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Aquaculture fish are kept for long periods in sea cages or tanks. Consequently, accumulated stress causes the fish to present serious problems with critical economic losses. Fish food has been supplemented to reduce this stress, using many components as amino acids such as tryptophan. This study aims to determine the transcriptional effect of tryptophan and cortisol on primary cell cultures of salmon head and posterior kidney. Our results indicate activation of the kynurenine pathway and serotonin activity when stimulated with tryptophan and cortisol. An amount of 95% of tryptophan is degraded by the kynurenine pathway, indicating the relevance of knowing how this pathway is activated and if stress levels associated with fish culture trigger its activation. Additionally, it is essential to know the consequence of increasing kynurenic acid "KYNA" levels in the short and long term, and even during the fish ontogeny.
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Nutritional immunity regulates the homeostasis of micronutrients such as iron, manganese, and zinc at the systemic and cellular levels, preventing the invading microorganisms from gaining access and thereby limiting their growth. Therefore, the objective of this study was to evaluate the activation of nutritional immunity in specimens of Atlantic salmon (Salmo salar) that are intraperitoneally stimulated with both live and inactivated Piscirickettsia salmonis. The study used liver tissue and blood/plasma samples on days 3, 7, and 14 post-injections (dpi) for the analysis. Genetic material (DNA) of P. salmonis was detected in the liver tissue of fish stimulated with both live and inactivated P. salmonis at 14 dpi. Additionally, the hematocrit percentage decreased at 3 and 7 dpi in fish stimulated with live P. salmonis, unchanged in fish challenged with inactivated P. salmonis. On the other hand, plasma iron content decreased during the experimental course in fish stimulated with both live and inactivated P. salmonis, although this decrease was statistically significant only at 3 dpi. Regarding the immune-nutritional markers such as tfr1, dmt1, and ireg1 were modulated in the two experimental conditions, compared to zip8, ft-h, and hamp, which were down-regulated in fish stimulated with live and inactivated P. salmonis during the course experimental. Finally, the intracellular iron content in the liver increased at 7 and 14 dpi in fish stimulated with live and inactivated P. salmonis, while the zinc content decreased at 14 dpi under both experimental conditions. However, stimulation with live and inactivated P. salmonis did not alter the manganese content in the fish. The results suggest that nutritional immunity does not distinguish between live and inactivated P. salmonis and elicits a similar immune response. Probably, this immune mechanism would be self-activated with the detection of PAMPs, instead of a sequestration and/or competition of micronutrients by the living microorganism.
Assuntos
Piscirickettsia , Salmo salar , Animais , Manganês , Piscirickettsia/genética , FerroRESUMO
In this article, we analyze the internal structure of the scale for experience in exercising the right to health care (EERHC), based on the focus from the World Health Organization (WHO) on the right to health care, from the perspective of international migrants, in Chile. The methodology was an instrumental study (n = 563) conducted to analyze the psychometric properties of the EERHC scale. Its reliability and internal consistency were evaluated, while the exploratory structural equation modeling (ESEM) model and confirmatory factor analysis (CFA) were used to identify the structure of relationships between the variables measured. The item-dimension correlations obtained present levels of r ≥ 0.3, and the Cronbach's α and McDonald's ω presented ranges >0.9, considered to be acceptable on all models. Results: the model was selected for presenting a good fit index χ2 = 24,850, df = 300, p = 0.000; RMSEA = 0.07; CFI = 0.97; TLI = 0.95; and SRMR = 0.03. The evidence obtained lets us conclude that the scale has forty-five items and four dimensions. The findings demonstrate a good internal structure and are useful to measure primary health care service utilization based on the framework.
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OBJECTIVES: Xerostomia in SS patients has been associated with low quality and quantity of salivary mucins, which are fundamental for the hydration and protection of the oral mucosa. The aim of this study was to evaluate if cytokines induce aberrant mucin expression and whether tauroursodeoxycholic acid (TUDCA) is able to counteract such an anomaly. METHODS: Labial salivary glands from 16 SS patients and 15 control subjects, as well as 3D acini or human submandibular gland cells stimulated with TNF-α or IFN-γ and co-incubated with TUDCA, were analysed. mRNA and protein levels of Mucin 1 (MUC1) and MUC7 were determined by RT-qPCR and western blot, respectively. Co-immunoprecipitation and immunofluorescence assays for mucins and GRP78 [an endoplasmic reticulum (ER)-resident protein] were also performed. mRNA levels of RelA/p65 (nuclear factor-κB subunit), TNF-α, IL-1ß, IL-6, SEL1L and EDEM1 were determined by RT-qPCR, and RelA/p65 localization was evaluated by immunofluorescence. RESULTS: MUC1 is overexpressed and accumulated in the ER of labial salivary gland from SS patients, while MUC7 accumulates throughout the cytoplasm of acinar cells; however, MUC1, but not MUC7, co-precipitated with GRP78. TUDCA diminished the overexpression and aberrant accumulation of MUC1 induced by TNF-α and IFN-γ, as well as the nuclear translocation of RelA/p65, together with the expression of inflammatory and ER stress markers in 3D acini. CONCLUSION: Chronic inflammation alters the secretory process of MUC1, inducing ER stress and affecting the quality of saliva in SS patients. TUDCA showed anti-inflammatory properties decreasing aberrant MUC1 accumulation. Further studies are necessary to evaluate the potential therapeutic effect of TUDCA in restoring glandular homeostasis in SS patients.
Assuntos
Células Acinares/efeitos dos fármacos , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Mucina-1/efeitos dos fármacos , Glândulas Salivares Menores/efeitos dos fármacos , Síndrome de Sjogren/metabolismo , Glândula Submandibular/efeitos dos fármacos , Ácido Tauroquenodesoxicólico/farmacologia , Xerostomia/metabolismo , Células Acinares/metabolismo , Adulto , Idoso , Estudos de Casos e Controles , Células Cultivadas , Chaperona BiP do Retículo Endoplasmático , Estresse do Retículo Endoplasmático/genética , Feminino , Proteínas de Choque Térmico/efeitos dos fármacos , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Humanos , Imunoprecipitação , Técnicas In Vitro , Interferon gama/farmacologia , Interleucina-1beta/efeitos dos fármacos , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Proteínas de Membrana/efeitos dos fármacos , Proteínas de Membrana/genética , Proteínas de Membrana/metabolismo , Pessoa de Meia-Idade , Mucina-1/genética , Mucina-1/metabolismo , Mucinas/efeitos dos fármacos , Mucinas/genética , Mucinas/metabolismo , Proteínas/efeitos dos fármacos , Proteínas/genética , Proteínas/metabolismo , RNA Mensageiro/efeitos dos fármacos , RNA Mensageiro/metabolismo , Glândulas Salivares Menores/metabolismo , Proteínas e Peptídeos Salivares/efeitos dos fármacos , Proteínas e Peptídeos Salivares/genética , Proteínas e Peptídeos Salivares/metabolismo , Síndrome de Sjogren/genética , Glândula Submandibular/citologia , Glândula Submandibular/metabolismo , Fator de Transcrição RelA/efeitos dos fármacos , Fator de Transcrição RelA/genética , Fator de Transcrição RelA/metabolismo , Fator de Necrose Tumoral alfa/efeitos dos fármacos , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/farmacologia , Xerostomia/genéticaRESUMO
Pathogen interactions with cultured fish populations are well studied, but their effects on native fishes have not been characterized. In Chile, the disease caused by bacterial species Piscirickettsia salmonis represents one of the main issues and is considered to be one of the important pathogens in the field of aquaculture. They have been found to infect native fish. Therefore, it is necessary to understand the impact of P. salmonis on native species of local commercial value, as well as the potential impact associated with the emergence of antibiotic-resistant strains of P. salmonis. Due to this purpose, the native fish Eleginops maclovinus was used in our study. Fish were randomly distributed in tanks and intraperitoneally inoculated with two strains of P. salmonis. No mortality was recorded during the experiment. Cortisol, glucose and total α-amino acid levels increased in fish injected with AUSTRAL-005 strain compared to sham-injected and LF-89-inoculated fish. Moreover, results showed an increase in the activity of carbohydrates and lipids metabolism in liver; and an increase in the carbohydrates, lipids and total α-amino acid metabolism in muscle after injection with AUSTRAL-005. Our results suggest that P. salmonis modulates the physiology of E. maclovinus and the physiological impact increase in the presence of the antibiotic-resistant strain AUSTRAL-005.
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Doenças dos Peixes/microbiologia , Perciformes , Piscirickettsia/fisiologia , Infecções por Piscirickettsiaceae/veterinária , Transcrição Gênica , Animais , Regiões Antárticas , Chile , Infecções por Piscirickettsiaceae/microbiologiaRESUMO
INTRODUCCIÓN: La Infección Necrotizante de tejidos blandos (INTB) tiene una elevada morbimortalidad. El objetivo de este trabajo es describir del manejo perioperatorio de menores de 15 años que cursaron con INTB durante 15 años en un Hospital pediátrico. MATERIAL Y MÉTODOS: serie de pacientes identificados INTB entre 2000 y 2015 en el Hospital Roberto del Río. Se describen variables demográficas, clínicas, vacuna, tratamiento, cirugías, complicaciones, microorganismos, seguimiento, y fallecimientos. RESULTADOS: 22 pacientes, con mediana de 2 años y 9 meses de edad. 50% estaban cursando con una varicela. Dos fallecieron. Ninguno era previamente vacunado contra el virus varicela zoster. La localización fue tronco (14), extremidades (7), cuello (1). Los pacientes conscientes presentaron hiperestesia cutánea. Se realizó aseo quirúrgico con una mediana de 6,8 horas desde el inicio de la hiperestesia y 2 horas desde la sospecha diagnóstica. Los microorganismos fueron: S. pyogenes (38%) y E. coli (31%). Los antibióticos más frecuentes fueron penicilina más clindamicina. La herida se manejó con cierre primario, injertos y/o colgajos. Cinco evolucionaron con secuelas que necesitaron tratamiento. CONCLUSIÓN: Se identificó que la INTB puede presentarse con varicela. Debe sospecharse en lesiones cutáneas e hiperestesia desproporcionada. Resección amplia y precoz son fundamentales para el tratamiento.
INTRODUCTION: Necrotizing soft tissue infection (NSTI) has a high morbidity and mortality. The objective of this study is to describe the perioperative management of children under 15 years of age who have had this condition for 15 years in a pediatric hospital. MATERIAL AND METHODS: cases series of patients identified by Pathological Anatomy with NSTI during 2000 and 2015 at the Dr. Roberto del Río Hospital. Demographic variables, treatment, antecedents of vaccination, surgeries, complications, microorganisms, follow-up, and deaths are described. RESULTS: 22 patients were identified, with a median of 2 years 9 months of age (interquartile range: 13 months to 5 years y 10 months). 50% of the cases were associated to chickenpox. Two died. No patient was previously vaccinated against varicella zoster virus. The location was trunk (14), extremities (7) and neck (1). All conscious patients presented cutaneous hyperesthesia. Surgical debridement was performed with a median of 6.8 hours from the onset of hyperesthesia and 2 hours from diagnostic suspicion. The most frequent microorganisms were: Streptococcus pyogenes (38%) and Escherichia coli (31%). The most frequent antibiotics used were penicillin plus clindamycin. The wound was handled with primary closure, grafts and / or flaps. Five patients evolved with sequelae that needed treatment. CONCLUSION: It was identified that STNI in children is associated with chickenpox. It should be suspected in cutaneous lesions and disproportionate hyperesthesia. Aggressive resection is essential for treatment
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Humanos , Masculino , Feminino , Lactente , Pré-Escolar , Infecções dos Tecidos Moles/patologia , Infecções dos Tecidos Moles/terapia , Fasciite Necrosante/patologia , Fasciite Necrosante/terapia , Staphylococcus aureus/isolamento & purificação , Streptococcus pyogenes/isolamento & purificação , Infecções dos Tecidos Moles/cirurgia , Infecções dos Tecidos Moles/microbiologia , Fasciite Necrosante/cirurgia , Fasciite Necrosante/microbiologia , Assistência Perioperatória , Desbridamento , Escherichia coli/isolamento & purificação , Antibacterianos/uso terapêuticoRESUMO
Here, we determined the 5-hydroxymethylcytosine (5hmC), 5-methylcytosine (5mC), Ten Eleven Translocation (TETs), and DNA methyltransferases (DNMTs) levels in epithelial and inflammatory cells of labial salivary glands (LSG) from Sjögren's syndrome (SS)-patients and the effect of cytokines on HSG cells. LSG from SS-patients, controls and HSG cells incubated with cytokines were analysed. Levels of 5mC, 5hmC, DNMTs, TET2 and MeCP2 were assessed by immunofluorescence. In epithelial cells from SS-patients, an increase in TET2, 5hmC and a decrease in 5mC and MeCP2 were observed, additionally, high levels of 5mC and DNMTs and low levels of 5hmC were detected in inflammatory cells. Cytokines increased TET2 and 5hmC and decreased 5mC levels. Considering that the TET2 gene.promoter contains response elements for transcription factors activated by cytokines, together to in vitro results suggest that changes in DNA hydroxymethylation, resulting from altered levels of TET2 are likely to be relevant in the Sjögren's syndrome etiopathogenesis.
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5-Metilcitosina/análogos & derivados , DNA (Citosina-5-)-Metiltransferases/genética , Proteínas de Ligação a DNA/genética , Proteína 2 de Ligação a Metil-CpG/genética , Proteínas Proto-Oncogênicas/genética , Glândulas Salivares Menores/metabolismo , Síndrome de Sjogren/genética , 5-Metilcitosina/metabolismo , Adulto , Idoso , Citidina Desaminase/genética , Citidina Desaminase/metabolismo , Citocinas/imunologia , DNA (Citosina-5-)-Metiltransferase 1/genética , DNA (Citosina-5-)-Metiltransferase 1/metabolismo , DNA (Citosina-5-)-Metiltransferases/metabolismo , Metilação de DNA , DNA Metiltransferase 3A , Proteínas de Ligação a DNA/imunologia , Proteínas de Ligação a DNA/metabolismo , Dioxigenases/genética , Dioxigenases/imunologia , Dioxigenases/metabolismo , Epigênese Genética , Feminino , Expressão Gênica , Regulação da Expressão Gênica , Humanos , Inflamação/genética , Inflamação/metabolismo , Lábio , Masculino , Proteína 2 de Ligação a Metil-CpG/metabolismo , Pessoa de Meia-Idade , Oxigenases de Função Mista/genética , Oxigenases de Função Mista/imunologia , Oxigenases de Função Mista/metabolismo , Regiões Promotoras Genéticas , Proteínas Proto-Oncogênicas/imunologia , Proteínas Proto-Oncogênicas/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Glândulas Salivares Menores/citologia , Glândulas Salivares Menores/imunologia , Síndrome de Sjogren/imunologia , Síndrome de Sjogren/metabolismo , Adulto Jovem , DNA Metiltransferase 3BRESUMO
Objectives: Labial salivary glands (LSGs) of SS patients show alterations related to endoplasmic reticulum stress. Glandular dysfunction could be partly the consequence of an altered inositol-requiring enzyme 1α (IRE1α)/X box-binding protein 1 (XBP-1) signalling pathway of the unfolded protein response, which then regulates genes involved in biogenesis of the secretory machinery. This study aimed to determine the expression, promoter methylation and localization of the IRE1α/XBP-1 pathway components in LSGs of SS patients and also their expression induced by IFN-γ in vitro. Methods: IRE1α, XBP-1 and glucose-regulated protein 78 (GRP78) mRNA and protein levels were measured by qPCR and western blot, respectively, in LSGs of SS patients (n = 47) and control subjects (n = 37). Methylation of promoters was evaluated by methylation-sensitive high resolution melting, localization was analysed by immunofluorescence and induction of the IRE1α/XBP-1 pathway components by IFN-γ was evaluated in 3D acini. Results: A significant decrease of IRE1α, XBP-1u, XBP-1s, total XBP-1 and GRP78 mRNAs was observed in LSGs of SS patients, which was correlated with increased methylation levels of their respective promoters, and consistently the protein levels for IRE1α, XBP-1s and GRP78 were observed to decrease. IFN-γ decreased the mRNA and protein levels of XBP-1s, IRE1α and GRP78, and increased methylation of their promoters. Significant correlations were also found between IRE1α/XBP-1 pathway components and clinical parameters. Conclusion: Decreased mRNA levels for IRE1α, XBP-1 and GRP78 can be partially explained by hypermethylation of their promoters and is consistent with chronic endoplasmic reticulum stress, which may explain the glandular dysfunction observed in LSGs of SS patients. Additionally, glandular stress signals, including IFN-γ, could modulate the expression of the IRE1α/XBP-1 pathway components.
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Endorribonucleases/genética , Regulação da Expressão Gênica , Proteínas Serina-Treonina Quinases/genética , RNA Mensageiro/genética , Glândulas Salivares/fisiopatologia , Síndrome de Sjogren/genética , Proteína 1 de Ligação a X-Box/genética , Adulto , Idoso , Western Blotting , Metilação de DNA , Chaperona BiP do Retículo Endoplasmático , Estresse do Retículo Endoplasmático , Endorribonucleases/biossíntese , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Proteínas Serina-Treonina Quinases/biossíntese , Glândulas Salivares/metabolismo , Transdução de Sinais , Síndrome de Sjogren/metabolismo , Síndrome de Sjogren/patologia , Proteína 1 de Ligação a X-Box/biossíntese , Adulto JovemRESUMO
BACKGROUND: The effect of food fortification with folic acid on the incidence of lip-palate fissures (FLP) is under discussion. AIM: To calculate the rate of hospital discharges due to cleft lip and palate (CLP) and explore whether they decreased after the start of folic acid fortification in Chile. MATERIAL AND METHODS: The hospital discharge databases published by the Chilean Ministry of Health were analyzed. The trends of discharge rates due to CLP of children of less than one year of life (ICD-10 Q350-379 code) and live births (lb) were calculated from 1986 to 2012. Variables such as incidence of rubella, poverty rate, obesity in pregnant women, and percentage of women who smoked or drank alcohol in the last month were included. The relative risk (RR) of CLP pre-fortification (1986-1999) and post-fortification (2000-2010) was calculated. Mixed and Prais-Winsten models were used to adjust other variables. RESULTS: The hospital discharge rate due to CLP decreased from 1.88 x 1,000 lb during 1986-1996 to 1.68 x 1,000 lb (RR = 1.09 (95% CI 1.05-1.14, p < 0.001). Prais-Winsten and Mixed models, determined that folic acid fortification is the variable explaining this decrease. During 2004-2011, hospital discharge rates for cleft lip decreased but there was an increase in the figures for cleft lip and palate. CONCLUSIONS: The 9% decrease in hospital discharge rates for CLP is concomitant with wheat folic acid fortification.