RESUMO
La tuberculosis sigue siendo una epidemia mundial y en Chile no ha mostrado una tendencia descendente en los últimos años, con un aumento en los casos infantiles. En los niños, el diagnóstico es un reto debido a la baja carga bacilar y las características de las lesiones, que suelen ser cerradas. Métodos tradicionales como los cultivos, considerados anteriormente como el gold standard, con frecuencia arrojan resultados negativos. Sin embargo, los avances en pruebas moleculares han permitido un progreso significativo en la confirmación bacteriológica. Otras herramientas diagnósticas, como la prueba de tuberculina (PPD) y los ensayos de liberación de interferón gamma (IGRAs), tienen sensibilidades y especificidades variables, siendo útiles como pruebas complementarias. Las imágenes juegan un papel clave en la evaluación diagnóstica de tuberculosis pulmonar y extrapulmonar en pacientes pediátricos. Esta revisión aborda la epidemiología y el proceso diagnóstico de la tuberculosis infantil.
Tuberculosis remains a global epidemic, and in Chile, it has not shown a downward trend in recent years, with an increase in pediatric cases. Diagnosing tuberculosis in children presents challenges due to the low bacillary load and the closed nature of the lesions. Traditional methods like cultures, once considered the gold standard, often yield negative results. However, advances in molecular testing have significantly improved bacteriological confirmation. Other diagnostic tools, such as the tuberculin skin test (PPD) and interferon-gamma release assays (IGRAs), offer variable sensitivities and specificities and are useful as complementary tests. Imaging plays a critical role in the diagnostic evaluation of pulmonary and extrapulmonary tuberculosis in pediatric patients. This review addresses the epidemiology and diagnostic process of pediatric tuberculosis.
Assuntos
Humanos , Criança , Tuberculose/diagnóstico , Escarro/microbiologia , Tuberculose/genética , Tuberculose/microbiologia , Tuberculose/diagnóstico por imagem , Teste Tuberculínico , Técnicas de Diagnóstico Molecular , Testes de Liberação de Interferon-gama , Mycobacterium tuberculosis/isolamento & purificaçãoRESUMO
INTRODUÇÃO: A Cardiomiopatia Hipertrófica (CMH) é a cardiomiopatia hereditária mais prevalente no mundo, com amplo espectro de apresentação clínica. A despeito da diversidade de fenótipos da CMH, o acometimento de ventrículo direito (VD) é raro, com poucos casos descritos em literatura. Relatamos um caso raro de CMH de VD isolada em paciente atleta. RELATO DE CASO: EBO, masculino, 67 anos, atleta amador, assintomático, admitido para avaliação pré participação. Exame físico sem alterações. Eletrocardiograma (ECG) com bloqueio de ramo direito, sobrecarga ventricular esquerda e alteração de repolarização ventricular inferior. Realizado cintilografia com teste ergométrico, negativo para isquemia. Na fase de esforço, apresentou extrassístoles ventriculares monomórficas isoladas e frequentes. A ressonância cardíaca demonstrou aumento da espessura miocárdica na porção apical do VD (Figura 1-A) e realce tardio de padrão não coronariano no VD (Figura 1-B). Não foram registradas arritmias no Holter de 24h. Realizado painel molecular para hipertrofias ventriculares, que identificou variante patogênica no gene SLC22A5. Esse gene está relacionado a quadros de deficiência sistêmica primária de carnitina (DSPC). Caso foi discutido em reunião interdisciplinar e contraindicado esporte competitivo, com redução da intensidade do esforço físico realizado. CONCLUSÃO: Além da associação da variante patogênica no gene SLC22A5 com a DSPC, foram relatados casos de CMH de apresentação benigna e tardia. Entretanto, com a CMH de VD isolada nunca foi descrita na literatura. Concluímos ser fundamental a realização de painel molecular para confirmação do diagnóstico de CMH, exclusão de fenocópias e a valorização da variante patogênica no gene SCL22A5 no cenário da CMH.
Assuntos
Cardiomiopatias/genéticaRESUMO
Introdução: O trabalho conjunto da genética médica e da fonoaudiologia é essencial, contribuindo para o desenvolvimento de procedimentos que auxiliam no tratamento de pacientes com distúrbios da comunicação. Objetivo: Analisar as características fonoaudiológicas de pacientes pediátricos atendidos por um serviço de genética clínica. Método: Estudo transversal observacional, realizado com pacientes atendidos pelo serviço de genética de um hospital em Porto Alegre. Para a coleta de dados, aplicou-se um questionário relacionado as áreas de audição, deglutição, motricidade orofacial, voz e linguagem. Resultados: A amostra foi constituída por 54 participantes com idades entre 8 meses e 17 anos (média de idade 6 anos e 5 meses). 24,07% (n=13) dos pacientes apresentaram diagnóstico de síndrome, e 59,26% (n=32) tinham atraso no desenvolvimento neuropsicomotor. Com relação ao perfil fonoaudiológico, 81,48% (n=44) apresentaram algum hábito oral deletério durante a infância. 16,67% (n=9) percebiam alguma dificuldade para ouvir e 29,62% (n=16) para deglutir. 85,19% (n=46) dos participantes manifestaram a linguagem oral desenvolvida e, destes, 71,74% (n=33) apresentavam trocas na fala. 33,33% (n=18) já estavam em atendimento fonoaudiológico, e outros 24,07% (n=13) estavam na fila de espera para este atendimento. Conclusões: Uma parte significativa dos pacientes apresentou queixas e/ou manifestações nas áreas da comunicação humana, principalmente em relação à linguagem, à fala e aos hábitos orais deletérios. Esses dados destacam a importância do encaminhamento para a equipe de fonoaudiologia. (AU)
Introduction: The collaborative efforts of medical genetics and speech therapy are essential, contributing to the development of procedures that assist in treating patients with communication disorders. Objective: To analyze the speech therapy characteristics of pediatric patients seen by a clinical genetics service. Methods: Observational cross-sectional study conducted with patients seen at the genetics service of a hospital in Porto Alegre. A questionnaire related to hearing, swallowing, orofacial motricity, voice, and language areas was used for data collection. Results: The sample consisted of 54 participants aged between 8 months and 17 years, with an average age of 6 years and 5 months. 24.07% (n=13) of the patients had a diagnosis of syndrome, and 59.26% (n=32) had delayed neuropsychomotor development. Regarding the speech therapy profile, 81.48% (n=44) had some harmful oral habit during childhood. 16.67% (n=9) reported some difficulty in hearing, and 29.62% (n=16) in swallowing. 85.19% (n=46) of the participants showed developed oral language, and of these, 71.74% (n=33) made speech substitutions. 33.33% (n=18) of the patients were already undergoing speech therapy, and another 24.07% (n=13) were on the waiting list for this treatment. Conclusions: A significant portion of the patients presented complaints and/or manifestations in the areas of human communication, especially regarding language, speech, and harmful oral habits. These data highlight the importance of referral to the speech therapy team. (AU)
Introducción: La colaboración entre genética médica y foniatría es esencial para desarrollar procedimientos que ayuden en el tratamiento de pacientes con trastornos de la comunicación. Objetivo:Analizar las características de patología del habla y lenguaje de pacientes pediátricos atendidos por un servicio de genética clínica. Método: Estudio transversal observacional con pacientes atendidos por el servicio de genética de un hospital en Porto Alegre. Se aplicó un cuestionario sobre audición, deglución, motricidad orofacial, voz y lenguaje. Resultados: La muestra consistió en 54 participantes con edades comprendidas entre 8 meses y 17 años (media: 6 años y 5 meses). El 24,07% (n=13) de los pacientes tenían un diagnóstico de síndrome, y el 59,26% (n=32) presentaron retraso en el desarrollo neuropsicomotor. En cuanto al perfil foniatra, el 81,48% (n=44) presentaron algún hábito oral perjudicial durante la infancia. El 16,67% (n=9) reportaron dificultades para oír, y el 29,62% (n=16) para tragar. El 85,19% (n=46) manifestaron lenguaje oral desarrollado y, de ellos, el 71,74% (n=33) realizaban intercambios en el habla. El 33,33% (n=18) de los pacientes ya estaban en tratamiento foniatra y el 24,07% (n=13) estaban en lista de espera para este tratamiento. Conclusiones: Una parte significativa de los pacientes presentó quejas y/o manifestaciones en las áreas de la comunicación humana, especialmente en relación con el lenguaje, el habla y los hábitos orales perjudiciales, enfatizando la importancia de la derivación al equipo de foniatría. (AU)
Assuntos
Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Criança , Adolescente , Serviços de Saúde , Transtornos do Desenvolvimento da Linguagem/genética , Fonoterapia , Síndrome , Estudos Transversais , Inquéritos e Questionários , Genética Médica , Doenças Genéticas InatasRESUMO
The objective of this study was to evaluate the prevalence of human papillomavirus (HPV) genotypes and their relationship with different grades of cytological lesions in female students of the Faculty of Health Sciences of the National University of Chimborazo. Material and Methods: The research had a quantitative and descriptive approach, with a comparative analysis of HPV genotypes and cytological lesions in students of the Faculty of Health Sciences. It is an experimental and field study, cross-sectional and retrospective, conducted from November 2023 to March 2024. Thirty students were selected by quota sampling, analyzing conventional cytology and data using SPSS 26. The results showed that 75.8% of the samples had Bethesda Negative results, whereas 24.2% had some degree of cytological lesion (ASC-US 13.7%, L-SIL 8.1%, H-SIL 1.6%, and ASC-H 0.8%). Genotyping showed the high prevalence of HPV, with HPV 18 and 33 being the most common high-risk genotypes. The most common low-risk indicators were HPV 43 and 42. Conclusions: The study confirmed the high prevalence of HPV among female university students and established a significant correlation between high-risk genotypes and the presence of more severe cytological lesions. These findings underscore the need for interventions aimed at prevention and early treatment of HPV, especially in high-risk populations.
Assuntos
Genótipo , Papillomaviridae , Infecções por Papillomavirus , Estudantes , Humanos , Feminino , Infecções por Papillomavirus/virologia , Infecções por Papillomavirus/epidemiologia , Infecções por Papillomavirus/patologia , Estudantes/estatística & dados numéricos , Universidades , Estudos Transversais , Adulto Jovem , Papillomaviridae/genética , Papillomaviridae/isolamento & purificação , Adulto , Estudos Retrospectivos , Neoplasias do Colo do Útero/virologia , Neoplasias do Colo do Útero/epidemiologia , Neoplasias do Colo do Útero/patologia , Prevalência , Adolescente , Esfregaço Vaginal , Displasia do Colo do Útero/virologia , Displasia do Colo do Útero/epidemiologia , Displasia do Colo do Útero/patologiaRESUMO
Cannabis sativa can be classified in two main types, according to psychotropic cannabinoid ∆9-tetrahydrocannabinol (∆9-THC) content: the drug-type and the fiber-type. According to the European Monitoring Center for Drugs and Drug Addiction, most of the European Union countries consider the possession of cannabis, for personal use, a minor offense with possibility of incarceration. Despite of the model of legal supply (i.e., Spanish cannabis clubs, Netherlands coffee shops) or medical use (i.e., Italy), cannabis remains the most used and trafficked illicit plant in the European Union. Differentiating cannabis crops or tracing the biogeographical origin is crucial for law enforcement purposes. Chloroplast DNA (cpDNA) markers may assist to determine biogeographic origin and to differentiate hemp from marijuana. This research aims: to identify and to evaluate nine C. sativa cpDNA polymorphic SNP sites to differentiate crop type and to provide information about its biogeographical origin. Five SNaPshot™ assays for nine chloroplast markers were developed and conducted in marijuana samples seized in Chile, the USA-Mexico border and Spain, and hemp samples grown in Spain and in Italy. The SNapShot™ assays were tested on 122 cannabis samples, which included 16 blind samples, and were able to differentiate marijuana crop type from hemp crop type in all samples. Using phylogenetic analysis, genetic differences were observed between marijuana and hemp samples. Moreover, principal component analysis (PCA) supported the relationship among hemp samples, as well as for USA-Mexico border, Spanish, and Chilean marijuana samples. Genetic differences between groups based on the biogeographical origin and their crop type were observed. Increasing the number of genetic markers, including the most recently studied ones, and expanding the sample database will provide more accurate information about crop differentiation and biogeographical origin.
Assuntos
Cannabis , DNA de Cloroplastos , Polimorfismo de Nucleotídeo Único , Cannabis/genética , Marcadores Genéticos , DNA de Cloroplastos/genética , México , Reação em Cadeia da Polimerase , Europa (Continente) , Itália , Chile , EspanhaRESUMO
The novel HLA-DPB1*14:01:15 allele differs from DPB1*14:01:01:01 by change of C > T in exon 3.
Assuntos
Alelos , Sequência de Bases , Éxons , Cadeias beta de HLA-DP , Teste de Histocompatibilidade , Doadores de Tecidos , Humanos , Cadeias beta de HLA-DP/genética , Brasil , Análise de Sequência de DNA/métodos , Medula Óssea , Alinhamento de Sequência , Códon , Transplante de Medula ÓsseaRESUMO
Congenital Central Hypoventilation Syndrome (CCHS) is a rare genetic condition affecting the autonomic nervous system and respiratory center due to mutations in the PHOX2B gene, and it is associated with alveolar hypoventilation during sleep and sudden death. It requires early invasive mechanical ventilation (IMV). OBJECTIVE: To report a neonatal case successfully treated with non-invasive ventilatory support (NVS), avoiding tracheostomy. CLINICAL CASE: Full-term newborn, whose mother uses nocturnal NVS due to CCHS. During the transition period, she presented desaturations associated with hypercapnia and respiratory acidosis, without pulmonary involvement. She developed severe hypoventilation during sleep, with no respiratory effort, peripheral oxygen saturation (SpO2) < 80%, plus respiratory acidosis. While awake, she had good respiratory effort and normal SpO2 without assistance. Noninvasive continuous positive airway pressure and oxygen therapy worsened her condition while sleeping. Complete NVS with nasal interface and bi-level airway positive pressure, inspiratory/expiratory pressure 14-16/4 cm H2O, normalized SpO2 during sleep, and arterial blood gases while awake. Sequencing of the PHOX2B gene confirmed the presence of a heterozygous pathogenic variant with the 20/26 genotype. At 2 months of age, she was discharged maintaining NVS with nasal interface and 0 PEEP, achieving adequate neurodevelopment. CONCLUSION: We highlight the importance of genetic diagnosis of CCHS in neonates with clinical presentation of early alveolar hypoventilation, especially if there is a family history. We are not aware of other reports of neonatal onset in which NVS prevents IMV, in this potentially lethal pathology.
Assuntos
Proteínas de Homeodomínio , Hipoventilação , Apneia do Sono Tipo Central , Fatores de Transcrição , Humanos , Apneia do Sono Tipo Central/diagnóstico , Apneia do Sono Tipo Central/terapia , Apneia do Sono Tipo Central/genética , Recém-Nascido , Hipoventilação/congênito , Hipoventilação/terapia , Hipoventilação/diagnóstico , Hipoventilação/genética , Feminino , Proteínas de Homeodomínio/genética , Fatores de Transcrição/genética , Ventilação não Invasiva , Pressão Positiva Contínua nas Vias Aéreas , Acidose Respiratória/diagnóstico , Acidose Respiratória/terapia , Acidose Respiratória/etiologia , Mutação , OxigenoterapiaRESUMO
Mare endometrosis is a major reproductive problem associated with low fertility and is characterized by persistent inflammation, TGFß-1 signaling, and consequently, extracellular matrix deposition, which compromises endometrial glands. Mesenchymal stem cell-based products (MSCs), such as extracellular vesicles (EVs), have gained attention due to the regulatory effects exerted by their miRNA cargo. Here, we evaluated the impact of preconditioning equine adipose mesenchymal stem cells with TGFß-1 for short or long periods on the anti-fibrotic properties of secreted extracellular vesicles. MSCs were isolated from six healthy horses and exposed to TGFß-1 for 4, 24, and 0 h. The expression of anti-fibrotic and pro-fibrotic miRNAs and mRNAs in treated cells and miRNAs in the cargo of secreted extracellular vesicles was measured. The resulting EVs were added for 48 h to endometrial stromal cells previously induced to a fibrotic status. The expression of anti-fibrotic and pro-fibrotic genes and miRNAs was evaluated in said cells using qPCR and next-generation sequencing. Preconditioning MSCs with TGFß-1 for 4 h enriched the anti-fibrotic miRNAs (mir29c, mir145, and mir200) in cells and EVs. Conversely, preconditioning the cells for 24 h leads to a pro-fibrotic phenotype overexpressing mir192 and mir433. This finding might have implications for developing an EV-based protocol to treat endometrial fibrosis in mares.
Assuntos
Endométrio , Vesículas Extracelulares , Fibrose , Células-Tronco Mesenquimais , MicroRNAs , Fator de Crescimento Transformador beta1 , Animais , Cavalos , Feminino , Células-Tronco Mesenquimais/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Vesículas Extracelulares/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta1/genética , Endométrio/metabolismo , Endométrio/citologia , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Células Estromais/metabolismo , Células Estromais/efeitos dos fármacos , Doenças dos Cavalos , Regulação da Expressão Gênica/efeitos dos fármacos , Endometriose/veterinária , Endometriose/metabolismo , Endometriose/genéticaRESUMO
Hydrogen peroxide (H2O2) transport by aquaporins (AQP) is a critical feature for cellular redox signaling. However, the H2O2 permeation mechanism through these channels remains poorly understood. Through functional assays, two Plasma membrane Intrinsic Protein (PIP) AQP from Medicago truncatula, MtPIP2;2 and MtPIP2;3 have been identified as pH-gated channels capable of facilitating the permeation of both water (H2O) and H2O2. Employing a combination of unbiased and enhanced sampling molecular dynamics simulations, we investigated the key barriers and translocation mechanisms governing H2O2 permeation through these AQP in both open and closed conformational states. Our findings reveal that both H2O and H2O2 encounter their primary permeation barrier within the selectivity filter (SF) region of MtPIP2;3. In addition to the SF barrier, a second energetic barrier at the NPA (asparagine-proline-alanine) region that is more restrictive for the passage of H2O2 than for H2O, was found. This behavior can be attributed to a dissimilar geometric arrangement and hydrogen bonding profile between both molecules in this area. Collectively, these findings suggest mechanistic heterogeneity in H2O and H2O2 permeation through PIPs.
Assuntos
Aquaporinas , Peróxido de Hidrogênio , Simulação de Dinâmica Molecular , Proteínas de Plantas , Água , Peróxido de Hidrogênio/metabolismo , Aquaporinas/metabolismo , Aquaporinas/química , Aquaporinas/genética , Água/metabolismo , Água/química , Proteínas de Plantas/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/genética , Medicago truncatula/metabolismo , Medicago truncatula/genética , Membrana Celular/metabolismo , Ligação de HidrogênioAssuntos
Melanoma , Osteopontina , Humanos , Melanoma/genética , Melanoma/diagnóstico , Osteopontina/genética , Osteopontina/metabolismo , Prognóstico , Regulação Neoplásica da Expressão Gênica , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Neoplasias Cutâneas/genética , Neoplasias Cutâneas/diagnóstico , Neoplasias Cutâneas/patologia , Processamento Alternativo , Feminino , MasculinoRESUMO
Monoxenous trypanosomatid Strigomonas culicis harbors an endosymbiotic bacterium, which enables the protozoa to survive without heme supplementation. The impact of H2O2 resistance and symbiont elimination on intracellular heme and Fe2+ availability was analyzed through a comparison of WT strain with both WT H2O2-resistant (WTR) and aposymbiotic (Apo) protozoa. The relative quantification of the heme biosynthetic pathway through label-free parallel reaction monitoring targeted mass spectrometry revealed that H2O2 resistance does not influence the abundance of tryptic peptides. However, the Apo strain showed increased coproporphyrinogen III oxidase and ferrochelatase levels. A putative ferrous iron transporter, homologous to LIT1 and TcIT from Leishmania major and Trypanosoma cruzi, was identified for the first time. Label-free parallel reaction monitoring targeted mass spectrometry also showed that S. culicis Iron Transporter (ScIT) increased 1.6- and 16.4-fold in WTR and Apo strains compared to WT. Accordingly, antibody-mediated blockage of ScIT decreased by 28.0% and 40.0% intracellular Fe2+concentration in both WTR and Apo strains, whereas no effect was detected in WT. In a heme-depleted medium, adding 10 µM hemin decreased ScIT transcript levels in Apo, whereas 10 µM PPIX, the substrate of ferrochelatase, increased intracellular Fe2+ concentration and ferric iron reduction. Overall, the data suggest mechanisms dependent on de novo heme synthesis (and its substrates) in the Apo strain to overcome reduced heme availability. Given the importance of heme and Fe2+ as cofactors in metabolic pathways, including oxidative phosphorylation and antioxidant systems, this study provides novel mechanistic insights associated with H2O2 resistance in S. culicis.
Assuntos
Heme , Peróxido de Hidrogênio , Simbiose , Heme/metabolismo , Peróxido de Hidrogênio/metabolismo , Trypanosomatina/metabolismo , Trypanosomatina/genética , Ferro/metabolismo , Resistência a Medicamentos , Proteínas de Protozoários/metabolismo , Proteínas de Protozoários/genéticaRESUMO
The aim of this study was to evaluate whether polymorphisms in SOD2 and SOD3 genes modulate the oral health-related quality of life (OHRQoL) of Para athletes with dental caries experience. The cross-sectional study included 264 Para athletes (143 in athletics, 61 in weightlifting and 60 in swimming). A trained and calibrated team recorded the decayed, missing and filled teeth index (DMFT). The Brazilian version of the Oral Health Impact Profile (OHIP-14) was used to measure OHRQoL. Genomic DNA was extracted from the athletes' saliva, and genetic polymorphisms in the SOD2 (rs5746136 and rs10370) and SOD3 (rs2855262 and rs13306703) genes were analyzed by real-time polymerase chain reaction. Univariate and multivariate analyses were performed. A multivariate General Linear Model analysis, adjusted for sex, revealed that the SOD3 gene polymorphism (rs2855262) had a significant effect on the psychological disability domain [codominant (p = 0.045) and recessive (p=0.038) models]. The SOD2 gene polymorphism (rs5746136) had a significant effect on the total OHIP-14 score [dominant model (p = 0.038)] and the psychological discomfort [dominant model (p = 0.034)] and physical disability [codominant model (p=0.037)] domains. Presence of the SOD2 rs10370 polymorphism led to statistical differences in the total score [codominant (p = 0.026) and dominant (p = 0.023) models] and the handicap domain scores [codominant (p = 0.027) and dominant (p = 0.032) models]. Polymorphisms of the SOD2 and SOD3 genes may be important biomarkers of OHRQoL in Para athletes with dental caries experience.
Assuntos
Atletas , Saúde Bucal , Qualidade de Vida , Superóxido Dismutase , Adolescente , Adulto , Feminino , Humanos , Masculino , Adulto Jovem , Análise de Variância , Atletas/psicologia , Atletas/estatística & dados numéricos , Brasil , Estudos Transversais , Cárie Dentária/genética , Índice CPO , Polimorfismo Genético/genética , Polimorfismo de Nucleotídeo Único , Reação em Cadeia da Polimerase em Tempo Real , Valores de Referência , Saliva/química , Estatísticas não Paramétricas , Superóxido Dismutase/genéticaRESUMO
A T-cell-independent (TI) pathway activated by microbiota results in the generation of low-affinity homeostatic IgA with a critical role in intestinal homeostasis. Moderate aerobic exercise (MAE) provides a beneficial impact on intestinal immunity, but the action of MAE on TI-IgA generation under senescence conditions is unknown. This study aimed to determine the effects of long-term MAE on TI-IgA production in young (3 month old) BALB/c mice exercised until adulthood (6 months) or aging (24 months). Lamina propria (LP) from the small intestine was obtained to determine B cell and plasma cell sub-populations by flow cytometry and molecular factors related to class switch recombination [Thymic Stromal Lymphopoietin (TSLP), A Proliferation-Inducing Ligand (APRIL), B Cell Activating Factor (BAFF), inducible nitric oxide synthase (iNOS), and retinal dehydrogenase (RDH)] and the synthesis of IgA [α-chain, interleukin (IL)-6, IL-21, and Growth Factor-ß (TGF-ß)]; and epithelial cells evaluated IgA transitosis [polymeric immunoglobulin receptor (pIgR), tumor necrosis factor-α (TNF-α), interferon-γ (IFN-γ), IL-4] by the RT-qPCR technique. The results were compared with data obtained from sedentary age-matched mice. Statistical analysis was computed with ANOVA, and p < 0.05 was considered to be a statistically significant difference. Under senescence conditions, MAE promoted the B cell and IgA+ B cells and APRIL, which may improve the intestinal response and ameliorate the inflammatory environment associated presumably with the downmodulation of pro-inflammatory mediators involved in the upmodulation of pIgR expression. Data suggested that MAE improved IgA and downmodulate the cytokine pro-inflammatory expression favoring homeostatic conditions in aging.
Assuntos
Envelhecimento , Homeostase , Imunoglobulina A , Camundongos Endogâmicos BALB C , Condicionamento Físico Animal , Animais , Imunoglobulina A/metabolismo , Imunoglobulina A/imunologia , Camundongos , Envelhecimento/imunologia , Citocinas/metabolismo , Linfócitos B/imunologia , Linfócitos B/metabolismo , Fator Ativador de Células B/metabolismo , Fator Ativador de Células B/genética , Mucosa Intestinal/metabolismo , Mucosa Intestinal/imunologia , Intestino Delgado/imunologia , Intestino Delgado/metabolismo , Masculino , Plasmócitos/imunologia , Plasmócitos/metabolismo , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/metabolismo , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral/genéticaRESUMO
Cildáñez stream (in Matanza-Riachuelo basin, Buenos Aires) is one of the most polluted watercourses of Argentina, containing a mixed contamination from agricultural and industrial wastes. The application of water bioremediation processes for this kind of effluent will require microorganisms with a high tolerance to contamination. In this sense, obtaining higher contaminant-resistant microalgae lines is widely desired. In this study, adaptive laboratory evolution (ALE) and random mutagenesis were used to obtain Chlorella vulgaris LMPA-40 strains adapted to grow in polluted water from the Cildáñez stream. The ALE process was performed by 22 successive subcultures under selective pressure (Cildáñez wastewater alone or with the addition of phenol or H2O2) while random mutagenesis was performed with UV-C radiation at 275nm. Not all the cell lines obtained after ALE could adapt enough to overcome the stress caused by the Cildáñez wastewater, indicating that the process is quite random and depends on the stressor used. The best results were obtained for the Cildáñez wastewater adapted cells (Cild 3 strain) that were more resistant than the original strain. The concentration of protein, Chlorophyll A, Chlorophyll B, and carotenoids in the Cild 3 ALE evolved strain was higher than that of the control strain. However, this strain exhibited half of the lipid content compared to the same control strain. Interestingly, these alterations and the acquired tolerance may be reversed over time during storage. These findings suggest that the acquisition of novel cell lines could not be permanent, a fact that must be considered for future trials.
Assuntos
Chlorella vulgaris , Chlorella vulgaris/genética , Águas Residuárias/microbiologia , Argentina , Biodegradação Ambiental , Evolução Molecular Direcionada , Mutagênese , Clorofila A , Clorofila/análise , Peróxido de Hidrogênio/farmacologiaRESUMO
Sexual dimorphism among mammals includes variations in the pain threshold. These differences are influenced by hormonal fluctuations in females during the estrous and menstrual cycles of rodents and humans, respectively. These physiological conditions display various phases, including proestrus and diestrus in rodents and follicular and luteal phases in humans, distinctly characterized by varying estrogen levels. In this study, we evaluated the capsaicin responses in male and female mice at different estrous cycle phases, using two murine acute pain models. Our findings indicate that the capsaicin-induced pain threshold was lower in the proestrus phase than in the other three phases in both pain assays. We also found that male mice exhibited a higher pain threshold than females in the proestrus phase, although it was similar to females in the other cycle phases. We also assessed the mRNA and protein levels of TRPV1 in the dorsal root and trigeminal ganglia of mice. Our results showed higher TRPV1 protein levels during proestrus compared to diestrus and male mice. Unexpectedly, we observed that the diestrus phase was associated with higher TRPV1 mRNA levels than those in both proestrus and male mice. These results underscore the hormonal influence on TRPV1 expression regulation and highlight the role of sex steroids in capsaicin-induced pain.
Assuntos
Capsaicina , Dor , Canais de Cátion TRPV , Animais , Canais de Cátion TRPV/metabolismo , Canais de Cátion TRPV/genética , Capsaicina/farmacologia , Masculino , Feminino , Camundongos , Dor/metabolismo , Dor/genética , Hormônios Esteroides Gonadais/metabolismo , Ciclo Estral/efeitos dos fármacos , Limiar da Dor/efeitos dos fármacos , Gânglios Espinais/metabolismo , Gânglios Espinais/efeitos dos fármacos , Gânglio Trigeminal/metabolismo , Gânglio Trigeminal/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Caracteres Sexuais , RNA Mensageiro/metabolismo , RNA Mensageiro/genéticaRESUMO
Advanced glycated end products (AGEs) are cytotoxic compounds that are mainly increased in diabetes mellitus (DM), kidney failure, inflammation, and in response to the ingestion of AGE-rich diets. AGEs can also impair glycemic homeostasis by decreasing the expression of the Slc2a4 (solute carrier family 2 member 4) gene and its GLUT4 (solute carrier family 2, facilitated glucose transporter member 4) protein in muscle. However, the mechanisms underlying AGE's effect on adipocytes have not been demonstrated yet. This study investigated the effects of AGEs upon Slc2a4/GLUT4 expression in 3T3-L1 adipocytes, as well as the potential role of NFKB (nuclear factor NF-kappa-B) activity in the effects observed. Adipocytes were cultured in the presence of control albumin (CA) or advanced glycated albumin (GA) at concentrations of 0.4, 3.6, and 5.4 mg/mL for 24 h or 72 h. Slc2a4, Rela, and Nfkb1mRNAs were measured by RT-qPCR, GLUT4, IKKA/B, and p50/p65 NFKB subunits using Western blotting, and p50/p65 binding into the Slc2a4 promoter was analyzed by chromatin immunoprecipitation (ChIP) assay. GA at 0.4 mg/mL increased Slc2a4/GLUT4 expression after 24 h and 72 h (from 50% to 100%), but at 5.4 mg/mL, Slc2a4/GLUT4 expression decreased at 72 h (by 50%). Rela and Nfkb1 expression increased after 24 h at all concentrations, but this effect was not observed at 72 h. Furthermore, 5.4 mg/mL of GA increased the p50/p65 nuclear content and binding into Slc2a4 at 72 h. In summary, this study reveals AGE-induced and NFKB-mediated repression of Slc2a4/GLUT4 expression. This can compromise the adipocyte glucose utilization, contributing not only to the worsening of glycemic control in DM subjects but also the impairment of glycemic homeostasis in non-DM subjects under the high intake of AGE-rich foods.
Assuntos
Células 3T3-L1 , Adipócitos , Transportador de Glucose Tipo 4 , Produtos Finais de Glicação Avançada , Fator de Transcrição RelA , Animais , Camundongos , Adipócitos/metabolismo , Adipócitos/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Transportador de Glucose Tipo 4/metabolismo , Transportador de Glucose Tipo 4/genética , Produtos Finais de Glicação Avançada/metabolismo , Produtos Finais de Glicação Avançada/farmacologia , NF-kappa B/metabolismo , Subunidade p50 de NF-kappa B/metabolismo , Subunidade p50 de NF-kappa B/genética , Regiões Promotoras Genéticas , Fator de Transcrição RelA/metabolismo , Fator de Transcrição RelA/genéticaRESUMO
It is well established that microRNA-21 (miR-21) targets phosphatase and tensin homolog (PTEN), facilitating epithelial-to-mesenchymal transition (EMT) and drug resistance in cancer. Recent evidence indicates that PTEN activates its pseudogene-derived long non-coding RNA, PTENP1, which in turn inhibits miR-21. However, the dynamics of PTEN, miR-21, and PTENP1 in the DNA damage response (DDR) remain unclear. Thus, we propose a dynamic Boolean network model by integrating the published literature from various cancers. Our model shows good agreement with the experimental findings from breast cancer, hepatocellular carcinoma (HCC), and oral squamous cell carcinoma (OSCC), elucidating how DDR activation transitions from the intra-S phase to the G2 checkpoint, leading to a cascade of cellular responses such as cell cycle arrest, senescence, autophagy, apoptosis, drug resistance, and EMT. Model validation underscores the roles of PTENP1, miR-21, and PTEN in modulating EMT and drug resistance. Furthermore, our analysis reveals nine novel feedback loops, eight positive and one negative, mediated by PTEN and implicated in DDR cell fate determination, including pathways related to drug resistance and EMT. Our work presents a comprehensive framework for investigating cellular responses following DDR, underscoring the therapeutic potential of targeting PTEN, miR-21, and PTENP1 in cancer treatment.
Assuntos
Dano ao DNA , Resistencia a Medicamentos Antineoplásicos , Transição Epitelial-Mesenquimal , MicroRNAs , PTEN Fosfo-Hidrolase , RNA Longo não Codificante , PTEN Fosfo-Hidrolase/metabolismo , PTEN Fosfo-Hidrolase/genética , MicroRNAs/genética , MicroRNAs/metabolismo , Humanos , Transição Epitelial-Mesenquimal/genética , Resistencia a Medicamentos Antineoplásicos/genética , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patologia , Neoplasias/tratamento farmacológico , Linhagem Celular Tumoral , Apoptose/efeitos dos fármacos , Apoptose/genética , Transdução de SinaisRESUMO
OBJECTIVE: to report the first case series of Brazilian children diagnosed with Kleefstra syndrome, present a possible phenotype expansion to the syndrome and to raise physicians' awareness for this rare disease. RESULTS: seven patients with confirmed KS were evaluated, including 5 males and 2 females. Abnormal prenatal findings were observed in 4 patients. Most patients were born at term, with normal birth measurements. All patients had neurodevelopmental delay and 6 evolved with intellectual disability. Hearing loss was present in 57.1% of patients and 28.7% had congenital heart disease. In males, cryptorchidism was present in 75%. Despite the facial dysmorphisms, only 2 out of 7 patients had a pre-test clinical suspicion of KS. One specific patient presented bilateral agenesis of the semicircular canals, a very rare ear manifestation in Kleefstra syndrome, representing a possible phenotype expansion of the syndrome. CONCLUSION: this report aims to promote awareness among physicians evaluating patients in a context of neurodevelopmental delay or congenital malformations, especially congenital heart defects. We also highlight a possible phenotype expansion of the syndrome, with a case of semicircular anomaly, not reported in this syndrome so far.
Assuntos
Deleção Cromossômica , Anormalidades Craniofaciais , Deficiência Intelectual , Fenótipo , Canais Semicirculares , Humanos , Masculino , Feminino , Deficiência Intelectual/genética , Deficiência Intelectual/patologia , Brasil , Anormalidades Craniofaciais/patologia , Anormalidades Craniofaciais/genética , Criança , Pré-Escolar , Canais Semicirculares/anormalidades , Canais Semicirculares/patologia , Cardiopatias Congênitas/patologia , Cardiopatias Congênitas/genética , Cromossomos Humanos Par 9/genética , LactenteRESUMO
BACKGROUND: Exercise-induced pulmonary haemorrhage (EIPH) in athletic horses is characterized by the presence of blood from the lungs in the tracheobronchial tree after intense exercise. Despite the high prevalence of EIPH in horses, the primary aetiology remains unknown. Variants in the genes encoding CD39 and CD39L1 (ENTPD1 and ENTPD2, respectively) were previously reported as potential genetic causes involved in EIPH pathogenesis. However, the role of these variants in haemostatic functions is unknown. RESULTS: To investigate the association between EIPH and missense variants in the ENTPD1 (rs1152296272, rs68621348, and rs68621347) and ENTPD2 genes (rs782872967), 76 Thoroughbred horses diagnosed with EIPH and 56 without clinical signs of EIPH (control group) by trachea-bronchial endoscopy were genotyped. The rs1152296272 and rs68621347 variants were linked, which explained why the same results were found in all horses. Approximately 96% and 95% of the EIPH and control horses, respectively, carried at least one nonreference allele for these variants. In contrast, 100% of the control horses and 96% of the EIPH horses were homozygous for the reference allele for the rs68621348 variant. In the EIPH group, 1.5% of the horses were homozygotes and 24% were heterozygous for the nonreference allele of the rs782872967 variant. In the control group, the nonreference allele of this variant was observed only in heterozygotes (16%). There were no significant differences between groups for any of the variants. CONCLUSIONS: The variants previously described in the genes encoding the CD39 and CD39L1 enzymes were highly present in the studied population. However, no association was found between the occurrence of EIPH and the presence of these variants in Thoroughbred horses in this study.
Assuntos
Hemorragia , Doenças dos Cavalos , Pneumopatias , Condicionamento Físico Animal , Animais , Cavalos , Doenças dos Cavalos/genética , Hemorragia/veterinária , Hemorragia/genética , Pneumopatias/veterinária , Pneumopatias/genética , Masculino , Apirase/genética , Feminino , Predisposição Genética para Doença , Genótipo , Mutação de Sentido IncorretoRESUMO
INTRODUCTION: Producing commercial bacterins/toxoids against Clostridium spp. is laborious and hazardous. Conversely, developing prototype vaccines using purified recombinant toxoids, though safe and effective, is both laborious and costly for application in production animals. OBJECTIVE: Considering that inactivated recombinant Escherichiacoli (bacterin) is a simple, cost-effective, and to be safe solution, we evaluated, for the first time, a pentavalent formulation of recombinant bacterins containing the alpha, beta, and epsilon toxins of Clostridiumperfringens and C and D neurotoxins of Clostridiumbotulinum in sheep. METHODS: Subcutaneously, 18 Texel sheep received two doses (200 µg of each antigen) of recombinant bacterin (n = 7) or purified recombinant antigens (n = 6) on days 0 and 28, while the control group (n = 5) did not receive an immunization. Sera samples from days 0 (before the 1st dose), 28 (before the 2nd dose), and 56, 84, and 112 were used for measuring IgG (indirect ELISA) and neutralizing antibodies (mouse serum neutralization). RESULTS: Both formulations induced significant levels of IgG against all five toxins (p < 0.05) up to day 112, with peaks at days 28 and 56 post-immunization. The expected booster effect occurred only for the botulinum toxins. The neutralizing antibody titers were satisfactory against ETX (≥2 IU/ml for both formulations) and BoNT-D [5 IU/ml (bacterin) and 10 IU/ml (purified)]. CONCLUSION: While adjustments are required, the recombinant bacterin platform holds great potential for polyvalent vaccines due to its straightforward, safe, and cost-effective production, establishing it as a user-friendly technology for the veterinary immunobiological industry.