Tumor-Infiltrating T Cells in Skin Basal Cell Carcinomas and Squamous Cell Carcinomas: Global Th1 Preponderance with Th17 Enrichment-A Cross-Sectional Study.

Basal cell carcinomas (BCCs) and squamous cell carcinomas (SCCs) are high-incidence, non-melanoma skin cancers (NMSCs). The success of immune-targeted therapies in advanced NMSCs led us to anticipate that NMSCs harbored significant populations of tumor-infiltrating lymphocytes with potential anti-tumor activity. The main aim of this study was to characterize T cells infiltrating NMSCs. Flow cytometry and immunohistochemistry were used to assess, respectively, the proportions and densities of T cell subpopulations in BCCs (n = 118), SCCs (n = 33), and normal skin (NS, n = 30). CD8+ T cells, CD4+ T cell subsets, namely, Th1, Th2, Th17, Th9, and regulatory T cells (Tregs), CD8+ and CD4+ memory T cells, and γδ T cells were compared between NMSCs and NS samples. Remarkably, both BCCs and SCCs featured a significantly higher Th1/Th2 ratio (~four-fold) and an enrichment for Th17 cells. NMSCs also showed a significant enrichment for IFN-γ-producing CD8+T cells, and a depletion of γδ T cells. Using immunohistochemistry, NMSCs featured denser T cell infiltrates (CD4+, CD8+, and Tregs) than NS. Overall, these data favor a Th1-predominant response in BCCs and SCCs, providing support for immune-based treatments in NMSCs. Th17-mediated inflammation may play a role in the progression of NMSCs and thus become a potential therapeutic target in NMSCs.

Corrigendum: Evaluation of PIK3CA mutations in advanced ER+/HER2-breast cancer in Portugal - U-PIK Project.

[This corrects the article DOI: 10.3389/fmolb.2023.1082915.].

Evaluation of PIK3CA mutations in advanced ER+/HER2-breast cancer in Portugal - U-PIK Project.

Background: Around 40% of ER+/HER2-breast carcinomas (BC) present mutations in the PIK3CA gene. Assessment of PIK3CA mutational status is required to identify patients eligible for treatment with PI3Kα inhibitors, with alpelisib currently the only approved tyrosine kinase inhibitor in this setting. U-PIK project aimed to conduct a ring trial to validate and implement the PIK3CA mutation testing in several Portuguese centers, decentralizing it and optimizing its quality at national level. Methods: Eight Tester centers selected two samples of patients with advanced ER+/HER2- BC and generated eight replicates of each (n = 16). PIK3CA mutational status was assessed in two rounds. Six centers used the cobas® PIK3CA mutation test, and two used PCR and Sanger sequencing. In parallel, two reference centers (IPATIMUP and the Portuguese Institute of Oncology [IPO]-Porto) performed PIK3CA mutation testing by NGS in the two rounds. The quality of molecular reports describing the results was also assessed. Testing results and molecular reports were received and analyzed by U-PIK coordinators: IPATIMUP, IPO-Porto, and IPO-Lisboa. Results: Overall, five centers achieved a concordance rate with NGS results (allele frequency [AF] ≥5%) of 100%, one of 94%, one of 93%, and one of 87.5%, considering the overall performance in the two testing rounds. NGS reassessment of discrepancies in the results of the methods used by the Tester centers and the reference centers identified one probable false positive and two mutations with low AF (1-3%, at the analytical sensitivity threshold), interpreted as subclonal variants with heterogeneous representation in the tissue sections processed by the respective centers. The analysis of molecular reports revealed the need to implement the use of appropriate sequence variant nomenclature with the identification of reference sequences (HGVS-nomenclature) and to state the tumor cell content in each sample. Conclusion: The concordance rates between the method used by each tester center and NGS validate the use of the PIK3CA mutational status test performed at these centers in clinical practice in patients with advanced ER+/HER2- BC.

The Aftercare Survey: Assessment and intervention practices after brain tumor surgery in Europe.

Background: People with gliomas need specialized neurosurgical, neuro-oncological, psycho-oncological, and neuropsychological care. The role of language and cognitive recovery and rehabilitation in patients' well-being and resumption of work is crucial, but there are no clear guidelines for the ideal timing and character of assessments and interventions. The goal of the present work was to describe representative (neuro)psychological practices implemented after brain surgery in Europe. Methods: An online survey was addressed to professionals working with individuals after brain surgery. We inquired about the assessments and interventions and the involvement of caregivers. Additionally, we asked about recommendations for an ideal assessment and intervention plan. Results: Thirty-eight European centers completed the survey. Thirty of them offered at least one postsurgical (neuro)psychological assessment, mainly for language and cognition, especially during the early recovery stage and at long term. Twenty-eight of the participating centers offered postsurgical therapies. Patients who stand the highest chances of being included in evaluation and therapy postsurgically are those who underwent awake brain surgery, harbored a low-grade glioma, or showed poor recovery. Nearly half of the respondents offer support programs to caregivers, and all teams recommend them. Treatments differed between those offered to individuals with low-grade glioma vs those with high-grade glioma. The figure of caregiver is not yet fully recognized in the recovery phase. Conclusion: We stress the need for more complete rehabilitation plans, including the emotional and health-related aspects of recovery. In respondents' opinions, assessment and rehabilitation plans should also be individually tailored and goal-directed (eg, professional reinsertion).

Genetic polymorphisms of human platelet antigens in Euro-African and Japanese descendants from Parana, Southern Brazil.

The frequency distributions of HPA-1 to HPA-6 and HPA-15 were evaluated in two Brazilian populations from Parana: a mixed population of predominantly Caucasians and a population of Japanese descendants. Genotyping was performed by PCR-SSP in 364 unrelated individuals. Differences in the distribution of HPA highlight diversity in Brazilian miscegenation and the importance of formation of the HPA panel composed of regional blood donors.

Genetics factors associated with myelodysplastic syndromes.

The myelodysplastic syndromes (MDS) are a clinically and cytogenetically heterogeneous group of clonal diseases. Clonal chromosomal abnormalities are observed in 30-50% of patients with MDS. The deletions are among the most common alterations, and often involve the long arms of chromosomes 5, 7, 8, 13, and 20 and the short arms of chromosomes 12 and 17. The advent of new technologies for the detection of genetic abnormalities led to the description of a new set of recurrent mutations, leading to new insights into the pathophysiology of MDS. The recent recognition that genes involved in the regulation of histone function (EZH2, ASXL1, and UTX) and DNA methylation (DNMT3A, IDH1/IDH2, and TET2) are frequently mutated in MDS, has led to the proposal that there is an important link between genetic and epigenetic alterations in this disease. In fact, regulatory factors have also been considered as miR-143/miR-145, miR-146a, miR-125a and MiR-21. Somatic mutations may influence the clinical phenotype but are not included in current prognostic scoring systems. In recent years research has brought new insights into these diseases, but few of the findings are sufficiently robust to be incorporated into the clinical routine at this time. Thus, the aim of this study was to review the role of genetic factors involved in the diagnosis and development of the different phenotypes of MDS.

Influence of different frequencies of transcutaneous electrical nerve stimulation on the threshold and pain intensity in young subjects.

OBJECTIVE: To investigate the effects of different transcutaneous electrical nerve stimulation frequencies in nociception front of a pressure pain threshold and cold in healthy individuals. METHODS: Twenty healthy subjects were divided into four groups, all of which have gone through all forms of electrical stimulation at different weeks. Assessments were pre and post-therapy, 20 and 60 minutes after stimulation. To evaluate the pressure pain threshold, an algometer was used with one tapered tip, pressing the hypothenar region until voluntary report the word "pain". Cold pain intensity was assessed by immersion in water at 5°C for 30 seconds; at the end, the subject was asked to quantify the pain intensity on a Visual Analog Scale for Pain. For electrical stimulation, two electrodes were used near the elbow, for 20 minutes, with an intensity strong, but not painful. The frequency was in accordance with the group: 0Hz (placebo); 7Hz; 100Hz; and 255Hz. RESULTS: Both for the assessment of pressure pain threshold as the cold pain intensity, there was no significant difference (p>0.05). CONCLUSION: We conclude that the use of transcutaneous electrical nerve stimulation on dermatomes C6 to C8 produced no significant change in pressure pain threshold or cold discomfort.

Influence of different frequencies of transcutaneous electrical nerve stimulation on the threshold and pain intensity in young subjects / As influências de diferentes frequências da estimulação elétrica nervosa transcutânea no limiar e intensidade de dor em indivíduos jovens

Objective To investigate the effects of different transcutaneous electrical nerve stimulation frequencies in nociception front of a pressure pain threshold and cold in healthy individuals. Methods Twenty healthy subjects were divided into four groups, all of which have gone through all forms of electrical stimulation at different weeks. Assessments were pre and post-therapy, 20 and 60 minutes after stimulation. To evaluate the pressure pain threshold, an algometer was used with one tapered tip, pressing the hypothenar region until voluntary report the word “pain”. Cold pain intensity was assessed by immersion in water at 5°C for 30 seconds; at the end, the subject was asked to quantify the pain intensity on a Visual Analog Scale for Pain. For electrical stimulation, two electrodes were used near the elbow, for 20 minutes, with an intensity strong, but not painful. The frequency was in accordance with the group: 0Hz (placebo); 7Hz; 100Hz; and 255Hz. Results Both for the assessment of pressure pain threshold as the cold pain intensity, there was no significant difference (p>0.05). Conclusion We conclude that the use of transcutaneous electrical nerve stimulation on dermatomes C6 to C8 produced no significant change in pressure pain threshold or cold discomfort. .

Objetivo Investigar os efeitos de diferentes frequências da estimulação elétrica nervosa transcutânea na nocicepção, frente a um estímulo doloroso pressórico e ao frio, em indivíduos saudáveis. Métodos Participaram 20 indivíduos saudáveis, divididos em 4 grupos, sendo que todos passaram por todas as formas de eletroestimulação, em semanas diferentes. As avaliações ocorreram nos seguintes períodos: pré-aplicação, pós-aplicação, 20 e 60 minutos após a eletroestimulação. Para avaliar o limiar de dor à pressão, foi utilizado um algômetro com ponta afilada, pressionando na região hipotenar, até o voluntário relatar a palavra “dor”. A intensidade de dor ao frio foi avaliada por meio de imersão em água a 5°C, durante 30 segundos; ao final, pediu-se para que o indivíduo quantificasse a intensidade álgica em uma Escala Visual Analógica de Dor. Para a eletroestimulação, foram utilizados dois eletrodos próximos ao cotovelo, durante 20 minutos, com intensidade referida como forte, porém não dolorosa. A frequência esteve de acordo com o grupo: 0Hz (placebo); 7Hz; 100Hz; e 255Hz. Resultados Tanto para a avaliação do limiar de dor à pressão quanto da intensidade ao frio, não houve diferença significativa (p>0,05). Conclusão O uso da estimulação elétrica nervosa transcutânea, sobre os dermátomos de C6 a C8, não produziu alteração significativa no limiar de dor à pressão e nem no desconforto ao frio. .

Adults with Down syndrome: characterization of a Portuguese sample.

INTRODUCTION: Down syndrome or Trisomy 21 affects physical, behavioral and cognitive development. Ageing people with Down syndrome may suffer from several comorbidities that interfere with their daily life. OBJECTIVE: To analyze in a sample of adults with Down syndrome the presence of common medical diseases, regular use of medication, possible cognitive impairment and oral communication skills. MATERIAL AND METHODS: The socio-demographic and clinical data of 209 Portuguese adults (20-58 years) with Down syndrome were collected. Twenty-six subjects were assessed with the WAIS-III. Data was statistically analyzed--SPSS (v.19). RESULTS: Almost half (47.36%) of the sample suffers from comorbidities, with cardiac, psychiatric, epilepsy and hypothyroidism ranked as the most prevalent diseases. Seventy-two patients (34.45%) take medication that acts on the central nervous system. The WAIS-III results revealed IQ scores between 45-61 (mean = 49.65, SD = 4.93) and 141 individuals (67.5%) who can express themselves through basic oral language. DISCUSSION: In line with other studies, these individuals are vulnerable to various comorbidities. Most have basic oral language, but the IQ values are in the 'extremely low' range, with better performance on verbal tasks. CONCLUSION: The cognitive constraints and communication difficulties in the population with Down syndrome may difficult the early clinical recognition of comorbidities associated with ageing. The identification of new semiology, with caregiver support, is essential to ensure that appropriate care is provided to this population.

Introdução: A síndrome de Down ou Trissomia 21 afeta o desenvolvimento físico, comportamental e cognitivo. No decurso do envelhecimento, a ocorrência de comorbilidades aumenta nesta população, sendo a sua deteção e tratamento precoces determinantes na sua qualidade de vida. Objetivo: Analisar numa amostra de adultos com síndrome de Down a existência de doenças médicas comuns, uso regular de medicação, eventual disfunção cognitiva e capacidades de comunicação oral. Material e Métodos: Foram coletados os dados sociodemográficos e clínicos de 209 adultos portugueses (20 - 58 anos), com síndrome de Down. Vinte e seis indivíduos foram avaliados com a WAIS-III. Os dados foram analisados estatisticamente - SPSS (v.19). Resultados: Quase metade da amostra (47,36%) sofre de comorbilidades, sendo as doenças cardíacas, psiquiátricas, epilepsia e hipotiroidismo as mais prevalentes. Setenta e dois indivíduos (34,45%) tomam medicação com ação no sistema nervoso central. Na WAIS-III, os valores de QI situam-se entre 45-61 (m = 49,65, dp = 4,93) existindo 141 indivíduos (67,5%) capazes de se expressar através de linguagem oral básica. Discussão: Os indivíduos desta amostra são vulneráveis a várias comorbilidades. A maioria tem linguagem oral básica, mas os valores de QI situam-se na faixa 'extremamente baixo', sendo melhor o desempenho em tarefas verbais. Conclusão: As limitações cognitivas e dificuldades comunicacionais na população com síndrome de Down podem condicionar o reconhecimento clínico precoce de comorbilidades que lhe estão associadas no decurso do envelhecimento. A valorização semiológica, com apoio do cuidador, é essencial para garantir que os cuidados de saúde adequados são prestados a esta população.

Atypical phenotype in two patients with LAMA2 mutations.

Congenital muscular dystrophy type 1A is caused by mutations in the LAMA2 gene, which encodes the α2-chain of laminin. We report two patients with partial laminin-α2 deficiency and atypical phenotypes, one with almost exclusive central nervous system involvement (cognitive impairment and refractory epilepsy) and the second with marked cardiac dysfunction, rigid spine syndrome and limb-girdle weakness. Patients underwent clinical, histopathological, imaging and genetic studies. Both cases have two heterozygous LAMA2 variants sharing a potentially pathogenic missense mutation c.2461A>C (p.Thr821Pro) located in exon 18. Brain MRI was instrumental for the diagnosis, since muscular examination and motor achievements were normal in the first patient and there was a severe cardiac involvement in the second. The clinical phenotype of the patients is markedly different which could in part be explained by the different combination of mutations types (two missense versus a missense and a truncating mutation).

Liver and muscle in morbid obesity: the interplay of fatty liver and insulin resistance.

INTRODUCTION: Nonalcoholic fatty liver disease (NAFLD) can be seen as a manifestation of overnutrition. The muscle is a central player in the adaptation to energy overload, and there is an association between fatty-muscle and -liver. We aimed to correlate muscle morphology, mitochondrial function and insulin signaling with NAFLD severity in morbid obese patients. METHODS: Liver and deltoid muscle biopsies were collected during bariatric surgery in NAFLD patients. NAFLD Activity Score and Younossi's classification for nonalcoholic steatohepatitis (NASH) were applied to liver histology. Muscle evaluation included morphology studies, respiratory chain complex I to IV enzyme assays, and analysis of the insulin signaling cascade. A healthy lean control group was included for muscle morphology and mitochondrial function analyses. RESULTS: Fifty one NAFLD patients were included of whom 43% had NASH. Intramyocellular lipids (IMCL) were associated with the presence of NASH (OR 12.5, p<0.001), progressive hepatic inflammation (p = 0.029) and fibrosis severity (p = 0.010). There was a trend to an association between IMCL and decreased Akt phosphorylation (p = 0.059), despite no association with insulin resistance. In turn, hepatic steatosis (p = 0.015) and inflammation (p = 0.013) were associated with decreased Akt phosphoryation. Citrate synthase activity was lower in obese patients (p = 0.047) whereas complex I (p = 0.040) and III (p = 0.036) activities were higher, compared with controls. Finally, in obese patients, complex I activity increased with progressive steatosis (p = 0.049) and with a trend with fibrosis severity (p = 0.056). CONCLUSIONS: In morbid obese patients, presence of IMCL associates with NASH and advanced fibrosis. Muscle mitochondrial dysfunction does not appear to be a major driving force contributing to muscle fat accumulation, insulin resistance or liver disease. Importantly, insulin resistance in muscle might occur at a late point in the insulin signaling cascade and be associated with IMCL and NAFLD severity.

Acquired amyloid neuropathy in a Portuguese patient after domino liver transplantation.

Familial amyloid polyneuropathy (FAP) is a progressive neuropathy with autonomic dysfunction. Domino liver transplantation (DLT), in which the liver of an FAP patient is transplanted into another patient, is routinely applied to compensate for the shortage of available organs. We report a patient who developed a clinical picture of FAP 9 years after a DLT from an FAP donor. Electrophysiological, neuropathological, and autonomic tests were administered. The patient presented with typical clinical features of FAP. Electrophysiological investigation confirmed a moderate sensorimotor axonal and autonomic neuropathy. Sural nerve biopsy confirmed the presence of amyloid deposits in the endoneurium. Skin biopsy at the ankle showed reduced intraepidermal nerve fiber density. Our report shows that FAP can develop in a recipient of an FAP liver. This suggests that careful longitudinal study is required to evaluate the risk of FAP polyneuropathy in patients who undergo domino liver transplantation.

Influence of pupillary diameter, ciliary muscle tone, and ambient light on nerve fiber layer measurements with scanning laser polarimetry.

PURPOSE: To evaluate the influence of the pupillary diameter, ciliary muscle tone, and room light on nerve fiber layer measurements with the scanning laser polarimeter (GDx). METHODS: One randomly selected eye of ten normal volunteers was examined with the GDx in a two-day protocol under eight testing conditions (pilocarpine 1%, phenylephrine 10%, tropicamide 1%, or no drops with room lights on or off). The twelve parameters, obtained by the GDx, were compared under the eight testing conditions, using two way ANOVA for repeated measurements and Tukey HSD post hoc test. RESULTS: Ten of the twelve parameters were statistically significantly different (P < 0.05) when measured under the three medication or no medication conditions, controlling for the ambient light status. There were no significant differences when measured with the light on or off, controlling for use of drops. CONCLUSIONS: Nerve fiber layer measurements with the GDx were influenced by drugs affecting pupillary diameter, but not by the status of room light or ciliary muscle tone.

Enterocolite necrosante: experiência clínico-cirúrgica de 10 anos na Unidade Neonatal da Universidade Federal de Säo Paulo, Escola Paulista de Medicina (Unifesp-EPM) / Necrotizing enterocolitis: clinical and surgical experiency of ten year at Säo Paulo Hospital - Federal University of Säo Paulo

A enterocolite necrosante (ECN) é a afecçäo gastrointestinal adquirida mais frequentemente no período neonatal, acometendo preferencialmente os neonatos prematuros. O objetivo deste estudo reetrospectivo é analisar a evoluçäo clínico-cirúrgica dos recém-nascidos (RNs) que apresentaram ECN na Unidade Neonatal do Hospital Säo Paulo/Escola Paulista de Medicina, entre janeiro de 1986 e dezembro de 1996. Nesse período houve 14.260 nascidos vivos, dos quais 62 RNs tiveram diagnóstico de ECN (0,43 por cento), sendo 52(por cento) (32/62) do sexo feminino e 48 (por cento) (30/62) do masculino. A idade gestacional média foi de 33,4 semanas e a média do peso de nascimento, de 1.622 gramas. Desse total 24 (por cento) dos pacientes (15/62) eram portadores de malformaçöes cardíacas e 27 (por cento) (16/62) apresentaram asfixia perinatal ao nascimento. O tratamento foi exclusivamente clínico em 72,5 (por cento) dos casos (45/62). Baseados nesses dados, pretendemos estabelecer um protocolo prospectivo de intervençäo diagnóstica e terapêutica com o objetivo de obter maior sobrevidaa e menor morbidade dessa afecçäo no período neonatal, além de estimular a aplicaçäo de medidas preventivas, como o uso do leite humano