Day 2 :
Keynote Forum
Sandeep Kumar Vashist
University of Freiburg Germany
Keynote: Smartphone-based colorimetric readers for in vitro diagnostics and mobile healthcare
Time : 09:30-10:00
Biography:
Sandeep K. Vashist is the Head of Immunodiagnostics at Hahn-Schickard-Gesellschaft für angewandte Forschung e.V. (HSG-IMIT) and Laboratory for MEMS Applications, Department of Microsystems Engineering (IMTEK), University of Freiburg, Germany. He was the Bioanalytical Scientist at Bristol-Myers Squibb Company, Dublin, Ireland (2006-2009) and the Team Leader at NUS Nanoscience and Nanotechnology Institute, Singapore (2009-2012) after completing his Ph.D. from Central Scientific Instruments Organisation, India (2006). His research outputs include many successful technology transfers, several in vitro diagnostic and point-of-care technologies, over 80 publications in high impact factor journals and numerous patents, book chapters, and publications/talks in international conferences. He has constantly received highly prestigious fellowships and awards for scientific research excellence and innovation. Besides, he is acting as the Editorial Board and Executive Editor for several reputed journals (such as NPG’s Scientific Reports, Elsevier’s NPBR and MDPI’s Diagnostics). His research interests include in vitro diagnostics, biosensors, point-of-care testing, nanotechnology, and smartphone-based personalized healthcare monitoring and management.
Abstract:
Several proprietary Smartphone-based colorimetric readers (SBCRs) for in vitro diagnostics (IVD) have been developed using a standard Smart phone (Samsung Galaxy SIII mini), a gadget (iPAD mini, iPAD4, iPhone 5S or any similar tablet/Smartphone), a custom-made dark hood and a base holder assembly. A 96 or 24-well microtiter plate (MTP) was positioned on the g adget’s screensaver, enabling white light-based bottom illumination only at the well bottom. The set-up was demonstrated and compared with a commercial MTP reader (MTPR) for human C-reactive protein immunoassay and bicinchoninic acid (BCA) protein estimation assay. Colorimetric images of the reaction solutions were captured by the Smartphone’s back camera with 5 megapixels resolution. The pixel int ensity of the captured images was determined by an image processing algorithm. SBCR has same precision, dynamic range and sensitivity as MTPR. SBCRs would have a considerably commercial opportunity as immunoassays using the clinical IVD kits are based on color detection. Being 20-fold cheaper than MTPR, SBCR could be an ideal point-of-care (POC) colorimetric detection device for the next-generation of cost-effective IVD and versatile tools in bio ana lytical settings. Moreover, they could be employed for colorimetric assays in the visible range using a transparent and flat bio analytical platform such as lab-on-a-chip, lateral flow microfluidics and centrifugal microfluidics-based LabDisk platforms. Considering their widespread use (~7 billion users, ~96% world population), Smartphone’s are under on-going improvement to become smarter, smaller and cheaper. As more and more objects are linked to with a Smartphone, SBCR will become a powerful assistant of analysts to play an important role in IVD, POC, personalized healthcare monitoring and management, telemedicine and remote bio sensing applications.
Keynote Forum
Aline Zimmer
Merck Millipore Germany
Keynote: Chemically modified cysteine in fed-batch processes and impact on CHO specific productivity
Time : 10:00-10:30
Biography:
Based in Darmstadt, Germany, Aline is currently responsible for the development of mammalian cell culture media at Merck Millipore. Aline holds an engineering degree in biochemistry and biotechnology from the National Institute of Applied Sciences (INSA) in Lyon, France, a master in cellular physiopatho logy from the University of Lyon and a PhD degree in immunology from the University of Paris. She has worked 3 years in immunotherapy biomarker research at Stallergenes before joining Merck KGaA in 2011. From 2011 to 2012, Aline was responsible for the development of cellular assays to monitor upstream processes at Merck Millipore. From 2013 to 2014, Aline was heading the R&D upstream media performance laboratory responsible mainly for the development of new chemically modified amino acids and their integration in fed-batch platforms. Since April 2014, Aline is heading the mammalian cell culture media development activities at Merck Millipore.
Abstract:
Industrial fed-batch cultivation of mammalian cells is used for the production of therapeutic proteins such as monoclonal antibodies. Beside medium ensuring initial growth, feeding is necessary to improve growth, viability and antibody production. Established commercial systems include a slight acidic concentrated main feed and a separate alkaline amino acid feed containing L-tyrosine and L-cysteine. Since L-cysteine is not s table due to its dimerization to cystine in the presence of air and metal catalysts, a stable L-cysteine derivative is needed to include all amino acids in neutral pH feeds. Those single feed systems are favored to simplify feeding schemes and improve the overall process robustness through stabiliz ation of both pH and DO signals. Here, we suggest the use of a chemically modified cysteine in combination with phosphor tyrosine disodium salt in an industrial single feed fed-batch process applicable at small scale as well as in bioreactor s. Cell culture experiments were carried out either in spin tubes or bioreactors with a CHO suspension cell line expressing a human monoclonal antibody. Viable cell density and viability were measured using an automatic cell counting device. Spent media analysis of supernatants was carried out for amino acids after pre-column derivatization and UPLC analysis and for vitamins using LC-MS/MS. Metabolite measurements were performed with Cedex Bio HT relying on photometric and turbidometric methods. Characterizati on of the monoclonal antibody was performed using 2-AB labeling for glycan analyses, cIEF for charge variants analyses and LC-MS/MS for peptide mapping experiments. Stability studies of the feed containing the modified cysteine derivative showed that the molecule was stable and that no L-cysteine or L-cystine was released over three months when stored at room temperature or 4 ° C. Moreover, no change in the color of the feed was observed over time. Small scale batch experiments where L-cysteine was replaced by the same amount of chemically modified cysteine indicated no change in growth or viability profiles. Use of the modified cysteine derivative in small scale fed-batch processes indicated comparable maximum viable cell density, prolonged viability and increased titer compared to the established two feed system. Bioreactor experiments confirmed the increase in specific productivity described at small scale when the single feed strategy was compared to the two feed strategy. In depth characterization of the monoclonal antibody indicated no change in the glycosylation or charge variant pattern whereas peptide mapping experiments were not able to detect any integration of the modified amino acid in the sequence of the monoclonal antibody.
Keynote Forum
Sushanta K. Mitra
Professor & Chair of Mechanical Engineering, York University Canada
Keynote: Micro & Nanofabrication for Biosensors
Time : 10:30-11:00
Biography:
Sushanta Mitra is a Kaneff Professor in Micro & Nanotechnology for Social Innovation at the York University, Toronto. His research interests are in the fundamental understanding of fluid flow in narrow confinements and its applications in the fields of energy, environment and bio-systems. For his contribution in science and engineering, he has been an elected fellow of the American Society of Mechanical Engineers, the Canadian Society for Mechanical Engineering, the Engineering Institute of Canada, the Canadian Academy of Engineering and the Royal Society of Chemistry.
Abstract:
In recent years there is a significant development in micro and nanotechnology that has enabled fabrication of fine micro/nanostructures, which are often used as building-block towards producing sensors which are specific, selective and sensitive towards number of key biomolecules. The talk will focus on how such micro/nanofabrication tools coupled with fundamental developments in physics and chemistry can create next generation sensors that can be used for monitoring deadly virus like Dengue NS1, food borne pathogen like Listeria monocytogenes, water borne pathogens like E. coli and cardiac markers like myoglobin for early diagnostics of heart attack. The talk will end with example of a translational and transformative research which allows the laboratory proto-type to be tested in field and end-user communities.
- Industrial Aspects of Biotechnology
Location: Flemings Conference Hotel Room 7
Session Introduction
K Yankulov
University of Guelph, Canada
Title: Novel synthetic anti-microbial defensins through confrontational selection and screening of yeast libraries
Time : 11:45-12:10
Biography:
K Yankulov is a Professor at the University of Guelph, Canada. His lab is using the yeast S. cerevisiae for research in several closely related areas of extra-chromosomal DNA , protein expr ession and the design and screening of expression DNA library screening . This line of research has produced 33 publicatio ns on genes in influential genetics journals. He is an Editor in PLoS One, Frontiers in Genetics and Genes and Gene Therapy. Recentl y, he has pursued his interest on a group of novel fungal defense molecules (called defensins) and their commercial development as anti-microbial agents
genetics , including the stability ofAbstract:
food borne pathogens. They have a low probability of developing microbial resistance and are viewed as viable alternatives to antibiotics for both the food and pharmaceutical industries. The industrial development of vertebrate defensins is hindered by concerns of cytotoxicity. In contrast, the known fungal defensins show little side effects in animals and work at doses comparable to these of many antibiotics. We are developing a platform that will use DNA libraries for a huge variety of synthetic defensin-like peptides. We are expressing these libraries in yeast and screen them against model microorganisms. Our aim is to identify novel synthetic anti-microbial peptides that can be used as alternatives to antibiotics. These agents can be improved to incorporate a trypsin-target site to ensure their normal degradation in the stomach and to abolish any side effects on the normal microbiome of the animals. We also aim at the affordable production of such agents by the yeast K. lactis in milk whey. Our progress in these screens will be reported and discussed.
Antibiotics are frequently used in animal feed to boost its efficacy. However, recent policies in EU and USA have restricted s uch use of antibiotics and Canada is expected to follow suit. Consequently, the need for novel anti-microbial agents is a most urgent issue. Defensins form a large group of secreted animal, plant or fungal peptides that kill a broad spectrum of microorganisms includin gFuad Fares
University of Haifa, Israel
Title: Long acting recombinant glycoprotein hormones: From bench to clinics
Time : 12:10-12:35
Biography:
Fuad Fares has completed his DSc studies at the Faculty of Medicine, Technion-Israel Institute of Technology and Postdoctoral studies at the Department of Molecular Biology and Pharmacology, School of Medicine, Washington University, St. Louis Missouri. He is the Director of the Department of Molecular Genetics at Carmel Medical Center and Associated Professor at the Department of Human Biology, University of Haifa. He has published more than 75 papers in reputed journals and serving as a member of the Israel Council for Higher Education. He is the inventor of designing long-acting recombinant proteins and the initiator of PROLOR Biotech Company.
Abstract:
One major issue regarding the clinical use of many peptides is their short half-life due to the rapid clearance from the circulation. To overcome this problem, we succeeded to ligate the signal sequence of O-linked oligosaccharides to the coding sequence of the hormones. The cassette gene that has been used contains the sequence of the carboxyl-terminal peptide (CTP) of human chorionic gonadotropin ï¢ (hCGï¢) subunit. The CTP contains 28 amino acids with four O-linked oligosaccharide recognition sites. It was postulated that O-linked oligosaccharides add flexibility, hydrophilicity and stability to the protein. On the other hand it was suggested that the four O-linked oligosaccharides play an important role in preventing plasma clearance and thus increasing the half-life of the protein in circulation. Using this strategy we succeeded to ligate the CTP to the coding sequence of follitropin (FSH), thyrotropin (TSH), erythropoietin (EPO) growth hormone (GH) and thus to increase the longevity and bioactivity of these proteins in vivo. Interestingly, the new analogs of FSH and GH were found not immunogenic in human and it is already passed successfully clinical trials phase III and phase II respectively. Moreover, FSH long acting was approved by the European Commission (EC) for treatment of fertility. In addition, our results indicated that long acting GH is not toxic in monkeys and the results from clinical trials phase I and phase II seem to be promising. Designing long acting peptides will diminish the cost of these drugs and perhaps reduce the number of injections in the clinical protocols.
Tarek M Mohamed
Tanta University, Egypt
Title: Immobilization and characterization of inulinase from Ulocladium atrum on nonwoven fabrics
Time : 12:35-13:00
Biography:
Tarek had born in Egypt, Cairo on 1966. Tarek is a Professor of biochemistry in Biochemistry Division, Chemistry Department, Tanta University, Tanta, Egypt started from June 2008 till now. He is associated professor In Tanta University fromm 2003-2008. He worked in National Research Centre from 1992 to 2003 in enzyme engineering , biochemistry.
Molecular Biology Department as a researcher. Tarek is a reviewer for many journals. He shared in 12 national projects. He Supervised more than 20 MSc a Ph.D. thesis. He has 42 publications. Tarek interested in Purification and characterization of enzymes , enzyme inhibitors . Study relatedAbstract:
Ulocladium atrum inulinase was immobilized on different composite membranes composed of chitosan/nonwoven fabrics. Km values of free and immobilized U. atrum inulinase on different composite membranes were calculated. Free and immobilized U. atrum inulinase had pH optima at 5.6 on polyester nonwoven fabric coated with 3% chitosan solution (PPNWF3) but immobilized U. atrum inulinase on polyester and polypropylene nonwoven fabrics coated with 1% chitosan solution had pH optimum 5. The enzyme had optimum temperature at 40° C for immobilized enzyme on each of polyester and polypropylene composite membranes coated with 1% chitosan meanwhile it was 50° C for free and immobilized enzyme on polypropylene nonwoven fabric coated with 3% chitosan solution. Inulinase was stable at 40° C for free U. atrum inulinase but thermal stability of immobilized enzyme was detected up to 60° C. Reusability of immobilized enzyme was from 38 to 42 cycles of reuse; the immobilized enzyme lost its activity completely. In conclusion, immobilized U. atrum inulinase was considerably more stable than the free enzyme and could be stored for extended periods.
Long Liu
Jiangnan University, China
Title: Systems metabolic engineering of Bacillus subtilis for efficient N-acetyl glucosamine production
Time : 13:45-14:10
Biography:
Long Liu is currently a life-time professor at School of
Biotechnology , Jiangnan University, Wuxi, China. He has been working in the area of bioprocess engineering and metabolic engineering with special reference to bioprocess optimi zation and control. He has authored 2 books, 3 book chapters, 7 review papers, 20 patents, 37 research papers in SCI journals, and 10 conference papers. He has been a recipient of First pri ze of Science and Technology progress, Jiangsu, China (2010), First prize of Science and Technology Progress, China Petroleum and Chemical Industry Federation (2011), the Technological Invention Award of China National Light Industry Council (2013), and Excellent Young Teacher of Jiangsu, China (2014).Abstract:
N-acetyl glucosamine (GlcNAc) is a pharmaceutically and nutraceutically important compound with wide applications and now is mainly produced by hydrolysis from crab and shrimp shells which can cause severe environmental pollution and has potential risk of allergic reactions. In this work, we achieved the over-production of GlcNAc by systems metabolic engineering of Bacillus subtilis, a generally regarded as safe strain. Specifically, GlcNAc synthesis pathway was strengthened by co-over expression of Glucosamine-6-phosphate (GlmS) synthase and GlcNAc-6-phosphate N-acetyl transferase (GNA1) which realized GlcNAc production (240 mg/L). Next, GlcNAc uptake pathway and intracellular degradation pathway were entirely blocked by knockout of all the encoding genes in GlcNAc catabolic pathway to facilitate GlcNAc accumulation. Then, to balance and strengthen GlcNAc synthetic pathway, DNA-guided scaffold system was introduced and increased GlcNAc titer from 1.83 g/L to 4.55 g/L. Synthetic small regulatory RNAs were then employed to optimize expression level of key enzymes in the nodes of GlcNAc-related network including 6-phosphofructokinase (Pfk) and phospho glucosamine mutase (GlmM). GlcNAc titer was improved to 8.30 g/L by modular regulation of the activities of GlcNAc-related modules. In fed-batch fermentation, the GlcNAc titer was further increased to 31.65 g/L which was 3.8-fold that in the shake flask. Finally, to understand kinetics of metabolite changes in GlcNAc synthesis pathway and glycolysis, targeted metabolomics and dynamic labeling were implemented. Inefficient GlcNAc6P dephosphorylation and undesired GlcNAc phosphorylation were identified as rate-limiting step for GlcNAc synthesis which pinpointed future direction for further pathway optimization. The used systems metabolic engineering strategies may be useful for the construction of versatile B. subtilis cell factories for the production of the other industrially important chemicals.
Jingwen Zhou
Jiangnan University China
Title: Enhancing flavonoid production by systematically tuning the central metabolic pathways based on a CRISPR interference system in Escherichia coli
Time : 14:10-14:35
Biography:
Prof. Jingwen Zhou obtained his Ph.D degree in Fermentation engineering in 2009. After that, he became assistant professor (20 09), associate professor (2011) and full professor (2014) in School of Biotechnology, Jiangnan University. He finished his postdoc training in Department o f Chemistry and Chemical Biology in Harvard from 2012 to 2013. His current research works mainly focused on the metabolic engineering of microorganisms to produce organic acids and plant natural products, especially L-ascorbic acid and flavonoids. He published 52 peer reviewed papers on journals such as Metaboli, Applied and Environment Microbiology, and also several invited reviews on Current Opinion in Biotechnology and Biotechnology Advances. Several of the typical products he had been working on were now produced by several manufactures on industrial scale. His achievements were awarded for several times inside China. He is now the Editorial Board of Scientific Reports (Nature Press) and Electronic Journal of Biotechnology (Elsevier Press). c Engineering
Abstract:
The limited supply of intracellular malonyl-CoA in Escherichia coli impedes the biological synthesis of polyketides , flavonoids and biofuels. Here, a clustered regularly interspaced short palindromic repeats (CRISPR) interference system was constructed fo r fine-tuning the central metabolic pathways to efficiently channel carbon flux toward malonyl-CoA. Using synthetic sgRNA to silence candidate genes, genes that could increase the intracellular malonyl-CoA level by over 223% were used as target genes. The efficiencies of repression of these target genes were tuned to achieve appropriate levels so that the intracellular malonyl-CoA level was enhanced without significantly altering final biomass accumulation (the final OD600 decreased by less than 10%). Based on the results, multiple gene silencing was successful in approaching the limit of the amount of malonyl-CoA needed to produce the plant-specific secondary metabolite (2S)-naringenin. By coupling the genetic modifications to cell growth, the combined effects of these genetic perturbations increased the final (2S)-naringenin titer to 421.6 mg/L which was 7.4-fold higher than the control strain (50.5 mg/L). The strategy described here could be used to characterize genes that are essential for cell growth and to develop E. coli as a well-organized cell factory for the production of other important products that require malonyl-CoA as a precursor such as flavonoids, polyketides and fatty acids.
Guocheng Du
Jiangnan University, China
Title: One-step biosynthesis of α-keto acids by the L-amino acid deaminase: Biocatalyst construction and process optimization
Time : 14:35-15:00
Biography:
Guocheng Du is the Dean of School of bioprocess engineering and metabolic engineering. He has about 300 publi cations/communications, which include 68 patents, 10 books, 176 original and review papers, etc. He won two Second Prizes of State Science and Technology Awards in 2006 and 2012, respectively. He has been conferred Honorary Doctorate degree from Jiangnan University, China. He is an Associate Editor of Journal of the Science of Food and Agriculture, and Microbial Cell Factories, and Board Member of Bioresource Technology
Biotechnology , Jiangnan University, China. He is the Distinguished Professor of Changjiang Scholars, Ministry of Educat ion, China. His current main research focus is onAbstract:
L-Amino Acid Oxidases (LAAOs ) which catalyze the stereo-specific oxidative deam ination of L-amino acids to α-ketoacids and ammonia are flavinadeni ne dinucleotide containing homodimeric proteins. L-Amino acid oxidases are widely distributed in diverse organisms and have a range of properties. In this work, we systematically engineered the LAAO from Proteus vulgaris as a robust biocatalyst for the efficient one-step biosynthesis of a-keto acids including α-ketoglutaric acid, α-keto-γ-methyl thiobutyric acid and phenyl pyruvic acid which are currently produced by multi-step chemical synthesis with heavy environmental pollution. First, the directed evolution of LAAO was done by error-prone Polymerase Chain Reaction (PCR) and then the site-saturation mutagensis was conducted at the mutation sites of the positive mutants yielded by error-prone PCR. The biotransformaton conditions included biocatalyst concentration, the substrates concentration, temperature and pH and so on. The maximum titers of α-ketoglutaric acid, α-keto-γ-methyl thiobutyric acid and phenyl pyruvic acid are 12.3 g/L, 63.6 g/L and 11.8 g/L, respectively under the specific optimal conditions. Compared with the traditional multi-step chemical synthesis, our one-step biocatalytic production of a-keto acids has an advantage in terms of less environmental pollution and thus has great potential for industrial production of a-keto acids.
Reichelt W N
Vienna University of Technology, Austria
Title: Transient physiologic states: Friend or foe?
Time : 15:00-15:25
Biography:
Reichelt W N is a Project Assistant in the Christian Doppler Laboratory for Mechanistic and Physiological Methods for Improved Bioprocesses at the Vienna University of Technology (VUT). He studied Biochemistry at the Karl Franzens University in Graz before doing his Master’s thesis in the field of programmed cell death and aging. After a year of PhD research in the field of neurodegeneration (Parkinson), he transitioned to the field of bioprocess technology at VUT to acquire in-depth understanding of pharmaceutical bioprocess production process approaches. Within his PhD work, he is spanning applied to basic science by developing and employing physiological bioprocess control strategies to tweak industrial bioprocesses to maximum productivity. Hereby, the project unifying goal is to gain mechanistic insight into bioprocess physiology to ultimately achieve comprehensive bioprocess understanding and transferable platform knowledge.
Abstract:
Progressively expiring patent protection brings forward generic drug industry and heats up competition for the most efficient and robust bioprocesses. While competition is intensifying, the FDA quality by design (QbD) initiative for increased process understanding is increasing demands concerning bioprocess development. These circumstances cumulate to the necessity of comprehensive process understanding and consequently highly efficient bioprocess development routines. For bioprocess development, literature has comprehensively proven the feeding strategy an especially promising target. Having mastered the feedback control of physiological rates by the use of soft sensors, the subsequent step of biotechnological development could lead towards controlled transient changes of physiological parameters. Whereas the majority of the scientific community has chosen the specific growth rate (µ) as target control parameter, we have invested extensive work in the control of the specific substrate uptake rate (qs). Hereby, the qs control renders physiology accessible, substrate availability being upstream of physiological variables example µ. We want to test the hypothesis that dynamic process control as qs profiles in terms of ramps and oscillations constitute tools for efficient tweaking of physiology. By alternating the substrate availability in the reactor the physiologic state of E.coli shall be modulated between high metabolic load and recreational phases during induction. Focusing on product physiology interrelations, modeling and investigation of scale up effects is explicitly not in scope of our investigation. Hereby, we want to address specific questions as the impact frequency and amplitude for qs oscillation of on physiology as well slope and orientation of qs ramps. The amplitude of oscillation is of great interest can physiological limits be temporarily exceeded by granting periods of recovery? Whereas a high frequency of oscillation of the specific substrate uptake rate could be limited by technical constraints (example sampling capacities, error of measurement) a too long duration could fail to have any impact besides acetate formation.
C J Andrade
State University of Campinas, Brazil
Title: Surface activity of surfactin under extreme conditions (pH, temperature and ionic strength): A prospection for microbial enhanced oil recovery
Time : 15:25-15:50
Biography:
C J Andrade is a PhD student on Biotechnological Processes at State University of Ca mpinas, and has been trying to use industrial wastes as a culture medium . Among the compounds of his interest are biosurfactants, galacto-oligosaccharides, enzyme, flavours, etc. He is also interested on the integration between fermentation and purification, in particular, ultrafiltration.
Abstract:
Petroleum has been playing the most important role in energy sectors. Worldwide oil production has been declining. The primary and secondary techniques of oil recovery, reach ≈35% and ≈20%, respectively. Thus, the aim of enhanced oil recovery technologies is the residual oil, which is ≈45% of the total oil reserves. Microbial-enhanced oil recovery (MEOR) is one of the most important tertiary recovery processes, which rise up the oil recovery ≈3%. However laboratory scale experiments predict up to 10%. The aim of this work was to evaluate the surface activity of surfactin under extreme conditions (pH, temperature and ionic strength) - individually and their interactions – which represents similar environmental of wells. The surface activity of surfactin under extreme condition was already reported. Although, for the best of our knowledge this is the first study that focused on the biosurfactants surface activity under extreme condition at the same time, which is fundamental for MEOR. Surfactin was produced at bioreactor scale (7.5L) using cassava wastewater as substrate. Then, surfactin was collected from the top of bioreactor centrifuged, precipitated (pH 2) and purified by solvent extraction and ultrafiltration. The final product was named produced surfactin and used in the experiments at 74 mg.L-1 of surfactin. The ionic strength was tested using synthesized brine. The pH, temperature and ionic strength were first investigated separately, in which the pH was evaluated from 2 to 12, whereas 3 temperatures were tested during 1h, 79, 100 and 121oC and finally the ionic strength 2.5, 5, 10 and 20 g.L-1 of synthesized brine. Then, were measure surface tension (ST) and its critical micelle dilutions (CMD). Thereafter, central composite design (CCD) experiments evaluated the interactions of temperature, pH and ionic strength. The ionic strength affected the surface activity property of surfactin, in which the higher ionic strength, the higher was the surface tension measurements (ST and CMD). The intermediate pH (4-8) showed highest (best) surface activity. No significant differences were observed for the thermal treatments. Therefore, temperature, pH and ionic strength experiments were aligned with the maintenance of surface activity properties of produced surfactin, which suggest their potential application at extreme systems, in particular MEOR. Regarding to CCD of surfactin, the ANOVA of CMD-1 indicated that temperature, pH and ionic strength were statistically different (95%) with r2 of 0.76. Thus was generated the equation 1: [Y = 67.28 + 8.5 x (ionic strength) – 3.02 x (ionic strength)2]. The ionic strength was the most significant parameters on the surface activity, in which threshold of surface activity of surfactin was at 18.58 g.L-1. The validation (72.17 mN.m-1) was well-aligned to the predicted value (73.26 mN.m-1). Therefore, it seems feasible consider surfactin in MEOR, in which ionic strength is the most important parameter.
- Agriculture Biotechnology
Location: Flemings Conference Hotel Room 7
Session Introduction
Raoufa A I Abdel Rahman
Genetic Engineering and Biotechnology Research Institute, Egypt
Title: Production of rosmarinic acid from in-vitro culture of Basil (Ocimum basilicum L) and lemon balm (Melissa officinalis L)
Time : 09:30-09:55
Biography:
Raoufa A I Abdel Rahman has completed her PhD in 2003 at Alexandria University, Faculty of Agriculture, Pharmaceutical Bio-products Research Dep ., City of Scientific Research and Technology Applications, Alex., Egypt. Her main research interests are using plant tissue culture and genetic engineering techniques to conserve rare and endangered plant species, as well as enhancement in the productivity of important pharmaceutical compounds from plants. She is working in several projects dealing with the production of antiviral , anticancer and antioxidant compounds f rom plants using in-vitro cultures.
Genetics Dep., through a collaborative scholarship with University of Georgia, Athens, USA. She completed a research project entitled "Exploring IRES mediated discistrons for the phytoremediation of Mercury". Currently, she is the Director ofAbstract:
Over the past three decades, plant cell and tissue culture and plant biotechnology techniques have proved to be a valuable tool to study bio synthesis and production of plant secondary products. Rosmarinic acid (RA) is a natural phenolic compound produced by variety speci es of Lamiaceae and Boraginaceae families. In the recent few years, RA has attracted attention due to its antiviral, antibacterial , anti-inflammatory and anti-aller gic activities. In the present study, the efficiency of us ing plant tissue culture techniques as a promising alternative method for the production of RA from basil and lemon balm was tested. Callus and shoot cultures were established using different types of explants and medium composition to test the optimum conditio n for their production. Results showed that leaf explants were the best explants for callus production. MS medium containing 1mg/l BA and 1mg/NAA (MS2) was the best medium for callus growth in lemon balm while MS4 was the optimum media for basil. Stem segments and shoot tips were used as explants for production of shoot cultures in lemon balm and basil. RA levels were determined in control, callus and shoot cultures. Results showed that callus and shoot cultures accumulated higher levels of RA compared to control. RA levels ranged from 9.42±1.27 to 38.25±0.73 ug/ mg plant dry weight. Shoots produced 3 and 2.7 folds higher rosmarinic acid levels compared to control in lemon balm and basil. The overall results indicated that plant tissue culture is an efficient tool for RA production from basil and lemon balm plants.
S. K. Tiwari
State Forest Research Institute, Jabalpur India
Title: Standardization of clonal macropropagation protocol of Dillenia pentagyna roxb an important and endangered medicinal tree species through stem branch cuttings
Time : 09:55-10:20
Biography:
Dr. Shailendra Kumar Tiwari has completed his PhD at the age of 29 years from Rani Durgawati University, Jabalpur. He is the Senior Scientist of State Forest Research Institute, Jabalpur a premier Forestry Research service organization. He has published more than 40 papers in reputed journals. He was the member of Society of In vitro Biology, USA, since 2013. He has more than 28 years of research experience in Forest Genetics, Forest Tree Improvement, Plant Propagation particularly Macro and Micropropagation and Plant Biotechnology. He trained more than 2000 students and other stake holders in the field of plant tissue culture and plant biotechnology. He has also Visited New Zealand as FAO Fellow in 1998 and presented paper in 2007 in SIVB conference at Indianapolis, USA.
Abstract:
Abstract: Dillenia pentagyna belongs to family Dilleniaceae , is commonly known as Karkat. It is small deciduous tree reaching up to the height of 10-12mt with a straight bole. The species is distributed throughout in India including Andaman & Nicobar. It is highly important medicinal tree species. Its leaves, fruits and bark shows antibacterial , anti-alpha glucosidase and antioxidant property. Due to its hi gh medici nal value the natural occurrence of this species is declining day by day and gradually the species comes under the threat condition. The species is highly recalcitrant in nature, because it very difficult to propagate by conventional propagation methods. Now there is an urgent need to develop an appropriate technology for its conservation and clonal propagation. An attempt has been made for standardizing the clonal propagation technique of this valuable medicinal species through stem branch cuttings under intermittent misting conditions in mist chamber. The optimum rooting response has been standardized by various PPM concentrations and treatment timings of root promoting hormones IBA and NAA. The optimum rooting response > 60 percent was observed when the cutting were treated up to 20 minute with 500 ppm concentration of IBA. On an average 6 roots with10-15cm length were induced from the cuttings after 30-35 days.NAA was failed to induce roots from the cuttings
Muhammad Asgher
University of Agriculture, Pakistan
Title: Activity and thermostabilty improvement of Trametes versicolor IBL-04 laccase by chitosan beads immobilization
Time : 10:20-10:45
Biography:
Abstract:
The laccase bioremediation and biosensor development. Monomeric 66 kDa laccase produced by Trametes versicolor IBL-04 in SSF of corncobs (911 U/ mL) was purified through ammonium sulphate precipitation, dialysis, ion-exchange chromatography and gel filtration. The purified laccase was immobilization using chitosan as support material and glutaraldehyde as activator/cross linking agent. Chitosan concentration of 2.5% was optimum for preparation of most stable 2.0 mm size chitosan beads activated by 1.5% glutaraldehyde for best laccse immobilization. Scanning electron microscopy showed that beads with immobilized laccase on the surface were spherical in shape having large surface area. The immobilized laccase was found to be catalytically more vigorous and stable and it worked over a wide range of pH 3-6 and temperature 45-65o C. Laccase immobilized with chitosan beads had 936 U/mL at pH 6 and 60° C and improved thermal behavior. The chitosan beads immobilized laccase had higher Km (93 µM) and V-max 944 µM/min values as compared to its soluble counterpart thus demonstrating its higher catalytic efficiency. The kinetic and thermostability characteristics of chitosan beads immobilized laccase reflect that the enzyme has potential for use in industrial and environmental biotechnology.
enzyme has got tremendous applications in diverse industrial pr ocesses including biomass delignification, bio pulping in paper and pulp, as denim stone wash agent, detergents,Anjali Jaiwal
University of Delhi, India
Title: Silencing of hormonal biosynthesis genes by double stranded RNA (dsRNA) impairs larval growth and development of cotton bollworm (Helicoverpa armigera)
Time : 10:45-11:10
Biography:
Myself Anjali Jaiwal, I am pursuing Ph.D under the supervision of Prof. M.V. Rajam, Head Dept. of gene sequences to GenBank of NCBI. Published one review article ‘Coenzyme Q10 production in plants: current status and future prospects’ in ‘Critical reviews in Biotechnology’ as second author. Attended six national and three international conferences. Awarded CSIR-UGC JRF and prestigious DST-INSPIRE fellowship by the DST (Department of Science and Technology). My research work includes the screening of few vital genes of Helicoverpa armigera by feeding target gene dsRNAs to the insect pest via semi-synthetic artifici al diet. Also, development of insect resistant transgenic tobacco and cotton plants via Plant-mediated RNAi silencing of vital genes of H. armigera
Genetics , University of Delhi South Campus, New Delhi, India. I did my post-graduation in Biotechnology from M.D. university, Rohtak, Haryana and received Gold medal for standing first in the university. Also ranked first among the science students during graduation. Received different scholarships during graduation and Post-graduation. Cloned and submitted threeAbstract:
Helicoverpa armigera is a polyphagous insect pest responsible for major losses in cotton and other agronomically important crops. RNA interference (RNAi) has emerged as a potential alternative to raise insect-resistant plants by in planta expression of dsRNA specific to a vital insect gene. In the present study, the hormonal biosynthesis genes in H. armigera were targeted by feeding dsRNAs correspond in g to each target gene viz., juvenile hormone acid methyl transfererase (HaJHAMT), prothoracicotropic hormone (HaPTTH), pheromone biosynthesis-activating peptide (HaPBAP), molt regulating transcription factor (HaHR3), activated protein 4 (HaAP-4) and eclosion hormone precursor (HaEHP) which play key roles in regulation of physiological, developmental and behavioural events in the target insect pest. Ingestion of target gene dsRNAs via artificial diet resulted in variable mortality ranging from 60-92% in all the six targeted genes. Silencing of the target genes showed retarded larval growth, delayed in molting, metamorphosis and pupal formation. A comparison of the silencing potency of un-diced long HaPTTH dsRNA with RNase III-diced-siRNAs revealed that long dsRNAs were more effective in target gene silencing as compared to siRNAs. The HaPTTH-dsRNA coated onto the detached leaf was found to be more effective in silencing target gene when compared to dsRNA feeding via artificial diet. The qRT-PCR analyses showed that mRNA level of six target genes was drastically reduced compared to control or unrelated GFP-dsRNA control cor related with the developmental defects. These results indicate that hormonal biosynthesis genes can be used as vital targets for improving pest resistance in cotto n and other crop plants which are infested with H. armigera.
Khaled Hussein Arafat
Assiut University Egypt
Title: Biological control of Erwinia amylovora on pear trees by alternative bactericides
Biography:
Abstract:
The blossom blight phase of fire blight disease on pear trees, caused by the bacterium Erwinia amylovora (Burrill), is typically managed by applying the antibiotic , streptomycin sulfate and copper, to trees during blossom. Biological control agents of fire blight can be achieved by applying nonpathogenic bacteria, viz. Bacillus subtilis or Pantoea agglomerans and plant extract, viz. Harmel during open flowers as spraying treatments. The objective of this study was to examine the alternative bactericides against bacterium E. amylovora in vitro and in vivo during two seasons (2013-2014) in Al-Gharbia governorate, Egypt. Our results confirmed the ability of these non-pathogenic bacteria and plant extract can decreased fire blight severity on pear trees. Further studies with different locations in Egypt and larger sample sizes would allow us to make stronger recommendations including their ability to prevent disease and make it in integrated pest management program.
- Track 1: Biotechnology in Healthcare
Location: Flemings Conference Hotel Room 7
Session Introduction
Guido Krupp
Amptec GmbH, Germany
Title: Synthetic mRNAs present a rapidly growing technology: Optimised tool for stem cell generation and for manipulating cellular phenotypes
Time : 12:00-12:25
Biography:
Guido Krupp is the CEO and President of Amptec GmbH. He received his PhD degree from Würzburg University & Max-Planck-Institute, Martinsried. He did his Post-Doctoral at Yale University. He is designated as a Research Group Leader at Kiel University. He is the Founder of Artus GmbH & Amptec GmbH. His research interests include nucleic acid technology with focus on RNA, plant pathogens (viroids), ribozymes and telomerase. He has published more than 60 publications and has been designated as an Editor of Ribozyme Biochemistry & Biotechnology, and of Telomeres, Telomerases & Cancer, and Editorial Board Member of Biotechnology Annual Review.
Abstract:
Availability of high quality synthetic mRNAs (syn-mRNAs) has enabled progress in their applications. Growing interest of private investors and big pharma has created a novel billion $ business. A rare situation was there in which two German enterprises are among the three top players in the field. Amptec recognizes its obligation to support new players by providing customized, high quality mRNA products. Requirements in the application of mRNA-mediated manipulation of cells were (i) expression of antigens in dendritic cells for vaccination in oncogenesis, infectious disease and allergy prevention; (ii) reprogramming of fibroblasts to induced pluripotent stem cells with subsequent differentiation to the desired cell type; (iii) applications in gene therapy. A recent overview presents applications and corresponding syn-mRNA quality requirements. Syn-mRNAs can be generated by In-Vitro Transcription (IVT) from templates containing the synthetic gene. In principle, linearized plasmids can be used as templates. However, this procedure is hampered by several disadvantages such as incomplete plasmid cleavage resulting in poor reproducibility; and high amounts of plasmid DNA introducing undesired bacterial components with possible complications of in-vivo applications. Furthermore, optimal mRNA activity depends on a very long, unmasked poly (A) tail, ideally 120 nucleotides long. But, long homo-polymeric repeats are unstable in bacterial cells. We have developed an alternative procedure, with well defined PCR products as IVT-templates. This approach and detailed quality requirements for synthetic mRNAs are presented. Problems which were observed in IVT-based mRNA synthesis are shown, combined with problem solutions.
Soha M Hamdy
Fayoum University, Egypt
Title: Biochemical Studies on the Effect of Turmeric on Breast Cancer of Rats
Time : 12:25-12:50
Biography:
Soha Mohamed Hamdy has completed her PhD in 2001 from Fayoum University. Presently, she is a Professor in Faculty of Science - Chemistry Department at Fayoum University, Egypt
Abstract:
Chemoprevention is regarded as one of the most promising and realistic approaches in the prevention of toxic effects of carcinogenic compounds. In this study, we investigated the chemoprevention efficacy of turmeric for 120 days against a single dose of (10 mg/rat) 7, 12-dimethylbenz (a) anthracene (DMBA). 60 rats were divided into four groups, 15 for each: Group I: Control; Group II: Injected with DMBA that induces mammary carcinoma; Group III treated with 5% turmeric before and after injection with DMBA; Group IV treated with 5% turmeric only as control 2 and the treatments were daily administered for 4 months. At the end of experiment the animals were sacrificed under anesthesia and their sera were used for evaluation; markers of tumorigenicity (serum levels of total sialic acid (TSA) and carcinoembryonic antigen); markers of endocrine derangement (serum prolactin and estradiol) and markers of oxidative stress (MDA for lipid peroxidation, nitric oxide and total antioxidant). The breast tissues were investigated for malignancy. Results showed statistical significant elevation of malondialdhyde (MDA), carcinoembryonic antigen (CEA), total sialic acid, prolactin, estradiol and nitric oxide also statistical significant decrease in body weight and total antioxidant in serum of DMBA treated rats as compared with control group but administration of turmeric was associated with decreased levels of tumorigenicity, endocrine derangement and oxidative stress. Histopathological examination revealed the formation of tumor in DMBA-induced rats and these abnormal changes were ameliorated in the rats supplemented with turmeric. In conclusion, these results suggested that supplementation of diet with turmeric provided antioxidant defense with chemopreventive activity against DMBA-induced mammary tumors.
Hattem M Mekky
University of Alexandria Egypt
Title: Biological activity of elicited Echinacea purpurea suspension cultures
Time : 13:50-14:15
Biography:
Lecturer, Department of Pharmacognosy, Faculty of Pharmacy, University of Alexandria and the organiser of the Biotechnology unit in the School of Pharmacy, University of Alexandria. Visitor lecturer for 3 months on yearly basis in Oman Assistant Pharmacists institute. Awarded the degree of Bachelor (1999) and Master (2003) in Pharmaceutical Sciences from The Department of Pharmacognosy, School of Pharmacy, University of Alexandria, Alexandria, Egypt in 2003. Awarded the degree of PhD (2009) in Tissue Culture and Genetic Engineering from The University of Nottingham, Sutton Bonington Campus, Leicestershire, LE12 5RD, UK. Research interests in phytochemistry, biosynthesis, pharmacology and biotransformation of chemicals using medicinal plants and the comparison between secondary metabolite profiles of wild plants and those produced by plant tissue culture.
Abstract:
Chitosan, phenyl alanine (Phe) and methyl jasmonate (MeJA) significantly increased the production of polyphenolics in suspension cultures of E. purpurea initiated on MS supplemented with 1.5 mg/L BA and 0.5 mg/L NAA. However, cultures elicited with 4 mM copper sulphate (CuSO4) possessed the highest EC100 in cytotoxicity experiments. Furthermore, Phe, MeJA, CuSO4 and chitosan significantly increased ABTS scavenging properties. Additionally, MeJA and Phe extracts stimulates the phagocytic and yeast intracellular killing activities of PMN and Finally, the anti-inflammatory properties of MeJA elicited cultures were the highest and were displayed by its strongest nitric oxide scavenging activity with an EC50 6.3 µg/ml and the most powerful inhibition of lymph proliferative activity induced by lipopolysaccharides with an EC50 4.5 µg/ml.
Yung-Chih Kuo
National Chung Cheng University Taiwan
Title: Rescuing apoptotic neurons from Alzheimer’s disease using wheat germ agglutinin-conjugated and cardiolipin-conjugated liposomes with encapsulated nerve growth factor and curcumin
Time : 14:15-14:40
Biography:
Dr. Yung-Chih Kuo is a professor at the Department of Department of Chemical Engineering, National Chung Cheng University. His research interests are focused on biomaterials, drug delivery system, tissue engineering, blood–brain barrier, stem cell differentiation, nerve regeneration, cancer therapy, Alzheimer’s disease treatment, biophysics, and colloid and interface science. In these fields, he has authored or coauthored over 100 SCI journal papers. He won Young Scholar Award in 2003 and Outstanding Research Award in 2010-13. He is also an associate editor of J. Taiwan Inst. Chem. Eng. (Impact factor 2.637) and an editorial board member in 11 international journals.
Abstract:
Liposomes with cardiolipin (CL) and wheat germ agglutinin (WGA) were developed to permeate the blood–brain barrier (BBB) and treat Alzheimer’s disease. WGA-conjugated and CL-incorporated liposomes (WGA-CL-liposomes) were employed to transport nerve growth factor (NGF) and curcumin (CUR) across a monolayer of human brain-microvascular endothelial cells regulated by human astrocytes and to protect SK-N-MC cells against apoptosis induced with β-amyloid1-42 (Aβ1-42) fibrils. An increase in the CL mole percentage in lipids increased the liposomal diameter, absolute value of zeta potential, entrapment efficiency of NGF and CUR, release of NGF, biocompatibility, and viability of SK-N-MC cells with Aβ1-42, however, decreased the atomic ratio of nitrogen to phosphorus and release of CUR. In addition, an increase in the WGA concentration for grafting enhanced the liposomal diameter, atomic ratio of nitrogen to phosphorus, and permeability for NGF and CUR across the BBB, however, reduced the absolute value of zeta potential and biocompatibility. WGA-CL-liposomes carrying NGF and CUR can be promising colloidal delivery carriers to target the BBB and inhibit neurotoxicity for future clinical application.
Essam F A Al-Jumaily
Baghdad University, Iraq
Title: Extraction and purification of extracellular glucosyltransferase (GTFs) from Streptococcus mutans serotype C isolate (H5)
Time : 14:40-15:05
Biography:
Prof. Dr. Essam Fadel A. Al-Juamily .Academic Qualification: Ph.D. degree: 1989, Biochemistry Dept. (Enzymology), Southampton University, U.K. Research Interests are Microbial Biotechnology; Enzyme Biotechnology; Biosafety and Biotechnology and Purification of Biotechnology materials with downstream production and Published 225 scientific research papers.
Abstract:
These isolates were tested for production of extracellular Glucosyltransferase (GTF) through determination of their enzyme specific activity. All isolates were able to produce the enzyme; Streptococci isolate (H5) which identified as Streptococcus mutans serotype C was selected as the best producible isolate for GTF with a specific activity of 2.6 U/mg. It was found that GTF of the chosen isolate (H5) was produced during the middle stationary phase (18-35 hours) and its maximal productivity was reached at 22 hours. Purification of S. mutans serotype (C) H5 GTF were done by ammonium sulfate, ion-exchange chromatography (DEAE-Sephacel column) and gel-filtration chromatography using Sepharose 6B column. The best percent saturation use for precipitating GTF by ammonium sulfate was 20-40% with specific activity 2.4 U/ml. Two purified GTF enzymes (GTF-I and GTF-II) were detected with specific activity 35.5 U/mg, 8.3 U/mg after 96.1 and 22.6 fold of purification respectively with yield 17.2%. Determination of purified GTF (GTF-I, GTF-II) molecular weight was done by using gel-filtration chromatography (sepharose 6B) column with presence of standards proteins. It was found that the molecular weight of GTF-I, GTF-II was 125819, 112201 Dalton respectively.
K Banu Köse
Istanbul Medipol University, Turkey
Title: The role of the patient specific vascular geometry and blood flow dynamics for vascular treatments
Time : 17:30-17:55
Biography:
K Banu Köse has published her MSc thesis on blood flow dynamics. She has published papers on medical imaging and computational blood flow dynamics. She is a PhD candidate of Biomedical Engineering at Istanbul Medipol University and Project Manager of Sidre Consulting about Cardiovascular Surgical Planning Solutions. She is teaching Medical Imaging Techniques at Istanbul Medipol University and is the Owner of the first website of Cardiovascular Mechanics Engineering News
Abstract:
Computational methods and three-dimensional imaging techniques have enabled the quantification of cardiovascular mechanics in vascular treatments especially for individual cases as congenital subjects. Biomechanical models based on multi-slice based on medical imaging, could provide more data about physiologic results of blood flow. The objective of this study is examining the mechanical effects of the blood flow which is relevant to the geometric topology of the arteries. The analysis includes tomography images of patients, codes of computer aided design and computational fluid dynamics. The realistic volume of the arteries of the patients obtained from DICOM by image segmentation methods. The geometry meshed by finite element models. The blood flow is recreated by defining boundary conditions of patients in the clinical results. Two sample which were taken were, a stenosed and a normal artery of eleven years old patients and was explored by pressure and wall shear stress distributions. In different artery profiles, wall shear stress, pressure and velocity results were siginificantly different. However, in the crititcal regions, the arterial wall was in the risk of deformation after narrowness in the lumen. Modeling and simulating depicted evident information about mechanical effects of blood flow. Understanding the flow regions and effects in different regions were, the most effective and productive treatments which were particular for the patient and could be chosen by realistic modeling and simulation. However, comparing biomaterials, examining tools and developing better designs will be the goal for medical researches and for the benefits of the patients.
Alexei A Yeliseev
National Institute on Alcoholism and Alcohol Abuse USA
Title: Human cannabinoid receptor CB2: Expression, functional and structural studies
Biography:
Alexei Yeliseev received his Ph.D. in biochemistry in 1987 from the Russian Academy of Sciences in Moscow and did his postdoctoral research in enzymology in Marburg, Germany and in molecular biology and biochemistry in Cambridge, UK. After moving to US in1993 he worked as a Research Fellow at the University of Texas Medical School at Houston and as a Senior Scientist at Hoffmann-La Roche and later, at Kosan Biosciences, Inc. He later moved to the National Institutes of Health where he currently heads the protein biochemistry group developing technologies for expression, purification, and analysis of recombinant G protein-coupled receptors. In addition to his research work he serves as a member of editorial board of Protein Expression and Purification and Journal of Receptor, Ligand and Channel Research
Abstract:
The human cannabinoid receptor CB2 belongs to the class A of hepta helical G protein-coupled receptors (GPCR) and is an attractive target for the development of drugs for management of pain, inflammation, osteoporosis and treatment of immunological disorders. High resolution structural studies are critical to obtain insights into the molecular mechanisms of ligand binding and activation of CB2. We developed methods for expression in milligram quantities, purification, reconstitution in lipid bilayers and stabilization of the functional recombinant CB2 as well as efficient stable isotope labeling of CB2 by high density fermentation were developed enabling NMR studies. NMR analysis of labeled receptor reconstituted in proteoliposomes in agonist or inverse-agonist-bound form will be reported. Monoclonal antibodies were raised against the purified CB2 and the affinity of interaction with CB2 was determined by surface plasmon resonance. Finally, we demonstrate the specific effects of lipids with negatively charged head group in activation of CB2 reconstituted in proteoliposomes as measured by an in vitro G protein activation which may have important physiological significance as manifested in natural membranes of various lipid compositions.
Lekshmi R Nath
Rajiv Gandhi Centre for Biotechnology, India
Title: 4-O-Methylgallic acid isolated from the leaves of the mistletoe growing on Saraca asoca induces caspase-mediated apoptosis in cervical cancer cells via down-regulation of HPV
Biography:
Lekshmi R Nath obtained her MPharm from K L E Society’s college of Pharmacy. Currently she is pursuing her PhD in Biotechnology at Rajiv Gandhi Centre for Biotechnology, Kerala, India. She has published more than 5 papers in reputed journal.
Abstract:
Cervical cancer, the second most common gynecological malignancy remains a leading cause of cancer death in women in developing countries. The major etiologic factor for cervical cancer is high-risk HPV infection. Several studies indicate that the E6 and E7 gene products play a critical role in cervical carcinogenesis. Medicinal plants have gained much importance in the development of new cancer treatment strategies and extracts derived from mistletoe have been shown to kill cancer cells in vitro. We have isolated 4-O-Methylgallicacid (4-OMGA) from the mistletoe growing on Saraca asoca which induces apoptosis in cervical cancer cells. Among various cervical cancer cells screened the compound exhibited maximum cytotoxicity in HeLa (IC50-17 µg/ml) followed by SiHa (IC50-37 µg/ml) both of which are HPV positive cells while the cytotoxicity was comparatively less in the HPV negative cell line C33A (IC50-91 µg/ml). The compound induces apoptosis is HeLa as evidenced by caspase activation and PARP cleavage. We also observed that the compound is down-regulating the expression of E6 /E7 (both RNA and protein level) in HeLa cells in a dose dependent manner.
- Nanobiotechnology
Location: Flemings Conference Hotel
Session Introduction
Arshad Islam
Universidade Federal de Minas Gerais, Brazil
Title: Nanosystems formed by amphiphilic antimony(v) complexes incorporating amphotericin B for the treatment leishmaniasis
Time : 11:30-11:55
Biography:
Arshad Islam has completed his PhD from Universidade Federal de Minas Gerais, Brazil and currently working as Post-doctoral researcher at Laboratory of Biophysics and Nanostructured Systems, Department of Physiology and Biophysics, Institute of Biological Sceinces, Universidade Federal de Minas Gerais. He has published 4 papers in reputed journals
Abstract:
This work aimed to developan oral formulation of amphotericin B (AmB) for treatment of leishmaniasis. AmB is an antifungal and antibacterial macrolide polyenederived from Streptomyces nodosus strain, that belongs to the group of second generation anti-leishmanial drugs and is extensively used in case of failures in the treatment with antimonial compounds. AmB was incorporated into nanosystems formed by amphiphilic antimony(V) complexes with ligands of alkylmethylglucamide series (L8 and L10, with 8 and 10 carbon chain, respectively). Incorporation rate of 0.2% AmB into SbL8 and SbL10 dispersions was determined using an HPLC-based technique and was found to be 84±1% and 74±1%, respectively. The characterizarion of SbL10-AmB and SbL8-AmB by circular dichroism and UV-visible spectroscopies showed that AmB is present predominantly under the monomeric form in both SbL8 and Sb10 nanosystems, which is the least toxic form to the host and potentially most bioavailable. The potential for the oral treatment of visceral (VL) and cutaneous leishmaniasis (CL) was evaluated in murine models in comparison to the standard drug Anforicin B® or Glucantime® administered intraperitoneally or orally. In Balb/c mice infected with Leishmania amazonensis, the SbL10-AmB mixed formulation (170 mgSb/kg and 14 mg AmB/kg, each 2 days by oral route) resulted in a significant decrease of the lesion size, when compared to orally administetred Glucantime® and SbL10 (170 mg Sb/kg, each 2 days), Anforicin B® (>1mg/kg/each 5th day, by intraperitoneal route) and control saline group. In Balb/c mice infected with Leishmania infantum, both the SbL10-AmB and SbL8-AmB mixed formulations given orally (170 mg Sb/kg and 14 mg AmB/kg per day) reduced significantly the parasite load in the liver compared to the untreated control, to a similar level as AmB given intraperitoneally (0.9 mg/kg/day). This study established for the first time the potential of mixed SbL10-AmB and SbL8-AmB formulations for the oral treatment of both cutaneous and visceral leishmaniasis , indicating their potential for further development and applications.
Cletus A Ukwubile
Federal Polytechnic Bali, Nigeria
Title: Anti-microbial properties and characterization of annona muricata linn loaded chitosan nanoparticles against cancer associated microbes
Biography:
Abstract:
Annona muricata chitosan nanoparticles (AMCS), nano-seized drug carriers were prepared by ionic gelation of chitosan and sodium tripolyphosphate (TPP) anions. Antimicrobial properties and characterization of A. muricata chitosan formulation of leaf methanol extract were evaluated on ten cancer associated microbes; Bacillus subtilis NCTC8239, Salmonella typhi ATCC9184, Staphylococcus aureus NCTC6571, E.coli NCTC10418, Staphylococcus aureus (clinical isolate), MRSA (clinical isolate), Streptococcus pyogenes (clinical isolate), Klebsiella pneumonia ATCC 10031, Candida albican ATCC 19231 and Staphylococcus aureus NCTC 6571 using agar well diffusion method. AMCS exhibited great antimicrobial properties against all the microbes which were significantly different from that of the broad spectrum antibiotic ceftriaxone 10 mg; USP. Compatibility of A. muricata loaded chitosan nanoparticles as revealed by atomic force and transmission electron microscopes showed that chitosan nanoparticles are effective drug carriers for both antimicrobial and anticancer agents. The study showed that chitosan loaded plant extract of Annona muricata is a sure promise towards treating some infectious diseases such as cancer and microbial infections.
Sandeep Pandey
Awadhesh Pratap Singh University, India
Title: Dynamics of nanotechnology tools to combat plant pathogens
Biography:
Dr. Pandey started his career as a Lecturer in colleges in Mumbai University. He persuaded his Doctorate in Environmental Biology from A.P.S. University, Rewa in 2005. He joined the same university in 2005 and is working in the Department of Biotechnology, Environmental Sciences, Botany and Forestry. He is specialized in Plant Pathology. His area of research also includes environmental biology, ethnobotany, forestry, environment management, bioinformatics and nanotech sciences.
Abstract:
Nanotechnology has emerged with varied scopes in the field of plant pathology. In recent year the science of plant pathology has witnessed major breakthroughs in plant defense and disease combating programs. The nanotubes, an important tool of nanotechnology which is a cylindrically shaped carbon molecules having a diameter of about one nanometer can be exploited to deliver proteins, nucleic acids and drugs to cells. This technique also helps to recognize and fight pathogens in agriculture crops. Although carbon nanotubes had toxic effects because they induce programmed cell death in plant cells but do not show any adverse impact at the tissue level indicating that injecting cells with carbon nanotubes caused only limited injury. Thus to design a disease combating nanotechnology model (DCNM) requires assessment of bioavailability and toxicity of carbon nanotubes and their impurities on the crops. The crops should also be tested for their cell viability, genetic material, gene expression, seed germination, seed growth and various other physiological, biochemical and genetic characteristics before applying the nanotechnology principles. The doses of nanotubes can act as a limiting factor in these programs. The use of magnetic nanoparticles through magnetic field gradients has been proved significant in selected plant tissues. The magnet helps these nanoparticles that are charged with different substances to get introduced within the plants and if necessary concentrated into localized areas modifying pathogenesis mechanism. The safer use of nanotechnology techniques to combat agriculture crop pathogens and evaluation of new tools for a successful plant disease management programs are reviewed.
Eka Sunarwidhi Prasedya
Fukushima Medical University, Japan
Title: Sulphated polysaccharides from seaweeds has potential to inhibit cell growth and cell cycle arrest of human cancer cells
Time : 11:55-12:20
Biography:
Eka Sunarwidhi Prasedya is a PhD candidate pursuing his PhD in Fukushima Medical University, Japan. Previously, he earned his Bachleor’s degree from Gadjah Mada University, Jogjakarta, Indonesia and Master’s degree from Universiti Malaya, Kuala Lumpur, Malaysia.
Abstract:
Sulphated polysaccharides from marine algaes has been previously reported to have therapeutic properties. Esp ecially, sulphated polysaccharides such as carrageenaand n fucoidan are known to exhibit strong antitumor activities. However, little is known about the mechanism of fucoidan or carrageenan an titumor activities towards human cells. In this study, firstly, we evaluated the effects of fucoidan and carrageenan towards cell growth of HeLa cells. HeLa cells were cultured with various concentrations of fucoidan (0-100 µg/mL) and carrageenan (0-1000 µg/mL). Both sulphated polysaccharides were seen to i nhibit cell growth of HeL a cells. To confirm whether cell growth is truely inhibited or not, fucci fluorescent ubiquitiniation-based cell cycle indicator were used to evaluate cell cycle arrest due to fucoidan and carrageenan treatment. Cell cycle was seen to be arrested at the G1 phase which indicates that later on cells would undergo apoptosis. The findings of this study so far gives the idea that fucoidan and carrageenan inhibits cell growth of HeLa cells by arresting cell cycle. Future studies are still needed to provide better understanding of the antitumor mechanisms of marine algae sulphated polysaccharides.
Akbar Vaseghi
Islamic Azad University, Iran
Title: Gold nanoparticles for biomedical applications
Time : 12:20-12:45
Biography:
Akbar Vaseghi has completed his master at the age of 27 years from Tarbiyat Modares University. He has published more than 10 papers in reputed journals. He has been working as a research assistant in the Nanobiosensor particular Drug Design, Imaging for cancer cells whit using Gold Nanoprobes by possibility attach in biological targets (DNA and peptide).
Nanobiotechnology laboratory and Computational Nano- design. He has experience in this field whit investigation, especially by researchAbstract:
surface chemistry be produced q uickly and inexpensively; also with their unique optical properties have been employed for detection of a variety of pathogenic microorganism including bacteria, viruses and fungi, there are differed ways for synthesize GNPs which are widely employed. An important challenge in medical diagnostics is to design all-in-one contrast agents that can be qualitative or quantitative detected with multiple techniques such as Magnetic Resonance Imaging (MRI), X-ray Computed Tomography (CT), Photothermal Therapy (PTT), Photodynamic Therapy (PDT), Positron Emission Tomography (PET), Single Photon Emission Tomography (SP ECT) or ultra-efficient Fluorescence Quenchers (FI) and Quantum Dots (QDs). Gold nanoprobes have gained significance as novel pharmaceutical compounds to be used for imaging purposes and targeted delivery of therapeutic cargoes into cancer cells. Taken together, special features of nanoprobes and their diverse range of applications highlight their importance as valuable diagnostic and therapeutic tools. We made attempt to design next generation nano-chips through using gold nano-probes particularly gold nanoparticles and nano-rods for detection of pathogens and cancer cells. To this end, we attached bio-barcodes to gold nanoprobes to achieve detection and therapeutic objectives. However, due to the explosion of publications about applications of AuNPs, this current review we describe a method to quantify recent research advances of gold nano-. probe
Nanomaterials particularly Gold Nanoparticles (GNPs) features unique physicochemical properties so that represent enormous promise for diagnosis and treatment of human diseases. These particles have a biocompatibility, size, provide a high surface-to-volume ratio and ease for fictionalization or characterization, their low toxicity, high extinction coefficients, straight forward synthesis methods as well as an extensive knowle dge of theirNasirzadeh Keyvan
Imam Hossein Comprehensive University, Iran
Title: Preparation and characterization of alginate beads reinforced with silica nanoparticles for microbial cell culture
Time : 12:45-13:10
Biography:
Abstract:
Silica-alginate nanocomposites were prepared by reinforcing the alginate beads with sili ca nanoparticles and used for microbial cell culture. Nonporous silica nanoparticl es (NSNs) and mesoporous silica nanoparticles (MSNs) were prepared and then amine-functionalized with N-(2-aminoethyl)-3-aminopropyl trimethoxy-silane (EDS) to prepare positively charged amine-n. Different concentration of the prepared unfunctionalized and amine-functionalized silica nanoparticles were utilized to prepare reinforced alginate beads. Mechanical stability of the prepared beads was investigated and the Young's modulus values were calculated. DH5α cell were entrapped in the prepared beads and then their growth was studied. In addition, the toxicity of the silica particles on the growth of the cells was also studied. Young's modulus values of the beads reinforced with all the silica nanoparticles were markedly improved in comparison to that of the beads without any silica anoparticles nanoparticles . The Young's modulus values of the beads reinforced with amine-functionalized MSNs was measured to be app roximately 4.5 folds higher than that for the bare alginate beads. The existence of silica nanoparticles in the alginate beads not only increases their mechanical stability but also decreases the leakage of cells from the beads. In addition, the results showed that the various silica nanoparticles prepared in this work does not show any significant effects on viability of the encapsulated cells. Finally, the superior properties of the prepared nanocomposite beads recommend them as a suitable cell culture system for microbial cells' cultivation for various applications.
F Kashanian
University of Tehran, Iran
Title: A novel method to effectively remove waste metabolic products
Biography:
Faezeh kashanian is a PhD student of nano-biotechnology at Tehran University from the year of 2012. She has been working in the Biotech Laboratory of Magnetic Nanoparticles for 3 years. During all these past 10 years, she has taught Fundamentals of physics, biophysics and Quantum physics. Master of Science was solid state of physics and Bachelor of Science was in Applied Physics and Software computer.
Abstract:
The kidney is responsible for regulating acid base balanced and electrolyte concentration example sodium and chloride as well as excreting waste products produced by metabolism like urea and creatinine. It is known that these uremic solutes retention indicating as pathophysiological signs and symptoms lead to kidney insufficiency. In the acute renal failure to protect the health of the patient although dialysis treatment is used to remove mentioned waste products and regulating intracellular fluids but it has some disadvantages. Here introduced Nano Magnetic Bio Polymers Systems (NMBPS) for smart removal of metabolic waste products in blood by means of a kind of absorbents is based on molecular imprinted method. Highly selective separation of metabolites of blood by taking the advantage of applying in the new dialysis device for removing the metabolites is novel and efficient therefore, designing a magnetic capturing mechanism to get rid of all foreign material (magnetic nanoparticles) before return the blood to the patient body is necessary. Prepared NMBPS possessed easy accessibility to the targeted sites and showed fast kinetics, high capacity and favorable selectivity. Successful application of this method in the capturing of the wastes from blood and acceptable recovery after a reasonably mild elution suggested that NMBPS can be used for selectively capturing metabolites in the blood. The benefits of this separation method are capturing the high molecular weight toxins, lower losing the Blood proteins example Albumin and lower long term side effects.
- Environmental Biotechnology
Location: Flemings Conference Hotel
Session Introduction
Shree Kumar Apte
Bhabha Atomic Research Centre, India
Title: Genetic engineering of Deinococcus radiodurans for uranium bioremediation from high radiation environment
Time : 15:05-15:30
Biography:
Shree Kumar Apte, a Former Director of Bio-Science Group, BARC, is currently Emeritus Professor at the Homi Bhabha National Institute, Mumbai, India. His laboratory has extensively studied stress and adaptive responses in bacteria and plants in response to agricultural stresses and ionizing radiation and developed eco-friendly biotechnologies for agricultural and environmental applications. He is an elected fellow of all the national science academies and agriculture academy in India and has over 170 research publications in high impact international journals to his credit
Abstract:
In nature, uranium occurs over a wide range of concentrations and is generally toxic to all living cells. Exploitation of uranium by the nuclear industry generates acid/alkaline waste, wherein uranium is found at low (<1-2mM) concentration. Removal of even such low concentrations of uranium is desirable for safe disposal of the waste, but is difficult to achieve by physico-chemical methods. Bioremediation, especially bio-precipitation as uranyl phosphate, is an efficient way to remove uranium from such waste, where high levels radiations also prevail. The radio-resistant microbe, Deinococcus radodurans, was genetically manipulated to individually over-express acid and alkaline phosphatases using deinococcal strong promoters, including the radiation-induced Pssb promoter. Lyophilization was successfully employed to preserve both the phosphatase activities and uranium precipitation ability of recombinant cells up to 1 year at ambient temperature. Such cells could remove 7-11 g U/g dry weight of the biomass
Gabriela Briceno
Universidad de La Frontera, Chile
Title: Selection of an actinobacteria consortium for enhancing diazinon degradation
Time : 15:50-16:15
Biography:
Gabriela Briceno Munoz has completed her PhD in Natural Resource Sciences at the age of 31 years from Universidad de La Frontera (Chile). She has developed research in environmental biotechnology like studies of pollutants degradation by actinobacteria and environmental fate of pesticides. Currently, she is a Researcher in the Scientific and Technological Bioresource Nucleus and Professor of the Department of Chemical Science and Natural Resources. She is author of chapters of books, several scientific publications and regularly is working as Referee for scientific journals
Abstract:
The biological treatment of pesticides constitutes a promising alternative for a safe, efficient and economical elimination of pesticides. In this way, biological treatments for removing organophosphorus pesticides, toxic and worldwide used pesticides from contaminated matrices are needed. The aim of this work was to select an actinobacteria consortium to enhance the diazinon degradation. For this, organophosphorus-degrading actinobacteria identified as Streptomyces sp. strains AC5, AC6, AC7, AC9, GA3, GA11, ISP4 and ISP13 were used. In 30 mL of minimal medium containing 50 mg L-1 diazinon as only carbon source, single strains of actinobacteria were added. After 96 hours of incubation, microbial growth, intracellular protein content, protein profile and the concentration of diazinon and their metabolite 2-isopropyl-6-methyl-4-pyrimidinol (IMHP) were analyzed. The results showed that biomass and protein content increased with the diazinon addition. Thus, some of the actinobacteria showed prominent bands of proteins enhanced in response to diazinon application. About 10-30% of diazinon degradation for single actinobacteria was observed and only three strains showed IMHP production. However, when mixed cultures of two, three, four and five actinobacteria strains were evaluated, the diazinon degradation was increased reaching values close to 90%. The quadruple culture composed by the strains AC5, AC9, GA11 and ISP13 presented the best diazinon degradation which occurs during the first 72 hours with a decrease of IMHP over time. Therefore, we conclude that the selected actinobacteria consortium is a promising alternative to increase the diazinon degradation.
Heidi Schalchli
Universidad de La Frontera, Chile
Title: Natural products of Anthracophyllum discolor: Ligninolytic enzymes and antifungal volatile compounds
Time : 16:15-16:40
Biography:
Heidi Schalchli has completed her PhD from Universidad de La Frontera. She is a Postdoctoral Researcher at the Scientific and Technological Bioresource Nucleus (BIOREN) and Teacher at the Chemical Engineering Department, Universidad de La Frontera
Abstract:
White-rot fungi play important roles in ecosystems mainly because of their extracellular enzymatic system and their production of chlorinated aromatic compounds that act as decomposers of organic matter, antibiotics for protecting fungi, methyl donors and/or substrates for H2O2-generating oxidases. In this study, we evaluated the production of ligninolytic enzymes and antifungal volatile organic compounds (VOCs) by A. discolor Sp4 using Potato Peels (PP) and Discarded Potato (DP) as nutritional support. The manganese-dependent peroxidase (MnP) was evaluated by monit oring the oxidation of 2, 6-dimethoxyphenol. Beside the production of MnP, the discoloration of remazol brilliant blue R (RBBR) was also determined using a qualitative assay. The antifungal activity of VOCs against Mucor miehei and Fusarium oxysporum was evaluated using a bi-compartmented plate assay. Finally, VOCs released from mycelial cultures were analyzed by headspace solid phase micro-extraction and gas chromatography mass spectrometry. The highest MnP and MiP activities (163 U L-1 and 24 U L-1) were obtained at day 15 of incubation and a complete RBBR discoloration was observed. Although both potato wastes supported the ligninolytic activity, a higher MnP activity was obtained using PP than DP. The A. discolor volatiles inhibited approximately 62% and 76% the mycelial growth of M. miehei on PP agar and DP agar media, respectively. Nevertheless, the plant pathogen F. oxysporum was slightly inhibited (approximately 10%). The major VOCs detected were chlorinated aromatic compounds (over 50% relative area). The obtained natural products have multiple biotechnological applications among which are pollutant degradation and plant protection.
Sandra Borkowska-Heurtaux
Glasgow Caledonian University, Scotland
Title: The influence of growth conditions (temperature and oxygen supply) on manganese (II) sorption by viable and autoclaved Lactococcus lactis var. lactis NCIMB 6681
Time : 16:40-17:05
Biography:
Sandra Borkowska-Heurtaux completed her BSc (Hons) and MPhil and is a final year PhD student at Glasgow Caledonian University. Currently, she is working on biosorption of manganese and zinc by Lactococcus lactis var. lactis, Natrinema pallidum, Hydrodictyon reticulatum and Cladophora glomerata using microbiology and analytical chemistry methods. Prior to beginning the PhD program, she worked as a Research Associate at Edinburgh Napier University. Her work focused on microbial testing to prove efficacy of advanced disinfectant against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Enterobacter cloacae and Salmonella typhimurium. Her areas of interest include microbiology, biotechnology, genetics and molecular biology.
Abstract:
Manganese (II) is an essential element required for normal growth and development of humans, animals and plants, however it has a tendency to accumulate in some organisms, what leads to higher, potentially toxic, level within the food chain. Mn2+ sorption properties of Lactococcus lactis var. lactis, a non-pathogenic bacterium widely used in the dairy industry were studied as a function of four growth conditions: Cells were cultivated aerobically and with reduced oxygen at 30° C and 37° C. Additionally, biosorption properties of live and autoclaved cells were compared. L. lactis showed very competitive capability to sorb Mn2+ over 5 days and pH drifts in the experimental suspensions demonstrated an involvement of ion exchange mechanisms in Mn2+ sorption. Viability of L. lactis during sorption experiments was studied by serial dilutions and plate count methods with the biggest decrease in a cell numbers observed at 24 and 72 hours contact time. Sorption capacity of live L. lactis cultivated under four different conditions towards Mn (II) ranged 34-50 mg/gdw. Autoclaved biomass showed much lower sorption capacity (20-39 mg/gdw) but this range is among the highest removal capacities towards Mn2+ seen in previous studies using various (non-living) biomasses. The obtained results are the first report showing Mn2+ sorption by viable and autoclaved cells of L. lactis as a function of different growth conditions and metal loadings. It is also among the first work investigating the difference between viable and dead microbial cells.
Hela Ben Amor-Ben Ayed
Centrale Supelec Paris, France
Title: Biosorption of magnesium in Chlorella vulgaris under different culture conditions
Time : 17:05-17:30
Biography:
Hela Ben Amor-Ben Ayed is pursuing her PhD. She obtained an Engineering Diploma in Biology from the National School of Engineering of Sfax, Tunisia which she followed with a Master’s on Environmental Biotechnology from the Institute of Biotechnology of Sfax. During her Master’s degree, she worked on potable water treatment in Paris. She has submitted 3 manuscripts during her PhD in reputed journals, the first one of which has been published
Abstract:
A study investigating the accumulation of magnesium by Chlorella vulgaris under different culture conditions is described. The dissolved and biomass-associated concentrations of magnesium were measured with atomic absorption spectroscopy during the course of C. vulgaris growth under autotrophic or mixotrophic conditions both in shake-flask (100 mL) and photo-bioreactor (5 L) cultures. The adsorbed (extracellular) and absorbed (intracellular) ions associated with the biomass were determined using adapted published methods. During the experiments, a clear relationship between the growth extent of C. vulgaris and magnesium removal from the medium was observed. In an autotrophic shake-flask culture with a medium concentration of 19.1 mg Mg2+/L, 78% of the initial magnesium content of the medium was associated with the biomass, of which 6% was adsorbed on the cell wall and 72% absorbed into cells at the end of the experiment (480 hours). In a mixotrophic photo-bioreactor culture with glucose (10%) as the sole organic carbon source, C. vulgaris accumulate 90% of the initial magnesium content of the growth medium, of which 4% was adsorbed on the cell wall and 86% absorbed by the biomass. Magnesium association with C. vulgaris was faster and more extensive under mixotrophic conditions. These results could be interesting for the accumulation of metal ions by microalgae on an industrial scale.