Absract Archive   March 2010 Research Methodology Series Literature Review for Research Projects in Science and Technology Introduction The goal of any research project is to increase the 'knowledge base' in a particular area of science and to contribute to the development of the chosen area. The interest and initiative to conduct a particular line of research stems from ones' own specialization combined with the urge to further contribute the developing knowledge in that area of science. The specific area of science in which a research project is undertaken, requires a thorough understanding of earlier work done. Most research areas of potential knowledge generation will have many groups globally working constantly into the finer nuances simultaneously. Needless to say, many years of knowledge seeking and dissemination of results thus obtained in a globally accepted method is a powerful way of communicating and sharing scientific findings. A thorough understanding and assimilation of previous work done in a particular area will give a strong direction and focus to concentrate ones' resources for the knowledge towards progress. Also, this gives an overall understanding of the research assignment in hand and gaps in the field along with areas that need immediate attention. Review of literature is a comprehensive manuscript that incorporates all possible hitherto published work with easy reference citations and listings. The body of review can be constructed on major headings such as an Introduction to the area of work, the materials, methods (protocols), data and results as obtained previously. A comprehensive compilation and analysis of these should be made to arrive at a discussion which will form an important part of the review. While a compilation of all the protocols adopted will give an insight towards the experimental approach, such an exercise of the compiled results will give an idea as what to expect from the suitable modified/adopted protocols. Needless 'trial and error' experimental methodology along with the coupled resource utilization can be avoided which is very important to the sustenance of peak motivational levels to all involved in a project. A simpler or alternate methodology can be identified and adopted as also to develop novel protocols by a thorough understanding of a review of the methodology adopted hitherto. Identification of common errors, obtaining relevant data and analysis of the acquired data becomes easier with a thorough review of literature. A mere compilation of existing data will not make a proper review; rather an assimilated summary that gives the 'trend' and contributes to a discussion towards the future direction is more aptly a proper review. Also, this is useful for meaningful interpretation and infer the data/results obtained that will enrich the existing knowledge in a particular area of science. Author: M. Ravi For Correspondence: : E mail : maddalyravi@hotmail.com Research Article Effect Of Probiotic Bacteria In Indian Major Carp, Catla Catla (hamilton, 1822) On Growth Performance And Biochemical Parameters Abstract Lactobacillus plantarum and Bacillus megaterium was antagonized Aeromonas hydrophila with inhibition zone of 5 and 6 cm diameter, respectively. Both probiotics (L.plantarum & B.megatreium) were added to artificial basal diet with 35% crude protein to evaluate their efficacy on the growth-performance, Biochemical analysis and survival rate. Two hundred and forty Catla catla (Hamilton, 1822) fish with average body weight of 5.85 – 6.10 g/fish were equally divided into four treatments of triplet replicates. T1 was given basal diet (control), T2 was given basal diet with L.plantarum, T3 was given basal diet with B.megatreium, T4 was given basal diet with both L.plantarum & B.megatreium. Fishes were fed twice daily at the rate of 3% of their live body weight, for six days a week during the experimental periods (60 days). Fish were challenged by Aeromonas hydrophila (3% of total body weight i.e. 107 ± 1 cells g – 1) via feed and kept for 15 days. The best growth rate, feed utilization and survival rate were noticed with T2. Combination of two probiotics (L.plantarum & B.megatreium) enhanced the fish growth and health. It is recommended to use of combination of L.plantarum & B.megatreium as a probiotic bacteria in-vivo in aquaculture. Key words: Probiotic bacteria, Growth performance, biocontrol agent, Catla catla (Hamilton, 1822), Feed conversion ratio Authors: D. Ravi1*, R. Parthasarathy2, T. V. Logaraj3 For Correspondence: E Mail : dravi_botany@hotmail.com Research Article Optimization Of Culture Conditions For Alkaline Protease Production By Bacillus Subtilis Ncim 2063 In Submerged Fermentation Abstract Alkaline Protease production was carried out by employing Bacillus subtilis NCIM 2063. The strain was screened for protease production and initially the strain produced 0.47 units of protease per ml when grown in culture broth that contained glucose as carbon source, incubated at 35°C in an orbital shaker for 48 h. The most important process parameters were optimized i.e, temperature 37 degrees C, pH 8, incubation period 48h, inoculum size 1%. Among different additives supplemented, xylose, molasses and casein showed enhanced protease production. On the other hand amino acids, surfactants and trace elements supplemented to the fermentation medium showed a negative influence on alkaline protease production. The fermentor volume of 250 ml was found to be the best for maximum enzyme production. After optimization the strain produced 1.33 units of protease per ml of culture broth which is about 2.83 times more in the present study. Key words: Optimization, alkaline protease, Bacillus subtilis, submerged fermentation. Authors: S. Mrudula* and R. Nethravathi For Correspondence: E-mail : somamrudula@hotmail.com Research Article Evaluation Of In vivo Antioxidative Efficacy Of Protein Fraction Of Terminalia Catappa Leaves Using Swiss Albino Mice And Its Characterization Abstract Terminalia catappa Linn (Combretaceae) is found in the warmer parts of India. The various extracts of leaves and bark of the plant have been reported to be anticancer, anti-HIV reverse transcriptase, hepatoprotective, anti inflammatory, anti-hepatitis and aphrodisiac. Our present investigation concerns about the in vivo antioxidative role of protein fraction of Terminalia catappa in Swiss albino mice and its characterization. The proteins were precipitated by ammonium sulphate fractionation. These fractions were then estimated to choose the protein fraction having maximum protein content and DPPH assay was used to find the ED50 concentration of the protein to be used in further studies. The antioxidative effect of the protein fraction of Terminalia catapppa was evaluated in liver homogenate of Swiss albino mice. The proteins were characterized by PAGE and SDS-PAGE. More protein content was observed in 70% ammonium sulphate protein fraction and its ED50 concentration was found to be 40µg. This protein fraction possessed potent antioxidative role as evidenced from the significant increase in the activities of enzymic antioxidants, levels of non enzymic antioxidants and reduction in lipid peroxide levels in mice treated with protein fraction than that of mice treated with vehicle controls and silymarin, a standard reference antioxidant. This protein fraction was characterized by native PAGE and SDS PAGE and this reveals the presence of two monomeric proteins with the molecular weight 97 KD and 43 KD. From the present study it was concluded that this protein fraction of Terminalia catappa can be recommended as an antioxidative agent to fight against the degenerative diseases such as cardiovascular diseases, atherosclerosis, diabetes and cancer. Key words: DPPH assay, ED50 concentration, antioxidative, PAGE, protein Authors: J. Sivaprabha* and S.Annapoorani For Correspondence: E-mail : jsivaprabha@gmail.com Research Article HPTLC Separation Of Antibacterial Compounds From Perna Viridis And Portunus Sanguinolentus And Its Activity Tested Against Common Bacterial Pathogens Abstract Marine invertebrates were identified as Portunus sanguinolentus (three spotted crab) and Perna viridis (Green mussel). Crude extract from both animals was tested against common laboratory pathogens Escherichia coli, Klebsiella pneumonia and Staphylococcus aureus. The result of E.coli , K.pneumoniae and S. aureus in ethanol and methanol extraction of Perna viridis was 4.88, 4.26, 5.74 and 5.33, 4.64, 4.97 and Portunus sanguinolentus 5.32, 4.66, 6.53 and 7.45, 5.23, 5.66 respectively. Further high performance thin layer chromatography separated 5 different compounds from both the animal extracts. Among compounds 3 and 5 more active against common laboratory pathogens. When compared to the crude extracts the purified component showed a high antibacterial activity against the tested pathogens. In addition to both active compounds from marine invertebrates were sensitive to proteinase K and heat treatment. Thus the active molecules are most likely of peptides. The compounds from the animals were tested with human RBC thus they posses no hemolytic activity. This study showed the possibility of utilizing marine invertebrates as new promising pharmaceutical therapeutants which could be used in pharmaceutical industries. Key words: Marine invertebrates, HPTLC, Solvent, pathogens. Authors: A.Maripandi1,2* L. Prakash1 Ali A. Al-Salamah2 For Correspondence: Email address: drmari34@gmail.com Research Article Utilization Of Visceral Organ Waste Of Catla (Catla Catla) For The Isolation, Purification And Partial Characterization Of Protease Abstract Protease is an industrially important enzyme isolated from various sources. Its isolation from the visceral organ waste of Catla, showed the highest specific activity in 50-60 % ammonium sulphate precipitation. The change in increase of specific activity from 2.2 to 3.5 U/mg was noted with the dialyzed fraction. Further purification on Sephadex G-100 column revealed an increase of specific activity from 3.5 to 3.7 U/mg. The molecular weight of the enzyme was found to be 28.0 KDa. On Native PAGE it was found to be 48.0 KDa, which indicated the multimeric nature of the isolated protein. The optimum pH and temperature of the isolated enzyme was found to be 8 and 60°C respectively. Keywords: Catla, alkaline protease, fish waste, enzyme. Authors: Shobana.A* and Anitha Subash For Correspondence: Email id: shobana_a2001@yahoo.com   Research Article A Novel Approach In Cloning And Expression Of A Human Chemokine Gene In E. coli (BL21 DE3* Cells) Abstract Chemokines are important chemical messengers between cells of the immune system. They also play a critical role in many pathophysiological processes such as allergic responses, infectious and autoimmune diseases, angiogenesis, inflammation, tumor growth, and hematopoietic development. In this study, the human chemokine gene is amplified and digested with Nde I and EcoR I sites. The vector pET-20b(+) was also restricted with same endonucleases and further ligated with the target gene in a molar ratio of 1:10. It was transformed into DH5α strain to test the recombinants by performing restriction digestion analysis and PCR screening. Then the recombinant was transformed into BL21-DE3* cells of E. coli and subjected to SDS-PAGE for the expression study. The cloning of a gene of eukaryotic origin into a prokaryotic host has more significance in an economical production of the protein-Chemokine and may become important target for pharmacologic intervention in a wide variety of human diseases. Keywords: Human Chemokine gene, Restriction enzymes- Nde I and EcoR I, pET-20b(+) vector, BL21 DE DE3* Cells, PCR, SDS-PAGE. Authors: K. Jaianand1 * and N.Saravanan2 For Correspondence: Email: jaianand.k@lifecellinternational.com Short Communication “Detection Of Philadelphia Chromosome And BCR-ABL Gene In The Bone Marrow Cells Of Patients With Chronic Myeloid Leukemia” Abstract Chronic myelogenous leukemia (CML) is a pleuripotent stem cell disorder characterized by proliferation and accumulation of mature myeloid cells and their progenitors. During this study, a total of 25 patients (17males and 8 females with cytopathologically confirmed diagnosis of chronic myeloid leukemia (CML), were selected. Cytogenetic analysis for the detection of the Ph chromosome was carried out employing direct chromosome preparation from bone marrow. Chromosome banding was performed employing GTG banding Method and Karyotypes were prepared according to International System for human Cytogenetic Nomenclature. For the detection of BCR-ABL fusion transcripts Reverse transcription Polymerase chain reaction (RT-PCR) was employed. Conventional cytogenetic analysis of the 25 CML patients demonstrated the presence of Philadelphia chromosome formed as a result of translocation between 9 and 22 in 19 patients (76%) and 3 patients (12%) were demonstrated to be Ph negative cytogenetically and samples of 3 patients could not be analyzed because of poor quality of metaphases. But however molecular analysis of BCR-ABL fusion gene transcript revealed the respective fusion gene transcript in 23 out of 25 patients. Out of the 6 patients where cytogenetics failed, 4 patients showed the presence BCR-ABL fusion gene transcript on the RT-PCR analysis. From the study, the following conclusion could be derived. Even though conventional cytogenetics is a gold standard method for the detection of Ph chromosome in patients it could give only a confirmatory diagnosis in 76% of CML patients where as molecular analysis employing RT-PCR for the detection of the bcr-abl fusion gene transcript gave an accurate diagnosis of CML (92%). Key words: Chronic Myeloid Leukemia (CML); BCR-ABL fusion gene; Ph chromosome; cytogenetic analysis; RT-PCR. Authors: Lali Growther* and Razique Punnakkan For Correspondence: E mail :lalijps@yahoo.co.in Tools & Techniques Application Of AFM In Scientific Research Abstract In recent year, Atomic Force Microscope (AFM) has provided a range of new opportunities for viewing, Manipulating and analyzing biomolecules and microbial surfaces in their native environments. And will hopefully allow application to be developed for AFM in Measurement of biological interaction forces, Biotechnology and other bioscience related fields. Hear the sample is realized by interaction of the probe with the sample surface and no imaging beam (light or electron) is involved in the process. The tip of the probe is mounted on the end of a flexible cantilever. 'As the sample is scanned beneath the tip, small forces of interaction with the sample cause the cantilever to deflect, revealing the sample's topography. The most common approach, called an optical lever approach, is to reflect a laser beam off the backside of the cantilever into a 4-segmented photodetector'2. The photodetector generates a measurable signal voltage to make the image. These sophisticated methods are labour- and equipment-intensive, and they are not suitable for examining specimens in aqueous solutions. As well as surface structure, physical properties such as elasticity and surface charge are also important determinants of the functions of cell surfaces. These properties have traditionally been difficult to study in microorganisms. Keywords: Atomic Force Microscope (AFM), Biomolecules, Cantilever, Interaction tip, Piconewton (10-12N). Authors: *R.Rajasekaran, Muthuselvam, M1, C.Arunachalam,2 S. Arunkumar and S. Senthilkumar3 For Correspondence: E-mail: mailtorrajasekarn@ymail.com