Explore the vast range of titles available.

Natural killer (NK) cells are innate lymphoid cells involved in tumor surveillance. These immune cells have the potential to fight cancer growth and metastasis, as such, their deregulation can result in tumor immune escape. Recently exosomes were described as mediators of intercellular communication between cancer and NK cells. The exact role of this subclass of extracellular vesicles (EVs), which transport genetic and molecular material to recipient cells, in NK cell biology in the context of cancer, is still an open question. Several reports have demonstrated that tumor-derived exosomes (TDEs) can exert immunomodulatory activities, including immunosuppression, thus promoting cancer progression. Some reports demonstrate that the interplay between cancer exosomes and NK cells allows tumors to escape immune regulation. On the other hand, tumor exosomes were also described to activate NK cells. Additionally, studies show that NK cell exosomes can modulate the immune system, opening up their potential as an immunotherapeutic strategy for cancer treatment. Our review will focus on the reprogramming effect of cancer exosomes on NK cells, and the immunotherapeutic potential of NK cells-derived exosomes.

The urgent need for rapid antimicrobial susceptibility is broadly apparent from government reports to the lay press. Accordingly, we developed a flow-cytometry assay (FCM) for evaluating ceftolozane-tazobactam (C/T) susceptibility directly on blood cultures (BC) requiring < 2 h from flag positivity to report. The protocol was optimized with C/T-susceptible and C/T-resistant gram-negative bacilli inoculated in BC aerobic bottles (Becton-Dickinson, USA), and afterward optimized for different C/T concentrations (1/4, 2/4, 4/4, and 8/4 mg/L) for 1 h incubation (37 °C), followed by FCM and software analysis. Fluorescent membrane permeability and membrane potential dyes were comparatively used to detect early cell lesions using the CytoFLEX cytometer (Beckman-Coulter, USA). Repeatability, reproducibility, and stability of the assay up to 48 h after BC positivity were determined. Internal validation was performed in spiked BC bottles with 130 Enterobacterales and 32 Pseudomonas aeruginosa isolates from Porto University (Portugal), including 13 ATCC isolates. Additionally, 64 gram-negative bacilli recovered from positive BC at Ramon y Cajal Hospital (Madrid, Spain) were tested. Categorical agreement (CA) and analytical errors were calculated comparing FCM with broth microdilution results. Only the membrane potential dyes clearly distinguished CT-susceptible and CT-resistant isolates. Excellent repeatability, reproducibility, and inter-method concordance was observed. Overall, CA was 99.1% using EUCAST criteria with 2 major errors and 98.7% with CLSI criteria with 2 major and 1 minor errors. A new, accurate, and ultra-rapid FCM (< 2 h) for testing C/T susceptibility gave accurate results and would expand current FCM antimicrobial susceptibility assay.

Trafficking protein particle (TRAPP) complexes, which include the TRAPPC4 protein, regulate membrane trafficking between lipid organelles in a process termed vesicular tethering. TRAPPC4 was recently implicated in a recessive neurodevelopmental condition in four unrelated families due to a shared c.454+3A>G splice variant. Here, we report 23 patients from 17 independent families with an early-infantile-onset neurodegenerative presentation, where we also identified the homozygous variant hg38:11:119020256 A>G (NM_016146.5:c.454+3A>G) in TRAPPC4 through exome or genome sequencing. No other clinically relevant TRAPPC4 variants were identified among any of over 10,000 patients with neurodevelopmental conditions. We found the carrier frequency of TRAPPC4 c.454+3A>G was 2.4–5.4 per 10,000 healthy individuals. Affected individuals with the homozygous TRAPPC4 c.454+3A>G variant showed profound psychomotor delay, developmental regression, early-onset epilepsy, microcephaly and progressive spastic tetraplegia. Based upon RNA sequencing, the variant resulted in partial exon 3 skipping and generation of an aberrant transcript owing to use of a downstream cryptic splice donor site, predicting a premature stop codon and nonsense mediated decay. These data confirm the pathogenicity of the TRAPPC4 c.454+3A>G variant, and refine the clinical presentation of TRAPPC4-related encephalopathy.

Pathogenic variants in the NGLY1 gene are associated with a Congenital Disorder of Deglycosylation (CDDG) characterized by delays in reaching developmental milestones, complex hyperkinetic movement disorder, transient elevation of transaminases, and alacrima or hypolacrima. To date, only few cases of NGLY1 deficiency have been identified and reported in the literature. This report highlights a first child of non‐consanguineous parents with no relevant family history who presented with hypotonia and poor weight gain since birth. At 2 months, the child developed paroxysmal cervical dystonia, posteriorly resolving spontaneously by age of 3. Subsequently, delays in reaching developmental milestones, ataxia, dyskinesia, visual impairment due to cone rod retinal dystrophy, low triglycerides, and persistently elevated liver transaminases were observed. Extensive etiological investigation was performed, including array‐CGH and metabolic evaluation with no abnormalities to note. Trio whole exome analysis identified a homozygous pathogenic variant of the NGLY1 gene, c.1891del (p.Gln631Serfs*7), consistent with CDDG. Both parents were confirmed to be heterozygous carriers. The authors discuss in this case, the clinical presentation, the diagnostic challenges, and review other relevant NGLY1 deficiency cases previously reported in the literature. This case, along with the previous reported in the literature, indicates that pathogenic variants in NGLY1 cause a recognizable phenotype and should be considered in patients with a typical presentation. It also suggests that decreased sweating is not present universally in these patients.

We present the case of a boy in his middle childhood with gait ataxia and loss of reflexes with a 1-year onset. He had a background of an autism spectrum disorder but was otherwise healthy. A paediatric cardiology assessment was requested to investigate possible cardiac involvement associated to his neurological symptoms. Even though he had no cardiac symptoms and a normal electrocardiography, the echocardiogram revealed severe asymmetric left ventricle hypertrophy consistent with hypertrophic cardiomyopathy. This prompted genetic testing and the diagnosis of Friedreich’s ataxia was confirmed.

No presente relatório apresento o estágio que realizei com a empresa Auditaccount para concluir o Mestrado em Tradução e Interpretação Especializadas. Assim, o objetivo desde estágio foi desenvolver todos os conhecimentos adquiridos ao longo do meu percurso académico no ISCAP, melhorando as minhas competências especificamente na área de tradução e interpretação.Como tal, exponho o meu trabalho desenvolvido no âmbito deste estágio, intercalando a teoria com a prática da tradução e interpretação que tive a oportunidade de experienciar, particularmente no domínio jurídico. Deste modo, demonstro as diferenças entre diferentes sistemas e textos jurídicos, a problemática da equivalência e os diferentes desafios do tradutor e do intérprete.

The implementation and potential of ketogenic dietary therapies (KDTs) have changed over time. The organization of KDT services, the availability of multidisciplinary teams, resources and support for patients and families still vary widely around the world. This diversity is reflected by a lack of consistency in reported outcomes, optimization of using KDT and KDT compliance. To highlight the unmet needs for KDT services, the ERN EpiCARE Ketogenic Dietary Therapy Special Interest Group (KDT SIG) conducted an online survey on KDT implementation and utilization, addressing the following topics: Use and completeness of guidelines and protocols; assessment of compliance and outcome parameters, sustainability and inclusivity in daily life. Consistently reported unmet needs included the lack of psychological support and resources to measure and improve adherence to KDT, the lack of inclusion strategies, and shared guidelines and protocols adapting to specific needs. Future interventions should focus primarily on educational and informative measures together with creation of shared protocols for complex care. Plain Language Summary This study provides the results of a survey compiled by clinicians and patients representatives belonging to ERN Epicare, designed to unravel unmet needs from both patients' and healthcare practitioners' perspectives during ketogenic dietary therapies (KDT) provision. Importantly, results show the need to create new shared protocols and guidelines meant for KDT use in complex care situations and to develop future strategies initiatives to support patients improving their social inclusivity.

Exosomes are extracellular vesicles produced by all cell types in the endosomal compartment and are key players in intercellular communication. Exosomes are involved in tumorigenesis, metastasis formation, angiogenesis, disease progression, drug resistance and tumor immune escape. The exosomal content contributes for the reprograming of recipient cells, namely the immune cells, which confers exosomes an immunomodulatory potential. The establishment and development of PDAC is marked by a prominent recruitment of immunosuppressive cells and the lack of cytotoxic immune cells in the tumor microenvironment. However, how PDAC exosomes modulate the immune system and how the immune system influences PDAC establishment and progression are still fields yet to explore. To address these questions, we used genetically engineered mouse models (GEMMs) of pancreatic cancer: the KPC mouse model, which develops PDAC in a spontaneous manner, recapitulating the human disease; the KPC Rag2 GEMM, which is an immunodeficient mouse model lacking B and T cells in order to study the effect of these cells in PDAC establishment and progression; and the KPC Rag2 IL2rg mouse models, lacking B, T and NK cells, in an attempt to understand the role of NK cells in PDAC establishment and development. Moreover, in order to study the role of PDAC exosomes in immunomodulation, we have impaired exosomes secretion in a PDAC mouse model and analysed changes in the immune landscape of the tumours.The observations collected in this thesis suggest that the immune system is not blind to PDAC, and have uncovered preliminary data that indicates that cancer exosomes are involved in the anti-tumour immune response, opening new avenues for the use of immunotherapy in PDAC patients.

Introdução: A amiloidose hereditária por transtirretina associada à mutação V30M é uma doença autossómica dominante rara causada por uma mutação no gene da transtirretina (TTR). A mutação causa uma proteína anormal que sofre um misfolding e agregação, levando a uma deposição de fibrilhas insolúveis nos órgãos e nos tecidos, especialmente no sistema nervoso periférico, coração, rim e olho. Em consequência da deposição e acumulação de TTR, surgem as manifestações clínicas, que incluem a neuropatia periférica sensitivo-motora, neuropatia autonómica, cardiopatia, nefropatia, alterações oculares, entre outras. Uma vez que a TTR plasmática é quase exclusivamente o produto da síntese hepática, o transplante hepático tem sido utilizado como tratamento para a amiloidose por TTR para prevenir a síntese da TTR mutante plasmática. Porém, como a TTR também é produzida na retina e no corpo ciliar, as fontes oculares da TTR mutante não são interrompidas pelo transplante hepático, pelo que as manifestações oculares podem continuar a manifestar-se. A angiopatia amiloidótica retiniana (RAA) tende a surgir tardiamente no curso da doença. No que concerne ao edema macular, o uso de anti-VEGF é útil na medida em que estes agentes diminuem o edema. Porém, estão também associados a diversos efeitos secundários induzidos ou potenciados pelo VEGF. As injeções frequentes e a longo-prazo que são comummente necessárias podem aumentar o burdenpara os doentes e família e o risco de complicações oculares e sistémicas. Assim, tratamentos alternativos para o edema macular com maior conforto e segurança poderão ser vantajosos. O implante de FAc é um implante intravítreo não biodegradável concebido para uma duração de até 36 meses. O implante de FAc tem sido usado para o tratamento de doenças vasculares inflamatórias, nomeadamente o edema macular diabético, uveíte, edema macular cistóide na síndrome de Irvine-Gass e na oclusão da veia da retina.Objetivo: Com este trabalho, pretendeu-se avaliar a eficácia e a segurança do implante de FAc (0,2 µg/dia FAc, ILUVIEN®) no edema macular por RAA em doentes com amiloidose por ATTRv associada à mutação V30M.Métodos: Estudo retrospetivo observacional de uma série de casos realizado no Centro Hospitalar e Universitário de Santo António, Portugal. O estudo incluiu 15 doentes com o diagnóstico genético de amiloidose por ATTRv associada à mutação V30M (15 olhos), tratados para o edema macular com o implante intravítreo de FAc de 0,19 mg, entre janeiro de 2010 e dezembro de 2019. Os outcomes foram a melhor acuidade visual corrigida (BCVA), espessura central da fóvea (CFT) e pressão intraocular (PIO). Estes dados foram recolhidos no início do estudo (baseline), na semana 1 e nos meses 1, 3, 6 e 12.Resultados: Houve uma melhoria estatisticamente significativa na BCVA e na CFT em todos os momentos (semana 1, mês 3, 6, e 12) em relação à baseline. Adicionalmente, não houve nenhuma diferença estatisticamente significativa da IOP durante o período de follow-up, comparativamente à baseline.Conclusão: Os doentes com edema macular por RAA associada à mutação V30M tratados com o implante de FAc mostraram um aumento sustentado dos resultados funcionais e uma melhoria anatómica, comparativamente à baseline. A IOP manteve-se estável durante os 12 meses de follow-up.

Objective Clinical care of rare and complex epilepsies is challenging, because evidence‐based treatment guidelines are scarce, the experience of many physicians is limited, and interdisciplinary treatment of comorbidities is required. The pathomechanisms of rare epilepsies are, however, increasingly understood, which potentially fosters novel targeted therapies. The objectives of our survey were to obtain an overview of the clinical practice in European tertiary epilepsy centers treating patients with 5 arbitrarily selected rare epilepsies and to get an estimate of potentially available patients for future studies. Methods Members of the European Reference Network for rare and complex epilepsies (EpiCARE) were invited to participate in a web‐based survey on clinical practice of patients with Dravet syndrome, tuberous sclerosis complex (TSC), autoimmune encephalitis, and progressive myoclonic epilepsies including Unverricht Lundborg and Unverricht‐like diseases. A consensus‐based questionnaire was generated for each disease. Results Twenty‐six of 30 invited epilepsy centers participated. Cohorts were present in most responding centers for TSC (87%), Dravet syndrome (85%), and autoimmune encephalitis (71%). Patients with TSC and Dravet syndrome represented the largest cohorts in these centers. The antiseizure drug treatments were rather consistent across the centers especially with regard to Dravet syndrome, infantile spasms in TSC, and Unverricht Lundborg / Unverricht‐like disease. Available, widely used targeted therapies included everolimus in TSC and immunosuppressive therapies in autoimmune encephalitis. Screening for comorbidities was routinely done, but specific treatment protocols were lacking in most centers. Significance The survey summarizes the current clinical practice for selected rare epilepsies in tertiary European epilepsy centers and demonstrates consistency as well as heterogeneity in the treatment, underscoring the need for controlled trials and recommendations. The survey also provides estimates for potential participants of clinical trials recruited via EpiCARE, emphasizing the great potential of Reference Networks for future studies to evaluate new targeted therapies and to identify novel biomarkers.