Amphotericin B resistance in Leishmania amazonensis: In vitro and in vivo characterization of a Brazilian clinical isolate.

In Brazil, Leishmania amazonensis is the etiological agent of cutaneous and diffuse cutaneous leishmaniasis. The state of Maranhão in the Northeast of Brazil is prevalent for these clinical forms of the disease and also has high rates of HIV infection. Here, we characterized the drug susceptibility of a L. amazonensis clinical isolate from a 46-year-old man with diffuse cutaneous leishmaniasis coinfected with HIV from this endemic area. This patient underwent several therapeutic regimens with meglumine antimoniate, liposomal amphotericin B, and pentamidine, without success. In vitro susceptibility assays against promastigotes and intracellular amastigotes demonstrated that this isolate had low susceptibility to amphotericin B, when compared with the reference strain of this species that is considered susceptible to antileishmanial drugs. Additionally, we investigated whether the low in vitro susceptibility would affect the in vivo response to amphotericin B treatment. The drug was effective in reducing the lesion size and parasite burden in mice infected with the reference strain, whereas those infected with the clinical isolate and a resistant line (generated experimentally by stepwise selection) were refractory to amphotericin B treatment. To evaluate whether the isolate was intrinsically resistant to amphotericin B in animals, infected mice were treated with other drugs that had not been used in the treatment of the patient (miltefosine, paromomycin, and a combination of both). Our findings demonstrated that all drug schemes were able to reduce lesion size and parasite burden in animals infected with the clinical isolate, confirming the amphotericin B-resistance phenotype. These findings indicate that the treatment failure observed in the patient may be associated with amphotericin B resistance, and demonstrate the potential emergence of amphotericin B-resistant L. amazonensis isolates in an area of Brazil endemic for cutaneous leishmaniasis.

The molecular basis of phenotypic evolution: beyond the usual suspects.

It has been well documented that mutations in coding DNA or cis-regulatory elements underlie natural phenotypic variation in many organisms. However, the development of sophisticated functional tools in recent years in a wide range of traditionally non-model systems have revealed many 'unusual suspects' in the molecular bases of phenotypic evolution, including upstream open reading frames (uORFs), cryptic splice sites, and small RNAs. Furthermore, large-scale genome sequencing, especially long-read sequencing, has identified a cornucopia of structural variation underlying phenotypic divergence and elucidated the composition of supergenes that control complex multi-trait polymorphisms. In this review article we highlight recent studies that demonstrate this great diversity of molecular mechanisms producing adaptive genetic variation and the panoply of evolutionary paths leading to the 'grandeur of life'.

Understanding COVID-19 progression with longitudinal peripheral blood mononuclear cell proteomics: Changes in the cellular proteome over time.

The clinical presentation of COVID-19 is highly variable, and understanding the underlying biological processes is crucial. This study utilized a proteomic analysis to investigate dysregulated processes in the peripheral blood mononuclear cells of patients with COVID-19 compared to healthy volunteers. Samples were collected at different stages of the disease, including hospital admission, after 7 days of hospitalization, and 30 days after discharge. Metabolic pathway alterations and increased abundance of neutrophil-related proteins were observed in patients. Patients progressing to critical illness had significantly low-abundance proteins in the pentose phosphate and glycolysis pathways compared with those presenting clinical recovery. Important biological processes, such as fatty acid concentration and glucose metabolism disorder, remained altered even after 30 days of hospital discharge. Temporal proteomic changes revealed distinct pathways in critically ill and non-critically ill patients. Our study emphasizes the significance of longitudinal cellular proteomic studies in identifying disease progression-related pathways and persistent protein changes post-hospitalization.

In Vitro Drug Susceptibility of a Leishmania (Leishmania) infantum Isolate from a Visceral Leishmaniasis Pediatric Patient after Multiple Relapses.

The parasitic protozoan Leishmania (Leishmania) infantum is the etiological agent of human visceral leishmaniasis in South America, an infectious disease associated with malnutrition, anemia, and hepatosplenomegaly. In Brazil alone, around 2700 cases are reported each year. Treatment failure can occur as a result of drug, host, and/or parasite-related factors. Here, we isolated a Leishmania species from a pediatric patient with visceral leishmaniasis that did not respond to chemotherapy, experiencing a total of nine therapeutic relapses and undergoing a splenectomy. The parasite was confirmed as L. (L.) infantum after sequencing of the ribosomal DNA internal transcribed spacer, and the clinical isolate, in both promastigote and amastigote forms, was submitted to in vitro susceptibility assays with all the drugs currently used in the chemotherapy of leishmaniasis. The isolate was susceptible to meglumine antimoniate, amphotericin B, pentamidine, miltefosine, and paromomycin, similarly to another strain of this species that had previously been characterized. These findings indicate that the multiples relapses observed in this pediatric patient were not due to a decrease in the drug susceptibility of this isolate; therefore, immunophysiological aspects of the patient should be further investigated to understand the basis of treatment failure in this case.

Expression of TIGIT, PD-1 and HLA-DR/CD38 markers on CD8-T cells of children and adolescents infected with HIV and uninfected controls.

Immune exhaustion and senescence are scarcely studied in HIV-pediatric patients. We studied the circulatory CD8 T cells activation/exhaustion and senescent phenotype of children and adolescents vertically infected with HIV or uninfected controls based on the expression of human leukocyte antigen (HLA-DR), CD38, T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif (ITIM) domain (TIGIT), programmed death 1 (PD-1) and CD57 by flow cytometry, during approximately one year. Eleven HIV-infected (HI) and nine HIV-uninfected (HU) children/adolescents who received two doses or one dose of meningococcal C conjugate vaccine (MenC), respectively, were involved in this study. Blood samples were collected before the immunization (T0), 1-2 months after the first dose (T1), and 1-2 months after the second dose (T2), which was administered approximately one year after the first one. HI patients not receiving combined antiretroviral therapy (cART) showed a higher frequency of CD8 T cells TIGIT+, PD-1+ or CD57+, as well as a higher frequency of CD8 T cells co-expressing CD38/HLA-DR/TIGIT or CD38/HLA-DR/PD-1 when compared to HI treated or HU individuals, at all times that they were assessed. CD8 T cells co-expressing CD38/DR/TIGIT were inversely correlated with the CD4/CD8 ratio but positively associated with viral load. The co-expression of CD38/DR/TIGIT or CD38/DR/PD-1 on CD8 T cells was also inversely associated with the CD4 T cells expressing co-stimulatory molecules CD127/CD28. The results showed a higher expression of exhaustion/senescence markers on CD8 T cells of untreated HI children/adolescents and its correlations with viral load.

In vitro miltefosine and amphotericin B susceptibility of strains and clinical isolates of Leishmania species endemic in Brazil that cause tegumentary leishmaniasis.

Tegumentary leishmaniasis encompasses a spectrum of clinical manifestations caused by the parasitic protozoa of the genus Leishmania. In Brazil, there are at least seven Leishmania species that are endemic and responsible for this set of clinical manifestations of the disease. Current treatment is limited to a restricted number of drugs that in general have several drawbacks including parenteral use, toxicity, and severe side effects. Amphotericin B is considered a second-line drug for tegumentary leishmaniasis in Brazil, while miltefosine was recently approved for clinical use in the treatment of this disease. In this study, we investigated the in vitro susceptibility of Leishmania strains representative of the species endemic to Brazil, as well as a panel of thirteen clinical isolates of tegumentary leishmaniasis, to both amphotericin B and miltefosine. A moderate variation in the susceptibility to both drugs was found, where the EC50 values varied from 11.43 to 52.67 µM for miltefosine and from 12.89 to 62.36 nM for amphotericin B in promastigotes, while for the intracellular amastigotes, values ranged from 1.08 to 9.60 µM and from 1.69 to 22.71 nM for miltefosine and amphotericin B respectively. Furthermore, the clinical isolates and strains of the subgenus Viannia were evaluated for the presence of Leishmania RNA virus 1 (LRV1), as this is an important factor associated with disease severity and treatment outcome. These findings provide a preclinical dataset of the activity of these drugs against the causative species of tegumentary leishmaniasis in Brazil.

Práticas de higiene bucal em bebês de 6 meses de idade / Oral hygiene practices in babies aged 6 months

Resumo: Introdução a recomendação mais atual é de que a higiene bucal deve ser iniciada após a erupção do primeiro dente. Porém, há divergência de opiniões a respeito da necessidade ou não da higiene bucal do bebê desdentado. Objetivo: avaliar as práticas maternas de higienização bucal dos bebês menores de 6 meses de idade. Material e método : foi aplicado um formulário digital contendo quatro questões sobre informações sociodemográficas e quatro questões específicas. Foi realizada análise estatística descritiva, e os resultados foram comparados estatisticamente entre as crianças que receberam e não receberam higiene bucal antes dos 6 meses de idade. Resultado: pode-se observar que a idade, a escolaridade e a renda familiar das mães que realizavam higiene bucal de seus filhos antes da erupção do primeiro dente foram maiores do que das mães que não realizavam esse tipo de higiene. A idade das crianças, a idade de erupção do primeiro dente, o recebimento de orientações profissionais e o tipo de aleitamento não diferiram entre os grupos. Conclusão: ainda há uma grande parcela de mães que realiza higiene bucal dos bebês edêntulos.

Abstract: Introduction the starts of oral hygiene after the first tooth eruption is the most current recommendation. However, there are differences of opinion regarding whether or not oral hygiene is necessary for edentulous babies. Objective: to evaluate the maternal practices of oral hygiene of babies under 6 months of age. Material and method: a digital form was applied containing 4 questions about sociodemographic information and 4 specific questions. Descriptive statistical analysis was performed and the results were statistically compared between children who received and did not receive oral hygiene before 6 months of age. Result: it can be observed that the age, education and family income of mothers who performed oral hygiene before the eruption of the first tooth was higher than that of mothers who did not perform this type of hygiene. The age of the children, age of eruption of the first tooth, whether they received professional guidance and type of breastfeeding did not differ between the groups. Conclusion: there is still a large portion of mothers who perform oral hygiene of edentulous babies.

Expression of TIGIT, PD-1 and HLA-DR/CD38 markers on CD8-T cells of children and adolescents infected with HIV and uninfected controls

ABSTRACT Immune exhaustion and senescence are scarcely studied in HIV-pediatric patients. We studied the circulatory CD8 T cells activation/exhaustion and senescent phenotype of children and adolescents vertically infected with HIV or uninfected controls based on the expression of human leukocyte antigen (HLA-DR), CD38, T cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif (ITIM) domain (TIGIT), programmed death 1 (PD-1) and CD57 by flow cytometry, during approximately one year. Eleven HIV-infected (HI) and nine HIV-uninfected (HU) children/adolescents who received two doses or one dose of meningococcal C conjugate vaccine (MenC), respectively, were involved in this study. Blood samples were collected before the immunization (T0), 1-2 months after the first dose (T1), and 1-2 months after the second dose (T2), which was administered approximately one year after the first one. HI patients not receiving combined antiretroviral therapy (cART) showed a higher frequency of CD8 T cells TIGIT+, PD-1+ or CD57+, as well as a higher frequency of CD8 T cells co-expressing CD38/HLA-DR/TIGIT or CD38/HLA-DR/PD-1 when compared to HI treated or HU individuals, at all times that they were assessed. CD8 T cells co-expressing CD38/DR/TIGIT were inversely correlated with the CD4/CD8 ratio but positively associated with viral load. The co-expression of CD38/DR/TIGIT or CD38/DR/PD-1 on CD8 T cells was also inversely associated with the CD4 T cells expressing co-stimulatory molecules CD127/CD28. The results showed a higher expression of exhaustion/senescence markers on CD8 T cells of untreated HI children/adolescents and its correlations with viral load.

Construction of face databases for tasks to recognize facial expressions of basic emotions: a systematic review.

Recognizing the other's emotions is an important skill for the social context that can be modulated by variables such as gender, age, and race. A number of studies seek to elaborate specific face databases to assess the recognition of basic emotions in different contexts. Objectives: This systematic review sought to gather these studies, describing and comparing the methodologies used in their elaboration. Methods: The databases used to select the articles were the following: PubMed, Web of Science, PsycInfo, and Scopus. The following word crossing was used: "Facial expression database OR Stimulus set AND development OR Validation." Results: A total of 36 articles showed that most of the studies used actors to express the emotions that were elicited from specific situations to generate the most spontaneous emotion possible. The databases were mainly composed of colorful and static stimuli. In addition, most of the studies sought to establish and describe patterns to record the stimuli, such as color of the garments used and background. The psychometric properties of the databases are also described. Conclusions: The data presented in this review point to the methodological heterogeneity among the studies. Nevertheless, we describe their patterns, contributing to the planning of new research studies that seek to create databases for new contexts.

Reconhecer as emoções do outro é uma habilidade importante para o contexto social, que pode ser modulada por variáveis como sexo, idade e raça. Vários estudos buscam elaborar bancos de faces específicos para avaliar o reconhecimento de emoções básicas em diferentes contextos. Objetivos: Esta revisão sistemática buscou reunir esses estudos, descrevendo e comparando as metodologias utilizadas em sua elaboração. Métodos: As bases de dados utilizadas para a seleção dos artigos foram: PubMed, Web of Science, PsycInfo e Scopus. Foi utilizado o seguinte cruzamento de palavras: "facial expression database OR stimulus set AND development OR validation". Resultados: O total de 36 artigos mostrou que a maioria dos estudos utilizou atores para expressar as emoções, que foram suscitadas de situações específicas para serem o mais espontâneas possível. Os bancos de faces foram compostos principalmente de estímulos coloridos e estáticos. Além disso, a maioria dos estudos buscou estabelecer e descrever padrões para registrar os estímulos, como a cor das roupas utilizadas e o fundo. As propriedades psicométricas dos bancos de faces também são descritas. Conclusões: Os dados apresentados nesta revisão apontam para a heterogeneidade metodológica entre os estudos. Apesar disso, descrevemos seus padrões, contribuindo para o planejamento de novas pesquisas que buscam criar bancos de faces específicos para novos contextos.

Glucose metabolism is upregulated in the mononuclear cell proteome during sepsis and supports endotoxin-tolerant cell function.

Metabolic adaptations shape immune cell function. In the acute response, a metabolic switch towards glycolysis is necessary for mounting a proinflammatory response. During the clinical course of sepsis, both suppression and activation of immune responses take place simultaneously. Leukocytes from septic patients present inhibition of cytokine production while other functions such as phagocytosis and production of reactive oxygen species (ROS) are preserved, similarly to the in vitro endotoxin tolerance model, where a first stimulation with lipopolysaccharide (LPS) affects the response to a second stimulus. Here, we sought to investigate how cellular metabolism is related to the modulation of immune responses in sepsis and endotoxin tolerance. Proteomic analysis in peripheral blood mononuclear cells (PBMCs) from septic patients obtained at intensive care unit admission showed an upregulation of proteins related to glycolysis, the pentose phosphate pathway (PPP), production of ROS and nitric oxide, and downregulation of proteins in the tricarboxylic acid cycle and oxidative phosphorylation compared to healthy volunteers. Using the endotoxin-tolerance model in PBMCs from healthy subjects, we observed increased lactate production in control cells upon LPS stimulation, while endotoxin-tolerant cells presented inhibited tumor necrosis factor-α and lactate production along with preserved phagocytic capacity. Inhibition of glycolysis and PPP led to impairment of phagocytosis and cytokine production both in control and in endotoxin-tolerant cells. These data indicate that glucose metabolism supports leukocyte functions even in a condition of endotoxin tolerance.

Construction of face databases for tasks to recognize facial expressions of basic emotions: a systematic review / Construção de bancos de faces para tarefas de reconhecimento de expressões faciais de emoções básicas: uma revisão sistemática

ABSTRACT. Recognizing the other's emotions is an important skill for the social context that can be modulated by variables such as gender, age, and race. A number of studies seek to elaborate specific face databases to assess the recognition of basic emotions in different contexts. Objectives: This systematic review sought to gather these studies, describing and comparing the methodologies used in their elaboration. Methods: The databases used to select the articles were the following: PubMed, Web of Science, PsycInfo, and Scopus. The following word crossing was used: "Facial expression database OR Stimulus set AND development OR Validation." Results: A total of 36 articles showed that most of the studies used actors to express the emotions that were elicited from specific situations to generate the most spontaneous emotion possible. The databases were mainly composed of colorful and static stimuli. In addition, most of the studies sought to establish and describe patterns to record the stimuli, such as color of the garments used and background. The psychometric properties of the databases are also described. Conclusions: The data presented in this review point to the methodological heterogeneity among the studies. Nevertheless, we describe their patterns, contributing to the planning of new research studies that seek to create databases for new contexts.

RESUMO. Reconhecer as emoções do outro é uma habilidade importante para o contexto social, que pode ser modulada por variáveis como sexo, idade e raça. Vários estudos buscam elaborar bancos de faces específicos para avaliar o reconhecimento de emoções básicas em diferentes contextos. Objetivos: Esta revisão sistemática buscou reunir esses estudos, descrevendo e comparando as metodologias utilizadas em sua elaboração. Métodos: As bases de dados utilizadas para a seleção dos artigos foram: PubMed, Web of Science, PsycInfo e Scopus. Foi utilizado o seguinte cruzamento de palavras: "facial expression database OR stimulus set AND development OR validation". Resultados: O total de 36 artigos mostrou que a maioria dos estudos utilizou atores para expressar as emoções, que foram suscitadas de situações específicas para serem o mais espontâneas possível. Os bancos de faces foram compostos principalmente de estímulos coloridos e estáticos. Além disso, a maioria dos estudos buscou estabelecer e descrever padrões para registrar os estímulos, como a cor das roupas utilizadas e o fundo. As propriedades psicométricas dos bancos de faces também são descritas. Conclusões: Os dados apresentados nesta revisão apontam para a heterogeneidade metodológica entre os estudos. Apesar disso, descrevemos seus padrões, contribuindo para o planejamento de novas pesquisas que buscam criar bancos de faces específicos para novos contextos.

Dermoscopic Findings of Discoid Lupus Erythematosus Involving the Eyelids.

Introduction: Although facial involvement in discoid lupus erythematosus (DLE) is common, eyelid involvement is atypical. Identifying this condition is challenging due to misdiagnosis, and it is essential to avoid potential deformities of the eyelid margin. Case Presentation: We, herein, report the dermoscopic findings in 2 female patients with a confirmed diagnosis of DLE who presented eyelids involvement. Discussion/Conclusion: This study highlights the importance of performing a dermoscopy examination to help physicians obtain an early diagnosis of DLE.

Isolation, typing, and drug susceptibility of Leishmania (Leishmania) infantum isolates from dogs of the municipality of Embu das Artes, an endemic region for canine leishmaniasis in Brazil.

The parasitic protozoa Leishmania (Leishmania) infantum is the etiological agent of human visceral leishmaniasis and canine leishmaniasis in South America, where Brazil is the most affected country. This zoonotic disease is transmitted by the bite of an infected phlebotomine sand fly and dogs constitute the main domestic reservoir of the parasite. In this study, we screened 2348 dogs of the municipality of Embu das Artes, Brazil, for antibodies against the parasite. Prevalence for canine leishmaniasis seropositivity was 2.81%, as assessed using a Dual-Path Platform rapid test for canine leishmaniasis. Twenty-five seropositive dogs were euthanized for parasite isolation and 14 isolates were successful obtained. Nucleotide sequencing of the internal transcribed spacer confirmed the isolates to be L. (L.) infantum, and very low sequence variability was observed among them. The in vitro susceptibility to miltefosine and paromomycin was assessed and moderate variation in paromomycin susceptibility was found among the isolates in the promastigote and intracellular amastigote stages. On the other hand, in vitro susceptibility to miltefosine of these isolates was homogenous, particularly in the amastigote stage (EC50 values from 0.69 to 2.07 µM). In addition, the miltefosine sensitivity locus was deleted in all the isolates, which does not corroborate the hypothesis that the absence of this locus is correlated with a low in vitro susceptibility. Our findings confirm that the municipality of Embu das Artes is endemic for canine leishmaniasis and that isolates from this region are susceptible to paromomycin and miltefosine, indicating the potential of these drugs to be clinically evaluated in the treatment of human visceral leishmaniasis in Brazil.

Saberes e práticas de cuidado em saúde sobre a covid-19: uma análise baseada em interações de pessoas em comunidade virtual / Knowledge and health care practices of covid-19: an analysis based on interactions of people in a virtual community / Conocimientos y prácticas de atención a la salud de la covid-19: un análisis a partir deinteracciones de personas en una comunidad virtual

Este artigo analisa os saberes e práticas do cuidado em saúde relacionados à covid-19, a partir da observação participante de pessoas que trocam experiências e interagem sobre o tema no grupo da rede social Facebook, no Brasil, 'Eu já tive Covid-19'. A comunidade virtual, com 16,5 mil membros, gerou 397 postagens no período de 8 de março a 18 de abril de 2021. As postagens foram categorizadas e tiveram seus níveis de engajamento calculados com base no número de interações. Experiências em casos de covid-19, dúvidas e questionamentos dos usuários participantes representaram 74% das postagens. Os comentários com mensagens de acolhimento aproximaram as pessoas, fortalecendo seus laços sociais. Foram identificados discursos que contrariavam o saber científico, sendo uma a cada 25 postagens definida como fake news. Temas como automedicação, xarope caseiro, 'kit covid' e tratamento precoce foram destaque dentre os conteúdos e evidenciam o grau de desinformação dos participantes a respeito da covid-19.

This article analyzes the knowledge and practices of health care related to covid-19, based on the participant observation of people who exchange experiences and interact on the topic in the social network Facebook group, in Brazil, 'I already had Covid-19'. The virtual community, with 16,500 members, generated 397 posts, in the period from March 8 to April 18, 2021. They were categorized and had their engagement levels calculated based on the number of interactions. Experiences in cases of covid-19, doubts and questions from participating users represented 74% of posts. The comments with welcoming messages brought these people closer, strengthening their social ties. Speeches that contradicted scientific knowledge were identified, with one in 25 posts classified as fake news. Topics such as self-medication, homemade syrup, 'covid kit' and early treatment were highlighted among these contents and show the degree of disinformation of the participants about covid-19.

Este artículo analiza los conocimientos y prácticas de atención a la salud relacionados con el covid-19, a partir de la observación participante de personas que intercambian experiencias e interactúan sobre el tema en el grupo de la red social Facebook, en Brasil, 'Eu ya tuvo Covid-19'. La comunidad virtual, con 16.500 miembros, generó 397 publicaciones, en el período del 8 de marzo al 18 de abril de 2021. Se categorizaron y se calcularon los niveles de participación en función del número de interacciones. Experiencias en casos de covid-19, dudas y preguntas de los usuarios participantes representaron el 74% de las publicaciones. Los comentarios con mensajes de bienvenida acercaron a estas personas, fortaleciendo sus lazos sociales. Se identificaron discursos que contradecían el conocimiento científico, con una fake news cada 25 publicaciones. En estos contenidos se destacaron temas como la automedicación, el jarabe casero, el 'kit covid' y el tratamiento temprano, evidenciando el grado de desinformación de los participantes sobre el covid-19.

The PPAR-γ agonist pioglitazone exerts proinflammatory effects in bronchial epithelial cells during acute Pseudomonas aeruginosa pneumonia.

Pseudomonas aeruginosa is a common respiratory pathogen that causes injurious airway inflammation during acute pneumonia. Peroxisome proliferator-activated receptor (PPAR)-γ is involved in the regulation of metabolic and inflammatory responses in different cell types and synthetic agonists of PPAR-γ exert anti-inflammatory effects on myeloid cells in vitro and in models of inflammation in vivo. We sought to determine the effect of the PPAR-γ agonist pioglitazone on airway inflammation induced by acute P. aeruginosa pneumonia, focusing on bronchial epithelial cells. Mice pretreated with pioglitazone or vehicle (24 and 1 h) were infected with P. aeruginosa via the airways. Pioglitazone treatment was associated with increased expression of chemokine (Cxcl1, Cxcl2, and Ccl20) and cytokine genes (Tnfa, Il6, and Cfs3) in bronchial brushes obtained 6 h after infection. This pro-inflammatory effect was accompanied by increased expression of Hk2 and Pfkfb3 genes encoding rate-limiting enzymes of glycolysis; concurrently, the expression of Sdha, important for maintaining metabolite flux in the tricarboxylic acid cycle, was reduced in bronchial epithelial cells of pioglitazone treated-mice. Pioglitazone inhibited bronchoalveolar inflammatory responses measured in lavage fluid. These results suggest that pioglitazone exerts a selective proinflammatory effect on bronchial epithelial cells during acute P. aeruginosa pneumonia, possibly by enhancing intracellular glycolysis.

HIF-1α Stabilization in Flagellin-Stimulated Human Bronchial Cells Impairs Barrier Function.

The respiratory epithelium provides a first line of defense against pathogens. Hypoxia-inducible factor (HIF)1α is a transcription factor which is stabilized in hypoxic conditions through the inhibition of prolyl-hydroxylase (PHD)2, the enzyme that marks HIF1α for degradation. Here, we studied the impact of HIF1α stabilization on the response of primary human bronchial epithelial (HBE) cells to the bacterial component, flagellin. The treatment of flagellin-stimulated HBE cells with the PHD2 inhibitor IOX2 resulted in strongly increased HIF1α expression. IOX2 enhanced the flagellin-induced expression of the genes encoding the enzymes involved in glycolysis, which was associated with the intracellular accumulation of pyruvate. An untargeted pathway analysis of RNA sequencing data demonstrated the strong inhibitory effects of IOX2 toward key innate immune pathways related to cytokine and mitogen-activated kinase signaling cascades in flagellin-stimulated HBE cells. Likewise, the cell-cell junction organization pathway was amongst the top pathways downregulated by IOX2 in flagellin-stimulated HBE cells, which included the genes encoding claudins and cadherins. This IOX2 effect was corroborated by an impaired barrier function, as measured by dextran permeability. These results provide a first insight into the effects associated with HIF1α stabilization in the respiratory epithelium, suggesting that HIF1α impacts properties that are key to maintaining homeostasis upon stimulation with a relevant bacterial agonist.

Indicadores sociodemográficos e de saneamento e moradia na qualidade de vida de pessoas com estomia / Sociodemographic and sanitation and housing indicators on the quality of life of people with stoma / Indicadores sociodemográficos y sanitarios y habitacionales sobre la calidad de vida de las personas ostomizadas

Objetivo:Analisar as relações entre a qualidade de vida (QV) de pessoas com estomia com indicadores sociodemográficos, clínicos, de estilo de vida, de saneamento e moradia. Métodos: Estudo transversal com amostra de 106 indivíduos com estomia entrevistados de maio a dezembro de 2019. Utilizou-se um questionário sociodemográfico e clínico e o City of Hope - Quality of Life - Ostomy Questionnary. Resultados: O bem-estar espiritual (7,71±1,09) foi o domínio com melhor performance. A QV não diferiu entre homens e mulheres (p = 0,372), porém esteve associada à escolaridade (< 0,001) e renda familiar (p = 0,025), ao diabetes (p = 0,008) e etilismo (p = 0,044), às condições da água para consumo (p < 0,001), ao destino do lixo (p = 0,021), em ter energia elétrica (p = 0,034), ao tipo de moradia (p = 0,026) e ao número de cômodos (p = 0,023) e tipo de cobertura da habitação (p = 0,021). Conclusão: Piores indicadores socioeconômicos, de saneamento básico e moradia, parecem impactar negativamente a QV de pessoas com estomias.

Objectives:To analyze the relationships between the quality of life (QOL) of people with stoma with sociodemographic, clinical, lifestyle, sanitation, and housing indicators. Method: Cross-sectional study with a sample of 106 individuals with stoma interviewed from May to December 2019. A sociodemographic and clinical questionnaire and the City of Hope Quality of Life Ostomy Questionnaire were used. Results: Spiritual well-being (7.71 ± 1.09) was the best performing domain. Quality of life did not differ between men and women (p = 0.372), but was associated with education (< 0.001) and family income (p = 0.025), diabetes (p = 0.008) and alcoholism (p = 0.044), drinking water conditions (p < 0.001), garbage disposal (p = 0.021), having electricity (p = 0.034), housing type (p = 0.026), number of rooms (p = 0.023), and housing coverage (p = 0.021). Conclusion: worse socioeconomic, sanitation, and housing indicators appear to negatively impact the QOL of people with stomata

Objetivo:Analizar las relaciones entre la calidad de vida (CV) de las personas con estoma con indicadores sociodemográficos, clínicos, de estilo de vida, saneamiento y vivienda. Métodos: Estudio transversal con una muestra de 106 ostomizados, entrevistados de mayo a noviembre de 2019. Se utilizó un cuestionario sociodemográfico y clínico y el Cuestionario City of Hope - Quality of Life - Ostomy. Resultados: El bienestar espiritual (7,71 ± 1,09) fue el dominio con mejor desempeño. La CV no difirió entre hombres y mujeres (p = 0,372), pero se asoció con la educación (< 0,001) y los ingresos familiares(p = 0,025), la diabetes (p = 0,008) y el alcoholismo (p = 0,044), con las condiciones de agua potable (p <0,001), el destino de la basura (p = 0,021), la disponibilidad de electricidad (p = 0,034), el tipo de vivienda (p = 0,026) y el número de habitaciones (p = 0,023) y tipo de cobertura de vivienda (p = 0,021). Conclusión: Peores indicadores socioeconómicos, saneamiento básico, vivienda, parecen impactar negativamente la calidad de vida de las personas con ostomía.

Combined Transcriptome and Proteome Leukocyte's Profiling Reveals Up-Regulated Module of Genes/Proteins Related to Low Density Neutrophils and Impaired Transcription and Translation Processes in Clinical Sepsis.

Sepsis is a global health emergency, which is caused by various sources of infection that lead to changes in gene expression, protein-coding, and metabolism. Advancements in "omics" technologies have provided valuable tools to unravel the mechanisms involved in the pathogenesis of this disease. In this study, we performed shotgun mass spectrometry in peripheral blood mononuclear cells (PBMC) from septic patients (N=24) and healthy controls (N=9) and combined these results with two public microarray leukocytes datasets. Through combination of transcriptome and proteome profiling, we identified 170 co-differentially expressed genes/proteins. Among these, 122 genes/proteins displayed the same expression trend. Ingenuity Pathway Analysis revealed pathways related to lymphocyte functions with decreased status, and defense processes that were predicted to be strongly increased. Protein-protein interaction network analyses revealed two densely connected regions, which mainly included down-regulated genes/proteins that were related to the transcription of RNA, translation of proteins, and mitochondrial translation. Additionally, we identified one module comprising of up-regulated genes/proteins, which were mainly related to low-density neutrophils (LDNs). LDNs were reported in sepsis and in COVID-19. Changes in gene expression level were validated using quantitative real-time PCR in PBMCs from patients with sepsis. To further support that the source of the upregulated module of genes/proteins found in our results were derived from LDNs, we identified an increase of this population by flow cytometry in PBMC samples obtained from the same cohort of septic patients included in the proteomic analysis. This study provides new insights into a reprioritization of biological functions in response to sepsis that involved a transcriptional and translational shutdown of genes/proteins, with exception of a set of genes/proteins related to LDNs and host-defense system.

Association between circulating exhausted CD4+ T cells with poor meningococcal C conjugate vaccine antibody response in HIV-infected children and adolescents.

OBJECTIVES: To investigate the expression levels of surface markers of activation (CD38 and HLA-DR), inhibition (PD-1, TIGIT and CD57) and co-stimulation (CD28 and CD127) on CD4+ T cells of children/adolescents with vertical HIV infection (HI patients) and HIV-uninfected (HU) controls vaccinated with the meningococcal C conjugate vaccine (MCC). METHODS: HI patients (n=12), aged 8-17 years, were immunized with two MCC injections, while HU controls (n=9), aged 5.3-10.7 years, received a single MCC dose (as per national recommendation at the time of this study, a single MCC vaccine dose should be given for healthy children and youth aged 1-18 years). The HI patients were categorized according to the combined antiretroviral therapy (cART) treatment. Blood samples were obtained before vaccination, after priming, and after the administration of a booster dose of vaccine to determine the serum bactericidal antibody (SBA) titers and the expression levels of surface markers on CD4+ T cells by flow cytometry. The levels of serum cytokines, IL-4 and CXCL-13 were also measured using Luminex kits. RESULTS: The co-expression of the TIGIT-HLA-DR-CD38 molecules increased in the CD4+ T cells of HI patients/no-cART who also showed a lower frequency of CD127+CD28+ CD4+ T cells than HI patients/cART and HU group subjects. There were significant negative correlations between the frequency of exhausted CD4+ T cells and the SBA response. IL-4 levels were higher in HI patients/cART and positively correlated with SBA titers but negatively associated with the expression of exhaustion markers. Moreover, the CXCL-13 levels were positively correlated with the exhausted CD4+ T cells. CONCLUSION: The results of our study suggest that the co-expression of exhaustion markers and/or loss of co-stimulatory molecules influence the SBA response in HI patients.

Post-treatment with the PPAR-γ agonist pioglitazone inhibits inflammation and bacterial growth during Klebsiella pneumonia.

Agonists of peroxisome proliferator-activated receptor (PPAR)-γ have been suggested as potential adjuvant therapy in bacterial pneumonia because of their capacity to inhibit inflammation and enhance bacterial clearance. Previous studies only assessed the effects of pretreatment with these compounds, thereby bearing less relevance for the clinical scenario. Moreover, PPAR-γ agonists have not been studied in pneumonia caused by Klebsiella pneumoniae, a common human respiratory pathogen of which antibiotic treatment is hampered by increasing antimicrobial resistance. Here we show that administration of the PPAR-γ agonist pioglitazone 6 or 8 h after infection of mice with a highly virulent strain of Klebsiella pneumoniae via the airways results in reduced cytokine and myeloperoxidase levels in the lungs at 24 h after infection, as well as reduced bacterial growth in the lungs and decreased dissemination to distant organs at 42 h post-infection. These results suggest that pioglitazone may be an interesting agent in the treatment of Klebsiella pneumonia.