RESUMO
The adult mammalian cardiomyocyte has a very limited capacity to reenter the cell cycle and advance into mitosis. Therefore, diseases characterized by lost contractile tissue usually evolve into myocardial remodeling and heart failure. Analyzing the cardiac transcriptome at different developmental stages in a large mammal closer to the human than laboratory rodents may serve to disclose positive and negative cardiomyocyte cell cycle regulators potentially targetable to induce cardiac regeneration in the clinical setting. Thus we aimed at characterizing the transcriptomic profiles of the early fetal, late fetal, and adult sheep heart by employing RNA-seq technique and bioinformatic analysis to detect protein-encoding genes that in some of the stages were turned off, turned on, or differentially expressed. Genes earlier proposed as positive cell cycle regulators such as cyclin A, cdk2, meis2, meis3, and PCNA showed higher expression in fetal hearts and lower in AH, as expected. In contrast, genes previously proposed as cell cycle inhibitors, such as meis1, p16, and sav1, tended to be higher in fetal than in adult hearts, suggesting that these genes are involved in cell processes other than cell cycle regulation. Additionally, we described Gene Ontology (GO) enrichment of different sets of genes. GO analysis revealed that differentially expressed gene sets were mainly associated with metabolic and cellular processes. The cell cycle-related genes fam64a, cdc20, and cdk1, and the metabolism-related genes pitx and adipoq showed strong differential expression between fetal and adult hearts, thus being potent candidates to be targeted in human cardiac regeneration strategies.NEW & NOTEWORTHY We characterized the transcriptomic profiles of the fetal and adult sheep hearts employing RNAseq technique and bioinformatic analyses to provide sets of transcripts whose variation in expression level may link them to a specific role in cell cycle regulation. It is important to remark that this study was performed in a large mammal closer to humans than laboratory rodents. In consequence, the results can be used for further translational studies in cardiac regeneration.
Assuntos
Regulação da Expressão Gênica no Desenvolvimento , Coração/fisiologia , Miocárdio/metabolismo , Regeneração , Transcriptoma , Animais , Ciclina A/genética , Ciclina A/metabolismo , Feminino , Coração/crescimento & desenvolvimento , Masculino , Ovinos , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
BACKGROUND: In Argentina, there has been an abrupt increase in KPC-2-producing Klebsiella pneumoniae (K. pneumoniae). Tucumán is a multi-border area, so the rapid dissemination of carbapenem-resistant K. pneumoniae is a clinically relevant problem for the region. OBJECTIVES: This study aimed to investigate the epidemiological and molecular patterns of KPC-producing K. pneumoniae clinical isolates collected from different hospitals in Tucumán. METHODS: Carbapenem-resistant K. pneumoniae strains were sequentially and uniquely collected during two time periods. Antibiotic susceptibility was determined by the automated Vitex 2® system and using the standard agar dilution test. Multilocus sequence typing and pulsed-field electrophoresis were used for epidemiological analysis. The genetic structures around blaKPC and the encoding genes of extended-spectrum ß-lactamases were detected by polymerase chain reaction and sequencing. Plasmids were analysed by conjugation and using the plasmid relaxase gene-typing method. RESULTS: All 37 isolates were multidrug resistant, and theblaKPC-2 gene was confirmed in all of them. In 17 isolates (45.9%), the blaCTX-M-2 gene was also amplified, as well as blaSHV-2 in five isolates (13.5%) and blaCTX-M-2/blaSHV-2 in four isolates (10.8%). The molecular epidemiology of the blaKPC-2 gene has resulted in it being associated with an IncL/M transferable plasmid disseminating in various sequence types (STs) (ST17, ST556, ST342, ST147, ST461, ST65, ST15 and ST70), and in a new genetic environment with a 764-bp deletion in the ISKpn7-blaKPC region. CONCLUSIONS: These findings contribute to the understanding of the great diversity of the blaKPC-2-carrying genetic platforms.
Assuntos
Carbapenêmicos/farmacologia , Infecções por Klebsiella/microbiologia , Klebsiella pneumoniae/genética , beta-Lactamases/metabolismo , Argentina , Farmacorresistência Bacteriana , Eletroforese em Gel de Campo Pulsado , Humanos , Infecções por Klebsiella/tratamento farmacológico , Klebsiella pneumoniae/metabolismo , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Tipagem de Sequências Multilocus , Plasmídeos/genética , Estudos Retrospectivos , beta-Lactamases/genéticaRESUMO
Los bovinos son el principal reservorio de Escherichia coli productor de toxina Shiga (STEC); las estrategias para evitar su transmisión se concentran en la planta de faena. El objetivo de este trabajo fue evaluar la calidad higiénico-sanitaria y la frecuencia de detección de STEC en medias reses bovinas de frigoríficos de tránsito provincial. Se procesaron 274 esponjados de media res; en 9 (3,3%) el recuento de E. coli genérico fue marginal, en 4 (1,4%) se aisló E. coli O157, de los cuales 2 fueron caracterizados como stx2c(vh-a)/eae/ehxA, y los otros 2 como no toxigénicos. A partir de una (0,4%) muestra se aisló E. coli no-O157 ONT:H49, stx2a/ehxA/saa. En este trabajo la calidad del producto analizado indica que en la provincia de Tucumán se cumplen las buenas prácticas de manufactura en la faena de bovinos.
Cattle are the main reservoir of Shiga toxin-producing Escherichia coli (STEC), and the strategies to prevent the transmission of these microorganisms are concentrated in the slaughtering plant. The aim of this study was to evaluate the hygienic-sanitary quality and the frequency of detection of STEC in beef carcasses in abattoirs from Tucuman province. Two hundred and seventy four beef carcass sponges were processed; the count of generic E. coli was marginal in 9 (3,3%) of them. Escherichia coli O157 was isolated in 4 (1,4%) samples; 2 of which were characterized as stx2c(vh-a)/eae/ehxA whereas the other 2 were non-toxigenic strains. Non-O157 E. coli ONT:H49, stx2a/ehxA/saa was isolated from 1 sample (0,4%). In this work the quality of the analyzed product indicates that the good practices of manufacture are fulfilled in slaughtering facilities in Tucumán province.
Assuntos
Animais , Bovinos , Matadouros , Escherichia coli Shiga Toxigênica , Carne , Argentina , Escherichia coli O157 , Proteínas de Escherichia coli/análise , Infecções por Escherichia coli , Escherichia coli Shiga Toxigênica/isolamento & purificação , Carne/microbiologiaRESUMO
Cattle are the main reservoir of Shiga toxin-producing Escherichia coli (STEC), and the strategies to prevent the transmission of these microorganisms are concentrated in the slaughtering plant. The aim of this study was to evaluate the hygienic-sanitary quality and the frequency of detection of STEC in beef carcasses in abattoirs from Tucuman province. Two hundred and seventy four beef carcass sponges were processed; the count of generic E. coli was marginal in 9 (3,3%) of them. Escherichia coli O157 was isolated in 4 (1,4%) samples; 2 of which were characterized as stx2c(vh-a)/eae/ehxA whereas the other 2 were non-toxigenic strains. Non-O157 E. coli ONT:H49, stx2a/ehxA/saa was isolated from 1 sample (0,4%). In this work the quality of the analyzed product indicates that the good practices of manufacture are fulfilled in slaughtering facilities in Tucumán province.
Assuntos
Matadouros , Carne , Escherichia coli Shiga Toxigênica , Animais , Argentina , Bovinos , Infecções por Escherichia coli , Escherichia coli O157 , Proteínas de Escherichia coli/análise , Carne/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificaçãoRESUMO
Escherichia coli O157 is an emergent pathogen associated with diarrhea, hemorrhagic colitis and hemolytic uremic syndrome. Meat products constitute an important transmission source of this microorganism. The aims of this study were to characterize E. coli O157 isolated from cattle and meat products collected from abattoirs and retail stores, to establish the clonal relatedness among regional isolates and to compare them with those in the national database. Between 2004 and 2013, 169 minced meat, 35 sausage and 216 carcass samples were analyzed. Thirteen E. coli O157 isolates were identified; 6 of which were O157:H7 and characterized as stx2c(vh-a)/eae/ehxA (n = 5) and stx2/eae/ehxA (n = 1). The 7 remaining isolates were non-toxigenic E. coli strains, and serotyped as O157:NT (n = 4), O157:NM (n = 1), O157:ND (n = 1) and O157:H16 (n = 1). The strains yielded different XbaI-PFGE patterns. Compared to the E. coli O157 isolates in the National Database, none of these patterns have been previously detected in strains of different origin in Argentina.
Assuntos
Bovinos/microbiologia , Escherichia coli O157/isolamento & purificação , Microbiologia de Alimentos , Carne/microbiologia , Animais , Argentina , Técnicas de Tipagem Bacteriana , DNA Bacteriano/genética , Reservatórios de Doenças , Eletroforese em Gel de Campo Pulsado , Infecções por Escherichia coli/transmissão , Escherichia coli O157/classificação , Escherichia coli O157/genética , Escherichia coli O157/patogenicidade , Genes Bacterianos , Produtos da Carne/microbiologia , Polimorfismo de Fragmento de Restrição , Sorotipagem , Virulência/genéticaRESUMO
Escherichia coli O157 es un patógeno emergente asociado a diarrea, colitis hemorrágica y síndrome urémico hemolítico. Los productos cárnicos constituyen una importante fuente de contaminación con este microorganismo. Los objetivos de este estudio fueron establecer la frecuencia de detección de E. coli O157 en productos cárnicos y media res en la provincia de Tucumán, caracterizar los factores de virulencia de los aislamientos obtenidos, establecer la relación clonal entre cepas regionales mediante electroforesis de campo pulsado y comparar con lo consignado en la base de datos nacional. Desde 2004 hasta 2013 se analizaron 169 muestras de carne picada, 35 embutidos y 216 esponjados de media res. Se identificaron 13 aislamientos de E. coli O157; 6 de ellos fueron O157:H7 productores de toxina Shiga y se caracterizaron como stx2c(vh-a)/eae/ehxA (n = 5) y stx2/eae/ehxA (n = 1); los 7 aislamientos de E. coli O157 no toxigénicos fueron O157:NT(n = 4),O157:NM (n = 1),O157:ND (n = 1) y O157:H16 (n = 1). Los patrones de PFGE fueron diferentes entre sí y de los registrados en la base de datos nacional. Se concluye que existe gran diversidad genética en los aislamientos de E. coli O157 circulantes en nuestra región
Escherichia coli O157 is an emergent pathogen associated with diarrhea, hemorrhagic colitis and hemolytic uremic syndrome. Meat products constitute an important transmission source of this microorganism. The aims of this study were to characterize E. coli O157 isolated from cattle and meat products collected from abattoirs and retail stores, to establish the clonal relatedness among regional isolates and to compare them with those in the national database. Between 2004 and 2013, 169 minced meat, 35 sausage and 216 carcass samples were analyzed. Thirteen E. coli O157 isolates were identified; 6 of which were O157:H7 and characterized as stx2c(vh-a)/eae/ehxA (n = 5) and stx2/eae/ehxA (n = 1). The 7 remaining isolates were non-toxigenic E. coli strains, and serotyped as O157:NT (n = 4), O157:NM (n = 1), O157:ND (n = 1) and O157:H16 (n = 1). The strains yielded different XbaI-PFGE patterns. Compared to the E. coli O157 isolates in the National Database, none of these patterns have been previously detected in strains of different origin in Argentina
Assuntos
Animais , Bovinos , Escherichia coli O157/isolamento & purificação , Escherichia coli O157/genética , Produtos da Carne/análise , Bases de Dados Bibliográficas/estatística & dados numéricos , Eletroforese em Gel de Campo Pulsado/métodos , Escherichia coli O157/classificação , Fatores de Virulência/análiseRESUMO
En equipamientos industriales la persistencia de las bacterias se debe al desarrollo de biofilms. El hipoclorito de sodio (NaClO) es usado rutinariamente como desinfectante. El objetivo del trabajo fue determinar el efecto inhibitorio del NaClO sobre S. cohnii productoras y no productoras de biofilms aisladas de una fabrica de pastas a partir de maquinarias de un establecimiento de Tucuman, Argentina. Se estudio produccion de biofilm, concentracion inhibitoria minima (CIM), concentracion bactericida minima (CBM) y curva de tiempo de muerte con NaClO frente a S. cohnii productora y no productora de biofilm. Los resultados obtenidos demostraron que de los ocho cocos grampositivos aislados, cuatro correspondieron a S. cohnii. Los valores CIM y CBM para S. cohnii productoras y no productoras de biofilm resultaron entre 0,05 y 0,2 g/dL. Mediante la curva de muerte se determino efecto bactericida observando una disminucion de ≥4 log de ufc (1h) tratado con 0,2 y 0,4% de NaClO. La formacion de biofilm para la cepa no tratada fue de DO: 0,12 y para la cepa tratada DO: 0,07 a las 18 h. Aplicar desinfectantes con amplio espectro de accion es importante, debido a que la eliminacion de bacterias puede prevenir su diseminacion y evitar la formacion de biofilms.(AU)
Persistence of bacteria in industrial equipment is favoured by biofilm formation. Sodium hypochlorite (NaClO) is routinely used as a disinfectant. The objective of the current study was to determine the inhibitory effect of NaClO on biofilm-producing and non-biofilm-producing S. cohnii strains, isolated from equipment in a fresh pasta factory in Tucuman, Argentina. The following parameters were assayed: production of biofilm, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and NaClO death curves for both biofilm-producing and non-biofilm-producing S. cohnii strains. Four of the eight isolated Gram-positive cocci corresponded to S. cohnii. MIC and MBC values for biofilm-producing and non-biofilm-producing S. cohnii strains were between 0.05 and 0.2 g/dL. A bactericidal effect of 0.2 and 0.4% NaClO for 1 h was established with the death curve, showing a decrease in cfu of ≥4 log units. Biofilm production after 18 h for untreated strains gave an OD of 0.12, whereas the OD of treated strains was 0.07. The use of broad spectrum disinfectants is important, because elimination of bacteria helps prevent their propagation and avoid the formation of biofilms.(AU)
A persistÛncia da bactéria em equipamento industrial e resultado do desenvolvimento de biofilmes. O hipoclorito de sodio (NaOCl) e rotineiramente utilizado como desinfetante. O objetivo do trabalho foi determinar o efeito inibitorio do NaClO sobre S. cohnii produtoras e nao produtoras de biofilmes isoladas de uma fabrica de massas frescas a partir de maquinarias de um estabelecimento de Tucuman, Argentina. Foram estudados: producao de biofilme, concentracao inibitoria minima (CIM), concentracao bactericida minima (CBM) e curva de tempo de morte com NaClO perante S. cohnii produtora e naoprodutora de biofilme. Os resultados obtidos demonstraram que quatro dos oito cocos Gram-positivos isolados corresponderam a S. cohnii. Os valores CIM e CBM para S. cohnii produtoras e nao produtoras de biofilme foram entre 0,05 e 0,2 g/dL. Atraves da curva de morte foi determinado um efeito bactericida observando uma diminuicao de ≥ 4 log de ufc (1h) tratado com 0,2 e 0,4% de NaClO. A formacao de biofilme, apos 18 h, para a cepa nao tratada foi de DO: 0,12, ao passo que para a cepa tratada foi de 0,07. A utilizacao de desinfetantes de amplo espectro de acao e importante, visto que a eliminaþÒo de bacterias pode prevenir sua propagaþÒo e evitar a formacao de biofilmes.(AU)
RESUMO
En equipamientos industriales la persistencia de las bacterias se debe al desarrollo de biofilms. El hipoclorito de sodio (NaClO) es usado rutinariamente como desinfectante. El objetivo del trabajo fue determinar el efecto inhibitorio del NaClO sobre S. cohnii productoras y no productoras de biofilms aisladas de una fabrica de pastas a partir de maquinarias de un establecimiento de Tucuman, Argentina. Se estudio produccion de biofilm, concentracion inhibitoria minima (CIM), concentracion bactericida minima (CBM) y curva de tiempo de muerte con NaClO frente a S. cohnii productora y no productora de biofilm. Los resultados obtenidos demostraron que de los ocho cocos grampositivos aislados, cuatro correspondieron a S. cohnii. Los valores CIM y CBM para S. cohnii productoras y no productoras de biofilm resultaron entre 0,05 y 0,2 g/dL. Mediante la curva de muerte se determino efecto bactericida observando una disminucion de ≥4 log de ufc (1h) tratado con 0,2 y 0,4% de NaClO. La formacion de biofilm para la cepa no tratada fue de DO: 0,12 y para la cepa tratada DO: 0,07 a las 18 h. Aplicar desinfectantes con amplio espectro de accion es importante, debido a que la eliminacion de bacterias puede prevenir su diseminacion y evitar la formacion de biofilms.
Persistence of bacteria in industrial equipment is favoured by biofilm formation. Sodium hypochlorite (NaClO) is routinely used as a disinfectant. The objective of the current study was to determine the inhibitory effect of NaClO on biofilm-producing and non-biofilm-producing S. cohnii strains, isolated from equipment in a fresh pasta factory in Tucuman, Argentina. The following parameters were assayed: production of biofilm, minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and NaClO death curves for both biofilm-producing and non-biofilm-producing S. cohnii strains. Four of the eight isolated Gram-positive cocci corresponded to S. cohnii. MIC and MBC values for biofilm-producing and non-biofilm-producing S. cohnii strains were between 0.05 and 0.2 g/dL. A bactericidal effect of 0.2 and 0.4% NaClO for 1 h was established with the death curve, showing a decrease in cfu of ≥4 log units. Biofilm production after 18 h for untreated strains gave an OD of 0.12, whereas the OD of treated strains was 0.07. The use of broad spectrum disinfectants is important, because elimination of bacteria helps prevent their propagation and avoid the formation of biofilms.
A persistência da bactéria em equipamento industrial e resultado do desenvolvimento de biofilmes. O hipoclorito de sodio (NaOCl) e rotineiramente utilizado como desinfetante. O objetivo do trabalho foi determinar o efeito inibitorio do NaClO sobre S. cohnii produtoras e nao produtoras de biofilmes isoladas de uma fabrica de massas frescas a partir de maquinarias de um estabelecimento de Tucuman, Argentina. Foram estudados: producao de biofilme, concentracao inibitoria minima (CIM), concentracao bactericida minima (CBM) e curva de tempo de morte com NaClO perante S. cohnii produtora e naoprodutora de biofilme. Os resultados obtidos demonstraram que quatro dos oito cocos Gram-positivos isolados corresponderam a S. cohnii. Os valores CIM e CBM para S. cohnii produtoras e nao produtoras de biofilme foram entre 0,05 e 0,2 g/dL. Atraves da curva de morte foi determinado um efeito bactericida observando uma diminuicao de ≥ 4 log de ufc (1h) tratado com 0,2 e 0,4% de NaClO. A formacao de biofilme, apos 18 h, para a cepa nao tratada foi de DO: 0,12, ao passo que para a cepa tratada foi de 0,07. A utilizacao de desinfetantes de amplo espectro de acao e importante, visto que a eliminação de bacterias pode prevenir sua propagação e evitar a formacao de biofilmes.
Assuntos
Biofilmes , Hipoclorito de Sódio/toxicidade , Staphylococcus/fisiologia , Argentina , Indústria Alimentícia , Indústrias , Massas AlimentíciasRESUMO
In the last years, Enterobacteriaceae such as Klebsiella pneumoniae, Proteus mirabilis and Escherichia coli, have acquired resistance to third-generation cephalosporins (C3G) because of the presence of plasmid-mediated AmpC ß-lactamases. The aim of this work was to detect plasmid AmpC enzymes and to investigate the predominant types in our region. Between March and July 2009, 733 consecutive isolates of Enterobacteriaceae derived from hospitals and outpatient centers were studied. Susceptibility testing was performed by disk diffusion; one P. mirabilis and three E. coli strains showed resistance to cephamycins (cefoxitin) and C3G. An E-test to determine MIC and a synergy test by aminophenylboronic disks were performed. Enzymatic activity against cefoxitin was confirmed by a microbiological assay. A polymerase chain reaction (PCR) for the detection of plasmid-mediated ampC genes of different groups was performed and a 462-bp amplicon was obtained when using primers directed against the CIT group; the obtained sequences were compared to blaCMY-2 sequences, showing 100% identity. The emergence of CMY-2-type plasmid-mediated AmpC ß-lactamases indicated the importance of implementing systematic monitoring of these resistances to avoid potential clinical and epidemiological consequences.