Novel 5-aryloxypyrimidine SEN1576 as a candidate for the treatment of Alzheimer's disease

Prefibrillar assembly of amyloid-ß (Aß) is a major event underlying the development of neuropathology and dementia in Alzheimer's disease (AD). This study determined the neuroprotective properties of an orally bioavailable Aß synaptotoxicity inhibitor, SEN1576. Binding of SEN1576 to monomeric Aß 1–42 was measured using surface plasmon resonance. Thioflavin-T and MTT assays determined the ability of SEN1576 to block Aß 1–42-induced aggregation and reduction in cell viability, respectively. In vivo long-term potentiation (LTP) determined effects on synaptic toxicity induced by intracerebroventricular (i.c.v.) injection of cell-derived Aß oligomers. An operant behavioural schedule measured effects of oral administration following i.c.v. injection of Aß oligomers in normal rats. SEN1576 bound to monomeric Aß 1–42, protected neuronal cells exposed to Aß 1–42, reduced deficits in in vivo LTP and behaviour. SEN1576 exhibits the necessary features of a drug candidate for further development as a disease modifying treatment for the early stages of AD-like dementia.

The student experience in Objective Structured Clinical Examinations (OSCEs): phenomenological analysis

Background and purpose
The dominant psychometric discourse of OSCEs may lead to unexpected problems, such as a checklist-based student performance1 which under emphasises the clinical relationship with student and standardised patient (SP). Such encounters can be dehumanising for SPs2 and have implications for what students learn about relational skills through the assessment process. In this study we explore medical students’ experiences of undertaking OSCEs using a phenomenological frame.
Methodology
Interpretative phenomenological analysis is a form of qualitative methodology which has strong resonance with existentialism and focuses on the lived experience without significant reference to external political or discursive
forces.
Six 4th year undergraduate medical students from Queen’s University Belfast were recruited in December 2013. Maximum variation sampling was used. Students were interviewed by a researcher in the week prior to the
OSCE and then again in the week following the OSCE in Jan 2014. Interviews were minimally structured in order to be open to respondents, rather than adhering to a fixed topic guide, but focussed on participants’ experiences, thoughts and feelings about taking part in OSCEs. Interviews were audio-recorded and
transcribed. Students were also asked to complete a short diary entry in the days prior to the OSCEs and another immediately following. Diary entries were written, emailed or audio-recorded at student’s preference.
Results
Transcripts are currently being analysed by interpretative phenomenological analysis. Preliminary analysis has demonstrated the significance of students’ relationships within the OSCE triad (student, SP and examiner); the effect of the immediate examination environment; realism versus roleplay; students’ perceptions of the purpose of assessment; and coping mechanisms.
Full results will be available by the time of the conference.
Conclusion and Discussion
Understanding the student experience in OSCEs is a crucial step in understanding the complex construction of relationships within the OSCE triad. The focus in OSCEs is typically on standardisation and reliability, but in exploring social interactions we may refocus attention on their inherent potential for learning and effects on both students and patients.
References
1. Hodges B. Medical education and the maintenance of incompetence. Med Teach 2006;28(8):690-6
2. Johnston JL, Lundy G, McCullough M, Gormley GJ. The view from over there: reframing the OSCE through the experience of standardised patient
raters. Med Educ 2013;47(9):899-909

Genetic studies of body mass index yield new insights for obesity biology

Obesity is heritable and predisposes to many diseases. To understand the genetic basis of obesity better, here we conduct a genome-wide association study and Metabochip meta-analysis of body mass index (BMI), a measure commonly used to define obesity and assess adiposity, in up to 339,224 individuals. This analysis identifies 97 BMI-associated loci (P < 5 × 10(-8)), 56 of which are novel. Five loci demonstrate clear evidence of several independent association signals, and many loci have significant effects on other metabolic phenotypes. The 97 loci account for ∼2.7% of BMI variation, and genome-wide estimates suggest that common variation accounts for >20% of BMI variation. Pathway analyses provide strong support for a role of the central nervous system in obesity susceptibility and implicate new genes and pathways, including those related to synaptic function, glutamate signalling, insulin secretion/action, energy metabolism, lipid biology and adipogenesis.

The emergence of DNA methylation as a key modulator of aberrant cell death in prostate cancer

It is now well established that cancer cells exhibit a number of genetic defects in the machinery that governs programmed cell death and that sabotage of apoptosis is one of the principal factors aiding in the evolution of the carcinogenic phenotype. A number of studies have implicated aberrant DNA methylation as a key survival mechanism in cancer, whereby promoter hypermethylation silences genes essential for many processes including apoptosis. To date, studies on the methylation profile of apoptotic genes have largely focused on cancers of the breast, colon and stomach, with only limited data available on prostate cancer. Here we discuss the major developments in the field of DNA methylation and its role in the regulation of aberrant apoptosis in prostate cancer. The most significant advances have involved the discovery of apoptotic gene targets of methylation, including XAF1, (fragile histidine triad (FHIT ), cellular retinol binding protein 1 (CRBP1), decoy receptor 1(DCR1), decoy receptor 2 (DCR2 ), target of methylation-induced silenceing 1 (TMS1), TNF receptor superfamily, member 6 (FAS), Reprimo (RPRM) and GLI pathogenesis-related 1 (GLIPR1). These genes are reported to be hypermethylated in prostate cancer and some offer potential as diagnostic and prognostic markers. We also introduce the concept of an 'apoptotic methylation signature' for prostate cancer and evaluate its potential in a diagnostic, prognostic and therapeutic setting.

FCHo proteins are nucleators of clathrin-mediated endocytosis

Clathrin-mediated endocytosis, the major pathway for ligand internalization into eukaryotic cells, is thought to be initiated by the clustering of clathrin and adaptors around receptors destined for internalization. However, here we report that the membrane-sculpting F-BAR domain-containing Fer/Cip4 homology domain-only proteins 1 and 2 (FCHo1/2) were required for plasma membrane clathrin-coated vesicle (CCV) budding and marked sites of CCV formation. Changes in FCHo1/2 expression levels correlated directly with numbers of CCV budding events, ligand endocytosis, and synaptic vesicle marker recycling. FCHo1/2 proteins bound specifically to the plasma membrane and recruited the scaffold proteins eps15 and intersectin, which in turn engaged the adaptor complex AP2. The FCHo F-BAR membrane-bending activity was required, leading to the proposal that FCHo1/2 sculpt the initial bud site and recruit the clathrin machinery for CCV formation.

Synapsin- and actin-dependent frequency enhancement in mouse hippocampal mossy fiber synapses

The synapsin proteins have different roles in excitatory and inhibitory synaptic terminals. We demonstrate a differential role between types of excitatory terminals. Structural and functional aspects of the hippocampal mossy fiber (MF) synapses were studied in wild-type (WT) mice and in synapsin double-knockout mice (DKO). A severe reduction in the number of synaptic vesicles situated more than 100 nm away from the presynaptic membrane active zone was found in the synapsin DKO animals. The ultrastructural level gave concomitant reduction in F-actin immunoreactivity observed at the periactive endocytic zone of the MF terminals. Frequency facilitation was normal in synapsin DKO mice at low firing rates (approximately 0.1 Hz) but was impaired at firing rates within the physiological range (approximately 2 Hz). Synapses made by associational/commissural fibers showed comparatively small frequency facilitation at the same frequencies. Synapsin-dependent facilitation in MF synapses of WT mice was attenuated by blocking F-actin polymerization with cytochalasin B in hippocampal slices. Synapsin III, selectively seen in MF synapses, is enriched specifically in the area adjacent to the synaptic cleft. This may underlie the ability of synapsin III to promote synaptic depression, contributing to the reduced frequency facilitation observed in the absence of synapsins I and II.

Differential efficiency of the endocytic machinery in tonic and phasic synapses

Efficient synaptic vesicle membrane recycling is one of the key factors required to sustain neurotransmission. We investigated potential differences in the compensatory endocytic machineries in two glutamatergic synapses with phasic and tonic patterns of activity in the lamprey spinal cord. Post-embedding immunocytochemistry demonstrated that proteins involved in synaptic vesicle recycling, including dynamin, intersectin, and synapsin, occur at higher levels (labeling per vesicle) in tonic dorsal column synapses than in phasic reticulospinal synapses. Synaptic vesicle protein 2 occurred at similar levels in the two types of synapse. After challenging the synapses with high potassium stimulation for 30 min the vesicle pool in the tonic synapse was maintained at a normal level, while that in the phasic synapse was partly depleted along with expansion of the plasma membrane and accumulation of clathrin-coated intermediates at the periactive zone. Thus, our results indicate that an increased efficiency of the endocytic machinery in a synapse may be one of the factors underlying the ability to sustain neurotransmission at high rates.

Navigating and exploring the ASD-friendly landscape

As a society we have a responsibility to provide an inclusive built environment. For those with Autistic Spectrum Disorder (ASD) however, the world can be a frightening, difficult and confusing place. The challenge of integrating more fully into society can be distanced by an alienating built environment. This is particularly debilitating for younger children who can find themselves detached from learning and interaction with their peers by uncomfortable surroundings. Subsequently there has been a growing interest in promoting ASD-friendly environments, especially in a school setting. Strategies to date have generally followed a widely accepted reductionist or generalist approach. However, the authors now contend that there needs to be a greater discussion of what truly constitutes an ASD-friendly environment, in conjunction with investigating what strategies best articulate a progressive approach to supporting those, and especially the young, with ASD in our built environment. Hence this paper first introduces some of the challenges faced by those with ASD in trying to cope with their surroundings. It then outlines a triad of challenges to overcome when considering what truly constitutes an ASD-friendly environment. The authors then highlight the need and advantage of supporting change and adaption in our shared inhabited landscape through providing both choice and reassurance for the child with ASD. It is hoped that by increasing awareness and then questioning what genuinely constitutes an ASD-friendly environment, it might ultimately help facilitate greater inclusion of the ASD child into mainstream education and society at large.

Sustained intraocular VEGF neutralization results in retinal neurodegeneration in the Ins2Akita diabetic mouse

Current therapies that target vascular endothelial growth factor (VEGF) have become a mainstream therapy for the management of diabetic macular oedema. The treatment involves monthly repeated intravitreal injections of VEGF inhibitors. VEGF is an important growth factor for many retinal cells, including different types of neurons. In this study, we investigated the adverse effect of multiple intravitreal anti-VEGF injections (200 ng/μl/eye anti-mouse VEGF164, once every 2 weeks totalling 5-6 injections) to retinal neurons in Ins2(Akita) diabetic mice. Funduscopic examination revealed the development of cotton wool spot-like lesions in anti-VEGF treated Ins2(Akita) mice after 5 injections. Histological investigation showed focal swellings of retinal nerve fibres with neurofilament disruption. Furthermore, anti-VEGF-treated Ins2(Akita) mice exhibited impaired electroretinographic responses, characterized by reduced scotopic a- and b-wave and oscillatory potentials. Immunofluorescent staining revealed impairment of photoreceptors, disruptions of synaptic structures and loss of amacrine and retinal ganglion cells in anti-VEGF treated Ins2(Akita) mice. Anti-VEGF-treated WT mice also presented mild amacrine and ganglion cell death, but no overt abnormalities in photoreceptors and synaptic structures. At the vascular level, exacerbated albumin leakage was observed in anti-VEGF injected diabetic mice. Our results suggest that sustained intraocular VEGF neutralization induces retinal neurodegeneration and vascular damage in the diabetic eye.

Identification of human neutrophil defensins (HNPs1-3) in biopsies of squamous cell carcinomas of the human tongue

It has previously been reported that the a-defensins, found in the granules of polymorphonuclear leukocytes (neutrophils/ PMNs), are cytolytic for human tumour cells in vitro. Objective: To identify and quantify the a- defensins, HNP-1, HNP-2 and HNP-3 in healthy and tumour tissue from patients with oral squamous cell carcinoma using HPLC, mass spectrometry and amino acid sequencing. Methods: All patients (n=5) were diagnosed with oral squamous cell carcinoma of the tongue.Biopsy tissue from the site of the tumour (n=5) and a non-affected region of the tongue (n=5) was snap frozen and subsequently stored at -70 ºC until analysed. Peptides were extracted from the 10 tissue biopsies using acidified ethanol. Peptide extracts were separated by reverse-phase HPLC . All tumour and control tissue samples were individually analysed under identical conditions with a flow rate of l ml/min, ambient column temperature and absorbance detection at 214 and 280 nm. Fractions (1ml) were collected automatically. HPLC fractions were analysed by MALDI-MS using a linear time-of-flight Voyager DE-mass spectrometer (PerSeptive Biosystems, UK). Using this system the detection limit was 10 fmol. Peptides with molecular masses corresponding to those reported for the a-defensins were deemed of interest and were further subject to complete structural analysis by automated Edman degradation using an Applied Biosystems 491 Procise microsequencer. Results: MALDI-MS revealed a triad of peptides of molecular masses 3442 Da, 3371 Da and 3486 Da in both healthy and tumour tissue. Full length sequence data were obtained for the three a-defensins, unequivocally identifying their presence in both tumour and healthy tissue. Analysis of the MALDI-MS and sequence data indicated that the a-defensins were overexpressed (up to 12 fold) in tumour tissue. Conclusion: This study demonstrates the feasibility of screening tumour tissue for novel peptides/proteins using HPLC and MALDI-MS.The role of a-defensins in oral squamous cell carcinoma of the tongue requires further investigation.

MRZ-99030 – a novel modulator of Aβ aggregation: II – reversal of Aβ oligomer-induced deficits in long-term potentiation (LTP) and cognitive performance in rats and mice.

β-amyloid1-42 (Aβ1-42) is a major endogenous pathogen underlying the aetiology of Alzheimer's disease (AD). Recent evidence indicates that soluble Aβ oligomers, rather than plaques, are the major cause of synaptic dysfunction and neurodegeneration. Small molecules that suppress Aβ aggregation, reduce oligomer stability or promote off-pathway non-toxic oligomerization represent a promising alternative strategy for neuroprotection in AD. MRZ-99030 was recently identified as a dipeptide that modulates Aβ1-42 aggregation by triggering a non-amyloidogenic aggregation pathway, thereby reducing the amount of intermediate toxic soluble oligomeric Aβ species. The present study evaluated the relevance of these promising results with MRZ-99030 under pathophysiological conditions i.e. against the synaptotoxic effects of Aβ oligomers on hippocampal long term potentiation (LTP) and two different memory tasks. Aβ1-42 interferes with the glutamatergic system and with neuronal Ca2+ signalling and abolishes the induction of LTP. Here we demonstrate that MRZ-99030 (100–500 nM) at a 10:1 stoichiometric excess to Aβ clearly reversed the synaptotoxic effects of Aβ1-42 oligomers on CA1-LTP in murine hippocampal slices. Co-application of MRZ-99030 also prevented the two-fold increase in resting Ca2+ levels in pyramidal neuron dendrites and spines triggered by Aβ1-42 oligomers. In anaesthetized rats, pre-administration of MRZ-99030 (50 mg/kg s.c.) protected against deficits in hippocampal LTP following i.c.v. injection of oligomeric Aβ1-42. Furthermore, similar treatment significantly ameliorated cognitive deficits in an object recognition task and under an alternating lever cyclic ratio schedule after the i.c.v. application of Aβ1-42 and 7PA2 conditioned medium, respectively. Altogether, these results demonstrate the potential therapeutic benefit of MRZ-99030 in AD.

A Burkholderia Type VI Effector Deamidates Rho GTPases to Activate the Pyrin Inflammasome

Burkholderia cenocepacia is an opportunistic pathogen of the cystic fibrosis lung that elicits a strong inflammatory response. B. cenocepacia employs a type VI secretion system (T6SS) to survive in macrophages by disarming Rho-type GTPases, causing actin cytoskeletal defects. Here, we identified TecA, a non-VgrG T6SS effector responsible for actin disruption. TecA and other bacterial homologs bear a cysteine protease-like catalytic triad, which inactivates Rho GTPases by deamidating a conserved asparagine in the GTPase switch-I region. RhoA deamidation induces caspase-1 inflammasome activation, which is mediated by the familial Mediterranean fever disease protein Pyrin. In mouse infection, the deamidase activity of TecA is necessary and sufficient for B. cenocepacia-triggered lung inflammation and also protects mice from lethal B. cenocepacia infection. Therefore, Burkholderia TecA is a T6SS effector that modifies a eukaryotic target through an asparagine deamidase activity, which in turn elicits host cell death and inflammation through activation of the Pyrin inflammasome.

A atitude diagnóstica enquanto instrumento de análise de ação educativa

A investigação realizada encontra-se inserida na área das Ciências de Educação, teve como objetivo principal compreender a atitude diagnóstica no quadro das situações educativas e pedagógicas desenvolvidas pelas educadoras de alguns jardins-de-infância, do distrito de Évora e como finalidade construir uma teoria de médio alcance elaborada numa estrutura explicativa do conjunto de fenómenos em contexto. Em educação de infância, o ato de cuidar estabelece-se numa relação de ajuda à criança e orienta-se para objetivos como promover o bem-estar, o conforto e o desenvolvimento em todas as dimensões. Isto implica conhecer e compreender cada criança ou grupo de crianças e as situações educativas e pedagógicas, em contexto. Compreende um exercício prático e para o qual se projeta uma prática de diagnóstico conciliadora da compreensão dos fenómenos sociais, estruturados em comportamentos, ações e atitudes específicas. Verificam-se lacunas quanto à forma como este exercício se cumpre e evolui nas diferentes fases. Repetem-se fragilidades quanto à mobilização de saberes, atitudes e competências a cada momento do agir. Constatam-se conceitos psicologizados e estruturas muito ténues e, por isso, necessitam ser estudadas, documentadas e teorizadas, particularmente numa dimensão praxiológica do conhecimento. O estudo inseriu-se no quadro das metodologias qualitativas, seguiu o paradigma interpretativo e o raciocínio indutivo. O referencial metodológico reuniu os princípios e procedimentos da Grounded Theory. A colheita de dados efetuou-se em jardins-de-infância da rede pública; a amostra teórica envolveu seis educadoras de infância e as crianças com idades compreendidas entre os três e os seis anos. Adotámos o uso de multitécnicas, entre as quais, a observação, a entrevista e a narrativa escrita. Fizemos vinte observações, repartidas pelos dois períodos do dia (manhã e tarde); vinte e seis entrevistas e seis narrativas escritas. Da análise dos dados emergiu a Atitude Diagnóstica como uma predisposição que caracteriza o ato de agir e as suas características foram-se tornando evidentes com o desenvolvimento da caracterização do processo de diagnóstico. Desta emergiu o modelo teórico definido em três eixos fundamentais. O “Processo de avaliação diagnóstica e planeamento” representativo das etapas, segundo as quais o educador desenvolve um conjunto de ações propiciadoras de um conhecimento previamente organizado, visando o bem-estar, o conforto, a segurança e o desenvolvimento. Em concomitância emerge o segundo, “Processo de intervenção educativa e pedagógica”, expressivo do conjunto de ações coerentes e evolutivas, empreendidas com vista à execução dos objetivos do ensino aprendizagem. Trata-se de dois processos sistematizados e perspetivados sob as dimensões diacrónica e sincrónica, compostos por uma sequência de pensamentos, permanentemente averbados pela “Atitude Diagnóstica” que dá um caráter coerente e evolutivo às tomadas de decisão nas ações educativas. Estes dois eixos integrados e entrelaçados são auxiliados por um terceiro, o “Processo de relação” que os harmoniza e dá especificidade a cada situação experienciada. A relação consolida-se e assume-se uma ajuda na confiabilidade necessária para a promoção da confiança entre os pares e o conhecimento vai evoluindo gradualmente em função do tempo e dos compromissos. O educador age com intencionalidade e para cada ator traça objetivos a cada momento do agir. A gestão do tempo, dos sentimentos e emoções funciona como variável importante na relação orientada sob a tríade: educador, criança e família e é concomitante com o Desenvolvimento pessoal e profissional do educador; ### ABSTRACT: The Diagnostic Attitude as an Analysis Instrument in Educational Action The conducted investigation is inserted in the area of Education Sciences. Its main aim was to understand the diagnostic attitude in the frame of the educative and pedagogical activities developed by the educators of some kindergartens in the district of Évora, and its purpose was to build a medium range theory elaborated in a structure explaining the whole of the phenomena in context. In child education, the act of caring is established in a relationship of help towards the child and is oriented towards goals as specific as promoting the well-being, the comfort and the development in all dimensions. This implies knowing and understanding each child or group of children and the educational and pedagogical situations in context. It includes a practical exercise that determines a complex scenario and for which we project a conciliating diagnostic practice towards the understanding of the social phenomena, structured in specific behaviors, actions and attitudes. We recognize some gaps regarding the way how this exercise is fulfilled and evolves in the different phases. Weaknesses regarding the mobilization of knowledge, attitudes and competences in each acting moment are repeated. We notice psichologized concepts and very superficial structures that need thus to be studied, documented and theorized, particularly in a praxeological dimension of knowledge. The study is inserted in the frame of qualitative methodologies, following an interpretative paradigm and an inductive reasoning. The methodological referential has gathered the principles and procedures of the Grounded Theory. The gathering of data was done in public kindergartens; the theoretical sample involved six child educators and children aged between three and six years. Several techniques were used, such as observation, interview and written narrative. We did twenty observations, divided between the two periods of the day (morning and afternoon); twenty-six interviews and six written narratives. The analysis of the data resulted in the Diagnostic attitude, like a predisposition that characterizes the act of acting and its characteristics became evident with the development of the characterization of the diagnostic process. From this rose the theoretical model defined in three fundamental axes. The Process of diagnostic evaluation and planning that represents the stages according to which the educator develops a group of actions that allow the transmission of a previously organized knowledge that aims well-being, comfort, safety and development. Concomitantly arises the second, Process of educative and pedagogic intervention, that expresses a group of coherent and evolving actions, undertaken to reach the goals of the teaching-learning process. These are two systematized processes, perspectivated under the diachronical and synchronical dimensions, composed by a sequence of thought, permanently confirmed by the Diagnostic attitude that gives a coherent and evolving character to the decision making in educative actions. These two integrated and intertwined axis are aided by a third one, Process of relation that harmonizes them and gives specificity to each experienced situation. The relation is consolidated and one assumes a kind of help in the trustworthiness necessary for the promotion of the trust between pairs and the knowledge keeps evolving gradually according to time and commitments. The educator acts with intent and sets goals for each actor at each acting moment. Time, feeling and emotion management works as an important variable in the relation oriented according to the triad educator, child and family, being also concomitant with the Personal and professional development of the educator.

Progesterone actions in protein phosphorylation in the central nervous system

O presente trabalho propõe-se esclarecer o papel que a progesterona e os seus metabolitos exercem no sistema nervoso central. Nos últimos anos, com a descoberta da síntese local de esteróides no cérebro, a progesterona, assim como outras hormonas sexuais, ganharam uma relevância crescente em fenómenos tais como plasticidade neuronal e neuroprotecção. Ainda que já se comece a entender o papel de muitas hormonas no cérebro, tal como o estrogénio, o papel da progesterona continua menos conhecido. Deste modo, o nosso trabalho centrou-se na elucidação dos efeitos da progesterona em fenómenos de sobrevivência celular, plasticidade neuronal/sináptica. Graças à colaboração com um grupo pioneiro em estudos sobre hormonas sexuais neuroactivas, o presente trabalho fornece uma importante contribuição ao entendimento do papel desta hormona no sistema nervoso central. Este trabalho fornece novos dados, relativamente ao papel da progesterona e dos seus metabolitos reduzidos na regulação de vias de sinalização associadas com sobrevivência celular, tal como Akt/PI3K e ERK. Também é analisado o efeito do tratamento hormonal na expressão e estado de fosforilação da proteína Tau, sendo ainda motivo de estudo cinases e fosfatases envolvidas nestes mecanismos.

Pancreatic zymogen granules: new proteins in unexpected places

Os mecanismos de biogénese, distribuição apical e secreção regulada de enzimas digestivas dos grânulos de zimogénio são, atualmente, pouco conhecidos. De modo a esclarecer e descrever estes processos de elevada importância biológica e clínica, é necessária uma melhor compreensão dos componentes da membrana granular e as funções e interações destes. Neste trabalho, através de uma abordagem proteómica, foi possível identificar novas proteínas granulares previamente associadas ao transporte vesicular sináptico. Para estudar as funções destas proteínas na génese e secreção de grânulos, foram realizados estudos de sobre-expressão, assim como estudos bioquímicos (1D, 2D, and LC-MS/MS) e morfológicos, utilizando céluas de mamífero. Entre as proteínas descobertas, cinco foram selecionadas e analisadas: RMCP-1, Piccolo, Synaptojanin-1, APP e ZG16p. Destas proteínas, confirmou-se a presença da RMCP-1 e APP nos grânulos de zimogénio. Interessantamente, o lectin ZG16p da secreção pâncreatico, encontra-se expressa no cérebro de rato, estando localizada nos terminais pós-sinápticos e em grânulos de RNA, indicando uma possível função desta proteína na formação das vesículas sinápticas. Finalmente, demonstrei que a formação de grânulos de zimogénio pode ser modulada, no modelo de células pancreáticas AR42J, pelas condições de cultura. Em contraste com as proteínas de carga neuroendocrinas, a sobreexpressão de proteínas de carga ou da membrana dos grânulos de zimogénio não foi suficiente para induzir a formação de grânulos ou de estruturas granulares em células constitutivamente secretoras, indicando diferenças na biogénese de grânulos neuroendócrinos e exócrinos.