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EPSA Newsletter

Bringing pharmacy knowledge and students together.

Volume 25 | Edition 3 | April 2018 | @EPSA_Online

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Table of contents

Dear all,

4 Potencial drug-drug interactions: Focus on selective serotonin reuptake inhibitors

We made it to the third edition of European Students’ Science Publication (ESSP). It started with 7 abstracts in the first and continued with 5 abstracts in the second edition and now, we are closing the ESSP chapter of mandate 2017/2018 with the third one, consisting of 9 abstracts!

6 Benzoziazepines as etiological agents in acute poisonings treated at the Emergency Department Clinical Center of Vojvodina 8 In vitro - in silico approach in the formulation and biopharmaceutical characterization of piroxicam buccal patches 10 Intervention to improve the appropriateness of therapy in frail elderly hospitalised patients: results of the pilot study in ULSS 16 of Padua 12 The role of a pharmacist in pharmacovigilance system 14 The identification of the genetic cause of precocious puberty 16 Comparison between the nitrogen oxides inhaled when using regular cigarettes versus IQOS cigarettes (HEETS) 18 Fabrication of a biodegradable nanofiber loaded with lidocaine for hernia repair surgery 20 Microfluidics for polymer encapsulation of drug-loaded mesoporous silica nanoparticles (MSN)

It has been a rollercoaster of open calls, promoting, reading, writing and sending emails but we made it - the result is in front of you. I welcome you to take your time and read about research that is being performed by students from all around Europe, about challenges they have faced and what have they gained by publishing their abstract in this publication. Now it is time for THANK YOUs. To the European Federation of Pharmaceutical Science (EUFESP) for reviewing the abstracts, providing suggestions to get them on a higher professional level and for all the hard work they put into every edition of ESSP. To Cláudia Ferreira, Vice President of Public Relations for all her efforts and time she puts into promotion and design of this publication. To the Educational Department, especially to Eva Shannon, Vice President of Education for suggestions, comments and mostly for the support and motivation. To other Team members who made this edition possible. And last, but not least, to students, who submitted their abstracts and collaborated through the whole process of creating the publication. Enjoy the reading. You will learn something new, get inspired, gain knowledge and, who knows, maybe you will be reading your own abstract in the next edition. Yours in EPSA, Anja Šribar, Science Coordinator 2017/2018


European Pharmaceutical Students’ Association

Potencial drug-drug interactions: Focus on selective serotonin reuptake inhibitors Author: Tamara Kovačević ( Mentor: Božana Nikolić, PhD Institution: Faculty of Medicine, University of Novi Sad

INTRODUCTION: According to the World Health Organization (WHO), depression is the leading cause of disability worldwide, as measured by “years lived with disability” (YLDs). It is a very common disorder in primary care and a costly public health problem. Many patients with depression suffer from other chronic disorders, which means that they are exposed to polypharmacy. There are also patients who do not respond to the initial antidepressant therapy, in which case, combination of antidepressants is indicated. Thus, it is important to focus on potential drug-drug interactions and their outcomes which can include negative effects, but also can be beneficial for the therapy of depression. Recent studies indicate that selective serotonin reuptake inhibitors (SSRIs) are now the initial choice of antidepressant and are the most commonly prescribed group of antidepressant drugs in many European countries. Their therapeutic effect is very similar, but their different pharmacokinetic profiles imply that different medicines in this group show various interaction potential. AIM: The aim of the present study was to identify potential drug-SSRI interactions, to classify them and to define recommendations for their management. METHODS: A cross-sectional database study was conducted. The analysis randomly included 185 outpatients (prescribing ≥2 drugs where at least one of them was SSRI) who visited the Health Center Novi Sad over a 1-month period. All interacting drug combinations were identified according to the Drug Interaction Facts (DIFs) compendium. Accordingly to the compendium, the interactions were classified into categories: severity, onset, documentation, pharmacological mechanisms, potential clinical outcomes and management advice.

clinically significant drug-SSRI interactions, while 32.54% of patients were exposed to potentially augmentative interactions. The most common mechanisms of interactions were pharmacokinetic mediated by CYP2D6, CYP3A4, CYP2C isoenzymes. The most frequent potential clinical outcomes were bradycardia (21.3%), serotonergic syndrome (19.53%), and excessive sedation and ataxia (16.57%). Risk for adverse reactions could be managed by close clinical monitoring of patients or by ordering another antidepressant.

CONCLUSION: Among outpatients, there was a common potential for nonaugmentative and augmentative drug-SSRI interactions. Information based on the result of the present study could be integrated in the existing computerised physician order RESULTS: The mean age of outpatients entry system in the Health Center as a form of was 45,8 (SD±9,8), the majority of the clinical support. population were female patients (69,73%), the mean number of prescribed drugs was 4.35 (SD±2.043), and the mean number of morbidities was 2.5 (SD±1.311). In the study, 44.3% of patients were exposed to potentially | @EPSA_Online

What was the biggest challenge whilst carrying out the research and how did you overcome it? The biggest challenge was analysing all the prescriptions and interpreting all the results. It was not easy to discuss such a high amount of information, and there were so many different directions in which the discussion of Please tell us a little bit more about the results might have gone. Luckily, with the help of my mentor, I managed to stay concise yourself. My name is Tamara and I am a 5th year and clear without missing anything out. pharmacy student at the Medical Faculty, University of Novi Sad. During the past 4 years Anyway, I’m proud that the brainstorming I of my studies, I used my curiosity as much performed gave me ideas for further research. as I could in order to explore all the various Also, in parallel with this, I was working on career options that pharmacy offers and to another research project, so it was quite challenging to manage my time and stay see where I fit in best. focused on both projects.

Questions & answers

I found that what interests me the most are pharmacovigilance, pharmacoeconomics, clinical pharmacy and quality control. I often participate in events organised by Pharmaceutical Students’ Association of Novi Sad-PSANS. Fun fact about me is that, besides pharmacy, my two biggest loves are ballroom dancing and shooting!

In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? I think that the ESSP is an excellent way to showcase your work to your colleagues all over Europe, to maybe give them new ideas and inspire them. Also, you can learn Tell us a bit more about your research a lot of new things by reading other students’ publications, which is great! and its significance. Selective serotonin reuptake inhibitors (SSRIs) are one of the most common prescribed drug groups in therapy of depression, especially in Serbia. There is, however, a lack of evidence about the prevalence of interactions of these medications with other drugs, especially in primary care. The idea was to identify all the potential SSRI-drug interactions in order to give recommendations for therapy improvement. Also, the study was conducted using the prescriptions from the electronic database of the Health Center Novi Sad, which is one of the biggest centers of primary care in this area with about 10 000 patients per day, which means that the results of this study are highly comparable.

My advice for students undertaking research in the future is to, first of all, believe in themselves, to be persistent, because we all know how tough it can get sometimes and last, but not the least, to enjoy and have fun while working on their research!


European Pharmaceutical Students’ Association

Benzodiazepines as etiological agents in acute poisonings treated at the Emergency Department Clinical Center of Vojvodina Author: Sanja Aleksić ( Scientific Coordinator: Vesna Mijatović PhD Ass. Prof. Institution: Emergency Department of the Clinical Center of Vojvodina, Department of Pharmacology, Toxicology and Clinical Pharmacology, Faculty of Medicine, University of Novi Sad

Introduction: Acute poisonings develop as a result of excessive ingestion of particular substances or their combinations in a short period of time. According to the data of the World Health Organization, besides malignant and cardiovascular diseases, acute poisonings are the most frequent causes of death. The most commonly used drugs responsible for acute poisonings belong to the group of psychotropic medications and according to the incidence, benzodiazepines have the highest toxicological significance. Aims: The aim of this paper is to present and describe all characteristics of the patients admitted to the Emergency Department of Clinical Center of Vojvodina from the 1st of July until December 31st 2016, under the diagnosis of acute drug poisonings and in whose blood samples benzodiazepines were detected. Materials and Methods: We conducted one observational study in which the data were obtained from a retrospective review of the medical charts, medical reports and toxicology analyses of blood samples of admitted patients. The extracted data were organised into two groups. The first group of data was representing information about the patients who only used benzodiazepines (BZD) while the second group was representing information about the patients who used a combination of benzodiazepines and other xenobiotics (BZD+X). Results: From the total number of admitted patients (614), 23.45% were treated under diagnosis of acute drug poisoning. Psychotropic drugs were the most commonly found causative agents, corresponding to 81% of all analysed drug overdoses. 84.43%

of admitted patients under the diagnosis of acute drug poisonings with psychotropic agents, abused benzodiazepines. More than one active substance was involved in 85.44% of the cases. The clinical picture of acute poisonings with psychotropic substances was developed in 56.67% of patients from BZD and 71.59% from BZD+X group, while consciousness was preserved in the largest number of patients including patients from both groups (63.33% from BZD and 64.77% from BZD+X). 71% of all analysed patients did not use benzodiazepines as their prescribed therapy. Conclusion: At the observed period, in the Emergency Department of Clinical Center of Vojvodina, medications were one of the most common causes of acute poisonings. The highest percentage of admitted patients abused psychotropic drugs, and the benzodiazepines had the most toxicological significance. Because most of the observed patients didn’t use benzodiazepines as prescribed therapy, the introduction of rigorous control of using and prescribing them could lead to the reduction in the number of acute poisonings in our country. | @EPSA_Online

elevate the quality of life of our patients.

Questions & answers Please tell us a little bit more about yourself. My name is Sanja Aleksić and I am a 5th year student of the Integrated Academic Studies of Pharmacy at the University of Novi Sad. Since 2016 I have been a part of a research team at the Faculty of Medicine and so far, I have conducted three different studies in the field of Clinical Pharmacy and Pharmacology, Pharmaceutical Technology and Molecular Biochemistry. I would like to become a PhD student next year and continue my professional development as a researcher. I like to travel and explore other cultures.

What was the biggest challenge whilst carrying out the research and how did you overcome it? The biggest challenges in carrying out this research were the availability of the data and processing them. In order to get access to the data of admitted patients who undertook benzodiazepines, I had to pass rigorous controls and get the permission of the Ethics Board of the Clinical Center of Vojvodina. The data of my interest was available only via a single computer in the Clinical Center, and I did not have permission to export it, so I had to write all the data in my notebook by hand, and then retype it on a computer. Also, toxicology analyses of patients were available only in paper form, so it required a lot of time and effort to combine all this information. With great patience and hard work I overcame all the challenges and published the research.

I orally presented this paper at the International Medical Congress for young researchers in In your opinion, what is the benefit Saint Petersburg in Russia and got an award of joining ESSP and what advice do you have for students undertaking for the scientific work and presentation! research in the future? ESSP is a great opportunity for all pharmacy Tell us a bit more about your research students to publish their research for the first time and develop necessary skills for future and its significance. Benzodiazepines are well-known drugs scientific publications. Also, I strongly believe prescribed in almost all branches of medicine that the benefit of writing for ESSP is the and because of that, they are widely available possibility to upgrade ourselves by connecting to patients. A large number of patients are with other students and young scientist tempted to abuse these drugs, which presents from the different Universities and countries, a serious clinical and pharmacoeconomic exchanging knowledge and inspiring each problem. The data that 71% of admitted other. patients who abused benzodiazepines, even during the last 6 months, did not use benzodiazepines as a prescribed therapy is really discouraging and demands an urgent call for attention. This implies that we, as future pharmacists, should continue to monitor acute poisonings, and stay in touch with the newest information from this scope in order to reduce the acute poisonings and

My advice to the students who are engaged in the research is to never stop questioning, because somewhere, something incredible is waiting to be discovered!


European Pharmaceutical Students’ Association

In vitro - in silico approach in the formulation and biopharmaceutical characterization of piroxicam buccal patches Students: Nikola Jakovljević (, Anja Vlahović Mentors: assoc. prof. Sandra Cvijić, dr. Marko Krstić Institution: University of Belgrade, Faculty of Pharmacy, Department of Pharmaceutical Technology and Cosmetology, Vojvode Stepe 450, 11221 Belgrade, Serbia

Introduction: Buccal patches are novel oral dosage forms with certain advantages, such as improved patient compliance, controlled drug release and absorption through oral cavity mucosa. Objective: The objective of this study was to assess the effect of formulation factors on biopharmaceutical properties of piroxicamcontaining buccal patches using in vitro - in silico tools. Material and methods: Buccal patches preparation Mucoadhesive buccal patches were prepared with piroxicam. Hydroxypropyl methylcellulose (HPMC) was used as a polymer, whereas the amount of HPMC, presence/absence of plasticiser (PEG 400) and amount of the drug in relation to polymer were varied. The method of preparation was solvent-casting. In vitro dissolution studies In vitro dissolution testing was carried out under biorelevant conditions. Each patch was set at the bottom of a glass with 50ml of artificial saliva and the glass was fixed on an orbital shaker which at the speed of 50 rpm simulated the oral cavity movements. At predetermined time intervals, 5ml of dissolution sample was withdrawn and replaced with equal volume of fresh medium. The samples were analysed by spectrophotometer at 355nm. In silico simulation of drug absorption GastroPlusTM software was used to simulate drug absorption following oral administration. Simulations were performed using ACAT model and OCCATT model for the oral cavity. Evaluation of mechanical characteristics The mechanical characteristics were evaluated

by measuring maximum force applied on the films, maximum stress, and calculating break stroke strain percentage. Results: The results indicated that changes in composition of the patches (variation of HPMC, presence/absence of PEG 400 and amount of the drug), and small volume of saliva are the limiting factors affecting the release rate of the poorly soluble piroxicam. The In vitro study showed that an increase in the amount of polymer increased drug load, and thickness of the film decreased drug release rate. In silico simulations revealed that piroxicam would be mainly absorbed from the oral cavity (89.2 %). Changes in drug release rate additionally modulate the rate of drug absorption from buccal patches.The range of break force for thinner films was 2.5922.49 N and for thicker films 10.14-63.03 N, which indicates that the amount of polymer significantly increased firmness. Presence of PEG 400 in some films increased the break stroke strain. Conclusion: In vitro - in silico assessment of the model formulations indicated that biopharmaceutical properties of a drug and formulation composition are the key factors governing bioperformance of buccal patches. A suitable buccal patch formulation strategy might provide controlled drug release. | @EPSA_Online

yet. Hence, the analysis of published research was very time consuming, as well as finding the right methods and adjusting them in accordance with our available materials.

Questions & answers Please, tell us a little bit more about yourself. My name is Nikola Jakovljević, and at the moment I am a 5th year student at the Faculty of Pharmacy in Belgrade. The thing I am most passionate about is gaining knowledge and because of that, my research work began when I started learning about pharmaceutical technology and industry. I am a hard working person and I always try to do different things! Therefore, I am part of several students organisations and I have been doing a lot of volunteering. Tell us a bit more about your research and its significance. Since 2016 I have been dealing with finding the right formulation of buccal patches (oral films) which could fulfil desirable expectations related to drug release and compliance. After the formulation analysis, examinations on its in silico and in vitro characteristics were performed. At the Conference of Biomedical Students of Serbia, my oral presentation for this research was awarded with the first place award! I truly believe that buccal films are the future in solid dosage forms, especially for certain population groups. What was the biggest challenge whilst carrying out the research and how did you overcome that? Because this is a novel solid dosage form, there are no specific demands for buccal patches approved by any Pharmacopoeia regarding needed examinations and criteria,

In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? I find this an excellent opportunity for all young people who see themselves in future scientific work, to do this as a first step and make their recent work visible to others. This will definitely help them obtain new contacts and prepare themselves for future jobs. I warmly advise everyone in scientific work to choose a field that they are good at and to make their ideas happen. Even though it might seem hard at the beginning, hard work always pays off!


European Pharmaceutical Students’ Association

Intervention to improve the appropriateness of therapy in frail elderly hospitalised patients: results of the pilot study in ULSS 16 of Padua Students: Martina Tammaro, Supervisors: prof.ssa Maria Cecilia Giron, dott. Umberto Gallo, dott. Daniel Dumitru Tinjala Institution: University of Padua Faculty of Pharmacy, Sant’Antonio hospital in Padua

Introduction: In international literature, the appropriateness of therapy is related to adverse drug reactions, clinical important hospitalisations and welfare costs that are not supportable. Potentially inappropriate prescriptions are very common in geriatric patients and the improvement of those is the future challenge for the Health systems. Aim: Firstly the aim of the project SAFE is the “reconciliation” of drug therapy, and the analysis of both prevalence and type of potentially inappropriate prescription in Sant’Antonio hospital in Geriatric ward. Secondly, research aims at the evaluation of the importance of the pharmacist in ward. The pharmacist with the help of a validated “self-built software” analyses the therapy and discusses it with involved clinicians.

Index and 67% of them were in polypharmacy (≥5 drugs). 1186 patients presented prescription potentially inappropriate, and after the reconciliation there was a statistically significant reduction in the MAI/patient score [4 (Inter Quartile Range=2-6) vs. 2 (IQR=05); p <0.001]. In particular 54% of patients improved appropriateness of therapy, STOPP criteria improved in 49% and drug-drug interactions improved in 56% of patients. All the risk associated with PPI significantly Methods: Anagraphic and therapeutic data decreased during the project. were collected for each patient included in the study, those data were entered in the software, Conclusion: Frail elderly are prone which detects prescriptions potentially to clinically important ADRs, which are inappropriate (PPI) identified by STOPP associated to polypharmacy and inappropriate criteria and/or all clinically relevant drug-drug drug prescriptions. interactions (DDIs), identified by Micromedex. It is possible to improve the appropriateness In the presence of PPI the software generates of prescriptions in term of reducing MAI score a report, which the pharmacist discussed with per patient with the presence of pharmacist in the physicians. ward and using the validated software. MAI (Medication Appropriateness Index) was calculated by the software for each patient before and after the intervention. The difference between the two values indicates whether the therapy was appropriate or not. The original MAI1 has 10 standards, while the software has only 6 standards and each one has a score, the total score is MAI per patient. Results: in this project 1848 patients were included, at admissions they were classified frail using the Multidimensional Prognostic | @EPSA_Online

Questions & answers

whilst carrying out the research and how did you overcome that? The biggest challenge was working in a team with such a wide background: I have never had to work with clinicians on therapies before my research project. Research on this project has been opened since 2012, and all doctors in the ward have had many years of experience in clinical practice, one of them was even a university professor as well. When I first arrived in the ward, I had to do a lot of research to improve my knowledge on the matter and my skills for my final discussion, but was also curious about clinical practices and I made myself always available to do tasks outside my day to day job, so I could integrate well in the team. If there were more complex clinical situations, I searched through online resources for possible relationships between drugs and hospitalisations.

Please tell us a little bit more about yourself. II am Martina Tammaro, I am Italian and I have recently graduated in pharmacy - December 2016 - from Padua university. In June, I also passed my license to practise as a pharmacist and now I am searching for a job opportunity and I am really interested in hospital pharmacy and in the role that a pharmacist can have in the Health Systems, so I hope my career path will follow that direction. I do not know EPSA very well but I am interested and would like to In your opinion, what is the benefit know more about it. of joining ESSP and what advice do Tell us a bit more about your research you have for students undertaking research in the future? and its significance. This research is part of my experimental It is an honour to be published, and having thesis, it took me 7 months to complete and the possibility to learn about the work of other during this period I had the chance to stay in a European research teams. It is an experience Geriatric ward. It was an amazing experience: because it has improved a lot my skills, such in Italy there is not usually a pharmacist in the as writing an abstract or respecting the steps hospital ward, even if in many other countries for the publication. Furthermore if English is this is common. In this way, I was able to learn not your first language, you have the possibility to improve your English. some more clinical matters. During university I studied a lot of pharmacology and pharmacovigilance but I have not yet had the possibility to use my knowledge in real life situations; during my stay in the geriatric ward, I finally got to see the application of my studies. The research was very important because Geriatric patients are usually excluded from clinical trials and yet they are the biggest drug consumers and they are a significant presence in polypharmacy, although there is still no awareness, in clinical practice, of drug interactions. What





In this crisis period I think it is very important to create links between other countries and other research projects and I hope that our researches can be carry out across Europe. I think that in Italy there is a very important professional crisis which pharmacists can solve only in an European context.


European Pharmaceutical Students’ Association

The role of a pharmacist in the pharmacovigilance system Author(s): Marija Lucic (, Nastasija Milosevic, Aleksandar Raskovic Institution: Faculty of Medicine, University of Novi Sad, Serbia

Introduction: Although they represent an important mediator between patients and the National Pharmacovigilance Centre, pharmacists still do not participate enough in the system of reporting adverse drug reactions (ADR). By reporting ADR, both quality of therapy and quality of patient’s life improve, and the pharmaceutical industry is also encouraged to invent and produce new formulations which will be better accepted and whose use will significantly improve the risk-benefit ratio. Aim: The aim is that by collecting information about unexpected and adverse drug reactions, based on direct contact of pharmacists and patients, we will improve the role of a pharmacist in the pharmacovigilance system.

of seriousness.

Conclusion: Thanks to direct communication between a pharmacist and patients and additional information about the importance of reporting unexpected and adverse reactions, both the role of a Material and Methods: Information about pharmacist in pharmacovigilance system and adverse reactions were being collected in safety of pharmacotherapy can be improved. pharmacies in Inđija and in Sombor. Between the period of 20.12.2017.-10.01.2018., Keywords: unexpected and adverse effects; pharmacists were taking notes about pharmacovigilance. adverse reactions of medicines. Between the period of 11.01.-01.02.2018. patients were additionally informed about unexpected and adverse drug reactions. The standard form for reporting an adverse reaction was filled for each reported case and sent to the National Pharmacovigilance Center. Results: During the first period, without direct involvement of pharmacists, there were 19 reported adverse reactions. After that, when patients were additionally informed about the importance of adverse effects, there were 33 reported cases. The most numerous adverse drug effects were from group C, according to ATC classification, or cardiovascular drugs (32,7%), then, from group J or anti-infective drugs with systemic effects (15,4%) and from group N or drugs which affect the nervous system. After analysing reported reactions, according to NPC, all of the reactions were expected (52), but 3 of them fulfilled criterion | @EPSA_Online

Questions & answers

reactions they experienced. I tried to point out to them how important this is, and explain that, by reporting, they can save somebody’s life. A lot of them did not know that a system who notes all the reported reactions exists, so I had to explain how all that functions to them. In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? ESSP gives all of us the opportunity to make contacts with people who share the same interests as we do by publishing our abstracts. Also, by reading other students’ abstracts we can learn something new and keep up with news in the field of pharmacy.

Please, tell us a little bit more about yourself. My name is Marija Lucic, and I am a 4th year student at Pharmacy and Medical Faculty in Novi Sad, Serbia. I enjoy attending all the events organised by our local students’ organisation (PSANS-Pharmaceutical Students’ Association of Novi Sad) and national students’ organisation (NAPSerNational Association of Pharmacy Students I strongly recommend to all students to undertake research during their studies - Serbia) and taking parts in other activities. because it gives you a whole new experience. That is the way I fulfill my knowledge and You have to have patience, to be stubborn make amazing friendships! I am also very and positive, and everything will turn out great. passionate and curious when pharmacology Remember that hard work always pay off! comes to the question. I am especially interested in studying drug interactions and adverse reactions and that is the reason I chose pharmacovigilance as the topic of my study. Tell us a bit more about your research and its significance. The aim of this study was to see and to improve the role of pharmacists in the pharmacovigilance system in Serbia. Reporting adverse drug reactions is important, and medical workers are not aware of it enough. Pharmacists spend a lot of time talking about drug consumption with patients, so they should also inform them about the importance of reporting unexpected and adverse drug reactions. What was the biggest challenge whilst carrying out the research and how did you overcome that? The biggest challenge for me was to interest patients in reporting information about adverse


European Pharmaceutical Students’ Association

The identification of the genetic cause of precocious puberty Authors: Kaja Mivšek, izr. prof. dr. Katarina Trebušak Podkrajšek, univ. dipl. kem., spec. med. biokem., spec. lab. med. gen., doc. dr. Magdalena Avbelj Stefanija, dr. med., dr. Jernej Kovač, uni. dipl. biokem. 1. Clinic of Pediatrics – University Children’s Hospital, Ljubljana University Medical Centre, Unit of Special Laboratory Diagnostics, Vrazov trg 1, Ljubljana, Slovenia 2. Faculty of Pharmacy, Aškerčeva cesta 7, Ljubljana, Slovenia

Introduction: Interactions between genetic and environmental factors determine the timing of sexual maturation. Puberty begins when the hypothalamic-pituitary-gonadal axis is reactivated. At that time, mammary glands in girls and testes in boys are developed, and growth and bone maturation are accelerated. When this pubertal characteristics occur before the expected age, possible cause for them is central precocious puberty. Premature pubertal development results in the shift of a fertile period, which is associated with the risk of developing certain diseases, a greater tendency towards the lower final adult stature and increased potential for psychosociological disorders. Molecular genetic testing is needed to identify the potential hereditary cause of the disease. Aim: The aim of this study was to detect genetic variants that could be responsible for precocious puberty. We determined the sequence of the entire genome in members of four families with maternally inherited precocious puberty. We applied the whole genome sequencing followed by bioinformatics approaches in order to identify the possible genetic causes of precocious puberty. Materials and Methods: Using tools and algorithms, we identified variants in all four families located in the same gene ZNF717 and therefore determined them as potentially pathogenic. We quantitatively assessed the pathogenicity of these variants. In members of the fourth family, we verified the presence of the most likely pathogenic variant by confirmative Sanger sequencing. Results of Sanger analysis showed that the desired variant was not present, indicating error in the approach of whole genome sequencing and subsequent bioinformatics analysis. The causes of the false positive result were discussed and the weaknesses of the whole genome sequencing method and the bioinformatics analysis were presented.

Results: The results of the initial analyses were not reliable, therefore potentially causative genetic variants in analysed families with precocious puberty remain undetermined. Our results showed that verification of the results of next generation sequencing with a secondary method is necessary for reliable genetic diagnostics. | @EPSA_Online

using computer tools. Unfortunately, these tools were most of the time not efficient enough to process this much data at once. For this reason and due to the limited time for which I was available for the research of my Master’s degree, we could not examine the entire genomes to identify possible causative genetic mutations, so we only checked coding regions. The study is going to continue Please, tell us a little bit more about on a new and more powerful computer with which we are going to analyse the remaining yourself. My name is Kaja Mivšek, I’m 26 years old and genomic regions! We hope to find the potential hereditary cause of the disease. I’m from Vrhnika, Slovenia!

Questions & answers

I started my studies at the Biotechnical Faculty, University of Ljubljana (bachelor study programme Biology – 3 years) and continued at the Faculty of Pharmacy, University of Ljubljana (master study programme Laboratory Medicine – 2 years). I successfully finished both study programmes!

Another thing that surprised us were falsepositive results. Identifying causes that could lead to this rare event presented a challenge, since they are more comprehensive as they seem. We found several different possibilities that could have caused such results.

In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? The most important benefit of ESSP Tell us a bit more about your research participation for me is to become familiar with the process of publishing research results and and its significance. This research was part of an international to receive professional feedback on writing project of Pediatric Endocrinology (ESPE articles. Research Unit Grant), which aims to identify new genetic causes of the familial My advice to all of you, who are going to precocious puberty. We were searching perform scientific research and publish it, for possible genetic causes of precocious is to read as much as possible about the puberty, in whole genomes of members of 4 problem of the study before you actually begin families with maternally inherited precocious with practical work. Read all of the scientific puberty. In order to identify them, we applied papers that have been written on your topic the whole genome sequencing, followed before you begin with yours. This is how you by bioinformatic approaches. Finally, we will get to know how far and deep science performed a confirmatory test. I used the actually already is with studying your topic. It is also very important that the aim of the results to write my Master’s Thesis. study is fully clarified to you. First, perform What was the biggest challenge the practical part of the research and (this is whilst carrying out the research and important) attentively write down every single detail you come across at your work. After the how did you overcome that? The biggest problem was the huge amount practical part is completed, do not hesitate to of data we acquired by sequencing the whole start writing the actual scientific paper and do human genome. We had to analyse this data not stop until you are done! In my spare time I really like reading inspirational and scientific stuff, being in nature, running, watching documentaries, listening to music, among others!


European Pharmaceutical Students’ Association

Comparison between the nitrogen oxides inhaled when using regular cigarettes versus IQOS cigarettes (HEETS) Student: Ananda Diana Pop, Nicoleta Nistorescu Coordinator: Associate Professor Mircea Dumitru Croitoru Department of toxicology, University of Medicine and Pharmacy Targu Mures, Romania

Introduction: Smoking has been proven to be a significant health hazard and a leading cause of malignant tumor formation. Nowadays, methods to reduce health risks associated with smoking are appearing on the market, such as electronic cigarettes and the IQOS system. The IQOS system is promoted as a safer way to smoke than the regular cigarettes due to the fact that it eliminates the process of burning. Aim: In the present work we measured the amounts of nitrogen oxides inhaled by a smoker using regular cigarettes versus using the IQOS system. Determinations were made on three types of cigarettes and heets: weak, medium and strong. Material and Methods: Smoke resulting from using the two smoking devices was absorbed. We used a sodium hydroxide solution as an absorber of the cigarette smoke which was produced by means of a vacuum pump adjusted, so as to mimic the normal smoking as closely as possible. Measurement of nitrogen oxides was made by using a diazotisation method available in the scientific literature. The measurement of coloured substances was made by reading the absorbance of the solution at 520 nm. We took the working method from the Practice Toxicology book named “Ghid practic de analiza a toxicilor gazosi si volatili by Mircea D. Croitoru” after checking it on the ScienceDirect site: (https://www. griess-test). Results: Significant differences were observed regarding the presence of nitrogen oxides in the smoke between the two ways of smoking. Six cigarettes were measured from each package. A total of 36 cigarettes were

measured once. There was no significant difference between hard, medium and weak cigarettes. Regular cigarettes produced between 7-37 ug nitrogen dioxide/cigarette while in the case of the IQOS system, the amount of nitrogen dioxide that was formed fell under the limit of detection. A 10 times decrease was observed in the case of the IQOS system. The solution from IQOS was much more discolored, so extinctions were statistically significant (p>0.0001). By comparing the average extinctions from the table for the two types of cigarettes, you can see an about 10 times decrease for Heets. By performing the T test (Student’s t-test) for the extinctions obtained on the two types of cigarettes, we can state that the results were statistically significant. Measurement of nitrogen oxides was made by using a diazotisation method. In the case of normal cigarettes, the amount of nitrogen oxides could be measured. Heets cigarettes were below the limit of detection. | @EPSA_Online

*This is the average and standard deviation byproducts resulting from burning, and not of a simple extinction for normal cigarettes. necessarily nicotine. (Extinction/cigarette ) Efforts are being made to eliminate the *This is the average and the standard deviation burning process and to deliver nicotine in of the simple extinctions for heets cigarettes. another form. Based on the results obtained, (Extinction/HEETS) we want to inform the public about the risks they pose due to smoking and the significant Conclusion: Our results show that differences that may occur between normal significant reduction in the formation of toxic smoking or the IQOS device. substances can be achieved by using the IQOS system compared with regular smoking. We measured spectrophotometric nitrogen Measurement of other toxic species should oxides at 520 nm wavelength. We obtained be made in order to obtain clearer information the absorbance of the solution and assumed about the health risk differences between the it was related to the amount of combustion two smoking approaches. products that resulted. In the future we want to determine other compounds in cigarettes and the cigarette smoke which can affect our health. What was the biggest challenge whilst carrying out the research and how did you overcome that? The biggest challenge we encountered during the experiment was procuring a cigarette Please, tell us a little bit more about smoke-absorbing device. yourself. I, Ananda Pop, and my colleague, Nicoleta We used an absorber, whereby in the alkaline Nistorescu are students in the 4th year at UMF solution (NaOH) we absorbed the cigarette Targu Mures. In high school we discovered smoke by means of an adjustable vacuum our passion for chemistry and we decided to pump. We tried to mimic the normal smoke pursue a career in pharmacy. as closely as possible.

Questions & answers

Our favorite motto is “Our prime purpose in life is to help others� by Dalai Lama. Supported by the desire to help other people, we started this research when we found out that a new type of electronic cigarettes appeared on the market and knowing the risks associated to being a smoker, we started studying the subject in detail.

In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? There are many benefits that come with ESSP. One of them would be to interact with people in the same field as you, but from different locations. Another advantage is the possibility for a student to publish a scientific paper and Tell us a bit more about your research to assert yourself in the field in which you have and its significance. chosen to practice. The main purpose of this scientific paper was to determine the difference in nitrogen oxides My advice to students is never to give up and resulting from cigarette smoke. Problems always visualise the ultimate goal! arise due to the burning of cigarettes and


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Fabrication of a biodegradable nanofiber loaded with lidocaine for hernia repair surgery Author: Beatriz Santos Mentor: Dr. Greg Walker, Prof. Gerrit Borchard School of Pharmacy, Institution: University of Otago, New Zealand and Université de Genève, Switzerland

Introduction: Hernia repair using mesh has become a standard procedure in most countries. Local anaesthetics (LAs) such as lidocaine have proven to be efficient in alleviating postoperative symptoms, thus finding a drug delivery system that allows the administration of lidocaine locally, would help reducing convalescence and improving recovery. Aim: The goal of this project was to electrospin a lidocaine-loaded nanofiber that could be used as a complement to polypropylene meshes in a hernia repair surgery. A study of the nanofiber properties, as well as the capacity of entrapment and release of the drug, is performed. Material and Methods: Poly (lactic-coglycolic acid) solutions with a lactide:glycolide ratios of 50:50 and 75:25, were prepared in Acetone/Dimethylformamide in ratio 80:20. Lidocaine was added to the polymer solution and electrospun at different flow rates and voltages. The fibres mats were characterised using SEM. FTIR was used to determine chemical changes to the polymer. The mechanical strength was measured using a tensile strength apparatus. Degradation of the mats was evaluated in vitro by measuring mass loss over ten weeks. HPLC analysis of lidocaine was performed in an isocratic mode. The entrapment efficiency was evaluated by dissolving the mat in the solvent system and analysing the lidocaine content by HPLC. A preliminary drug release study was performed with samples in PBS for a seven days period using HPLC. Results: PLGA nanofibers containing lidocaine were electrospun using Acetone/ Dimethylformamide (80:20) as the less toxic solvent system. SEM showed PLGA 75:25 fibres to be more coalescent as the amount of

drug loading increased. No major difference was observed in the morphology of PLGA 50:50. FTIR spectra show no shift changes with increased drug loading. For both PLGA ratios nanofiber mats, tensile strength decreases with the increasing amount of drug loaded. The degradation studies showed a slight difference in the degradation rate for PLGA 50:50 and 75:25 and for both, a 50% mass loss after ten weeks. The entrapment efficiency was not lower than 60% for both ratios. Preliminary drug release studies showed nearly 100% drug released after only twelve hours. The final release study of the drug is still in progress. Conclusion: These studies demonstrated that lidocaine can be successfully entrapped into a PLGA 50:50 or 75:25 nanofiber mats and this new formulation may be suitable to be applied to the body with a hernia mesh for the local delivery of lidocaine for pain relief. | @EPSA_Online

benefit of joining ESSP. We are the present and future and the world is in our hands. It is great to see motivated people, who believe in what they do.

Questions & answers

For all of you undertaking research in the future, do what you love, regardless of what you may hear. If you believe in it, than it is already worth it.

Please, tell us a little bit more about And remember: “Everything will be okay in the yourself. My name is Beatriz and I am a 24 year old end. If it’s not okay, it’s not the end”. portuguese studying in Switzerland. I am constantly looking for new challenges and believe that life would not be as thriving without a little failure. I ask a lot of questions but do not have half the answers. Definitely, there are still amazing things to be done in the health field, and I hope I can contribute. What was the biggest challenge whilst carrying out the research and how did you overcome that? I did my research project 18’000 km away from home and this was clearly the biggest challenge for me. New Zealand is an amazing country filled with incredible people who do not always remember to pay the electric bill in the middle of winter! When I was told I was to make a drug loaded nanofiber to be used in hernia repair surgery, I had no idea where to start. Up until then, I always had a protocol, people to tell me what to do and so, realizing that I was »on my own« scared me. This clearly made me more confident and helped me grow! In your opinion, what is the benefit of joining ESSP and what advice do you have for students undertaking research in the future? Being able to share your work with people you have never met is, in my opinion, the biggest


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Microfluidics for polymer encapsulation of drug-loaded mesoporous silica nanoparticles (MSN) Author: Ibrahim Mamudu,( Supervisor: Asist.Prof.Hongbo Zhang Institution: Pharmaceutical Sciences Laboratory, Åbo Akademi University, Finland

Introduction: Mesoporous silica nanoparticles (MSN) are of significant interest for therapeutic applications in recent years due to their stability, biocompatibility and high drug loading properties. In targeted delivery systems, nanoparticles are encapsulated with polymers such as acetylated dextran (Ac-Dex) which hydrolyses in acidic conditions, but remains stable in neutral environment. However, the encapsulation of nanoparticles has been a major challenge due to their extremely small size and high surface area. In this study, drug-loaded small positively charged mesoporous nanoparticles (S-MSN(+)) were encapsulated with Ac-Dex using microfluidics. Microfluidics is capable of handling small liquid quantities and therefore has proven useful in nanoparticle encapsulation. Aim: Use microfluidics to fabricate AcDex-coated S-MSN(+) nanocomposites (S-MSN(+)@Ac-Dex).To investigate the drug loading capacity (DLC) of bare S-MSN(+) and encapsulated S-MSN@Ac-Dex nanocomposites. Material and Methods: Two samples of drug-loaded S-MSN(+) were prepared. One sample was used to evaluate the drug loading capacity of bare S-MSN(+), while the other to perform Ac-Dex encapsulation to produce S-MSN(+)@Ac-Dex nanocomposites using a 3D co-flow glass capillary microfluidic device. The formed bare S-MSN(+) and S-MSN(+)@ Ac-Dex were evaluated with Zetasizer Nano ZS to determine size, polydispersity index (PDI) and zeta potential. The two samples were then shaken for 12 hours in dimethyl sulfoxide (DMSO) to unload the drug. UV absorption of the released drug was measured using Nanodrop 2000c spectrophotometer at 254 nm and the corresponding concentrations were obtained using standard curve for the calculation of DLC. The drug A-769662 (CAS number: 844499-71-4) used was a lung cancer inhibitor testing drug. Results: Bare S-MSN(+) had a particle

size of 137.3±0.7 nm, a PDI of 0.031±0.01 and a zeta potential of +37.3±1.5 mV while S-MSN(+)@Ac-Dex had a particle size of 259±1.2 nm, a PDI of 0.264±0.006 and a zeta potential of -26.6±1.2 mV. The DLC of bare S-MSN(+) and S-MSN(+)@Ac-Dex was 17.4% and 28.6%, respectively. Conclusion: The change in zeta potential of bare S-MSN(+) from +37.3 mV to -26.6 mV after Ac-Dex encapsulation shows coating was successful. Fabrication of S-MSN(+)@ Ac-Dex nanocomposites with DLC of 28.6% using microfluidics was demonstrated. However, this high DLC value of S-MSN(+)@ Ac-Dex could possibly be incorrect due to interference in UV absorption of the drug at 254 nm by the presence of Ac-Dex. Therefore, further investigation using high pressure liquid chromatography (HPLC) is required to measure the actual DLC of S-MSN(+)@AcDex. | @EPSA_Online

Questions & answers Please, tell us a little bit more about yourself. My name is Ibrahim Mamudu from Ghana (West Africa) and I am 22 years old. I am a 4th year international student in Hacettepe University Department of Pharmacy (Bachelors), Turkey. Tell us a bit more about your research and its significance. In recent years,various techniques have been developed to produce encapsulated nanodrugs to target cancer cells. However, fabrication of these nanodrugs with desirable drug loading capacity is a major challenge due to the small nature and high surface area of nanoparticles. To address this problem, we used a microfluidic technique to successfully fabricate nanodrugs which have considerably high drug-loading capacity. This goes to show that, microfluidics could become the new paradigm for nanomedicine formulation and production for cancer treatment in the future. What was the biggest challenge whilst carrying out the research and how did you overcome that? I had to read a lot of research papers to familiarise myself with the topic, because I did not study the topic at my university. Another challenge I faced was the fact that I only had 2 months to finish the project, so I had to spend a lot of hours working in the laboratory, but i did not give up! In your opinion, what is the benefit of joining ESSP and what advice do

you have for students undertaking research in the future? ESSP has provided with me with an opportunity to develop skills in writing abstracts and also to share a topic I am passionate about, with other pharmacy students. I advise students not to lose their enthusiasm when searching for research opportunities in the field of their interest, even after facing several rejections! Also, it is essential to know that professors and mentors are there to guide you, so have the courage to pursue a new research.

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