Day :
- Microorganisms in Pharmaceutical Industry | Microbial Ecology and Next Gen Sequencing | Molecular and Protein based Therapeutics | Regenerative Medicine and Stem Cell technology
Location: UK
Chair
Brice Campo
Medicines for Malaria Venture, Switzerland
Session Introduction
Peter A Kessler
Emory University, USA
Title: Role of regulatory T cells in mother to child transmission of HIV
Time : 11:50-12:20
Biography:
Mother-to-child transmission of HIV-1 occurs in a minority of HIV-infected mother-infant pairs, even without any interventions. The mechanisms that protect the majority of HIV-exposed infants from infection are unclear. T regulatory cells (Treg) have important immunomodulatory functions, but their role in the fetus as well as in mother-to-child transmission of HIV is under-studied.
Abstract:
T cell activation increased rapidly in the first 6 weeks of life, more pronounced on CD8+ T cells; a further increase in activation was observed at the time of weaning from breastfeeding at 6 months of age. In contrast, the frequency of Treg was stable over the first 6 weeks of life (median, 0.5%), slightly decreased between 6 weeks and 6 months (median at 6 months, 0.3%) and then slightly increased between 6 months (time of weaning) and 12 months of age (median, 0.45%). HIV-infected infants had significantly higher frequencies of activated T cells than uninfected infants (P<0.01), as expected. At the time of birth, HIV-exposed uninfected infants had higher levels of Treg, compared to infants infected in utero (Figure, P=0.03). Among infants with negative HIV tests at birth, Treg % tended to be higher in those who were HIV-infected by 6 months of life, compared with those who remained uninfected (median, 1.25% vs. 0.55%).
Julia Nowak Jary
University of Zielona Góra, Poland
Title: Immobilization of therapeutic agents on magnetic iron oxide nanoparticles decreases binding
Biography:
Julia Nowak-Jary graduated with an MSc in Biotechnology (2002) and a PhD in Chemistry (2009) from the Gdansk University of Technology, Faculty of Chemistry, Department of Pharmaceutical Technology and Biochemistry (Poland). In May 2008 she held a research traineeship at the School of Biological Sciences, University of Wales Bangor (UK). Since 2013 she has been working as a lecturer and a researcher at the University of Zielona Gora, Faculty of Biological Sciences, Department of Biotechnology (Poland).
Abstract:
A drug’s affinity for binding blood serum proteins, such as albumin, determines a primary interaction affecting its biological activity. Only the free unbound fraction of a drug can induce a therapeutic effect. A range of effective antimicrobial agents, such as peptides containing N3-(4-methoxyfumaroyl)-L-2,3-diaminopropanoic acid (FMDP), are known to be powerful inhibitors of fungal and bacterial growth in vitro; nevertheless, the use of these compounds in clinics has proven intractable due to their irreversible binding of blood serum proteins, causing complete loss of their biological activity. Another limitation, common to FMDP-agents and a range of other peptide drugs, is low stability in blood serum caused by peptidase cleavage.
Our studies have demonstrated that the described caveats of certain drugs can be significantly reduced by immobilization on the surface of magnetic iron oxide, Fe3O4, nanoparticles (MNPs). Ibuprofen, immobilized on MNPs, was found to exhibit high antibacterial activity even at a very low concentration and in the presence of albumin – in strong contrast to the unattached form of the drug. Furthermore, immobilizing LysNvaFMDP, one of the most potent antifungal agents among the FMDP-peptides, on MNPs decreased its affinity to albumin and other serum proteins compared to its unbound form and resulted in high antimicrobial activity towards bacteria. Finally, our studies proved that the model peptides immobilized on MNPs were more resistant to enzymatic hydrolysis. Together these findings demonstrate the promising utility of MNPs for enhancing therapeutic drug delivery and efficacy.
Seitaro Kamiya
Nagasaki International University, Japan
Title: Study on physicochemical interaction between a variety of saccharide and nanoparticles during freeze-drying and normal drying
Biography:
Seitaro Kamiya has completed his PhD at the age of 27 years from University of Shizuoka and entered employment as assistant professor at a Faculty of Pharmaceutical Sciences Nagasaki International University. He is the senior assistant professor of this university. He has published more than 12 papers as a first author in reputed journals. (Up to 100 words)
Abstract:
There are many reports on the in vivo behavior study of nanoparticles administered to the systemic circulation. However, there are few reports on the preservation of nanoparticles. It is very difficult to maintain the nanoparticle suspension state for a long-term storage because it is thermodynamically unstable. Thus, maintaining a constant state in nanoparticles is an important major issue. A lyophilization method with the addition of saccharides has been utilirized to maintain their particle size of nanoparticles. Despite this method is very predominant, the physicochemical intereaction between nanoparticles and saccharides was not studied till now.
At the present study, trisaccharides, tetrasaccharides, or pentasaccharides were added to the nanoparticle suspensions, followed by rehydration of the samples, which had been either dried normally or freeze-dried. The particle size after rehydration at that time was then measured. In addition, each saccharide was measured using a powder X-ray diffractometer and differential scanning calorimetry (DSC) device. We studied the association between the nanoparticles aggregation and the crystal form of saccharides and their mechanisms by using the obtained results of the data of particle size, powder X-ray pattern, and DSC curves. The particle size of the nanoparticles was maintained when it was freeze-dried, while particle aggregation occurred when normal dried samples were used. In addition, crystallinity of each saccharide was not observed in the in the freeze-dried group but was in the normal dried group.
Kitti Andreidesz
University of Pécs, Hungary
Title: The possible role of cytochrome P450s in non-small cell lung cancer therapy
Biography:
After I completed my BSc in Biochemical Engineering, I graduated as a Medical Biotechnologist at the University of Pécs. Since 2016 I’m a PhD student at the Department of Pharmaceutical Biotechnology at the Faculty of Pharmacy, University of Pécs. My research topic and field of interest is the xenobiotic metabolizing cytochrome P450 enzymes and their role in lung cancer.
Abstract:
Lung cancer is one of the most aggressive form of cancers resulting in more than 1 million deaths yearly. From the two types of lung cancer, non-small cell lung cancer (NSCLC) represents about 88 % of all lung cancer cases. NSCLC can be further defined by recurrent driver mutations. The most common mutations are KRAS and EGFR.
Susceptiblity to lung cancer can be influenced by the metabolic capacity of the lung which strongly relies on cytochrome P450 (CYP) enzymes. Therapeutic drug response is influenced by these enzymes also, as they metabolize many of those drugs.
In this study the expression of drug metabolizing CYPs were measured with qPCR in primary, normal human lung fibroblasts (NHLF) and primary, small airway epithelial cells (SAEC) and compared to two adenocarcinoma cell lines carrying different mutations: KRAS mutant A549 and EGFR mutant PC9.
For their role in drug metabolism CYPs may contribute to drug resistance beside ABC transporters. For this reason adenocarcinoma cell lines were treated with cisplatin, a frequently used chemotherapeutic agent, then the CYP expressions were measured and compared to the non-treated cell lines. To replicate in vivo environment 3D spheroids (A549:NHLF, PC9:NHLF) were set up and CYP expressions were measured.
Beside the obvious differences between 2D and 3D cultures, there were not only differences between the primary cells and adenocarcinoma cell lines, but between the two adenocarcinoma cell lines too.
As conclusion: The possible CYP expression changes in cisplatin treatment might influence the outcome of the therapy especially if combination therapy is being used, and the drug is metabolized by a CYP enzyme. Also the difference in the CYP expression in normal and tumor cells should be taken into consideration during therapy.
Further studies are planned to measure CYP protein level in the previous samples.
RodrÃguez López
University of Vigo, Spain
Title: Effect of a biosurfactant extract obtained from corn in active principles permeation
Biography:
Rodríguez-López, L is a PhD student in University of Vigo. She has completed her Chemistry Degree in 2014, and her Master in Advanced Biotechnology in 2016, both in University of Vigo too. She has coauthored six articles in JCR journals. Moreover, she has collaborated for three months in the Department of Pharmacy in University of Huddersfield (United Kingdom), when she started the study presented under the supervision of Dr. L. Waters.
Abstract:
Nowadays, one of the most important challenges in the pharmaceutical industry is to find additives for formulations which improve the permeation and solubility of drugs. Usually, because of their solubilising and emulsifying properties, it is common to use surfactants in ointments and creams [1]. However, some allergies and adverse effects have been associated with these kinds of substances [2]. In this sense, the study of biosurfactants as an alternative to synthetic surfactants is playing a more important role. Biosurfactants are surface-active compounds, composed of biomolecules, with some advantages in comparison with their chemical counterparts, such as biodegradability or low toxicity [3]. For this reason, the aim of this work was to study the effect of a biosurfactant extract, obtained from a corn-milling industry stream, in drug permeation through polydimethylsiloxane (PDMS). This polymer was chosen as a skin mimic, due to its good correlation with an in vivo situation in a case whereby the penetrant lipophilicity was the prime determinant of compound permeation [4]. During this study, the permeation of five drugs in the presence of biosurfactant at different concentrations were determined, in comparison with a control without the biosurfactant extract. The results obtained have shown the capacity of this biosurfactant extract to mantain or improve drug permeation through PDMS. This is a promising approach in comparison with the results obtaining with chemical surfactants like SDS in the same conditions.
V Veena
REVA University, India
Title: Approaches in identification of suitable natural anticancer and anti-inflammatory molecules
Biography:
Dr. V. Veena is currently working as Assistant Professor and Research Co-ordinator at School of Chemical and Biological Sciences, REVA University, Bangalore, India. Veena obtained her doctoral studies from Pondicherry University, India in 2016. She has 8 years of Research experiences and two year of teaching experience for post graduate and graduate students. Her area of Research includes small molecules of diverse origin for cancer therapy and nanophotosensitisers for cancer therapy. She is recipient of junior and senior research fellow, a prestigious merit-based award given by Council for Scientific and Industrial Research (CSIR) and University Grant Commission (UGC), Government of India. She was also awarded Research Project Assistant in UGC-SAP (special assistant program) by Department of Biotechnology, Pondicherry University, India. Her research contributions was published peer reviewed international journals and she is also serving as a reviewer of some journals.
Abstract:
Statement of the Problem: Several herbal and ayurvedic preparations are currently used to treat the cancer patients. Although, several herbs used in such treatment contains pharmacologically important molecules but they are yet to be identified and their mode of action is not studied in detail. Cancer is defined as a wound that never heals due to its complicated cellular organizations. Thus, the main objective of present investigation was to identify the herbal leads that target the inflammatory tumor environment through modern approaches.
The purpose of this study: India is the major country that is rich in the biodiverse compounds to treat disorders through herbal and ayurvedic approaches. Several natural lead molecules are being reported and continuously being investigated globally. The good examples are being curcurmin and taxol derivatives of natural origin that is effective against cancer and inflammatory disorders. Identification of small molecule drugs from herbs by increasing the selectivity towards tumor is the prime importance of the study. In this context, we have investigated several diverse herbal lead molecules that aimed to increase the selectivity and inflammatory aspects of heterogenic cancer components.
Methodology and Theoretical Orientation: The major photochemical components of herbs used in treatment of various disorders was screened to identify an active components based on preliminary studies. Further, through virtual screening, the compounds were identified against cancer specific targets by computational approaches. The results were validation by in vitro interaction studies and cell-based studies.
Findings: The lead molecules were obtained that can be used for in vivo studies for development of anticancer drugs. The findings also enriched the knowledge regarding mechanistic approach of ayurvedic drugs being used.
Conclusion and Significance: The active herbal constituents that target the multiple pathways which are deregulated in cancer was very useful to for identification of some pharmacologically important lead backbone and other vital components present in the herbs increased the selectively towards cancer could be identified. This study also enriched the knowledge regarding the mechanism of action of some Indian ayurvedic herbal preparations.
Biography:
Peter Henderson obtained his PhD in Biochemistry from the University of Bristol. He has held academic appointments at the Universities of Leicester and Cambridge and is currently Professor of Biochemistry and Molecular Biology at the University of Leeds. He has published over 200 papers, mostly on bioenergetics, membrane transport and antibiotic resistance in bacteria, and many with research collaborators in USA, Canada, Japan, Australia and several European countries. He has been serving as an editorial board member of repute.
Abstract:
Resistance of pathogenic microorganisms to antiseptics and antibiotics is becoming a serious threat to human and animal health. We have characterised a novel membrane protein, called AceI, responsible for the efflux of chlorhexidine from cells of Acinetobacter species, so conferring resistance to a widely-used antiseptic [1]. Genome analyses showed there are similar proteins in many proteobacteria, including pathogens, giving rise to a novel family of drug efflux proteins, designated the Proteobacterial Antimicrobial Compound Efflux, ‘PACE’, family [1].
Genes encoding AceI homologues from 23 species of bacteria were transferred to the pTTQ18 plasmid vector, and transformed into Escherichia coli BL21(DE3) host cells, where the expression of each cloned gene in membrane fractions was identified in Coomassie stained SDS-PAGE gels and in Western blots detecting the His6-terminus of each protein.
Out of all those investigated, seven genes were expressed at levels sufficient for production of proteins at a 30 litre fermentation scale. Each of these was then purified in mg quantities by IMAC. The integrity of the purified proteins was assessed by circular dichroism and by assaying binding to known or putative substrates. Several of the highly expressed AceI homologues conferred resistance to acriflavine, a nucleic acid intercalating biocide. We will discuss evidence showing which common metabolites in bacteria are the natural substrates of PACE proteins.
- Pharmaceutical Nanotechnology | Pharmacokinetic and Pharmacodynamic studies | Drug discovery, development and formulations
Location: UK
Chair
Bożena Szermer Olearnik
Ludwik Hirszfeld Institute of Immunology and Experimental Therapy - PAS, Poland S
Session Introduction
Caroline Jain Beuguel
Laboratoire de Microbiologie des Environnements Extrêmes, France
Title: PHA-producing bacteria from deep sea environments: towards a new generation of therapeutics
Biography:
Abstract:
Poly-hydroxyalkanoates (PHA) are natural polymers, biodegradable and biocompatible, synthesized by many organisms, especially prokaryotes. There are over 150 kinds of these polyesters, accumulated in a wide variety of Bacteria as carbon and energy storage material. PHA granules are deposited intracellularly when microorganisms are cultivated in the presence of an excess of carbon source (carbohydrates, fatty acids etc.) together with a nitrogenous nutrient deficiency.
Due to their biodegradability and biocompatibility, PHA can be used as biomaterials in medical or pharmaceutical fields, and numerous therapeutic micro/nanovectors have already been developed over the past two decades.
The present study highlights short and medium-chain-length-PHA (PHAscl-mcl) production capacities of aerobic and mesophilic deep-sea Bacteria, isolated duringIFREMER cruises from hydrothermal vents in Atlantic and Pacific oceans. According to standardized and optimized fermentation protocols, polymers with original and controlled chemical composition such as poly-3-hydroxybutyrate-co-4-hydroxubutyrate (P3HB4HB) were produced by 7 different bacterial genera, including alpha and gamma proteobacteria.
Next, PHA have been modified by ‘green processes’ to obtain amphiphilic copolymers suitable for biomedical use, before grafting onto metal mesoporous nanoparticles. PHA-metal hybrid complex consists of a rigid iron cage, containing drugs or nucleic acids, coated with a protective organic ligand, made of biobased PHA. This new kind of structure allows better cohesion of the vectors in blood flow, internalization in target pathologic cells and improves stealth against the immune system. Finally, these new PHA-metal complexes, responding to drug delivery concept, are intended to increase the therapeutic benefit while minimizing side effects on the human body.
Muhammad Riaz
University of Sargodha, Pakistan
Title: Biological profiling and hepatorehabilitative potential of Mucuna pruriens in male albino rats
Biography:
Dr. Muhammad Riaz has completed his PhD from University of Agriculture, Faisalabad, Pakistan and part of his PhD research work was conducted at University of Glasgow, Scotland, UK. He is working as Lecturer at University of Sargodha, Pakistan. He has published more than 20 reserch papers in reputed journals and presented his research in various international conferences in United Kingdum, Japan and Pakistan. He has been serving as reviewer of reputed journals.
Abstract:
The present research was conducted to study the biological profile and hepatorehabilitative potential of Mucuna pruriens (MP) seeds using male albino rats as animal model. For the study, methanolic extract of Mucuna pruriens seeds was prepared and antioxidant potential was determined. The seeds extract was screened for antimicrobial activity using well diffusion method. Results showed significant antioxidant potential and antimicrobial activity of extract. For hepatorehabilitative potential, the animals were divided into six groups intoxicated with Carbon Tetrachloride (CCl4) except the normal controls. The test group animals were treated with different doses of methanolic seeds extract. Significant (p<0.05) improvement in liver enzymes like ALT and AST activity was observed in dose dependent manner of seeds extract, while no significant improvement in ALP activity was observed as compared to control group animals. Significant (p<0.05) variation in serum total protein, albumin and globulin concentration was observed. Histopathology of liver tissue was also performed. Serious damage in liver cells (hepatocytes) structures in the CCl4 intoxicated group of animals was observed, while an improvement in the histoarchitecture of liver was observed in the positive control group animals treated with Silymarin and in the extract treated group animals. The study concluded that the Mucuna seeds possess antioxidant and antimicrobial activity with therapeutic potential and may be used for the preparation of hepatoprotective medicines
C Y Tayimetha
Catholic university of Central Africa, Cameroon
Title: Yaoundé from 2009 to 2014) and determination of reference laboratory (Centre Pasteur, Cameroun) role in the surveillance of bacterial resistance to antibiotics
Biography:
Abstract:
The resistance of Neisseria gonorrhoeae to antibiotics as recommended by WHO, poses a real public health problem. Thus, a study of the sensitivity profiles and the determination of the role of the reference laboratory in the surveillance of this resistance were carried out in Yaoundé. The aims of this study were to contribute to the therapeutic management of infected patients with appropriate antibiotics; monitor the resistance of Neisseria gonorrhoeae to antibiotics; limit its emergence in order to preserve the recommended antibiotics. Objectives of the study were to study the susceptibility profiles of Neisseria gonorrhoeae to antibiotics and to determine the role of the laboratory in monitoring this resistance. This retrospective and prospective study, was carried out at Centre Pasteur of Cameroon from 1st January 2009 to 30 September 2014. It consisted of isolation of Neisseria gonorrhoeae strains from the human genital specimens, identification and determination of their resistance phenotypes to antibiotics by the diffusion method in agar medium. This highlighted the role of the reference laboratory in resistance monitoring. A total of 193 strains of gonococci were isolated and identified. The most infected age classes were [20-29] and [30-39]. Men were more infected than women (sex ratio 2.01). Several phenotypes of resistance have been described. High resistance to penicillin G (93.3%), tetracycline (58.5%) and nalidixic acid (17.6%) were observed. Ceftriaxone, azithromycin, spectinomycin, and chloramphenicol were effective at resistance rates of 1.0%, 2.6%, 3.1% and 7.2%, respectively. The overall percentage of strains producing penicillinase is 81.1%. Only Ceftriaxone is still effective among the two WHO recommended molecules in Cameroon. The other (ciprofloxacin) should be monitored. As resistances to nalidixic acid (quinolone marker) have been observed since 2010 and continue to grow up exponentially (25% in 2013 and 50% in 2014). Monitoring Neisseria gonorrhoeae resistances to antibiotics is one of the best strategies to prevent resistances in order to preserve the recommended molecules. Centre Pasteur of Cameroon is the reference laboratory chosen by the WHO for monitoring the resistance of Neiseeria gonorrhoeae to antibiotics in Central Africa.
Hiba Zalloum
The University of Jordan, Jordan
Title: Comparative anti-proliferative effects of potential HER2 inhibitors on a panel of breast cancer cell lines
Biography:
Abstract:
The human epidermal growth factor receptor 2 (HER2) is a member of the erbB class of tyrosine kinase receptors that have been clinically validated as targets for cancer therapy. The gene of this protein (HER2/neu) is found to be amplified in 30% of breast cancers and a variety of cancers. Breast tumors are subtype specific, i.e. breast cancer subtypes have different expression patterns for ErbB receptors. In our initial work we used QSAR equations and their associated pharmacophore models to screen the national cancer institute (NCI) list of compounds and Drug Bank database to search for new promising HER2 structurally diverse inhibitory leads. Inhibitory activities of the resulted compounds were tested against HER2-overexpressing SKOV3 Ovarian cancer cell line and promising IC50 values were scored. In this study we have explored these active hits on different normal and breast cancer cell lines that represent different breast cancer subtypes with distinguished expression level of HER2 and HER1.
Fang Xiao Mei
Chinese Academy of Medical Sciences & Peking Union Medical College, China
Title: Endophytes diversity of medicinal plants and the discovery of their HIV activity
Biography:
Fang Xiaomei has completed her PhD in 2013 from Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College. Her major is Microbial and Biochemical Pharmacy and she has been supported by one National Natural Science Foundation of China (NSFCs No: 31400045).
Abstract:
In recent years, with the development of pharmaceutical research, many studies demonstrated that some medicinal plants showed antiviral activity. Therefore,it will be promising to find new secondary metabolites with anti-HIV activity from endophytes, which may produce the same or similar bioactive compounds as medicinal plants.
In this study, 144 medicinal plants, including 74 plants with antiviral or heat clearing and detoxifying effects and 70 plants with other efficacy, were sampled from different regions, including Yunnan, Guizhou, Shanxi, and Jiangxi province. After surface sterilization of entire samples, endophytic bacteria was isolated from different tissues using 7 isolation media. A total of 2168 strains of endophytic bacteria were obtained in this study. Colonization rate (CR%) were then calculated. The results showed CR% differed in regions, seasons, tissue parts of plant, and isolation media, suggesting Yunnan, spring, root part of plants were the best factors to isolate endophytes, respectively.
After preliminary screening according to phenotypic characteristics, 301 strains were selected for 16S rRNA gene sequence analysis. The 301 strains belonged to 3 Phylum, i.e., Actinobacteria, Proteobacteria, and Firmicutes, including 66 genera, 10 orders and 36 families. Endophytic bacteria of medicinal plants showed relatively high diversity that 301 strains belonged to 117 operational taxonomic units (OTUs) by using 97% similarity as a cutoff. The richness, diversity and evenness were then calculated and showed that the endophytic communities differed among regions, seasons, plant tissues or isolated media. Among them, Yunnan, spring, root part of plants, and M5 medium were the best factors to isolate diverse strains, respectively.
A total of 1681 endophytic bacteria, i.e., 879 strains isolated from antiviral or clearing heat and detoxicating medicinal plants and 802 strains from other effective plants, were selected for anti-HIV screening. 3362 fermentative extracts were obtained from these 1681 strains using two different media. These extracts were then screened using a high throughput screening assay for anti-HIV agents. 324 samples were found positive, of which 110 extracts showed high activities against HIV (inhibition ratio ≥ 95%). 50 positive samples were selected based on lower cytotoxicity to SubT1. As a result, 76% of them (38 strains) were obtained from medicinal plants with functions of antiviral or Heat-Clearing and detoxification.
The possible mechanism of 13 extracts were then investigated with stable high anti-HIV-1 activity using a method of rapidly identifying targets for anti-HIV-1 compounds, and one of the extracts exhibits stable anti-HIV-1 activity and has similar mechanism with MK-0518, showing great potential for further development. The data demonstrated that endophytic bacteria from antiviral or clearing heat and detoxicating medicinal plants have a better capacity in producing anti-viral secondary metabolites.