Asc-Seurat: analytical single-cell Seurat-based web application.

BACKGROUND: Single-cell RNA sequencing (scRNA-seq) has revolutionized the study of transcriptomes, arising as a powerful tool for discovering and characterizing cell types and their developmental trajectories. However, scRNA-seq analysis is complex, requiring a continuous, iterative process to refine the data and uncover relevant biological information. A diversity of tools has been developed to address the multiple aspects of scRNA-seq data analysis. However, an easy-to-use web application capable of conducting all critical steps of scRNA-seq data analysis is still lacking. We present Asc-Seurat, a feature-rich workbench, providing an user-friendly and easy-to-install web application encapsulating tools for an all-encompassing and fluid scRNA-seq data analysis. Asc-Seurat implements functions from the Seurat package for quality control, clustering, and genes differential expression. In addition, Asc-Seurat provides a pseudotime module containing dozens of models for the trajectory inference and a functional annotation module that allows recovering gene annotation and detecting gene ontology enriched terms. We showcase Asc-Seurat's capabilities by analyzing a peripheral blood mononuclear cell dataset. CONCLUSIONS: Asc-Seurat is a comprehensive workbench providing an accessible graphical interface for scRNA-seq analysis by biologists. Asc-Seurat significantly reduces the time and effort required to analyze and interpret the information in scRNA-seq datasets.

The role of interleukins in vitiligo: a systematic review.

Vitiligo is a disorder of the skin that causes depigmentation and asymptomatic macules whose exact cause is still unclear. Although its aetiology is not fully elucidated, the main theory of its pathomechanism is that it is associated with the autoimmune process. There is few summarized information about the role of inflammatory mediators, as interleukins, in vitiligo, so our aim was to present a systematic review of the role of interleukins in vitiligo, focusing on interleukins. In this review, we included all studies assessing interleukin levels in vitiligo patients conducted up to June 2017. Quality assessment of these studies was performed using the Newcastle-Ottawa Scale (NOS). The interleukins mainly involved were IL-2, IL-4, IL-6, IL-10 and IL-17. The studies highlight the crucial role of IL-17 in the onset and progression of the disease, and its synergistic action with IL-2, IL-6 and IL-33. Dysregulated levels of the interleukins were also correlated with the stage of disease, the affected skin surface area, and indicated as the main factor for lymphocyte infiltration found in depigmented regions. These findings illustrate the growing need for new therapies targeting vitiligo and further research into the role of interleukins as an area of particular interest.

Exercise reverses OVA-induced inhibition of glucocorticoid receptor and increases anti-inflammatory cytokines in asthma.

The purpose of this study was to determine the effect of aerobic exercise training (AT) on the expression of glucocorticoid receptors (GR) and anti-inflammatory cytokines in an asthma model. BALB/c mice were divided into groups control (CT; nonsensitized/nontrained), aerobic training (AT; nonsensitized/trained), ovalbumin (OVA; sensitized/not trained), and OVA+AT (sensitized/trained). OVA groups received OVA by inhalation, and the AT groups completed 1, 3, or 7 days of exercise (60 min/session). Expression of GR, IL-4, IL-5, IL-10, IL-1ra, NF-κB, TGF-ß, VEGF, ICAM-1, VCAM-1; eosinophils counting; and airway remodeling (AR) features [airway smooth muscle (ASM) and epithelial thickness and collagen fiber deposition] were quantified. OVA sensitization induced a decrease in the expression of GR and increases in the eosinophil, IL-4, IL-5, NF-κB, TGF-ß, VEGF, ICAM-1, VCAM-1, and AR features (P < 0.05). After 3 days, AT reversed the OVA-induced reduction in the expression of GR, and subsequently induced increases in the expression of IL-10 and IL-1ra (seventh day). In contrast, the eosinophil migration, the expression of NF-κB, IL-4, IL-5, TGF-ß, RANTES, VEGF, ICAM-1, VCAM-1, and the AR features (P < 0.05) were reduced. AT increases the expression of GR and anti-inflammatory cytokines (IL-10 and IL-1ra) and reduces the expression of inflammatory mediators and airway inflammation in an animal model of asthma.

Airway remodeling is reversed by aerobic training in a murine model of chronic asthma.

The aim of this study was to investigate if the aerobic training (AT) reverses airway remodeling (AR) in an asthma model. BALB/c were divided into four groups: control (unsensitized and untrained); ovalbumin (OVA: sensitized and untrained); AT (unsensitized and trained) and OVA + AT. Allergic inflammation was induced with intraperitoneal and OVA inhalation. AT (low intensity; 5×/week; 60 min/session) was performed at 7, 15, and 30 days. Leukocyte counting in the bronchoalveolar lavage fluid; the expression of IL-5, eotaxin, RANTES, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1); AR features (airway smooth muscle, epithelium thickness, collagen and elastic fibers, mucus production); and AR inducers (transforming growing factor-beta, osteopontin, vascular endothelial growth factor). OVA induced an increase in leukocyte airway migration and increased AR features (P < 0.05). After 7 days, AT reversed the OVA-induced eosinophil and macrophage airway migration, the expression of IL-5, eotaxin, RANTES, ICAM-1, VCAM-1, and all AR inducers. However, total reversion of the AR features and inducers and airway inflammation occurred only after 15 days of AT compared with the OVA groups (P < 0.05) and the effects were maintained until the 30th day. AT reverses AR after 15 days and this effect is preceded by the inhibition of leukocyte migration and occurs simultaneously with the reduction in the expression of inflammatory mediators and AR inducers.

Enhancement of sciatic nerve regeneration after vascular endothelial growth factor (VEGF) gene therapy.

AIMS: Recent studies have emphasized the beneficial effects of the vascular endothelial growth factor (VEGF) on neurone survival and Schwann cell proliferation. VEGF is a potent angiogenic factor, and angiogenesis has long been recognized as an important and necessary step during tissue repair. Here, we investigated the effects of VEGF on sciatic nerve regeneration. METHODS: Using light and electron microscopy, we evaluated sciatic nerve regeneration after transection and VEGF gene therapy. We examined the survival of the neurones in the dorsal root ganglia and in lumbar 4 segment of spinal cord. We also evaluated the functional recovery using the sciatic functional index and gastrocnemius muscle weight. In addition, we evaluated the VEGF expression by immunohistochemistry. RESULTS: Fluorescein isothiocyanate-dextran (FITC-dextran) fluorescence of nerves and muscles revealed intense staining in the VEGF-treated group. Quantitative analysis showed that the numbers of myelinated fibres and blood vessels were significantly higher in VEGF-treated animals. VEGF also increased the survival of neurone cell bodies in dorsal root ganglia and in spinal cord. The sciatic functional index and gastrocnemius muscle weight reached significantly higher values in VEGF-treated animals. CONCLUSION: We demonstrate a positive relationship between increased vascularization and enhanced nerve regeneration, indicating that VEGF administration can support and enhance the growth of regenerating nerve fibres, probably through a combination of angiogenic, neurotrophic and neuroprotective effects.

Primary screening for cervical cancer through self sampling.

OBJECTIVE: To evaluate the use of community health agents (CHAs) to instruct women living in poor rural areas in obtaining self-collected cervical samples and compare the high-risk HPV (hrHPV) hybrid capture (HC) results obtained to those for gynecologist-collected samples. METHODS: After a one-day training, CHAs visited sexually active women, instructing each in the use of collection brush and the Universal Collection Medium tube. One week thereafter, a gynecologist collected cervical samples from, and performed colposcopies on, the same women. A single reference lab performed all HCs. RESULTS: 878 women (Age: 15-69 years) participated. Among self-collected samples, hrHPV prevalence was 33.9% (95% CI: 30.8%-37%), compared with 28.6% (95% CI: 27%-30%) among gynecologist-collected samples. However, 9.3% of the patients were HPV HC II-positive in the self-collected sample and HPV HC II-negative in the gynecologist-collected samples (95% CI: 7.38%-11.22%), whereas 4% tested positive in gynecologist-collected samples and negative in self-collected samples (95% CI: 2.7%-5.3%) (P<0.01; kappa=0.7). Of 9 cases of histologically-confirmed, high-grade squamous intraepithelial lesion, self-collected and provider-collected samples missed one each. CONCLUSION: Self-collected vaginal sampling could be made an additional CHA function under existing program conditions, improving access to cervical cancer screening in poor rural settings.

Function and regulation of the acid and neutral trehalases of Mucor rouxii.

Two different trehalose-hydrolysing activities, known as acid or non-regulatory trehalases, and neutral or regulatory trehalases, have been recognised in a number of fungal species. The true role of these apparently redundant hydrolases remained obscure for many years. However, recent evidence suggests that neutral trehalases would be specialised in the mobilisation of cytosolic trehalose, while acid trehalases would only hydrolyse extracellular trehalose. Results obtained with Mucor rouxii, a Zygomycete initially thought to possess only neutral trehalase activity, reinforced this hypothesis. M. rouxii grows efficiently in trehalose as the sole carbon source. Trehalose-grown or carbon-starved cells exhibit a high trehalase activity of optimum pH 4.5, bound to the external surface of the cell wall, in contrast with the neutral (pH 6.5) trehalase, which occurs in the cytosol. Other differences between the neutral and the acid trehalases are the temperature optimum (35 degrees C and 45 degrees C, respectively) and thermal stability (half-life of 2.5 min and 12 min at 45 degrees C, respectively). The neutral trehalase, but not the acid trehalase, is activated in vitro by cAMP-dependent phosphorylation, stimulated by Ca2+, and inhibited by EDTA. It shows maximal activity at germination and decreases as growth proceeds. In contrast the activity of the acid trehalase is totally repressed in glucose-grown cultures and increases upon exhaustion of the carbon source, and is strongly induced by extracellular trehalose.

Enzymes with gelatinolytic activity can be found in Tityus bahiensis and Tityus serrulatus venoms.

Enzymes with gelatinolytic activity were detected in Tityus bahiensis and Tityus serrulatus venom. Their activity was optimal at pH 8.0 in SDS-PAGE-gelatin. They were inhibited by PMSF but not by iodoacetamide, pepstatin or phenantrolin in the assay conditions used. This suggests that these enzymes are serine proteases. The presence of metal ions did not affect the proteolytic activity of these enzymes. Several possible functions may be envisaged for these enzymes: in tissue permeabilization, pancreatitis and toxin processing.

Elastosis and steroid receptors in primary breast cancer.

The association between the presence of both estrogen (ER) and progesterone (PR) receptors, and the grade of elastosis was evaluated in 100 primary breast carcinomas, 37% of which were from menopausal patients. ER levels were higher in post- than in premenopausal women but only when both ER and PR were present. Progesterone receptor levels were independent of menstrual status but the lowest PR positivity in premenopausal patients was observed during the late luteal phase. The extent of elastosis of the 100 tumors was: Grade 0 (absent), 24%; Grade 1 (small or moderate), 49%, and Grade 2 (marked), 27%. The extent of elastosis was not related to the patient's menstrual status. A significant association was observed between marked elastosis and the presence or absence of both ER and PR. No relationship was observed between the ER + PR and ER-PR+ subclasses and the extent of elastosis. Our data suggest that marked elastosis results from the presence of hormone-dependent cells in human breast carcinomas.

A case of association of duplication of the urogenital and intestinal tracts.

Report of the case of a Caucasian girl, presently 3 years old, who has been followed at the Pediatric Service of the Walter Cantidio Hospital (Federal University of Ceará; Fortaleza, Brazil) since she was 15 days old. At birth she presented a duplication of the bladder, urethra, external genitalia, vagina, colon, and rectum. No cardiac anomalies, bone deformities, or other congenital malformations were detected. When she was 16 months old the child underwent corrective surgery, with rectal vault reconstruction, placing a single and continent anal sphincter in its usual position and resection of the longitudinal vaginal septum, forming a single vaginal cavity. The surgery also resulted in a perineum of normal appearance. Presently, the patient is doing well.

[Hepatitis B antigens systems in schistosomiasis mansoni]. / Sistemas antigênicos da hepatite B na esquistossomose mansôncia.

The authors have studied the antigenic systems HBs and HBe and B virus hepatitis in 155 patients with mansonic schistosomiasis and have found: a) 22,5% HBsAg positivity in patients with the hepatosplenic form of the disease which was associated with histologic liver alterations and 8,7% anti-HBs positivity in patients with the hepatointestinal form of the disease which was associated with a preserved liver histology; in controls they have found 1% positivity for HBsAg. b) HBeAg and anti-HBe were of good prognostic value.

Double gene therapy with granulocyte colony-stimulating factor and vascular endothelial growth factor acts synergistically to improve nerve regeneration and functional outcome after sciatic nerve injury in mice.

Peripheral-nerve injuries are a common clinical problem and often result in long-term functional deficits. Reconstruction of peripheral-nerve defects is currently undertaken with nerve autografts. However, there is a limited availability of nerves that can be sacrificed and the functional recovery is never 100% satisfactory. We have previously shown that gene therapy with vascular endothelial growth factor (VEGF) significantly improved nerve regeneration, neuronal survival, and muscle activity. Our hypothesis is that granulocyte colony-stimulating factor (G-CSF) synergizes with VEGF to improve the functional outcome after sciatic nerve transection. The left sciatic nerves and the adjacent muscle groups of adult mice were exposed, and 50 or 100 µg (in 50 µl PBS) of VEGF and/or G-CSF genes was injected locally, just below the sciatic nerve, and transferred by electroporation. The sciatic nerves were transected and placed in an empty polycaprolactone (PCL) nerve guide, leaving a 3-mm gap to challenge nerve regeneration. After 6 weeks, the mice were perfused and the sciatic nerve, the dorsal root ganglion (DRG), the spinal cord and the gastrocnemius muscle were processed for light and transmission electron microscopy. Treated animals showed significant improvement in functional and histological analyses compared with the control group. However, the best results were obtained with the G-CSF+VEGF-treated animals: quantitative analysis of regenerated nerves showed a significant increase in the number of myelinated fibers and blood vessels, and the number of neurons in the DRG and motoneurons in the spinal cord was significantly higher. Motor function also showed that functional recovery occurred earlier in animals receiving G-CSF+VEGF-treatment. The gastrocnemius muscle showed an increase in weight and in the levels of creatine phosphokinase, suggesting an improvement of reinnervation and muscle activity. These results suggest that these two factors acted synergistically and optimized the nerve repair potential, improving regeneration after a transection lesion.

Mesenchymal stem cells in a polycaprolactone conduit enhance median-nerve regeneration, prevent decrease of creatine phosphokinase levels in muscle, and improve functional recovery in mice.

Although the majority of peripheral-nerve regeneration studies are carried out on the sciatic nerve, lesions of the upper extremities are more common in humans and usually lead to significant physical disabilities. The present study was driven by the hypothesis that a combination of strategies, namely grafts of mesenchymal stem cells (MSC) and resorbable polycaprolactone (PCL) conduits would improve median-nerve regeneration after transection. Mouse median nerves were transected and sutured to PCL tubes that were filled with either green fluorescent protein (GFP(+)) MSC in DMEM or with DMEM alone. During the post-operative period, animals were tested weekly for flexor digitorum muscle function by means of the grasping test. After 8 weeks, the proximal and middle portions of the PCL tube and the regenerating nerves were harvested and processed for light and electron microscopy. The flexor digitorum muscle was weighed and subjected to biochemical analysis for creatine phosphokinase (CK) levels. Scanning electron microscopy of the PCL tube 8 weeks after implantation showed clear signs of wall disintegration. MSC-treated animals showed significantly larger numbers of myelinated and unmyelinated nerve fibers and blood vessels compared with DMEM-treated animals. The flexor digitorum muscle CK levels were significantly higher in the MSC-treated animals, but muscle weight values did not differ between the groups. Compared with the DMEM-treated group, MSC-treated animals showed, by the grasping test, improved functional performance throughout the period analyzed. Immunofluorescence for S-100 and GFP showed, in a few cases, double-labeled cells, suggesting that transplanted cells may occasionally transdifferentiate into Schwann cells. Our data demonstrate that the polycaprolactone conduit filled with MSC is capable of significantly improving the median-nerve regeneration after a traumatic lesion.

Metastase ovariana do carcinoma mamario. Estudo dos ovarios de pacientes ooforectomizadas terapeutica e/ou profilaticamente e necropsiadas. / Ovarian metastasis of mammary carcinoma. Study of patientes submitted to prophylactic and/or therapeutic ooforectomy and with necropsy.

Foram analisados 171 casos de pacientes portadoras de carcinoma mamario submetidos a ooferectomias profilaticas e/ou terapeuticas e a necropsias. Foram 16 (9,4%) os casos com metastases ovarianas. Estes dados foram correlacionados com idade, caracteristicas menstruais, menopausa, bilateralidade, numero de gestacoes, amamentacao e o estadiamento clinico. Os autores chamam a atencao para os beneficios da ooforectomia profilatica no estadio III e que as metastases ovarianas foram mais frequentes nas pacientes no menacme e com ciclos menstruais regulares demonstrando uma maior afinidade das celulas neoplasicas circulantes pelas gonadas funcionantes normais, evidenciando que o fator hormonal se encontra envolvido.}

Estudo mamografico do parenquima mamario em pacientes portadoras do carcinoma de mama. / Mammographic study of the breast parenchymal patients with breast cancer

Foi analisado o comportamento do parenquima mamario atraves da mamografia pelos tipos propostos por Wolfe (N-P1-P2-Dy) em 59 pacientes portadoras de carcinoma mamario.O estudo revelou elevada incidencia para o tipo N, vindo a seguir os tipos P (P1 + P2) e Dy, dados contrarios aos relatos de autores que evidenciaram maior incidencia nos tipos P e Dy, mas concordes com os de outros autores. Os tipos de parenquima foram correlacionados com a idade, paridade, amamentacao, estado menstrual e indice de massa corporal. Os tipos de parenquima P1; P2 e Dy, quando detectados, devem ser considerados como de alto risco, devendo estar sob controle clinico com maior frequencia, apesar de nossos resultados mostrarem o tipo N em 44% dos casos de carcinoma mamario

Determinacao dos receptores estrogenicos (RcE2) no carcinoma mamario. / Determination of the estrogen receptors in breast cancer

Em 27 casos de carcinoma ductal invasor mamario foram determinadas as concentracoes citoplasmicos estrogenicos (RcE2) pela tecnica do carvao revestido com dextrana (CRD) em preparacoes de citosois com positividade de 51,9%. Os casos foram analisados quanto a idade, paridade, amamentacao, estado menstrual, estadiamento clinico e caracteristicas histologicas. Nao houve maior positividade quanto aos parametros referidos, exceto as caracteristicas histologicas, onde a diferenciacao e a celularidade dos tumores foram fatores que demonstraram poder interferir na positividade dos RcE2

Sistemas antigenicos da hepatite B na esquistossomose mansonica. / Hepatitis B antigens systems in schistosomiasis mansoni

Os autores estudaram os sistemas antigenicos HBs e HBe do virus B da hepatite em 155 doentes portadores de esquistossomose mansonica e encontraram: a) 22,5% dos doentes da forma clinica hepato-esplenica foram positivos para o HBsAg mostrando ainda alteracoes histologicas evidentes. 8,7% dos doentes da forma hepato-intestinal mostraram positividade para o anti-HBs e verificou-se preservacao da histologia hepatica. O grupo controle apresentou 1% de positividade para o HBsAg: b) o HBeAg e anti-HBe mostraram ser de valor no prognostico dos casos