Breast Cancer Molecular Subtyping in Practice: A Real-World Study of the APIS Breast Cancer Subtyping Assay in a Consecutive Series of Breast Core Biopsies.

The APIS Breast Cancer Subtyping Kit is an mRNA-based assessment of the seven parameters including three biomarkers routinely assessed in all the newly diagnosed breast cancers (BC), oestrogen receptor (ER), progesterone receptor (PR) and HER-2 and an additional four genes that create a novel proliferation signature, MKI67, PCNA, CCNA2 and KIF23. Taken together, the data are used to produce a molecular subtype for every sample. The kit was evaluated against the current standard protocol of immunohistochemistry (IHC) and/or in situ hybridisation (ISH) in breast cancer patients. The data were presented at the weekly breast multidisciplinary team (MDT) meeting. A total of 98 consecutive cases of pre-operative breast cancer core biopsies and two core biopsies of nodal metastases yielding 100 cases were assessed. IHC and APIS results were available for 100 and 99 cases. ER was concordant in 97% cases, PR was concordant in 89% and HER-2 results were concordant with IHC/ISH in 100% of the cases. Ki-67 IHC was discordant in 3% of cases when compared with MK167 alone but discordant in 24% when compared with the four-gene proliferation signature. In conclusion, our study indicates that the APIS Breast Cancer Subtyping Kit is highly concordant when compared to the results produced for ER/PR/HER-2 by IHC and/or ISH. The assay could play a role in the routine assessment of newly diagnosed breast cancer (BC) specimens.

Yellow fever virus infection in human hepatocyte cells triggers an imbalance in redox homeostasis with increased reactive oxygen species production, oxidative stress, and decreased antioxidant enzymes.

Yellow fever (YF) presents a wide spectrum of severity, with clinical manifestations in humans ranging from febrile and self-limited to fatal cases. Although YF is an old disease for which an effective and safe vaccine exists, little is known about the viral- and host-specific mechanisms that contribute to liver pathology. Several studies have demonstrated that oxidative stress triggered by viral infections contributes to pathogenesis. We evaluated whether yellow fever virus (YFV), when infecting human hepatocytes cells, could trigger an imbalance in redox homeostasis, culminating in oxidative stress. YFV infection resulted in a significant increase in reactive oxygen species (ROS) levels from 2 to 4 days post infection (dpi). When measuring oxidative parameters at 4 dpi, YFV infection caused oxidative damage to lipids, proteins, and DNA, evidenced by an increase in lipid peroxidation/8-isoprostane, carbonyl protein, and 8-hydroxy-2'-deoxyguanosine, respectively. Furthermore, there was a significant reduction in the activity of the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx), in addition to a reduction in the ratio of reduced to oxidized glutathione (GSH/GSSG), indicating a pro-oxidant environment. However, no changes were observed in the enzymatic activity of the enzyme catalase (CAT) or in the gene expression of SOD isoforms (1/2/3), CAT, or GPx. Therefore, our results show that YFV infection generates an imbalance in redox homeostasis, with the overproduction of ROS and depletion of antioxidant enzymes, which induces oxidative damage to cellular constituents. Moreover, as it has been demonstrated that oxidative stress is a conspicuous event in YFV infection, therapeutic strategies based on antioxidant biopharmaceuticals may be new targets for the treatment of YF.

Circulating tumor cells as a predictor and prognostic tool for metastatic clear cell renal carcinoma: An immunocytochemistry and genomic analysis.

BACKGROUND: Treatment of metastatic clear cell renal carcinoma (mccRCC) has changed dramatically over the past 20 years, without improvement in the development of biomarkers. Recently, circulating tumor cells (CTCs) have been validated as a prognostic and predictive tool for many solid tumors. OBJECTIVE: We evaluated CTCs in blood samples obtained from patients diagnosed with mccRCC. Comparisons of CTC counts, protein expression profiling, and DNA mutants were made in relation to overall survival and progression-free survival. METHODS: CTCs were isolated from 10 mL blood samples using the ISET® system (Isolation by SizE of Tumor Cells; Rarecells, France) and counted. Protein expression was evaluated in immunocytochemistry assays. DNA mutations were identified with next generation sequencing (NGS). RESULTS: Blood samples (10 mL) were collected from 12 patients with mccRCC before the start of first-line systemic therapy, and again 30 and 60 days after the start of treatment. All 12 patients had CTCs detected at baseline (median, 1.5 CTCs/mL; range: 0.25-7.75). Patients with CTC counts greater than the median had two or more metastatic sites and exhibited worse progression-free survival (19.7 months) compared to those with CTC counts less than the median (31.1 months). Disease progression was observed in 7/12 patients during the study. Five of these patients had baseline CTC counts greater than the median, one had higher CTC levels at the second blood collection, and one patient had CTCs present at 1 CTC/mL which positively stained for PD-L1, N-cadherin, VEGF, and SETD2. CTC DNA from six patients with worse outcomes was subjected to NGS. However, no conclusions could be made due to the low variant allele frequencies. CONCLUSION: Detection of CTCs in patients with mccRCC receiving first-line treatment is a feasible tool with prognostic potential since increased numbers of CTCs were found to be associated with metastasis and disease progression.

Potential of piperine for neuroprotection in sepsis-associated encephalopathy.

AIMS: Sepsis-associated encephalopathy (SAE) is a common complication that increases mortality and leads to long-term cognitive impairment in sepsis survivors. However, no specific or effective therapy has been identified for this complication. Piperine is an alkaloid known for its anti-inflammatory, antioxidant, and neuroprotective properties, which are important characteristics for treatment of SAE. The objective of this study was to evaluate the neuroprotective effect of piperine on SAE in C57BL/6 mice that underwent cecum ligation and perforation surgery (CLP). MAIN METHODS: C57BL/6 male mice were randomly assigned to groups that underwent SHAM surgery or CLP. Mice in the CLP group were treated with piperine at doses of 20 or 40 mg/kg for short- (5 days) or long-term (10 days) periods after CLP. KEY FINDINGS: Our results revealed that untreated septic animals exhibited increased concentrations of IL-6, TNF, VEGF, MMP-9, TBARS, and NLRP3, and decreased levels of BDNF, sulfhydryl groups, and catalase in the short term. Additionally, the levels of carbonylated proteins and degenerated neuronal cells were increased at both time points. Furthermore, short-term and visuospatial memories were impaired. Piperine treatment reduced MMP-9 activity in the short term and decreased the levels of carbonylated proteins and degenerated neuronal cells in the long term. It also lowered IL-6 and TBARS levels at both time points evaluated. Moreover, piperine increased short-term catalase and long-term BDNF factor levels and improved memory at both time points. SIGNIFICANCE: In conclusion, our data demonstrate that piperine exerts a neuroprotective effect on SAE in animals that have undergone CLP.

Cholesterol redistribution triggered by CYP46A1 gene therapy improves major hallmarks of Niemann-Pick type C disease but is not sufficient to halt neurodegeneration.

Cholesterol 24-hydroxylase (CYP46A1) is an exclusively neuronal cytochrome P450 enzyme responsible for converting cholesterol into 24S-hydroxycholesterol, which serves as the primary pathway for eliminating cholesterol in the brain. We and others have shown that increased activity of CYP46A1 leads to reduced levels of cholesterol and has a positive effect on cognition. Therefore, we hypothesized that CYP46A1 could be a potential therapeutic target in Niemann-Pick type C (NPC) disease, a rare and fatal neurodegenerative disorder, characterized by cholesterol accumulation in endolysosomal compartments. Herein, we show that CYP46A1 ectopic expression, in cellular models of NPC and in Npc1tm(I1061T) mice by adeno-associated virus-mediated gene therapy improved NPC disease phenotype. Amelioration in functional, biochemical, molecular and neuropathological hallmarks of NPC disease were characterized. In vivo, CYP46A1 expression partially prevented weight loss and hepatomegaly, corrected the expression levels of genes involved in cholesterol homeostasis, and promoted a redistribution of brain cholesterol accumulated in late endosomes/lysosomes. Moreover, concomitant with the amelioration of cholesterol metabolism dysregulation, CYP46A1 attenuated microgliosis and lysosomal dysfunction in mouse cerebellum, favoring a pro-resolving phenotype. In vivo CYP46A1 ectopic expression improves important features of NPC disease and may represent a valid therapeutic approach to be used concomitantly with other drugs. However, promoting cholesterol redistribution does not appear to be enough to prevent Purkinje neuronal death in the cerebellum. This indicates that cholesterol buildup in neurons might not be the main cause of neurodegeneration in this human lipidosis.

Long-term e-cigarette aerosol exposure causes pulmonary emphysema in adult female and male mice.

This study aimed to evaluate long-term exposure to conventional cigarette smoke (CC) and electronic cigarette (EC) aerosol in adult male and female C57BL/6 mice. Forty-eight C57BL/6 mice were used, male (n = 24) and female (n = 24), both were divided into three groups: control, CC and EC. The CC and EC groups were exposed to cigarette smoke or electronic cigarette aerosol, respectively, 3 times a day for 60 consecutive days. Afterwards, they were maintained for 60 days without exposure to cigarettes or electronic cigarette aerosol. Both cigarettes promoted an influx of inflammatory cells to the lung in males and females. All animals exposed to CC and EC showed an increase in lipid peroxidation and protein oxidation. There was an increase of IL-6 in males and females exposed to EC. The IL-13 levels were higher in the females exposed to EC and CC. Both sexes exposed to EC and CC presented tissue damage characterized by septal destruction and increased alveolar spaces compared to control. Our results demonstrated that exposure to CC and EC induced pulmonary emphysema in both sexes, and females seem to be more susceptible to EC.

High-fat diet increases mortality and intensifies immunometabolic changes in septic mice.

Immunometabolic changes in the liver and white adipose tissue caused by high-fat (HF) diet intake may worse metabolic adaptation and protection against pathogens in sepsis. We investigate the effect of chronic HF diet (15 weeks) on mortality and immunometabolic responses in female mice after sepsis induced by cecum ligation and perforation (CLP). At week 14, animals were divided into four groups: sham C diet, sepsis C diet (C-Sp), sham HF diet (HF-Sh) and sepsis HF diet (HF-Sp). The surviving animals were euthanized on the 7th day. The HF diet decreased survival rate (58.3% vs. 76.2% C-Sp group), increased serum cytokine storm (IL-6 [1.41 ×; vs. HF-Sh], IL-1ß [1.37 ×; vs. C-Sp], TNF [1.34 ×; vs. C-Sp and 1.72 ×; vs. HF-Sh], IL-17 [1.44 ×; vs. HF-Sh], IL-10 [1.55 ×; vs. C-Sp and 1.41 ×; HF-Sh]), white adipose tissue inflammation (IL-6 [8.7 ×; vs. C-Sp and 2.4 ×; vs. HF-Sh], TNF [5 ×; vs. C-Sp and 1.7 ×; vs. HF-Sh], IL-17 [1.7 ×; vs. C-Sp], IL-10 [7.4 ×; vs. C-Sp and 1.3 ×; vs. HF-Sh]), and modulated lipid metabolism in septic mice. In the HF-Sp group liver's, we observed hepatomegaly, hydropic degeneration, necrosis, an increase in oxidative stress (reduction of CAT activity [-81.7%; vs. HF-Sh]; increase MDA levels [82.8%; vs. HF-Sh], and hepatic IL-6 [1.9 ×; vs. HF-Sh], and TNF [1.3 × %; vs. HF-Sh]) production. Furthermore, we found a decrease in the total number of inflammatory, mononuclear cells, and in the regenerative processes, and binucleated hepatocytes in a HF-Sp group livers. Our results suggested that the organism under metabolic stress of a HF diet during sepsis may worsen the inflammatory landscape and hepatocellular injury and may harm the liver regenerative process.

HPV virus and biomarkers of resistance to chemoradiation in circulating tumor cells from patients with squamous cell carcinoma of the anus.

Localized anal cancer is mostly represented by squamous cell carcinoma of the anus (SCCA) and is cured in ≥80 % of cases by chemoradiation (CRT). Development of techniques for detection/evaluating circulating tumor cells (CTCs) for diagnosis/ prognosis/response to therapy can change the manner we treat/follow SCCA patients. OBJECTIVE: to detect CTCs from patients with SCCA and evaluate the presence of HPV virus, p16 expression and markers related to resistance to CRT (RAD23B/ ERCC1/ TYMS) in CTCs at baseline and after CRT. METHODS: CTCs were isolated/quantified by ISET®, protein expressions were analyzed by immunocytochemistry and HPV DNA was detected by chromogenic in situ hybridization. RESULTS: We enrolled 15 patients: median age was 61 (43-73) years, the majority was women (10/15). CTCs were detected in all patients at baseline (median= 0.4 (0.4-3.33) CTCs/mL) and in 8/9 patients, after CRT (median= 2.33 (0-7.0) CTCs/mL). DNA from HPV was found in CTCs in 14/15 patients (93.33 %) at baseline and in 7/9 (77.7 %) after treatment. At a median follow-up of 22.20 (1.45-38.55) months, three patients expressed ERCC1 in CTCs after treatment, with one of them having disease recurrence. CONCLUSION: We showed that detection of HPV in CTCs from patients with non-metastatic SCCA is feasible and appears to be a sensitive diagnostic method. These results may be clinically useful for better monitoring these patients. However, future larger cohorts may demonstrate whether there is any correlation between the presence of HPV and the expression of screening markers for CRT in SCCA.

Prevalence of and factors associated with unmanageable pain levels in people with knee or hip osteoarthritis: a cross-sectional population-based study.

BACKGROUND: Pain due to knee and / or hip osteoarthritis (HKOA) is the most common symptom for seeking healthcare. Pain interferes on daily activities, social and occupational participation in people with HKOA. The goal of this study is to estimate the prevalence of unmanageable pain levels (UPL) among people with HKOA), characterize this population and identify factors associated with UPL, and compare therapeutic strategies used by people with UPL versus manageable pain levels (MPL). METHODS: We analysed data from the EpiReumaPt study (n = 10,661), that included a representative sample of the Portuguese population. Among these, 1081 participants had a validated diagnosis of HKOA by a rheumatologist.. Sociodemographic, lifestyle and health-related data were collected in a structured interview. Pain intensity (NPRS) data were collected in a medical appointment. Painmedication (last month), physiotherapy and surgery were considered as therapies for pain management. UPL was defined as a mean pain intensity in the previous week of ≥5 points on 11-point numeric pain rating scale. The factors associated with UPL were analyzed with logistic regression (p < 0.05, 95%CI). The effect of unmanageable pain levels was assessed by the HOOS/KOOS activities of daily living and quality of life subscales. Symptoms of anxiety and depression were assessed by the Hospital Anxiety and Depression Scale (HADS). Analysis was completed with linear and logistic regression. All analysis were weighted. RESULTS: The estimated prevalence of UPL among people with HKOA was 68.8%. UPL was associated with being female (odds ratio (OR) = 2.36, p < 0.001), being overweight (OR = 1.84, p = 0.035) or obese (OR = 2.26, p = 0.006), and having multimorbidity (OR = 2.08, p = 0.002). People with UPL reported worse performance in activities of daily living and lower quality of life (ß = - 21.28, p < 0.001 and ß = - 21.19, p < 0.001, respectively) than people with MPL. People with UPL consumed more NSAIDs (22.0%, p = 0.003), opioids (4.8%, p = 0.008), paracetamol (2.7%, p = 0.033), and overall analgesics (7.3%, p = 0.013) than people with MPL. A higher proportion of people with UPL underwent physiotherapy (17.5%, p = 0.002) than people with MPL. CONCLUSION: Two-thirds of people with HKOA in Portugal have poor management of their pain levels. Clinical and lifestyle factors, that are highly presented in individuals with HKOA, are associated with unmanageable pain. Our results highlighting the need for further research and implementation of effective interventions to improve pain, function and quality of life in people with HKOA.

Comparação do perfil socioeducacional de crianças com Transtorno do Espectro Autista verbais e não verbais / Comparison between the socio-educational profiles of children with verbal and non-verbal Autism Spectrum Disorder

RESUMO Objetivo Comparar o perfil psicoeducacional de crianças com TEA verbais e não verbais. Método Estudo transversal, com amostra de 30 crianças (15 verbais e 15 não verbais), entre 2 e 9 anos, aproximadamente, com diagnóstico médico de TEA, realizado em equipe. Para a análise do desenvolvimento, aplicou-se a escala de desenvolvimento do Perfil Psicoeducacional Revisado (PEP-R), sendo os dados analisados quantitativamente, de forma comparativa e com uso de testes estatísticos, pelo Modelo ANCOVA para análise de covariância, que avaliou a compatibilidade entre os grupos quanto as pontuações obtidas em cada área tendo como covariável a idade cronológica e o Teste t de Student para Amostras Independentes (nível de significância p ≤ 0,001). Resultados a pontuação nas diferentes áreas do PEP-R foi superior no grupo verbal, havendo relação, na amostra estudada, entre desenvolvimento da linguagem e de habilidades cognitivas e sócio adaptativas. A comparação entre os grupos indica que o perfil do grupo não verbal se encontra rebaixado, com diferenças estatisticamente significantes nas áreas de imitação, percepção, coordenação motora ampla e fina, integração olho mão, desenvolvimento cognitivo e capacidade cognitiva verbal. Conclusão o objetivo de comparar o perfil psicoeducacional de autistas verbais e não verbais foi atingido, apontando diferenças significativas. O perfil dos indivíduos com TEA não verbais analisados na amostra se encontra rebaixado em relação aos verbais. Sugerem-se novos estudos com amostras maiores, faixas etárias delimitadas e com mais testes específicos em cada área do desenvolvimento.

ABSTRACT Purpose Compare the psychoeducational profiles of children with verbal and non-verbal Autism Spectrum Disorder (ASD). Methods Cross-sectional study conducted with a sample of 30 children with a medical diagnosis of ASD (15 verbal and 15 non-verbal) aged 2-9 years. The Psychoeducational Profile-Revised (PEP-R) scale was applied to assess the children's development. The data were analyzed quantitatively and comparatively. Analysis of covariance (ANCOVA) was performed to evaluate the compatibility between the groups regarding the scores obtained in each PEP-R area, with chronological age as the covariate, and Student's t-Test was used for the independent samples (p≤0.001). Results The scores in the different areas of the PEP-R were higher in the verbal group, with associations between language development and cognitive and social adaptive skills in the studied sample. Comparison between the groups showed a lower profile of the non-verbal group, with statistically significant differences in the areas of imitation, perception, gross and fine motor coordination, eye-hand coordination, cognitive performance, and verbal performance. Conclusion The goal of comparing the psychoeducational profiles of verbal and non-verbal ASD children was reached, and statistically significant differences were observed. The children with non-verbal ASD presented a lower psychoeducational profile compared with that of verbal ASD children. Further studies with larger samples, delimited age groups, and more specific tests in each developmental area are suggested.

Enzyme Promiscuity in Serotonin Biosynthesis, From Bacteria to Plants and Humans.

Serotonin is a phylogenetically ancient compound found in animals, plants, and some bacteria. In eukaryotes, serotonin is synthesized from the aromatic amino acid tryptophan via the key enzymes aromatic amino acid hydroxylase (AAAH) and aromatic amino acid decarboxylase (AAAD). Serotonin is also an intermediate in the melatonin biosynthetic pathway and is involved in several vital functions. In humans, serotonin is produced in the gut and in the brain, is critical in the regulation of multiple body functions, and its depletion has been implicated in multiple neurological disorders including depression and Alzheimer's disease, as well as other peripheral conditions namely irritable bowel syndrome and fibromyalgia. The serotonin biosynthetic pathway is well described in eukaryotes, but very little is known about this pathway in bacteria. Evidence points to similar pathways since eukaryote-like AAAH and AAAD (and their genes) have been identified in multiple bacteria, even though serotonin production has not yet been detected in most species. Although data on bacterial tryptophan decarboxylase genes are very limited and no bacterial tryptophan hydroxylase genes have been identified to date, evidence suggests that serotonin production in bacteria might occur through different AAAH and AAAD. Substrate promiscuity in these enzymes has been previously reported and seems to be the key aspect in bacterial serotonin synthesis. Considering the human gut microbiota as a potential source of serotonin, further investigation on its biosynthetic pathways in microbes might lead to important discoveries, which may ultimately foster the development of new therapeutic strategies to treat serotonin depletion-related disorders in humans.

Abdominal wall endometriosis: Report of 83 cases.

OBJECTIVE: To investigate the clinical course and management of abdominal wall endometriosis (AWE). METHODS: A retrospective study was carried out from January 2010 to December 2020, at Vita-Nações Hospitals, Curitiba, Brazil, in order to evaluate data of patients undergoing surgery for the excision of AWE. RESULTS: 83 women with AWE were included in the study. Umbilical scar endometriosis was found in 26 patients (31.3%), being primary in 20 cases (76.9%) and secondary to a laparoscopic procedure in 6 cases (23.1%). 2 patients had secondary implants outside the umbilicus after laparoscopic surgery. Secondary implant after cesarian section in 55 patients (66.3%). Diagnosis was made by ultrasound in 65 patients (78.3%) and by MRI in the remaining 18 (21.7%). Complete excision of the nodule was carried out and no case of recurrence was registered up to now. CONCLUSIONS: Painful abdominal mass presenting in women, especially with a previous history of abdominal and pelvic surgery, should be suspected of AWE. It occurs most often secondary to obstetric or gynecological surgeries and seems to be related to iatrogenic transfer of the endometrial tissue at the level of the surgical scar. Cesarean scar endometriosis is the most common presentation. Surgical excision including the surrounding fibrotic tissue should be performed.

EPA/DHA and linseed oil have different effects on liver and adipose tissue in rats fed with a high-fat diet.

The incidence of cardiovascular diseases and metabolic disorders has increased worldwide. Clinical and experimental research has shown that the consumption of ω-3 FAs can be beneficial to metabolism in several ways, as they can act on metabolic pathways. Our objective was to evaluate the effect of treatment with linseed oil, a vegetable oil rich in alpha-linolenic acid, and EPA and DHA in different proportions (3:1 EPA:DHA, and 1:3 EPA:DHA), on the metabolic disorders induced by a high-fat diet (20 % lipids) in rats for 2 weeks, after 18 weeks of consumption of a high-fat diet. In 18 weeks, the high-fat diet increased blood glucose, systolic blood pressure, triglyceride concentration in the liver and adipose tissue, and impaired insulin sensibility without interfering in the weight of the animals. All treatments were effective in reducing the deposition of hepatic type III collagen, the proportion of ω-6/ω-3 in the liver and WAT (white adipose tissue), the proportion of area/number of adipocytes, and the gene expression of the ACC, FAS, and CPT1 enzymes. In addition, treatment with EPA and DHA reduced blood glucose, serum TNF-α concentration, amount of liver fat, degree of microsteatosis and type I collagen deposition in the liver, deposition of type I and III collagen in TA, gene expression of the transcription factor SREBP-1c, and increased hepatic binucleation. EPA in major proportion was more effective in reducing the area of adipocytes, hepatic triglyceride concentration, PPAR-α expression, and WAT fat weight. DHA in a major proportion reduced the concentration of MCP1 in WAT. LO treatment did not have any isolated effects. We concluded that EPA and DHA were more effective in treating metabolic damage than treatment with LO, leading to a more favorable metabolic profile.

Condições de vida e saúde de professoras da educação básica pública de Minas Gerais provedoras financeiras de suas famílias durante a pandemia de Covid-19 / Living and health conditions of public basic education teachers, economic providers for their families during the Covid-19 pandemic, in Minas Gerais / Condiciones de vida y salud de los docentes de educación básica pública en Minas Gerais, proveedores económicos de sus familias durante la pandemia de COVID-19

Resumo Este artigo busca analisar diferenças nas condições de vida e saúde das professoras principais provedoras do domicílio em comparação às coprovedoras, durante a pandemia de Covid-19. Trata-se de estudo transversal realizado em 2020, por meio de formulário on-line enviado aos professores da rede estadual de Minas Gerais. A variável dependente foi ser ou não a principal provedora da família (principal provedora vs. coprovedora) e as independentes foram agrupadas em sociodemográficas, ocupacionais, situação de saúde e comportamentos. Analisaram-se dados somente das mulheres e estimou-se a regressão logística. Entre as 12.817 professoras participantes, 47,2% declararam-se principais provedoras. Dentre elas, observou-se predomínio de mulheres mais velhas, que viviam sem companheiro(a), com filhos(as) e, no geral, apresentavam características que retrataram pior condição socioeconômica, maior acúmulo de trabalho e comportamentos menos saudáveis. Os resultados permitiram identificar desvantagens nas condições de vida e saúde das professoras principais provedoras financeiras de suas famílias em comparação às coprovedoras.

Abstract The objective was to analyze differences in living and health conditions of teachers who are the main providers for their household compared to co-providers during the COVID-19 pandemic. The cross-sectional study was carried out in 2020, through an online form sent to teachers of public schools in the state of Minas Gerais, Brazil. The dependent variable was whether or not they were their family's main provider (main provider vs. co-provider) and independent variables were grouped into sociodemographic, occupational, health status and behaviors. Only women's data were analyzed and logistic regression was estimated. Among the 12,817 participating female schoolteachers, 47.2% declared to be the main providers. In this subgroup, there was a predominance of older women, who lived without a partner, with children and, in general, these teachers presented worse socioeconomic conditions, greater accumulation of work and less healthy behaviors. The results of the present study allow to identify disadvantages in living and health conditions of female schoolteachers who are the main financial providers of their families compared to co-providers.

Resumen El objetivo fue analizar las diferencias en las condiciones de vida y salud entre las profesoras que son las principales proveedoras del hogar y las coproveedoras, durante la pandemia de COVID-19. Estudio transversal realizado en 2020, a través de un formulario en línea enviado a profesoras de escuelas públicas del estado de Minas Gerais, Brasil. La variable dependiente fue ser o no la proveedora principal de la familia (proveedora principal versus coproveedora) y las variables independientes se agruparon en sociodemográficas, ocupacionales, sanitarias y conductuales. Solo se analizaron los datos de las mujeres y se estimó una regresión logística. Entre las 12.817 maestras participantes, el 47,2 % se declaró proveedora principal. En este subgrupo predominaron las mujeres mayores, que vivían sin pareja, con hijos y, en general, estas profesoras tenían características que retrataban una peor condición socioeconómica, mayor acumulación de trabajo y conductas menos saludables. Los resultados del presente estudio permiten identificar desventajas en las condiciones de vida y salud de las profesoras de escuela que son las principales proveedoras económicas de sus familias en comparación con las coproveedoras.

Developmental Pathways Underlying Lung Development and Congenital Lung Disorders.

Lung organogenesis is a highly coordinated process governed by a network of conserved signaling pathways that ultimately control patterning, growth, and differentiation. This rigorously regulated developmental process culminates with the formation of a fully functional organ. Conversely, failure to correctly regulate this intricate series of events results in severe abnormalities that may compromise postnatal survival or affect/disrupt lung function through early life and adulthood. Conditions like congenital pulmonary airway malformation, bronchopulmonary sequestration, bronchogenic cysts, and congenital diaphragmatic hernia display unique forms of lung abnormalities. The etiology of these disorders is not yet completely understood; however, specific developmental pathways have already been reported as deregulated. In this sense, this review focuses on the molecular mechanisms that contribute to normal/abnormal lung growth and development and their impact on postnatal survival.

Molecular and Dynamic Evaluation of Proteins Related to Resistance to Neoadjuvant Treatment with Chemoradiotherapy in Circulating Tumor Cells of Patients with Locally Advanced Rectal Cancer.

The heterogeneity of response to neoadjuvant chemoradiotherapy (NCRT) is still a challenge in locally advanced rectal cancer (LARC). The evaluation of thymidylate synthase (TYMS) and RAD23 homolog B (RAD23B) expression in circulating tumor cells (CTCs) provides complementary clinical information. CTCs were prospectively evaluated in 166 blood samples (63 patients) with LARC undergoing NCRT. The primary objective was to verify if the absence of RAD23B/TYMS in CTCs would correlate with pathological complete response (pCR). Secondary objectives were to correlate CTC kinetics before (C1)/after NCRT (C2), in addition to the expression of transforming growth factor-ß receptor I (TGF-ßRI) with survival rates. CTCs were isolated by ISET and evaluated by immunocytochemistry (protein expression). At C1, RAD23B was detected in 54.1% of patients with no pCR and its absence in 91.7% of patients with pCR (p = 0.014); TYMS- was observed in 90% of patients with pCR and TYMS+ in 51.7% without pCR (p = 0.057). Patients with CTC2 > CTC1 had worse disease-free survival (DFS) (p = 0.00025) and overall survival (OS) (p = 0.0036) compared with those with CTC2 ≤ CTC1. TGF-ßRI expression in any time correlated with worse DFS (p = 0.059). To conclude, RAD23B/TYMS and CTC kinetics may facilitate the personalized treatment of LARC.

Sesquiterpene lactones from Lychnophora species: Antinociceptive, anti-inflammatory, and antioxidant pathways to treat acute gout.

ETHNOPHARMACOLOGICAL RELEVANCE: Lychnophora trichocarpha and Lychnophora passerina are species used in folk medicine to treat inflammation, pain, and rheumatism. Previous studies have demonstrated the anti-inflammatory effect of ethanol extracts of these species and identified that sesquiterpene lactones contribute to this activity. AIM OF THE STUDY: Gout is an acute inflammatory arthritis caused by the deposition of monosodium urate (MSU) crystals in joints. Inflammation in joints induces oxidative stress in defense cells, releasing pro-inflammatory mediators. This study has three objectives: (1) to demonstrate the effects of sesquiterpene lactones lychnopholide and eremantholide C isolated from L. trichocarpha and goyazensolide isolated from L. passerina on arthritis induced by MSU crystals in C57BL6 mice; (2) to determine whether or not these compounds can inhibit the migration of neutrophils and the release of TNF-α and IL-1ß cytokines in the inflammation region; and (3) to evaluate the effects of sesquiterpene lactones on the activities of the antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) in the cartilage of C57BL/6 mice with gouty arthritis. MATERIALS AND METHODS: The anti-inflammatory, antinociceptive, and antioxidant activities of sesquiterpene lactones in C57BL/6 mice with MSU crystal-induced arthritis were evaluated. In our experimental model, the mice were injected with MSU crystals in the tibiofemoral joint to induce arthritis and then treated with indomethacin, vitamin C, and sesquiterpene lactones. Nociception was evaluated before and after inflammation induction and treatments, neutrophil migration, IL-1ß and TNF-α concentrations, and SOD and CAT activities. RESULTS: Sesquiterpene lactones exerted an anti-inflammatory effect by inhibiting neutrophil migration and TNF-α production. These compounds also demonstrated antinociceptive and antioxidant activities. CONCLUSION: Lychnopholide, eremantholide C, and goyazensolide improved the inflammation induced by MSU crystals by inhibiting the migration of neutrophils to the inflamed area and by blocking the release of the pro-inflammatory cytokine TNF-α. In addition, sesquiterpene lactones reduced oxidative stress by activating SOD and CAT. These results suggest that sesquiterpene lactones have anti-gout activity through the inflammation, pain, and oxidative stress pathways.

Identificação de células tumorais circulantes em pacientes com câncer de canal anal e sua interação com vírus HPV / Identification of circulating tumor cells in patients with anal canal cancer and their interaction with HPV viruses

O câncer tem se tornado um grande problema de saúde pública em países desenvolvidos e em desenvolvimento, sendo responsável por mais de 6 milhões de casos/ óbitos a cada ano, representando cerca de 12% de todas as causas de morte no mundo. O câncer de canal anal corresponde a 4% de todas as neoplasias malignas do trato digestivo baixo e tem como um dos fatores etiológicos, o vírus do papiloma humano (HPV), o que lhe confere forte associação com as doenças sexualmente transmissíveis. A imunossupressão desencadeada pelo HIV também pode acelerar a progressão de lesões precursoras anais associadas ao HPV. Muitos estudos têm demonstrado que as células tumorais circulantes (CTCs) são bons marcadores para avaliação da evolução tumoral. Além disso, essas células são úteis no monitoramento da resposta ao tratamento, contribuindo para avanços na medicina personalizada. O presente estudo teve como objetivos principais: 1) quantificar as CTCs das coletas de primeiro e segundo momento; 2) Correlacionar os achados das análises com evolução clínica dos pacientes. Foram coletadas 24 amostras de sangue total de pacientes com câncer de canal anal localizado e candidatos a tratamento definitivo com quimioradioterapia. Os pacientes tiveram aproximadamente 10 ml de sangue coletados em tubos EDTA em dois momentos distintos: antes do início do tratamento e após seu término. O sangue coletado foi filtrado em uma membrana pelo sistema ISET (Isolation by SizE of Tumor Cells, Rarecells, France). Após a filtração, as células fixadas na membrana de policarbonato foram submetidas à quantificação e à análise por imunocitoquímica (ICC) com os anticorpos p16, CD-45, PD-L1, RAD23B, TYMS e ERCC1 e CISH para detecção da presença do RNAm do vírus HPV. O tempo mediano de seguimento foi de 11 28 meses (1,35- 16,61 meses). Entre as amostras analisadas por ICC, não encontramos nenhuma CTC marcada para p16, houve um paciente cujas CTCs marcaram para PD-L1 na primeira coleta. Dois pacientes tiveram suas CTCs expressando RAD23B na primeira coleta e um na segunda coleta. CTCs expressando TYMS foram encontradas no material de três pacientes na primeira coleta e em um na segunda. ERCC1 estava expresso em CTCs de um paciente na primeira e de três na segunda, onde correlacionou-se com altos níveis de CTCs (p= 0,083) e com ausência de reposta clínica (p= 0,09), porém, sem significância estatística. Nenhuma CTC marcou para CD-45, sendo este anticorpo usado somente para garantir a depleção de leucócitos. Por meio do CISH conseguimos verificar a positividade para HPV nas CTCs em 14 pacientes dos 15 incluídos no estudo na primeira coleta. Os parâmetros obtidos aqui podem ser clinicamente úteis para o melhor monitoramento dos pacientes com câncer de canal anal não metastático, porém, estudos futuros com uma coorte maior precisam ser feitos

Cancer has become a major public health problem in developed and developing countries, accounting for more than 6 million cases of death each year, representing about 12% of all causes of death in the world. Anal canal cancer corresponds to 4% of all malignant neoplasms of the lower digestive tract and has as one of its etiological factors, the human papilloma virus (HPV), which gives it a strong association with sexually transmitted diseases. Immunosuppression triggered by HIV can also accelerate the progression of anal precursor lesions associated with HPV. Many studies have shown that circulating tumor cells (CTCs) are good markers for assessing tumor evolution. In addition, these cells are useful in monitoring the response to treatment, contributing to advances in personalized medicine. The present study had as main objectives: 1) To detect CTCs in patients with cancer of the located anal canal and to identify the presence of the HPV virus in these cells; 2) assess whether there is a relationship between presence of HPV and the number of CTCs, correlating the findings with the clinical evolution of patients through the analysis of medical records. Whole blood samples were collected from patients with cancer of the localized anal canal, candidates for definitive treatment with chemotherapy. The patients had approximately 10 ml of blood collected in EDTA tubes at two different times: before the start of the chemotherapy treatment and after the end of the treatment. The collected blood was filtered on a membrane by the ISET device (Isolation by SizE of Tumor Cells). After filtration, the cells were fixed on the polycarbonate membrane and subjected to quantification and immunocytochemistry (ICC) analysis with antibodies p16, CD-45, PD-L1, RAD23b, TYMS and ERCC1. The median follow-up was 11 28 months (1,35- 16,61 months). There were collected 24 blood samples. Among the samples analyzed by ICC, we did not find any CTC marked for p16. There was one patients whose CTCs stained for PD-L1 in the first collection, two who marked for RAD23B in the first collection and one in the second collection. Three patients had CTCs marked for TYMS in the first collection and one on the second collection. ERCC1 was expressed in CTCs from one patient on the first and in three on the second collection and correlated with higher numbers of CTCs at this moment (p= 0,083) and with absence of clinical response (p= 0,09), although without statistical significance. No CTC stained for CD45, as this antibody was used only to guarantee leukocyte depletion. Through CISH, we verified positivity for HPV in CTCs in 13/15 patients in the first blood collection. The parameters obtained here can be clinically useful for better monitoring of patients with non-metastatic anal canal cancer, however, future studies with a larger cohort need to be done.

Silymarin Attenuates Hepatic and Pancreatic Redox Imbalance Independent of Glycemic Regulation in the Alloxan-induced Diabetic Rat Model.

OBJECTIVE: To evaluate the efficiency of silymarin (SMN) in modulating metabolic parameters and redox status in rats with type 1 diabetes mellitus (T1DM). METHODS: Diabetes was induced by intraperitoneal injection of alloxan. The diabetic rats were administered with SMN at doses of 50 and 100 mg/kg body weight/d for 30 consecutive days. The rats were divided into the following four groups: vehicle control, diabetic (alloxan-treated), DS50 (alloxan + 50 mg/kg body weight/d of SMN), and DS100 (alloxan + 100 mg/kg body weight/d of SMN) groups. The bodyweight and food and water intake were evaluated. After 30 d, the animals were euthanized and the blood was collected for measuring the serum levels of glucose, triacylglycerol (TAG), urea, and creatinine. The liver and pancreas were collected for measuring the activities of superoxide dismutase (SOD) and catalase (CAT), and the levels of carbonylated protein (PC). The pancreas sample was also used for histological analysis. RESULTS: SMN reduced hepatic ( P < 0.001) and pancreatic ( P < 0.001) protein damage and creatinine levels ( P = 0.0141) in addition to decreasing food ( P < 0.001) and water intake ( P < 0.001). However, treatment with SMN did not improve beta-cell function or decrease blood glucose levels in diabetic rats. CONCLUSION: SMN improved polyphagia and polydipsia, renal function, and protected the liver and pancreas against protein damage without affecting hyperglycemia in diabetic animals.

Differences of a Single Injection of Botulinum Toxin A between Infantile and Nonaccommodative Esotropia.

PURPOSES: To compare the results of a single injection of botulinum toxin A (BTA) between children with infantile esotropia (IET) and nonaccommodative esotropia (NAET) during the first 2 years. METHODS: Retrospective study that included 23 children with IET and 25 with NAET. At 6 months, 1 and 2 years after treatment, the deviation and stereoacuity were evaluated. RESULTS: At 6 months and 1 year after treatment there was no difference in ocular alignment between the two groups (success criteria were achieved in 36.8% in IET group and 60.0% in NAET at 6 months p = .129, and 57.9% in IET group and 68.0% in NAET group at 1 year p = .352). Two years after treatment, there were statistical differences between motor alignment (IET group 21,1% and NAET group 60.0%, p = .007) and stereoacuity (IET group 40% and NAET group 90%, p = .004) between the two groups. Although side-effects affected most children during the first week (in the first week, overcorrection was present in 16 (84.2%) children with IET, and in 19 (76.0%) children with NAET; and ptosis affected 15 (78.9%) children with IET and 17 (68.0%) children with NAET), at 6 months all the effects have disappeared on both groups. CONCLUSIONS: We recommend BTA as an alternative, but not as definite treatment in IET if the surgeon/parents are not comfortable with an early strabismus surgery; but retreatment or surgery will have to be considered after 1 year. On the contrary, BTA may be a first-line treatment of NAET because it is an easy, safe and has a long-lasting effect.