RESUMEN
Trueperella pyogenes (T. pyogenes) is a common opportunistic pathogen of many livestock and play an important regulation role during multibacterial infection and interaction with the host by its primary virulence factor pyolysin (PLO). The purpose of this study was to investigate the regulation role of PLO which serve as a combinational pathogen with lipopolysaccharide (LPS) during endometritis. In this study, the expression of bioactive recombinant PLO (rPLO) in a prokaryotic expression system and its purification are described. Moreover, we observed that rPLO inhibited the innate immune response triggered by LPS and that methyl-ß-cyclodextrin (MBCD) abrogated this inhibitory effect in goat endometrium stromal cells (gESCs). Additionally, we show from pharmacological and genetic studies that rPLO-induced autophagy represses gene expression by inhibiting NLRP3 inflammasome activation. Importantly, this study reported that ATF6 serves as a primary regulator of the cellular inflammatory reaction to rPLO. Overall, these observations suggest that T. pyogenes PLO could create an immunosuppressive environment for other pathogens invasion by regulating cellular signaling pathways.
Asunto(s)
Factor de Transcripción Activador 6/metabolismo , Autofagia/efectos de los fármacos , Proteínas Bacterianas/farmacología , Toxinas Bacterianas/farmacología , Endometrio/citología , Proteínas Hemolisinas/farmacología , Lipopolisacáridos/farmacología , Actinomycetaceae/patogenicidad , Animales , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Toxinas Bacterianas/genética , Toxinas Bacterianas/metabolismo , Femenino , Cabras , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Inflamasomas/efectos de los fármacos , Inflamasomas/metabolismo , Inflamación , Mediadores de Inflamación/metabolismo , Proteína con Dominio Pirina 3 de la Familia NLR/metabolismo , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacología , Células del Estroma/efectos de los fármacos , Células del Estroma/metabolismoRESUMEN
Objective. To identify reproductive health barriers and perceptions regarding family planning among mothers in ten rural communities of Guatemala. Methods. Data were collected from 85 women in a Nutrition Recuperation Project (NRP) conducted by a freestanding nonprofit clinic in Palajunoj Valley, Guatemala. All nonpregnant women participating in the NRP were eligible for enrollment in this study, and NRP staff members aided in their enrollment. Participants were interviewed and data were entered into a structured questionnaire. Data analysis was conducted using R version 1.1.456. Results. After asking participants if they believed fertility is higher on certain days, only 5 women (5.9%) correctly identified these days as occurring in the middle of the menstrual cycle. 35 women (41.2%) practiced some form of family planning, and 27 (31.8%) reported that they do not know of a place where they could obtain a contraceptive method. Conclusion. There is a lack of education regarding family planning methods in this valley, and the levels of contraception use are below average for rural Guatemala. These findings may implicate substantial health risks for women and children in the valley, and they support the pertinence of education-based interventions in the area of reproductive health behaviors.
RESUMEN
OBJECTIVE: To study the effect of Lactobacillus sp. A-2 metabolites on viability of CAL-27 cells and apoptosis in CAL-27 cells. METHODS: Lactobacillus sp. A-2 metabolites 1 and 2 (LM1 and LM2) were obtained by culturing Lactobacillus sp. A-2 in reconstituted whey medium and whey-inulin medium; the cultured CAL-27 cells were treated with different concentrations of LM1 and LM2 (0, 3, 6, 12, 24, 48 mg/mL) and assayed by methyl thiazolyltetrazolium (MTT) method; morphological changes of apoptotic cell were observed under fluorescence microscopy by acridine orange (Ao) fluorescent staining; flow cytometry method (FCM) and agarose gel electrophoresis were used to detect the apoptosis of CAL-27 cells treated LM1 and LM2. RESULTS: The different concentrations of LM1 and LM2 could restrain the growth of CAL-27 cells, and in a dose-dependent manner; the apoptosis of CAL-27 cells was obviously induced and was time-dependent. CONCLUSIONS: Viability of CAL-27 cells was inhibited by Lactobacillus sp. A-2 metabolites; Lactobacillus sp. A-2 metabolites could induce CAL-27 cells apoptosis; study on the bioactive compounds in the Lactobacillus sp. A-2 metabolites and their molecular mechanism is in progress.
Asunto(s)
Apoptosis/fisiología , Carcinoma de Células Escamosas/patología , Lactobacillus/metabolismo , Neoplasias de la Lengua/patología , Análisis de Varianza , Supervivencia Celular , Células Cultivadas , Electroforesis en Gel de Agar , Citometría de Flujo , Humanos , Lactobacillus/crecimiento & desarrollo , Microscopía Fluorescente , Sales de Tetrazolio , Tiazoles , Factores de TiempoRESUMEN
Objective: To study the effect of Lactobacillus sp. A-2 metabolites on viability of CAL-27 cells and apoptosis in CAL-27 cells. Methods: Lactobacillus sp. A-2 metabolites 1 and 2 (LM1 and LM2) were obtained by culturing Lactobacillus sp. A-2 in reconstituted whey medium and whey-inulin medium; the cultured CAL-27 cells were treated with different concentrations of LM1 and LM2 (0, 3, 6, 12, 24, 48 mg/mL) and assayed by methyl thiazolyltetrazolium (MTT) method; morphological changes of apoptotic cell were observed under fluorescence microscopy by acridine orange (Ao) fluorescent staining; flow cytometry method (FCM) and agarose gel electrophoresis were used to detect the apoptosis of CAL-27 cells treated LM1 and LM2. Results: The different concentrations of LM1 and LM2 could restrain the growth of CAL-27 cells, and in a dose-dependent manner; the apoptosis of CAL-27 cells was obviously induced and was time-dependent. Conclusions: Viability of CAL-27 cells was inhibited by Lactobacillus sp. A-2 metabolites; Lactobacillus sp. A-2 metabolites could induce CAL-27 cells apoptosis; study on the bioactive compounds in the Lactobacillus sp. A-2 metabolites and their molecular mechanism is in progress. .