Identificación de enterobacterias multirresistentes a antibióticos en muestras de heces de lactantes residentes en Talara, Piura, Perú / Identification of multidrug-resistant enterobacteriaceae in fecal samples from infants residing in Talara, Piura, Peru Arturo

La colonización fecal en lactantes por bacterias resistentes a los antimicrobianos es un potencial riesgo para futuras terapias antibióticas. Nuestro objetivo fue determinar la frecuencia y características sociodemográficas de lactantes portadores fecales de enterobacterias resistentes a ciprofloxacina (PFRC) y sus genes de resistencia asociados. Analizamos muestras fecales de 41 niños lactantes residentes en el distrito de Talara-Piura, Perú, en 2019. Evaluamos la presencia de 3 genes de resistencia a quinolonas: aac(6')-Ib-cr, qnrB y oqxA y 2 de betalactamasas: bla CTX-M, bla PER-2.El 68% de lactantes fueron PFRC, Escherichia coli (83,3%) fue el más frecuente. El análisis genotípico detectó: oqxA (41,1%), qnrB (26,7%) y aac(6')-Ib-cr (20%) y al gen bla CTX-M en el 93,3% de los aislados con betalactamasas. La elevada frecuencia de PFRC nos alertan sobre el potencial riesgo en la pérdida de utilidad de esta familia antibiótica en el área de estudio.

Fecal colonization by antimicrobial-resistant bacteria in infants is a potential risk for future antibiotic therapy. We aimed to determine the sociodemographic characteristics and frequency of infants who were fecal carriers of ciprofloxacin-resistant enterobacteriaceae (FCCRE) and their associated resistance genes. We analyzed fecal samples from 41 infants from the district of Talara, Piura, Peru in 2019. The presence of 3 quinolone resistance genes was evaluated: aac(6')-Ib-cr, qnrB and oqxA as well as of 2 beta-lactamase genes: bla CTX-M,bla PER-2. We found that 68% of infants were FCCRE, Escherichia coli (83.3%) was the most frequent bacteria. The genotypic analysis detected: oqxA (41.1%), qnrB (26.7%), aac(6')-Ib-cr (20%) and the bla CTX-M gene (93.3%) of the isolates with beta-lactamases. The high frequency of FCCRE alerts us of the potential risk of this antibiotic family becoming less useful over time.

Infección de piel y tejidos blandos por Burkholderia stabilis en un niño con quemaduras graves / Skin and soft tissue infection due to Burkholderia stabilis in a severely burned child

Resumen Las heridas por quemadura representan un grave problema, sobre todo en la población pediátrica, dada la severidad de su presentación y la morbimortalidad asociada. La infección es la complicación más frecuente y grave en el paciente quemado. Las bacterias que conforman el complejo Burkholderia cepacia (CBc) son capaces de causar enfermedades en plantas, humanos y animales. En el hombre pueden establecer infecciones crónicas y frecuentemente graves, por lo general en pacientes con fibrosis quística y en inmunocomprometidos. El CBc está compuesto por al menos 22 especies filogenéticamente muy relacionadas. El objetivo de esta publicación fue describir el primer caso de una infección de piel y partes blandas por Burkholderia stabilis, una especie poco frecuente, en un niño con grandes quemaduras en la Argentina. Las especies del CBc son intrínsecamente resistentes a la mayoría de los antimicrobianos disponibles clínicamente, como aminoglucósidos, quinolonas, polimixinas y β-lactámicos. Esto representa un serio problema en el momento de tratar las infecciones por las escasas opciones terapéuticas.

Abstract Burn wounds represent a serious problem, especially in the pediatric population, given the severity of their presentation and the associated morbidity and mortality. Infection is the most frequent and serious complication in the burned patient. Burkholderia cepacia (CBc) complex bacteria are capable of causing disease in plants, humans, and animals. In human beings they can establish chronic and frequently serious infections, generally in patients with cystic fibrosis and in immunocompromised patients. The CBc is composed of 22 phylogenetically closely related species. The objective of this publication was to describe the first report of a skin and soft tissue infection by Burkholderia stabilis, a rare species, in a child with extensive burns in Argentina. CBc species are inherently resistant to most clinically available antimicrobials, such as aminoglycosides, quinolones, polymyxins, and β-lactams. This represents a serious problem when treating infections, due to the limited therapeutic options.

Resumo As feridas por queimadura representam um grave problema, principalmente na população pediátrica, devido à gravidade de sua apresentação e morbimortalidade associada. A infecção é a complicação mais frequente e grave do paciente queimado. As bactérias que compõem o complexo Burkholderia cepacia (CBc) são capazes de causar doenças em plantas, humanos e animais. No homem, podem estabelecer infecções crônicas e freqüentemente graves, geralmente em pacientes com fibrose cística e imunocomprometidos. O CBc é composto, no mínimo, por 22 espécies filogeneticamente muito relacionadas. O objetivo desta publicação é descrever o primeiro caso de uma infecção de pele e tecidos moles por Burkholderia stabilis, uma espécie rara, em uma criança com queimaduras extensas na Argentina. As espécies do CBc são inerentemente resistentes à maioria dos antimicrobianos disponíveis clinicamente, como aminoglicosídeos, quinolonas, polimixinas e β-lactâmicos. Isso representa um problema sério na hora de tratar as infecções devido às opções terapêuticas limitadas.

Switching from efavirenz to elvitegravir/cobicistat/emtricitabine/tenofovir alafenamide reduces central nervous system symptoms in people living with HIV / 中华医学杂志(英文版)

BACKGROUND@#Central nervous system (CNS) symptoms after efavirenz (EFV) treatment in people living with human immunodeficiency virus (HIV) could persist and impact their quality of life. We assessed the impact of EFV-based regimen replacement with elvitegravir/cobicistat/emtricitabine/tenofovir alafenamide (E/C/F/TAF), which is considered an alternative option for subjects who do not tolerate EFV. Most specifically, we assessed the safety and the efficacy of E/C/F/TAF and its effects on the participants' neuropsychiatric toxicity symptoms in a real-life setting.@*METHODS@#A prospective cohort study was conducted among virologic suppressed HIV-positive participants receiving EFV-based regimens with ongoing CNS toxicity ≥ grade 2. The participants were switched to single-pill combination regimens E/C/F/TAF and followed up for 48 weeks. The neuropsychiatric toxicity symptoms were measured using a CNS side effects questionnaire, as well as the Hospital Anxiety and Depression Scale and the Pittsburgh Sleep Quality Index. The primary outcome measure was the proportion of participants experiencing grade 2 or higher CNS toxicity after EFV switch off at weeks 12, 24, and 48. Secondary endpoints included virologic and immunological responses and the effect on fasting lipids at week 48 after switch.@*RESULTS@#One hundred ninety-six participants (96.9% men, median age: 37.5 years, median: 3.7 years on prior EFV-containing regimens) were included in the study. Significant improvements in anxiety and sleep disturbance symptoms were observed at 12, 24, and 48 weeks after switching to E/C/F/TAF (P < 0.05). No significant change in depression symptom scores was observed. At 48 weeks after switch, HIV viral load <50 copies/mL was maintained in all of the participants, median fasting lipid levels were moderately increased (total cholesterol [TC]: 8.2 mg/dL, low-density lipoprotein cholesterol [LDL-C]: 8.5 mg/dL, high-density lipoprotein cholesterol [HDL-C]: 2.9 mg/dL, and triglyceride (TG): 1.6 mg/dL, and the TC:HDL-C ratio remained stable.@*CONCLUSIONS@#The single-pill combination regimens E/C/F/TAF is safe and well tolerated. This study reveals that switching from EFV to E/C/F/TAF significantly reduces neuropsychiatric toxicity symptoms in people living with HIV with grade 2 or higher CNS complaints.

Effect of rebamipide on the acute gouty arthritis in rats induced by monosodium urate crystals / 北京大学学报(医学版)

OBJECTIVE@#To investigate the role of rebamipide in the treatment of acute gout arthritis rats induced by monosodium urate (MSU) crystal.@*METHODS@#Forty-two male rats were randomly divided into three groups (n=14). Group A was treated with oral rebamipide, group B with oral colchicine, and group C with oral placebo. The rats were monitored for the induction of arthritis with clinical manifestations and pathological changes, and the levels of interleukin (IL)-1β、IL-6、IL-10, and tumor necrosis factor (TNF)-α in serum were measured.@*RESULTS@#In group C, the clinical score and swelling index reached the maximum in 24 h, and then gradually decreased to 72 h. After 24 h of model induced, the clinical scores in group C were significantly higher than those in group A and group B [2 (1-3) vs. 0 (0-1) vs. 1 (0-2), P < 0.01], the swelling indexes in group C were significantly higher than those in group A and group B [0.36 (0.16-0.52) vs. 0.11 (0-0.20) vs. 0.12 (0-0.16), P < 0.01]. Histologically, after 24 h of model induced, there was a large number of neutrophil infiltration in the synovium of group C [scale score: 4 (2-4)], and there was no significant inflammatory cell infiltration in group A [1 (0-2)] and group B [1 (0-2)], the difference was statistically significant (P < 0.001). After 24 h of model induced, the levels of IL-1β, IL-6, IL-10, and TNF-α in serum of group C were significantly higher than those in group A and B [IL-1β: (41.86±5.72) vs. (27.35±7.47) vs. (27.76±5.28) ng/L, IL-6: (1 575.55±167.11) vs. (963.53±90.22) vs. (964.08±99.31) ng/L, IL-10: (37.96±3.76) vs. (21.68±4.83) vs. (16.20±2.49) ng/L, TNF-α: (21.32±1.34) vs. (15.82±2.54) vs. (17.35±7.47) μg/L, P < 0.001].@*CONCLUSION@#Rebamipide has a protective effect on acute gout arthritis rats induced by MUS crystals.

Resistoma y genómica comparativa de aislados clínicos de Escherichia coli diarreogénica en Lima, Perú / Resistome and comparative genomics of clinical isolates of diarrheagenic Escherichia coli from Lima, Peru

RESUMEN Con el objetivo de describir las características genómicas relacionadas con la resistencia antimicrobiana y genómica comparativa de Escherichia coli diarreogénica (DEC), se sometieron a secuenciamiento genómico catorce aislamientos de DEC del banco de cepas del Instituto Nacional de Salud (INS). Las secuencias obtenidas se analizaron mediante procedimientos bioinformáticos a fin de buscar genes de resistencia microbiana y regiones genéticas relacionadas con patotipos y filogrupos. Se detectaron diversos determinantes de resistencia antimicrobiana, se destaca la producción de betalactamasas y mutaciones asociadas a la resistencia a quinolonas. Además, se observaron aislamientos de un mismo patotipo agrupados en distintos filogrupos. El análisis de genómica comparativa mostró un mayor número de genes ortólogos en aislamientos que pertenecían al mismo patotipo y filogrupo. Sobre la base de lo estudiado, los aislamientos de DEC en Lima, Perú, presentan resistencia a múltiples fármacos, y se detectaron varios patotipos y filogrupos con diversidad molecular y filogenética.

ABSTRACT In order to describe the genomic characteristics related to antimicrobial resistance and comparative genomics of diarrheagenic Escherichia coli (DEC), 14 DEC isolates from the strain collection of the Instituto Nacional de Salud (INS) were subjected to genome sequencing. We used bioinformatic procedures to analyze the obtained sequences in order to look for microbial resistance genes and genetic regions related to pathotypes and phylogroups. Several antimicrobial resistance determinants were detected, but the production of beta-lactamases and mutations associated to quinolone resistance were the most relevant. Additionally, we observed isolates of the same pathotype grouped in different phylogroups. The comparative genomics analysis showed a greater number of orthologous genes in isolates from the same pathotype and phylogroup. In conclusion, DEC isolates from Lima, Peru, showed resistance to multiple drugs; likewise, molecular and phylogenetic diversity was observed in several pathotypes and phylogroups.

Quinolones resistance in Salmonella spp. isolated from broilers and chickens carcasses under federal inspection / Resistência à quinolonas em Salmonella spp. isoladas de frangos vivos e carcaças sob inspeção federal

We analyzed 77 Salmonella spp. strains, from which 20 were isolated from broilers (cloacal swabs) and 57 from chickens from slaughterhouses under federal inspection. The following serotypes were identified: Salmonella Saint Paul (29), Salmonella Heidelberg (27), Salmonella Anatum (9), Salmonella Cerro (5), Salmonella Senftenberg (5), Salmonella enterica (O: 4,5) (1) and Salmonella enterica (O: 9.12) (1). Fifteen strains (19.5%) were resistant to enrofloxacin, six (7.8%) to ciprofloxacin, and 26 (33.8%) to nalidixic acid in the Disk Diffusion Test. The fifteen enrofloxacin resistant strains were selected for the PCR to detect the genes gyrA, gyrB, parC, and parE, and genetic sequencing to identify mutations in these genes. Five strains (33.3%) had point mutations in the gyrA gene, and one (6.7%) presented a point mutation in the parC gene. None of the 15 strains had mutations in the gyrB and parE genes, and none had more than one mutation in the gyrA gene or the other genes. The presence of point mutations in the strains studied corroborates with the phenotypic resistance observed to nalidixic acid. However, it did not explain the resistance to fluoroquinolones found in the 15 strains. Other mechanisms may be related to the fluoroquinolones resistance, highlighting the need for additional mutation screening.(AU)

Foram analisadas neste estudo 77 estirpes de Salmonella spp., 20 isoladas de frangos vivos (suabes de cloaca) e 57 isoladas de carcaças, provenientes de abatedouros frigoríficos sob Inspeção Federal. Foram identificados os seguintes sorotipos: Salmonella Saint Paul (29), Salmonella Heidelberg (27), Salmonella Anatum (9), Salmonella Cerro (5), Salmonella Senftenberg (5), Salmonella enterica (O: 4,5) (1) e Salmonella enterica (O: 9,12) (1). Do total de estirpes estudadas, 15 (19,5%) se mostraram resistentes à enrofloxacina, seis (7,8%) à ciprofloxacina e 26 (33,8%) ao ácido nalidíxico no Teste de Difusão em Disco. Foram selecionadas as 15 estirpes resistentes à enrofloxacina para a realização da PCR para detecção dos genes gyrA, gyrB, parC e parEe para sequenciamento genético do produto da PCR para identificação de mutações nesses genes. Cinco estirpes (33,3%) apresentaram mutações pontuais no gene gyrA e uma (6,7%) apresentou mutação pontual no gene parC. Nenhuma das 15 estirpes apresentou mutações nos genes gyrB e parE e nenhuma apresentou mais de uma mutação no gene gyrA ou nos outros genes. A existência apenas de mutações pontuais em alguns genes das estirpes analisadas está de acordo com a resistência fenotípica observada ao ácido nalidíxico, mas não explica a resistência às fluoroquinolonas encontrada nas 15 estirpes. Outros mecanismos de resistência podem estar relacionados à resistência encontrada às fluoroquinolonas e estudos adicionais são necessários para investigar sua presença.(AU)

Conduta terapêutica em pacientes com doença pulmonar obstrutiva crônica: monoterapia ou terapia cominada? / Therapy for chronic obstructive pulmonary disease patients: monotherapy or combination therapy?

Objetivo: Comparar a dupla terapia broncodilatadora com glicopirrônio mais indacaterol à monoterapia com glicopirrônio em pacientes portadores de doença pulmonar obstrutiva crônica. Métodos: Estudo clínico prospectivo, unicêntrico, controlado, cruzado, randomizado e duplo-cego realizado com 14 pacientes com diagnóstico de doença pulmonar obstrutiva crônica grau II. Os participantes receberam cada um dos tratamentos durante 30 dias. Antes de cada terapia, realizou-se período de wash-out por 7 dias, com broncodilador de curta ação. Antes e após cada intervenção, os pacientes passaram por exame de espirometria e responderam ao questionário COPD Assessment Test. Resultados: Observou-se melhora na função pulmonar medida por meio do volume expiratório forçado no primeiro segundo de 19mL (±36) para a monoterapia e 87mL (±33) para a terapia dupla. O ganho foi de 67mL (p=0,042) da associação dos medicamentos em relação ao glicopirrônio isolado. A melhora na qualidade de vida, medida a partir das pontuações do questionário, foi de 4,7 (±8,9) pontos para a monoterapia e 5,2 (±11) pontos para a dupla terapia (p=0,08). Conclusão: Ambos os tratamentos demonstram melhora na função pulmonar dos pacientes.

Objective: To compare dual bronchodilator therapy (Glycopyrronium with Indacaterol) versus Glycopyrronium monotherapy in patients with chronic obstructive pulmonary disease. Methods: This was a prospective, unicentric, controlled, crossover, randomized, and double-blind clinical trial with 14 patients diagnosed with grade II chronic obstructive pulmonary disease. The participants received each treatment during the period of 30 days. Before each therapy, a 7-day wash-out period with a short-acting bronchodilator was instituted. Before and after each intervention, the patients underwent spirometry and answered the COPD Assessment Test questionnaire. Results: An improvement in pulmonary function measured by forced expiratory volume during the first second of 19mL (±36) for monotherapy, and 87mL (±33) for dual therapy was observed. The gain was of 67mL (p=0.042) in the association of the drugs in relation to Glycopyrronium alone. The mean improvement in quality of life measured from the questionnaire scores was 4.7 (±8.9) points for monotherapy and 5.2 (± 11) points for dual therapy (p=0.08). Conclusion: Both treatments show improvement in the patients' pulmonary function.

Effect of focal adhesion kinase inhibition on osteoblastic cells grown on titanium with different topographies

Abstract Objective The present study aimed to investigate the participation of focal adhesion kinases (FAK) in interactions between osteoblastic cells and titanium (Ti) surfaces with three different topographies, namely, untreated (US), microstructured (MS), and nanostructured (NS). Methodology Osteoblasts harvested from the calvarial bones of 3-day-old rats were cultured on US, MS and NS discs in the presence of PF-573228 (FAK inhibitor) to evaluate osteoblastic differentiation. After 24 h, we evaluated osteoblast morphology and vinculin expression, and on day 10, the following parameters: gene expression of osteoblastic markers and integrin signaling components, FAK protein expression and alkaline phosphatase (ALP) activity. A smooth surface, porosities at the microscale level, and nanocavities were observed in US, MS, and NS, respectively. Results FAK inhibition decreased the number of filopodia in cells grown on US and MS compared with that in NS. FAK inhibition decreased the gene expression of Alp, bone sialoprotein, osteocalcin, and ALP activity in cells grown on all evaluated surfaces. FAK inhibition did not affect the gene expression of Fak, integrin alpha 1 ( Itga1 ) and integrin beta 1 ( Itgb1 ) in cells grown on MS, increased the gene expression of Fak in cells grown on NS, and increased the gene expression of Itga1 and Itgb1 in cells grown on US and NS. Moreover, FAK protein expression decreased in cells cultured on US but increased in cells cultured on MS and NS after FAK inhibition; no difference in the expression of vinculin was observed among cells grown on all surfaces. Conclusions Our data demonstrate the relevance of FAK in the interactions between osteoblastic cells and Ti surfaces regardless of surface topography. Nanotopography positively regulated FAK expression and integrin signaling pathway components during osteoblast differentiation. In this context, the development of Ti surfaces with the ability to upregulate FAK activity could positively impact the process of implant osseointegration.

Sistema microbiológico de cinco bioensayos para la detección de residuos de antibióticos / Five-assay microbiological system for the screeningf antibiotic residues

Abstract A novel microbiological system in microtiter plates consisting of five bioassays is presented for the detection and classification of antibiotic residues in milk. The bioassays were optimized for the detection of beta-lactams (Bioassay B: Geobacillus stearothermophilus), macrolides (Bioassay M: Bacillus megaterium with fusidic acid), tetracyclines (Bioassay T: B. megaterium with chloramphenicol), quinolones (Bioassay Q: Bacillus licheniformis) and sulfamides (Bioassay QS: B. licheniformis with trimethoprim) at levels near the maximum residue limits (MRL). The response of each bioassay was interpreted visually (positive or negative) after 4-5.5h of incubation. The system detects and classifies beta-lactams (5 pg/l of amoxicillin, 4 pg/l of ampicillin, 36 pg/l of cloxacillin, 22 pg/l of amoxicillin, 3 pg/l of penicillin, 114 pg/l of cephalexin, 89pg/l of cefoperazone and 116 pg/l of ceftiofur), tetracyclines (98 pg/l of chlortetracycline, 92 pg/l of oxytetracycline and 88 pg/l of tetracycline), macrolides (33 pg/l of erythromycin, 44 pg/l of tilmicosin and 50 pg/l of tylosin), sulfonamides (76 pg/l of sulfadiazine, 85 pg/l of sulfadimethoxine, 77 pg/l of sulfamethoxazole and 87pg/l of sulfathiazole) and quinolones (94 pg/l of ciprofloxacin, 98 pg/l of enrofloxacin and 79 pg/l marbofloxacin). In addition, the specificity values were high for B, T, Q (99.4%), M (98.8%) and QS (98.1%) bioassays. The control of antibiotics through this system can contribute to improving the quality and safety of dairy products.

Resumen Se presenta un novedoso sistema microbiológico en placas de microtitulación compuesto por 5 bioensayos para la detección y clasificación de residuos de antibióticos en leche. Los bioensayos fueron optimizados para la detección de betalactámicos (bioensayo B: Geobacillus stearothermophilus), macrólidos (bioensayo M: Bacillus megaterium con ácido fusídico), tetraciclinas (bioensayo T: Bacillus megaterium con cloranfenicol), quinolonas (bioensayo Q: Bacillus licheniformis) y sulfamidas (bioensayo QS: Bacillus licheniformis con trimetoprima), a niveles cercanos a los límites máximos de residuos (LMR). La respuesta de cada bioensayo se interpretó visualmente (positiva o negativa) después de 4 a 5,5 h de incubación. El sistema detecta y clasifica betalactámicos (5 pg/l de amoxicilina, 4 pg/l de ampicilina, 36 pg/l de cloxacilina, 22 pg/l de amoxicilina, 3 pg/l de penicilina, 114 pg/l de cefalexina, 89 pg/l de cefoperazona y 116 pg/l de ceftiofur), tetraciclinas (98 pg/l de clortetraciclina, 92 pg/l de oxitetraciclina y 88 pg/l de tetraciclina), macrólidos (33 pg/l de eritromicina, 44 pg/l de tilmi-cosina y 50 pg/l de tilosina), sulfamidas (76 pg/l de sulfadiacina, 85 pg/l de sulfadimetoxina, 77 pg/l de sulfametoxazol y 87 pg/l de sulfatiazol) y quinolonas (94 pg/l de ciprofloxacina, 98 pg/l de enrofloxacina y 79pg/l de marbofloxacina). Además, los valores de especificidad fueron altos para los bioensayos B, T, Q (99,4%), M (98,8%) y QS (98,1%). El control de residuos de antibióticos mediante este sistema puede contribuir a mejorar la calidad e inocuidad de los productos lácteos.

La leptina promueve la expresión de Hic-5 y la formación de puntos de actina por la vía dependiente de FAK-Src en células epiteliales mamarias MCF10A / Leptin induced Hic-5 expression and actin puncta formation by the FAK/Src-dependent pathway in MCF10A mammary epithelial cells

Resumen Introducción. La leptina es una hormona secretada por los adipocitos que se ha relacionado con el proceso de la transición de epitelio a mesénquima (Epithelial- Mesenchymal Transition, EMT). Promueve la migración e invasión de las células del epitelio mamario mediante la activación de las cinasas FAK y Src, un complejo regulador de vías de señalización que favorecen la expresión de las proteínas relacionadas con la formación de estructuras proteolíticas implicadas en la invasión y progresión del cáncer. Recientemente, se ha descrito que la sobreexpresión y activación de la proteína Hic-5 durante el mencionado proceso de transición, favorece la formación de los puntos de actina (indicativa de la formación y funcionalidad de los invadopodios), lo cual promueve la degradación local de los componentes de la matriz extracelular y la metástasis del cáncer. Objetivos. Evaluar el papel de las cinasas FAK y Src sobre la expresión y localización subcelular de Hic-5 y la formación de puntos de actina inducida por la leptina en la línea celular MCF10A de epitelio mamario no tumoral. Materiales y métodos. Se utilizaron los inhibidores específicos de la FAK (PF-573228) y la Src (PP2) para evaluar el papel de ambas cinasas en los niveles de expresión y localización subcelular de la proteína Hic-5 mediante Western blot e inmunofluorescencia, así como la formación de puntos de actina mediante la tinción con faloidina-TRITC en células MCF10A estimuladas con leptina. Resultados. La leptina indujo el incremento en la expresión de Hic-5 y la formación de puntos de actina. El tratamiento previo con los inhibidores de las cinasas FAK (PF-573228) y Src (PP2), promovió la disminución en la expresión de Hic-5 y de los puntos de actina en la línea celular MCF10A de epitelio mamario no tumoral. Conclusión. La leptina indujo la expresión y la localización perinuclear de Hic-5 y la formación de puntos de actina mediante un mecanismo dependiente de la actividad de las cinasas FAK y Src en las células MCF10A.

Abstract Introduction: Leptin is a hormone secreted by adipocytes that has been associated with the epithelial-mesenchymal transition (EMT). Additionally, leptin promotes the migration and invasion of mammary epithelial cells through the activation of FAK and Src kinases, which are part of a regulatory complex of signaling pathways that promotes the expression of proteins related to the formation of proteolytic structures involved in the invasion and progression of cancer. Recently, overexpression and activation of Hic-5 during the EMT have been shown to induce the formation of actin puncta; these structures are indicative of the formation and functionality of invadopodia, which promote the local degradation of extracellular matrix components and cancer metastasis. Objective: To evaluate the role of FAK and Src kinases in the expression of Hic-5 during the epithelial-mesenchymal transition induced by leptin in MCF10A cells. Materials and methods: We used specific inhibitors of FAK (PF-573228) and Src (PP2) to evaluate Hic-5 expression and subcellular localization by Western blot and immunofluorescence assays and to investigate the formation of actin puncta by epifluorescence in MCF10A cells stimulated with leptin. Results: Leptin induced an increase in Hic-5 expression and the formation of actin puncta. Pretreatment with inhibitors of FAK (PF-573228) and Src (PP2) promoted a decrease in Hic-5 expression and actin puncta formation in the non-tumorigenic mammary epithelial cell line MCF10A. Conclusion: In MCF10A cells, leptin-induced Hic-5 expression and perinuclear localization, as well as the formation of actin puncta through a mechanism dependent on the kinase activity of FAK and Src.

Antimicrobial susceptibility and minimal inhibitory concentration of bacteria isolated from the eyes of dogs with keratoconjunctivitis sicca / Susceptibilidade antimicrobiana e concentração inibitória mínima de bactérias isoladas dos olhos de cães com ceratoconjuntivite seca

The objective of this study was to evaluate the antimicrobial susceptibility profile of bacteria isolated from the eyes of dogs with keratoconjunctivitis sicca (KCS). We evaluated 65 dogs diagnosed with KCS and 30 healthy dogs (Control Group). Conjunctival swab samples were collected after KCS was diagnosed. Microbiological examinations were performed, including aerobic culture, antimicrobial susceptibility testing and minimum inhibitory concentration (MIC) determination for chloramphenicol, tobramycin, ofloxacin and moxifloxacin. MICs of the fifteen most resistant strains of Staphylococcus pseudintermedius (Staphylococcus intermedius Group, SIG) and the fifteen most resistant strains of gram-negative bacteria were determined. By percentage, the microorganisms exhibited the highest susceptibility to polymyxin B, tobramycin and chloramphenicol and the lowest to tetracycline. Three multi-drug-resistant strains of SIG were detected: one displayed isolated susceptibility to cefazolin, another to vancomycin, and another to polymyxin B and amikacin. The species of bacteria isolated from the eyes of dogs with KCS presented variable susceptibility to the antibiotics tested. We found evidence of the emergence of quinolone-resistant strains of SIG and further evidence of increased ocular prevalence. These findings reinforce the need to identify the bacteria involved and their antimicrobial susceptibility profile, as secondary infections can serve as exacerbating and perpetuating factors in KCS.(AU)

O objetivo deste estudo foi avaliar o perfil de susceptibilidade antimicrobiana de bactérias isoladas de olhos de cães com ceratoconjuntivite seca (CCS). Foram avaliados 65 cães com diagnóstico de CCS e 30 cães saudáveis ​​(Grupo Controle). Depois do diagnosticado de CCS, suabes conjuntivais foram coletados. Exames microbiológicos foram realizados, incluindo cultura aeróbia, teste de susceptibilidade antimicrobiana e determinação da concentração inibitória mínima (CIM) para cloranfenicol, tobramicina, ofloxacina e moxifloxacina. Para determinar a CIM, foram selecionadas as quinze cepas mais resistentes de Staphylococcus pseudintermedius (Staphylococcus intermedius Group-SIG) e as quinze cepas mais resistentes de bactérias gram-negativas. Os microrganismos apresentaram maior suscetibilidade percentual à polimixina B, tobramicina e cloranfenicol e menor suscetibilidade à tetraciclina. Três cepas de SIG resistentes a múltiplos medicamentos foram detectadas, do quais um demonstrou suscetibilidade isolada à cefazolina, outro à vancomicina e outro à polimixina B e à amicacina. As espécies de bactérias isoladas dos olhos de cães com CCS apresentaram suscetibilidade variável aos antibióticos testados. Encontramos evidências do surgimento de cepas resistentes à quinolona de S. pseudintermedius e outras evidências de aumento da prevalência ocular. Esses achados reforçam a necessidade de identificar as bactérias envolvidas e seu perfil de susceptibilidade aos antimicrobianos, pois as infecções secundárias podem servir como fatores exacerbantes e perpetuantes na CCS.(AU)

Marcadores de resistencia plasmídica a quinolonas qnr en aislamientos clínicos de enterobacterias productoras de betalactamasas CTX-M en Lima, Perú / Markers of plasmid resistance to qnr quinolones in clinical isolates of CTX-M beta-lactamases-producing enterobacteria in Lima, Peru

RESUMEN Con el objetivo de reportar marcadores de resistencia plasmídica a quinolonas qnr en aislamientos clínicos de enterobacterias productoras de betalactamasas CTX-M, se realizó un estudio descriptivo, con aislamientos del cepario del proyecto TO-06/09 del Instituto Nacional de Salud del Niño. Se recuperaron 138 aislamientos. La susceptibilidad antimicrobiana se determinó por el método de disco difusión y la identificación de genes por reacción en cadena de la polimerasa. De los 138 aislados, 67 (48,5%) fueron positivos para proteínas qnr por el método genotípico. De los cuales 38 (56,7%) presentaron determinantes qnrB y 48 (71,6%) determinantes qnrS. Ningún aislado presentó determinantes qnrA. Se detectó determinantes qnr en aislamientos que presentaban betalactamasas CTX-M en una población no expuesta.

ABSTRACT Aimed at reporting markers of plasmid resistance to qnr quinolones in clinical isolates of CTX-M beta-lactamase-producing enterobacteria, a descriptive study was conducted with isolates from the strain repository of TO-06/09 project of the National Children´s Health Institute. 138 isolates were recovered. Antimicrobial susceptibility was determined by the diffusion disk method, and gene identification by polymerase chain reaction. Of the 138 isolates, 67 (48.5%) were genotypically positive for qnr proteins; of these, 38 (56.7%) had qnrB determinants and 48 (71.6%) had qnrS determinants. No isolate presented qnrA determinants. qnr determinants were detected in isolates containing CTX-M beta-lactamases in a non-exposed population.

Ruptura espontánea del semitendinoso secundario al uso de una quinolona / Spontaneous semitendinosus rupture secondary to the use of a quinolone

Resumen La ruptura espontánea de un tendón secundario al uso de una quinolona es un efecto adverso poco común, pero que con el paso de los años se ha venido documentado con mayor frecuencia. A pesar de lo anterior, aún no hay estudios clínicos que permitan aclarar su fisiopatología, qué estrategias pueden disminuir el riesgo de desarrollar una ruptura espontánea o a qué dosis de las diferentes quinolonas se aumenta el riesgo de presentar una ruptura espontánea. Adicionalmente, varías guías de práctica clínica incentivan el uso de las quinolonas como primera línea para el manejo de infecciones respiratorias o de vías urinarias sin hacer consideraciones sobre este efecto adverso. Por lo anterior, presentamos a continuación el caso de un paciente de 31 años que posterior al inicio de ciprofloxacina para el manejo de una diarrea aguda presento una ruptura espontánea del tendón del semitendinoso secundario al uso de la quinolona. (Acta Med Colomb 2019; 44: 115-118).

Abstract The spontaneous rupture of a tendon secondary to the use of a quinolone is an uncommon adverse effect, but over the years has been documented more frequently. Despite this, there are still no clini cal studies to clarify its pathophysiology, nor which strategies can reduce the risk of developing a spontaneous rupture or at what dose of the different quinolones the risk of presenting a spontaneous rupture increases. In addition, several clinical practice guidelines encourage the use of quinolones as the first line for the management of respiratory or urinary tract infections without considering this adverse effect. Therefore, the case of a 31 year old patient who after the start of ciprofloxacin for the management of acute diarrhea had spontaneous semitendinosus tendon rupture secondary to the use of quinolone, is presented. (Acta Med Colomb 2019; 44: 115-118).

Susceptibilidad antimicrobiana en muestras urogenitales de ureaplasma urealyticum, el principal agente en infección intramaniótica / Antimicrobial susceptibility in urogenital samples of ureaplasma urealyticum, the main agent in intramaniotic infection

RESUMEN OBJETIVO: Ureaplasma urealyticum es el agente más frecuentemente aislado en infección intraamniótica. Los macrólidos son los antimicrobianos de primera elección en embarazadas. Se ha descrito el aumento de resistencia, pudiendo limitar las opciones terapéuticas durante la gestación. El propósito del estudio es evaluar susceptibilidad antimicrobiana de Ureaplasma urealyticum aislado en mujeres en edad fértil, que se atienden en Clínica Alemana Temuco, Araucanía, Chile. METODO: Se estudian todas las muestras de orina y flujo vaginal para cultivo de U. urealyticum, de pacientes entre 18 y 40 años, recibidas en el Laboratorio de Microbiología Clínica Alemana Temuco, en período Abril 2013 a Enero 2015. Se procesan las muestras con kit Mycoplasma IST 2 de Biomerieux. En las que resultan positivas, se estudia susceptibilidad a macrólidos, tetraciclinas y quinolonas. RESULTADOS: 426 muestras de orina y flujo vaginal (390 pacientes). 197 pacientes resultaron positivas para U. urealyticum. (50,5%). La susceptibilidad fue 88,4% (174 pctes) a Eritromicina, 87,9% (173 pctes) a Claritromicina y 91,9% (181 pctes) a Azitromicina (NS). 15 de 197 pacientes (7,6%) fueron resistentes a los 3 macrólidos. La susceptibilidad a Quinolonas fue 55,3% a Ciprofloxacino, y 94% a Ofloxacino. El 100% resultó susceptible a Tetraciclinas. CONCLUSIONES: Cerca del 10% de U. urealyticum aislados en nuestra serie son resistentes a macrólidos, contribuyendo a la no erradicación de la infección en tratamientos empíricos. Dentro de ellos, azitromicina aparece con la mayor efectividad. El aumento de resistencia limitará opciones terapéuticas, con gran impacto perinatal en futuro. La vigilancia de susceptibilidad en cada hospital es fundamental para elección terapéutica.

ABSTRACT INTRODUCTION: Ureaplasma urealyticum is the most frequently isolated microorganism in intra-amniotic infection. The macrolides are the first choice antimicrobials for treat this infection in pregnancy. The increasing resistance has been described worldwide, seriously limiting therapeutic options in pregnancy. The aim of the study is to evaluate antimicrobial susceptibility of U. urealyticum aislated in fertile-age women in Clínica Alemana Temuco, Araucania region, Chile. METHOD: Urine and vaginal samples were analyzed for U. urealyticum, from every 18 to 40 years old patients, received at Microbiology Laboratory of Clínica Alemana Temuco, between April 2013 to January 2015. The samples are processed with Mycoplasma IST 2 kit of Biomerieux. If they became positives, susceptibility to macrolides, tetracyclines and quinolones was studied. RESULTS: 426 urine and vaginal samples were collected (390 patients). 197 patients were positive for U. urealyticum (50.5%). The susceptibility was 88.4% (174 pts) to Erythromicyn, 87.9% (173 pts) to Clarithromycin and 91.9% (181 pts) to Azithromycin (NS). Resistance to all macrolides was observed in 15 out of 197 patients (7.6%). The susceptibility to Quinolones was 55.3% to Ciprofloxacin, and 94% to Ofloxacin. The 100% was susceptible to Tetracyclines. DISCUSSION: Near to 10% of isolated Ureaplasma spp in our serie were resistant to some macrolide, being a factor for failing to eradicate the infection in empirical treatment. Azithromycin was the most effective. The increasing resistance will limit therapeutic options, with great perinatal impact in the future. Susceptibility surveillance in each hospital is very important for therapeutic options.

Determination of antibiotic resistance genes and virulence factors in Escherichia coli isolated from Turkish patients with urinary tract infection

Abstract INTRODUCTION : Escherichia coli ranks among the most common sources of urinary tract infections (UTI). METHODS: Between November 2015 and August 2016, 90 isolates of E. coli were isolated from patients at Rize Education and Research Hospital in Turkey. Antibiotic susceptibility was determined for all isolates using the Kirby-Bauer disk diffusion method. These E. coli isolates were also screened for virulence genes, β-lactamase coding genes, quinolone resistance genes, and class 1 integrons by PCR. RESULTS: With respect to the antibiotic resistance profile, imipenem and meropenem were effective against 98% and 90% of isolates, respectively. A high percentage of the isolates showed resistance against β lactam/β lactamase inhibitor combinations, quinolones, and cephalosporins. PCR results revealed that 63% (57/90) of the strains carried class 1 integrons. In addition, a high predominance of extended-spectrum β-lactamases (ESBLs) was observed. The qnrA, qnrB, and qnrS genes were found in 24 (26.6%), 6 (6.6%), and 3 (3.3%), isolates, respectively. The most common virulence gene was fim (82.2%).The afa, hly, and cnf1 genes were detected in 16.6%, 16.6%, and 3.3% of isolates, respectively. Moreover, we observed eleven different virulence patterns in the 90 E. coli isolates. The most prevalent pattern was fım, while hly-fım, afa-aer-cnf-fım, aer-cnf, afa-aer, and afa-cnf-fım patterns were less common. CONCLUSIONS: Most of the E. coli virulence genes investigated in this study were observed in E. coli isolates from UTI patients. Virulence genes are very important for the establishment and maintenance of infection.

Clinical Aspects of Infectious Keratitis in Western Gyeongsangnamdo, Republic of Korea

PURPOSE: We investigated the regional characteristics and trends in causative agents, clinical features, and antibiotic susceptibility in infectious keratitis in western Gyeongnam province. METHODS: This retrospective chart review included 551 eyes of 551 patients with infectious keratitis, who were referred to our center from January 2004 to December 2017. The period of this study was divided into two terms of 7 years before and after 2011 to analyze the changes in causative organisms and antibiotic susceptibilities and to investigate the clinical features and regional characteristics in western Gyeongnam province. RESULTS: The most common occupation among patients was farming; the mean time taken for initial treatment was 8.6 days. The culture positivity rate was 35.8%, the most commonly isolated microorganisms were Staphylococcus epidermidis (14.5%) for Gram-positive bacteria and Pseudomona aeruginosa (13.5%) for Gram-negative bacteria. The distribution of culture-positive organisms before and after 2011 did not show any significant difference, but the increase in resistance to second and third generation quinolones was significantly greater in Gram-positive bacteria after 2011. There was no significant difference in clinical characteristics before and after 2011, but the hospital stay duration and treatment needs were significantly reduced. CONCLUSIONS: This was a large-scale study analyzing the clinical features of infectious keratitis in western Gyeognam province over a 14-year period. The results will help us understand the characteristics, microbiology, and community in infectious keratitis by analyzing patients referred to tertiary centers.

Antimicrobial Susceptibility and Genetic Analysis of Campylobacter jejuni Isolated from Diarrhea Patients in Busan / 대한임상미생물학회지

BACKGROUND: Campylobacter jejuni is an important food-borne pathogen that causes human gastroenteritis. This study was conducted to investigate the incidence of isolation, antimicrobial susceptibility pattern, and C. jejuni genotype from diarrhea patients in Busan, Korea. METHODS: A total of 97 C. jejuni were isolated from diarrhea patients during five food-borne outbreaks from 2014 to September 2017. Antimicrobial susceptibility tests were carried out by the broth microdilution method for ciprofloxacin (CIP), nalidixic acid (NAL), tetracycline (TET), chloramphenicol, azithromycin (AZI), erythromycin (ERY), streptomycin (STR), gentamicin, and telithromycin. To investigate C. jejuni genotypes, pulsed-field gel electrophoresis (PFGE) profile analysis was performed. RESULTS: The isolation rate of C. jejuni was 2.0% for the last 4 years and increased annually. Antimicrobial resistance rates of C. jejuni were shown to be in the order of NAL (90.9%), CIP (89.4%), TET (13.6%), AZI (3.0%), ERY (3.0%), and STR (1.5%). The proportion of multidrug-resistance was 18.2%, and they commonly contained quinolones (CIP-NAL). Analysis of PFGE patterns of SmaI-restricted DNA of C. jejuni isolates showed 17 clusters; cluster 11 was the major genotype pattern. CONCLUSION: This study will provide useful data for the proper use of antimicrobials and the management of resistant C. jejuni. Also it will help to provide data for the epidemiological investigation of foodborne diseases caused by C. jejuni, which is expected to increase in the future.

Pathologic characteristics for the Rhodococcus equi infection in foals in Jeju / 대한수의학회지

From 1996 to 2014, 14 foals from nine farms in Jeju were diagnosed with a Rhodococcus equi infection. Clinically, most foals showed characteristic respiratory signs, including hyperthermia and dyspnea. The seasonal occurrence of R. equi infection in foals was higher in summer, such as June (eight foals; 57.1%) and July (four foals; 28.6%), than in the other seasons. The major cases of R. equi infections were observed among two-month-old (eight foals; 57.1%) and three-month-old (three foals; 21.4%) foals. Histopathologically, bronchopneumonia, abscess, and granulomatous pneumonia were the most prevalent lesions in the lungs of foals. Colonic ulcers and submucosal abscesses were found in a foal. Some foals showed granulomatous lymphadenitis and abscesses in the mesenteric and other lymph nodes. According to the polymerase chain reaction using 10 tissue samples of foals and nine R. equi isolates, the vapA gene was detected in 11/11 (100%) foals. Immunohistochemical staining using the anti-VapA monoclonal antibody was applied to detect the R. equi VapA antigen in the organs of foals. R. equi VapA antigens were demonstrated in most lungs and some mesenteric and hilar lymph nodes of 13 foals. Isolated virulent R. equi VapA bacteria showed high sensitivity to gentamicin, quinolones, rifampin, and vancomycin.

Predictors of mortality in multidrug-resistant tuberculosis patients from Brazilian reference centers, 2005 to 2012

ABSTRACT Objectives To determine the main predictors of death in multidrug-resistant (MDRTB) patients from Brazil. Design Retrospective cohort study, a survival analysis of patients treated between 2005 and 2012. Results Of 3802 individuals included in study, 64.7% were men, mean age was 39 (1-93) years, and 70.3% had bilateral pulmonary disease. Prevalence of human immunodeficiency virus (HIV) was 8.3%. There were 479 (12.6%) deaths. Median survival time was 1452 days (4 years). Factors associated with increased risk of death were age greater than or equal to 60 years (hazard rate [HR] = 1.6, confidence interval [CI] = 1.15-2.2), HIV co-infection (HR = 1.46; CI = 1.05-1.96), XDR resistance pattern (HR = 1.74, CI = 1.05-2.9), beginning of treatment after failure (HR = 1.72, CI = 1.27-2.32), drug abuse (HR = 1.64, CI = 1.22-2.2), resistance to ethambutol (HR = 1.30, CI = 1.06-1.6) or streptomycin (HR = 1.24, CI = 1.01-1.51). Mainly protective factors were presence of only pulmonary disease (HR = 0.57, CI = 0.35-0.92), moxifloxacin use (HR = 0.44, CI = 0.25-0.80), and levofloxacin use (HR = 0.75; CI = 0.60-0.94). Conclusion A more comprehensive approach is needed to manage MDRTB, addressing early diagnostic, improving adhesion, and comorbidities, mainly HIV infection and drug abuse. The latest generation quinolones have an important effect in improving survival in MDRTB.