Diabetes and Stroke: Impact of Novel Therapies for the Treatment of Type 2 Diabetes Mellitus.

Type 2 diabetes mellitus (T2DM) is a significant risk factor for stroke. Nevertheless, the evidence supporting stringent glycemic control to reduce macrovascular complications, particularly stroke, is not as clear as for microvascular complications. Presently, risk reduction strategies are based on controlling multiple risk factors, including hypertension, dyslipidemia, glycemia, smoking, and weight. Since 2008, new pharmacological therapies for treating T2DM have been required to undergo trials to ensure their cardiovascular safety. Remarkably, several novel therapies have exhibited protective effects against the combined endpoint of major cardiovascular events. Evidence from these trials, with stroke as a secondary endpoint, along with real-world data, suggests potential benefits in stroke prevention, particularly with glucagon-like peptide 1 receptor agonists. Conversely, the data on sodium-glucose cotransporter type 2 inhibitors remains more controversial. Dipeptidyl peptidase 4 inhibitors appear neutral in stroke prevention. More recent pharmacological therapies still lack significant data on this particular outcome. This article provides a comprehensive review of the evidence on the most recent T2DM therapies for stroke prevention and their impact on clinical practice.

Improving the Nutritional Status of Socially Vulnerable Children in Manaus, Brazilian Amazon, through a Food Supplementation Programme.

Information on the effects of government nutrition programmes provided to socially vulnerable children to improve their nutritional status is scarce. We analysed the effectiveness of a nutritional programme, including food supplementation with infant formula, on the evolution of the weight and height of socially vulnerable children from Manaus in the Brazilian Amazon. This study included 7752 children aged 12-24 months admitted to the programme between 2017 and 2020. Weight and height measurements at admission and every three months thereafter were extracted from the programme database. Weight-for-age, weight-for-height, body mass index-for-age (BMI/A), and height-for-age z-scores were analysed using a multilevel linear regression model, which showed a statistically significant decrease in nutritional deficits toward nutritional recovery at follow-up. The programme's effectiveness was evaluated in 1617 children using a paired analysis comparing data from between 12 and 15 months of age at admission and follow-up after 6-9 months. Children admitted with wasting presented an increase in the BMI/A z-score, whereas children admitted with a risk of being overweight and obese had a statistically significant decrease in the BMI/A z-score. Children admitted with stunted growth also showed increased height-for-age z-scores. The nutrition programme was effective for children experiencing wasting and reducing excess weight.

Recurrent obstruction of biliary stent-in-stent: a heated solution.

Intraductal radiofrequency ablation (RFA) has been used in the management of malignant biliary obstruction and ampullary neoplasms. Some small studies refer to its role in managing benign biliary strictures with some promising results. The complications are not neglectable, namely cholangitis, pancreatitis, bleeding, and perforation, although most of them can be managed conservatively. There are two catheters available. Only the ERLA (EndoLumunal Radiofrequency Ablation, Taewoong Medical) catheter can control temperature and impedance, allowing it to reduce the risk of complications.

Vitamin D and follicular recruitment in the in vitro fertilization cycle.

OBJECTIVE: Vitamin D (VD) is a fat-soluble steroid hormone, synthesized by the skin, most known for its role in bone mineral balance. Vitamin D receptors (VDR) are also found in the female reproductive system, but their role remains unclear. The objective of this study was to analyze the relationship between serum vitamin D levels and the number of oocytes retrieved after ovarian stimulation. METHODS: This is a retrospective study involving 267 patients undergoing in vitro fertilization (IVF) carried out in the Fertipraxis clinic, a private practice facility. The patients were initially divided into two groups according to their VD levels. Group 1 included 152 patients with VD levels < 30 ng/mL and group 2 had 115 patients with VD levels > 30 ng/mL. They were further analyzed and separated considering their age, anthropometric data, ovarian reserve, amount of gonadotropin used, and follicles obtained until trigger day. RESULTS: In our analysis, there were no difference in the number of follicles and oocytes retrieved, nor in the number of mature oocytes obtained from patients with both vitamin D deficiency and sufficiency. CONCLUSIONS: The results of our study show no difference among number of follicles, oocytes retrieved and mature oocytes obtained after ovarian stimulation according to their vitamin D serum levels. Further higher-quality studies are needed to evaluate the possible roles of serum vitamin D levels in other stages of human fertilization process.

Challenges in Extubation: A Case of a Non-removable Tracheal Tube Following Surgery.

This report details a challenging case of difficult extubation due to a lodged tracheal tube following surgery, presenting an unexpected and complex clinical situation. An inspection of the airway using videolaryngoscopy revealed an over-inflated cuff beneath the vocal cords. Initial efforts to deflate the cuff with various methods were unsuccessful. The situation was ultimately resolved through the intervention of an otolaryngology surgeon. This case not only reviews various mechanisms of difficult endotracheal tube removal reported in the literature, but also underscores the potential for serious complications and highlights the critical role of multidisciplinary collaboration in managing extubation challenges. Additionally, our manuscript discusses alternative strategies that can be employed in scenarios where an otolaryngology surgeon is not available, offering practical guidance for anesthesiologists confronted with similar situations.

Effective Postoperative Pain Management in Thoracic Outlet Syndrome Surgery: The Role of the Erector Spinae Plane Block.

Thoracic outlet syndrome (TOS) often necessitates surgical intervention to alleviate neurovascular bundle compression, which can result in severe postoperative pain. The myriad of surgical techniques available for TOS treatment, the intricate involvement of diverse sensory pathways, and the limited literature on effective analgesic methods for these specific cases underscore the need for successful approaches. This report introduces an efficacious multimodal analgesic strategy that incorporates the erector spinae plane (ESP) block to enhance postoperative pain management after a supraclavicular surgical approach. By combining this fascial block with a comprehensive rationale for its implementation, this case offers valuable insights into improving the postoperative care of TOS patients, ultimately aiming to enhance their comfort and recovery.

Safety of Endoscopy Units during the COVID-19 Pandemic.

Introduction: The COVID-19 pandemic drastically changed the daily routine of all healthcare systems worldwide, and endoscopy units were no exception. Endoscopic exams were considered to have a high risk of transmission, and therefore, the safety of endoscopy units and the consequent need for pre-endoscopy SARS-CoV-2 screening were questioned early on. The aim of our study was to assess the safety of endoscopy units during the COVID-19 pandemic, as well as the effectiveness/necessity for SARS-CoV-2 screening prior to endoscopies. Material and Methods: This is a retrospective and single-center study carried out in a Portuguese tertiary hospital. All patients who underwent endoscopic procedures between September 1, 2020 and February 28, 2021 were included. The pre-endoscopy screening consisted of a specific questionnaire or a RT-PCR test for SARS-CoV-2 (nasal and oropharyngeal swab). Data were obtained through patient's clinical records and the Trace COVID platform. Results: A total of 2,166 patients were included. Patients had a mean age of 61.8 years and were predominantly male (56.2%, n = 1,218). Eighty-one (3.7%) patients had previous SARS-CoV-2 infection, with a median difference of 74 days (IQ 40.5:160.5) between infection and endoscopy. Most patients (70.2%, n = 1,521) underwent PCR screening for SARS-CoV-2 up to 72 h before the procedure, with the remaining patients (29.8%, n = 645) answering a questionnaire of symptoms and risk contacts up to 3 days before endoscopy. Of the patients who underwent RT-PCR screening for SARS-CoV-2, 21 (1.4%) tested positive, and all were asymptomatic at the time of the screening. The evaluation for SARS-CoV-2 infection up to 14 days after the endoscopic exams identified 9 positive patients (0.42%) for SARS-CoV-2. The median difference in days between endoscopy and the diagnosis of infection was 10 days. Discussion/Conclusion: Pre-endoscopy screening with RT-PCR test for SARS-CoV-2 identified a very small number of patients with COVID-19 infection as well as patients with COVID-19 infection in the following 14 days. Therefore, the risk of infection in endoscopy units is negligible if screening of symptoms and risk contacts is applied and individual protective equipment is used.

Introdução: A pandemia COVID-19 mudou drasticamente o dia-a-dia de todos os sistemas de saúde a nível mundial e as unidades de endoscopia não foram exceção. Os exames endoscópicos foram considerados exames com alto risco de transmissão pelo que desde cedo se questionou a segurança das unidades de endoscopia e a consequente necessidade de rastreio SARS-CoV-2 pré-endoscopia. O objetivo do estudo foi avaliar a segurança das unidades de endoscopia durante a pandemia por COVID-19 bem como a eficácia/necessidade de rastreio SARS-CoV-2 prévio aos exames endoscópicos. Material e métodos: Foi desenvolvido um estudo retrospetivo e unicêntrico, no qual todos os doentes submetidos a exames endoscópicos entre 1 de setembro de 2020 e 28 de fevereiro de 2021 foram incluídos. Como estratégia de rastreio pré endoscopia foram aplicados questionários específicos de sintomas e contactos de risco, ou teste PCR de SARS-CoV-2 (zaragatoa nasal e orofaríngea). Os dados clínicos foram obtidos através do processo clínico do doente e da plataforma Trace COVID-19. Resultados: Foram incluídos um total de 2,166 doentes submetidos a exames endoscópicos durante o período de estudo. Os doentes incluídos apresentaram uma média de idades de 61.8 anos e eram maioritariamente do sexo masculino (56.2%, n = 1,218). 3.7% (n = 81) dos doentes já tinha tido infeção por COVID-19 no passado, sendo a mediana da diferença de dias entre a infeção e a data do exame de 74 dias. A maioria dos doentes (70.2%, n = 1,521) foi submetido a rastreio por PCR de SARS-CoV-2 até 72 horas antes do procedimento, sendo os restantes doentes (29.8%, n = 645) submetidos a um questionário de sintomas e contactos de risco realizado até 3 dias antes do procedimento. Dos doentes que realizaram rastreio por PCR de SARS-CoV-2, 21 (1.4%) apresentaram teste positivo, estando todos assintomáticos à data do teste. Aquando da verificação de infeção por SARS-CoV-2 até 14 dias após a realização dos exames endoscópicos apurou-se que apenas 9 doentes (0.42%) testaram positivo para SARS-CoV-2, sendo a mediana da diferença de dias entre a data do exame e o diagnóstico de infeção de 10 dias. Discussão/Conclusão: O rastreio pré-endoscopia com teste PCR de SARS-CoV-2 identificou um número reduzido de doentes infetados com COVID-19 e o número de doentes com infeção por COVID-19, nos 14 dias seguintes aos exames endoscópicos, foi muito baixo. Assim, se aplicado o rastreio de sintomas e contactos de risco, usados os equipamentos de proteção individual adequados, o risco de infeção nas unidades de endoscopia torna-se negligenciável.

Postcolonoscopy Colorectal Cancer in a Referral Center for Colorectal Cancer: Prevalence and Risk Factors.

Background and Aims: Colonoscopy is effective to detect and remove colorectal lesions. However, after a negative colonoscopy, cancers could be detected during the interval follow-up. This study was designed to identify characteristics and risk factors for postcolonoscopy colorectal cancer - interval type. Methods: Medical records of individuals who were newly diagnosed with colorectal cancer between January 2018 and December 2019 were reviewed. Clinical, demographic, and endoscopic variables were analyzed. Those with the diagnosis of colorectal cancer between two consecutive colonoscopies performed within the appropriated surveillance range were considered to have postcolonoscopy colorectal cancer - interval type. A comparison between the group of patients with non-postcolonoscopy colorectal cancer - interval type and the group of patients with postcolonoscopy colorectal cancer - interval type was then performed. Results: During the study period, 491 patients were newly diagnosed with colorectal cancer. Among them, 61 (12.4%) had postcolonoscopy colorectal cancer - interval subtype. Postcolonoscopy colorectal cancer - interval type was three times more prevalent on the proximal colon (p = 0.014) and was associated with the presence of two or more cardiovascular risk factors (aOR = 4.25; p = 0.016), cholecystectomy in the past (aOR = 10.09; p = 0.019), and family history of colorectal cancer on a first-degree relative (aOR = 4.25; p = 0.006). Moreover, isolated cardiovascular risk factors revealed a protective effect for the absence of all cardiovascular risk factors (aOR = 20; p = 0.034). The ROC curve associated with the multivariate model revealed a predictive power of 77.8% (p < 0.001). Conclusions: Postcolonoscopy colorectal cancer - interval type is more common in the proximal colon and in patients with a family history (first-degree relative) of colorectal cancer, two or more cardiovascular risk factors, and a history of cholecystectomy. All of these are easily detectable in clinical practice and may be of extreme importance in the control of postcolonoscopy colorectal cancer in the near future.

Introdução: A colonoscopia é eficaz a detetar e remover lesões do colon e reto. Contudo, após uma colonoscopia normal, podem ser detetadas neoplasias durante o intervalo de vigilância recomendado entre colonoscopias. O objetivo do estudo foi identificar características e fatores de risco para o desenvolvimento de cancro colorretal póscolonoscopia ­ subtipo de intervalo. Material e Métodos: Estudo retrospetivo e unicêntrico realizado entre janeiro de 2018 e dezembro de 2019 que incluiu todos os doentes diagnosticados de novo com cancro colorretal. Variáveis clínicas, demográficas e endoscópicas foram obtidas após consulta do processo clínico. Doentes com diagnóstico de cancro colorretal entre duas colonoscopias consecutivas, realizadas no intervalo de vigilância recomendado, foram considerados como tendo cancro colorretal pós-colonoscopia ­ subtipo de intervalo. Foi, então, realizada a comparação entre o grupo de doentes com cancro colorretal não pós colonoscopia ­ subtipo de intervalo e o grupo de doentes com cancro colorretal pós colonoscopia ­ subtipo de intervalo. Resultados: Durante o período de estudo, 491 doentes foram diagnosticados de novo com cancro colorretal. Destes, 61 (12.4%) foram considerados como tendo cancro colorretal pós-colonoscopia ­ subtipo de intervalo. O cancro colorretal pós-colonoscopia ­ subtipo de intervalo foi três vezes mais prevalente no colon proximal (p = 0.014) e associou-se a presença de dois ou mais fatores de risco cardiovasculares (aOR = 0.45; p = 0.016), colecistectomia no passado (aOR = 10.09; p = 0.0.19) e história familiar de cancro colorretal num familiar de primeiro grau (aOR = 4.25; p = 0.006). Aquando da análise dos fatores de risco cardiovasculares isolados, observou- se um fator protetor aquando da ausência de todos os fatores de risco cardiovasculares (aOR = 20; p = 0.034). A curva ROC associada ao modelo multivariado revelou um poder preditivo de 77.8% (p < 0.001). Conclusão: O cancro colorretal pós-colonoscopia ­ subtipo de intervalo é mais comum no colon proximal e em doentes com história familiar (em familiares de primeiro grau) de cancro colorretal, dois ou mais fatores de risco cardio-vasculares e história de colecistectomia. Todos estes fatores de risco são facilmente detetáveis na prática clínica e podem ser de extrema importância no controlo, a curto e longo prazo, do cancro colorretal pós-colonoscopia.

Cross-species oncogenomics offers insight into human muscle-invasive bladder cancer.

BACKGROUND: In humans, muscle-invasive bladder cancer (MIBC) is highly aggressive and associated with a poor prognosis. With a high mutation load and large number of altered genes, strategies to delineate key driver events are necessary. Dogs and cats develop urothelial carcinoma (UC) with histological and clinical similarities to human MIBC. Cattle that graze on bracken fern also develop UC, associated with exposure to the carcinogen ptaquiloside. These species may represent relevant animal models of spontaneous and carcinogen-induced UC that can provide insight into human MIBC. RESULTS: Whole-exome sequencing of domestic canine (n = 87) and feline (n = 23) UC, and comparative analysis with human MIBC reveals a lower mutation rate in animal cases and the absence of APOBEC mutational signatures. A convergence of driver genes (ARID1A, KDM6A, TP53, FAT1, and NRAS) is discovered, along with common focally amplified and deleted genes involved in regulation of the cell cycle and chromatin remodelling. We identify mismatch repair deficiency in a subset of canine and feline UCs with biallelic inactivation of MSH2. Bovine UC (n = 8) is distinctly different; we identify novel mutational signatures which are recapitulated in vitro in human urinary bladder UC cells treated with bracken fern extracts or purified ptaquiloside. CONCLUSION: Canine and feline urinary bladder UC represent relevant models of MIBC in humans, and cross-species analysis can identify evolutionarily conserved driver genes. We characterize mutational signatures in bovine UC associated with bracken fern and ptaquiloside exposure, a human-linked cancer exposure. Our work demonstrates the relevance of cross-species comparative analysis in understanding both human and animal UC.

Metabolomic analysis of follicular fluid from women with Hashimoto thyroiditis.

Hashimoto thyroiditis is an autoimmune disease characterized by hypothyroidism and a high level of anti-thyroid autoantibodies. It has shown to negatively impact female fertility; however, the mechanisms are unclear. Ovarian follicular fluid appears to be the key to understanding how Hashimoto thyroiditis affecst fertility. Thus, we aimed to evaluated the metabolic profile of follicular fluid and antithyroid autoantibody levels in the context of Hashimoto thyroiditis. We collected follicular fluid from 61 patients, namely 38 women with thyroid autoantibody positivity and 23 women as negative controls, undergoing in vitro fertilization treatment. Follicular fluid samples were analyzed using metabolomics, and thyroid autoantibodies were measured. Fifteen metabolites with higher concentrations in the follicular fluid samples from Hashimoto thyroiditis were identified, comprising five possible affected pathways: the glycerophospholipid, arachidonic acid, linoleic acid, alpha-linolenic acid, and sphingolipid metabolism pathways. These pathways are known to regulate ovarian functions. In addition, antithyroglobulin antibody concentrations in both serum and follicular fluid were more than tenfold higher in women with Hashimoto thyroiditis than in controls. Our data showed that the metabolic profile of follicular fluid is altered in women with Hashimoto thyroiditis, suggesting a potential mechanistic explanation for the association of this disease with female infertility.

De Novo Human Angiotensin-Converting Enzyme 2 Decoy NL-CVX1 Protects Mice From Severe Disease After Severe Acute Respiratory Syndrome Coronavirus 2 Infection.

The emergence of novel variants of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) underscores the need to investigate alternative approaches to prevent infection and treat patients with coronavirus disease 2019. Here, we report the preclinical efficacy of NL-CVX1, a de novo decoy that blocks virus entry into cells by binding with nanomolar affinity and high specificity to the receptor-binding domain of the SARS-CoV-2 spike protein. Using a transgenic mouse model of SARS-CoV-2 infection, we showed that a single prophylactic intranasal dose of NL-CVX1 conferred complete protection from severe disease following SARS-CoV-2 infection. Multiple therapeutic administrations of NL-CVX1 also protected mice from succumbing to infection. Finally, we showed that infected mice treated with NL-CVX1 developed both anti-SARS-CoV-2 antibodies and memory T cells and were protected against reinfection a month after treatment. Overall, these observations suggest NL-CVX1 is a promising therapeutic candidate for preventing and treating severe SARS-CoV-2 infections.

Immunostaining of ßA-Activin and Follistatin Is Decreased in HPV(+) Cervical Pre-Neoplastic and Neoplastic Lesions.

The activin-follistatin system regulates several cellular processes, including differentiation and tumorigenesis. We hypothesized that the immunostaining of ßA-activin and follistatin varies in neoplastic cervical lesions. Cervical paraffin-embedded tissues from 162 patients sorted in control (n = 15), cervical intraepithelial neoplasia (CIN) grade 1 (n = 38), CIN2 (n = 37), CIN3 (n = 39), and squamous cell carcinoma (SCC; n = 33) groups were examined for ßA-activin and follistatin immunostaining. Human papillomavirus (HPV) detection and genotyping were performed by PCR and immunohistochemistry. Sixteen samples were inconclusive for HPV detection. In total, 93% of the specimens exhibited HPV positivity, which increased with patient age. The most detected high-risk (HR)-HPV type was HPV16 (41.2%) followed by HPV18 (16%). The immunostaining of cytoplasmatic ßA-activin and follistatin was higher than nuclear immunostaining in all cervical epithelium layers of the CIN1, CIN2, CIN3, and SCC groups. A significant decrease (p < 0.05) in the cytoplasmic and nuclear immunostaining of ßA-activin was detected in all cervical epithelial layers from the control to the CIN1, CIN2, CIN3, and SCC groups. Only nuclear follistatin immunostaining exhibited a significant reduction (p < 0.05) in specific epithelial layers of cervical tissues from CIN1, CIN2, CIN3, and SCC compared to the control. Decreased immunostaining of cervical ßA-activin and follistatin at specific stages of CIN progression suggests that the activin-follistatin system participates in the loss of the differentiation control of pre-neoplastic and neoplastic cervical specimens predominantly positive for HPV.

Combined effects of intermittent fasting with swimming-based high intensity intermittent exercise training in Wistar rats.

High caloric intake and physical inactivity are known precursors to the development of several chronic metabolic diseases. For obesity and sedentarism, High Intensity Intermittent Exercise (HIIE) and Intermittent Fasting (IF) have emerged as individual strategies to attenuate their negative effects by improving metabolism. To study their combined effects, Wistar male rats (n = 74, 60 days old) were divided into four groups: Sedentary Control (C), swimming-based HIIE only (HIIE), Intermittent Fasting only (IF), and swimming-based HIIE associated with Intermittent Fasting (HIIE/IF). Over an eight-week period swimming performance, body composition, weight and feeding behavior were analyzed. The final morphology of white adipose tissue showed a significant reduction in adipocyte size consistent with a higher number of cells per area in exercised animals (vs C and IF, p < 0.05), which also displayed characteristics of browning through UCP-1 levels and CD31 staining. These results suggest that the increased performance in the HIIE/IF group is, in part, by modifications of WAT metabolism through the browning process.

Ultrasound-Guided Electroacupuncture for Thoracic Myofascial Pain Syndrome: A Case Report.

Myofascial pain syndrome is a painful condition characterized by trigger points in muscles that can be treated effectively with acupuncture. While cross-fiber palpation can help localize trigger points, needle accuracy may be limited and accidental puncture of delicate structures, such as the lung, is a risk, as evidenced by reports of pneumothorax after acupuncture. Ultrasound imaging can help in reducing the risk of iatrogenic pneumothorax from needling, but there is a paucity of papers describing the use of ultrasound imaging during acupuncture. We present a report on electroacupuncture treatment for myofascial pain syndrome using real-time ultrasound guidance, aimed at avoiding accidental puncture of the pleura when targeting deep muscle layers in the thoracic region.

UBE2C promotes leptomeningeal dissemination and is a therapeutic target in brain metastatic disease.

Background: Despite current improvements in systemic cancer treatment, brain metastases (BM) remain incurable, and there is an unmet clinical need for effective targeted therapies. Methods: Here, we sought common molecular events in brain metastatic disease. RNA sequencing of thirty human BM identified the upregulation of UBE2C, a gene that ensures the correct transition from metaphase to anaphase, across different primary tumor origins. Results: Tissue microarray analysis of an independent BM patient cohort revealed that high expression of UBE2C was associated with decreased survival. UBE2C-driven orthotopic mouse models developed extensive leptomeningeal dissemination, likely due to increased migration and invasion. Early cancer treatment with dactolisib (dual PI3K/mTOR inhibitor) prevented the development of UBE2C-induced leptomeningeal metastases. Conclusions: Our findings reveal UBE2C as a key player in the development of metastatic brain disease and highlight PI3K/mTOR inhibition as a promising anticancer therapy to prevent late-stage metastatic brain cancer.

Randomized prospective study comparing conventional In Vitro Fertilization technique to Intravaginal Culture with the INVOCELL™ device for 3 and 5 days.

OBJECTIVE: The objective was to analyse and compare the formation and quality of the embryos developed using conventional in vitro fertilization (IVF) and IVC techniques with an INVOCell™ device. METHODS: Two groups were formed, with eight couples in each, one in culture for three days (D3) and another in culture for five days (D5), using intravaginal culture technique with an INVOcell device and a conventional in vitro fertilization technique. RESULTS: Embryo formation in Group D5 showed 46.7% (IVC) and 40% (IVF) of recovered blastocysts. In the group D5, the conventional IVF, better embryo development dynamics was observed, with 66% of expanded blastocysts, against 28% in the IVC. Group D3 showed 75% (IVC) and 53% (IVF) of embryo formation. Embryonic quality in Group D3 demonstrated that IVF embryos had a better synchrony in the number and quality of blastomeres. All embryos recovered in Group D3, in both techniques, did not show fragmentation. The pH of the medium contained in the INVOCell™ device in both Groups D5 and D3 showed no differences. The means were 7.26 and 7.25, respectively. The pH of the medium used in IVF was 7.29 in both groups. Microbiological analyzes of the culture media contained in the INVOCell™ devices used in Group D5 were negative. CONCLUSIONS: The results showed that the IVC technique, using the INVOCell™ device, provided a healthy and balanced environment for the development and obtaining of quality embryos with three and five days of culture.

A novel and simple heat-based method eliminates the highly detrimental effect of xylene deparaffinization on acid-fast stains.

OBJECTIVES: Histopathology is an important method for diagnosing extrapulmonary tuberculosis, yet tissue sections are often negative for mycobacteria after use of acid-fast stain (AFS). This study investigated the mechanism of AFS use and the detrimental effect of histologic processing-in particular, xylene deparaffinization-on AFS and mycobacterial detection. METHODS: The target of the fluorescent Auramine O (AuO) AFS was investigated using triple staining with DNA- and RNA-specific dyes. The effect of xylene deparaffinization on the acid fastness of mycobacteria in cultures or tissue sections was studied using AuO fluorescence as a quantitative marker. The xylene method was compared with a novel, solvent-free projected-hot-air deparaffinization (PHAD). RESULTS: Co-localization of AuO with DNA/RNA stains suggests that intracellular nucleic acids are the true target of AFS, producing highly specific patterns. Xylene reduces mycobacterial fluorescence significantly (P < .0001; moderate effect size, r = 0.33). The PHAD process yielded significantly higher fluorescence than xylene deparaffinization in tissues (P < .0001; large effect size, r = 0.85). CONCLUSIONS: Auramine O can be applied for nucleic acid staining of mycobacteria in tissues producing typical beaded patterns. Acid-fast staining depends heavily on the integrity of the mycobacterial cell wall, which xylene appears to damage. A solvent-free tissue deparaffinization method has the potential to increase mycobacterial detection significantly.

Yerba mate consumption by ovariectomized rats alters white adipose tissue.

INTRODUCTION: Menopause and post-menopause are characterized by low levels of estrogen that can be associated with the emergence of metabolic diseases. While hormone replacement therapy can alleviate many symptoms, it can also exacerbate other diseases such as breast cancer. In the search for natural alternatives, Ilex paraguariensis (Yerba Mate) has been identified as a potential therapy for the onset of obesity. Here, the effect of MATE consumption on white adipose tissue (WAT) was studied in ovariectomized rats, an animal model for post-menopause hormone loss. METHODS: Four groups of animals were used: ovariectomy with MATE (OVX MATE) and without MATE (OVX), as well as sham surgery with MATE (Sham MATE) and without MATE (Sham). MATE was provided by gavage at 1 g/kg of body weight for eight weeks before measuring biochemical parameters in plasma and characterizing WAT morphology. RESULTS: The consumption of Yerba MATE significantly decreased weight gain in ovariectomized rats and presented near control levels of triglycerides, total cholesterol, and LDL. A morphometric analysis of WAT showed a significant decrease in the area occupied by adipocytes in the group that consumed MATE. Finally, MATE consumption increased the UCP1 content in the WAT of the ovariectomized group. Yerba MATE treatment was also associated with higher levels of SIRT1 protein. CONCLUSION: MATE consumption has a preventive effect on the weight gain observed in ovariectomized rats and potential benefits in naturally avoiding the onset of obesity post menopause.

Maternal γδ T cells shape offspring pulmonary type 2 immunity in a microbiota-dependent manner.

Immune development is profoundly influenced by vertically transferred cues. However, little is known about how maternal innate-like lymphocytes regulate offspring immunity. Here, we show that mice born from γδ T cell-deficient (TCRδ-/-) dams display an increase in first-breath-induced inflammation, with a pulmonary milieu selectively enriched in type 2 cytokines and type 2-polarized immune cells, when compared with the progeny of γδ T cell-sufficient dams. Upon helminth infection, mice born from TCRδ-/- dams sustain an increased type 2 inflammatory response. This is independent of the genotype of the pups. Instead, the offspring of TCRδ-/- dams harbors a distinct intestinal microbiota, acquired during birth and fostering, and decreased levels of intestinal short-chain fatty acids (SCFAs), such as pentanoate and hexanoate. Importantly, exogenous SCFA supplementation inhibits type 2 innate lymphoid cell function and suppresses first-breath- and infection-induced inflammation. Taken together, our findings unravel a maternal γδ T cell-microbiota-SCFA axis regulating neonatal lung immunity.

Interplay between liver and blood stages of Plasmodium infection dictates malaria severity via γδ T cells and IL-17-promoted stress erythropoiesis.

Plasmodium replicates within the liver prior to reaching the bloodstream and infecting red blood cells. Because clinical manifestations of malaria only arise during the blood stage of infection, a perception exists that liver infection does not impact disease pathology. By developing a murine model where the liver and blood stages of infection are uncoupled, we showed that the integration of signals from both stages dictated mortality outcomes. This dichotomy relied on liver stage-dependent activation of Vγ4+ γδ T cells. Subsequent blood stage parasite loads dictated their cytokine profiles, where low parasite loads preferentially expanded IL-17-producing γδ T cells. IL-17 drove extra-medullary erythropoiesis and concomitant reticulocytosis, which protected mice from lethal experimental cerebral malaria (ECM). Adoptive transfer of erythroid precursors could rescue mice from ECM. Modeling of γδ T cell dynamics suggests that this protective mechanism may be key for the establishment of naturally acquired malaria immunity among frequently exposed individuals.