The influence of physical exercise on oestrogen and androgen receptor expression in a chemically and hormonally-induced rat model of prostate cancer

Background: Oestrogen (ER) and androgen (AR) recep- tors play an important role in normal prostate development and are also implied in prostate cancer (PCa) development. Several studies suggested that physical activity may decrease the risk of PCa development and also changes sexual hor- mones and their receptors. This study aimed to evaluate the effects of physical exercise on ERα and AR expression in a rat model of chemically and hormonally-induced PCa. Materials and Methods: Fifty-five male Wistar Unilever rats of 12 weeks of age were randomly divided into four groups: control sedentary (n = 10), control exercised (n = 10), induced sedentary (n = 15) and induced exercised (n = 20). Animals from exercised groups started the exercise training in a treadmill (Treadmill Control LE 8710, Harvard Apparatus, USA), at the age of 8 weeks, for 35 weeks (5 days/week). The protocol for PCa induction started at 12 weeks of age and consisted of sequential administration of flutamide (50 mg/kg, TCI Chemicals), testosterone propion- ate (100 mg/kg, TCI Chemicals) and N-methyl-N-nitrosourea (30 mg/kg, Isopac®, Sigma Chemical Co.), followed by sub- cutaneous implants of crystalline testosterone. Animals were sacrificed at 61 weeks of age and a complete necropsy was performed. All experiments were approved by DGAV (no. 021326). Antibodies for Erα (1:500, clone 6F11, Novocastra) and AR (clone PG21, Merck Millipore) were used for the immunohistochemical study. The staining extension was evaluated in normal prostate tissue and in dorsolateral pros- tate lesions (hyperplasia, dysplasia, prostatic intraepithelial neoplasia (PIN) and microinvasive carcinoma) and assessed to five levels (0%, 75%), con- sidering the extension of immunopositive tissue. Data was analysed with SPSS 25.Results: The normal prostate tissue and dorsolateral prostate lesions of animals from all groups were immunopositive for Erα and AR. However, the groups showed high immunoposi- tivity for AR and low positivity for Erα ( 0.05). The malignant lesions (PIN and microinvasive carcinoma) showed lower AR expression when compared with normal prostate tissue in all groups. Conclusions: As expected, the AR expression was lower in malignant lesions. Inversely to that reported in other studies, the exercise training did not modify the ERα and AR expres- sion, which may be related to the duration and type of exer- cise performed

The novel ECF56 SigG1-RsfG system modulates morphological differentiation and metal-ion homeostasis in Streptomyces tsukubaensis

Extracytoplasmic function (ECF) sigma factors are key transcriptional regulators that prokaryotes have evolved to respond to environmental challenges. Streptomyces tsukubaensis harbours 42 ECFs to reprogram stress-responsive gene expression. Among them, SigG1 features a minimal conserved ECF s2–s4 architecture and an additional C-terminal extension that encodes a SnoaL_2 domain, which is characteristic for ECF s factors of group ECF56. Although proteins with such domain organisation are widely found among Actinobacteria, the functional role of ECFs with a fused SnoaL_2 domain remains unknown. Our results show that in addition to predicted self-regulatory intramolecular amino acid interactions between the SnoaL_2 domain and the ECF core, SigG1 activity is controlled by the cognate anti-sigma protein RsfG, encoded by a co-transcribed sigG1-neighbouring gene. Characterisation of ¿sigG1 and ¿rsfG strains combined with RNA-seq and ChIP-seq experiments, suggests the involvement of SigG1 in the morphological differentiation programme of S. tsukubaensis. SigG1 regulates the expression of alanine dehydrogenase, ald and the WhiB-like regulator, wblC required for differentiation, in addition to iron and copper trafficking systems. Overall, our work establishes a model in which the activity of a s factor of group ECF56, regulates morphogenesis and metal-ions homeostasis during development to ensure the timely progression of multicellular differentiation.This work was partially funded by National Funds through FCT-Fundação para a Ciência e a Tecnologia, I.P., under the project ERA-IB-2/0001/2015. It was further supported by FEDER - Fundo Europeu de Desen-volvimento Regional funds through the COMPETE 2020 - Operational Programme for Competitiveness and Internationalisation (POCI), Portugal 2020; and by Portuguese funds through FCT Fundação para a Ciência e a Tecnologia, I.P/Ministério da Ciência, Tecnologia e Ensino Superior POCI-01-0145-FEDER-007274 and NORTE-01-0145-FEDER-000012. BBSRC supported this work through the Institute Strategic Programme grant BB/J004561/1 to the John Innes Centre. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. R.O. was supported by the FCT fellowship SFRH/ BD/107862/2015 and by the EMBO fellowship ASTF438-2015, M.V.M. was supported by the FCT fellowship SFRH/BPD/95683/2013 and the FCT contract DL57/2016/CP1355/CT0023 and D.C.P. and G.F. were supported through the IMPRS-Mic and the ERASynBio project ECFexpress (BMBF grant 031L0010B). The authors are grateful to Kim Findlay at the Bioimaging platform of the John Innes Centre (JIC, UK) for performing the SEM imaging of S. tsukubaensis samples, Mervyn Bibb (JIC, UK) for the pIJ12333 plasmid , Mark Buttner (JIC, UK) for his comments and discussion regarding the work and Paula Tamagnini (i3S, PT) for comments on the manuscript. The authors acknowledge the support of the i3S Scientific Platforms Cell Culture and Genotyping, Biochemical and Biophysical Technologies and Proteomics

Effects of physical exercise in biochemical parameters and dorsolateral prostate lesions: data from a rat model of prostate cancer

Background: Prostate cancer (PCa) is among the most prevalent cancers worldwide. Physical exercise is widely recognized due to its beneficial effects. This study aimed to evaluate the effects of physical exercise on biochemical pa- rameters and in dorsolateral prostate lesions in a rat model of PCa. Materials and Methods: Ninety-five male Wistar Unilever rats were randomly divided into eight groups sacrificed at 35 (groups I) or 61 weeks of age (groups II): control sedentary groups (Cont+Sed I (n = 10); Cont+Sed II (n = 10)); induced sedentary group (PCa+Sed I (n = 10); PCa+Sed II (n = 15)); control exercised groups (Cont+EX I (n = 10); Cont+EX II (n = 10)) and induced exercised groups (PCa+EX I (n = 10); PCa+EX II (n = 20)). All procedures were approved (DGAV, no. 021326). Animals from exercised groups started the exer- cise program in a treadmill at 8 weeks of age, for 28 weeks or 53 weeks. The animals were trained 5 days/week, 60 min per day. Prostate lesions were induced at 12 weeks of age, with sequential administration of flutamide, testosterone propion- ate and N-methyl-N-nitrosourea, and subcutaneous implants of crystalline testosterone. Animals were sacrificed at 35 or 61 weeks of age. Peripheral blood of all animals was col- lected by intracardiac puncture. A complete necropsy was performed. The dorsolateral prostate tissues sections were processed for histological analysis. Data were analysed using SPSS 25. p 0.05). Dorsolateral prostate lesions were classified as dysplasia, prostatic intraep- ithelial neoplasia (PIN) and microinvasive carcinoma. The number of prostate lesions was higher in animals from groups II than in those from groups I, mainly in PCa+Sed II animals when compared with PCa+Sed I (p 0.05). Conclusions: Overall, the animals sacrificed at 61 weeks of age developed more dorsolateral prostate lesions than ani- mals sacrificed at 35 weeks of age, which may be related to a longer testosterone exposure

A Novel Plant-Based Protein Has Similar Effects Compared to Whey Protein on Body Composition, Strength, Power, and Aerobic Performance in Professional and Semi-Professional Futsal Players

IntroductionThe effects of dietary protein on body composition and physical performance seemingly depend on the essential amino acid profile of the given protein source, although controversy exists about whether animal protein sources may possess additional anabolic properties to plant-based protein sources. PurposeTo compare the effects of a novel plant-based protein matrix and whey protein supplementation on body composition, strength, power, and endurance performance of trained futsal players. MethodsFifty male futsal players were followed during 8 weeks of supplementation, with 40 completing the study either with plant-based protein (N = 20) or whey protein (N = 20). The following measures were assessed: bone mineral content, lean body mass, and fat mass; muscle thickness of the rectus femoris; total body water; blood glucose, hematocrit, C-reactive protein, aspartate aminotransferase, alanine aminotransferase, creatine kinase, creatinine, and estimated glomerular filtration rate; salivary cortisol; maximal strength and 1-RM testing of the back squat and bench press exercises; muscle power and countermovement jump; VO2max and maximal aerobic speed. Subjects were asked to maintain regular dietary habits and record dietary intake every 4 weeks through 3-day food records. ResultsNo differences in any variable were observed between groups at baseline or pre- to post-intervention. Moreover, no time*group interaction was observed in any of the studied variables, and a time effect was only observed regarding fat mass reduction. ConclusionsSupplementing with either a novel plant-based protein matrix or whey protein did not affect any of the variables assessed in high-level futsal players over 8 wks. These results suggest that whey protein does not possess any unique anabolic properties over and above those of plant-based proteins when equated to an essential amino acid profile in the population studied. Furthermore, when consuming a daily protein intake >1.6 g/kg BW.day(-1), additional protein supplementation does not affect body composition or performance in trained futsal players, regardless of protein type/source

Rat prostate: practical tips for ultrasonographic monitoring

Background: Prostate is the largest accessory gland of the male reproductive tract. The prostate of men over 40 years- old is frequently affected by several pathologies, like benign prostate hyperplasia and cancer. Rats have been used as model to study prostate cancer. This study intended to address the usefulness of ultrasonography for rat prostate monitoring. Materials and Methods: Eight male Wistar Unilever rats were acquired from Charles River Laboratories and main- tained under controlled conditions of temperature, humidity, air system filtration and light/dark cycle. The prostate was evaluated by ultrasonography in awake animals. The animals were restrained by a researcher and placed in supine position. The skin of the inguinal region was shaved using a machine clipper (AESCULAP® GT420 Isis, USA). A real-time scan- ner (Logic P6®, GE, USA) and a 12 MHz linear transducer were used. Acoustic gel (Parker Laboratories Inc., USA) was applied. A complete transverse scan using B mode was per- formed from the cranial to the caudal region of the prostate, and a sagittal scan was performed moving the probe from the right to the left side. Procedures were approved by the Portuguese Ethics Committee (no.021326). Results: Prostate was easily evaluated by ultrasonography in all animals. In the transverse scan, the urinary bladder presents as a round to oval shape filled with urine (anechoic structure) and the prostate lobes were visible around it. The ventral prostate lobes appear as hypoechoic elongated struc- tures (one right and one left) with a hyperechoic capsule, placed ventrally to the urinary bladder. In this scan, the dorsal prostate was observed close to the urinary bladder neck, as a round hypoechoic structure with a hyperechoic capsule, dor- sally to the urinary bladder. In the sagittal scan, the urinary bladder was observed as an elongated structure filled with urine (anechoic content). The ventral prostate lobes were occasionally observed ventrally to the neck of the urinary bladder, as previously described. The dorsal prostate was ob- served dorsally to the neck of the urinary bladder, presenting as a round to elongated shape, with a hypoechoic appearance and a hyperechoic capsule.Conclusions: The ultrasonography is a non-invasive and ac- cessible tool for prostate monitoring in the rat model. Acknowledgments: This work was supported by European Investment Funds by FEDER/ COMPETE/POCI - Operational Competitiveness and Internationalization Program and National Funds by FCT - Portuguese Foundation for Science and Technology, under the projects Project RUNawayPCa (POCI-01-0145-FEDER-016728 and PTDC/DTP-DES/6077/2014), UIDB/04033/2020, UIDB/ CVT/00772/2020 and UIDB/50006/2020 (LAQV)

Sarcoidose multissistémica: quando somos nós os primeiros a suspeitar!

A Sarcoidose é uma doença inflamatória crónica, granulomatosa, de envolvimento multissistémico e de etiologia desconhecida, que apresenta um espectro alargado de manifestações clínicas. Entre as mais frequentes, destaca-se o envolvimento pulmonar, cutâneo e oftalmológico. Através de um relato de um caso clínico pretende-se discutir as principais manifestações oftalmológicas, o pleomorfismo desta doença e a importância de um diagnóstico precoce. MATERIAL E MÉTODOS: Descrição e discussão de um caso clínico. Doente do sexo feminino, 58 anos, observada no S.U. de Oftalmologia por diminuição de acuidade visual (AV) no olho direito (OD) com cerca de uma semana de evolução. A AV era de 5/10 no OD e de 6/10 no olho esquerdo, apresentando bilateralmente reacção de câmara anterior com precipitados queráticos endoteliais, vitrite e, na fundoscopia, tortuosidade e engurgitamento venosos com periflebite e lesões em “pingos de cera”. Apresentava ainda edema da papila no OD. Concomitantemente referia no último mês aparecimento de nódulos cutâneos nos membros inferiores (MI), queixas álgicas articulares, tosse seca e episódios de febre intermitente acompanhados por suores nocturnos. Encaminhou-se a doente para observação pela Medicina Interna. Foi pedida avaliação laboratorial e imagiológica, bem como biopsia de lesão nodular dos MI. Foram ainda realizados Retinografia, Angiografia Fluouresceínica e Tomografia de Coerência Óptica. RESULTADOS: A avaliação clínica, laboratorial e imagiológica desta doente revelou aspectos sugestivos de Sarcoidose com afecção multissistémica, incluíndo uma panuveíte bilateral com compromisso visual. CONCLUSÃO: A Sarcoidose é uma patologia multiorgânica com frequente envolvimento ocular, sendo que as manifestações oftalmológicas podem constituir a forma de apresentação da doença ou fornecer pistas úteis para o diagnóstico. O diagnóstico precoce é essencial para a correcta orientação do doente e para um tratamento adequado e atempado, minimizando potenciais complicações graves, oculares e sistémicas, que podem advir desta patologia. REFERÊNCIAS BIBLIOGRÁFICAS: Carl P. Herbort et al, “International Criteria for the Diagnosis of Ocular Sarcoidosis: Results of the First International Workshop on Ocular sarcoidosis”, Ocular Immunology and Inflammation, 17, 160-169, 2009 Hilario Nunes et al, Sarcoidosis- Review , Orphanet Journal of Rare diseases 2007, 2:46 Costa F., Arrobas A., Sarcoidose Extratorácica, Artigo de Revisão, Revista Portuguesa de Pneumologia, Vol XIV nº1, Jan/Fev 200

Oclusão da artéria central da retina e oxigenoterapia hiperbárica: a propósito de um caso clínico

Introdução: A Oclusão da Artéria Central da Retina (OACR) constituí uma causa de diminuição súbita, indolor e profunda da acuidade visual. É uma emergência oftalmológica, no entanto, as opções terapêuticas são limitadas. A oxigenoterapia hiperbárica (OH) surge como uma opção terapêutica quando iniciada precocemente. Objectivo: Avaliar o resultado do tratamento de um doente com OACR submetido a oxigenoterapia hiperbárica (OH) instituída nas primeiras 24 horas. Metodos: Descrição de um caso clínico Doente, sexo feminino, 75 anos, com antecedentes pessoais de HTA e hipercolesterolémia, surge no serviço de urgência com quadro de OACR do OE com cerca de 4h de evolução, apresentando à entrada AV OD-0,5 (catarata nuclear) e OE-MM. Foi instituído de imediato tratamento com massagem ocular com lente de 3 espelhos e acetazolamida oral, tendo sido referenciada ao centro de Medicina Hiperbárica do Hospital da Marinha em Lisboa para início de OH. A primeira de 36 sessões teve início às 24h de evolução. Como exames complementares realizou Retinografia, Angiografia Fluoresceínica (AF) e OCT macular (à entrada e após término das 36 sessões), bem como estudo analítico e imagiológico para avaliação de patologia cardiovascular associada. A doente foi referenciada para consulta de Medicina Interna. Resultados: Apresentam-se os resultados funcionais e estruturais oftalmológicos na altura da apresentação, bem como durante e no final do tratamento de OH. Ocorreu uma melhoria da A.V do OE, desde MM na altura da apresentação para 3/10 após 20 sessões, resultado que se manteve no final das 36 sessões. Conclusão: Concluímos que, no caso clínico de OACR apresentado, a implementação precoce de OH contribuíu de forma importante para uma melhoria funcional significativa. A oxigenoterapia a altas pressões parece ser benéfica em situações de doença oclusiva arterial retiniana, possibilitando a oxigenação da retina através do leito coroideu, até que a recanalização espontânea ocorra. Assim, esta opção terapêutica deverá ser considerada, principalmente nos casos de apresentação precoce

Germline MUTYH (MYH) mutations in Portuguese individuals with multiple colorectal adenomas

Germinal mutations in the base excision repair (BER) gene MUTYH (MYH) have recently been described in association with predisposition to multiple colorectal adenomas and cancer. In contrast to the classic dominant condition of familial adenomatous polyposis (FAP) due to germinal mutations in the APC gene, the MYH polyposis is an autosomal recessive disease. The identification of individuals affected by MYH polyposis brings new and important implications for the diagnostic, screening, genetic counseling, follow up and therapeutic options in these patients. In this study, screening for germinal mutations in the MYH gene was performed in 53 Portuguese individuals with multiple colorectal adenomas or classic adenomatous polyposis, in whom no mutation had been identified in the APC gene. The results revealed the presence of biallelic germline MYH mutations in 21 patients. In addition, we here report 3 mutations (c.340T>C [p.Y114H]; c.503G>A [p.R168H]; and c.1186_1187insGG [p.E396fsX437]) which, to our knowledge, have not been previously describe