A role for the small GTPases RAC1 and RAC1b in the modulation of NIS expression: potentiation of therapy with radioactive iodine in differentiated thyroid carcinoma

Introduction or Background: The Sodium Iodide Symporter (NIS) is responsible for active transport of iodide into thyroid follicular cells. The retention of its functional expression in most of the well-differentiated thyroid carcinomas (DTCs) enables the use of radioactive iodine (RAI) for treatment of metastatic disease. Still, about 30% of patients with advanced forms of DTC became refractory to RAI which makes their management very challenging. The main reason for impaired iodide uptake in refractory-DTC is the defective functional expression of NIS. Several molecular players have been described as critical for TSH-induced NIS expression, an example being the p38 mitogenic kinase. In breast cancer cells, the small GTPase RAC1 was shown to mediate the positive impact of p38 kinase activity on NIS expression. We, on the other hand, have previously shown that overexpression of RAC1b, a tumor-related splicing variant of RAC1, is associated with worse outcomes in DTC and correlates with the MAPK-activating BRAFV600E mutation, which has been related to the loss of NIS. Since RAC1 and RAC1b may act in an antagonistic fashion to regulate specific cellular responses, we asked if RAC1b would be implicated in NIS downregulation observed in DTCs. Methods Section: NIS expression levels were analyzed by RT-qPCR in a RAC1/RAC1b expression model system developed in non-transformed thyroid cell lines. A non-radioactive iodide influx assay was used to confirm the impact of RAC1-signaling on the efficacy of iodide uptake. Results Section: We demonstrate that ectopic overexpression of RAC1b is sufficient to decrease TSH-induced NIS expression, antagonizing the positive effect of RAC1 GTPase. Moreover, we clearly document, for the first time in thyroid cells, that both NIS expression and iodide uptake are downregulated upon RAC1 inhibition, supporting the role of canonical RAC1 signaling in promoting TSH-induced NIS expression. Conclusion: Our findings provide evidence that RAC1 and RAC1b signaling are implicated in the regulation of NIS expression in thyroid cells and suggest that RAC1b overexpression may be one of the mechanisms contributing to the low levels of NIS observed in some subgroups of DTCs, antagonizing RAC1 stimulatory effect on the TSH/cAMP-mediated induction of NIS expression.FCT - PTDC/BIAMOL/31787/2017N/

Longitudinal effects of ketamine on cell proliferation and death in the CNS of zebrafish

Funding Information: This work was funded by FEDER funds through the Operational Competitiveness Programme – COMPETE and by National Funds through FCT – Portuguese Foundation for Science and Technology under the project FCOMP-01-0124-FEDER-028683 (PTDC/CVT-WEL/4672/2012). Publisher Copyright: © 2023 The AuthorsZebrafish is known for its widespread neurogenesis and regenerative capacity, as well as several biological advantages, which turned it into a relevant animal model in several areas of research, namely in toxicological studies. Ketamine is a well-known anesthetic used both in human as well as veterinary medicine, due to its safety, short duration and unique mode of action. However, ketamine administration is associated with neurotoxic effects and neuronal death, which renders its use on pediatric medicine problematic. Thus, the evaluation of ketamine effects administration at early stages of neurogenesis is of pivotal importance. The 1‐41–4 somites stage of zebrafish embryo development corresponds to the beginning of segmentation and formation of neural tube. In this species, as well as in other vertebrates, longitudinal studies are scarce, and the evaluation of ketamine long-term effects in adults is poorly understood. This study aimed to assess the effects of ketamine administration at the 1–4 somites stage, both in subanesthetic and anesthetic concentrations, in brain cellular proliferation, pluripotency and death mechanisms in place during early and adult neurogenesis. For that purpose, embryos at the 1–4 somites stage (10.5 h post fertilization - hpf) were distributed into study groups and exposed for 20 min to ketamine concentrations at 0.2/0.8 mg/mL. Animals were grown until defined check points, namely 50 hpf, 144 hpf and 7 months adults. The assessment of the expression and distribution patterns of proliferating cell nuclear antigen (PCNA), of sex-determining region Y-box 2 (Sox 2), apoptosis-inducing factor (AIF) and microtubule-associated protein 1 light chain 3 (LC3) was performed by Western-blot and immunohistochemistry. The results evidenced the main alterations in 144 hpf larvae, namely in autophagy and in cellular proliferation at the highest concentration of ketamine (0.8 mg/mL). Nonetheless, in adults no significant alterations were seen, pointing to a return to a homeostatic stage. This study allowed clarifying some of the aspects pertaining the longitudinal effects of ketamine administration regarding the CNS capacity to proliferate and activate the appropriate cell death and repair mechanisms leading to homeostasis in zebrafish. Moreover, the results indicate that ketamine administration at 1–4 somites stage in the subanesthetic and anesthetic concentrations despite some transitory detrimental effects at 144 hpf, is long-term safe for CNS, which are newly and promising results in this research field.publishersversionpublishe

Pseudotumor Pancreático – Um caso de Tuberculose Abdominal

Tuberculosis is a frequent disease caused by Mycobacterium tuberculosis and can be present in every organ. (1,2) The authors report a 75 years old woman case of abdominal tuberculosis, who had clinical e imagiological suspicion of pancreatic cancer and in whom surgery was the key for diagnosis and treatment. Keywords: Pancreatic tuberculosis, Peripancreatic tuberculosis, Pancreatic tumour A tuberculose é uma doença causada por agentes pertencentes ao complexo constituído pelo Mycobacterium tuberculosis, que têm uma distribuição universal no organismo, podendo atingir qualquer órgão. (1,2) Os autores apresentam um caso clínico de tuberculose abdominal numa doente de 75 anos de idade, sem antecedentes de risco, na qual os exames complementares de diagnóstico orientavam para o diagnóstico de neoplasia pancreática. A cirurgia, que consistiu em biópsia excisional da lesão, foi decisiva para o diagnóstico e para o tratamento da doente. Palavras-chave: Tuberculose pancreática, Tuberculose Peripancreática, Tumor do Pâncreas.

[Carotid atherosclerosis and white matter hypodensities: a controversial relationship]

INTRODUCTION: White matter hypodensities of presumed vascular origin, are recognized as an important cause of morbidity with established clinical and cognitive consequences. Nonetheless, many doubts remain on its physiopathology. Our goal is to clarify the potential role of carotid atherosclerosis and other vascular risk factors in the development of white matter hypodensities of presumed vascular origin. MATERIAL AND METHODS: We included patients that underwent CT brain scan and neurosonologic evaluation within a one-month period. Full assessment of vascular risks factors was performed. We seek to find independent associations between white matter hypodensities of presumed vascular origin, carotid intima-media thickness and vascular risk factors. RESULTS: 472 patients were included, mean age was 67.32 (SD: 14.75), 274 (58.1%) were male. The independent predictors of white matter hypodensities of presumed vascular origin were age (OR: 1.067, 95% IC: 1.049 - 1.086, p < 0.001) and hypertension (OR: 1.726, 95% IC: 1.097 - 2.715, p = 0.018). No association was found between IMT (OR: 2.613, 95% IC: 0.886 - 7.708, p = 0.082) or carotid artery stenosis (OR: 1.021, 95% IC: 0.785 - 1.328, p = 0.877) and white matter hypodensities of presumed vascular origin. DISCUSSION: Only age and hypertension proved to have an independent association with white matter hypodensities of presumed vascular origin. Carotid atherosclerosis, evaluated by IMT and the degree of carotid artery stenosis, showed no association with white matter hypodensities of presumed vascular origin. Since atherosclerosis is a systemic pathology, these results suggest that alternative mechanisms are responsible for the development of white matter hypodensities of presumed vascular origin. CONCLUSION: Age and hypertension seem to be the main factors in the development of white matter hypodensities of presumed vascular origin. No association was found between carotid atherosclerosis and white matter hypodensities of presumed vascular origin

Modelling the habitat preferences of the NE-Atlantic Sea cucumber Holothuria forskali : demographics and abundance

This work was funded by the Operational Program Mar2020 MAR-02.01.01-FEAMP-0052 “Newcumber - Avanços para o cultivo sustentável de pepinos do mar”. This work was also supported by national funds through FCT - Fundação para a Ciência e a Tecnologia, I.P., within the scope of the projects (https://doi.org/10.54499/UIDB/04292/2020, and https://doi.org/10.54499/UIDB/00006/2020) and the Associate Laboratory ARNET (https://doi.org/10.54499/LA/P/0069/2020), A.C. Brito with the Scientific Stimulus Program – CEECIND/00095/2017, A.P. with the under the Scientific Employment Stimulus - Institutional Call - CEECINST/00051/2018 and Francisco Azevedo e Silva and João Trigo de Sousa through the individual grants (SFRH/BD/09563/2020 and SFRH/BDANA/02949/2023).Sea cucumbers' historical demand, together with the depletion of several traditional species in the market, has popularized new target species from new fishing grounds. Holothuria forskali is one of those emergent species in the trade market. However, it is a species for which there is no relevant information to allow sustainable stock management. Fundamental knowledge of the populations' structure and habitat preferences are key elements without which any measure is inconsequent. This work aims to fill that gap by modelling temporal and spatial patterns of abundance and demographic structure of this species in a NE-Atlantic area, as a function of environmental features. For a period of 15 months, nine regular sampling campaigns collected data on density, individual length, individual conditions of occurrence (e.g. sheltered, on sand, on algae cover) and environmental parameters (water column, sediment, substrate cover and type), using random transects throughout a costal rocky-reef, considering habitat heterogeneity and substrate types. To determine the species' habitat preferences Generalized Linear Models were used to model density and demographic structure of the species as a function of environmental conditions. The models revealed that the main drivers shaping the distribution of H. forskali are neither abiotic nor biotic parameters of the water column, but physical stressors, like current intensity and depth, and substrate type in a patchy distribution pattern. Estuarine conditions are generally avoided, although with a size-dependent opportunistic strategy. Larger individuals show temporal and spatial displacement patterns towards suitable reproductive conditions (pre-breeding aggregation) and favourable feeding grounds and smaller size-classes tend to aggregate in higher numbers in more stable environments. Sustainable sources for market supply, like aquaculture, are still a long way from commercial production. So, these results are fundamental to support effective conservation measures for stock management of H. forskaliPublisher PDFPeer reviewe

Allometric relationships to assess ontogenetic adaptative changes in three NE Atlantic commercial sea cucumbers (Echinodermata, Holothuroidea)

This study was financed by the Operational Program Mar2020 nº MAR-02.01.01-FEAMP-0052 “Newcumber—Avanços para o cultivo sustentável de pepinos-do-mar”. It received further financial support from Fundação para a Ciência e a Tecnologia with project UIDB/04292/2020; A.C. Brito and A. Pombo through the Scientific Employment Stimulus Programmes (CEECIND/00095/2017 and CEECINST/00051/2018); and Francisco Azevedo e Silva through the individual research Grant 2020.09563.BD; Also, T. A. Marques and C. Rocha thank partial support by CEAUL (funded by FCT—Fundação para a Ciência e a Tecnologia, Portugal, through the project UIDB/00006/2020).Holothuria arguinensis, Holothuria mammata and Holothuria forskali are three common sea cucumber species found in the NE Atlantic, traded in international markets and susceptible to capture. Allometric relationships reveal if the scaling relationships between biometric characters are proportional with growth, being a useful tool to understand species growth strategies. Allometric relationships of the three species were estimated and compared between them and with populations from different regions. These allometric relationships revealed that the three species have negative allometric growth. However, they have different growth strategies between them and reveal different regional intra-specific growth strategies when compared to other populations, suggesting ontogenetic adaptation as a consequence of external factors. PostprintPeer reviewe

La evaluación de la calidad de enseñanza universitaria mediante la percepción del alumno de una actividad práctica: propuesta de escalas de medida

El panorama universitario actual, centrado en la valoración y juicio de los distintos agentes del proceso de enseñanza-aprendizaje, supone un campo adecuado para el desarrollo de investigaciones que pretendan alcanzar indicadores fiables del desempeño de las labores docentes. La futura implantación del Espacio Europeo de Educación Superior (EEES), acrecienta el interés por este tipo de investigaciones.Este trabajo, centrado en este contexto, pretende evaluar y valorar, por parte del discente, una actividad práctica constitutiva del programa de dos asignaturas universitarias impartidas en dos licenciaturas oficiales. Para su evaluación desarrollaremos una escala de medida que permita conocer la percepción de calidad de dicha actividad por parte de los alumnos y las posibles fuentes de mejora. Con los resultados obtenidos podremos conocer que aportación tiene esta práctica en el proceso de aprendizaje de esta materia y para la formación general del alumno

Twenty years of a pre-symptomatic testing protocol for late-onset neurological diseases in Portugal

Introduction: The national protocol of genetic counselling and pre-symptomatic testing for late-onset neurological diseases began in Portugal in 1995. Initially, it was accessible only to adults at-risk for Machado-Joseph disease, but was later extended to other hereditary ataxias, to Huntington’s disease and to familial amyloid polyneuropathy caused by Val30Met mutation at the transthyretin gene. The aim of this study was to describe the profile of the population seeking pre-symptomatic testing, while also reflecting on the experience of conducting the protocol of multidisciplinary sessions since 1996. Material and Methods: We conducted a retrospective study and collected data from clinical records of consultands who requested pre-symptomatic testing at our centre in Porto (Portugal) during the first twenty years of practice (1996 - 2015). Results: A total of 1446 records were reviewed. The most common reason for testing was to reduce uncertainty (41.7%). The rate of withdrawals before results disclosure was lower (16%) than reported in other international experiences with pre-symptomatic testing, while 45% of the consultands dropped out the protocol after learning the test results (73.5% of them were non-carriers). As far as the mutation carriers were concerned, 29.6% adhered to the protocol a year after test disclosure. Consultands that had learned about presymptomatic testing through healthcare professionals tended to adhere more to pre-symptomatic testing consultations. Discussion: The profile of Portuguese consultands at risk for late-onset neurological diseases is similar to those reported in other international programs. The largest group in this data set was the one comprising the subjects at risk for familial amyloid polyneuropathy caused by Val30Met mutation at the transthyretin gene, and it is likely that therapeutic options for this condition may have influenced this result. Adherence to pre-symptomatic testing may change in the future since effective therapies are available (or given the fact that people think effective treatments are imminent). Conclusion: This study reflects the first comprehensive description of a Portuguese experience with pre-symptomatic testing for lateonset neurological diseases. The development of innovative approaches to improve the consultands’ experience with pre-symptomatic testing and their engagement in genetic departments is still a challenge in Portuguese genetics healthcare departments. A better coordination among primary care and genetics healthcare services is needed