Day :
- Viral Infections | Bacterial diseases | Antimicrobial Agents and Antimicrobial Activity | Fungal Diseases | Pharmaceutical Microbiology | Engineering Biotehnology
Location: Waterfront
Chair
Godfred Menezes
Hail University, UAE
Session Introduction
Vasundhara Rangaswamy
Stanford Hospital and Clinics, USA
Title: Comparing diagnostic microbiology in 2 worlds
Time : 11:20-11:45
Biography:
Vasundhara Rangaswamy completed her MD from India and MS Clinical and Molecular Biology from California. She currently works in the clinical microbiology lab of Stanford Hospital and Clinics, California. She has been involved in lab capacity building in India, Cambodia and Ethiopia and also in public health initiatives in rural India for last 20 years. She has also been involved in teaching clinical microbiology and rural health issues to all levels of health care professionals.
Abstract:
Aim: Microbiology is the pillar of infectious disease diagnosis and treatment. Strengthening of services globally is mandatory to achieve health goals laid out by many organizations. However, diagnostic microbiology services offered even for diseases like Malaria, HIV, TB or diarrhoeal diseases that are prevalent in poor countries, differ remarkably between various labs. The presentation attempts to compare an advanced microbiology lab in California, with labs Cambodia, Africa and rural India.
Methodology & Observations: The observations are based on personal experiences of working in or visiting labs in different countries and settings. For any lab, compared to routine tests offered in haematology, biochemistry, serology, and urinalysis, providing reliable basic microbiology services is challenging. In less developed regions, it is a herculean task. Some of the many challenges faced are, lack of awareness among physicians and common folk about the contribution of clinical microbiology towards patient care, absence of or little regulation on QA/QC systems, formidable costs of infrastructure and unreliable supply of water or electricity. One is confronted with many burning questions. Is the disparity in different settings fair? Are the goals same? Whose responsibility is it to step up lab capacity building? Despite these obstacles, significant changes are taking place in some health care centers due to the efforts of one odd passionate microbiologist, pursuance by astute physicians, remarkable work of organizations like LabCAP, DMDP, CDC, etc., by inventors who have simplified diagnostic tools and made them affordable yet reliable and by global pressure to step up containment of diseases.
Conclusion: More of us need to get involved; efforts need to be constant and probably faster to keep pace with the growing population and spread of diseases.
Godfred Menezes
Hail University, UAE
Title: Evaluation of synergistic activity of lactoferrin with antibiotics against drug resistant bacterial pathogens
Time : 11:45-12:10
Biography:
Godfred A Menezes is currently working at RAK College of Medical Sciences (RAKCOMS), RAKMHSU, UAE. He was an Assistant Professor and Scientist at Hail University, Saudi Arabia; In-charge/Scientist of Central Research Laboratory (CRL) and; Assistant Professor of Microbiology at Sree Balaji Medical College & Hospital, Chennai, India for three years. He has also worked as a Scientist in the Department of Medical Microbiology and Infectious Diseases, Netherlands. He has been worked extensively on molecular characterization of antimicrobial resistance in clinical bacterial pathogens. He has been a faculty of several medical institutes and also a Para-Medical Institute.
Abstract:
The multidrug-resistant (MDR) bacterial infections have escalated as one of the world's utmost health issues. The progress of novel antibiotics has declined over the last half century. The aim of this study was to determine the effect of lactoferrin (human, bovine and camel) on minimum inhibitory concentrations (MICs) of important antibiotics in use against drug resistant bacterial pathogens encountered in the region of Hail, Kingdom of Saudi Arabia (KSA). Totally 147 clinical bacterial isolates were successfully isolated. Pathogens included were: Methicillin resistant Staphylococcus aureus (MRSA)-30 isolates; methicillin resistant coagulase negative Staphylococcus-30 isolates; extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae-40 isolates; fluoroquinolone resistant gram negative pathogens-30 isolates; multidrug resistant Pseudomonas species- 05 isolates; carbapenem resistant gram negative pathogens- 05 isolates; AmpC beta-lactamase producing gram negative pathogens- 05 isolates; vancomycin resistant enterococci (VRE)- 02 isolates. The methods employed were MALDI-TOF for identification, MicroScan WalkAway system for identification, susceptibility testing and LF synergism testing. PCR-Sanger sequencing was done (before and after exposure to LF synergism) to study the molecular biology aspects of the study. In our study, the reproducible synergism effect of LF with antibiotics was found to be remarkable. To specify the phenotypic effects of LF in synergism with antibiotics: the isolates producing ESBL had turned non-ESBL; quinolone resistant isolates had turned susceptible; MRSA had turned MSSA (methicillin susceptible) and VRE had turned susceptible. The molecular biological study suggests changes only in the gene expression after the exposure to LF compounds. The results of this study demonstrated similar effect with comparable results for the LF tested from three different sources (human, bovine and camel). The outcome knowledge of the study would certainly help the Ministry of Health (MOH) in planning the LF adjuvant treatment methods for locally faced drug resistant pathogens causing different infections.
Anne Elain
Université de Bretagne-Sud, France
Title: A fluorescence-based method for the assessment of polyhydroxyalkanoates (PHA) production
Time : 12:10-12:40
Biography:
Anne Elain received her PhD degree in Process Engineering from Rennes I University, France, in 1999. She then joined the Biomaterials and Nanotechnologies Lab (now IRDL) of Bretagne Sud University, France. Currently, she leads the Department of Biochemical Engineering. Her general research interest areas are applied microbiology and fermentation technology for the production of high value-added products and process optimizing (yield, sustainability, economic cost, etc.).
Abstract:
Polyhydroxyalkanoates (PHAs) are polyesters acting as energy and nutrition reserves for many prokaryotes. The substrate, cultivation strategy and production strain, will determine the chemical nature and the physical characteristics (i.e., molecular weight, tensile strength, melting temperature, degradability timelines, etc.) of these polyesters. This opens the door for substituting petrol-based thermoplasts, elastomers or latexes, in multiple industrial and biomedical applications. For instance, in the biomedical field, the great adaptability of PHA, in association with their biocompatibility and their bio-absorption capabilities, make them attractive for use as in vivo implants, regeneration devices or drug delivery systems (3, 4). Until now, the major challenge is to reduce the cost of the biosynthesis process to permit the development of a reliable and sustainable PHA production chain. One aspect of major economic importance is to carefully optimize the operating conditions in order to maximize biomass growth and polymer yield. Hence, a rapid and reliable method of screening and monitoring process performances (i.e., cell growth and PHA contents), is needed. Although, GC-MS analysis provides the most accurate results relative to PHA quantification and monomer composition, it involves extraction and derivatization steps which are complex and time-consuming when applied to a large number of samples. In this study, we developed a method that used the lipophilic fluorescent probe Nile Red (1 mg L−1 solution in DMSO) directly into the culture broth to stain the PHA inclusions inside bacterial cells followed by detection of the emitted fluorescence by both microscopic and spectrometric techniques. Epifluorescence microscopy provides a rapid tool to distinguish producing from non-producing bacterial species and the relative fluorescence intensity (FI) determined at the maximum of emission spectra in the wavelength region of 560-710 nm, allows a fast assessment of the cultivation condition and physical process that may enhance PHA production yield. The method was found effective to select bacterial strains efficient for PHA synthesis among a marine collection. Subsequently, the NR assay was used to determine the C0/N0 ratio of the producing media that may improve the polymer yield as well as to follow the time course of fermentation. The coupling of fluorescent dye staining to epifluorescence microscopy is thought to open up new possibilities for high-throughput screening applications and identification of novel PHA producers.
Ka Tik Cheung
Tung Wah College , Hong Kong
Title: Evaluation of domestic dogs and cats oral flora in relationship of oral hygiene and detection of multi resistant bacterial strains
Time : 12:40-13:10
Biography:
Ka Tik Cheung is a Lecturer at Tung Wah College since 2013. He has completed his Doctoral degree in Microbiology and has several years of experience in clinical laboratory. He has also acquired the license to practice which is certified by The Hong Kong Medical Laboratory Board. His research focuses on “Veterinary infectious disease and antibiotics susceptibility on companion animals”. He is also actively involved in zoonotic disease research. He is a member of Hong Kong Veterinary Nursing Association.
Abstract:
Recently, human-animal interaction has occurred more frequently, animal bite injuries have become a serious and high-risk problems as the result. Oral flora can be transferred by close oral contact and through bites. While most of the bites do not require medical progress, only some bites would become an infection. The resultant infection is typically a poly-microbial infection; consist of common environmental flora and the oral flora in animals. As oral hygiene is an important method to reduce the number of bacteria of human oral cavities, but there are only a few articles demonstrated in domestic dogs and cats. To evaluate the effectiveness of oral hygiene in domestic dogs and cats, this study compared the complexity of isolates from the oral cavities and the frequencies of performing oral hygiene. The age of domestic dogs and cats was compared to the number of isolates from the flora as well. Besides, the oral flora was identified and the frequency of occurrence was evaluated, in order to attempt the bacteriology of domestic dogs and cat’s oral cavity and the causative agent of human infection. In this study, gram stain, a series of biochemical tests and Matrix-assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry (MALDI-TOF-MS) were used in identification. Pasteurella species and Streptococcus species were isolated in high frequency. Neisseria species, Pseudomonas species, Enterobacteriaceae family, Corynebacterium diphtheriae, Achromobacter xylosoxidans/denitrificans and Sphingomonas paucimobilis were also identified in the samples as well. Moreover, detection of multiple antibiotic resistant bacteria was also carried out in this study as well. It was used to provide more evidence-based information on antibiotic therapy in dog bite wounds and related infections. In which, multi-resistant organisms and extended-spectrum beta-lactamase (ESBL) positive Enterobacteriaceae were found in the oral cavity of sample dogs. Antibiotic susceptibility patterns for some bacteria were evaluated in this study.
Debabrata Biswas
National University of Singapore, Singapore
Title: Group A Streptococcal CXC chemokine-protease cleaves the human antimicrobial cathelicidin peptide LL-37 resulting in the loss of immunomodulatory functions of the peptide
Time : 14:10-14:35
Biography:
Debabrata Biswas completed his PhD in the field of Cell Biology while working on Arsenic Toxicity in blood. He shifted his interest to the study of pathogen biology and infectious disease during his Post-doctoral study at Hebrew University of Jerusalem. He joined the laboratory of Prof Emanuel Hanski, working on the mechanism of pathophysiology of group A streptococcal soft tissue infection. His present work in Microbiology department of National University of Singapore deals with bacterial virulence factors and various strategies that might be designed against the disease based on the mechanism of actions of these factors. He is currently investigating molecules in the host inflammatory and immune signaling cascade that might act as the potential targets of the bacterial serine protease ScpC, which is a major virulence factor in soft tissue infections.
Abstract:
Statement of the Problem: Severe-soft tissue infections caused by group A streptococci (GAS) are characterized by a rapid dissemination of GAS followed by massive necrosis and tissue destruction. The human antimicrobial peptide, LL-37, is expressed during invasive GAS infections. It is believed that LL-37 antibacterial activity limits GAS spreading, since mice deficient of the LL-37 murine analog, CRAMP, is more sensitive than wild type mice to subcutaneous GAS challenge. LL-37 also directly recruits neutrophils to the site of infection and stimulates interleukin-8 (IL-8) production by keratinocytes. Thus, the immunomodulatory activity of LL-37 is aimed to exacerbate neutrophil response that is crucial for eradication of GAS from soft-tissue. Yet, analysis of debrided human soft-tissue samples revealed the coincidence of LL-37 along with viable GAS.
Theoretical Orientation: The GAS CXC-chemokine protease ScpC plays a central role in virulence through IL-8 cleavage preventing recruitment of neutrophils to the site of infection and reducing the production of neutrophil extracellular trap. Because of its immunomodulatory activities, we hypothesized that LL-37 may also serve as a substrate for ScpC. Functional significance of ScpC-mediated cleavage of LL-37 was studied in vitro and verified in vivo in the mouse model of human GAS soft-tissue infections.
Findings: Here, we demonstrate that immunomodulatory activity of LL-37 is crucial for controlling GAS spreading in soft-tissue. We found that GAS CXC-chemokine protease ScpC degrades and inactivates IL-8 as well as LL-37. ScpC cleaves the first 8 amino acids from the N-terminal of LL-37. This results in loss of its capacity to recruit neutrophil and stimulate IL-8 production by keratinocytes. In summary, the capacity of ScpC to shut down recruitment of neutrophils that is mediated through both IL-8 and LL-37 as well as inactivate LL-37-mediated IL-8 production by keratinocytes reflects a perfect adaptation of GAS to its human host.
Urszula Kosikowska
Medical University of Lublin, Poland
Title: Activity of novel N-substituted-pyrazoles, dichlorobenzoylthiosemicarbazides, 2,4-thiazolidindiones and 1,2,4-triazole-ciprofloxacin hybrids on Haemophilus parainfluenzae and Haemophilus influenzae planktonic and biofilm-forming cells
Time : 14:35-15:00
Biography:
Urszula Kosikowska (PhD) is a Pharmacist, specialist in Medical Microbiology. She is a Lecturer in the Department of Pharmaceutical Microbiology with Laboratory for Microbiological Diagnostics, Medical University of Lublin, Poland. She has published scientific publications in reputed journals, popular articles and conference reports. Her research interests include issues of respiratory microbiota, diagnostics and drug resistance of bacteria and antimicrobial activity of newly synthesized compounds on planktonic and biofilm-forming cells mainly of the genus Haemophilus and other selected members of the family Pasteurellaceae.
Abstract:
Biofilm is a single- or multi-species highly specialized and multi-functional population of microorganisms living on natural or synthetic surfaces with specific biological and physicochemical properties. Biofilm is also an important factor in the pathogenesis of e.g. opportunistic infections. Among the important features of the biofilm cells is their resistance to drugs and other antimicrobial agents and host defense factors. Consequently, this may lead to therapeutic failures. Hemophilia, mainly Haemophilus parainfluenzae and nontypeable Haemophilus influenzae (NTHi), are opportunistic representatives of the respiratory microbiota which are able to live as biofilm and to cause recurrent or chronic and systemic infections (e.g. endocarditis, bronchitis and bacteremia or meningitis). On the basis of minimal inhibitory (MIC) and minimal biofilm inhibitory (MBIC) concentration values the differences in the effect of newly synthesized compounds on planktonic and biofilm-forming cells of hemophilia was observed. Among all the tested derivatives the highest activity against both planktonic and biofilm-forming cells of H. parainfluenzae was estimated for 1,2,4-triazole-ciprofloxacin hybrids and for N-ethyl-3-amino-5-oxo-4-phenyl-2,5-dihydro-1H-pyrazole-1-carbothioamide against both planktonic and biofilm-forming cells of H. parainfluenzae (including ampicillin-resistant strains) and H. influenzae. The basis for the activity of these compounds was the inhibition of enzymes important for DNA synthesis and cells’ metabolism as well as anti-adhesive properties in the early stages of biofilm formation. Other compounds, e.g. thiosemicarbazides and thiazolidinediones did not have a substantial inhibitory effect on both planktonic and biofilm-forming cells. The effect of the tested derivatives partially depends on species or strain of bacteria, compound’s structure and concentration, and time of incubation.
Tseaa Shambe
Benue State University, Nigeria
Title: Postharvest losses of yam tubers in Benue state, Nigeria, West Africa
Time : 15:00-15:25
Biography:
Tseaa Shambe is a Chemist and a Lecturer in Organic Chemistry. He has published papers on chemical composition and structures of some carbohydrates and their degradation by enzymes and acids. He has also worked on the use of bread and composite flour for bread and confectionaries. He is also interested in food toxicology and works very closely with microbiologist.
Abstract:
Statement of the Problem: Africa is the largest producer of yam in the world, with the highest production coming from West Africa. Benue State is the largest producer of yams in Nigeria followed by other States in North Central region. This is because of the rainfall, soil and other climatic conditions that are favorable for yam production in this region. The yams produced form staple food for about 182.2 million people. Postharvest losses of yam tubers (Dioscorea rotundata and D. alata) may occur from a number of causes ranging from improper handling of the tubers to bio-deterioration by microorganisms, insects or rodents. The largest cause of postharvest losses of yam tubers is from microorganisms.
Methodology & Theoretical Orientation: Isolation and identification of microorganisms responsible for the rot of yam tubers was carried out using standard methods of isolation and identification. Optimum temperature of growth was analyzed; pathogenicity test was conducted on the isolates to confirm them as the etiologies of the rot. Plant extract was prepared and incorporated on media plates and used for antimicrobial sensitivity test.
Findings: Four bacteria species (Serratia marcescens, Erwinia carotovora, Klebsiella oxytoca and Pseudomonas aeruginosa) and five fungi species (Aspergillus niger, Rhizopus stolonifer, Botryodiplodia theobromae, Fusarium oxysporum and Penicillium marneffei) were consistently isolated in samples from the various sampling areas. Pathogenicity test revealed the organisms as the cause of the rot. They were inhibited by the plant extracts partially or completely.
Conclusion & Significance: Extensive loss of the harvest can be prevented by blending the various extracts and spraying them on the yams to arrest rot.
Leonardo Madrucciani
Advisor Petroleum LTD, UK
Title: Viral Detection by Nano-amplification Micro Array Technique
Time : 15:25-15:50
Biography:
Abstract:
Statement of the Problem: Microarray as a novel biological technique has been used in gene detection widely. But this method needs improvement, considering expensive probe labeling material and valuable signal scanner. So a visual detection technique was established by applying sandwich hybridization and silver stain. This technique was 100 times as sensitive as LCS (laser confocal scanning) and made it possible to develop a rapid and cheap detection method for Virus. Increasing studies revealed the application of gold nanoparticle as oligonucleotide label was wide.
Methodology & Theoretical Orientation
Materials of Viral detection microarray
Preparation of gold nano-particles described by Grabar (Grabar et al., 1995)
Preparation of detection microarray for Virus
Findings: Cumulative evidences showed gold labeled probe was efficient and could be alternative marker in microarray assay. On the other hand, the sliver stain enhancement, a technique companied with nano-gold labeling oligonucleotide, was used to amplify detection signal of nano-gold. This technique made detection signal visual and its sensitivity of gene detection was approximately 1000-fold high as that of Cy3-based fluorescence.
Muhammad Riaz
University of Agriculture, Pakistan
Title: PCR-RFLP, a diagnostic tool for rapid detection of drug resistant Mycobacterium tuberculosis
Time : 16:10-16:35
Biography:
Muhammad Riaz has PhD degree in Biochemistry from University of Agriculture, Faisalabad, Pakistan and completed his six months research training at University of Glasgow, Scotland, UK. He is working as a Lecturer in the Department of Allied Health Sciences, Sargodha Medical College, University of Sargodha, Pakistan. He has published more than 15 papers in reputed journals. He participated and presented research papers in international conferences as oral and poster presentation.
Abstract:
Tuberculosis (TB) is a highly contagious disease caused by Mycobacterium tuberculosis. Roughly 1/3rd of the world’s population is infected with Mycobacterium tuberculosis (MTB). So, the current study was carried out to determine the drug resistant Mycobacterium tuberculosis magnitude through PCR-RFLP. Initially, the patients were screened for tuberculosis through sputum smear microscopy by Zeihl Neelsen (ZN) staining technique. The sputum positive patients were included in the study after informed consent to the patient. A total of 341 patients were included in the study. PCR-RFLP was used to evaluate the variation in genetic makeup of drug resistant mycobacterium tuberculosis strains. Among the studied population, individuals in older age are affected more and also TB is more common in uneducated and poor people. Resistance against isoniazid, streptomycin, ethambutol and ofloxacin were studied. Overall 91.5% patients were confirmed positive for M. tuberculosis complex infection on PCR analysis. It was found that 17.30% of the cases in the study population have multidrug resistant tuberculosis (MDR-TB). Co-infection of TB with diabetes, HCV and HIV were also observed. Among the drug resistant TB cases 24.04% of the cases were resistant to isonizid (INH), 19.87% resistant to ethambutol (EMB) and 16.99% resistant to streptomycin (STRM). Fluoroquinolone namely ofloxacin resistance along with ethambutol, isoniazid and streptomycin was observed in 6.5% of cases. We concluded PCR-RFLP as a useful molecular technique for rapidly detecting mutations in drug resistant TB patients.
Shimuye Kalayu Yirga
Woldia University, Ethiopia
Title: Preparation and characterization of monoclonal antibodies against citrinin (CTN) for immunoassay VP
Time : 16:35-16:55
Biography:
Shimuye Kalayu Yirga has done his PhD student in School of Life Sciences, Fujian Agriculture and Forestry University in China. Since 2013, he has been working as a Lecturer at Woldia University in Ethiopia. His PhD research focuses on molecular immunology, production of monoclonal antibody and antibody cancer therapy. Recently he found an IgG2a, IgG2b and IgG3 monoclonal antibody against kidney toxin and characterized using different immunoassay. He has done his Bachelor’s degree from Haramaya University, Master’s degree from Bahir Dar University in Ethiopia.
Abstract:
Citrinin (CTN) is a hepato-nephrotoxic mycotoxin produced by fungi genera of Aspergillus, Monauscus and Penicillium. CTN contaminates grains, fruits, juices and vegetables cause various toxic effects to humans and animals. The commonly used analytical methods for CTN detection were TLC), HPLC, LC-MS/MS, (GC-MS). However, these methods have lack of potential for automation, low sensitivity, low specificity, complex equipment’s, incompatible with the real sample, energy and time consuming. Citrinin is a small non-immunogenic toxin with a molecular weight of 250.25 g/mol. Thus, it is necessary to conjugate it with carrier proteins for an immuno-response to generate antibodies. Different methods were applied to conjugate CTN with carrier proteins. Bio coupling method was not satisfactory due to difficulties in removing unconjugated materials through dialysis furthermore; in our study the formaldehyde condensation method was also not effective with the conjugation of CTN-OVA because the result of conjugation was not stable. Finally , in this study CTN was conjugated to bovine serum albumin (BSA) and ovalbumin (OVA), respectively, by amide bonds through active ester method using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). In this study, Female Balb C mice were immunized with CTN-BSA conjugate and Obtained a high anti-serum titer (1:32,000 v:v) when compared to the non-immunized control mouse. Spleen cells of the immunized mice were fused with Sp2/0 myeloma cells to obtain 21H27 hybridoma cell. Ascitic fluid of hybridoma cell was produced in mice abdomen, and purified using caprylic/ammonium sulfate precipitation method. The 21H27 anti-CTN mcAb was the IgG2a antibody subclass, and cross-reactivity results indicated that anti-CTN mcAb is specific to CTN with high affinity (2.0-108 L/mol). Indirect competitive ELISA (IC-ELISA) results showed that the linear range of detection was 0.01–5.96 ng/mL and the IC50 was 0.28 ng/mL with a lower detection limit (LOD) of 0.01 ng/mL. The average recovery was 93.8%-1.6% with a coefficient variation of 1.0%-4.3%. Hence, anti-CTN mcAb secreted by 21H27 hybridoma cell was successfully produced and can be used to detect CTN contaminated feed and foodstuffs.
In this study, 2 mL of sodium citrate and 8.2 pH adjusted with potassium chromate was applied for preparation of good color gold solution. In addition to this, freshly prepared (within one week shelf life) anti-CTN mcAb gold conjugated and optimum concentration of coating antigen were the appropriate parameters for the best achievement of anti-CTN mcAb immuno-chromatographic test strip assay.
Figure 1 Graphic summary of monoclonal antibody production against Citrinin for immunoassay.
- Drug Discovery and Development | Pharmaceutical Biotechnology | Antibiotics
Location: Waterfront
Session Introduction
B Koray Balcioglu
TUBITAK Marmara Research Center, Turkey
Title: Anti-angiogenic peptides
Time : 10:30-10:55
Biography:
B Koray Balcioglu is a senior Molecular Biologist with experiences in Antibody Engineering, Phage Display Technology, Molecular Biology and Recombinant Protein and Antibody Purification. His research projects experience includes the development of antio-angiogenic peptides and recombinant antibodies for cancer therapy, the development of gold binding bi-functional antibodies for biosensor applications. He is interested in new biopharmaceutical drug development, biosimiliars and theranostic agents. He has completed his PhD in Molecular Biology and Genetics Department from Technical University of Istanbul. He has one national patent and one international patent regarding the development of peptides and recombinant antibodies against angiogenesis.
Abstract:
Signal transduction through vascular endothelial growth factor (VEGF) and its receptor play a crucial role in Angiogenesis. Therefore, the development of a biotechnological product with anti-angiogenic properties is widely investigated for fighting cancer. For that purpose, antibodies, recombinant antibodies and peptides are generated or selected from various libraries. Previously, we have discovered anti-angiogenic peptides against VEGF by using a 7-mer phage displayed peptide library. The sequence of the peptides was determined after DNA sequence analysis. The anti-proliferative activity of the selected peptides was monitored in vitro on Human Umbilical Vascular Endothelial Cells (HUVEC). Here, we have synthesized and purified several peptides derived from the original anti-VEGF peptide by changing one by one all the amino acids by Alanine. This process so called “alanine scanning” was done for the determination of critical amino acids responsible in the binding of the peptide to VEGF and the ones necessary for blocking the activity of VEGF. Each of the synthesized peptides was tested in vitro on human umbilical vein endothelial cells for their anti-proliferative effect. With this work, new peptides with higher affinity and anti-angiogenic properties are intended to be developed.
Berrin Erdag
TUBITAK Marmara Research Center, Turkey
Title: Development of biotechnological drug candidates on angiogenesis model in Turkey
Time : 11:15-11:40
Biography:
Berrin Erdag has expertise on Antibody Engineering, Phage Display Technology, Molecular Biology and has successfully completed projects in different field of applications such as Cancer Therapy, Biosensor, in-vitro monitoring. She has a national patent entitled, “Peptide Structures that Bind and Block the Activity of Vascular Endothelial Growth Factor (VEGF)”, where an anti-angiogenic peptide is protected. She has also an international patent entitled, “Recombinant antibody structures binding to and blocking the activity of vascular endothelial growth factor-2 (VEGFR-2/KDR)” US patent No: 9193792, 24/11/2015, China patent No: ZL 2010800690064, 29/06/2016, where two anti-angiogenic recombinant antibodies with a potential anti-cancer properties are expected to be protected. She is the co-leader of a Biosimilar development Landmark project which is the first nationally funded biosimilar project in Turkey. The project consists on the development and production of an anti-cancer biosimilar antibody. Right after the support of the biosimilar project, she has been granted by the Ministry of Development for the development of a Centre of Excellence for Medical Biotechnology. She is still working hard for the development of a biotechnological drug development ecosystem in Turkey.
Abstract:
Statement of the Problem: Even though cancer cells are abnormal, they still require oxygen and nutrients. Angiogenesis, the development of blood vessels, is an essential step in the growth of a tumor. Without vessels, tumors cannot grow to be larger than a small fraction of an inch. Vascular endothelial growth factor (VEGF) and its receptor (VEGFR2/KDR) are major mediators of angiogenesis associated with tumors and other pathological conditons, including age-related macular degeneration and proliferative diabetic retinopathy. It is well known that inhibition of VEGF induced angiogenesis is a valid strategy for the treatment of solid tumors and other disorders in humans. In this context, two peptides and two recombinant antibodies able to inhibit angiogenesis have been developed in MRC.
Methodology: In this study, we identified two single chain variable fragments (scFvs) that directly bind VEGFR-2 and inhibit VEGF-dependent cell proliferation and quantified their receptor-binding affinities. Phage display method was used to construct recombinant single-chain antibodies, which are smaller in molecular size, but still retain the VEGF-blocking property of larger antibodies. Two specific single-chain antibodies (KDR1.3 and KDR2.6 scFvs) recognizing the extracellular immunoglobulin-like domains 1–7 of VEGFR-2 were selected from a V-gene phage display library constructed from mice immunized with the commercially available soluble extracellular domains 1–7 of VEGFR-2.
Findings: KDR1.3 and KDR2.6 scFvs were characterized at the DNA and protein levels by ELISA, DNA sequencing, and surface plasmon resonance (SPR) spectroscopy. Both anti-KDR scFvs bind to sKDR D1–7, block VEGF binding to sKDR D1–7, and show potent inhibition of VEGF-induced cell proliferation in human umbilical vein endothelial cells (HUVECs) by a rat cornea angiogenesis assay (CAA).
Conclusion: Our results demonstrated that KDR1.3 and KDR2.6 antibodies could inhibit angiogenesis via. interaction with the VEGFR-2 extracellular domain. Thus the identified recombinant antibodies may have potential to be used as angiogenesis inhibitors.
Erik Andrade Jorge
National Polytechnic Institute, Mexico
Title: Dioxo/isoindoline derivatives as novel drugs for neurodegenerative disease: Parkinson’s and Alzheimer's disease (in silico, in vitro and in vivo study)
Time : 11:40-12:05
Biography:
Erik Andrade Jorge is a Doctorate student in the Department of Biochemistry at Instituto Politecnico Nacional. He is a Chemist-Pharmaceutical-Biologist and has a Master’s degree in Pharmacology and is currently pursuing Doctorate research in medicine. He has two different research lines, one is cancer cell proliferation and another one is Parkinson’s disease. His work focuses on the rational drug design based on the molecular mechanisms of different pathologies and in the physicochemical properties of the ligands.
Abstract:
Neurodegenerative diseases are a heterogeneous group of disorders that are characterized by the progressive loss of the structure or function of neurons, including the death of neuron cells. Common neurodegenerative diseases include Alzheimer's disease and Parkinson's disease. Isoindoline represents an important family of compounds presents in a wide array bioactive molecules and also it has been seen this kind of compounds has effect in the central nervous system. This is the main reason they have attracted the attention of many researchers, even our work group. Therefore, the aim of the present study was to design and evaluate a series of isoindoline to test their selectivity for the dopamine D2 receptor, and a series of dioxoisoindoline as possible inhibitors of acetylcholinesterase. In the study of the molecular and toxicological properties, dioxoisoindoline and isoindolines showed that possess favorable characteristics as potential drugs. Taking into account the in silico results, the synthesis of the molecule Ia1 (isoindoline) and Da1 (dioxoisoindoline), and their structures were confirmed by IR, 1H and 13C NMR and mass spectroscopy was carried out. The in vivo evaluation for the isoindoline (Ia1) showed that this compound has an effect on the motor activity of male C57BL/6 mice in the MPTP model. While molecule Da1 (dioxoisoindoline) was tested in an intro experiment, results showed that this molecule has the ability to inhibit acetylcholinesterase. These results allow us to try other candidates who might possess the same properties and also perform others studies that can give us more evidence about selectivity.
Bribiesca-Carlos José
National Polytechnic Institute, Mexico
Title: Dioxoisoindolines as potential anticancer agents
Time : 12:05 -12:30
Biography:
Bribiesca-Carlos José is a Medical student at Escuela Superior de Medicina at Instituto Politécnico Nacional. He is working at the Biochemistry Research Laboratory in order to improve his knowledge about the chemistry of life and the synthesis of drugs.
Abstract:
Cancer is a disease that has a multifactorial origin, actually, the scientist in the entire world are working for one effective treatment or a cure, but the idea seems so far because in one-day thousands of new cases are detected and in the moment we only have a small idea of confronting it. The new discoveries to stop cancer have a central element, this is the genetic regulation, and one that has great interest is blocking the function of histone deacetylase which has as their central role withdrawing an acetyl group, which helps temporarily silencing gene expression, as it returns to a state of super curl DNA. Our work comprises selective inhibitors of HDAC8, family member number one of the HDACs, which depends on a zinc molecule to perform its function; this specific protein has been implicated in the development of metastasis in cancer breast, besides participating in the differentiation and proliferation of cells by yet unknown mechanisms. We had been designed two molecules that have structural relation, the results obtained in our tests are positive for inhibition of HDAC8 with a ∆G=-5.290 Kcal/mol, we obtained favorable results in terms of reducing the load of transformed cells at a concentration of 1x10-4 M, in 24 and 48 hours, for one of our two molecules, data were shown by spectroscopy at 540 nm. Our second molecule proved to be nothing effective and opens the gap to a discussion on how to change the structure a little can affect the biological response.
Bribiesca-Carlos José
National Polytechnic Institute, Mexico
Title: Dioxoisoindolines as potential anticancer agents
Biography:
Bribiesca-Carlos José is a Medical student at Escuela Superior de Medicina at Instituto Politécnico Nacional. He is working at the Biochemistry Research Laboratory in order to improve his knowledge about the chemistry of life and the synthesis of drugs.
Abstract:
Cancer is a disease that has a multifactorial origin, actually, the scientist in the entire world are working for one effective treatment or a cure, but the idea seems so far because in one-day thousands of new cases are detected and in the moment we only have a small idea of confronting it. The new discoveries to stop cancer have a central element, this is the genetic regulation, and one that has great interest is blocking the function of histone deacetylase which has as their central role withdrawing an acetyl group, which helps temporarily silencing gene expression, as it returns to a state of super curl DNA. Our work comprises selective inhibitors of HDAC8, family member number one of the HDACs, which depends on a zinc molecule to perform its function; this specific protein has been implicated in the development of metastasis in cancer breast, besides participating in the differentiation and proliferation of cells by yet unknown mechanisms. We had been designed two molecules that have structural relation, the results obtained in our tests are positive for inhibition of HDAC8 with a ∆G=-5.290 Kcal/mol, we obtained favorable results in terms of reducing the load of transformed cells at a concentration of 1x10-4 M, in 24 and 48 hours, for one of our two molecules, data were shown by spectroscopy at 540 nm. Our second molecule proved to be nothing effective and opens the gap to a discussion on how to change the structure a little can affect the biological response.
Urszula Kosikowska
Medical University of Lublin, Poland
Title: Antibiotic resistance of Enterococcus faecalis isolated from the hearts of broiler chickens
Time : 12:30-12:55
Biography:
Urszula Kosikowska (PhD) is a Pharmacist, specialist in Medical Microbiology. She is a Lecturer in the Department of Pharmaceutical Microbiology with Laboratory for Microbiological Diagnostics, Medical University of Lublin, Poland. She has published scientific publications in reputed journals, popular articles and conference reports. Her research interests include issues of respiratory microbiota, diagnostics and drug resistance of bacteria and antimicrobial activity of newly synthesized compounds on planktonic and biofilm-forming cells mainly of the genus Haemophilus and other selected members of the family Pasteurellaceae.
Abstract:
Antibiotic-resistant Enterococcus faecalis present in poultry can pose hazard health serving as the risk of transmitting these strains to humans. The aim of the study was to provide data on antimicrobial resistance in E. faecalis isolated from the hearts of broiler chickens, aged from 3 to 6 weeks. 57 isolates of E. faecalis were investigated. First, the isolated bacteria were identified using MALDI-TOF mass spectrometry (Bruker Daltonics, Germany). Then, the identification was confirmed by genus- and species-specific multiplex PCR. Susceptibility testing was carried out to determine sensitivity to vancomycin and ampicillin (0.125-64 µg/ml), gentamicin (2-1024 µg/ml), kanamycin and streptomycin (4-2048 µg/ml), erythromycin and tetracycline (0.25-128 µg/ml), ciprofloxacin, lincomycin and chloramphenicol (0.5-256 µg/ml) using the minimal inhibitory concentrations (MICs) assessment. Resistance to two or more antibiotic agents was demonstrated. High resistance (>50%) was shown in the isolates including resistance to lincomycin (100%), tetracycline (63.2%), erythromycin (54.4%) and gentamicin (52.6%). Furthermore, high-level aminoglycoside (gentamicin - 1.8%, kanamycin - 1.8%, streptomycin - 5.3%) resistance was noted. Resistance to ciprofloxacin (21.1%) and chloramphenicol (8.8%) was classified as low (<25%). Moreover, a certain percentage of isolates exhibited intermediate sensitivity, particularly to gentamicin (36.8%), erythromycin (29.8%), and ciprofloxacin (19.3%). Vancomycin and ampicillin resistant isolates were not detected.
Oleg Reva
University of Pretoria, South Africa
Title: Mupirocin and batumin – New areas of application of known antibiotics
Time : 13:55-14:20
Biography:
Oleg Reva has obtained his PhD in 1995 from Institute of Microbiology and Virology, National Academy of Science of Ukraine. He has his Post-doctoral fellowship in 2002-2004 in High School of Medicine, Hannover, Germany. He is an Associated Professor in the Centre for Bioinformatics and Computational Biology at the University of Pretoria. He has published more than 90 papers cited in 1700 in other publications. Fields of interest are bioinformatics for biotechnology and medicine, genome sequencing, comparative genomics and molecular docking.
Abstract:
Development of new antibiotics is recognized as a global problem. We have to admit that we have been rather prodigal with the natural resources of antibiotics. The idea that one bioactive compound may exhibit many useful activities by targeting many unrelated molecular targets is getting popularity. This work was focused on two antibiotics, mupirocin and batumin, – synthesized by Pseudomonas, which are active against drug resistant Staphylococcus aureus. Our study was driven by a curiosity: what could be the biological sense for the rhizobacterial Pseudomonas to produce these rather complex and energy expensive compounds, which specifically inhibit growth of bacteria not common for the rhizosphere? Pharmacophore based search, molecular docking and experimental trials demonstrated that these antibiotics may have multiple molecular targets. Pathogenic staphylococci were inhibited by targeting tRNA synthetases and fatty acid biosynthesis. Another possible target was the bacterial MotB flagellar motor protein. It was shown that batumin can immobilize phytopathogenic bacteria leaving them alive but not active. Moreover, possible molecular targets were found among eukaryotic cancer related proteins. Mupirocin and batumin showed an extremely high and specific cytotoxicity against several cancer cell lines including melanoma MEL-1 and Lewis lung carcinoma. By preliminary results it was supposed that these compounds caused a cell cycle arrest at G1/S checkpoint via activation of retinoblastoma protein and down-regulation of the cyclin D1. Our results showed that the same antibiotics, or their target specific derivatives, may find much more proper and broader use in medicine, veterinary and agriculture.
Umadevi Umakanthan
Rajaratnam College for Women, India
Title: Potentiation of Antibiotic with Universal potentiator – A novel and basic invention
Time : 14:20-14:45
Biography:
U Umadevi is an Assistant Professor of Botany. She has published 35 research articles in national and international journals. She is an Editorial Board Member of Virology Research Journal. She has Co-authored a scientific book, “The Multi-science 22nd Century Inventions” and also published book on divinity. She has received awards, fellowships, prizes from government and private organizations. She has chaired international conference organized by World Academy of Science, Engineering and Technology, London on 17th and 18th October, 2016 and got best paper award. She has received Alexander Fleming Award from USA.
Abstract:
Efficacies of newly evolved antibiotics are basically assessed under in vitro by antibiotic sensitivity test (ABST). In this study, recently invented universal potentiator was incorporated with various antibiotic discs with control. These antibiotics were challenged with various virulent bacteria (hospital isolates). By ABST, the zone of inhibition measured. The universal potentiator was found to have potentiated the antibiotics up to 3-166%. This discovery has enormous application to the living beings.
Alexandra Hofer
Vienna University of Technology, Austria
Title: In depth characterization of the metabolic adaption of Penicillium chrysogenum to complex raw material nutrients and specific sugars
Biography:
Alexandra Hofer is recently doing her PhD thesis at the Technical University of Vienna, Institute of Chemical Engineering. She has done her Diploma thesis in Pharmacy, focusing on the development of novel methods in order to describe the glycol-recognitive drug delivery systems of targeted therapy of urothelial diseases. Thus, she has a strong background in biochemical analytics. At TU Vienna, her research focuses on the characterization of different raw material and their impact on process performance, concentrating on filamentous processes.
Abstract:
Statement of the Problem: Processes using Penicillium chrysogenum for the production of antibiotics are established since the 1940’s and are already well described in literature. Generally, these processes are performed in two phases, a batch phase where complex nutrients – mainly corn steep liquor (CSL) are used to enhance spore germination and a fed-batch phase using defined media for product formation. Especially in the batch phase, the mechanism of the positive impact of CSL on growth is still poorly understood and accordingly also the effect of complex media supplements that are transferred in the fed-batch phase. This study focuses on the investigation of CSL and its impact on growth and process performance in both process phases.
Methodology & Theoretical Orientation: Varied methods were developed in order to quantify specific nutrients in CSL with the special focus on amino acids. Batch and Fed-batch cultivations with P. chrysogenum, producing Penicillin V, were performed and analyzed concerning variations in its metabolite pattern. Three main topics were investigated in detail: (1) utilization of complex nutrients by the mold, (2) influence of CSL on physiology and (3) morphology of the fungus.
Findings: Correlations in the beforehand mentioned topics of nutrient utilization; physiology and morphology could be found and lead to higher process robustness. Especially information about limiting amino acids enabled an enhanced process control and led to an improved productivity in the main culture.
Conclusion & Significance: Robustness of fungi fermentations is still a challenge in bioprocess development. Therefore, this contribution focuses on a better process understanding in respect to the influence of complex media ingredients on process performance. Especially the results concerning bioavailability of the CSL and its influence on physiology and morphology of the fungus enable an overall deeper understanding of the fermentation.
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