International Conference on
Latest Trends in Biotechnology and Biodiversity
August 24-26, 2015, Dubai, UAE

Scientific Programme(Day 1 : Aug-24-2015)

Keynote Forum

Zachary Senwo
Alabama A&M University, USA
keynote: Soil Biological and Biochemical Response to Mercury Exposure
Biography:
Dr. Zachary Senwo is Professor and Director of Biological & Environmental Sciences Research programs in the Department of Biological & Environmental Sciences at Alabama A&M University. He received his B.S., from the University of Georgia, M.S., from Alabama A&M University, and Ph.D., from Iowa State University. He is a graduate of the Sparkman Center for Global Health program, University of Alabama, Birmingham and has published over 150 research articles and book chapters impacting research programs globally. He has held various administrative positions at his University and successfully launched scientific partnerships with institutions in Brazil, Costa Rica, China, and Honduras.

Abstract:
Mercury (Hg) although a priority pollutant, persistence in the environment and highly toxic to organisms and humans, has no biological function. We investigated Hg speciation in four soil types spiked with Hg (300 mg kg-1 soil) and its effects on soil microbial respiration and soil enzymes (amidohydrolases and phosphatase). Mercury speciation was investigated under acidic and alkaline conditions while soil microbial respiration and enzymes activities were determined under laboratory settings. Speciation results revealed that the water soluble form was the least, with <1% of the total Hg irrespective of pH condition, while the residual fraction was the most abundant (>80%) in Canisteo, Houston, and Ketona soils under acidic conditions and <35% in Decatur soil. Under alkaline conditions, the residual fraction was ≤70% in Canisteo, Houston, and ketona and ≤29% in Decatur soil. The exchangeable fraction was the second most abundant fraction in the soils used ranging from 3.7 – 50.0% under acid conditions and 16.9 – 52.1% under alkaline conditions indicating that Hg desorption was found to be more favorable under alkaline than acidic conditions. Soil respiration was suppressed by Hg especially at the 100 mg kg-1 concentration level. There was varied responses in amidohydrolases and phosphatase in the presence of Hg. Amidohydrolases were more sensitive to Hg (18 – 90%) than phosphatase (0 – 35%) in all soils. This study demonstrated that the forms in which Hg exist in soils may determine its bioavailability and toxicity. Also microbial respiration and enzyme reactivities are potential bioindicators of heavy metal contaminations of the environment.

Keynote Forum

Harvinder Chawla
G.B. Pant University of Agriculture and Technology, India
keynote: IPR protection in agriculture and biotechnology
Biography:
Prof H.S. Chawla has completed his Ph.D at the age of 23 years from Punjab Agriculture University, Ludhiana. He availed DAAD Fellowship at Institut fur Resistenzgenetik, Gruenbach, Germany and DBT Overseas Associateship at Agriculture Canada, Ottawa. He has 35 years of teaching and research experience in plant biotechnology and IPR areas, published over 100 research publications, books on Plant Biotechnology and IPRs, supervised 16 Ph.D. Presently, he is serving as Head, Genetics & Plant Breeding and CEO, Intellectual Property Management Centre. He served as the Chair in the Asia IPR Conferences in Pakistan, Sri Lanka and Bangla Desh. He facilitated filing of 35 Patents, Designs, Copyrights and registration of plant varieties.

Abstract:
Various international treaties and agreements viz. WTO-TRIPS, CBD, ITPGRFA (Seed Treaty) have been addressing to the issue of plant genetic resources, access and benefit sharing and protection of plant varieties. TRIPs is one of the agreements influencing access to genetic resources and its monopolization. UPOV is one of the most accepted sui-generis system for the protection of new varieties of plants and recognition of plant breeders rights. ITPGRFA provides an international recognition of farmers’ rights. In response to TRIPs agreement provisions, India enacted or amended different laws for protection of genetic resources and biotechnological inventions. India enacted Protection of Plant Varieties and Farmers’ Rights Act, 2001 to protect plant varieties that are new and also traditional farmers’ varieties, which the farmers are conserving and maintaining over the years. Indian PPV&FR Act is unique in the sense that it provides so many rights to farmers which are not even available with UPOV and ITPGRFA. Recent innovations in plant tissue culture, biotechnology, omics and bioinformatics make it necessary to understand the IPR laws. Biotechnology involves application of technology on biological organisms viz. microorganisms, plants and animals and biological material of DNA and RNA but patenting laws of different countries were solely based on non-biological objects and inventions and which are continuously amended. Patenting and protection of microorganisms, plants and animals, cloning, regeneration protocols, promoter, vector sequences and genetic transformation methods, expressed sequence tags (ESTs), molecular markers and scientific disciplines of omics and bioinformatics have been discussed in the context of TRIPs regulations, EPO directive, USPTO and Indian Patent Office guidelines.

Keynote Forum

Fábio de Oliveira
Federal University of Uberlândia, Brazil
keynote: Purification and characterization of BmooAi: a new toxin from Bothrops moojeni snake venom that inhibits platelet aggregation
Biography:
Fábio de Oliveira has completed his Ph.D in 2001 from National University of Brasilia. He was director of Innovation and Transfer of Technology of the Federal University of Uberlândia. Currently is Professor of the Postgraduate Programs in Genetics and Biochemistry and in Cellular and Structural Biology of the Federal University of Uberlândia. Associate Researcher of the National Institute of Science and Technology in Nano-BioPharma and supervisor of postdoctoral, doctoral, masters and undergraduate research students. Has experience in the area of Biochemistry/Biophysics and Biotechnology with emphasis in Isolation and characterization of pharmacologically active principles present in venoms of Brazilian snakes.

Abstract:
In this work, I describe the purification/characterization of BmooAi, a new toxin from Bothrops moojeni that inhibits platelet aggregation. The purification of BmooAi was carried out through three chromatographic steps (ion-exchange on a DEAE-Sephacel column, molecular exclusion on a Sephadex G-75 column and reverse-phase HPLC chromatography on a C2/C18 column). BmooAi was homogeneous by SDS-PAGE and shown to be a single-chain protein of 15,000 Da. BmooAI was analysed by MALDI-TOF Spectrometry and revealed two major components with molecular masses 7824.4 and 7409.2 as well as a trace of protein with a molecular mass of 15,237.4 Da. Sequencing of BmooAi by Edman degradation showed two amino acid sequences: IRDFDPLTNAPENTA and ETEEGAEEGTQ, which revealed no homology to any known toxin from snake venom. BmooAi showed a rather specific inhibitory effect on platelet aggregation induced by collagen, adenosine diphosphate or epinephrine in human platelet-rich plasma in a dose-dependent manner; whereas it had little or no effect on platelet aggregation induced by ristocetin. The effect on platelet aggregation induced by BmooAi remained active even when heated to 100°C. BmooAi could be of medical interest as a new tool for the development of novel therapeutic agents for the prevention and treatment of thrombotic disorders.

Keynote Forum

Benoit Schoefs
University of Le Mans, France
keynote: Microalga Milking: a Review and New Approaches
Biography:
Benoît Schoefs graduated from the U Liège (ULg). He became Professor in Plant Physiology at the U Dijon. Recently, he joined the U Le Mans (France). He has published more than 66 papers in international journals and edited books, monographies and special issues. He is a member of the editorial board of International Journal of Plant Biology, Communicative and Integrative Biology and Acta Botanica Gallica. He won the Prize ‘D. Clos’ from the Toulouse Academy of Sciences and the annual scientific competition of the Belgian Academy of Sciences. His research is focused on the responses of photosynthetic organisms, including algae, to stresses. He heads the research team MicroMar.

Abstract:
The rise of human populations, potential climatic changes, and the growth of cities contribute to the depletion of natural resources and increasing their cost. To overcome difficulties in supplying populations and reducing the resource cost, a search for alternative energy, pharmaceutical and nanotechnology sources has started. Among them, microalgae are very promising because they use carbon dioxide to produce biomass and/or valuable compounds. Once produced, the biomass is harvested and processed through a dedicated downstream programme. Drying, grinding and extraction steps are the most common steps of such programmes and are destructive to the microalgal biomass that then needs to be renewed. In addition, extraction and purification steps generate organic wastes. Altogether, it is urgent to develop alternatives, more environmental friendly downstream processes to process algal biomass. Among the possibilities, the concept of milking is especially interesting because it specifies that the extraction should not kill the cells. Therefore, it does not require the need of culturing and regrowing the algae. In this contribution, the development of milking of microalgae is discussed. The main grounds for thought identified are (a) development of alternative methods to extract and harvest high added value compounds, (b) design of photobioreactors, (c) biodiversity and (d) stress physiology. They are discussed separately and illustrated with original results.

Cancer and Genomics Research
Genetic Engineering and rDNA Technology
Biotechnology in Agriculture

Session Introduction

Ajay Kumar Garg
UAE University,United Arab Emirates
Title: Improving salinity and drought stress tolerance in transgenic plants by stress-inducible expression of novel genes
Biography:
Dr. Ajay Garg obtained his Ph.D. degree in Plant Sciences from the University of Hyderabad (1996). He began his career as a Rockefeller Foundation Post-doctoral Fellow at Cornell University in 1996, and became the Rice Project Leader in the Department of Molecular Biology and Genetics, and the Visiting Scientist in Robert Holley Center for Agriculture and Health (USDA-ARS) and the Chief Scientific Officer for Ray Wu Agritech at Cornell Center for Technology, Enterprise and Commercialization. He spent about 15 years at Cornell University and has made outstanding contributions and published several important research articles on the production of useful transgenic cereal plants for abiotic stress tolerance and is an Inventor of several US patent or international patent applications. In 2011, he joined UAE University as an Associate Professor of Plant Biotechnology and is working on Date Palm genetic engineering and biotechnology research.

Abstract:
Genetic engineering of plants with an agronomically useful gene(s) is a powerful tool for producing valuable transgenic plants to feed the growing world population. As we know, salinity and drought stress results in substantial yield losses and pose serious threats to agricultural production and food security worldwide. In addition, about 30% of the land in the world contains levels of salt too high for optimal food production. Therefore, generating abiotic stress tolerant transgenic plants could be beneficial in an era of global climate change and fresh water crisis. Acclimation of plants to abiotic stresses such as drought, salinity, or heat is mediated by a complex network of regulatory genes and other transcriptional factors that control multiple defense enzymes, proteins, and pathways. Here, we will report the results of stress-inducible or tissue specific expression of more than ten different candidate genes or transcriptional factors on abiotic stress tolerance in cereal plants. As compared with non-transgenic plants, many independent transgenic plants exhibited sustained plant growth and less photo-oxidative damage under environmental stress conditions. We will also try to compare our results with previously published work in many transgenic monocot and dicot plants.

Arokiaraj Pappusamy
Manipal International University, Malaysia
Title: Biopharming in Hevea brasilienis
Biography:
Arokiaraj Pappusamy has completed his Ph.D from King’s College, University of London and postdoctoral studies from University of Hertfordshire School of Life Sciences United Kingdom. He is a British Chevening Scholar and a recipient of the Royal Society Malaysian Fellowship Award. He has a patent for the production of proteins in plant fluids. Currently, he is the Head Department of Biotechnology, School of Science and Engineering at Manipal International University Malaysia, under Manipal Global Education. He has published more than 45 papers in reputed journals and a member of Society for In Vitro Biology, USA.

Abstract:
As in most crop plants, the objective of genetic transformation is to develop genetically modified plants with desirable agronomic traits. More than that, researchers have another major research target and that is for the production of valuable proteins. It is here that we seek to turn the transgenic rubber tree (Hevea brasiliensis) into a factory for the production of valuable proteins such as pharmaceuticals. The rubber tree is a unique plant in that its crop harvest, through latex tapping, is non-destructive in nature. This confers upon the tree an advantage not found among other crop plants that are engineered to produce recombinant proteins. Here, the production of the target recombinant protein in the tapped latex is continual. In this connection, previous research findings in producing β-glucuronidase (a bacterial enzyme-routinely used for the enzymatic hydrolysis of glucuronides), a recombinant antibody and human serum albumin in the latex of transgenic rubber plants were very encouraging. More recently, a gene encoding human atrial natriuretic factor (HANF), a peptide hormone that is involved in regulating cardiac bold pressure was inserted into rubber cells and the results of this experiment shall be presented. Routinely, the CaMV 35S promoter is employed to express transgenes in Hevea, which results in synthesis of recombinant proteins at low levels. One way to increase the levels of recombinant proteins is by manipulating the gene constructs employed in genetic transformation. Here, we employed a latex-specific promoter (hevein promoter) to overexpress β-glucuronidase in the latex of rubber plants. The transgenic Hevea system has shown itself to be a promising technology for ‘molecular pharming’ in the future.

Muhammad Ilyas
Abdul Wali Khan University, Pakistan
Title: Whole genome sequencing and analysis of a Pakistani human genome for clinical and evolutionary inference
Biography:
Muhammad Ilyas is Assistant Professor at Abdul Wali Khan University Pakistan. He is using some innovative computational approaches to detect ethnic and even tribal-specific mutations that may be the key to rare and common diseases with higher prevalence in the population under study. He worked on genome-wide data analysis of human populations in Pakistan and their genetic relationship with other human populations in order to evaluate the genetic background of these groups. His aim is to provide insights into the processes that have modeled the extant genetic diversity in these populations (such as migrations, bottlenecks and admixture) as well as the consequences of the diversity observed, including their relation to disease susceptibility. He has the experience to work on different positions at the CEMB (Pakistan), KOBIC (Korea), PGI (Korea) and Harvard (USA). Throughout his career Ilyas has won numerous awards and scholarships including the HEC international fellowship, Sandwich doctorial scholarships, Presidential Award, Best Young Scientist Award, Best Research Paper Award (HEC) and was awarded as Productive Scientist in 2012 by PCST. He founded a Regional Student Group in Pakistan (RSG Pakistan) affiliated with ISCB, in the year 2010, for developing computational biology in Pakistan.

Abstract:
Pakistan covers a key geographic area in Asia and played an important role in human history, being both part of the Indus River region that acted as one of the cradles of civilization and as a link between Western Eurasia and Eastern Asia. Here we present a whole-genome sequence and analyses of a northwestern Pakistani individual. A total of 3.8 million single nucleotide variations (SNVs) and 0.5 million small indels were identified (Novel: 129,441 and non-synonymous SNVs: 10,315 in 5,344 genes). SNVs were annotated for health consequences and high risk diseases, as well as possible influences on drug efficacy. We confirmed that the Pakistani genome presented here is representative of this ethnic Pakhtun group. Finally, we reconstruct the demographic history by PSMC, which highlights a recent increase in effective population size compatible with admixture between European and Asian lineages expected in this geographic region. It is a useful resource to understand genetic variation and human migration across the whole Asian continent.

Maria Maqsood
Forman Christian College (A Chartered University), Lahore, Pakistan
Title: Molecular characterization of some bacterial isolates from sugarcane and their effect on growth of corn plants
Biography:
Maria Maqsood has completed her M.Phil Biotechnology from Forman Christian College (A Chartered University) where she was awarded an academic excellence award by DuPont Pioneer to work in agricultural biotechnology. She is currently a lecturer at Bahauddin Zakria University, Lahore, Pakistan and has two international publications. Her research interests include plant microbe interaction and the use of phylogenetic markers to screen plant growth promoting rhizobacteria for sustainable development.

Abstract:
Characterization of microbial diversity is an important step towards identification of bacteria that could be used for their beneficial qualities such as Plant Growth Promoting Rhizobacteria. The current study characterized bacterial isolates from sugarcane based on two phylogenetic markers, cpn60 and 16S rRNA gene. Eight bacterial isolates were assessed for their impact on the growth of corn plants. Out of the eighteen isolates that were characterized through the cpn60 marker and phylogenetically compared, the most dominant genus observed was Bacillus and Enterobacter. Bacteria with PGPR traits were also identified including Bacillus cereus, Enterobacter cloacae, Citrobacter freundii, Klebsiella pneumoniae, Burkholderia cepacia, Aeromonas hydrophila, Bacillus thuringiensis and Psuedomonas putida. These bacteria have been previously reported for their positive effect on the growth of various plants. Out of eight bacterial isolates identified through the 16S rRNA marker, two of the bacteria were identified as Enterobacter cloacae subsp. dissolvens. Two bacterial isolates each were identified belonging to the Bacillus and Kosakonia genus. Two bacterial isolates showed similar identification through the cpn60 and 16S rRNA marker. All eight isolates used in the plant experiment showed positive significance when compared with uninoculated plants. Isolates PB-CS2 and 77NST-W2 showed highest values for all parameters compared. 20-30 percent increase was observed in root length, 15-20% in shoot length, 30-40% in root area, 20-35% in shoot area, 15-30% in root fresh weight 13-30% in shoot fresh weight, 20-40% in root dry weight and 15-40% in shoot dry weight. Isolates identified in this study are important from the perspective of being used as bioinoculants as well as being generally important bacteria.

Anjana Jajoo
Devi Ahilya University, India
Title: Chlorophyll a Fluorescence: a quick tool to monitor plant’s health
Biography:
Anjana Jajoo is an Associate Professor in School of Life Science of Devi Ahilya University (DAU), Indore, India. She obtained her doctorate degree in Life Science from DAU (1993).She has authored/co-authored more than 60 research papers, several book chapters, has guided 15 doctorate students. She has availed several fellowships from national and international funding agencies in order to carry out research work in several renowned laboratories all over the world. At the same time, she has completed several national and international collaborative projects. Her research interests include abiotic stress responses in photosynthetic machinery, particularly Photosystem II.

Abstract:
Crop yield is directly related to its photosynthetic efficiency. Photosynthesis is also one of those processes which get significantly affected by biotic or abiotic stress. The physiological state of several photosystem II (PSII) components, electron transport chain components, and the cooperation of light-dependent and light-independent biochemical reactions can beevaluated by analysis of chlorophyll fluorescence (ChlF) induction curves. ChlF potentially has extensive applications because chlorophyll a is present in all organisms capable of oxygenic photosynthesis (Embryophyta, algae, lichens, and cyanobacteria). Research techniques based on ChlF measurements are being effectively used in a range of research areas such as plant physiology and plant protection , bioenergetics, agriculture, horticulture,forestry , plant biology,biotechnology, plant breeding, ecology, warehousing of vegetables and fruits, and food technology and processing. ChlF measurements are also valuable for estimating the quality of fruits, vegetables, and flowers and for defining the best time to sell them. It is a popular technique because it is very quick, non-invasive, non-destructive and reliable method. Measurement methods based on ChlF are among the most important instruments currently used in plant breeding programs due to their ability to indicate the physiological state of plants, providing insights into plant growth and yield under naturally occurring environmental stress conditions. Illumination of a dark-adapted photosynthetic sample allows a polyphasic chlorophyll fluorescence induction curve (O-J-I-P-transient). The shape of the curve changes under different physiological conditions. The curve’s characteristic course gives substantial information about the structure and function of the photosynthetic apparatus.

Bushra Hafeez Kiani
International Islamic University, Pakistan
Title: Artemisinin production in Artemisia annua and Artemisia dubia following transformation with the rol ABC genes and elucidation of the sites of its synthesis.
Biography:


Abstract:
The rol ABC genes have been shown to enhance production of secondary metabolites in plants, possibly through stimulation of the defense pathway. This report examines the effect of transformation of A. annua and A. dubia with the rol ABC genes expressing in A. tumefaciens and A. rhizogenes. The artemisinin content, trichome density and expression of key genes in the biosynthetic pathway of artemisinin were measured. Artemisinin content was significantly increased in transformed material of both Artemisia species when compared to un-transformed plants. The artemisinin content within leaves of transformed lines was increased by a factor of ten, indicating that the plant is capable of synthesizing much higher amounts than has been achieved so far through traditional breeding. Expression of all artemisinin biosynthesis genes was significantly increased, although variation between the genes was observed. Cytochrome P450 (CYP71AV1) and aldehyde dehydrogenase 1 (ALDH1) expression levels were higher than that of amorpha-4, 11 diene synthase (ADS). Levels of the trichome development and sesquiterpenoid biosynthetic gene (TFAR1) expression were also increased in all transgenic lines. Trichome density was also significantly increased in the leaves of transformed plants, but no trichomes were found in control roots or transformed roots. The detection of significantly raised levels of expression of the genes involved in artemisinin biosynthesis in transformed roots correlated with the production of significant amounts of artemisinin in these tissues. This suggests that synthesis is occurring in tissues other than the trichomes which contradict previous theories. This elucidation will help to increase production to meet the increasing demand of artemisinin because of its pharmacological importance.

Laid Benderradji
Mentouri University, Algeria
Title: Effect of some abiotic stresses and growth regulators on somatic embryogenesis of mature and immature bread and durum wheat embryos (Triticum Sp.)
Biography:


Abstract:
Mature embryos of (Triticum sp.) cultivars, Oued Zenati (OZ), Djenah khetifa (DjK) and Waha (WH) as a durum wheat (Triticum durum Desf.) and Aïn Abid (AA), Mexipak (MX) and Mahon Demias (MD), as a bread wheat (Triticum aestivum L.), were studied in vitro under thermal and saline stress within 2,4 dichlorophenoxyacetic acid (2,4 D.) as an auxin. The experiment was carried in genetics, biochemistry and plant biotechnology laboratory of Constantine university-Algeria. Callus induction is obtained from all variaties embryos placed on Murashigue and skoog (MS) medium in addition to 2.4 D at 5.5mM for 4 weeks, then callus showed hight developpment were separately placed in (MS) added of various NaCl concentrations (0, 85, 171 and 255mM), and different thermal degrees (25, 30, 35 and 40°C), during 3 hours. After 3 weeks in darkness, the uniform explants were transferred on (MS) added of BAP at 22.19mM and NAA at 10.74mM for foliar induction (caulogenesis) and 4 weeks afterwards on half strength (MS/2) added of Kinetin at 4.92mM and NAA at 10.74mM for radicular induction (rhizogenesis). Results show significant a genotype effect for somatic embryogenesis capacity and plantlets regeneration under abiotic stresses.

Ariful Islam
EXIM Bank Agricultural University, BANGLADESH
Title: Estimation of General Combining Ability (GCA) Effects in Morpho Reproductive Traits of Rice (Oryza sativa L.)
Biography:
Dr. Md. Ariful Islam has completed his Ph.D at the age of 30 years from department of Genetics and Plant Breeding, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Gazipur, Bangladesh. He is assistant professor at department of Genetics and Plant Breeding, EXIM Bank Agricultural University, Bangladesh. Previously he served as Senior Plant Breeder and head, hybrid rice research and production GETCO agro vision limited, Bangladesh from 2009-2014. He has published more than 12 papers and 15 articles in national and international proceedings in reputed journals. He has interest on conventional plant breeding, molecular genetics, cyto-genetics to comment on gene transfer, recombinant DNA transfer and genetic nature of developed and introduced hybrids, clones and inbreds.

Abstract:
Combining ability effects of newly developed CMS and restorer lines were estimated at the experimental farm, of department of Genetics and Plant Breeding, Bangabandhu Sheikh Mujibur Rahman Agricultural University, Salna, Gazipur during Rabi 2011 following RCBD design in three replications. GCA variances of five CMS lines and sixteen newly developed R-lines were estimated through line × tester analysis. Out of sixteen restorer lines eight R-lines showed significant negative effects for days to 1st flowering, seven showed significant negative effects for days to 80% flowering and five showed significant negative effects for days to maturity. Among 16 restorer lines, only four showed significant nigative gca effects for all these three traits. Three pollen parents and one CMS line, showed significant positive gca effects for pollen fertility while six pollen parents showed significant positive effects for spikelets fertility but two pollen parents showed significant positive gca effects for both panicle and stigma exertion rate. The estimated of gca effects of parents indicated that RG-BU 08-001R, RG-BU 08-002R, RG-BU 08-0046R, RG-BU 08-0057R, RG-BU 08-0097R, BRRI 1A and IR62829A contributed highly significant negative effects for plant height which were responsible for dwarfing character. More panicle bearing tillers per plant is believed to be closely associated with high grain yield per plant resulting high productivity. Nine parents contributed significant positive alleles for higher effective tillers per plant. And finally significant positive gca effects was found in seven pollen parents for grain yield. Perse performances revealed that these seven pollen parents as well as two CMS lines were the best general combiner due to highly significant positive gca effects.

Bushra Ijaz
University of Punjab, Pakistan
Title: Combinatorial RNAi Approach against HCV
Biography:
Dr. Bushra Ijaz did her Ph.D in Molecular Biology from Centre of Excellence in molecular Biology, University of the Punjab, Lahore, Pakistan. She has been serving in the same Centre since 2006 after her M.Phil first as research officer and now as lecturer. She has published 25 research articles in reputed journals. Ms. Ijaz has experience working in molecular biology with particular focus on applied and functional Genomics.

Abstract:
HCV infection has affected a large number of populations worldwide. Presently available therapies are expensive, prolonged and associated with significant adverse effect. So, there is a clear need for the development of additional agents that act through alternate mechanisms. RNAi appears to be an effective nucleic acid-based gene silencing tool to target highly conserved or functionally important regions within the HCV genome. A number of strategies have been employed for in vivo delivery of siRNA, but now a day, intracellular production of siRNA through plasmids containing shRNA expression units is one of the promising methodologies in use. One advantage of using vector based strategy is to produce more than one siRNA, targeting more than one gene. Utilizing this characteristic of shRNA delivery mechanism, we have employed combinatorial siRNA strategy expressing two siRNA under single promoter against HCV. We targeted Core and 5’UTR genes of HCV, because both of these genes play very crucial role in HCV life cycle and are most conserved comparing to other genes. Moreover, to evaluate the efficacy of shRNA against the whole virus, the cell culture tested shRNA was checked for inhibition of HCV replication in Huh-7 cells infected with HCV 3a serum. Results showed drastic reduction in (80 to 90% inhibition) in viral titer. This clearly demonstrates that the RNAi-mediated silencing of the HCV genes resulting from mRNA degradation can be an effective siRNA based therapeutic opportunity against HCV.

Abdul Hannan
Ghazi University, Pakistan
Title: Production of Dual Action Mycopathocide by Novel species of Trichoderma harzianum Isolated from Naturally Diseased Suppressed Soil
Biography:


Abstract:
Mycofungicide and biofertilizers are considered very important in green agriculture. Current research is an effort to exploit actual potential of local strains of Trichoderma harzianum to manage soil borne plant pathogens (SBPPs). The main thrust of project is to introduce a biocontrol product for vegetable growers and specifically for kitchen gardeners. During 2014, a study was conducted and diversity of Trichoderma genus was determined in Faisalabad Division, Punjab, Pakistan. The sampling sites were selected from cultivated vegetable fields. A total of 388 soil samples were collected along with fact sheet for each sample. All the information regarding disease incidence and pesticide application history was recorded for each sample. After initial scrutiny 284 samples were processed further and four species of Trichoderma (Trichoderma harzianum, T. hamatum, pseudokoningii, and T. konigii) were isolated and identified following guidelines from Rifai, 1969 and Bissett, 1991.The frequency of T. harzianum isolates was maximum (96.2%) with average of 3.2X103cfu/g from examined samples. Therefore, only strains T. harzianum were further tested for antagonistic potential against target SBPPs (Fusarium oxysporum, Alternaria Alternata, Rhizoctonia solani and Macrophomina phaseolina). The antagonistic potential was determined through Dual culture plate method against all SBPPs. The data revealed that the strains of T. harzianum isolated from naturally diseased suppressed areas showed higher percentage of reduction in colony diameter over control (68.8 to 78.2%) during dual culture test against SBPPs. Moreover, the results showed a detrimental interaction between population of SBPPs and antagonistic potential of biocontrol agent with positive correlation. The data analysis depicted that the disease incidence increased by 0.78% if one unit increment in frequency of T. harzianum was observed. The studies were further extended to characterization and cultivation of T. harzianum on different substrates. Additionally, the strains with higher pathocidal prospective were characterized for maximum yield of conidia on cheaper substrates under solid-state fermentation. All substrates showed dissimilar response but conidial count was maximal in rice bran supplemented with chickpea flour (2.5 x 108cfu/g) after 18 days of incubation at 27ºC. The characterized strains were tested in the field where reduction in disease incidence over control was observed (52.4 to 68.0%) over control. These findings explored a possibility of cost effective technology for production of T. harzianum at pilot scale. The current studies are still in progress, where different formulation of substrates are being tested and large scale production of selected and characterized strains is under the process of optimization. Finally a standard bioassay will be conducted for quality control and product will be patented and registered.

Saima Rehman
Government College University, Pakistan
Title: HPLC assay of caffeine and paraxanthine for CYP1A2 phenotype assessment
Biography:
Saima Rehman has completed his Ph.D at the age of 23 years from University of Agriculture. Recently she is working as Controller of Examination in Govt. College university, Faisalabad. During her Ph.D. research in USA, she got expertise in genetic tools and accessent of risk factors.

Abstract:
Caffeine, not cancer causing agent or not decreases the risk of cancer but used just to note the activity of Cytochrome P-450 1A2 by converting into its metabolite i.e., paraxanthine. The purpose of present study was to determine the caffeine and its metabolite phenotypes and their relation to cancer risk in healthy female volunteers of local population in Pakistan. The average value of metabolic ratio was found to be 1.182995 ± 0.21137. BMI of all volunteers were found to be 19.93Kg/m2. Retention time was 15 and 37mins for 17X and 137X respectively. The linearity of calibration curve of 137X and 17X were covered 0 – 12 µg/ml (R2 = 0.994). A significant positive correlation was observed between metabolic ratio and cancer risk factors. We could conclude that all the volunteers are fast metabolizers having greater risk of cancerous diseases.

Biotechnology in Health Care
Food and Bioprocess Technology

Session Introduction

Jian-Yong Wu
The Hong Kong Polytechnic University, China
Title: Ultrasonic disruption for efficient extraction of polysaccharides and proteins from viscous fermentation broth of a medicinal fungus
Biography:
Dr. Jian-Yong Wu completed his PhD and postdoctoral studies on Biochemical Engineering in 1994 at Queen’s University in Kingston, Canada. He is now an Associate Professor of Food and Bioprocess Technology at The Hong Kong Polytechnic University. He has published more than 140 peer-reviewed papers in reputed scientific journals, received over 2400 ISI citations and an H-index 30. He has been serving as an editorial board member of Biotechnology Applied Biochemistry. His current areas of research interest include: bioprocesses with medicinal fungi and plant tissue cultures; food and medicinal chemistry and functions with polysaccharides from medicinal fungi and plants; ultrasound-assisted processes for natural and medicinal products.

Abstract:
Introduction and Objectives: Polysaccharides including PS-protein complexes (PSPs) are major bioactive constituents of edible/medicinal fungi with notable antitumor, immunomodulatory, antioxidant activities and other health benefits (Wasser 2002; Stachowiak and Regula 2012). Liquid or submerged fermentation is most commonly applied for mass production of mycelial biomass and useful metabolites. In liquid fermentation, most of the bioactive metabolites synthesized by the fungi are retained in the mycelial biomass and some are released into the liquid medium, such as exopolysaccharides (EPS) produced by some medicinal fungi. Cordyceps (Cordyceps sinensis), generally known as the Chinese caterpillar fungus, is a rare and precious medicinal fungus which has been used as a favorable tonic for hundreds of years in China (Chen et al. 2013; Zhu et al. 1998). Because of the very limited supply and high value of natural Cordyceps organisms, mycelial fermentation has become the most viable process for mass production of fungal biomass and polysaccharides. Cs-HK1 is a fungus isolated from a natural Cordyceps fruiting body, and Cs-HK1 mycelial culture has been established for production of medicinal materials through liquid fermentation (Leung et al. 2006; Wu et al. 2014). However, the fermentation broth containing filamentous fungal mycelia and high-molecular weight EPS is highly viscous and very difficult to separate. In this study, high-intensity ultrasound (US) was applied as a simple and effective means to facilitate the extraction of intracellular and extracellular polysaccharide-protein complexes (PSPs) from the viscous Cs-HK1 mycelial fermentation broth.

Pradman K. Qasba
NCI, NIH, USA
Title: Applications of Glycosyltransferases in the Site-specific Conjugation of Biomolecules and Development of a Targeted Drug Delivery System and Contrast Agents for MRI
Biography:


Abstract:
The structural information on glycosyltransferases from our laboratory has revealed that the sugar-donor specificity of these enzymes can be broadened to include modified sugars with a chemical handle that can be utilized for conjugation chemistry. Substitution of Tyr289 to Leu in the catalytic pocket of bovine -1,4-galactosyltransferase generates a novel glycosyltransferase that can transfer not only Gal but also GalNAc or a C2-modified galactose that has a chemical handle, from the corresponding UDP-derivatives, to the non-reducing end GlcNAc residue of a glycoconjugate. Similarly, the wild-type polypeptide-N-acetyl-galactosaminyltransferase, which naturally transfers GalNAc from UDP-GalNAc, can also transfer C2-modified galactose with a chemical handle from its UDP-derivative to the Ser/Thr residue of a polypeptide acceptor substrate that is tagged as a fusion peptide to a non-glycoprotein. Also many GlcNAc-transferring enzymes, both mutant and wild-type, can also transfer the GlcNAc analog C2-keto-Glc molecule from UDP-C2-keto-Glc to their respective acceptor substrates, suggesting that the N-acetyl-binding cavity for the donor sugar substrate in the glycosyltransferases can generally also accommodate a chemical handle in the N-acetyl-binding cavity. The availability of the modified sugar moiety thus makes it possible to link cargo molecules at specific sites. The cargo may be comprised of, for example, biotin or fluorescent tags for detection, imaging agents for magnetic resonance imaging (MRI), or cytotoxic drugs for cancer therapy. The potential of wild-type and mutant glycosyltransferases to produce glycoconjugates carrying sugar moieties with chemical handle makes it possible to conjugate cytotoxic drugs, cytokines, or toxins for antibody-based cancer therapy and the development of a targeted drug-delivery system or fluorophores for ELISA-based assays, radionuclides for imaging and immunotherapy applications, lipids for the assembly of immunoliposomes.

Fabio Apone
Arterra Bioscience /Vitalab srl, Italy
Title: Plant tissue cultures are valuable sources of bioactive ingredients for cosmetic and nutraceutical applications.
Biography:
Dr. Fabio Apone graduated in Biology in 1994 and obtained his PhD title in Protistology (Biology of Unicellular Organisms) in 1998 at the University of Pisa, Italy. He worked for 3 years as Postdoc Researcher in Italy and at the University of California, San Diego, focusing on Cell and Molecular Biology. Later he was a research scientist at Arena Pharmaceuticals Inc, a Biotech Company in California, studying plant receptors for the discovery of novel agrochemicals. He is currently the Scientific Director and Project Coordinator at Arterra Bioscience and VitaLab, companies focused on the development and commercialization of cosmetic active ingredients.

Abstract:
Human health care research is constantly looking for innovative plant ingredients, which must be guaranteed for quality and safety to the final consumers. Unfortunately, many plant derived products can be used limitedly because several plants of interest may contain toxic compounds, can be subjected to diseases and differ in the content of metabolites seasonally and from harvest to harvest. By using biotechnological approaches, plant cells and tissues can be easily cultivated in sterile conditions, totally independently of geographical and climatic factors, thus represent a very relevant system for the production of valuable metabolites. Moreover, differently from plants grown in the field, ingredients obtained from plant tissue cultures can be always guaranteed for safety, quality and sustainability, main priorities for health care product consumers. Exploiting the natural biodiversity and versatility of the plants, we successfully employed plant suspension and hairy root cultures as multiple sources of extracts having different chemical nature, and a wide range of therapeutic activities. By using human skin cultures and intestine cell models, we evaluated the efficacy of the extracts as cosmetic or nutraceutical ingredients. The capacity of the compounds present in the extracts to protect the cells against oxidative, UV, inflammatory or biotic stress was deeply investigated in order to find the best ingredient or combination aimed to a specific and desired application in the health care industry.

Vijay Prabha
Panjab University, India
Title: Efficacy of antisperm receptor monoclonal antibodies as vaginal contraceptive
Biography:
Dr. (Mrs) Vijay Prabha, is working as Professor in the Department of Microbiology, Panjab University, Chandigarh, India. She has 25 years of teaching and 30 years of research work experience. She has guided number of M.Sc. and Ph.D students. She has about 60 publications in national and international journals. She is life member of Association of Microbiologists of India and Panjab University Research Journal of Science. She is on the reviewer panel of various international and national journals. She is also member of National Editorial Advisory Board of Indian Journal of Microbiology Research and Journal of Microbiology and Related Research.

Abstract:
Objective(s): In an earlier work performed in our laboratory, we have been able to isolate a sperm receptor (SR; ~125kDa) from human spermatozoa exploiting bacteria sperm interactions at the receptor ligand level, playing critical role in fertilization. Polyclonal antibodies generated against SR resulted in failure of conception in female BALB/c mice when administered intravaginally. The generation of monoclonal antibodies against SR, selected on the basis of its potency, specificity and safety might result in generation of an improved vaginal barrier, hence in the present work, an attempt was made to generate anti-SR monoclonal antibodies and their impact was checked on fertility outcome in NZW rabbits. Study Design: Monoclonal antibodies against the purified SR were generated using mouse myeloma cell line (Sp/2OAg.14) in BALB/c mice. The concentration of IgG in hybridoma supernatant was estimated using Easy-titre® mouse IgG Assay kit and the specificity of the hybrids so obtained was checked on the basis of ELISA and presence of spermagglutinating activity in vitro. The isotyping of IgG was done using immunodiffusion assay. The supernatant from the wells of positive clone/s was administered intravaginally in female NZW rabbit before mating. Control rabbits received supernatant from wells containing only mouse myeloma cell line. Further, the effect of consecutive intravaginal applications (14 days) of anti-SR mAbs was checked on reproductive organs viz. vagina, uterus and ovary. Results: From a panel of mAbs raised against SR, one clone designated as G11 could induce sperm agglutination at a concentration of 85µg within 15min of incubation of human spermatozoa and showed highest specificity towards SR in vitro. The subclass of immunoglobulin was found to be IgG2b. Intravaginal application of anti-SR mAbs (G11; 100µg) resulted in failure of conception in NZW rabbits. Further, absence of any infiltration of polymorphonuclear leukocytes or inflammatory reactions was observed after 14 consecutive intravaginal administrations on any of the reproductive organs tested. Conclusion(s): From these results, it could be concluded that the isolated SR may be potential candidate antigen for the development of vaginal contraceptive as it could induce infertility following passive immunization.

Sadeq Shabani
Biotechnology Research center of Pasteur Institute of Iran, Iran
Title: Designing and evaluating internal control for qualitative and quantitative HIV detection kits
Biography:
Sadeq Shabani, graduated with Master of Molecular Genetics in Pasteur Institute of Iran. Along with several research works in Biotechnology Research Center of Pasteur Institute of Iran and working R&D Department of Roya Bio Gene Company, genetics and Biotechnology Company, as a head of Department.

Abstract:
Viral infections are main diseases in the world, and acquired immunodeficiency syndrome (AIDS) (the end phase of human immunodeficiency virus [HIV] infection) has been one of the most dreaded infectious diseases causing mortality in individuals. There are a lot of cultural methods for detection of viral infections, but they are time-consuming. Recently, old methods have been substituted by Molecular diagnostics methods such as PCR and Real Time. However, there are many problem associate with molecular methods such as lack of standard. In this research, the problem was solved by designing competitive Internal Control(IC). In this research, for designing competitive internal control, first two primers were designed to detect HIV then PCR Amplification was performed to amplify expect fragment. After that, composite primer (HIV- IC) was designed for adding 20 nucleotide of leishmania gene to both terminal of the PCR product. Finally amplified fragment by composite primer TA cloned and used as IC. We also evaluated minimum of 5 copy numbers IC in one reaction that detect by Real time PCR and 500 copy number by simple PCR assay. We hope these efforts help to better detection HIV disease and decrease rate of error-born.

Hajer Taleb
Cardiff Metropolitan University, UK
Title: The bacteriostatic and bactericidal activity of date syrup and its potential mode of action
Biography:


Abstract:
Date Syrup has been produced for many centuries as an incidental product of date fruit storage, when bagged dates are stored together over several months. Date syrup has traditionally been used in the Middle East to treat a wide range of diseases and disorders such as wound healing, hypertension, oedema and gastrointestinal disease. In some cases these ailments can be attributed or are strongly associated with a variety of bacterial infections and inflammation. Plant-derived products such as date syrup can inhibit bacteria through numerous different mechanisms, which may be attributed to the presence of bioactive compounds including plant-derived phenolic molecules. Many such products derive their inherent antimicrobial activity from the presence of polyphenols which cause oxidative damage. Here we have demonstrated that date syrup, and date syrup polyphenols, the most abundant bioactive constituent of date syrup are bacteriostatic and bactericidal to a range of Gram positive and Gram negative disease causing pathogens such as Escherichia coli and Staphylococcus aureus. We have further shown that the isolated polyphenols independently suppress the growth of bacteria and have observed that date syrup behaves as a prooxidant to produce hydrogen peroxide that mediates bacterial growth inhibition. This effect is in conjunction with pH and osmolarity; under weakly acidic conditions date syrup demonstrated anti-oxidative activity by reducing hydrogen peroxide, whereas under neutral (optimum bacterial growth conditions) and weakly alkaline conditions date syrup demonstrated prooxidant activity that inhibited the growth of E. coli and S. aureus. The high sugar content naturally present in date syrup did not significantly contribute to this effect. These findings highlight that the inhibition of bacteria is mediated through hydrogen peroxide, and is influenced by pH in inducing oxidative stress in bacteria.