Transcriptome analyses of the cortex and white matter of focal cortical dysplasia type II: Insights into pathophysiology and tissue characterization.

Introduction: Focal cortical dysplasia (FCD) is a common cause of pharmacoresistant epilepsy. According to the 2022 International League Against Epilepsy classification, FCD type II is characterized by dysmorphic neurons (IIa and IIb) and may be associated with balloon cells (IIb). We present a multicentric study to evaluate the transcriptomes of the gray and white matters of surgical FCD type II specimens. We aimed to contribute to pathophysiology and tissue characterization. Methods: We investigated FCD II (a and b) and control samples by performing RNA-sequencing followed by immunohistochemical validation employing digital analyses. Results: We found 342 and 399 transcripts differentially expressed in the gray matter of IIa and IIb lesions compared to controls, respectively. Cholesterol biosynthesis was among the main enriched cellular pathways in both IIa and IIb gray matter. Particularly, the genes HMGCS1, HMGCR, and SQLE were upregulated in both type II groups. We also found 12 differentially expressed genes when comparing transcriptomes of IIa and IIb lesions. Only 1 transcript (MTRNR2L12) was significantly upregulated in FCD IIa. The white matter in IIa and IIb lesions showed 2 and 24 transcripts differentially expressed, respectively, compared to controls. No enriched cellular pathways were detected. GPNMB, not previously described in FCD samples, was upregulated in IIb compared to IIa and control groups. Upregulations of cholesterol biosynthesis enzymes and GPNMB genes in FCD groups were immunohistochemically validated. Such enzymes were mainly detected in both dysmorphic and normal neurons, whereas GPNMB was observed only in balloon cells. Discussion: Overall, our study contributed to identifying cortical enrichment of cholesterol biosynthesis in FCD type II, which may correspond to a neuroprotective response to seizures. Moreover, specific analyses in either the gray or the white matter revealed upregulations of MTRNR2L12 and GPNMB, which might be potential neuropathological biomarkers of a cortex chronically exposed to seizures and of balloon cells, respectively.

Benchmarking the proteomic profile of animal models of mesial temporal epilepsy.

OBJECTIVES: We compared the proteomic signatures of the hippocampal lesion induced in three different animal models of mesial temporal lobe epilepsy with hippocampal sclerosis (MTLE+HS): the systemic pilocarpine model (PILO), the intracerebroventricular kainic acid model (KA), and the perforant pathway stimulation model (PPS). METHODS: We used shotgun proteomics to analyze the proteomes and find enriched biological pathways of the dorsal and ventral dentate gyrus (DG) isolated from the hippocampi of the three animal models. We also compared the proteomes obtained in the animal models to that from the DG of patients with pharmacoresistant MTLE+HS. RESULTS: We found that each animal model presents specific profiles of proteomic changes. The PILO model showed responses predominantly related to neuronal excitatory imbalance. The KA model revealed alterations mainly in synaptic activity. The PPS model displayed abnormalities in metabolism and oxidative stress. We also identified common biological pathways enriched in all three models, such as inflammation and immune response, which were also observed in tissue from patients. However, none of the models could recapitulate the profile of molecular changes observed in tissue from patients. SIGNIFICANCE: Our results indicate that each model has its own set of biological responses leading to epilepsy. Thus, it seems that only using a combination of the three models may one replicate more closely the mechanisms underlying MTLE+HS as seen in patients.

Avaliação da função sexual e miccional de homens transexuais / Evaluation of sexual and urinary function of transexual men

A transexualidade trata da mudança dos indivíduos de seu sexo masculino ou feminino identificado no nascimento para viver em sociedade sob sua alternativa de identidade de gênero. Visando essa adequação são realizados procedimentos transexualizadores, no entanto, pouco se conhece sobre a função sexual e urinária após esses processos. Neste sentido, este estudo propôs avaliar as funções sexuais e miccionais de homens transexuais submetidos à terapia hormonal. Participaram do estudo 13 homens transexuais e foram utilizados os questionários: Escala de Desconforto Sexual Feminino (FSDS-R), Índice de função sexual feminina (FSFI), Teste de Três Perguntas sobre Incontinência (3IQ), Protection, Amount, Frequency, Adjustment, Bodyimage (PRAFAB). Foi utilizado o software Excel para entrada dos dados, confecção das tabelas e análise estatística descritiva. No FSDS-R, 10 (76,92%) dos homens trans apresentam-se desconfortáveis sexualmente. A pontuação média da FSFI foi de 14,8 pontos, sugerindo disfunção sexual. No teste de 3IQ, 25% relataram perder urinária, destes, 75% apresentam urgência miccional e 25% apresentam Incontinência Urinária (IU) por esforço. No score total do PRAFAB, 50% apresentaram IU leve e 50% obtiveram IU moderada. Portanto, a avaliação foi positiva para tendência a disfunções sexuais nesta população. No que concerne a função miccional, a minoria dos homens trans manifestaram alterações. (AU)

Transsexuality is about changing individuals of their male or female gender identified at birth to live in society under their alternative gender identity. Aiming at this adaptation, transsexualizing procedures are performed, however, little is known about the sexual and urinary function after these processes. In this sense, this study proposed to evaluate the sexual and urinary functions of transsexual men submitted to hormonal therapy. Thirteen transsexual men participated in the study and the questionnaires were used: Female Sexual Discomfort Scale (FSDS-R), Female Sexual Function Index (FSFI), Three Questions Incontinence Test (3IQ), Protection, Amount, Frequency, Adjustment, Body image (PRAFAB). Excel software was used for data entry, preparation of tables and descriptive statistical analysis. In FSDS-R, 10 (76.92%) of trans men are sexually uncomfortable. The average FSFI score was 14.8 points, suggesting sexual dysfunction. In the 3IQ test, 25% reported losing urine, of these, 75% had urinary urgency and 25% had urinary incontinence (UI) on exertion. In the total PRAFAB score, 50% had a mild UI and 50% had a moderate UI. Therefore, the assessment was positive for a tendency towards sexual dysfunction in this population. Regarding the voiding function, the minority of trans men showed changes. (AU)

ITScan: a web-based analysis tool for Internal Transcribed Spacer (ITS) sequences.

BACKGROUND: Studies on fungal diversity and ecology aim to identify fungi and to investigate their interactions with each other and with the environment. DNA sequence-based tools are essential for these studies because they can speed up the identification process and access greater fungal diversity than traditional methods. The nucleotide sequence encoding for the internal transcribed spacer (ITS) of the nuclear ribosomal RNA has recently been proposed as a standard marker for molecular identification of fungi and evaluation of fungal diversity. However, the analysis of large sets of ITS sequences involves many programs and steps, which makes this task intensive and laborious. FINDINGS: We developed the web-based pipeline ITScan, which automates the analysis of fungal ITS sequences generated either by Sanger or Next Generation Sequencing (NGS) platforms. Validation was performed using datasets containing ca. 2,000 to 40,000 sequences each. CONCLUSIONS: ITScan is an online and user-friendly automated pipeline for fungal diversity analysis and identification based on ITS sequences. It speeds up a process which would otherwise be repetitive and time-consuming for users. The ITScan tool and documentation are available at http://evol.rc.unesp.br:8083/itscan.