence of the surfactants. Tensiometry evidenced effective interaction at [surfactant] as low as 7.6 mM. Isothermal titration calorimetric measurements supported low enthalpy of binding, induced aggregation of the surfactants, increased critical micellar concentration and association of aggregates with the biopolymer at higher R, evidencing distinctions in thermal behaviour.

Keywords: Surfactants interaction, calf thymus DNA, spectrophotometry, viscometry, tensiometry, dynamic light scattering, circular dichroism, fluorescence microscopy, isothermal titration calorimetry

 

 

IPC Code: G01K3/00, G01N33/52

E -mail: cssju@yahoo.co.uk

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp. 216-221

 

 

Oxidative stress in tumour-bearing fore-stomach and distant normal organs of Swiss albino mice

Gagandeep¹, A R Rao and R K Kale*

 

Antioxidant status in the tumour-bearing fore-stomach and distant normal organs (liver, spleen, kidney and heart) was investigated in Swiss albino mice. In addition, the cytochrome P₄₅₀ (cyt P₄₅₀) system was also examined in the liver. Benzo(a)pyrene [B(a)P] (8 doses of 1 mg/0.1 ml) was administered twice a week for 4 weeks to develop fore-stomach tumour. The animals were sacrificed at the end of 140 days. The specific activities of catalase (CAT), DT-diaphorase (DTD) and glutathione-S-transferase (GST) were found decreased, and the level of reduced glutathione (GSH) and the specific activity of lactate dehydrogenase (LDH) increased in the tumour-bearing fore-stomach; however, no change was observed in superoxide dismutase (SOD) activity. The specific activities of antioxidant enzymes, and levels of GSH were also altered in the normal organs, depending upon the type of tissue. In addition, the contents of cyt P₄₅₀ and cyt b_5, and the activity of NADPH cyt P₄₅₀ reductase were significantly decreased in the liver. The results suggest increased oxidative stress in the tumour, and disturbance in the cooperative antioxidant functions in the distant normal organs. Inhibition of cyt P₄₅₀ system reflected the possible adverse effect on drug metabolism function of the liver. Since, the antioxidant potential and the drug metabolism function were altered, the findings may have relevance to the radiation and chemotherapy of cancer.

Keywords: fore-stomach tumour, oxidative stress, antioxidant enzymes, carcinogen metabolizing enzymes, benzo(a)pyrene, cytochrome P₄₅₀ system

 

IPC Code: C07M 3/00

          E-Mail: rkkale@hotmail.com

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp 222-227

 

 

Induction of apoptosis in pheochromocytoma (PC12) cells exposed to eicosapentaenoic acid in vitro

Mei Li, Haitao Ge, Xiuqin Kong, Weifa Zheng¹ and Zhili Liu*

 

The effect of different concentrations of eicosapentaenoic acid (EPA), on rat pheochromocytoma PC12 cells were evaluated using cell viability, lactate dehydrogenase (LDH) activity, flow cytometric DNA analysis and electronic microscopy. A time- and dose-dependent decrease in the cell viability was observed in the cultures of PC12 cells, supplemented with EPA. The incubation with 200 μM EPA for 48 and 72 hr induced a decrease in the cell viability by 53.40 and 53.43%, respectively. Treatment of PC12 cells with EPA induced apoptosis in a concentration-dependent manner, as evidenced by flow cytometry analysis. The highest percentage of apoptotic cells accumulated to 30.32%, following treatment with 200 μM EPA. The LDH levels increased significantly on treatment with 100 and 200 μM EPA, by 144.4 and 197.3%, respectively, compared with the untreated control. In addition, the cell morphology change was also observed by electron microscopy. The results suggest that EPA mediates its effect on the PC12 cells, at least in part, via the induction by apoptosis.

Keywords: Eicosapentaenoic acid, pheochromocytoma PC12, cell viability, lactate dehydrogenase, flow cytometric DNA analysis, electronic microscopy, apoptosis

 

IPC Code: G01 N 33/574, 901 J 37/26

          E-mail: liuzl@nju.edu.cn

 

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp. 228-232

 

 

Molecular and biochemical markers associated with leaffolder

(Cnaphalocrocis medinalis G.) resistance in rice (Oryza sativa L.)

 

Seema Sinha, R Balasaraswathi*, K Selvaraju and P Shanmugasundaram

 

Association of molecular markers namely isozymes and simple sequence repeats (SSRs) and various biochemical markers to leaffolder (Cnaphalocrocis medinalis G., a predominant insect pest of rice) resistance were studied in rice (Oryza sativa L.). Recombinant inbred lines (RILs) of F₈ generation obtained by crossing IR36 (susceptible parent) and TNAULFR831311 (moderately resistant parent) were used in this study. Soluble protein content, protein profile, and peroxidase and phenylalanine ammonia lyase (PAL) activities were the various biochemical markers studied. Decrease in soluble leaf protein content was observed in all lines, due to insect infestation. Protein profiling revealed an enhanced expression of a high molecular mass (>97 kDa) protein in all the infested lines. Besides, there was an increased induction of a 38 kDa protein in infested resistant parent and resistant RILs. A significant increase in peroxidase and PAL activities was observed after infestation. In peroxidase isozyme analysis, carried out after infestation, “isoform 1” was found to be more prominent in the susceptible lines and “isoform 2” in the resistant lines. Bulk segregant analysis (BSA) with twenty-five rice microsatellites (RM) resulted in identification of three polymorphic markers between bulks RM11 and RM432 located on chromosome 7 and RM271 on chromosome 10 of rice. These markers may be associated with leaffolder resistance in rice and can be used in marker-assisted selection for leaffolder resistance in rice.

Keywords: Rice, Oryza sativa L, simple sequence repeat, isozyme, leaffolder, Cnaphalocrocis medinalis G., peroxidase, phenylalanine ammonia lyase, SDS-PAGE, bulk segregant analysis, leaf protein profile, soluble protein.

 

 

IPC Code: C12N15/65, C12N15/52

          E-mail: rbsara @ rediffmail.com

 

 

 

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp. 233-237

 

 

Methods for inhibition of residual lipase activity in colorimetric assay:
A comparative study

 

Shamsher Singh Kanwar*, Rajeev Kumar Kaushal, Arshad Jawed, Reena Gupta and Swapandeep Singh Chimni¹

 

A comparative study of various treatments for inhibition of the residual activity of a lipase (obtained from Bacillus coagulans MTCC-6375) in a colorimetric assay using p-nitrophenyl palmitate (pNPP) was made. Direct chilling of contents of reaction mixture or addition of chilled mixture of ethanol : acetone (1:1) decreased the residual lipase activity by 94.0 and 95.0% respectively, as compared to lipase incubated at 45^oC for 20 min (control). Amongst various ionic and non-ionic detergents, Triton X-100 (0.07%, v/v) and sodium lauryl sarcosine or SLS (0.25%, w/v) partially, and SDS (0.05%, w/v) completely blocked the residual lipase activity of B. coagulans lipase in colorimetric assay. Addition of a serine protease inhibitor, PMSF (15 mM) or EDTA (200 mM) inhibited residual lipase activity by 99.5 and 100%, respectively. However, addition of reducing agents viz., 2-mercaptoethanol and dithiothreitol caused decomposition of chromogenic substrate (pNPP) thus rendering the colorimetric method unfit for lipase assay. EDTA (200 mM) and SDS (0.05%, w/v) were also highly effective in inhibiting the residual activities of lipases of Pseudomonas aeruginosa MTCC-4713, P. cepacia and commercial grade lipolytic preparations such as lipozyme, lipolase and porcine pancreatic lipase. However, PMSF (15 mM) completely inhibited the residual activity of lipase of P. aeruginosa.

Keywords: Bacillus coagulans MTCC-6375, microbial and commercial lipases, chilling, ethanol:acetone mixture, denaturing/reducing/chelating agents, PMSF.

IPC Code: C12N1/00; C12N9/00; G01N33/52

           E-mail: kanwarss2000@yahoo.com

 

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp. 238-242

 

 

 

tRNomics: A comparative analysis of Picrophilus torridus
with other archaeal thermoacidophiles

 

Bibekanand Mallick^*, Jayprokas Chakrabarti^†, Smarajit Das and Zhumur Ghosh

 

In the euryarchaeal thermoacidophile Picrophilus torridus DSM 9790, we identified a copy of rare tRNA^Ile(TAT) gene, along with the other 47 tDNAs with the help of our in-house program. Further, tRNAs of P. torridus were also compared with other archaeal thermoacidophiles Thermoplasma acidophilum, T. volcanium, Sulfolobus solfataricus and S. tokodaii.

          Keywords: tRNomics, Picrophilus torridus, overlapping tRNA, base pairs, intron, Archaea, thermoacidophile

          E-mail: tpbm@iacs.res.in; vivekm@iitian.iitkgp.ernet.in

 

 

 

 

 

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp. 243-245

 

 

 

 

Occurrence of trypsin-like protease in cardamom (Elettaria cardamomum  Maton)

A Josephrajkumar*, Romit Chakrabarty^$ and George Thomas^#

 

Occurrence of trypsin-like protease in fresh cardamom (Elettaria cardamomum  Maton) seeds, as evidenced by the benzoyl-arg-p-nitroanilide (BApNA) hydrolyzing ability of the seed enzyme preparation under alkaline condition is reported for the first time. The enzyme has a pH and temperature optima as 8 and 45^oC, respectively. It is inhibited by aprotinin and phenylmethyl sulfonyl fluoride (PMSF) in a dose-dependent manner, suggesting the presence of serine residues at the active site. The enzyme had a V_max of 98.01 nmoles p-nitroaniline released per min per mg protein and K_m of 0.0684 mM with BApNA as substrate. Addition of aprotinin (75.75 mM) increased K_m value by three-folds, whereas the V_max was reduced by 23%.

 

Keywords: Cardamom, Elettaria cardamomum  Maton, trypsin, aprotinin.

 

IPC Code: A61K38/43,  C12N9/00

E-mail: entojoe2003@yahoo.co.in

 

 

 

 

 

 

 

 

 

 

 

Indian Journal of Biochemisty & Biophysics

Vol. 42, August 2005, pp. 246-249

 

 

 

Antioxidant activity of ethanolic extract of Terminalia chebula fruit against isoproterenol-induced oxidative stress in rats

Subramaniyan Suchalatha and Chennam Srinivasulu Shyamala Devi^*

 

Antioxidant activity of ethanolic extract of fruits of Terminalia chebula (500 mg/kg body wt, orally for 30 days) against isoproterenol-induced oxidative stress was investigated in rats. The levels of serum lipid peroxides, iron, ascorbic acid, vitamin E, plasma iron-binding capacity, and the activities of ceruloplasmin and glutathione were assayed, in addition to the activities of the antioxidant enzymes — glutathione peroxidase, glutathione reductase, glutathione-S-transferase, superoxide dismutase and catalase in the heart tissue. Administration of isoproterenol increased the levels of lipid peroxides and iron, with corresponding decrease in the activities of the enzymic and non-enzymic antioxidants. The pre-treatment with ethanolic extract of fruits significantly prevented the alterations induced by isoproterenol, and maintained a near normal antioxidant status. Results suggest that the cardioprotective effect of T. chebula fruit may partly be attributed to its antioxidant properties.

          Keywords: Terminalia chebula, isoproterenol, myocardial infarction, lipid peroxidation, antioxidant.

          IPC Code: A61P9/00, A61P9/10

          E-mail: cssdevi@yahoo.com

 

 

 

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp. 250-253

 

 

Biochemical changes in erythrocytemembrane in uncontrolled type
2 diabetes mellitus

 

Shamlan M S Reshamwala and Neela D Patil*

 

The levels of lipid peroxidation and alterations in lipid composition and ATPase activities were determined in erythrocyte plasma membrane of uncontrolled type 2 diabetes mellitus (DM) patients. The study groups consisted of 30 patients (16 males, 14 females) attending the Out Patients’ Department of Lokmanya Tilak Municipal General Hospital, Mumbai, and 23 age- and sex-matched control subjects (15 males, 8 females). Glycated haemoglobin (an index of long-term glycaemic control), erythrocyte membrane cholesterol, phospholipid and cholesterol to phospholipid molar ratio, lipid peroxidation products in the form of thiobarbituric acid-reactive substances (TBARS), and Na⁺-K⁺-ATPase activity were found to be significantly increased, and Mg²⁺-ATPase activity significantly decreased, in the diabetic subjects, as compared to controls. The study suggests that biochemical changes in the erythrocyte membrane may be involved in the pathophysiology of type 2 DM.

Keywords:            Uncontrolled type 2 diabetes mellitus, erythrocyte membrane, glycated haemoglobin, Na⁺-K⁺-ATPase, Mg²⁺-ATPase, lipid peroxidation, cholesterol, phospholipid

IPC Code: A61K38/42; A61P7/00

E-mail: neelap29@yahoo.com

 

 

 

 

 

 

 

Indian Journal of Biochemisty & Biophysics

Vol. 42, August 2005, pp. 254-257

 

 

 

Effect of phenobarbitone on cytochrome P450 activity and chlorpyrifos and 3,5,6-trichloropyridinol levels in liver and serum in rat

Radhey S Verma, Anugya Mehta and N Srivastava*

 

 

Chlorpyrifos [O,O¢-diethyl-O-(3,5,6-trichloro-2-pyridyl) phos-phorothionate, CPF] undergoes oxidative desulfuration or dearylation by hepatic microsomal cytochrome P450 (CYP)-mediated monooxygenase reaction to CPF oxon or desethyl CPF, which are further metabolized to 3,5,6-trichloropyridinol (TCP). The present study showed that CPF exposure caused induction of hepatic CYP levels in rats. Phenobarbitone (PB) treatment elevated CYP activity by about 2.2-folds in controls, while CPF exposure to PB-treated rats did not cause further elevation in CYP levels. The levels of CPF in liver tissue and serum of rats given 50 and 100 mg CPF/kg body wt for 3 weeks were 17.15 ng and 29.39 ng/g and 33.71 ng and 56.34 ng/ml, respectively. The levels of TCP in these rats were 123.58 ng and 215.26 ng/g in liver tissue and 391.73 ng and 599.32 ng/ml in serum respectively. On PB-treatment, CPF levels were decreased by 46% and 38% in liver and 23% and 20% in serum of rats receiving 50 mg and 100 mg CPF/kg body wt for 3 days, while TCP levels were increased by 6% and 22% in liver and 22% and 44% serum, respectively. Results of this study clearly show that CYP2B, the PB-inducible form can biotransform CPF faster into TCP.

 

Keywords: Organophosphate pesticides, chlorpyrifos, cytochrome P450, 3,5,6-trichloropyridinol, pheno-barbitone, hepatic microsome, cytochrome P450 induction, serum, liver, rat

 

IPC Code: A01N55/00

E-Mail: nalinis21@yahoo.com

 

 

 

 

 

 

Indian Journal of Biochemistry & Biophysics

Vol. 42, August 2005, pp. 258-261

 

 

Yolk protein profiles of three prawn (Macrobrachium) species during reproductive cycle

 

S Shanju and P Geraldine*

 

Ovarian yolk protein (vitellin) of adult freshwater prawns Macrobrachium malcolmsonii, M. rosenbergii and M. lamarreii was characterized, and relationship between the gonadosomatic index (GSI) and the appearance of specific polypeptides in their ovary during different stages (early-mature, mature and spent) of reproductive cycle was studied. Mature ovaries of all the three species showed increased GSI values, compared to the respective early-mature ovaries. Protein profiles at the three stages of ovarian maturation were analysed by SDS-PAGE. Polypeptides of low molecular mass (20 to 70 kDa) were prominent in the early-mature ovary, whereas in mature ovary, 89 and 100 kDa polypeptides were predominant. Immunodiffusion studies, using antiserum raised against purified vitellin from mature ovary of
M. malcolmsonii indicated antigenic similarities of vitellins among the three species.

Keywords:   Yolk protein, ovary, Macrobrachium species, vitellin, gonadosomatic index, immunodiffusion

IPC Code: C07K1/26; C25B7/00

E-mail: Geraldine_Philip@yahoo. co.in

 

 

 

 

 

 

 

Indian Journal of Biochemisty & Biophysics

Vol. 42, August 2005, pp. 262-263

 

 

 

 

BOOK REVIEW

 

Chlorophyll a Fluorescence: A Signature of Photosynthesis: Edited by: George C Papageorgiou and Govindjee, Published by: Springer, Dordrecht, The Netherlands, 2004; ISBN: 1-4020-32170XHB; Hard Copy: Bound, Page - 818; Price: Euro 240
(Rs. 14, 000 Approx.)

     The tiny little red light emitted from green pigments of plants, algae and cyanobacteria provides huge information on the vital plant processes – from light capture and utilization of quanta in photochemistry to its down-stream processing in carbon and nitrogen assimilation and regulation. Chlorophyll (Chl) fluorescence has had wide applications in plant biology research in recent years, and is used by photosynthesis researchers, crop scientists, ecologists and town planners. Thus, are a large number of books, reviews and circulars appearing regularly on Chl fluorescence from photosynthetic systems. The current volume 19 of AIPH series ‘Chlorophyll a fluorescence: A signature of photosynthesis’ edited by Prof. George C Papageorgiou of NRC Demokritos, Greece and Prof. Govindjee of University of Illinois, Urbana, Illinois, USA, two pioneers in this field, is one of the most comprehensive treatises on this subject. The two editors successfully persuaded 54 experts to contribute 31 authoritative chapters on both basic and applied aspects of Chl a fluorescence by photosynthetic organisms.

Starting with a bit of basic and history of Chl a fluorescence and on the fluorescence characteristics of photosynthesis pigments in vitro and in vivo, the volume expands into many basic and applied features of Chl a fluorescence. Among the basic aspects, the topics like excitation energy migration and energy transfer, trapping of excitation energy in photosystems, and trapping models of photosystem (PS) II include new developments in the area. Also, specific topics such as photon capture by cyanobacteria, PS II Chl a fluorescence yield changes under multiple flash-regime as well as the fluorescence of photosystems in the red algae, discussed in this book, are of interest to photosynthetic researchers.

The development of Pulse Amplitude Modulated (PAM) fluorometry has made the Chl a fluorescence ‘global’. Analysis of fluorescence fast-transient parameters has provided new dimensions for studies on photophysiological processes in plants. The developments of guidelines on the measurements of Chl a fluorescence, as well as new user-friendly equipment have made Chl a fluorescence studies a very popular technique in plant research. All these aspects have been vividly described in this volume.

The volume also describes the use of Chl fluorescence in probing photosynthesis performance and plant productivity, in remote sensing as well as metal toxicity and other environmental stresses. Chl fluorescence is used in transgenic plants as well as state transition mutants. Besides, the book presents the valuable development in Chl a fluorescence imaging that has added new dimension to the monitoring power of this technique. Imaging is not only used in indexing the photosynthetic performance of crop plants but also in estimating the exact extent of biotic and abiotic stresses and associated damages to the crop. Chl a fluorescence transition has been successfully used in monitoring primary processes in aquatic, marine and wetland ecosystems, and these topics have been adequately discussed in the book. Chl a fluorescence signatures, can be studied and measured in a variety of oxygenic photosynthetic organisms as well as at numerous biological diversities.

The major importance of the present volume lies in providing the readers the wide spectrum of use of Chl a fluorescence technique and providing considerable guidelines for its numerous applications. The book is, therefore, likely to satisfy both the professional and general readers. It can be used as a reference text for courses in photosynthesis, crop sciences, and stress physiology. Like the previous volumes of the series, the present volume is printed on high quality paper and contains illustrations (graphs, charts and photographs). Although the book is bulky in size and cost, but that should not deter anyone to purchase this volume. The book is a worth addition to the collection for students, faculty and libraries as well.

We hope that the readers would appreciate the efforts that the two editors and international authors (including Prof. Prasanna Mohanty and Dr Manoj Joshi from India) have put in bringing out such a quality publication. The editors and all the contributors deserve appreciation from all of us.

B Vani

Biosciences and Biotechnology, BITS, Pilani, India

Sujata R Mishra

Dept of Molecular Biology, Pusan National University, South Korea

G B Kashpuri

LBG House, Bhubaneswar, India

E mail: photosis@rediffmail.com