Absract Archive
April 2010
Research Methodology Series
Research Ethics in Science and Technology
Introduction
Research in biotechnology/life-sciences has been the backbone towards quality health care and has evolved in terms of complexity. It has advanced rapidly and use of human subjects/samples and animals has now become inevitable in many areas. Ethics has become a serious issue and several key organizations in our country have initiated guidelines, codes of conduct, Standard Operating Practices and Good Laboratory Practices for ethical conduct of research. It is imperative that all organizations, institutes and individuals involved in biological research to be aware of and adhere to the guidelines and protocols as put forth by the competent authorities and committees. In this report, the salient aspects of ethics in using human subjects/samples, animals and stem cells are presented for easy reference and as an update.
Ethics In Research Using Human Subjects/Samples
Key Points
l Policy Statement on Ethical Considerations involved in Research on Human Subjects'- 1980
l Revised version of 'Ethical Guidelines for Biomedical Research on Human Subjects' - 2000
l Revised version of 'Ethical Guidelines for Biomedical Research on Human Participants' - 2006
l Legislation of the 'Ethical Guidelines for Biomedical Research on Human Participants' submitted to Ministry of Health & Family Welfare.
l Draft Ethical Guidelines for Bio-banking in India
l Indian Council of Medical Research - "National Guidelines for Accreditation, Supervision and Regulation of ART Clinics in India"(2005).
Author: M. Ravi
For Correspondence: : E mail : maddalyravi@hotmail.com
Research Article
Bio-active Compounds in Essential Oil and its Effects of
Antimicrobial, Cytotoxic Activity from the Psidium Guajava (L.) LeafAbstract
Leaves of Pisidium guajava L. have been ethnomedically claimed to possess a wide array of biological activities mainly antimicrobial as well as anti-cytotoxic activity. This study was designed to examine the composition of essential oil and antibacterial and antifungal activity. The essential oil (0.13 %) was obtained by hydrodistillation in a modified Clevenger-type apparatus, and their analyses were performed by GC-MS. Identification of the substances was made by comparison of mass spectra and retention indices with literature records. The essential oil was tested against these following ten bacteria (Gram positive and Gram negative) and three fungi. The minimum inhibitory concentration (MIC) of the potent extracts and minimum bactericidal also fungicidal activity were also observed. Furthermore, the essential oil tested against Human Cervical Carcinoma cells (HeLa). Leaves of P. guajava have been ethnomedically claimed to possess a wide array of biological activities including anticancer activity. Subcellular alterations were evaluated by thymidine flouresence viability assay. Cell treated with essential oil showed degeneration of cytoplasmic organelles, reflective reduction, increased shrinkage of the HeLa cell lines and apoptotic characteristics. The results presented here suggested that the essential oil possessed strong antimicrobial as well as probable cytotoxic efficiency, and therefore the essential oil extracted from the fresh leaves of P.guajava (L.) has potential as natural anticancerous and antimicrobial agent in pharmaceutical industry.
Keywords: Psidium guajava, Essential oil, Antibacterial, Antifungal, Gram positive, Gram negative, Cytotoxicity,
HeLa cells.
Authors: Baby Joseph, R. Mini Priya, P.A Mary Helen and S. Sujatha*
For Correspondence: Email: sujatharbs@rediffmail.com
Optimization of Extracellular Peroxidase Production using
Response Surface Methodology from Lentinus SpeciesAbstract
In the present study, four soil samples were collected from various places in the shola forest of Ooty. The soil samples were screened for peroxidase producing fungi. Based upon microscopic and macroscopic features the two fungi were found to be Lentinus spp. and Pleurotus spp. The quantity of peroxidase production was assessed in Czapek dox broth and Lentinus spp. was found to be the better peroxidase producer. The effect of incubation over peroxidase production was studied for a period of 12 days and maximum peroxidase production was found on 10th day. The optimum incubation temperature for maximum peroxidase production was observed at 300C. The influence of pH upon enzyme production was analyzed and the optimum peroxidase production was seen with the pH 6.5. The effect of various carbon sources such as glucose, sucrose, starch, glycerol, sorbitol, fructose, maltose and mannitol was tested at 1% concentration over the peroxidase production was studied and the higher peroxidase production was seen in the medium containing sorbitol. Among the various nitrogen sources peptone induced the maximum peroxidase production. In the several amino acids tested tryptophan was found to enhance the peroxidase production superiorly. Ferrous sulphate was found to be the suitable metal ion since it induced maximum peroxidase production. The concentrations of the sorbitol, peptone and the metallic salt solutions were optimized using statistical approach called response surface methodology and the optimal concentrations was found to be 2.57%, 1.47%, 5.12% for sorbitol, peptone and glycerol, respectively.
Keywords: Peroxidase; Lentinus spp. and Pleurotus spp. Response surface method.
Authors:* P. Sekar, A. Hanitha and G.Gowri
For Correspondence: Email: malarsekarp@gmail.com
Biological Synthesis of Nanocrystalline Silica Particle from TEOS
(Tetra Ethyl Ortho Silicate) mediated by fungi Fusarium oxysporumAbstract
The development of simple and reliable processes for the synthesis of nanosized materials is of great importance in the field of nanotechnology. In this paper, we report on the use of fungi Fusarium oxysporum in the extracellular synthesis of silica nanoparticles. On treating aqueous tetra ethyl ortho silicate (TEOS) solution with fungal mat of Fusarium oxysporum, rapid reduction of the silica ions is observed leading to the formation of highly stable, crystalline silica nanoparticles in solution. The silica nanoparticles were in the range of 10 – 150 nm in dimension. The nanoparticles were examined using UV - Visible absorption spectroscopy, Spectroflourimetry, FTIR spectroscopy, Scanning Electron microscopy and Energy Dispersive X ray spectroscopy analyses. The formation of nanoparticles by this method is simple quick method, requires no toxic chemicals and the nanoparticles are stable for several months. The main conclusion is that the bio-reduction method to produce silica nanoparticles is a good alternative to the chemical methods.
Keywords: Crystalline Nanoparticles, Characterization, MGYP broth, Biosynthesis, Fusarium.
Authors: A. Thangaraja1*, V. Savitha1, A. Kanchana2 and K. Jegatheesan1
For Correspondence: Email: athangaraja@gmail.com
Production of Fusaric Acid From Cassava using Fusarium oxysporum MTCC 1755
Abstract
Fusaric acid (FA) is a mycotoxin produced by the Fusarium species. Fusaric acid is an enzyme inhibitor, dopamine agent and nucleic acid inhibitor. As an orally active agent, it may have potential role in the treatment of head and neck squamous cell cancer. We have used the highest yielding strain, Fusarium oxysporum MTCC 1755. The biosynthesis of Fusaric acid involves condensation reaction polyacetate and aspartic acid units. It can be produced using substrates containing starchy material, such as corn, cassava, rice, potato, wheat etc. We have used cassava, which contains nearly the maximum theoretical concentration of starch on a dry weight basis. It contains almost 30% of starch. We pretreated the cassava by boiling it. We analyzed the sample using titration method and thin layer chromatography. Comparing to the other substrates, we found the maximum yield using boiled cassava. The yield using synthetic starch ranges from 6 to 7 g/L and the yield using unboiled cassava ranges from 7 to 9 g/L and the maximum Fusaric acid production ranges from 9 to 11 g/L using boiled cassava. We also optimized the substrate by varying its concentration. The optimum concentration of boiled cassava was found to be 60 g/L.
Key words: Fusarium oxysporum, Chromatography, Fusaric acid, Cassava.
Authors: B.Bharathiraja1*, M.Jayakumar2, C.Ramaprabu3 and J.Jayamuthunagai4
For Correspondence: E.Mail: btrbio@gmail.com
Electricity Generation from Serratia marcescens Isolated from Aerobic Sludge using Microbial
Fuel Cell Technology and its OptimizationAbstract
The production of energy from wastewater is a high priority for our society given the trends of over population and worldwide energy resource depletion. Aerobic sludge from wastewater treatment plant can be used to produce electricity. MFCs can be operated using pure cultures. Gram negative bacteria like Serratia marcescens can be isolated from it and can be used to produce electricity. It was accomplished with Microbial Fuel Cells (MFCs). A microbial fuel cell (MFC) is a device that converts chemical energy into electricity through the catalytic activities of microorganisms like bacteria. Mutant strain of this bacterium produces higher amount of electricity compared to the wild strain. Although there is great potential of MFCs as an alternative energy source, extensive optimization of this bacterium has been done in this article to exploit the maximum microbial potential from it.
Keywords: Serratia marcescens, Microbial Fuel Cell (MFC), electricity, mutant strain, optimization, salt bridge.
Authors: *K. Ghanapriya1, Sachindri Rana2 and P.T. Kalaichelvan1
For Correspondence: Email Id : gpriyak@gmail.com
Hydrogen Cyanide Mediated Biocontrol Potential of Pseudomonas sp. AMET1055 Isolated from The Rhizosphere of Coastal Sand Dune Vegetation
Abstract
Twenty four hydrogen cyanide (HCN) producing fluorescent pseudomonads (FPs) were isolated from the rhizosphere of sand dune vegetation from Chennai coastal area. All the 24 cyanogenic (HCN producing) FPs were subjected to spectrophotometric assay to relatively quantify the amount of hydrogen cyanide (HCN) produced by them. Five FP strains designated as AMET1039, AMET1041, AMET1042, AMET1055 and AMET1064 produced more amount of HCN in their volatile fraction. In dual bottom plates assay, all these five isolates exhibited maximum mycelial growth inhibition of Rhizoctonia solani MML4001 due to the production of HCN. Among them, AMET1055 was found to be efficient and was identified as Pseudomonas sp. through various biochemical and staining techniques. The microscopical studies have concluded that the inhibitory activity of Pseudomonas sp. AMET1055 against R. solani MML4001 is of fungicidal nature. This study, reports for the first time that the rhizosphere of sand dune vegetation has cyanogenic FPs which can be used as biocontrol agents against soil borne fungal phytopathogens.
Keywords: Hydrogen Cyanide, Cyanogenesis, Fluorescent pseudomonas, Antagonism, Soil borne pathogens.
Authors: M. Jayaprakashvel*, R. Muthezhilan, R. Srinivasan, A. Jaffar Hussain, S. Gobalakrishnan, Jacky Bhagat, C. Kaarthikeyan and R. Muthulakshmi
For Correspondence: Email Id : jayaprakashvel@rediffmail.com
Tutorial Review
In vitro Plant Tissue Culture
Abstract
In vitro plant tissue culture is a practice used to propagate plants under sterile conditions, often to produce clones of a plant. Different techniques in plant tissue culture may offer certain advantages over traditional methods of propagation. It is an important tool in both basic and applied studies as well as in commercial application. It owes its origin to the ideas of the German scientist, Haberlandt, at the begining of the 20th century. The early studies led to root cultures, embryo cultures, and the first true callus/tissue cultures. The period between the 1940s and the 1960s was marked by the development of new techniques and the improvement of those that were already in use. It was the availability of these techniques that led to the application of tissue culture to cell behavior, plant modification and improvement, pathogen-free plants and germplasm storage, clonal propagation, and product formation, starting in the mid-1960s. The 1990s saw continued expansion in the application of the in vitro technologies to an increasing number of plant species. Cell cultures have remained an important tool in the study of basic areas of plant biology and biochemistry and have assumed major significance in studies in molecular biology and agricultural biotechnology.
Keywords: Aseptic technique, In vitro, Plant tissue culture,
Authors: R.Rajasekaran1*, S.Senthil kumar2 and M.Muthuselvam3
For Correspondence: E-mail: mailtorrajasekaran@ymail.com