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education and training of the pharmacists

It was demonstrated that nanobodies with an in vitro neutralizing activity against poliovirus type 1 interact with native virions.Here, the use of capillary electrophoresis was investigated as an alternative technique for the evaluation of the formation of nanobody-poliovirus complexes, and therefore predicting the in vitro neutralizing activity of the nanobodies. The macromolecules are preincubated offline in a specific nanobody-to-virus ratio and analyzed by capillary electrophoresis with UV detection. At low nanobody-to-virus ratios, a clear shift in migration time of the viral peak was observed. A broad peak was obtained, indicating the presence of a heterogeneous population of nanobody-virion complexes, caused by the binding of different numbers of nanobodies to the virus particle. At elevated nanobody-tovirus ratios, a cluster of peaks appeared, showing an additional increase inmigration times. It was shown that, at these high molar excesses, aggregates were formed. The developed capillary electrophoresismethod can be used as a rapid, qualitative screening for the affinity between poliovirus and nanobodies, based on a clearly visible and measurable shift in migration time. The advantages of this technique include that there is no need for antigen immobilization as in enzyme-linked immunosorbent assays or surface plasmon resonance for the use of radiolabeled virus or for the performance of labor- and time-intensive plaqueforming neutralization assays.

Previously, we reported on the in vitro antiviral activity of five single-domain antibody fragments (VHHs) directed against poliovirus type 1. In the present study, we further investigated the mechanism of action of these VHHs, using both cell-based assays and cryo electron microsocopy.

Previously, we produced and selected single-domain antibody fragments (VHHs) with an in vitro antiviral activity against poliovirus type 1. In the present study, we further investigated the mechanism of action of these VHHs, using both cell-based assays and cryo electron microsocopy.

Previously, we reported on the in vitro antiviral activity of single-domain antibody fragments (VHHs) directed against poliovirus type 1. Five VHHs were found to neutralize poliovirus type 1 in an in vitro setting and showed 50% effective concentrations in the nanomolar range. In the present study we further investigated the mechanism of action of these VHHs. All five VHHs interfere at multiple levels of the viral replication cycle, as they interfere both with attachment of the virus to cells and with viral uncoating. The latter effect is consistent with their ability to stabilize the poliovirus capsid, as observed ina thermo-fluor thermal shift assay, in which the virus is gradually heated and the temperature causing 50% of the RNA to be released from the capsid is determined, either in the presence of in the absence of the VHHs. The VHH-capsid interactions were also seen to induce aggregation of the virus-VHH complexes. However, this observation cannot yet be linked to their mechanism of action. Cryo-electron microscopy (cryo-EM) reconstructions of two VHHs in complex with poliovirus type 1 show no conformational changes of the capsid to explain this aggregation. On the other hand, these reconstructions do show that the binding sites of VHHs PVSP6A and PVSP29F overlap the binding site for the poliovirus receptor (CD155/PVR) and span interfaces that are altered during receptor-induced conformational changes associated with cell entry. This may explain the interference at the level of cell attachment of the virus as well as their effect on uncoating.

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The PHAR-QA by PHARMINE

The European Union directive on sectoral professions emphasizes the fact that pharmacists working in member states should possess the competences required for their professional practice; the directive does not, however, describe such competences in detail. The "Quality Assurance in European Pharmacy Education and Training--PHAR-QA" consortium, funded by the European Union, will define such competences and establish a quality assurance system based on them. This will facilitate the tuning of the pharmacy education and training required to produce competent pharmacists in the different member states. PHAR-QA will (1) establish a network of participating pharmacy departments, (2) survey existing quality assurance systems used, and (3) develop competences through iterative interaction with partners. The European Association of Faculties of Pharmacy will use the harmonized competences produced as a basis for the creation of a quality assurance agency for European pharmacy education and training. PHAR-QA will impact on staff and students of European departments; the final stake-holder will be the European patient who will benefit from better pharmaceutical services and better medications.

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during the first year of the bachelor pharmacy students start with the course "line project I". The next three years of the bachelor program they acquire and expand their competences. Reflection, as self regulation or the possibility to improve the own action, is important in this learning process.

The PHAR-IN consortium (538252-LLP-1-2013-1-BE-ERASMUS-EKA) consists of pharmacy faculties and industrial partners from countries of the European Higher Education Area (EHEA), members of the European Association of Faculties of Pharmacy (EAFP) together with a professional organisation representing industrial pharmacists, the European Industrial Pharmacists's Group (EIPG). PHAR-IN is funded by the Eur Comm (EC) via its education, audio-visual and culture agency (EACEA).

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Health professionals at the helm of change for self-care

Background: In the Master program of Medicine at the Vrije Universiteit Brussel, the Brussels Clinical Evaluation (BRUCE) serves as a monthly student based evaluation instrument of the educational performance of clinical professors and departments. BRUCE started out as BRUCE 1.0 (paper version) and was used from February 2010 until June 2011. Summary of work: Before introducing an electronic version, BRUCE 2.0, a qualitative and quantitative analysis of BRUCE 1.0 was done. The present study correlates both. Summary of results: Two different situations appeared. On one hand there are questions with a high level of missing values and a lower mean and negative qualitative scores on relevance and clarity. On the other hand there are questions with a high level of missing values and a lower mean and no negative qualitative scores. Discussion: Comparison of qualitative and quantitative study reveals why certain items have a high level of missing values and a low mean. The first situation reveals a construction flaw in the questions themselves, while the second situation reveals an effective qualitative problem within the internship. Conclusion: Qualitative analysis helps to understand reasons for missing values and lower means. Take home messages: When validating an instrument combining quantitative analysis with qualitative analysis is indispensable.

Editorial

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Nanobodies or single domain antibodies are the smallest antigen-recognizing parts of the heavy-chain-antibodies found in camels and other members of the Camelidae. They are known to be very stable, specific and non-immunogenic and are therefore promising candidates to be developed as an anti-polioviral drug. We previously reported about a set of nanobodies that show an in vitro neutralizing activity on poliovirus type 1. Due to the generation of neutralization-escape virus mutants during treatment of infected HeLa cells with these nanobodies, a combination therapy with two or more nanobodies targeting different epitopes would be essential to overcome the induction of drug-resistant virus. Nanobodies are encoded by single-genes and can be easily tailored in different formats like bispecific (two different nanobodies) or bivalent (two identical nanobodies) constructs. These constructs are expected to show a higher avidity for their antigen as they possess two paratopes. Here, we compare the influence on the in vitro neutralizing activity of five different protein-linkers, differing from each other in both length and flexibility, by making constructs of nanobodies PVSP29F and PVSP6A (known to target two different epitopes).

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Nanobodies are recombinant single-domain antibody fragments deriving from heavy-chain antibodies (HCAb) found in camelids. Since the antigen-binding site of the camelid HCAb is comprised in one single domain, the nanobody harbors the full antigen-binding capacity of naturally occurring HCAb. Because of their characteristic structure, nanobodies have the tendency to bind into cavities. In this study the epitopes of five neutralizing nanobodies against poliovirus type 1 were mapped on the deduced amino acid sequence of the capsid coding region from viral neutralizing-escape variants.

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Despite the existence of effective vaccines and the numerous vaccination campaigns worldwide, almost 1000 new cases of poliomyelitis are reported each year by the World Health Organization. The causative agent of this paralytic disease is poliovirus, a member of the family of the Picornaviridae. The eradication of this virus is tougher than expected at first and the need for an antiviral drug to support vaccination in the eradication and post-eradication era has become more and more evident . Nanobodies seem to be perfect candidates to be developed as an anti-polioviral drug. Nanobodies are single domain antibodies derived from the heavy chain antibodies found in camelids and show some very interesting characteristics as they combine the advantages of conventional antibody therapeutics (i.e., high target specificity, high target affinity and low inherent toxicity) with the key features of small-molecule drugs (e.g. recognition of hidden or uncommon epitopes, binding into cavities, drug format flexibility, gastro-intestinal stability, tissue penetration, tailoring of half-life...) . We developed a set of nanobodies against poliovirus strain Sabin 1 and we tested their poliovirus-neutralizing activity by measuring the reduction in cytopathic effect induced by poliovirus on HeLa cells treated with the nanobodies. Twelve nanobodies were found to neutralize the vaccine and wildtype strain of poliovirus type 1. We have shown that different nanobodies target different epitopes on the virus capsid and in terms of the development of therapy-resistance, it would be interesting to use a combination of nanobodies to neutralize the virus. A possible way of combining them is to create contructs of two or more poliovirus-neutralizing nanobodies recognizing different epitopes. These constructs are easily made by linking the genes coding for two nanobodies to each other with a linker/spacer sequence. Like this the construct is coded by one gene and can be produced during one single production process. Due to their dimensions, theconstructs have an increased half-life value, which is an advantage in terms of therapy adherence.

There is no discussion that there is an evolution, even a revolution going on, and still coming up, in the pharmacy profession. For many years, the pharmacist was responsible for the production and the analysis of medicines. Now, the pharmacist is also responsible for the information towards patients and the profession is now more than ever patient-centered than medicine-centered. This evolution had also structural influences on pharmacy education. Which new topics should be included in the curricula? Which new educational methods should be introduced? How to find competent teachers? Moreover, whereas pharmacy curricula were originally focused on "hard sciences", such as chemistry, physics, mathematics, more so-called "soft sciences" have to be introduced in the pharmacy programme. Examples of these soft sciences are communication (patient-related, but also communication with other health care workers), pharmaceutical care and social pharmacy. These changes in pharmacy education have an important influence on management issues in the Schools of Pharmacy. Most of the research in these so-called soft sciences is more difficult to finance (less publications, journals with lower impact factor, etc.). Also the clash between the traditional faculty (hard sciences) and the new faculty(soft sciences) is difficult to avoid. This is definitely a challenge for managers and Deans of Schools of Pharmacy in a global wide perspective. Finally, we may not forget that the pharmacy programme should be above all an academic programme, this to distinguish pharmacy (and other sectorial professions) from the main stream of professional education.

In the master programme of medicine at the VUB, the internships play an important role. In 2009-2010, BRUCE 1.0 was developed as a new student based evoluation instrument of the educational performance of clinical professors and the clerkship rotations in general, this in close dialogue with the students.

This paper results from the poster exhibition of the international conference on the Structure of Medical Education in Europe, hosted by the German Rector's Conference (HRK), which took place on 10/11 October 2008 in Berlin. In response to the European Bologna guidelines to create a European Higher Education Area in which students should become more mobile and to the need of more flexibility in higher education also, the Vrije Universiteit Brussel (VUB) installed in 2004 'Academic Career Consultants' (ACCs) open to all (potential) students, national and international. The ACC deals with questions or challenges in connection with the (flexible and/or mobile) academic educational path. For the students (wanting to study at) the Faculty of Medicine and Pharmacy of the VUB there is a full time ACC. From the five years of experience in working with medical students VUB wants to inform all involved medical education professionals on the assets of academic career consultancy. VUB also wants to invite others to help setting up a quality driven international network of ACCs.

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The Bologna declaration and the EU directive 2005/36/EC on the recognition of professional qualifications influence the mobility of pharmacy students and pharmacy professionals.

Interviewed by Sanziana Marcu-Lapadat, EPSA Vice President of External Affairs

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Round table conversation

Recently, single domain recombinant antibody fragments (VHHs or Nanobodies) against poliovirus were isolated from a dromedary immunized with poliovirus type 1 Sabin strain. In this study, the interaction of a panel of fifteen VHHs with native infectious poliovirus (N antigen), heat-denatured virus (H antigen) and 14S subviral particles was examined in a quantitative affinity capture assay. Five distinct subsets of VHHs are tentatively distinguished, of which at least two bind to epitopes not previously reported. Furthermore, we demonstrate that all neutralizing VHHs recognize N antigen and all N-specific VHHs are neutralizing. We suggest that neutralizing VHHs can be used as an additional antiviral tool compatible with the protective immune response of vaccination to control poliovirus circulation or future poliovirus outbreaks. To summarize, this study corroborates the findings that VHHs target mainly conformational epitopes and target other epitopes than classical antibodies. This technique can be used for epitope mapping and tracking conformational changes of proteins.

In the 21st century, community, and also hospital pharmacists, will play an increasingly important role as partners in the efficient use of the health care resources of the EU.

The roles and responsibilities of the modern-day pharmacist are evolving very quickly, and pharmacy education and training will have to adapt in order to provide the competences needed for the new roles and responsibilities.

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Poster presented at EUROPIC 2010 conference, September 12th, 2010, St. Andrews.

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PHARMINE (pharmaceutical education in Europe), a challenge to pharmacists

The aims of the PHARMINE consortium