018460   Mukhopadhyay R;Bishayi B (Immunology Lab, Physiology Dep, University of Calcutta, University Colleges of Science and Technology, 92 A.P.C. Road, Kolkata-700 009, Email: biswa_dev2@yahoo.com) : Effects of soluble antigen-induced immune cell activation on steroidogenesis in murine lymphoid organ. Indian J Biochem Biophys 2008, 45(4), 250-5.

The effect of soluble antigenic (bovine serum albumin, BSA) stimulation to induce steroidogenesis in murine lymphoid organs with concomitant changes in proinflammatory or inflammatory cytokine levels and its implication in the alteration of T-cell response was studied in the mice. Male Swiss albino mice (6-8 weeks old) with average body weight (20±4 g) were randomly assigned to 3 groups and injected with BSA in presence and absence of Freund's complete or incomplete adjuvant, whereas the control group received only saline. After 3 weeks, animals were sacrificed, and serums as well as lymphoid organs were collected. From the lymphoid tissue homogenate, the activities of steroidogenic enzymes and corticosterone and cytokine levels of the serum were estimated. Steroidogenic enzyme activities in murine lymphoid organs, as well as the pro-inflammatory and inflammatory cytokines levels in serum increased after Freund's complete adjuvant-emulsified BSA administration, as compared to control. The serum corticosterone and serum cytokine profile were also elevated. Results suggested that soluble protein antigen (BSA) administration stimulated steroidogenesis in murine lymphoid tissues and rise in the pro-inflammatory or inflammatory cytokine levels might indicate monocyte recruitment as well as TH₁ activation.

018459   Mohanty G;Mukherji S (Centre for Environmental Science and Engineering (CESE), Indian Institute of Technology Bombay, Powai, Mumbai-400 076) : Enhancement of NAPL bioavailability by induction of cell-surface hydrophobicity in Exiguobacterium aurantiacum and Burkholderia cepacia. Indian J Biotechnol 2008, 7(3), 295-306.

Induction of cell surface hydrophobicity in bacterial cultures can facilitate the direct interfacial uptake of non-aqueous phase liquids (NAPLs). This study explores bioavailability of NAPLs for Exiguobacterium aurantiacum and Burkholderia cepacia isolated from oil-contaminated soil and sediments. Surface tension measurements and emulsification activity tests did not provide evidence for release of extracellular biosurfactants/bioemulsifiers. Contact angle measurement on cell layers and bacterial adhesion to hydrocarbon (BATH) assay was conducted for determining the cell surface hydrophobicity. While the surfaces of cultures grown on soluble substrate, dextrose, were not hydrophobic, higher water contact angle and greater adherence to n-hexadecane/diesel revealed the highly hydrophobic nature of the cell surfaces for cultures grown on NAPLs (n-hexadecane and diesel), thus providing evidence for induction in cell surface hydrophobicity. Positive results in the carbocyanine assay indicating release of lipopolysaccharides/extracellular polysaccharides was observed over the log growth phase only for the NAPL-grown cells. Transmission electron microscopy (TEM) revealed an abundance of intracellular electron transparent globules within the NAPL-grown cells. Light microscopy and TEM images together revealed differences in cell surface characteristics of E. aurantiacum and B. cepacia, which was also affected by the growth substrate.

018458   Mishra N;Gupta P N;Khatri K;Goyal A K;Vyas S P (Drug Delivery Research Lab, Pharmaceutical Sciences Dep, Dr Harisingh Gour Vishwavidyalaya, Sagar 470 003) : Edible vaccines: a new approach to oral immunization. Indian J Biotechnol 2008, 7(3), 283-94.

Edible vaccines offer exciting possibilities for significantly reducing the burden of diseases like hepatitis and diarrhoea, particularly in developing world where storing and administering vaccines are the major problems. Edible vaccines are prepared by molecular farming using the science of genetic engineering. Selected genes are introduced into the plants. The transgenic plant is then induced to manufacture the encoded protein. Owing to its low cost, it will be suitable for developing countries like India. Edible vaccines are mucosal-targeted vaccines, which cause stimulation of both systematic and mucosal immune response. Edible vaccines are being developed for various diseases, such as measles, cholera and hepatitis B, and many more are in the process of development. Thus, they may also help to suppress autoimmune disorders such as Type-I diabetes, diarrhoea, multiple sclerosis, rheumatoid arthritis, etc. Human trials conducted by the National Institute of Allergy and Infectious Diseases (NIAID), US Department of Health and Human Services, USA show that edible vaccines are feasible. ProdiGene, a biotech company, has a patent for vaccine against, viral diseases of hepatitis and transmissible gastroenteritis virus. This review comprises methods of preparation, mechanism of action, recent developments, clinical trials and therapeutic applications of edible vaccines.

018457   Minakshi;Pundir C S (Biochemistry Research Lab, Biochemistry & Genetics Dep, MD University, Rohtak-124 001, Email: pundircs@rediffmail.com) : Co-immobilization of lipase, glycerol kinase, glycerol-3-phosphate oxidase and peroxidase on to aryl amine glass beads affixed on plastic strip for determination of triglycerides in serum. Indian J Biochem Biophys 2008, 45(2), 111-5.

Commercial lipase, glycerol kinase (GK), glycerol-3-phosphate oxidase (GPO) and peroxidase (POD) have been co-immobilized covalently on to arylamine glass beads affixed on a plastic strip through diazotization with a conjugation yield of 89.1 mg/g support and 64.1% retention of specific activity. The co-immobilized enzymes showed maximum activity at pH 7.5, when incubated at 40°C for 20 min. The strip was employed for determination of serum triglycerides (Tgs). The minimum detection limit of the method was 0.20 mM/L. The recovery of added Tgs was 88.0%. Within day and between day coefficient of variations were <60>7.0% and <60>11.0%, respectively. A good correlation (r = 0.982) was observed between total serum Tgs values obtained by present method and the most commonly used enzymic colorimetric method, employing free enzymes. Among the various serum substances tested at their physiological concentrations, only cholesterol, ascorbic acid and bilirubin caused 30%, 15%, and 20% inhibition of strip-bound enzymes, respectively. The strip lost 50% of its activity after 150 regular uses over a period of 33 days, when stored in reaction buffer at 4°C. The method reported here has the advantage over other existing methods, as it provides higher sensitivity, better stability and reusability of co-immobilized enzymes and is also economical.

018456   Manohari C;Backiyarani S;Jebasingh T;Somanath A;Usha R (Plant Biotechnology Dep, School of Biotechnology, Madurai Kamaraj University, Madurai- 625 021) : Efficient plant regeneration in small cardamom (Elettaria cardamomum Maton.) through somatic embryogenesis. Indian J Biotechnol 2008, 7(3), 407-9.

Efficient protocol for the induction of somatic embryogenesis and plant regeneration in small cardamom (Elettaria cardamomum Maton.) from the inner core region of rhizome has been established. Calli developed profusely on Murashige and Skoog (MS) medium containing 9.0 μM 2,4 D and 2.3 μM Kn. When the friable calli were cultured in MS medium containing 4.4 or 8.8 μM BAP + 0.5 μM NAA, abundant embryogenic calli were obtained. The highest frequency of embryogenic calli (68%) and plantlets (86%) were obtained from MS medium containing 4.4 μM BAP and 0.5 μM NAA. Further shoot development was observed with 13.2 μM BAP + 0.5 μM NAA. In the same medium, roots also appeared, thereby eliminating an additional step of in vitro rooting. The well-developed plants were hardened and transferred to a mist chamber in a greenhouse with 90% survival frequency.

018455   Manikandan K;Saravanan V;Viruthagiri T (Chemical Engineering Dep, Annamalai University, Annamalai Nagar-608 002) : Kinetics studies on ethanol production from banana peel waste using mutant strain of Saccharomyces cerevisiae. Indian J Biotechnol 2008, 7(1), 83-8.

Five different mutant strains were developed from the wild strain of Saccharomyces cerevisiae (MTCC No.287) using UV irradiation technique by varying the exposure timings. All the mutant cultures were used for ethanol production using banana peel as a substrate in a batch fermenter. The effect of temperature, pH and initial substrate concentration on ethanol production were studied and optimized. The mutant strain 4 gave a maximum ethanol production of 9 g/L under identical conditions. The conditions were optimized for mutant strain 4 and a temperature of 33°C, pH 4.5 and initial substrate concentration 10%(w/v) were found to be optimum. The kinetics of ethanol production using mutant strain 4 under optimum conditions was studied and modeling was attempted using different models. Monod model for growth kinetics and Leudeking-Piret model for product formation kinetics were found to represent the experimental data very well.

018454   Mandal V;Sen S K;Mandal N C (P.G. Department of Botany, Darjeeling Govt. College, Darjeeling-734 101, Email: mandal_vivek@yahoo.co.in) : Optimized culture conditions for bacteriocin production by Pediococcus acidilactici LAB 5 and its characterization. Indian J Biochem Biophys 2008, 45(2), 106-10.

A strain of Pediococcus acidilactici LAB 5 was isolated from vacuum-packed fermented meat product, in order to obtain a novel bacteriocin from food-grade organisms. Optimized culture conditions for bacteriocin production in different media (viz., MRS, TGE, TGE + buffer, TGE + Tween 80, and TGE + Tween 80 + buffer) and at different temperatures and pH conditions were reported. TGE + Tween 80 + buffer medium was found to be most effective for bacteriocin production (about 2,400 AU/ ml) by this strain, when incubated at 37°C for 24 h. Bacteriocin, partially purified by adsorption-desorption method showed molecular mass of 10.3 kDa and produced prominent inhibition zone in activity gel. It showed significant storage stability both at high as well as in low temperatures for up to 6 months and retained its activity in a number of organic solvents, except in 2-mercaptoethanol. The treatment with amylase and lysozyme did not change its activity, but it lost its activity on proteinase K treatment. Antibacterial efficacy of bacteriocin was proved against some food spoilage and human pathogenic bacteria like Enterococcus, Leuconostoc, Listeria, Staphylococcus and Streptococcus.

018453   Liang C W;Zhanga X W;Tiana L;Oina S (NO, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, P.R. China, Email: sqin@ms.qdio.ac.cn) : Functional characterization of sll0659 from Synechocystis sp. PCC 6803. Indian J Biochem Biophys 2008, 45(4), 275-7.

Synechocystis sp. PCC 6803 lacks a gene for the any known types of lycopene cyclase. Recently, we reported that Sll0659 (unknown for its function) from Synechocystis sp. PCC6803 shows similarity in sequence to a lycopene cyclase gene-CruA from Chlorobium tepidum. To test, whether sll0659 encoded protein serves as lycopene cyclase, in this study, we investigated the carotenoids of the wild types and mutants. In the sll0659 deleted mutant, there is no blockage at the lycopene cyclization step. Our results demonstrate that sll0659 does not affect lycopene cycilzation. However, the ultrastructure of mutants suggests the involvement or necessity of sll0659 in the cell division.

018452   Leskovac V;Trivic S;Pericin D;Popovic M; Kandrac J (Faculty of Technology, University of Novi Sad, Bulevar Cara Lazara 1, 21000 Novi Sad, Serbia, Email: jkandrac@polj.ns.ac.yu) : Thermodynamic properties of the Calvin cycle and pentose phosphate pathway. Indian J Biochem Biophys 2008, 45(3), 157-65.

The enzymes of the Calvin cycle and pentose phosphate pathway operate in close conjunction with enzymes of glycolysis and gluconeogenesis. The last two metabolic routes are thermodynamically well characterized, but the former two are not. In this work, the thermodynamic properties of the 19 enzymatic reactions of the pentose phosphate pathway and the Calvin cycle, under standard conditions were calculated in the form of standard transformed reaction Gibbs energies at pH 5, 6, 7, 8, and 9, 25°C and ionic strength 0.25 M.

018451   Koti J S;Kanugula S;Suryanarayana T (Biochemistry Dep, School of Life Sciences, University of Hyderabad, Hyderabad-500 046, Email: tssl@uohyd.ernet.in) : DNA aggregation by an archaeal DNA binding protein Sac10b and its novel DNA nicking activity. Indian J Biochem Biophys 2008, 45(3), 166-73.

The solution structure of an archaeal DNA binding protein Sac10b (DBNP-B) by cross-linking with formaldehyde and its interaction with DNA were studied. Results indicated that Sac10b existed as oligomeric structure in solution and the oligomerization was greatly stimulated by Mg²⁺ ions and elevated temperatures. In light of our earlier observation that Sac10b interacts with DNA forming different types of complexes with DNA at different protein concentrations, its DNA-binding properties were also studied. Results demonstrated that the protein formed rapidly sedimentable co-aggregation complexes with both native and denatured DNA in a protein concentration-dependent manner. These protein DNA complexes were fluid-like crystalline material at a protein DNA ratio (3-6:1 w/w). Gel mobility shift assays carried out to study the interaction of the protein with plasmid DNA indicated possible DNA nicking by the protein. The DNA nicking activity of Sac10b was optimal in pH range of 7-8.5 and was dependent on Mg²⁺ ions. It was maximal at protein to DNA ratio of (8:1, w/w) and very little activity was observed above and below this ratio. Nicking of DNA at this ratio indicated structure-specific DNA nicking by the protein. The protein might have important multi-functional role in the DNA metabolism in this organism.

018450   Kizhakkayil J;Thomas E;Sasikumar B (Crop Improvement and Biotechnology Div, Indian Institute of Spices Research, Marikunnu P O, Calicut-673 012) : Molecular characterization of traded black pepper (Piper nigrum L.) from India, Indonesia, Vietnam and Malaysia. Indian J Biotechnol 2008, 7(2), 210-3.

Molecular profiling (RAPD) and clustering of traded black pepper samples from India, Indonesia, Vietnam and Malaysia revealed a comparatively high genetic similarity within the samples from a particular country than between any two countries. The UPGMA dendrogram constructed based on the similarity coefficient revealed a total of four groups in two different clusters. The two Indonesian samples form cluster I, while others form cluster II. The genuine Indian varieties and the traded pepper from India formed separate group in cluster II. Similarly, black pepper from Malaysia and Vietnam also formed distinct groups in cluster II. The aspect of genetic similarity was discussed in relation to the origin and spread of black pepper.

018449   Kiranmai C;Aruna V;Karuppusamy S;Pullaiah T (Dep of Biotechnology and Botany, Sri Krishnadevaraya University, Anantapur-515 003, Email: pullaiaht@yahoo.co.in) : In vitro propagation of Pergularia daemia (Forsk.) chiov. Pl Cell Biotechnol molec Biol 2007, 8(1-2), 67-72.

Study was conducted to establish a protocol for rapid propagation of Pergularia daemia (Asclepiadaceae) through different aseptic seedling explants. 40 days old aseptic seedling grown on half stength MS medium were used as explant. All the explants were cultured on MS medium containing different concentration of cytokinins alone. MS medium supplemented with BAP 8.87μM produced maximum number of 11.20 shoots with the shoot tip explant. Nodal explant on BAP 13.3μM attained maximum shoot length of 4.78cm. High frequency of rooting (85%) and maximum number of 6.2 roots were produced on half strength MS medium supplemented with IBA 0.49μM. After acclimatization, 70% of the micropropagated plantlets were successfully esablished in the field.

1 illus, 2 tables, 14 ref

018448   Kaur J;Kaul G (Animal Biochemistry Dep, National Dairy Research Institute, Karnal, Haryana-132 001, Email: gkndri@gmail.com) : Protein translocation pathways across the inner and outer mitochondrial membranes. Indian J Biochem Biophys 2008, 45(3), 149-56.

Mitochondria import different proteins that are encoded by nuclear genes and synthesized in the cytosol. Separate translocases in inner and outer mitochondrial membranes like TOM, TIM23, TOB/SAM, TIM22 complex facilitate recognition, import and intramitochondrial sorting of preproteins. Various cytosolic factors as Hsp70 and auxillary factors assist in targeting these preproteins to their destinations. Also, different protein components in the matrix participate in this energetically driven translocation process in a reaction that depends upon membrane potential and matrix-ATP. This review summarizes the present knowledge on import and sorting of mitochondrial precursor proteins with glance on unresolved questions.

018447   Kathiresan K;Thiruneelakandan G (Centre of Advanced Study in Marine Biology, Annamalai University, Parangipettai-608 502) : Prospects of lactic acid bacteria of marine origin. Indian J Biotechnol 2008, 7(2), 170-7.

Lactic acid bacteria (LAB) are beneficial bacteria. They are known for their human gut floral maintenance, fermentative activity and food preservative capacity. The marine strains may have better potential than their terrestrial counterparts. This is due to essential nutrients provided by marine biotopes for nurturing the LAB, and to extreme environmental niches. The marine LAB and their by-products may have potential values in food processing, fermentation, pharmaceutical and biopolymer industries.

018446   Karthickeyan S M K;Kumarasamy P;Sivaselvam S N;Saravanan R;Thangaraju P (Animal Genetics and Breeding Dep, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences, Chennai-600 007) : Analysis of microsatellite markers in Ongole breed of cattle. Indian J Biotechnol 2008, 7(1), 113-6.

Evaluation of genetic variability was carried out in Ongole cattle using 25 microsatellite markers, recommended by FAO. All the screened loci were polymorphic and a total of 97 alleles were observed across the analyzed loci. The mean number of alleles was found to be 3.88±0.25 with a range of 2 to 7. The allele size ranged from 94 to 300 bp. The frequency distribution of microsatellite alleles in the breed was from 0.0119 to 0.9375. The estimated expected heterozygosity value was 0.6079±0.04 and the PIC was 0.5584±0.04. Microsatellite analysis revealed high allele and gene diversity in the investigated cattle and overall mean FIS value (-0.0615) suggested the excess of heterozygosity in the population. In addition, the high information of the polymorphic loci throw light on the scope for maintaining variation in the population and strategies to formulate conservation and further improvement measures.

018445   Jose S;Girija D;Beena P S (Centre for Plant Biotechnology and Molecular Biology, College of Horti, Kerala Agricultural University, Thrissur-680 656) : Cloning and characterization of the gene encoding HMG CoA reductase from black night shade (Solanum nigrum L.). Indian J Biotechnol 2008, 7(1), 66-72.

The first committed step in the pathway for biosynthesis of isoprenoids in plants is catalyzed by 3-hydroxy 3- methylglutaryl CoA reductase (HMGR; EC:1.1.1.34). Here we report for the first time, the cloning of a partial genomic DNA sequence encoding HMGR from a medicinal herb, Solanum nigrum L. This plant is associated with resistance against potato cyst nematode and the late blight pathogen, Phytophthora infestans. The clone Snhmgr (GenBank Acc.No. DQ 229901) had 582 base pairs (bp) with a 462 bp open reading frame (ORF), encoding 142 amino acid polypeptide. The in-silico analysis of Snhmgr sequence revealed about 85% identity with hmgr genes in other solanaceous plants. Phylogenetic analysis indicated that Snhmgr was more identical with tomato hmgr on evolutionary basis and belonged to the solanaceous cluster, including hmgr genes from tomato, potato, tobacco and capsicum. Functional analysis of conserved domains of Snhmgr showed CoA reductase, NAD binding and substrate binding activities.

018444   Jebasingh T;Jacob T;Shah M;Das D;Krishnaswamy S;Rsha R (School of Biotechnology, Madurai Kamaraj University, Madurai-625 021, Email: abayamba@gmail.com) : Optimized expression, solubilization and purification of nuclear inclusion protein b of Cardamom mosaic virus. Indian J Biochem Biophys 2008, 45(2), 98-105.

All RNA viruses encode an RNA-dependent RNA polymerase (RdRP) that is required for replication of the viral genome. Nuclear inclusion b (NIb) gene codes for the RdRp in Potyviridae viruses. In this study, expression, solubilization and purification of NIb protein of Cardamom mosaic virus (CdMV) is reported. The objective of the present study was to express and purify the NIb protein of CdMV on a large scale for structural characterization, as the structure of the RdRp from a plant virus is yet to be determined. However, the expression of NIb protein with hexa-histidine tag in Escherichia coli led to insoluble aggregates. Out of all the approaches [making truncated versions to reduce the size of protein; replacing an amino acid residue likely to be involved in hydrophobic intermolecular interactions with a hydrophilic one; expressing the protein along with chaperones; expression in Origami cells for proper disulphide bond formation, in E. coli as a fusion with maltose-binding protein (MBP) and in Nicotiana tabacum] to obtain the RdRp in a soluble form, only expression in E. coli as a fusion with MBP and its expression in N. tabacum were successful. The NIb expressed in plant or as a fusion with MBP in E. coli can be scaled up for further work.

018443   Jayashree R;Rekha K;Kumari Jayasree P;Sanju R;Thulaseedharan A (Biotechnology Div, Rubber Research Institute of India, Kottayam-686 009, Email: radha_jayasree@rediffmail.com) : Effect of colchicine on callus induction from isolated microspores in Hevea brasiliensis. Pl Cell Biotechnol molec Biol 2007, 8(1-2), 89-92.

Method for the induction of haploid callus from isolated microspores of Hevea brasiliensis has been developed. The growth promoting effects of colchicine on callus formation was proved. Anthers at the late uni-nucleate to the early bi-nucleate stage of the microspores were pretreated at 4°C for 5-7 days before microspore isolation. Different durations of colchicine exposure (24-96 hrs) as well as varying concentrations (100-1000 mg/l) were tested after the pre-culture. The maximum response was noticed when the microspores were pre-cultured in 100 mg/l colchicine for 96 hrs. It was observed that a high sucrose concentration (180 g/l) favored callus growth in the presence of the growth regulators, kinetin and zeatin (3.0 mg/l). This is a major step towards utilization of haploid technology in Hevea breeding.

2 tables, 23 ref

018442   Jayaram K;Prasad M N V (School of Life Sciences, Plant Sciences Dep, University of Hyderabad, Hyderabad-500 046) : Rapid in vitro multiplication of Drosera burmanii Vahl.: a vulnerable and medicinally important insectivorous plant. Indian J Biotechnol 2008, 7(2), 260-5.

Protocol has been stardardised for the rapid and large-scale in vitro multiplication of the vulnerable medicinal herb, Drosera burmanii Vahl. by enhanced axillary bud proliferation from shoot tip explants. In order to standardize in vitro multiplication, the effects of different strengths of Murashige and Skoog (MS) medium (1/4, 1/3, 1/2, and full strength), different percentages of sucrose (1%, 2% and 3%), various pH (3.7, 4.7, 5.7 and 6.7) and MS basal medium fortified with different concentrations of kinetin (Kn) and 6-benzylaminopurine (BAP) (0.1, 0.5, 1.0 and 2.0 mg L^-1 ) were tried on shoot tip explants. Maximum number of multiple shoots developed on MS medium supplemented with Kn (1.0 and 2.0 mg L^-1) and BAP (0.5, 1.0 and 2.0 mg L^-1) separately. Direct plantlet regeneration from the leaves and in vitro flowering were also observed. Rooting was best achieved on MS basal medium. This protocol could be useful for large-scale production of biomass for quercetin, plumbagin bioprospection and long term in vitro conservation.

018441   Jaswal R K;Kocher G S;Virk M S (Microbiology Dep, Punjab Agricultural University, Ludhiana-141 004) : Production of alkaline protease by Bacillus circulans using agricultural residues: a statistical approach. Indian J Biotechnol 2008, 7(3), 356-60.

Bacillus circulans, an alkaline protease producer isolated from vegetable waste, showed a pH of 10.5, temperature between 25-30°C and agitation rate of 200 rpm as optimum physical conditions for enzyme production. Factorial experiments were designed and the best CN combination of glucose and soybean meal produced a maximum alkaline protease of 461.65 U/mL. Soybean meal replaced with cotton deoiled meal (CDM) from enzyme production medium as the maximum supporter of alkaline protease produced 589 U/mL of protease. Protease values with different concentrations of CDM fitted in a regression equation showed that B. circulans produces maximum alkaline protease of 808.68 U mL^-1 in 100.9 h using 0.5789% CDM.

018440   Ilaiyaraja N;Rajarani A P;Santha I M (Biochemistry Div, Indian Agricultural Research Institute, New Delhi-110 012, Email: ims_bio@yahoo.com) : Cloning and characterization of diacylglycerol acyltransferase (DGAT) cDNA sequence from Brassica juncea cv. Pusa Bold. Indian J Biochem Biophys 2008, 45(1), 30-6.

Diacylgycerol acyltransferase (DGAT: EC 2.3.1.20) is the only enzyme in the Kennedy pathway that is exclusively committed to the synthesis of storage oil in plants. In this study, cloning and characterization of DGAT gene sequence from Brassica juncea cv Pusa bold, an important oil seed crop of India is reported. A partial gene sequence of 2003 bp was PCR amplified and cloned from B. juncea. Sequence analysis showed that it has 10 exonic and 9 intronic sequences in the partial gene. Two cDNA sequences namely BjDGAT 1 and BjDGAT 2 (1.5 kb) encoding DGAT enzymes were amplified by RT-PCR from the developing seeds. The complete length of these two cDNAs as determined by RACE technique was 1768 bp, including 5' and 3'-UTR. Comparative analysis of the sequences showed that BjDGAT1 was 85.1% and 96% identical to BjDGAT2 across 1512 coding region and 503 overlapping deduced amino acids respectively. These proteins were alkaline in nature (pI, 8.5-8.6), having similar molecular size (56-57 kD), an N-terminal hydrophilic segment and 9 transmembrane segments. Diacylglycerol/phorbol ester-binding motif (HKWXXRHXYXP) and acyl CoA binding motif (FYXDWWN) required for binding of substrates remained conserved in these proteins. Expression of the two transcripts of DGAT and their role in oil biosynthesis can further be studied.

018439   Hamdy H S (Biology Dep, Faculty of Education, Ain Shams University, Roxy 11757, Cairo, Egypt) : Extracellular collagenase from Rhizoctonia solani: production, purification and characterization. Indian J Biotechnol 2008, 7(3), 333-40.

Potentiality of R. solani grown on Sabouraud-glucose-collagen medium to produce glycosylated metallo-proteinase with collagenolytic activity was optimized and maximum production (212.33 U/mL) was recorded after 108 h of submerged incubation (175 rpm) at pH 5.5 and 30°C of temperature. Two-step column chromatography technique on DEAE-cellulose and Sephadex G150 was adopted to purify the partially purified enzyme produced by ammonium sulfate (40%, w/v) precipitation. Yield of purification was 60.49% of the original activity with specific activity of 18064.7 x 103 U/mg protein and 18.72-folds of purification. The purified enzyme showed maximum activity at 40°C and pH 5 ,which was stimulated by ions of Ca, Co, Cu, K, Mg, Na or Zn and inhibited by ions of Fe and Hg. Metal composition of the purified enzyme revealed that it contains Ca²⁺ and Zn²⁺. Tm (midpoint of thermal inactivation) was recorded at 65°C and 55°C after 1 and 6 h of exposure, respectively and T_1/2 was found to be 7 and 5 weeks at 15°C and 4°C, respectively. Molecular mass, K_mg/mL, V_max of the enzyme were found to be 66± 4 kDa, 0.033 kDa and 0.28 U/mg min^-1, respectively. Activities toward gelatin and casein were also detected.

018438   Goyal S;Ramawat K G (Biomolecular Technology Lab, Botany Dep, M L Sukhadia University, Udaipur-313 001) : Increased isoflavonoids accumulation in cell suspension cultures of Pueraria tuberosa by elicitors. Indian J Biotechnol 2008, 7(3), 378-82.

Cell cultures of Pueraria tuberosa were established in modified Murashige and Skoog medium and challenged with yeast extract (YE), methyl jasmonate (MeJA) and salicylic acid (SA). Maximum isoflavonoids production was recorded at 48 h of YA incorporation at the stationary phase. 20 μM of MeJA and SA was most effective in isoflavonoids induction. Higher concentrations of these elicitors were negatively correlated with the isoflavonoids production. YE at 150 mg L^-1 was optimal for isoflavonoids production, yielding 10 mg L^-1 isoflavonoids, which was <126>20% higher over the yields at optimal concentrations of MeJA and SA. YA incorporation can be used as a trigger to induce high yield of isoflavonoids.

018437   Goyal D;Bhadauria S (Botany Dep, Raja Balwant Singh College, Agra-282 002) : In vitro shoot proliferation in Emblica officinalis var. Balwant from nodal explants. Indian J Biotechnol 2008, 7(3), 394-7.

Efficient protocol for in vitro shoot proliferation of Emblica officinalis var. Balwant has been developed by using nodal explants. In vitro shoot proliferation from nodal explants have been observed by using MS, B5 and WPM media. Amongst them, MS medium has been observed the best for shoot proliferation (63.3%), while minimum phenol leaching (63.3%) was observed on WPM medium. To enhance the shoot regenerative potential, explants were cultured on MS medium supplemented with various cytokinins, viz., BAP, Kn and TDZ. MS medium supplemented with BAP (4.44 μM/L) was observed the best among all cytokinins. As shoot proliferation depended upon the balance of cytokinins and auxins, and BAP was found ideal for shoot proliferation, additive effect of auxins - IAA, IBA and NAA was assessed at 4.44 μM/L conc. of BAP. Explants when cultured on MS + BAP (4.44 μM/L) with IBA (2.46 μM/L)) showed maximum percentage of shoots (85%) with minimum callus formation (15%). Higher concentration of auxins suppressed the caulogenesis and induced callus formation.

018436   Gayathri Elayidam U;Muraleedharen D (Zoology Dep, University of Kerala, Kariavattom, Thiruvanathapuram-695 581, Email: gayathri_elayidam@yahoo.com) : Identification and partial characterization of juvenile hormone esterase from cotton pest Dysdercus cingulatus. Indian J Biochem Biophys 2008, 45(2), 121-5.

Juvenile hormone esterase (JHE), a selective enzyme that hydrolyzes the methyl ester of insect juvenile hormone plays an important role in regulating metamorphosis in nymphs as well as reproduction in adults. Studies on JH degradation provide insight into the possibilities of physiological disruption in the insects. In the present study, the JH degrading enzyme, JHE from the cotton pest Dysdercus cingulatus (Heteroptera) is characterized. Electrophoretic analysis of haemolymph during various developmental stages showed the JHE bands prominent only on the final day of 5th instar nymph, and the esterase substrate specificity confirmed the presence of JHE isoforms. In an attempt to clone cDNA of JHE gene from the final instar nymphs, mRNA isolated from fat bodies was coupled with JHE gene-specific primers and the cDNA was synthesized using RT-PCR. The PCR amplified cDNA showed the presence of JHE isoforms in D. cingulatus.

018435   Gao Z Q;Meng C X;Wang G C;Ye N H (School of Life Sciences, Shandong University of Technology, 255049, Zibo, China, , Email: zq77232002@yahoo.com) : Isolation and characterization of photosystem II from the filamentous sporophyte of Porphyra yezoensis. Indian J Biochem Biophys 2008, 45(4), 244-9.

Thylakoid membranes were isolated and purified from diploid filamentous sporophytes of Porphyra yezoensis Ueda using sucrose density gradient ultracentrifugation (SDGUC). After thylakoid membranes were solubilized with SDS, the phtosystem II (PSII) particles with high 2, 6-dichloroindophenol (DCIP) photoreduction activity were isolated by SDGUC. The absorption and fluorescence spectra, DCIP photoreduction activity and oxygen evolution activity of the thylakoid membranes and PSII particles were determined. The polypeptide composition of purified PSII particles was distinguished by SDS-PAGE. Results showed that PSII particles of sporophytes differed from the gametophytes in spectral properties and polypeptide composition. Apart from 55 kDa D1-D2 heterodimer, CP47, CP43, 33 kDa protein, D1, D2, cyt b559 and 12 kDa protein were identified from PSII particles from sporophytes; a new 102 kDa protein was also detected. However, cyt c-550, 20 kDa, 14 kDa and 16 kDa proteins found in PSII particles from gametophytes were not detected in the sporophytes.

018434   El-Shahed K Y I;El-Diwany A I;Awad H M M (National Research Center, Chemistry of Natural and Microbial Products Department, Dokki-Cairo-Egypt) : Enhanced production of streptomycin and hydrolytic enzymes by Streptomyces griseus strains using different types of organic solvents and detergent compounds. Indian J Biotechnol 2008, 7(3), 341-8.

Different organic solvents and detergents showed profound effects on the production of streptomycin (SM), amylases and proteases by Streptomyces griseus strains. The maximum SM production (1128 and 780 mg/L) was obtained using Streptomyces NRRL-strain when ethanol (1.0% v/v) and Tween-80 (0.1%v/v) were added to the medium. Production of amylases was also enhanced using the strain with the addition of benzene at decline phase, n-butanol at the trophophase and isopropanol at the idiophase with values, 10.66, 8.28 and 8.05 U/mL for a-amylase (EC-3.2.1.1) and 6.28, 4.68 and 4.55 for b-amylase (EC-3.2.1.2), respectively. While the use of other solvents and detergents reduced the production of amylase specially activities by S. griseus DSM-40759, characterized by higher productions of a- and b-amylases in different growth phases especially at the death phase. Worthily, proteases secretion was only induced to 1.02 U/mL at the idiophase by culturing DSM-strain with the addition of Triton X-100 and SDS to the fermentation medium. Moreover, the solvents shifted the pH values in the acidic range by the two strains at different phases compared to the control.

018433   Eapen S;Kamble S;Suseelan K N (Nuclear Agriculture and Biotechnology Division, Bhabha Atomic Research Centre, Trombay, Mumbai-400 085, Email: eapenhome@yahoo.com) : Rhizofiltration of cadmium and lead by hairy root cultures of Brassica juncea L. and the influence of the metals on antioxidative enzymes. Pl Cell Biotechnol molec Biol 2007, 8(1-2), 73-8.

Plant hairy roots induced by Agrobacterium rhizogenes can be used as a model system to study the uptake of heavy metals and the influence of metals on the antioxidative enzymes. Hairy roots induced in Brassica juncea L. Czern and Coss. When used for remediation of cadmium and lead from solutions, 90% of Pb from solutions upto 1000μM and 60% of Cd from 1000μ solutions could be taken up within 2 hrs of treatment. Antioxidant enzymes are known to be important defence system of plants to metal stress and enhanced activity of catalase, superoxide dismutase and guiacol peroxidase enzymes was observed in hairy root at high concentrations Cd, while in the case of Pb, the increase in enzyme activity was seen only for catalase (CAT) and superoxide dismutase (SOD) and not for guiacol peroxidase.

4 illus, 18 ref

018432   Dubey H;Haider Z A (Coll of Biotechnol, Birsa Agric Univ, , Ranchi-834 006) : Stevia : the naturally occurring sweet plant. J Res-Birsa Agric Univ 2007, 19(2), 261-7.

Stevia is likely to become a major source of high potency sweetener for the growing world food market. It is imperative to convert Stevia from a wild plant to a plant suited to meet the huge demand of the industry. Therefore, efficient and fast mechanized production needs to be established for Stevia to address the need. Understanding the biology of the Stevia plants and biochemistry of the sweet glycosides are prerequisites for conversion of Stevia to a modern crop. The successful exploitation of this plant will ensure tremendous boost to the economy of farmers and also help in answering the medical issues related to a vast group of population. Protocol for mass propagation of Stevia are available which marks the embarking of the efforts put towards the successful exploitation of this plant.

1 illus, 1 table, 61 ref

018431   Dhiman M R (NO, Indian Agricultural Research Institute, Regional Station, Katrain, Kullu Valley-175 129, Email: mrarjun01@gmail.com) : Effect of scale position and growing medium on bulblet formation and growth during scale propagation of Lilium. J ornamental Hort 2007, 10(2), 119-21.

Experiment was carried out to investigate the effect of scale position and growing medium on bulblet formation and growth, when Lilium hybrid cultivar Grand Care was scale propagated. Maximum number of bulblets per scale (2.68) was recorded in inner scales. Sand produced highest number (2.61) of bulblets per scale. Weight (1.86g) and diameter (1.38cm) of bulblets were higher in inner scales. Maximum weight of bulblet (2.27g) in inner scale was recorded with sand + vermiculite. More number of roots (5.56) per scale were found in inner scale. Further, it was maximum (6.15) in vermiculite. More number of roots per scale (7.33) were observed with vermiculite. Root length (8.79cm) was highest in sand + vermiculite. Maximum root length (10.14 cm) in inner scale was recorded with sand + vermiculite.

3 tables, 9 ref

018430   Das N;Vimala R;Karthika P (School of Biotechnology, Chemical and Biomedical Engineering, VIT University, Vellore-632 014) : Biosorption of heavy metals-an overview. Indian J Biotechnol 2008, 7(2), 159-69.

During the last two decades, extensive attention has been paid on the management of environmental pollution causal by hazardous materials such as heavy metals. Decontamination of heavy metals in the soil and water around industrial plants has been a challenge for a long time. A number of methods have been developed for the removal of heavy metals from liquid wastes such as precipitation, evaporation, electroplating, ion exchange, membrane processes, etc. However, these methods have several disadvantages such as unpredictable metal ion removal, high reagent requirement, generation of toxic sludge, etc. Biosorption is a process, which represents a biotechnological innovation as well as a cost effective excellent tool for removing heavy metals from aqueous solutions. This article provides a selective overview of past achievements and present scenario of biosorption studies carried out on some promising natural biosorbents (algae, fungi, bacteria, yeast) and some waste materials which could serve as an economical means of treating effluents charged with toxic metallic ions.

018429   Das D K;Saini M;Devendra Swarup;Gupta P K (Centre for Wildlife, Indian Veterinary Research Institute, Izatnagar-243 122) : Comparison of nucleotide and amino acids sequence of Nilgai (Boselaphus tragocamelus) interleukin-18 (IL-18) with other ruminants. Indian J Biotechnol 2008, 7(2), 195-9.

Interleukin-18, a component of innate immunity with potent interferon-γ-inducing activity, is not well characterized in wild ruminants. In this study, IL-18 cDNA of Nilgai (Boselaphus tragocamelus) was cloned and characterized. The nucleotide sequence was 582 bp long and contained its entire open reading frame encoding 193 amino acid residues. The sequence analysis indicated seven nucleotide substitutions with reported buffalo sequence showing 98.5% similarity, 97.8% with cattle and 97.6% with sheep. The Nilgai IL-18 cDNA included a putative cleavage site of IL-1β-converting enzyme (ICE) and IL-1 signature-like sequence identified in human and cat IL-18 cDNA. Both nucleotide as well as amino acid sequence similarity shows that the cloned sequence was closer to buffalo IL-18 sequence.

018428   Dada R;Kumar R;Shamsi M B;Tanwar M;Pathak D; Venkatesh S;Kumar M;Singh H;Singh K;Aron M;Kumar R;Singh G;Sharma R K;Gupta N P (Molecular Reproduction and Genetics Lab, Anatomy Dep, All India Institute of Medical Sciences, New Delhi-110 029, Email: rima_dada@rediffmail.com) : Genetic screening in couples experiencing recurrent assisted procreation failure. Indian J Biochem Biophys 2008, 45(2), 116-20.

Infertility is a major health problem affecting about 10-20% of couples in the reproductive age group. Male factor is assumed to be responsible in about 50% cases of infertility. The origin of reduced testicular sperm function is unknown in about 50-70% of cases and for such couples assisted reproduction techniques (ART) are a boon. Male infertility is often due to poor semen quality and may be associated with genetic defects. ART has revolutionized management of infertility and intracytoplasmic sperm injection (ICSI) is the ART procedure of choice in 60-80% cases. Despite major technological advancements and professional expertise in ART, the success rate and carry-home live birth rate of ICSI is low (18-25%). This study was aimed to understand the genetic etiopathology of recurrent ART failure. For this, 110 couples with 3 or more failed ART cycles were recruited. A detailed history was taken and only idiopathic ART failure cases were enrolled for this study. They were subjected to cytogenetic and Yq microdeletion analysis. Genetic abnormalities were detected in 19 couples. Since a large number (18.2%) cases harboured genetic abnormalities, it is important for all couples opting for ART to undergo a thorough genetic analysis to prevent recurrent emotional, physical and financial stress.

018427   Chi Z;Li J;Wang L;He S;Wang X (UNESCO Chinese Center of Marine Biotechnology, Ocean University of China, Yushan Road, No.5, Qingdao 266003, China, Email: zhenming@sdu.edu.cn) : Pichia pastoris INO1 gene expression affects intracellular maltase activity and MAL1<. Indian J Biochem Biophys 2008, 45(3), 174-8.

Examins the influence of INO1 gene expression on intracellular maltase activity and MAL1⁺ gene expression in these two transformants. The highest specific maltase activity was observed when Sch.p944 was grown in the synthetic medium containing initial 2.0% (w/v) glucose, whereas glucose repression of maltase activity occurred when Sch.p1025 was cultivated in the same medium containing glucose more than initial 0.5% (w/v), suggesting that synthesis of the maltase was derepressed in Sch.p944. Also, higher mRNA encoding intracellular maltase activity was found in Sch.p944 than that in Sch.p1025 after 32 h of cell growth in the media containing initial 2.0% glucose, indicating that the derepression occurred at the transcriptional level. Furthermore, higher phosphatidylinositol (PI) content was detected in Sch.p944 cells grown in the medium containing initial 2.0% (w/v) glucose for 32 h than in Sch.p1025 cells grown under the same conditions, indicating that PI might be involved in derepression of MAL1⁺ gene expression and intracellular maltase activity.

018426   Chandran R P;Potty V P (NO, Central Tuber Crops Research Institute, Thiruvananthapuram-695 017) : Induction of hairy roots through the mediation of four strains of Agrobacterium rhizogenes on five host plants<. Indian J Biotechnol 2008, 7(1), 122-8.

Induction of hairy roots by four strains of Agrobacterium rhizogenes- ATCC 15834, A4, WC and WR were studied in five plants, Ipomoea batatas, Solenostemon rotundifolius, Vigna vexillata, Pachyrrhizus erosus and Canavalia species. Among the five plants selected for transformation and induction of hairy roots, P. erosus was found resistant to all the four bacterial strains. Similarly, one strain, WR also failed to induce hairy roots in all the plants. However, all the strains exhibited good growth dominated by 15834 grown in YEB medium. Hairy roots were induced from the cotyledons, hypocotyls, stem cuttings and in vitro plants of I. batatas through the transformation of 15834 and A4 strains. S. rotundifolius and V. vexillata were susceptible to the strains of A4, 15834 and WC. Canavalia sp. was resistant to WR and WC strains, but was susceptible to A4 and 15834. It was for the first time that hairy roots were initiated from S. rotundifolius, V. vexillata and Canavalia sp. The variation observed in the time of induction of hairy roots (incubation period) by a single strain (15834) in different plant species, suggests that the plant has also a definite role in determining the incubation period. Among the four strains of A. rhizogenes, 15834 was found to be the most efficient in transformation and initiation of hairy roots, with the shortest minimum incubation period and dominant growth in YEB medium. A. rhizogenes is a well known plant pathogen, which produces "hairy root disease" in susceptible plants. On modified MS medium, cotyledon explants were superior to hypocotyls. The hairy roots transformed by A. rhizogenes strain 15834 on I. batatas, V. vexillata and Canavalia sp. were also morphologically different.

018425   Chakravarty A K;Yasmin H (Immunology & Cell Biology Lab, Zoology Dep, School of Life Sciences, University of North Bengal, Siliguri 734 013, Email: prof_ashim_chakravarty@rediffmail.com) : Activation of cell mediated immune response and apoptosis towards malignant cells with turmeric treatment in murine model. Indian J Biochem Biophys 2008, 45(1), 23-9.

The effect of ethanolic turmeric extract (ETE) on murine lymphocytes vis-…-vis tumor cells was studied, in terms of its ability to activate lymphocytes and to induce apoptosis in tumor cells. Degree of activation and proliferation of lymphocytes treated with ETE was analyzed in terms of blastogenesis, DNA synthesis through ³H-thymidine incorporation and cell cycle analysis by flourescence activated cell sorter (FACS). FACS analysis was also carried out to observe the proliferation as well as apoptosis of tumor cells. Morphological condition of both the cell types in presence of ETE was examined by scanning electron microscopy (SEM). Cytotoxic capability of ETE-treated effector T lymphocytes towards tumor cells was judged in vitro by ⁵¹Cr-release assay and the growth of tumor in situ. ETE stimulated murine lymphocytes towards blastogenesis and synthesis of DNA, as revealed by increased incorporation of ³H-thymidine. FACS indicated that the lymphocytes were driven towards mitotic cycle by activating G2-M transition. In the same count, the tumor cells mostly remained accumulated in the G₂-M phase, and thus mitotically arrested. Scanning electron photomicrographs revealed the blastoid transformation of lymphocytes and ETE-induced apoptotic condition of tumor cells. Furthermore, ETE-treated T cells were cytotoxic towards tumor cells in vitro, as shown by ⁵¹Cr- release assay. ETE administered intravenously or orally could delay the onset and growth of tumor, and thus prolonged the life span of the tumor-bearing host. The investigation suggests potential of turmeric both to destroy the malignant cells directly and via activation of the host's cellular immunity.

018424   Bhatia S;Patil S S;Sood R;Dubey R;Bhatia A K; Pattnaik B;Pradhan H K (High Security Animal Disease Laboratory (HSADL), Indian Veterinary Research Institute, Bhopal-462 021) : Prokaryotic expression of a 750 bp capsid region of bovine immunodeficiency virus gag gene and development of a recombinant capsid (p26) protein based immunoassay for seroprevalence studies. Indian J Biotechnol 2008, 7(1), 50-5.

750 bp DNA fragment of the gag gene, coding for capsid (p26) protein of bovine immunodeficiency virus (BIV), was cloned in pQE32 vector and expressed as 6' His tagged fusion protein in Escherichia coli. The concentration of affinity purified His tagged capsid protein was 5 mg/mL and its yield was 3.5 g/L of induced culture (E. coli). The recombinant capsid protein of BIV was found to be immunologically reactive with a reference positive serum. Using the purified capsid protein, an indirect ELISA was standardized to test sera of cattle and buffalo for carrying out sero-surveillance of BIV. Of 672 animals tested by capsid based ELISA, 162 were positive and 510 were negative, giving an overall prevalence of 24% in India. In conclusion, the recombinant capsid based indirect ELISA was found suitable to study BIV antibody status. To our knowledge, this is the first seroprevalence study of BIV infection in India.

018423   Bhatia S;Arora R (Animal Genetics Div, National Bureau of Animal Genetic Resources, Karnal-132 001) : Genetic diversity in Kheri - a pastoralists developed Indian sheep using microsatellite markers. Indian J Biotechnol 2008, 7(1), 108-12.

The Kheri sheep were analyzed using 25 ovine microsatellite markers proposed by Food and Agriculture Organization, International Society for Animal Genetics (FAO-ISAG). All the used microsatellite markers amplified well and exhibited polymorphism. Wide range of variability depicted by number of observed alleles from 2 (BM6506, CSSM47 and OarCP20) to 10 (CSSM31 and OarJMP29), observed heterozygosity from 0.087 (OarCP20) to 1.000 (OarHH35), expected heterozygosity from 0.083 (OarCP20) to 0.828 (BM1314) and Polymorphism Information Content (PIC) from 0.079 (OarCP20) to 0.806 (BM1314) supported the utility of these microsatellite loci in measurement of genetic diversity indices in Indian sheep too. The mean number of observed and effective alleles was 5.3 and 3.3, respectively. The average observed heterozygosity values (0.582) compared to the average expected heterozygosity values (0.651) did not show significant differences in the selected population (p<62>0.05), which suggested random mating in Kheri. The allele diversity (average number of alleles per locus) and gene diversity (average expected heterozygosity) reflected high levels of genetic variability in Kheri sheep. Within population inbreeding estimate (Fis) was significant (p<60>0.05) and equal to 12.8%. The deficit of heterozygotes may be partly explained by Wahlhund effects at level of sampling (i.e. sampling at random from the whole population), although the main factor that appears to have provoked this lack of heterozygotes may be attributed to consanguinity that has resulted in accumulation of inbreeding in the breeding groups. This study contributes to the knowledge of genetic diversity of Kheri-the black brown faced meat, carpet wool sheep developed by migratory pastoralists in India.

018422   Beshay U (Bioprocess Development Dep, Genetic Engineering and Biotechnology Research Institute, Mubarak Cit, New Bourg El-Arab City, Alexandria, Egypt) : Oxygen transfer conditions in the production of rainbow trout growth hormone (rtGH) by Escherichia coli. Indian J Biotechnol 2008, 7(2), 225-9.

Batch cultures of Escherichia coli pRE1-rtGH 121/MZ1 were carried out at different oxygen transfer rates (OTR) enhanced by the increase of agitation and/or aeration rates. The growth kinetics of E. coli was investigated in complex medium under batch cultivation in 3 L fermenters. The agitation rates influenced both cell growth and rainbow trout growth hormone (rtGH) production in the bioreactor. It was found that increasing the agitation speed had a positive effect on the production of rtGH. Moreover, varying the airflow rate (vvm) in the fermentor under constant drive speed, cell growth and rtGH production were greatly influenced. In the production of rtGH by E. coli, high aeration rates were found to be essential for good yields of recombinant protein. Agitation speed and aeration rate could affect dissolved oxygen concentration, which in turn affected cell growth and rtGH production. Increased aeration rates induced higher rtGH production, with the highest concentration of 0.957 g/L obtained at 4.0 vvm, within 23 h. The highest volumetric productivity for rtGH of 0.042 g/L/h was obtained at both 1000 rpm and 4.0 vvm.

018421   Banerjee B D;Chakraborti A;Suke S G;Ahmed R S;Tripathi A K (Environmental Biochemistry and Immunology Lab, Biochemistry Dep, University College of Medical Sciences and G.T.B. Hospital (University, Dilshad Garden, Delhi-110 095, Email: banerjeebd@hotmail.com) : Xenobiotic-induced Immune Alterations: Implications in Health and Disease. Indian J Biochem Biophys 2008, 45(1), 7-15.

Immune function may be significantly altered following occupational, inadvertent or therapeutic exposure to chemically diverse xenobiotics. The environmental chemicals like pesticides, halogenated hydrocarbons, polychlorinated dibenzofurans, organic solvents, asbestos, silica, heavy metals etc. may interact with both cellular and humoral components of the immune system which can result in altered immune status that in turn may lead to decreased resistance to infection, certain forms of neoplasia or in some cases exacerbate allergy or autoimmunity. Recent advances in pharmacogenomics and toxicogenomics have contributed a lot to delineate the mechanism of interaction of xenobiotics with the biological system at the cellular and molecular level. However, detection of immune changes on exposure to immunotoxic agents is highly complex, especially in humans due to several confounding factors like age, sex, race gender, co-existence of disease, food habits, smoking etc. Thus, establishing a quantitative relationship between immunotoxicological data and risk assessment, following xenobiotic exposure is still a challenge. Reviews the immune alterations caused by exposure to variety of xenobiotics, and their implications in health and disease.

018420   Badgujar S B;Kosalge S B (Dep of Biotechnology, S S B T's College of Engineering and Technology, Bambhori, Jalgaon, Maharashtra, Email: sham83badgujar@yahoo.co.in) : Preliminary phytochemical and antibacterial screening of Cassia Australis Linn. J Sci Pharm 2007, 8(3), 99-103.

Aqueous, hydro alcoholic, methanol, petroleum ethr and chloroform extracts of dry powdered leaves of Cassia australis Linn. were subjected to preliminary qualitative chemical investigations. Carbohydrates, proteins, steroids, flavonoids, tannins and glycosides were found to be present. Antibacterial activity of theses extracts were evaluated against Bacillus subtilis, Pseudomonas auriginosa, Staphylococcus aureus, Proteus vulgairs, Escherichia coli an Shigella sonnei by agar cup - plate method using nutrient agar medium. All the extracts of C. australis have shown significant antibacterial activity in comparison with the standard antibiotic drug. This plant can be a promising source for the isolation of active phytoconstituent having anti-infective property against various pathogenic microorganisms.

5 tables, 8 ref

018419   Babu V S;Narasimhan S;Nair G M (Botany Dep, University of Kerala, Kariavattom-695 581) : Enhanced accumulation of triterpenoids and flavonoids in cell suspension cultures of Azadirachta indica (A. Juss.) with an extended stationary phase. Indian J Biotechnol 2008, 7(2), 270-2.

Accumulation of three triterpenoids (azadirachtin-A, nimbin and salannin) and two flavonoids (quercetin and kaempferol) was analyzed in cell suspension cultures of Azadirachta indica A. Juss. By feeding cells with a sterile solution of sucrose, it was possible to sustain viability of cells in batch cultures without subculturing for 60 d. Such long-term cell suspension cultures exhibited an extended steady stationary phase which facilitated enhanced accumulation of azadirachtin-A, nimbin, salannin, quercetin and kaempferol. 40% of the cells were viable towards the end of 60 d of incubation. Maintaining cells for longer time under stationary phase facilitated transformation of cells from low productivity to high productivity. The cells in stationary phase could serve as a starting material for the selection of elite cell lines and for bioreactor cultures for large-scale production of these compounds.

018418   Awasthi A K;Pradeep A R;Srivastava P P; Vijayan K;Vineet Kumar;Urs S R (Seribiotech Research Lab, Central Silk Board, Carmelaram Post, Kodathi, Bangalore-560 035) : PCR detection of densonucleosis virus isolates in silkworm (Bombyx mori) from India and their nucleotide variability. Indian J Biotechnol 2008, 7(1), 56-60.

Densonucleosis virus (DNV) is one of the pathogenic viruses of the commercially valuable silkworm, Bombyx mori. It causes flacherie disease, mostly as combined infection with other pathogens like bacteria, which accounts for the significant loss of cocoons in sericulture. Two isolates of DNV from B. mori, DNV1 and DNV2 have been previously identified on the basis of their sequences. After infection with purified isolates of DNV in some commonly used silkworm strains, viz. Nistari, C'nichi, NB1 and Guangnnong Marked, the polymerase chain reaction (PCR) was conducted using DNV1 and DNV2 primers. DNV1 primers generated a distinct profile in the B. mori strains, whereas DNV2 produced single monomorphic band in all the screened strains. Sequence of one of the prominent fragments generated by the DNV1 primer exhibited very high degree of nucleotide variability from that of Japanese DNV1 isolate, but the sequence of DNV2 showed near to complete similarity. Besides, the study demonstrates that PCR technique could be used to diagnose the DNV presence/absence in silkworm strains without sacrificing the larvae and the results could be used in breeding programmes.

018417   Awasthi A K;Kar P K;Srivastava P P;Rawat N; Vijayan K;Pradeep A R;Urs S R (Seribiotech Research Lab, Central Silk Board, Carmelram Post, Kodathi, Bangalore-560 035) : Molecular evaluation of bivoltine, polyvoltine and mutant silkworm (Bombyx mori L.) with RAPD, ISSR and RFLP-STS markers. Indian J Biotechnol 2008, 7(2), 188-94.

Mulberry silkworm (Bombyx mori L.), the most important silk producing insect, exhibits wide diversity in morphological and biometric characters. Characterization of vast genetic resources based on the morphological and quantitative traits is not solely dependable as the phenotypic traits are influenced by environment. In order to study genetic relatedness of the selected and varied genotypes, six each of the bivoltine, polyvoltine silkworm accessions and mutant stocks, were studied with RAPD, ISSR and RFLP-STS markers. Twelve RAPD primers generated 172 markers of which 161 were polymorphic, generating 93.60% polymorphism. Pair-wise genetic divergence varied from 0.209 between Mysore Princess and Rong Diazo to 0.588 between Boropolu and TMS-35. The ISSR primers generated 156 markers of which 132 were polymorphic thus generating 84.62% polymorphism. The pair-wise genetic diversity among the genotypes varied from 0.189 between Rong Diazo and BL-23 to 0.438 between MU-10 and TMS-35. Similarly, 10 RFLP-STS primers produced a total of 69 bands, out of which 53 were polymorphic thus realizing 75.6% polymorphism. Genetic distance varied from 0.242 between Nistari-M and BL-23 to 0.730 between Fengshong and TMS-17. On clustering with UPGMA and principal component analysis (PCA), RAPD and ISSR markers clearly discriminated the bivoltines and multivoltines and a multivoltine Tamil Nadu white occupied the positions among bivoltines, since it has bivoltine parentage. Boropolu, an original land race from North East India, and Feng shong, a Chinese silkworm strain, also showed a closer genetic relationship.

018416   Arya D;Patni V;Uma Kant (Plant Pathology, Tissue Culture and Biotechnology Lab, Botany Dep, University of Rajasthan, Jaipur-302 004) : In vitro propagation and quercetin quantification in callus cultures of Rasna (Pluchea lanceolata Oliver & Hiern.). Indian J Biotechnol 2008, 7(3), 383-7.

Protocol for micropropagation of Pluchea lanceolata, an important medicinal herb was developed. Leaf explants obtained from field grown plants when tested for callus induction on Murashige and Skoog's (MS) medium, supplemented with NAA in combination with BAP, produced the best callus. Maximum number of multiple shoots from the callus (26.6±0.67) was obtained on MS medium supplemented with BAP (1.0 mg/L) and Kn (1.0 mg/L). More or less uniform elongation of multiple shoots was obtained on MS medium with lower concentrations of cytokinins, i.e., BAP (0.25 mg/L) and Kn (0.5 mg/L). Further elongation and profuse rooting were achieved when the well-grown shoots were cultured on half strength MS medium supplemented with IBA (1.0 mg/L). The regenerated plantlets were hardened and established at 70% survival rate in pots. The bioactive secondary metabolite, quercetin, was isolated from callus tissues of different age groups and its identification and confirmation was carried out by the colour reaction, TLC behaviour, IR spectrum and HPLC techniques. Maximum quercetin content (0.23 mg/g dry wt of tissue) was obtained in 6-wk-old callus tissues.

018415   Arutchelvi J;Sudhakar M;Arkatkar A;Doble M; Bhaduri S;Uppara P V (Biotechnology Dep, Indian Institute of Technology, Madras, Chennai-600 036) : Biodegradation of polyethylene and polypropylene. Indian J Biotechnol 2008, 7(1), 9-22.

Polyethylene and polypropylene are the two polyolefins with wide ranging applications. They are recalcitrant and hence remain inert to degradation and deterioration leading to their accumulation in the environment, and, therefore creating serious environmental problems. Biodegradation of these two polymers under in vitro conditions is reported. An attempt has been made to cover the mechanism of biodegradation, the various bacterial and fungal organisms that have been reported for the same, methods adopted for the studies and different characterization techniques followed to measure the extent of degradation.

018414   Arun Kumar N;Lakshmi Narasu M;Zehr U B;Ravi K S (NO, Mahyco Research Center, Jalna-Aurangabad Road, Dawalwadi, Post Box No. 76, Jalna-413 203) : Molecular characterization of Tobacco streak virus causing soybean necrosis in India. Indian J Biotechnol 2008, 7(2), 214-7.

A virus isolate infecting soybean (Glycine max L.) with characteristic symptoms of necrosis was collected from various locations of Maharashtra, India. The virus was detected as Tobacco streak virus (TSV) by direct antigen-coating-enzyme-linked immunosorbent assay (DAC-ELISA) using TSV specific antiserum. ELISA positive TSV soybean isolates upon mechanical transmission onto indicator hosts, viz. Vigna unguiculata cv. C-152, Glycine max and Nicotiana tabacum cv. Xanthi, produced characteristic local necrotic lesions on primary inoculated leaves, followed by systemic infection. Coat protein (CP) gene from a representative soybean isolate (TSV-SB) was amplified using TSV CP specific primers. The amplicon of <126>750 bp was cloned and sequenced. The CP gene consists of 717 nucleotides, potential of coding a polypeptide of 238 amino acid residues. The CP gene of TSV-SB isolate shared 98.3 to 99.3% nucleotide and 96.6 to 98.3% amino acid sequence homology with the TSV isolates characterized from India. With TSV-WC (White clover isolate from America, type strain) and TSV-BR (Soybean isolate from Brazil), TSV-SB isolate shared 88.7% and 79.2% amino acid sequence homology, respectively. Phylogenetic tree constructed based on amino acid sequence analysis showed a close clustering of TSV-SB isolate with other TSV isolates from India than with the American and Brazilian strains. Based on CP gene sequence analysis, it is concluded that the TSV-SB isolate is a strain of TSV that is prevalent in India. This is the first report of molecular characterization of TSV infecting soybean from India.

018413   Anupama P M;Ayyanna C;Ramana T (Biotechnology Dep, ANITS, Visakhapatnam) : Optimization of malting conditions of Pearl millet [Pennisetum glaucum (L.)R.Br.] using Box-Behnken design. Indian J Biotechnol 2008, 7(3), 361-5.

In the study, malting conditions of Pearl millet [Pennisetum glaucum (L.)R.Br.] were optimized using Box-Behnken design. The three variables chosen were gibberlic acid (X₁), temperature (X₂) of malting and days of germination (X₃) for optimal yield of α-amylases and proteases in the malt. A total of 15 runs by combination of each factor were carried out and contribution of each factor was established. The t-values and p-values obtained revealed that all the three variables significantly contribute to the formation of both enzymes during malting. Optimal conditions required for the formation of maximal expression of α-amylases (323.03 units/g of malt) as provided by the model are as follows: gibberlic acid 1.053 ppm, temperature to be maintained at 32.98°C and the malt must be obtained at the end of 3.68 d. The set of conditions maintained to get an optimal protease yield (32.968 units/g of malt) are: gibberlic acid, 1.0743 ppm; temperature, 33°C; and the malt must be obtained at the end of 3.4 d (78 h). Following these conditions, malt with α-amylase and protease activities of 320 units and 31.24 units per g, respectively were obtained.

018412   Anjana Kumari;Bhat R S;Kuruvinashetti M S (Biotechnology Dep, University of Agricultural Sciences, Dharwad-580 005) : Agrobacterium-mediated genetic transformation of BPT-5204, a commercially growin Indica rice variety. Karnataka J agric Sci 2007, 20(4), 845-7.

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018411   Ananda Krishna K;Sambasiva Rao K R S (Center for Biotechnology, Acharya Nagarjuna University, Guntur-522 510) : Stem cell based therapy to restore nearly normal hearing. Indian J Biotechnol 2008, 7(2), 178-82.

Hair cells with stereocilia are the sensory receptors of the inner ear that are located in the organ of corti of the cochlea, involved in detecting sound, and are connected with the nerve fibers that stretch from the inner ear to the brain. These hair cells convert sound information to electric signals, which are then sent to the higher brain centers. In humans, hair cell damage results in permanent hearing impairment; whereas, birds have the capacity to rebuild a damaged inner ear and various studies have demonstrated hair cell regeneration. Recently, stem cell research has triggered many programmes around the world to examine the possibility for regenerating the hair cells from embryonic/adult stem cells. The important spin-off of the research would be to find signaling mechanisms that regulate hair cell regeneration to restore nearly normal hearing in humans. The current review focuses on stem cell-based therapy with particular emphasis to summarize the recent progress.