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Abstract Background Cell therapy has emerged as a revolutionary tool to repair damaged tissues by restoration of an adequate vasculature. Dental Pulp stem cells (DPSC), due to their easy biological access, ex vivo properties, and ability to support angiogenesis have been largely explored in regenerative medicine. Methods Here, we tested the capability of Dental Pulp Stem Cell-Conditioned medium (DPSC-CM), produced in normoxic (DPSC-CM Normox) or hypoxic (DPSC-CM Hypox) conditions, to support angiogenesis via their soluble factors. CMs were characterized by a secretome protein array, then used for in vivo and in vitro experiments. In in vivo experiments, DPSC-CMs were associated to an Ultimatrix sponge and injected in nude mice. After excision, Ultimatrix were assayed by immunohistochemistry, electron microscopy and flow cytometry, to evaluate the presence of endothelial, stromal, and immune cells. For in vitro procedures, DPSC-CMs were used on human umbilical-vein endothelial cells (HUVECs), to test their effects on cell adhesion, migration, tube formation, and on their capability to recruit human CD14+ monocytes. Results We found that DPSC-CM Hypox exert stronger pro-angiogenic activities, compared with DPSC-CM Normox, by increasing the frequency of CD31+ endothelial cells, the number of vessels and hemoglobin content in the Ultimatrix sponges. We observed that Utimatrix sponges associated with DPSC-CM Hypox or DPSC-CM Normox shared similar capability to recruit CD45− stromal cells, CD45+ leukocytes, F4/80+ macrophages, CD80+ M1-macrophages and CD206+ M2-macropages. We also observed that DPSC-CM Hypox and DPSC-CM Normox have similar capabilities to support HUVEC adhesion, migration, induction of a pro-angiogenic gene signature and the generation of capillary-like structures, together with the ability to recruit human CD14+ monocytes. Conclusions Our results provide evidence that DPSCs-CM, produced under hypoxic conditions, can be proposed as a tool able to support angiogenesis via macrophage polarization, suggesting its use to overcome the issues and restrictions associated with the use of staminal cells.

During citizen-science expeditions to the Ulu Temburong National Park, Brunei, several individuals were collected of a semi-slug species of the genus Microparmarion that, based on morphology and in-the-field DNA-barcoding, was found to be an undescribed species.In this paper, we describe Microparmarion sallehi Wu, Ezzwan & Hamdani, n. sp., after field centre supervisor Md Salleh Abdullah Bat. We provide details on the external and internal reproductive morphology, the shell and the ecology of the type locality, as well as a diagnosis comparing it with related species. DNA barcodes were generated for five individuals and used for a phylogenetic reconstruction. Microparmarion sallehi sp. n. and M. exquadratus Schilthuizen et al., 2019 so far are the only Bornean species of the genus that live in lowland forest; other species are found in montane forests.

Myotonic dystrophy type 2 (DM2) is caused by CCTG repeat expansions in the CNBP gene, comprising 75 to >11,000 units and featuring extensive mosaicism, making it challenging to sequence fully expanded alleles. To overcome these limitations, we used PCR-free Cas9-mediated nanopore sequencing to characterize CNBP repeat expansions at the single-nucleotide level in nine DM2 patients. The length of normal and expanded alleles can be assessed precisely using this strategy, agreeing with traditional methods, and revealing the degree of mosaicism. We also sequenced an entire ~50 kbp expansion, which has not been achieved previously for DM2 or any other repeat-expansion disorders. Our approach precisely counted the repeats and identified the repeat pattern for both short interrupted and uninterrupted alleles. Interestingly, in the expanded alleles, only two DM2 samples featured the expected pure CCTG repeat pattern, while the other seven presented also TCTG blocks at the 3′ end, which have not been reported before in DM2 patients, but confirmed hereby with orthogonal methods. The demonstrated approach simultaneously determines repeat length, structure/motif, and the extent of somatic mosaicism, promising to improve the molecular diagnosis of DM2 and achieve more accurate genotype–phenotype correlations for the better stratification of DM2 patients in clinical trials.

Contamination by Aspergillus sp. and the accumulation of its mycotoxins in food and beverages have a high impact on human health and food safety. This investigation inquires the ability of brewer’s yeasts discarded after fermentation (brewing fermentation residue, BFR) to synthesize bioactive compounds and to biocontrol Aspergillus sp. BFRs of Saccharomyces cerevisiae MBELGA62 and Pichia kudriavzevii MBELGA61 proved to have bacteriostatic properties and to be efficient in fungal growth reduction, decreasing the growth rate of Aspergillus flavus and Aspergillus parasiticus up to 37.8% and 42.5%, respectively. Fungal mycelium degradation along with absentia of conidia was detected near the yeast inoculum. Moreover, the yeasts synthesize volatile bioactive compounds that extend Aspergillus sp. lag phase above 100% and decrease fungal growth rates from 20% towards 44%, along with the complete inhibition of conidia synthesis. These results indicate the potential of this residue to be used in biocontrol applications in the food industry.

Abstract Background Alström syndrome is a rare recessively inherited disorder caused by variants in the ALMS1 gene. It is characterized by multiple organ dysfunction, including cone-rod retinal dystrophy, dilated cardiomyopathy, hearing loss, obesity, insulin resistance, hyperinsulinemia, type 2 diabetes mellitus and systemic fibrosis. Heterogeneity and age-dependent development of clinical manifestations make it difficult to obtain a clear diagnosis, especially in pediatric patients. Case presentation Here we report the case of a girl with Alström syndrome. Genetic examination was proposed at age 22 months when suspected macular degeneration was the only major finding. Next generation sequencing of a panel of genes linked to eye-related pathologies revealed two compound heterozygous variants in the ALMS1 gene. Frameshift variants c.1196_1202del, p.(Thr399Lysfs*11), rs761292021 and c.11310_11313del, (p.Glu3771Trpfs*18), rs747272625 were detected in exons 5 and 16, respectively. Both variants cause frameshifts and generation of a premature stop-codon that probably leads to mRNA nonsense-mediated decay. Validation and segregation of ALMS1 variants were confirmed by Sanger sequencing. Conclusions Genetic testing makes it possible, even in childhood, to increase the number of correct diagnoses of patients who have ambiguous phenotypes caused by rare genetic variants. The development of high-throughput sequencing technologies offers an exceptionally valuable screening tool for clear genetic diagnoses and ensures early multidisciplinary management and treatment of the emerging symptoms.

Complete and accurate identification of genetic variants associated with specific phenotypes can be challenging when there is a high level of genomic divergence between individuals in a study and the corresponding reference genome. We have applied the Cas9-mediated enrichment coupled to nanopore sequencing to perform a targeted de novo assembly and accurately reconstruct a genomic region of interest. This approach was used to reconstruct a 250-kbp target region on chromosome 5 of the common bean genome (Phaseolus vulgaris) associated with the shattering phenotype. Comparing a non-shattering cultivar (Midas) with the reference genome revealed many single-nucleotide variants and structural variants in this region. We cut five 50-kbp tiled sub-regions of Midas genomic DNA using Cas9, followed by sequencing on a MinION device and de novo assembly, generating a single contig spanning the whole 250-kbp region. This assembly increased the number of Illumina reads mapping to genes in the region, improving their genotypability for downstream analysis. The Cas9 tiling approach for target enrichment and sequencing is a valuable alternative to whole-genome sequencing for the assembly of ultra-long regions of interest, improving the accuracy of downstream genotype–phenotype association analysis.

Programa de Iniciação à Docência (PID) stimulates the development of new active learning methodologies, integrating both teachers and students. In health literature, researchers have questioned traditional pedagogical models, since them can make the students passive to their own learning process. The purpose of this article is to report the development process of a board game, named ‘Caminhando pelo SUS’, to be used as a learning method. The game contains one board, 55 cards, 4 pins and one manual, and the main purpose is to reach the final house on the board. In order to advance in the game, the player must answer correctly the questions related to its content. The game has been proving to be a helpful active learning methodology, since its dynamics can stimulate the students in a autonomous and proactive way through their learning process.

T1AM, a derivative of thyroid hormones, and its major catabolite, TA1, produce effects on memory acquisition in rodents. In the present study, we compared the effects of exogenous T1AM and TA1 on protein belonging to signal transduction pathways, assuming that TA1 may strengthen T1AM’s effects in brain tissue. A hybrid line of cancer cells of mouse neuroblastoma and rat glioma (NG 108-15), as well as a human glioblastoma cell line (U-87 MG) were used. We first characterized the in vitro model by analyzing gene expression of proteins involved in the glutamatergic cascade and cellular uptake of T1AM and TA1. Then, cell viability, glucose consumption, and protein expression were assessed. Both cell lines expressed receptors implicated in glutamatergic pathway, namely Nmdar1, Glur2, and EphB2, but only U-87 MG cells expressed TAAR1. At pharmacological concentrations, T1AM was taken up and catabolized to TA1 and resulted in more cytotoxicity compared to TA1. The major effect, highlighted in both cell lines, albeit on different proteins involved in the glutamatergic signaling, was an increase in phosphorylation, exerted by T1AM but not reproduced by TA1. These findings indicate that, in our in vitro models, T1AM can affect proteins involved in the glutamatergic and other signaling pathways, but these effects are not strengthened by TA1.

Uma vez revelada a complexidade das experiências biográficas de pessoas em situação de rua, é urgente o aumento do número de estudos que qualifiquem a relação prática entre usuárias/os, políticas públicas e a representação do Estado na execução dos serviços de proteção. Para esta discussão, adicionamos as idiossincrasias sócio-históricas acerca do fenômeno da situação de rua no Brasil e a sua relação com o funcionamento do Sistema Único de Assistência Social (SUAS). No intuito da expansão teórico-prática dessa problemática, este estudo objetivou relatar as experiências de trabalho psicossocial dentro do Estágio Profissional de Psicologia em um serviço de abordagem social no município de Porto Alegre, Brasil. Nossos resultados alocam alguns aspectos estruturais, sócio-históricos e biopolíticos como elementos que interferem na execução ótima dos serviços tipificados pelas políticas públicas, marcando a dissonância entre políticas e suas implementações nos serviços

Resumo O objetivo do presente estudo foi conhecer como se constituíram as vivências na família de origem de homens envolvidos em relações conjugais violentas. Trata-se de pesquisa qualitativa exploratória. Foram entrevistados nove homens autores de violência doméstica, respondendo a processos judiciais com base na Lei º 11.340/2006 (Lei Maria da Penha). Utilizou-se análise temática como método de análise de dados, sendo abordados três temas neste trabalho: negligência afetiva, exposição à violência física na infância e figura masculina de referência. Os resultados foram analisados sob a perspectiva da Teoria Bioecológica do Desenvolvimento Humano. Conclui-se que a construção das masculinidades desses homens sofreu influência de suas vivências anteriores, pautadas por modelos parentais severos, abusivos e negligentes. A figura de referência masculina também é vista como ausente e rígida, exercendo controle sobre o microssistema familiar. Essas experiências podem revelar a construção de um modelo no qual o homem acredita que seu papel também deva ser desempenhado por controle e domínio baseados na violência. Desta forma, isso pode ser considerado um fator de risco para o envolvimento em relações conjugais violentas, já que revela a fragilidade na construção do vínculo de confiança e reverbera nos relacionamentos vividos durante a vida adulta. É importante que esses homens tenham espaços de reflexão sobre suas ações em âmbito conjugal, a fim de ressignificar as experiências de abuso sofridas na infância.

The objective of this study is to evaluate the applicability of the finite element method to analyze pressure distribution in the healthy human foot. Images of a foot were captured using computed tomography and converted into a three-dimensional model, which was adjusted with the aid of CAD software. The model was imported into Abaqus software for finite element analysis, considering the different regions of the foot. Observations of displacement, stresses, and pressure distribution demonstrated a biomechanical behavior of the foot consistent with that reported in the existing literature, regarding the regions of peak plantar pressure. These findings demonstrate the feasibility of evaluating the physical and mechanical behavior of the human foot using the finite element method, and can serve as a reference for the study and manufacture of orthotic appliances, prosthetic devices, and insoles. Level of Evidence V; Prognostic Studies; Expert Opinion.

The incidence of cutaneous malignant melanoma has shown a drastic increase over recent decades, and approximately 70% of newly diagnosed melanoma are tumors with a Breslow thickness less or equal to 1 mm. In the literature, there are well-documented rare cases of late metastasis of thin melanoma, and given their growing incidence, it is expected in the future to see more cases of late recurrence. We present a case report of a metastatic cutaneous melanoma 25 years from diagnosis and a review of the literature. A 61-year-old female patient presented with a newly discovered asymptomatic nodule on her thigh. Her relevant past medical history included a completely excided lesion with Breslow 1.4 mm thickness in 1989 for which she was followed up according to the guidelines and subsequently declared cured after 10 years of surveillance. Fine-needle aspiration and cytological analysis of the lesion proved to be a subcutaneous localization of malignant melanoma. The lesion was completely excised, and the patient has remained disease free since her surgery. The aim of this case report is to emphasize that late metastasis remains uncommon but a definitive cure from melanoma cannot always be considered a disease-free interval of 10 years. Physicians should always be aware of previous melanoma diagnosis with newly discovered suspicious lesions. Better patient education could improve the detection of recurrence and secondary melanomas without any need for more frequent follow-up visits and a prolonged follow-up time.