Scientific Program

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Scientific Program Day 1
Scientific Program Day 2

Day 2 :

Keynote Forum

Igor l Katkov

Belgorod National State Research University, Russian Federation

Keynote: Stopping biological time: Science and art of biostabilization

Time : 10:00-10:40

Conference Series Euro Biotechnology 2018 International Conference Keynote Speaker Igor l Katkov photo
Biography:

Igor L Katkov is a trained biophysicist with 30+ years of experience in cryobiology and cryogenic engineering. His last years of research have been focused on the fundamental aspects of kinetic vitrification (K-VF) as well on designing the practical system for K-VF KrioBlast™ (in cooperation with V F Bolyukh). Currently, the Head of the Laboratory of the Amorphous state at the Belgorod National Research University BelSU, Russia. He has recently accepted a
Professor level position as the Head of the Laboratory of Cryobiology at the V I Kulakov Research Center of Obstetrics, Gynecology and Perinatology (RCGOP), Moscow, Russia and Chief Scientific Officer of Celltronix, San Diego, CA, USA.

Abstract:

Biostabilization (a.k.a. biopreservation) is a process that leads to cessation of the basic chemical and biological reactions
so the biosamples can be pooled and stored (biobanked) for long time. There are 5 basics ways of achieving long-term
storage, which ALL essentially lead to vitrification of cells, namely: slow freezing (SF), equilibrium vitrification (E-VF), kinetic
vitrification (K-VF), freeze-drying (lyophilization) and vacuum/air flow drying at temperatures above 0°C (xeropreservation).
Different combinations of the 5 basic biopreservation technologies such a preliminary drying before cryogenic slow freezing
or vitrification is also possible. Author will discuss a phase diagram that shows all 5 basic ways of biostabilization and will
discuss pros and cons of all approaches. A special emphasis will be put on the kinetic vitrification as it does not require the
high concentrations of (or does not need at all) potentially toxic and osmotically damaging exogenous permeable intracellular
vitrificants (also called cryoprotectants). Author will also present KrioBlast-2, a pilot version of the KrioBlast™ platform for
cryopreservation by K-VF. Preliminary experiments on K-VF of human pluripotent stem cells and spermatozoa, which
showed an equally excellent (80-90% of the untreated control) will be also discussed. A more advanced version KrioBlast-3
will be discussed in the concurrent presentation.

Keynote Forum

Sergey Suchkov

I M Sechenov First Moscow State Medical University, Russia

Keynote: Antibodies with functionality as a new generation of translational tools to monitor, to predict and to prevent demyelination

Time : 10:00-10:40

Conference Series Euro Biotechnology 2018 International Conference Keynote Speaker Sergey Suchkov photo
Biography:

Sergey Suchkov graduated from Astrakhan State Medical University and was awarded with MD and maintained his PhD and Doctor’s degree. He was working for Helmholtz Eye Research Institute and Moscow Regional Clinical Research Institute. He was a Secretary-in-Chief of the Editorial Board, Biomedical Science, an international journal published jointly by the USSR Academy of Sciences and the Royal Society of Chemistry, UK. Currently, he is a Director of Center for Personalized Medicine, Sechenov University; Chair of the Department for Translational Medicine, Moscow Engineering Physics University and Secretary General of United Cultural Convention, Cambridge, UK. He is a Member of the New York Academy of Sciences; American Chemical Society; American Heart Association; AMEE, Dundee, UK; EPMA, Brussels, EU; PMC, Washington, DC, USA and ISPM, Tokyo, Japan.

Abstract:

Abs against myelin basic protein/MBP endowing with proteolytic activity (Ab-proteases with functionality) is of great value to monitor demyelination to illustrate the evolution of multiple sclerosis (MS). Anti-MBP auto-Abs from MS patients and mice with EAE exhibited specific proteolytic cleavage of MBP which, in turn, markedly differed between: MS patients and healthy controls; different clinical MS courses and; EDSS scales of demyelination to correlate with the disability of MS patients to predict the transformation prior to changes of the clinical course. Ab-mediated proteolysis of MBP was shown to be sequence-specific whilst demonstrating five sites of preferential proteolysis to be located within the immunodominant regions of MBP and to fall inside into 5 sequences fixed. Some of the latter (with the highest encephalitogenic properties) were proved to act as a specific inducer of EAE and to be attacked by the MBP-targeted Ab-proteases in MS patients with the most severe (progradient) clinical courses. The other ones whilst being less immunogenic happened to be EAE inducers very rare but were shown to be attacked by Ab-proteases in MS patients with moderate (remission-type) clinical courses. The activity of Ab-proteases was first registered at the subclinical stages 1-2 years prior to the clinical illness. About 24% of the direct MS-related relatives were seropositive for low-active Ab-proteases from which 22% of the seropositive relatives established were being monitored for 2 years whilst demonstrating a stable growth of the Ab-associated proteolytic activity. Moreover, some of the low-active Ab-proteases in persons at MS-related risks (at subclinical stages of MS), and primary clinical and MRT manifestations observed were coincided with the activity to have its mid-level reached. Registration in the evolution of highly immunogenic Ab-proteases would illustrate either risks of transformation of subclinical stages into clinical ones, or risks of exacerbations to develop. The activity of Ab-proteases in combination with the sequence-specificity would confirm a high subclinical and predictive (translational) value of the tools as applicable for personalized monitoring protocols. Ab-proteases can be programmed and re-programmed to suit the needs of the body metabolism or could be designed for the development of principally new catalysts with no natural counterparts. Further studies on targeted Abmediated proteolysis may provide a translational tool for predicting demyelination and thus the disability of the MS patients.

Keynote Forum

Igor l Katkov

Belgorod National State Research University, Russian Federation

Keynote: KrioBlastTM-3 - a three module system for efficient cryopreservation of unfreezable cells
Conference Series Euro Biotechnology 2018 International Conference Keynote Speaker Igor l Katkov photo
Biography:

Igor L Katkov is a trained biophysicist with 30+ years of experience in cryobiology and cryogenic engineering. His last years of research have been focused on the fundamental aspects of kinetic vitrification (K-VF) as well on designing the practical system for K-VF KrioBlast™ (in cooperation with V F Bolyukh). Currently, the Head of the Laboratory of the Amorphous state at the Belgorod National Research University BelSU, Russia. He has recently accepted a Professor level position as the Head of the Laboratory of Cryobiology at the V I Kulakov Research Center of Obstetrics, Gynecology and Perinatology (RCGOP), Moscow, Russia and Chief Scientific Officer of Celltronix, San Diego, CA, USA.

Abstract:

As we have stated before, there are 5 basics ways of achieving long-term storage, which ALL essentially lead to vitrification of cells, namely: slow freezing (SF), equilibrium vitrification (E-VF), kinetic vitrification (K-VF), freeze-drying (lyophilization), and va San Diego vacuum/air flow drying at temperatures above 0oC (xeropreservation). Previously, we presented KrioBlast-2, a pilot version of the KrioBlast™ platform for cryopreservation by kinetic (very fast) vitrification. One of the major advantages of K-VF over the existing approach for vitrification (E-VF) is that K-VF does not need the high concentrations of potentially toxic and intracellular vitrificants (also called: cryoprotectants, which is not exactly correct in this case) such as DMSO, ethylene glycol, dimethyl sulfamide. The pilot experiments on human pluripotent stem cells and spermatozoa, which showed an equally excellent (80-90% of the untreated control), were presented. The other key advantage of K-VF is its universality so the system is equally suitable for any kind of cells and tissues as soon as the characteristic thermal time of the system, which basically depends on the geometry of the cryo container with the sample, is sufficiently short. In this presentation, we will present the future development, the industrial three module system KrioBlast-3 that comprises 1) the cooling chamber for hyperfast cooling, 2) the intermediate module for shipment or long term storage in liquid nitrogen, and 3) the rewarming module. The second module has two port sites for the cooling and the rewarming modules so the system resembles a space station. All operations of cooling, storage/shipment, and warming are done without any contact of the sample with the ambient environment. The specific cryo containers for K-VF, namely VitriPlateTM, VitriCombTM, and VitriScanTM for vitrification of cells in suspension, packed in straws, and attached to surface in multiwell systems respectively are also discussed.

  • Environmental Biotechnology | Bioprocess and Fermentation Technology | Agriculture Biotechnology Molecular Biotechnology and Genetics | Pharmaceutical BioteEnvironmental Biotechnology | Bioprocess and Fermentation Technology | Agriculture Biotechnology Molecular Biotechnology and Genetics | Pharmaceutical Biotechnology and Drug Designchnology and Drug Design
Location: Winter garden
Speaker

Chair

Igor Katkov

Belgorod National State Research University, Russian Federation

Speaker

Co-Chair

Farouk El-Baz

National Research Centre, Egypt

Session Introduction

Farouk K El-Baz

National Research Centre, Egypt

Title: Growing scenedesmus obliquus microalgae in photobioreactor for biomass and biodiesel production
Speaker
Biography:

Farouk K El-Baz has completed his PhD from Cairo University, Cairo. He is a Professor of Biochemistry, the Principal Investigator of biodiesel production from algae as a renewable energy source project which funded by EU. He is also the PI of Industrial Pharmaceutical Alliance (NRC) sponsored by the Academy of Scientific Research and Technology, Egypt. He is the Director of Algal Technology Unit/ NRC, Cairo, Egypt. He has published 152 papers in international journals; He has supervised 18 theses, and serving as the Reviewer of many international journals.

Abstract:

The aim of this study is to investigate the biomass and oil production capacity of S. obliquus grown in outdoor photobioreactors (PBR) under nitrogen repletion and starvation conditions. The volume of PBR was 4000 liters as a demo unit with the facility of computerized controlled system. The results showed that, the maximum biomass achieved with the highest dilution rate (0.25%) was 43.2 gm-2d-1. This was detected when the dry weight was 2.1 g/L. The maximum oil content reached to 26%±0.23 after 29 days under N repletion. However under nitrogen starvation, the oil content was dramatically increased and reached to 41.9%±0.6 after 8 days. Fatty acids profile showed that, both saturated and unsaturated acids were detected. The major saturated fatty acids were palmitic and stearic acids. The unsaturated fractions were detected as palmitoleic, oleic, linoleic and linolenic acids. The fatty acids with four or more double bonds were not detected. Total saturated fatty acids represented 60.47% and 67.43% under nitrogen repletion and nitrogen starvation respectively. The use of photobioreactor for the production of algae is economically feasible, where there is a large amount of sun energy available, which provides a great saving for energy. A high quality of biodiesel could be produced from microalgae S. obliquus and used efficiently and environmentally safe in conventional diesel engine

Monika Cserjan

University of Natural Resources and Life Sciences, Austria

Title: 12:40-13:00
Speaker
Biography:

Monika Cserjan has completed her PhD at the University of Natural Resources and Life Sciences, Vienna in 1998. She is Senior Scientist in the Christian Doppler Laboratory for production of next-level biopharmaceuticals in E. coli at the Department of Biotechnology (Fermentation Technology Group), Vienna and Project Leader at the Austrian Centre of Industrial Biotechnology (ACIB).

Abstract:

Although E. coli is the most prominent bacterial production host for recombinant proteins, some proteins with high economic potential can still hardly be produced at remunerative levels. We selected four different Fabs (Fragment, antigen binding) (BIBH1, BIWA4, CIMZIA and FabX) with identical constant domains representing such challenging proteins. Fab yield can be affected by miss-folding, aggregation or unbalanced expression, translation and translocation levels of sub-units, making it still challenging to efficiently design expression systems and production processes. For translocation to the periplasm a post-translational (ompA) and a co-translational (dsbA) leader sequence were used. E. coli BL21(DE3) and E. coli HMS174(DE3) were transformed either via pET vectors or genome integration. The resulting 32 clones, were cultivated under fed-batch like conditions in the BioLector. Cell growth was not affected by leader/Fab combinations but yield of correctly folded Fab ranged from 0 to 12.5 mg/g CDM. Higher expression rates caused higher amounts of free light chain and K12 strain reached higher yields. Except of CIMZIA with the dsbA leader, genome integrated versions showed higher Fab yields, reduced levels of free light chain and basal expression than plasmid-based systems. Independent from used expression system, highest yields were obtained with CIMZIA followed by BIWA4, BIBH1 and FabX. Leader sequence cleavage-efficiency for DsbA was significantly lower than for OmpA, both showed lowest with CIMZIA. Summarizing, we showed that the selected set of host/gene dosage/leader/Fab combinations resulted in a broad range of variation in terms of Fab yields and processing and will be studied detailed in bench-scale fermentations. 

Petrova Natalia Vladimirovna

Institute of General Pathology and Pathophysiology, Russia

Title: A new direction for viral infections management
Speaker
Biography:

Petrova Natalia Vladimirovna has graduated from Saratov State University named after Chernyshevsky and since that time she launched into the scientific work. She was honoured to take part in international scientific project Marie Curie Actions: International Research Staff Exchange Scheme (Institutte of Transfusion Medicine, Berlin). At the moment, she is involved in her Postdoctoral studies at the Institute of General Pathology and Pathophysiology, where she is currently taking a position of Research Associate. She is a co-author of seven papers in reputed journals.

Abstract:

A New Direction For Viral Infections Management: Viral diseases are still one of the most pressing global problems of society throughout the world. The problem of effectiveness and safety for currently used antiviral therapy is still a major concern; a principally new approaches and the creation on their basis of the original preparations are strongly required. The development of drugs capable of boosting host immune response and affecting target molecules involved in viral pathogenesis seems to be very promising in this respect. Products formulated on the basis of released-active forms of antibodies (RA Abs) meet this needs. The recent years brought persuasive results of RA Abs efficacy obtained in a variety models against a number of viral diseases. For instanse, in a models of influenza, rota–, rhino– and herpes viruses the reduction of pathogen titers, resolution of symptoms along with the higher survival rates were demonstrated. For retroviral infection an inhibition of main viral factors involved in the process of penetration and replication was examined. Apart from that, the drugs demonstrated an excellent safety profile in full-scale toxicity studies.Thus, drugs on the base of RA Abs seems to be a worthy candidates for further development and investigation.
 

Erdogan Esref Hakki

University of Selcuk, Turkey

Title: Triticum monococcum based genes as a source of salt tolerance in Turkish wheat genotypes

Time : 14:20-14:40

Speaker
Biography:

Erdogan Esref Hakki has completed his PhD from Middle East Technical University (Ankara), Department of Biotechnology, in 2000. He is running a Molecular Genetics Laboratory in Konya (TR) within Selcuk University, Faculty of Agriculture. His studies are mainly focussed on abiotic stresses (boron, salinity etc.) of crop plants. 

Abstract:

Worldwide, around 20% of irrigated lands are damaged by salt stress. Hence, either crop production from the non-effected land should be increased or genotypes with stress tolerance should be developed to be grown in stress-affected land. Salt stress largely effects crop yield by making the spikelets sterile, inhibiting the flowering, reducing the grain weight and leading to stunted plant growth. Saline growth environment produces phenotypical symptoms that are linked to physiological and biochemical mechanism of the plant. Wheat, an important cereal crop and chief source of energy is widely consumed in different parts of the world. Apace with the continuously increasing population, it is crucial to enhance its production to meet the current nutritional requirements. However, its production is largely restricted due to salinity stress in arid and semi-arid regions of the world. In such state, developing wheat varieties with greater salt tolerance can be a preferred strategy. Hence, in a combined classical breeding and marker assisted selection program, Triticum monococcum based salt tolerant genes have been transferred in Turkish bread and durum wheat genotypes. In order to identify the other genes involved in the physiological mechanism, Backcross 3 material of the program will be tested for tolerance against high levels of salinity under greenhouse growth conditions. 

Mehmet Hamurcu

University of Selcuk, Turkey

Title: Physiological responses of nitric oxide applications on diverse crops grown under abiotic stress conditions

Time : 14:40-15:00

Speaker
Biography:

Mehmet Hamurcu has completed his PhD from Selcuk University, Konya, Department of Soil Science and Plant Nutrition in 2007. He is running a Physiology
Laboratory in Konya (TR) within Selcuk University, Faculty of Agriculture. His study focused on abiotic stresses (drought, salinity, boron etc.), antioxidant
activities, and reactive oxygen species of plants.

Abstract:

Being a colorless low molecular weight gas molecule, NO plays an important role as a second messenger in biochemical and physiological processes in plants. Researchers determined that exogenous applications of NO molecules are involved in plant defense mechanism in addition to plant growth and development, germination, de-etiolation, chlorosis, and senescence mechanism. NO plays a crucial role in signal transduction and interacts with active oxygen species to inhibit lipid peroxidation in plants under stress conditions. Induction of oxidative properties by reacting with superoxide radicals, especially under drought stress conditions, indicates the potential antioxidant role of NO in plant metabolism. Therefore, it is important to determine the effect of nitric oxide application under stress conditions. Our research group has examined the physiological and biochemical effects of NO donor SNP on different plant species like wheat, rye, barley, maize, beans, soybean and watermelon under different abiotic stress conditions such as drought and salinity. The results of the study showed that NO applications in abiotic stress conditions were effective in suppressing reactive oxygen species and MDA levels, and also had positive effects on protective antioxidant activities that produced stress responses. These effects were more effective in case of wheat, barley, bean and soybean plants.

Saroj Mishra

Indian Institute of Technology Delhi, India

Title: Hyper-production and stability of human serum albumin in Pichia pastoris through a combination of medium design and genetic strategies

Time : 15:00-15:20

Speaker
Biography:

Saroj Mishra completed her PhD from City University of NewYork, USA followed by Post-doctoral research at Institute Pasteur, Paris, France, VTT
Biotechnical Laboratory, Espoo, Finland and University of California, Davis, USA. She has published more than 87 papers in reputed journals and leads a
large group of scientists working in the area of enviromental biotechnology, whole cell biotransformation and recombinant therapeutics.

Abstract:

Human serum albumin (HSA) is an important therapeutic recommended for treatment against trauma, burn injury, hypoproteinemia, hypoalbumenia as well as for maintenance of homeostasis, transportation of hormones and microelements in blood. In this study, we report medium design and genetic strategies that lead to high production of this protein in the culture spernatant of Pichia pastoris. The codon-optimized gene for HSA was cloned downstream of α–factor secretory signal sequence and the mature HSA was secreted in the culture supenatant of P. pastoris under the control of alcohol oxidase 1 promoter. Extracellular protein level of 0.12, 0.40, 1.2 g/L were obtained in the un-optimized medium for 1-copy, 2-copy and 3-copy expression casettes respectively at shake flask level. Factors affecting production were identified which included initial peptone concentration, methanol concentration and temperature, amongst many other (pH, aeration, sorbitol concentration, initial inoculum) investigated parameters. A three level factorial design named central composite design using Plackett Burman response surface methodology was used to optimize the medium which lead to levels of protein up to 0.075, 0.40 and 0.98 g/L total extracellular protein for 1,2 and 3-copy constructs respectively. Under these conditions, HSA produced was stable and free of other contaminating proteins in the culture supernatant. A detailed transcriptome analysis of the recombinant P. pastoris, cultivated on unoptimized and optimized medium lead to identification of several protein coding transcripts which were up-regulated and helped in efficient HSA production and secretion. These were mapped to biochemical activities linked (and not restricted to) to carbon, nitrogen metabolism, gene transcription, protein transport and secretion. Additional genetic strategies applied included modification of signal sequences. Application of optimized medium to these mutants lead to stable production of HSA with reduced proteolytic degradation of the synthesized protein. This illustrated the robustness of the designed medium with a production of over 2 g/L protein at shakeflask level. An understanding of the underlying mechanisms is likely to play significant role in use of Pichia system for production of heterologous proteins.

Christopher Alexis Cedillo-Jimenez

Autonomous University of Queretaro, Mexico

Title: Exogenous application of dsRNA for fruit developmental improvment

Time : 15:20-15:40

Speaker
Biography:

Christopher Alexis Cedillo-Jimenez has completed his Bachelor’s degree in Food Chemistry, Master’s in Science of biosystems from molecular perspective, and a certificated Diploma for Greenhouse Engineering and now he is siding his PhD from Autonomous University of Queretaro. He is the Director of “DETAGS” (enterprise for development of ago-logistic technology) and “Motus collective” (arts, science and technology communication). His thesis work is related to genetic material that contain miRNA sequences related to ripening and developmental targets silencing to improve some qualities of fruits at plant
and postharvest level.

Abstract:

GMO has been proposed as an alternative to improve crops to: increase the yield and production volume, using fewer sources and to resist biotic and abiotic stress. However, the debate and public opinion is still divided, and the market is not ready for GMO related to food production. Interference-RNA knowledge has been used to develop spray-induced gene silencing (SIGS) for pests and diseases control. In this new technology, ectopic application of specific double-string RNA (dsRNA) provide resistance to plants to some pathogens like and verticillium that affect fruit production. Thus, it is an improvement in plant health and consequently in food production yield without using chemical compounds that has negative impact on the environment and even preventing resistant strains induction. However, in our knowledge there is no research nor applications of SIGS technology to elicit and modify plant physiology. Some authors had proven that dsRNA is locally absorbed and then translocated inside the plant, processed like small interference RNA and strikingly remains stable for 168 hours. In this work we discussed the potential use of dsRNA and miR395 interaction to targets potentially related to ethylene biosynthesis as a new engineering genetic without GMOs to delay ripening of climacteric fruits like tomato (fleshy and climacteric model fruit) which would contribute to decrease food wasting and even open new opportunities for postharvest management and agro-logistics.

Valeriya Zabelina

National Agriculture and Food Research Organization, Japan

Title: Standardization of products of biotechnological sericulture by parthenocloning and cryobanking of transgenic clonal silkworms

Time : 16:00-16:20

Speaker
Biography:

Valeriya Zabelina has completed her PhD in the field of Genetics at the age of 26 years from Karazin Kharkiv National University, Ukraine, was involved
in teaching of Developmental biology and worked at the Laboratory of Germ and Stem Cells at the same University, Biology Faculty, Chair of Genetics
and Cytology. Then continued her postdoctoral studies at the Biology Centre of the Czech Academy of Sciences, Czech Republic under several European
projects (Mobitag, Postdoc Bioglobe), containing research stays abroad (Spain, Japan) and presenting her results at international meetings. She was also
involved in teaching of Developmental biology and supervision of a student at the University of South Bohemia, Czech Republic. At the moment she is
conducting her research in Japan under JSPS fellowship with the group working in transgenic silkworm. She has been a member of Ukrainian Society ofGenetisists and Breeders and Japanese Sericultural Society.

Abstract:

Current genetic engineering of the silkworm enables to produce new kinds of silk and to use silkworms as proteosynthetic
bioreactors for obtaining precious proteins. Nowadays a lot of target products derived from the silkworm are already
used in many areas of life from cosmetics through pharmacy, regenerative medicine to cosmonautics. The problem of
product standardization we propose to solve by easy and inexpensive decision – parthenocloning, which enables rapid
fixation of transformed genotypes and conservation of any unique man-made complexes. Parthenocloning is based on
exact copying of maternal genome and a single female initiates a clonal line with the same genetic and morphological
traits, which can be easily maintained without sexual reproduction as exclusively female populations. Such populations
exist already for more than 50 years as genetically stable females whose unfertilized eggs are induced to develop by
heat-shock treatment. This is the best time-tested evidence of genome stability. We constructed new non-diapausing
parthenogenetic strains and developed efficient injection method adapted for the eggs of parthenoclone. We showed
that transgenic silkworms could be obtained in high frequency and propagated as clonal populations. We obtained
successful transgenesis in a parthenoclone and inserted transgenes were faithfully transferred to successive generations.
We showed the possibility of cryopreservation of ovaries of transgenic clonal strains and obtained recovery as individuals
by ovary transplantation into female larvae, which opens door for cryobanking of any specific standardized genotypes
used for obtaining of precious target products.

Yilkal Belay

Makerere University, Uganda

Title: Single dose acute toxicology in preclinical trial: The basic step in drug discovery and development
Speaker
Biography:

Abstract:

The dose, in preclinical trial, refers to the amount of test material administered at once to study subject for
pharmacological evaluation. Acute toxicity study of three different pesticides (Dichlorvos, Chlorpyrifos and
Cypermethrin) with five level of doses each (10, 30, 50, 70 and 90) mg/kg body weight, were evaluated on15 Balb c mice
for a maximum period of 5 days. The main clinical signs and symptoms developed after treatment with five levels of doses
prepared from chloropyrifose was salivation, lacrimation, miosis (pinpoint eyes), trembling and breathing difficulty
with hypo-activity which was significantly manifested within about 30 minutes to 2 hours after treatment depending
on the amount of dose administered orally. Distended stomach, tremor and restlessness, breathing difficulty, salivation
and bulging eyes was the clinical signs of toxicity developed after treatment with the five levels of doses prepared from
Cypermethrin pesticide which was also administered orally. The Balb c Mice treated with five levels of doses prepared
from dichlorvos pesticide developed slow respiration immediately after oral administration. The dose had never limited
the toxic property of test chemicals but the magnitude of adverse effect and length of time at which the undesired
effect manifested in treated Balb c Mice. Even if the higher dose (90 mg/kg) from each test chemical was lethal within
24 hours, the second and third highest doses (70 & 50 mg/kg) which was prepared from Cypermethrin caused lethal
effect in the second day after dosing orally. This implies that the undesired effect of test chemicals was due to its toxic
reaction rate (r) in the biology of treated Balb c Mice. Blood samples from each treated Bulb c Mic were drawn from the
tail and facial vein using micro tubes labeled with numbers and quantitative immunoglobulins test had been conducted
using architect system – Abbot before treatment as reference test and four hours after treatment for comparison. The
toxic reaction rate and toxic severity of each test substances was then calculated using the formula [r =-(± lg plasma
concentration)] in mg/sec and (s = x 100) in %/sec respectively and recorded in different tables. The study revealed
that the value of toxic reaction rate (r) determines the margin of safety whereas the value of toxic severity (s) of test
chemicals predicts the length of time at which lethal effect of test substance might be manifested in treated Balb c Mice.
Tested doses with calculated value of toxic reaction rate (r) less than zero had no lethal effect to treated Balb c Mice. This
means that the administered test chemicals had negligible adverse effect at the organismal level rather than at the cellular
level. A test substance said to be toxic not only when it causes death but also pharmacological mechanism against the
biology of an organism. If the higher dose kills treated organism, the lower dose is most likely to have a higher risk of ill
health in the long run. There is no scientific ground to categorise a single test material as safe dose (ED50) and lethal dose
(LD50). It is most likely to be a waste of time and resources to categorise a single test chemical as effective dose (ED50)
and lethal dose (LD50) at a period of time during the experiment and proceed to the next phase of preclinical trial with
inadequately validated data. The presentation will have more details on single dose acute toxicology in preclinical trial.

  • Oncolytic Biotechnology| Molecular Biotechnology and Genetics| Microbial Biotechnology
Location: Winter garden
Speaker

Chair

Sergey Suchkov

I M Moscow State Medical University, Russia

Session Introduction

Mihai Gidea

University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania

Title: Research on the testing of products with biostimulatory effect based on amino acid with potential in the treatment of rape seed
Speaker
Biography:

Mihai Gidea is form University of Agronomic Sciences and Veterinary Medicine of Bucharest, Romania

Abstract:

Due to its major potential for biofuel production, the rape-cured areas have steadily increased lately, reaching a cultivated area of 38 million ha worldwide at 2016 (FAOSTAT). In these conditions, the problem of increasing the
level of the obtained productions is raised more and more, The paper presents the results of the researches regarding the treatment of rape seeds with products containing amino acids (aa). In this context, there were 4 products based on keratin hydrolyzed wool and chelated (co) of Zn, Mn, Cu, Mg and Mo. For testing, a bifactorial experience was performed where Factor A tested the product with 5 graduations a1 14% aa + 0.5% co, a2 12% aa + 0.3% co, a3 10% aa + 0.4% co, a4 14% aa+0.4% co+1% Caryophyllus aromaticus oil microcapsules, and Factor B seed immersion time in products tested with a1 control, a2 1h, a3 2h, a4 3h. The assays were performed under laboratory conditions. The treatments were film-coated and, after treatment, the seeds were seeded, the TopPaper recommended by ISTA for rapeseed testing.The research found that all the treatments applied had a stimulating effect on the monitored parameters, thus increased the rate and germination rate, and increased the average daily germination rate, the average germination time. From a biometric point of view, there was an increase in the average length of plantlets and roots, as well as the average daily growth rate. The treatments applied did not show phytotoxic effects.

Maryam Ranjpour Aghmiouni

Jamia Hamdard University, India

Title: Elevated expression of cytosolic phospholipase A2 delta is associated with hepatocellular carcinoma progression: Animal study validated with sera of liver cancer patients
Biography:

Maryam Ranjpour Aghmiouni has completed her PhD from Jamia Hamdard University. She has applied for Post-doctoral position at Illinois University
and her application is under process. She has published two manuscripts and three more manuscripts are under re-revision status at high repute
journals.

Abstract:

Liver cancer is the third common cancer to cause maximum death among patients diagnosed with cancers. The search for new biomarker discovery is necessary as none of the identified biomarkers alone are enough sensitive toward hepatocellular carcinoma (HCC). In order to find out novel biomarkers that can diagnose HCC at very early stage, we have developed a rodent model using chemical carcinogens, N-Nitrosodiethylamine (DEN) and 2 aminoacetylfluorine (2AAF). The disease progression was monitored by histological evaluation. Proteomic approaches such as 2D-Electrophoresis, LCMS/MS and Western blot analyses have been used to analyze the differentially expressed proteins in carcinogen-treated animals vis-a-vis controls. The total serum proteins were isolated, solubilized and resolved on 2D-Gel Electrophoresis using broad pH range IPG strips. PD-Quest analysis revealed proteins that are differentially expressed in serum of the carcinogen-treated rats as compared to controls. Some of these proteins have been identified by LCMS/MS. Histological analysis confirmed liver inflammation and disease initiation at one month and tumorigenesis at four months after carcinogen treatment. One of the differentially expressed proteins, namely, cytosolic phospholipase A2 delta was significantly up-regulated at very early stage of cancer development and continued to remain elevated with disease progression up to tumor stage. The increase in its expression has been confirmed by Western blot analysis. Further, the analysis of serum of liver cancer patients also showed elevated expression of this protein that validated our experimental data. The study suggests that elevation in cytosolic phospholipase A2 delta expression is associated with progression of HCC.

Ekei Ikpeme

University of Calabar, Nigeria

Title: Single nucleotide polymorphisms and haplotype analyses in tilapia fish inferred from mtDNA D-loop and Cyt-b regions
Speaker
Biography:

Abstract:

Objective: The research was aimed at analysing single nucleotide polymorphisms and haplotypes on D-loop and Cyt-b regions of the mitochondrial DNA of tilapia fish.
Methods: Fifteen and thirteen tilapia fish were obtained from two populations, South-South (Domita farm) and South West (Odeda farm). DNA extraction from fish tissue was done using Quick-gDNATM mini prep kit
after which PCR amplification was carried out. Sequencing of the two mtDNA regions were done using forward primer 5’- GGATTYTAACCCYTRCCCC- 3’ and reverse 3’-AGTAAGTCAGGACCAAGCC-5’ for D-loop and
5’-GGATTTTAACCCTTACCCC-3’ and 3’AGTAAAGTCAGGACCAAGCC-5’ for Cyt-b region. Statistical analyses were carried out on the aligned sequenced data using MEGA version 6.06, DnaSP 5.1, Codon code aligner 6.06 as well as NETWORK 4.6.1.1.
Results: mtDNA polymorphism was highest in the D-loop of South-South (SS) population with 176 polymorphic sites, while South-West (SW) population had 162 polymorphic sites translating to 176, 162 and 144 SNPs with nonsynonymous substitutions higher than synonymous substitutions. Haplotype diversities (Hd) were 1.00±0.024 and 1.00±0.030 while nucleotide diversities were 0.168±0.086 and 0.161±0.084 for D-loop of SS and SW populations, respectively. For Cyt b region, haplotype and nucleotide diversities were 0.91±0.003 and 0.051±0.016. Positive selection was more on mtDNA D-loop of tilapia sampled from SS than those from the SW as well as Cyt-b region of tilapia fish from SS. 28 haplotypes were identified among the tilapia from SS and SW with no shared haplotypes while 9 haplotypes were identified from the Cyt-b region with haplotypes 4, 5, 6 and 7 shared between species. Median joining network analysis revealed population-based clustering pattern. Demographic expansion was not significant using Tajima’s D and Fu’s F statistics.

Ferhat Djoudi

Abderrahmane Mira University of Bejaia, Algeria

Title: BACTEC MGIT 960 system and classic Lowenstein-Jensen culture in the diagnosis and drug susceptibility of Mycobacterium tuberculosis from pulmonary specimens, at the Pasteur Institute of Algeria
Speaker
Biography:

Ferhat Djoudi has completed his PhD on Epidemiology and molecular characterization of MRSA in 2015 at Abderrahmane Mira University of Bejaia, Algeria. And he started Postdoctoral studies at the same university, on MDR and XDR tuberculosis in Algeria. He is the Head of Microbiology Department and Teacher-Researcher at the same university. He has published many papers in reputed journals.

Abstract:

Tuberculosis is an old infectious disease and the causative agent is Mycobacterium tuberculosis complex. The direct diagnosis stills long and fastidious since bacilloscopy, even if is fast, lacks sensitivity. The culture on Lowenstein- Jensen (L-J), which remains the reference method with a good sensitivity, sometimes takes up to ten weeks to obtain the result. In order to compensate the slow growth of cultures on solid media, new automated methods have been developed, including BACTEC MGIT 960, Versa TREK, MBRedox, BACTEC 460, which allow early diagnosis and more suitable for antibiotic therapy, in addition to their good sensitivity and specificity. The aim of this study is to verify the contribution of BACTEC MGIT 960 in the diagnosis of pulmonary tuberculosis, compared to basciloscopy and classic culture on L-J medium, at the Tuberculosis and Mycobacteria unit in Pasteur Institute of Algeria. Nine hundred and fourteen (914) specimens were collected between January 2016 and April 2017. One hundred and seventy nine (179) cases were reported positive by L-J classical culture and/or BACTEC MGIT 960. Among the 179 cases, 155 were detected by the BACTEC MGIT 960 system, and confirmed by Ziehl control, L-J subculture and MPT64 immuno-chromatographic assay. On classic L-J culture and bacilloscopy, nevertheless, only 123 and 95 specimens respectively were positive. These results confirm the height susceptibility of BACTEC MGIT 960 in improving the diagnosis of tuberculosis in bacilli-poor specimens, compared to classic culture (p=0.037) and direct examination (p=0.014). Furthermore, the contamination rate was higher in L-J culture: 81/914 (8.86%), including 7 bacilloscopy positive specimens, whereas, with BACTEC MGIT 960, only 29/914 (3.17%) specimens were contaminated, with no positive bacilloscopy cases. This result was statistically confirmed (p<0.0001). However, on the 95 bacilloscopy positive specimens, 6 did not give positive cultures neither on BACTEC MGIT 960 nor on L-J. The main advantage of BACTEC MGIT 960 is its ability to shorten the time of mycobacterial growth to an average of 7 days, compared to the solid medium. Nevertheless, the bacilloscopy and culture on L-J remains complementary to this automat, for a reliable diagnosis. Despite the good laboratory practices, there is an incompressible risk of contamination.