Acute Chagas Disease Outbreak among Military Personnel, Colombia, 2021.

We report an acute Chagas disease outbreak among soldiers in Colombia. Trypanosoma cruzi infection was confirmed through parasitology, serology, and molecular methods. Among 9 affected soldiers, 2 died; 7 were hospitalized and received benznidazole treatment, which produced favorable outcomes. Personnel patrolling rural areas in Colombia could be at increased risk for Chagas disease.

Molecular Evidence for Olfactory Neuroblastoma as a Tumor of Malignant Globose Basal Cells.

Olfactory neuroblastoma (ONB, esthesioneuroblastoma) is a sinonasal cancer with an underdeveloped diagnostic toolkit, and is the subject of many incidents of tumor misclassification throughout the literature. Despite its name, connections between the cancer and normal cells of the olfactory epithelium have not been systematically explored and markers of olfactory epithelial cell types are not deployed in clinical practice. Here, we utilize an integrated human-mouse single-cell atlas of the nasal mucosa, including the olfactory epithelium, to identify transcriptomic programs that link ONB to a specific population of stem/progenitor cells known as olfactory epithelial globose basal cells (GBCs). Expression of a GBC transcription factor NEUROD1 distinguishes both low- and high-grade ONB from sinonasal undifferentiated carcinoma, a potential histologic mimic with a distinctly unfavorable prognosis. Furthermore, we identify a reproducible subpopulation of highly proliferative ONB cells expressing the GBC stemness marker EZH2, suggesting that EZH2 inhibition may play a role in the targeted treatment of ONB. Finally, we study the cellular states comprising ONB parenchyma using single-cell transcriptomics and identify evidence of a conserved GBC transcriptional regulatory circuit that governs divergent neuronal-versus-sustentacular differentiation. These results link ONB to a specific cell type for the first time and identify conserved developmental pathways within ONB that inform diagnostic, prognostic, and mechanistic investigation.

Molecular analysis of acute and chronic reactive astrocytes in the pilocarpine model of temporal lobe epilepsy.

Astroglia, the most abundant glial cells in the mammalian central nervous system (CNS), are considered an emerging key player in seizure induction and progression. Although astrocytes undergo reactive gliosis in temporal lobe epilepsy (TLE) with dramatic morphological and molecular changes, specific astrocyte targets/molecular pathways that contribute to the induction and progression of seizure remain largely unknown. By combining translating ribosomal affinity purification (TRAP) with the pilocarpine model of TLE in BAC aldh1l1 TRAP mice, we profiled translating mRNAs from hippocampal or cortical astrocytes at different phases (3days, 30days, and 60days post-pilocarpine injections) of pilocarpine-induced epilepsy models. Our results found that hippocampal (but not cortical) astrocytes undergo early and unique molecular changes at 3days post-pilocarpine injections. These changes indicate a potentially primary pathogenic role of hippocampal astrocytes in seizure induction and progression and provide new insights about the involvement of specific astrocytic pathways/targets in epilepsy. In particular, we validated expression changes of ocrl and aeg1 in pilocarpine models. Follow-up studies on these genes may reveal new roles of hippocampal astrocytes in TLE.

Phenotypic instability between the near isogenic substrains BALB/cJ and BALB/cByJ.

Closely related substrains of inbred mice often show phenotypic differences that are presumed to be caused by recent mutations. The substrains BALB/cJ and BALB/cByJ, which were separated in 1935, have been reported to show numerous highly significant behavioral and morphological differences. In an effort to identify some of the causal mutations, we phenotyped BALB/cJ and BALB/cByJ mice as well as their F1, F2, and N2 progeny for behavioral and morphological phenotypes. We also generated whole-genome sequence data for both inbred strains (~3.5× coverage) with the intention of identifying polymorphic markers to be used for linkage analysis. We observed significant differences in body weight, the weight of the heart, liver, spleen and brain, and corpus callosum length between the two substrains. We also observed that BALB/cJ animals showed greater anxiety-like behavior in the open field test, less depression-like behavior in the tail suspension test, and reduced aggression compared to BALB/cByJ mice. Some but not all of these physiological and behavioral results were inconsistent with prior publications. These inconsistencies led us to suspect that the differences were due to, or modified by, non-genetic factors. Thus, we did not perform linkage analysis. We provide a comprehensive summary of the prior literature about phenotypic differences between these substrains as well as our current findings. We conclude that many differences between these strains are unstable and therefore ill-suited to linkage analysis; the source of this instability is unclear. We discuss the broader implications of these observations for the design of future studies.

Spatiotemporal dynamics exhibited by horizontal basal cells reveal a pro-neurogenic pathway during injury-induced olfactory epithelium regeneration.

Horizontal basal cells (HBCs) mediate olfactory epithelium (OE) regeneration following severe tissue injury. The dynamism of the post-injury environment is well illustrated by in silico modeling of RNA sequencing data that demonstrate an evolving HBC transcriptome. Unfortunately, spatiotemporally dynamic processes occurring within HBCs in situ remain poorly understood. Here, we show that HBCs at 24 h post-OE injury spatially redistribute a constellation of proteins, which, in turn, helped to nominate Rac1 as a regulator of HBC differentiation during OE regeneration. Using our primary culture model to activate HBCs pharmacologically, we demonstrate that concurrent Rac1 inhibition attenuates HBC differentiation potential. This in vitro functional impairment manifested in vivo as decreased HBC differentiation into olfactory sensory neurons following HBC-specific Rac1 conditional knockout. Taken together, our data potentiate the design of hyposmia-alleviating therapies and highlight aspects of in situ HBC spatiotemporal dynamics that deserve further investigation.

Cardiac dysfunction in sucrose-fed rats is associated with alterations of phospholamban phosphorylation and TNF-α levels.

INTRODUCTION: High sucrose intake is linked to cardiovascular disease, a major global cause of mortality worldwide. Calcium mishandling and inflammation play crucial roles in cardiac disease pathophysiology. OBJECTIVE: Evaluate if sucrose-induced obesity is related to deterioration of myocardial function due to alterations in the calcium-handling proteins in association with proinflammatory cytokines. METHODS: Wistar rats were divided into control and sucrose groups. Over eight weeks, Sucrose group received 30% sucrose water. Cardiac function was determined in vivo using echocardiography and in vitro using papillary muscle assay. Western blotting was used to detect calcium handling protein; ELISA assay was used to assess TNF-α and IL-6 levels. RESULTS: Sucrose led to cardiac dysfunction. RYR2, SERCA2, NCX, pPBL Ser16 and L-type calcium channels were unchanged. However, pPBL-Thr17, and TNF-α levels were elevated in the S group. CONCLUSION: Sucrose induced cardiac dysfunction and decreased myocardial contractility in association with altered pPBL-Thr17 and elevated cardiac pro-inflammatory TNF-α.

An in vitro model of acute horizontal basal cell activation reveals dynamic gene regulatory networks underlying the acute activation phase.

Horizontal basal cells (HBCs) activate only in response to severe olfactory epithelium (OE) injury. This activation is mediated by the loss of the transcription factor TP63. Using the compound phorbol 12-myristate 13-acetate (PMA), we find that we can model the process of acute HBC activation. First, we find that PMA treatment induces a rapid loss in TP63 protein and induces the expression of HOPX and the nuclear translocation of RELA, previously identified to mediate HBC activation. Using bulk RNA sequencing, we find that PMA-treated HBCs pass through various stages of acute activation identifiable by transcriptional regulatory signatures that mimic stages identified in vivo . These temporal stages are associated with varying degrees of engraftment and differentiation potential in transplantation assays. Together, this data shows that our model can model physiologically relevant features of HBC activation and identifies new candidates for mechanistic testing.

Effects of glyphosate exposure on western diet-induced non-alcoholic fatty liver disease in mice.

We evaluated whether glyphosate promotes western diet (WD)-induced non-alcoholic fatty liver disease (NAFLD). Male C57BL/6J mice were fed WD and received intragastrical glyphosate (0.05, 5 or 50 mg/kg) for 6 months. Glyphosate did not promote WD-induced obesity, hypercholesterolemia, glucose intolerance, hepatic steatosis, and fibrosis. Nonetheless, the higher dose (50 mg) enhanced hepatic CD68+ macrophage density, p65, TNF-α, and IL-6 protein levels. Furthermore, this dose decreased hepatic Nrf2 levels, while enhancing lipid peroxidation in the liver and adipose tissue. Hepatic transcriptome revealed that glyphosate at 50 mg upregulated 212 genes and downregulated 731 genes. Genes associated with oxidative stress and inflammation were upregulated, while key cell cycle-related genes were downregulated. Our results indicate that glyphosate exposure - in a dose within the toxicological limits - impairs hepatic inflammation/redox dynamics in a NAFLD microenvironment.

Activating a Reserve Neural Stem Cell Population In Vitro Enables Engraftment and Multipotency after Transplantation.

The olfactory epithelium (OE) regenerates after injury via two types of tissue stem cells: active globose cells (GBCs) and dormant horizontal basal cells (HBCs). HBCs are roused to activated status by OE injury when P63 levels fall. However, an in-depth understanding of activation requires a system for culturing them that maintains both their self-renewal and multipotency while preventing spontaneous differentiation. Here, we demonstrate that mouse, rat, and human HBCs can be cultured and passaged as P63+ multipotent cells. HBCs in vitro closely resemble HBCs in vivo based on immunocytochemical and transcriptomic comparisons. Genetic lineage analysis demonstrates that HBCs in culture arise from both tissue-derived HBCs and multipotent GBCs. Treatment with retinoic acid induces neuronal and non-neuronal differentiation and primes cultured HBCs for transplantation into the lesioned OE. Engrafted HBCs generate all OE cell types, including olfactory sensory neurons, confirming that HBC multipotency and neurocompetency are maintained in culture.

Identification of clinically relevant phenotypes in patients with Ebstein anomaly.

BACKGROUND: Ebstein anomaly (EA) is a heterogeneous congenital heart defect (CHD), frequently accompanied by diverse cardiac and extracardiac comorbidities, resulting in a wide range of clinical outcomes. HYPOTHESIS: Phenotypic characterization of EA patients has the potential to identify variables that influence prognosis and subgroups with distinct contributing factors. METHODS: A comprehensive cross-sectional phenotypic characterization of 147 EA patients from one of the main referral institutions for CHD in Colombia was carried out. The most prevalent comorbidities and distinct subgroups within the patient cohort were identified through cluster analysis. RESULTS: The most prevalent cardiac comorbidities identified were atrial septal defect (61%), Wolff-Parkinson-White syndrome (WPW; 27%), and right ventricular outflow tract obstruction (25%). Cluster analysis showed that patients can be classified into 2 distinct subgroups with defined phenotypes that determine disease severity and survival. Patients in cluster 1 represented a particularly homogeneous subgroup with a milder spectrum of disease, including only patients with WPW and/or supraventricular tachycardia (SVT). Cluster 2 included patients with more diverse cardiovascular comorbidities. CONCLUSIONS: This study represents one of the largest phenotypic characterizations of EA patients reported. The data show that EA is a heterogeneous disease, very frequently associated with cardiovascular and noncardiovascular comorbidities. Patients with WPW and SVT represent a homogeneous subgroup that presents with a less severe spectrum of disease and better survival when adequately managed. This should be considered when searching for genetic causes of EA and in the clinical setting.

Genetic Background Limits Generalizability of Genotype-Phenotype Relationships.

Genome-wide association studies (GWASs) have identified numerous loci that influence risk for psychiatric diseases. Genetically engineered mice are often used to characterize genes implicated by GWASs. These studies are based on the assumption that observed genotype-phenotype relationships will generalize to humans, implying that the results would at least generalize to other inbred mouse strains. Given current concerns about reproducibility, we sought to directly test this assumption. We produced F1 crosses between male C57BL/6J mice heterozygous for null alleles of Cacna1c and Tcf7l2 and wild-type females from 30 inbred laboratory strains. We found extremely strong interactions with genetic background that sometimes supported diametrically opposing conclusions. These results do not negate the invaluable contributions of mouse genetics to biomedical science, but they do show that genotype-phenotype relationships cannot be reliably inferred by studying a single genetic background, and thus constitute a major challenge to the status quo. VIDEO ABSTRACT.

Série de casos de neoplasia intraepitelial vulvar tratados com imiquimode / Case series of vulvar intraepithelial neoplasia treated with imiquimod

O termo neoplasia intraepitelial vulvar (NIV) inclui as lesões precursoras do câncer de vulva, sendo divididas em NIV escamosa e não escamosa. A NIV escamosa compreende a NIV tipo usual, a qual se encontra relacionada à infecção pelo HPV, à multicentricidade de lesões, à atividade sexual e ao fumo,sendo mais frequentemente observada em mulheres jovens. Já a NIV diferenciada está relacionada ao líquen escleroso, não estando associada à infecção induzida por HPV, nem ao comportamento sexual, acometendo pacientes mais idosas. A NIV não escamosa está relacionada à doença de Paget e ao melanoma in situ. O tratamento padrão da NIV é cirúrgico e consiste na excisão ampla da lesão com margem de segurança. Este estudo objetiva avaliar ouso do imiquimode creme a 5% no tratamento da NIV tipo usual de forma isolada e/ou, associado ao procedimento cirúrgico. O imiquimode já é utilizadocom eficácia no tratamento de lesões condilomatosas genitais. Além disso, tem-se mostrado eficaz em lesões de NIV tipo usual.

The term vulvar intraepithelial neoplasia (VIN) includes the precursor lesions of vulvar cancer, being divided into squamous VIN and non-squamous VIN. The squamous VIN refers to the usual type VIN, which is related to HPV infection, multicentricity of lesions, sexual activity and smoking beingmore frequently observed in young woman. The VIN differentiated type, which is related to vulvar dermatoses, such as lichen sclerosus, is not associated with HPV-induced infection or sexual behavior and affects older patients. Thenon-squamous VIN is related to Paget's disease and melanoma in "situ".The standard VIN's treatment is a surgical wide excision of the lesion with a safety margin. This study aims to test the use of imiquimod 5% cream in thetreatment of usual type VIN isolated, and/or associated with surgical procedure. The imiquimod is already successfully used in the treatment of genital condylomatous lesions. Moreover, it has been shown to be effective in usual type VIN lesions.

Aterosclerose, uma resposta inflamatória / Atherosclerosis, an inflammatory response

A aterosclerose é a doença responsável pelo maior índice de morbidade e mortalidade no mundo. A lesão aterosclerótica é a anormalidade mais comum encontrada nas artérias, decorrente inicialmente de dois processos básicos: acúmulo de colesterol e proliferação de células musculares lisas na túnica íntima o que leva à inflamação. Esse processo dará origem a uma placa fibrosa que se projeta para dentro do lúmen, modificando a túnica média, levando a uma série de complicações circulatórias decorrentes da resposta inflamatória desencadeada na parede do vaso. Assim, nesta revisão veremos o envolvimento da resposta inflamatória e do estresse oxidativo no desencadeamento e no estabelecimento da doença aterosclerótica promovido por macrófagos, uma das principais células envolvidas nesse processo, além de discutir sobre os principais marcadores bioquímicos como as citocinas, proteínas de fase aguda e moléculas de adesão.

Atherosclerosis and the direct outcomes of ischemia are the major causes of morbidity and mortality worldwide.Dysfunction of the vascular endothelium can produce atherosclerotic disease processes with consequentinflammation, which is a significant component of atherosclerosis lesions. Atherosclerosis is characterizedby chronic inflammation and enrichment of inflammatory cells in the vessel wall. This review besides focusingon both the inflammatory response and the oxidative stress that play a major role in the atherogenesis and inthe development of cardiovascular disease stimulated by macrophages, the key cell involved in this process,will in addition discuss the several biochemical markers such as cytokines, acute phase proteins, and cellularadhesion molecules.

Estudo da ativação de monócitos e produção de citocinas em pacientes com aterosclerose obliterante periférica / Study of the activation of monocytes and peripheral production of cytokines in patients with atherosclerosis obleterans

O presente trabalho teve como objetivos avaliar em pacientes com aterosclerose obliterante periférica os níveis plasmáticos das citocinas pró e anti-inflamatórias fator de necrose tumoral-alfa (TNF-alfa) interleucina 6 (IL-6), interferon – gama (IFN-gama), interleucina 10 (IL-10) e fator transformador de crescimento beta (TGF- beta), assim como o grau de ativação de monócitos do sangue periférico, através da capacidade dessas células liberarem peróxido de hidrogênio (H2O2) e desenvolverem atividade fungicida contra Cândida albicans (C. albicans). Foram avaliados 8 indivíduos do sexo masculino, acima de 60 anos portadores de claudicação intermitente moderada e 8 indivíduos do sexo masculino acima de 60 anos que não apresentam doença arterial periférica. Observamos que os níveis plasmáticos de todas as citocinas testadas foram significativamente mais elevados nos pacientes, quando comparados aos detectados nos indivíduos controle. A capacidade das células de indivíduos controle e pacientes de liberarem H2O2 foi avaliada antes e após a incubação com citocinas envolvidas no processo de ativação dessas células, como o IFN-gama e TNF- alfa. Observamos que monócitos de indivíduos controle não ativados liberaram níveis substanciais do metabólito que, no entanto, aumentaram significativamente após a ativação com IFN-gama ou TNF-alfa. As células dos pacientes apresentaram um perfil de resposta semelhante aos apresentados pelos indivíduos controles. No entanto, quando comparamos a resposta dos dois grupos detectamos que os monócitos dos pacientes antes e após o processo de ativação, apresentam respostas menores que os indivíduos controle. Resultados semelhantes foram detectados nos ensaios de avaliação da atividade fungicida isto é, células de controles e pacientes apresentaram um aumento da atividade fungicida após a ativação tanto com IFN-gama como com TNF-alfa. No entanto, os resultados dos pacientes foram sempre significativamente menores que os detectados para con...