Bacteriological and molecular study of fosfomycin resistance in uropathogenic Escherichia coli.

The identification of genes associated with resistance has the potential to facilitate the development of novel diagnostic tests and treatment methods. The objective of this study was to examine the antibiotic resistance and Fosfomycin resistance genes in uropathogenic Escherichia coli (UPEC) in patients in Baghdad, Iraq. After analyzing 250 urine samples using various identification methods, including the examination of morphological characteristics, biochemical tests, and genetic detection, it was determined that E. coli was the most common bacteria present, accounting for 63.6% of the samples. Antibiotic susceptibility testing showed a significant prevalence of resistance to various antibiotics, with 99.3% of E. coli isolates exhibiting multiple drug resistance (MDR). Fosfomycin showed antibacterial properties against UPEC. The minimum inhibitory concentration (MIC) ranged from 512 to 1024 µg/mL, while the minimum bactericidal concentration (MBC) was 2048 µg/mL. In the time-kill assay, fosfomycin was effective against fosfomycin-resistant isolates within 8-12 h. The genetic determinants associated with fosfomycin resistance were examined through the utilization of polymerase chain reaction (PCR). The findings indicated that the genes murA, glpT, and cyaA were detected in all the isolates when genomic DNA was used as a template. However, all the tests yielded negative results when plasmid was used as a template. The genes fosA3 and fosA4 were detected in 8.6% and 5% of the isolates when genomic DNA was used as a template. When plasmid was used as a template, the genes fosA3 and fosA4 were found in 5.7% and 2.9% of the isolates, respectively. In conclusion, there is an increasing problem with antibiotic resistance in UPEC, with elevated rates of resistance to several antibiotics. The study also offers novel insights into the genetic foundation of fosfomycin resistance in UPEC.

Prevalence and antimicrobial resistance profile of pathogens isolated from patients with urine tract infections admitted to a university hospital in a medium-sized Brazilian city

ABSTRACT This study aimed to determine the antibiotic profile of microorganisms isolated from urine samples of patients with community urine tract infections (UTI) admitted to the University Hospital of the Federal University of Sao Carlos to support an appropriate local empirical treatment. A retrospective cross-sectional study was conducted from October 2018 to October 2020. Data from 1,528 positive urine cultures for bacterial pathogens and antibiograms were tabulated. Bacterial species prevalence and their resistance profile were analyzed and compared by sex and age. For Gram-negative fermenting bacteria, resistance rates were compared between patients with previous hospitalization and the total of infections caused by this group. For comparisons, the Chi-square test was performed, using Fisher's exact test when necessary (BioEstat program, adopting p ≤ 0.05). A multivariate analysis was applied to assess the effect of the studied variables in predicting multidrug resistance. Infections were more prevalent in women and older adults. Gram-negative bacteria represented 90.44% of total cultures. In both sexes, E. coli prevalence was significantly higher in adults compared with older adults (p < 0.0001). For several antibiotics, resistance rates were higher in the older adults compared with other ages and in patients with Gram-negative fermenting infections and previous hospitalization compared with the total of infections by this group of bacteria. The closer to the hospitalization, the higher the number of antibiotics with superior resistance rates. Resistance rates for aminoglycosides, carbapenems, ceftazidime, nitrofurantoin, piperacillin+tazobactam, and fosfomycin were less than 20%, considered adequate for empirical treatment. Only hospitalization in the previous 90 days was statistically significant in predicting infections by multidrug-resistant bacteria.

Prevalencia de bacterias multidrogoresistentes en un hospital público ubicado en la sierra del Perú

Introducción. La resistencia antibiótica es una de las mayores amenazas para la salud global. Una de las estrategias para su control, es la vigilancia microbiológica. Objetivo. Describir la variación de la prevalencia de cepas multidrogoresistentes (MDR) de las bacterias más frecuentemente aisladas en muestras clínicas de pacientes atendidos en un hospital de tercer nivel de una ciudad de altura en el Perú, y determinar los factores asociados a su aislamiento. Además, evaluar la prevalencia de otros fenotipos de resistencia. Métodos. Se realizó un estudio observacional transversal a partir de una cohorte histórica de aislamientos entre los años 2012 y 2019. Resultados. La prevalencia general de cepas MDR fue 74,1%, observándose una tendencia a la disminución de la prevalencia anual de cepas de MDR en cinco de las nueve bacterias analizadas. Los factores asociados a cepas MDR se correspondían con los descritos previamente: sexo masculino, edad mayor a 75 años y hospitalización en servicios de cuidados intensivos. Además, se observó un incremento en la prevalencia de otros fenotipos de resistencia. Conclusión. Se encontró una alta prevalencia de cepas MDR en todas las bacterias evaluadas, asociadas a factores previamente descritos.

Introduction. Antibiotic resistance is one of the greatest threats to global health. One of the strategies for its control is microbiological surveillance. Objective. To describe the variation of the prevalence of multidrug resistant strains (MDR) of the most frequently isolated bacteria in clinical samples of patients treated at a tertiary care hospital in a high-altitude city in Perú and the factors associated with its isolation. Also, to assess the prevalence of other resistance phenotypes. Results. The general prevalence of MDR strains was 74,1%, observing a downward trend in the annual prevalence of MDR strains in five of the nine bacteria included. The factors associated with MDR strains corresponded to those previously described: male sex, age over 75 years, and hospitalization in intensive care services. In addition, an increase in the annual prevalence of other resistance mechanisms was evidenced. Conclusions. A high prevalence of MDR strains was found in all the bacteria evaluated, associated with previously described factors.

Effect of Polyunsaturated Fatty Acids on Temozolomide Drug-Sensitive and Drug-Resistant Glioblastoma Cells.

Glioblastomas (GBMs) are notoriously difficult to treat, and the development of multiple drug resistance (MDR) is common during the course of the disease. The polyunsaturated fatty acids (PUFAs) gamma-linolenic acid (GLA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) have been reported to improve MDR in several tumors including breast, bladder, and leukaemia. However, the effects of PUFAs on GBM cell MDR are poorly understood. The present study investigated the effects of PUFAs on cellular responses to temozolomide (TMZ) in U87MG cells and the TMZ-resistant (TMZR) cells derived from U87MG. Cells were treated with PUFAs in the absence or presence of TMZ and dose-response, viable cell counting, gene expression, Western blotting, flow cytometry, gas chromatography-mass spectrometry (GCMS), and drug efflux studies were performed. The development of TMZ resistance caused an increase in ABC transporter ABCB1 and ABCC1 expression. GLA-, EPA-, and DHA-treated cells had altered fatty acid composition and accumulated lipid droplets in the cytoplasm. The most significant reduction in cell growth was seen for the U87MG and TMZR cells in the presence of EPA. GLA and EPA caused more significant effects on ABC transporter expression than DHA. GLA and EPA in combination with TMZ caused significant reductions in rhodamine 123 efflux from U87MG cells but not from TMZR cells. Overall, these findings support the notion that PUFAs can modulate ABC transporters in GBM cells.

Spectral analysis, in vitro cytotoxicity and antibacterial studies of bioactive principles from the leaves of Conocarpus lancifolius, a common tree of Jazan, Saudi Arabia / Análise espectral, citotoxicidade in vitro e estudos antibacterianos de princípios bioativos das folhas de Conocarpus lancifolius, uma árvore comum de Jazan, Arábia Saudita

The objective of the present study was to analyse the bioactive compounds of the leaves of Conocarpus lancifolius (C. lancifolius). The GC-MS analysis of the hot methanolic extract of the leaves (HMEL) of C. lancifolius exhibited the bioactive compounds such as 1-(3-Methoxy-2-nitrobenzyl) iso quinoline, morphin-4-ol-6,7-dione, 1-bromo N-methyl-, phytol, hexadecanoic acid, 2,3-dihydroxypropyl ester, 2,2:4,2”-terthiophene, ethyl iso-allocholate, caryophyllene oxide, campesterol, epiglobulol, cholestan-3-ol, 2-methylene-, (3á,5à)-, dasycarpidan-1-methanol, acetate (ester) and oleic acid, eicosyl ester. The FT-IR analysis of HMEL of C. lancifolius showed a unique peak at 3184, 2413, 1657 cm-¹ representing coumaric acid, chlorogenic acid and ferulic acid. The HMEL of C. lancifolius was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 72.66 ± 8.21 µg/ml as IC50 value. The HMEL of C. lancifolius also revealed a good spectrum of activity against Gram-positive and Gram negative bacterial cultures screened in this work. The activity observed has shown more or less similar effects against screened bacteria. However, the magnitude of potentiality was significantly lesser compared to standard ciprofloxacin disc at p< 0.001 level (99% confidence intervals). Furthermore, the study demonstrating the bioactive compounds can be isolated from the leaves of C. lancifolius.(AU)

O objetivo do presente estudo foi analisar os compostos bioativos das folhas de Conocarpus lancifolius (C. lancifolius). A análise por GC-MS do extrato metanólico quente das folhas (HMEL) de C. lancifolius exibiu os compostos bioativos como 1- (3-Metoxi-2-nitrobenzil) isoquinolina, morfina-4-ol-6,7- diona, 1-bromo-N-metil-, fitol, ácido hexadecanoico, 2,3-di-hidroxipropil éster, 2,2 ‘: 4, 2 ” - tertiofeno, isoalocolato de etil, óxido de cariofileno, campesterol, epiglobulol, colestano -3-ol, 2-metileno-, (3á, 5à) -, dasycarpidan-1-metanol, acetato (éster) e ácido oleico, éster eicosílico. A análise FT-IR de HMEL de C. lancifolius mostrou um pico único em 3184, 2413, 1657 cm-¹ representando ácido cumarico, ácido clorogênico e ácido ferúlico. O HMEL de C. lancifolius inibiu ativamente a proliferação de células de câncer de mama MCF-7 ATCC na concentração de 72,66 ± 8,21 µg/ml como valor de IC50. O HMEL de C. lancifolius também revelou bom espectro de atividade contra culturas de bactérias Gram-positivas e Gram-negativas rastreadas neste trabalho. A atividade observada mostrou efeitos mais ou menos semelhantes contra bactérias rastreadas. No entanto, a magnitude da potencialidade foi significativamente menor em comparação com o disco de ciprofloxacina padrão em nível de p < 0,001 (intervalos de confiança de 99%). Além disso, o estudo demonstrando os compostos bioativos pode ser isolado das folhas de C. lancifolius.(AU)

Spectral analysis, in vitro cytotoxicity and antibacterial studies of bioactive principles from the leaves of Conocarpus lancifolius, a common tree of Jazan, Saudi Arabia

Abstract The objective of the present study was to analyse the bioactive compounds of the leaves of Conocarpus lancifolius (C. lancifolius). The GC-MS analysis of the hot methanolic extract of the leaves (HMEL) of C. lancifolius exhibited the bioactive compounds such as 1-(3-Methoxy-2-nitrobenzyl) iso quinoline, morphin-4-ol-6,7-dione, 1-bromo-N-methyl-, phytol, hexadecanoic acid, 2,3-dihydroxypropyl ester, 2,2':4',2-terthiophene, ethyl iso-allocholate, caryophyllene oxide, campesterol, epiglobulol, cholestan-3-ol, 2-methylene-, (3á,5à)-, dasycarpidan-1-methanol, acetate (ester) and oleic acid, eicosyl ester. The FT-IR analysis of HMEL of C. lancifolius showed a unique peak at 3184, 2413, 1657 cm-1 representing coumaric acid, chlorogenic acid and ferulic acid. The HMEL of C. lancifolius was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 72.66 ± 8.21 µg/ml as IC50 value. The HMEL of C. lancifolius also revealed a good spectrum of activity against Gram-positive and Gram-negative bacterial cultures screened in this work. The activity observed has shown more or less similar effects against screened bacteria. However, the magnitude of potentiality was significantly lesser compared to standard ciprofloxacin disc at p 0.001 level (99% confidence intervals). Furthermore, the study demonstrating the bioactive compounds can be isolated from the leaves of C. lancifolius.

Resumo O objetivo do presente estudo foi analisar os compostos bioativos das folhas de Conocarpus lancifolius (C. lancifolius). A análise por GC-MS do extrato metanólico quente das folhas (HMEL) de C. lancifolius exibiu os compostos bioativos como 1- (3-Metoxi-2-nitrobenzil) isoquinolina, morfina-4-ol-6,7- diona, 1-bromo-N-metil-, fitol, ácido hexadecanoico, 2,3-di-hidroxipropil éster, 2,2 ': 4', 2 - tertiofeno, isoalocolato de etil, óxido de cariofileno, campesterol, epiglobulol, colestano -3-ol, 2-metileno-, (3á, 5à) -, dasycarpidan-1-metanol, acetato (éster) e ácido oleico, éster eicosílico. A análise FT-IR de HMEL de C. lancifolius mostrou um pico único em 3184, 2413, 1657 cm-1 representando ácido cumarico, ácido clorogênico e ácido ferúlico. O HMEL de C. lancifolius inibiu ativamente a proliferação de células de câncer de mama MCF-7 ATCC na concentração de 72,66 ± 8,21 µg / ml como valor de IC50. O HMEL de C. lancifolius também revelou bom espectro de atividade contra culturas de bactérias Gram-positivas e Gram-negativas rastreadas neste trabalho. A atividade observada mostrou efeitos mais ou menos semelhantes contra bactérias rastreadas. No entanto, a magnitude da potencialidade foi significativamente menor em comparação com o disco de ciprofloxacina padrão em nível de p 0,001 (intervalos de confiança de 99%). Além disso, o estudo demonstrando os compostos bioativos pode ser isolado das folhas de C. lancifolius.

Spectral analysis, in vitro cytotoxicity and antibacterial studies of bioactive principles from the leaves of Conocarpus lancifolius, a common tree of Jazan, Saudi Arabia / Análise espectral, citotoxicidade in vitro e estudos antibacterianos de princípios bioativos das folhas de Conocarpus lancifolius, uma árvore comum de Jazan, Arábia Saudita

Abstract The objective of the present study was to analyse the bioactive compounds of the leaves of Conocarpus lancifolius (C. lancifolius). The GC-MS analysis of the hot methanolic extract of the leaves (HMEL) of C. lancifolius exhibited the bioactive compounds such as 1-(3-Methoxy-2-nitrobenzyl) iso quinoline, morphin-4-ol-6,7-dione, 1-bromo-N-methyl-, phytol, hexadecanoic acid, 2,3-dihydroxypropyl ester, 2,2':4',2"-terthiophene, ethyl iso-allocholate, caryophyllene oxide, campesterol, epiglobulol, cholestan-3-ol, 2-methylene-, (3á,5à)-, dasycarpidan-1-methanol, acetate (ester) and oleic acid, eicosyl ester. The FT-IR analysis of HMEL of C. lancifolius showed a unique peak at 3184, 2413, 1657 cm-1 representing coumaric acid, chlorogenic acid and ferulic acid. The HMEL of C. lancifolius was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 72.66 ± 8.21 µg/ml as IC50 value. The HMEL of C. lancifolius also revealed a good spectrum of activity against Gram-positive and Gram-negative bacterial cultures screened in this work. The activity observed has shown more or less similar effects against screened bacteria. However, the magnitude of potentiality was significantly lesser compared to standard ciprofloxacin disc at p< 0.001 level (99% confidence intervals). Furthermore, the study demonstrating the bioactive compounds can be isolated from the leaves of C. lancifolius.

Resumo O objetivo do presente estudo foi analisar os compostos bioativos das folhas de Conocarpus lancifolius (C. lancifolius). A análise por GC-MS do extrato metanólico quente das folhas (HMEL) de C. lancifolius exibiu os compostos bioativos como 1- (3-Metoxi-2-nitrobenzil) isoquinolina, morfina-4-ol-6,7- diona, 1-bromo-N-metil-, fitol, ácido hexadecanoico, 2,3-di-hidroxipropil éster, 2,2 ': 4', 2 " - tertiofeno, isoalocolato de etil, óxido de cariofileno, campesterol, epiglobulol, colestano -3-ol, 2-metileno-, (3á, 5à) -, dasycarpidan-1-metanol, acetato (éster) e ácido oleico, éster eicosílico. A análise FT-IR de HMEL de C. lancifolius mostrou um pico único em 3184, 2413, 1657 cm-1 representando ácido cumarico, ácido clorogênico e ácido ferúlico. O HMEL de C. lancifolius inibiu ativamente a proliferação de células de câncer de mama MCF-7 ATCC na concentração de 72,66 ± 8,21 µg / ml como valor de IC50. O HMEL de C. lancifolius também revelou bom espectro de atividade contra culturas de bactérias Gram-positivas e Gram-negativas rastreadas neste trabalho. A atividade observada mostrou efeitos mais ou menos semelhantes contra bactérias rastreadas. No entanto, a magnitude da potencialidade foi significativamente menor em comparação com o disco de ciprofloxacina padrão em nível de p < 0,001 (intervalos de confiança de 99%). Além disso, o estudo demonstrando os compostos bioativos pode ser isolado das folhas de C. lancifolius.

Spectral analysis, in vitro cytotoxicity and antibacterial studies of bioactive principles from the leaves of Conocarpus lancifolius, a common tree of Jazan, Saudi Arabia / Análise espectral, citotoxicidade in vitro e estudos antibacterianos de princípios bioativos das folhas de Conocarpus lancifolius, uma árvore comum de Jazan, Arábia Saudita

The objective of the present study was to analyse the bioactive compounds of the leaves of Conocarpus lancifolius (C. lancifolius). The GC-MS analysis of the hot methanolic extract of the leaves (HMEL) of C. lancifolius exhibited the bioactive compounds such as 1-(3-Methoxy-2-nitrobenzyl) iso quinoline, morphin-4-ol-6,7-dione, 1-bromo N-methyl-, phytol, hexadecanoic acid, 2,3-dihydroxypropyl ester, 2,2’:4’,2”-terthiophene, ethyl iso-allocholate, caryophyllene oxide, campesterol, epiglobulol, cholestan-3-ol, 2-methylene-, (3á,5à)-, dasycarpidan-1-methanol, acetate (ester) and oleic acid, eicosyl ester. The FT-IR analysis of HMEL of C. lancifolius showed a unique peak at 3184, 2413, 1657 cm-¹ representing coumaric acid, chlorogenic acid and ferulic acid. The HMEL of C. lancifolius was actively inhibiting the proliferation of breast cancer cells MCF-7 ATCC at the concentration of 72.66 ± 8.21 µg/ml as IC50 value. The HMEL of C. lancifolius also revealed a good spectrum of activity against Gram-positive and Gram negative bacterial cultures screened in this work. The activity observed has shown more or less similar effects against screened bacteria. However, the magnitude of potentiality was significantly lesser compared to standard ciprofloxacin disc at p< 0.001 level (99% confidence intervals). Furthermore, the study demonstrating the bioactive compounds can be isolated from the leaves of C. lancifolius.

O objetivo do presente estudo foi analisar os compostos bioativos das folhas de Conocarpus lancifolius (C. lancifolius). A análise por GC-MS do extrato metanólico quente das folhas (HMEL) de C. lancifolius exibiu os compostos bioativos como 1- (3-Metoxi-2-nitrobenzil) isoquinolina, morfina-4-ol-6,7- diona, 1-bromo-N-metil-, fitol, ácido hexadecanoico, 2,3-di-hidroxipropil éster, 2,2 ‘: 4’, 2 ” - tertiofeno, isoalocolato de etil, óxido de cariofileno, campesterol, epiglobulol, colestano -3-ol, 2-metileno-, (3á, 5à) -, dasycarpidan-1-metanol, acetato (éster) e ácido oleico, éster eicosílico. A análise FT-IR de HMEL de C. lancifolius mostrou um pico único em 3184, 2413, 1657 cm-¹ representando ácido cumarico, ácido clorogênico e ácido ferúlico. O HMEL de C. lancifolius inibiu ativamente a proliferação de células de câncer de mama MCF-7 ATCC na concentração de 72,66 ± 8,21 µg/ml como valor de IC50. O HMEL de C. lancifolius também revelou bom espectro de atividade contra culturas de bactérias Gram-positivas e Gram-negativas rastreadas neste trabalho. A atividade observada mostrou efeitos mais ou menos semelhantes contra bactérias rastreadas. No entanto, a magnitude da potencialidade foi significativamente menor em comparação com o disco de ciprofloxacina padrão em nível de p < 0,001 (intervalos de confiança de 99%). Além disso, o estudo demonstrando os compostos bioativos pode ser isolado das folhas de C. lancifolius.

Eosinophilic cystitis, a rare cause of persistent haematuria in a diabetic dog / Cistite eosinofílica, uma causa rara de hematúria persistente em um cão diabético

Eosinophilic cystitis is a rare inflammatory disorder characterized by eosinophilic infiltration of entire layers of the bladder wall. The condition has been described in adults, children, and dogs. However, there are no consensus guidelines for the treatment of eosinophilic cystitis. Although human and veterinary literature reviews show some effectiveness in management with corticosteroids, antihistamines, and antibiotics, a variety of serious and frequent side effects are associated with steroid therapy. As a result, steroids are relatively contraindicated for patients with diabetes mellitus and Cushing's syndrome. A five-year-old neutered male chow-chow with controlled diabetes was referred with an 18-month history of malodorous urine, gross haematuria, and dysuria that were nonresponsive to antibiotics. The findings on general examination were unremarkable except for abdominal suprapubic discomfort. The complete blood count and biochemical profile (such as urea and creatinine) were normal except for mild peripheral eosinophilia. Although ultrasonography, bladder contrast radiography, and urine cytology findings indicated malignancy, with the presence of atypical urothelial cells, histopathology confirmed eosinophilic cystitis. Management with cyclosporine was adequate with complete remission of haematuria. This case report presents the first reported successful use of cyclosporine for the treatment of eosinophilic cystitis in a dog with diabetes.(AU)

A cistite eosinofílica é uma doença inflamatória rara caracterizada por infiltração eosinofílica de todas as camadas da parede da bexiga. Essa enfermidade já foi descrita em adultos, crianças e cães. No entanto, não há um consenso de diretrizes sobre o seu tratamento. Mesmo que as literaturas humana e veterinária mostrem alguma eficácia no manejo com corticosteroides, anti-histamínicos e antibióticos, uma variedade de efeitos colaterais graves e frequentes está associada à terapia com esteroides. Dessa forma, o uso de esteroides é relativamente contraindicado para pacientes com diabetes mellitus e síndrome de Cushing, por exemplo. Um chow-chow, macho, castrado, de cinco anos e diabético estável foi encaminhado para atendimento com histórico de urina fétida, hematúria macroscópica e disúria não responsiva a antibióticos há 18 meses. A avaliação dos parâmetros físicos estava dentro dos padrões, exceto por desconforto abdominal suprapúbico à palpação. O hemograma e o perfil bioquímico (como a ureia e a creatinina) estavam dentro da normalidade para a espécie, exceto por eosinofilia periférica leve. Embora a ultrassonografia, a radiografia contrastada da bexiga e os achados da urinálise indicassem malignidade, com a presença de células uroteliais atípicas, a histopatologia confirmou o diagnóstico definitivo de cistite eosinofílica. O manejo com ciclosporina foi satisfatório, com ausência completa da hematúria. Este relato de caso apresenta o primeiro uso documentado de ciclosporina para o tratamento de cistite eosinofílica com sucesso em um cão com diabetes.(AU)

Eosinophilic cystitis, a rare cause of persistent haematuria in a diabetic dog / Cistite eosinofílica, uma causa rara de hematúria persistente em um cão diabético

Eosinophilic cystitis is a rare inflammatory disorder characterized by eosinophilic infiltration of entire layers of the bladder wall. The condition has been described in adults, children, and dogs. However, there are no consensus guidelines for the treatment of eosinophilic cystitis. Although human and veterinary literature reviews show some effectiveness in management with corticosteroids, antihistamines, and antibiotics, a variety of serious and frequent side effects are associated with steroid therapy. As a result, steroids are relatively contraindicated for patients with diabetes mellitus and Cushing's syndrome. A five-year-old neutered male chow-chow with controlled diabetes was referred with an 18-month history of malodorous urine, gross haematuria, and dysuria that were nonresponsive to antibiotics. The findings on general examination were unremarkable except for abdominal suprapubic discomfort. The complete blood count and biochemical profile (such as urea and creatinine) were normal except for mild peripheral eosinophilia. Although ultrasonography, bladder contrast radiography, and urine cytology findings indicated malignancy, with the presence of atypical urothelial cells, histopathology confirmed eosinophilic cystitis. Management with cyclosporine was adequate with complete remission of haematuria. This case report presents the first reported successful use of cyclosporine for the treatment of eosinophilic cystitis in a dog with diabetes.(AU)

A cistite eosinofílica é uma doença inflamatória rara caracterizada por infiltração eosinofílica de todas as camadas da parede da bexiga. Essa enfermidade já foi descrita em adultos, crianças e cães. No entanto, não há um consenso de diretrizes sobre o seu tratamento. Mesmo que as literaturas humana e veterinária mostrem alguma eficácia no manejo com corticosteroides, anti-histamínicos e antibióticos, uma variedade de efeitos colaterais graves e frequentes está associada à terapia com esteroides. Dessa forma, o uso de esteroides é relativamente contraindicado para pacientes com diabetes mellitus e síndrome de Cushing, por exemplo. Um chow-chow, macho, castrado, de cinco anos e diabético estável foi encaminhado para atendimento com histórico de urina fétida, hematúria macroscópica e disúria não responsiva a antibióticos há 18 meses. A avaliação dos parâmetros físicos estava dentro dos padrões, exceto por desconforto abdominal suprapúbico à palpação. O hemograma e o perfil bioquímico (como a ureia e a creatinina) estavam dentro da normalidade para a espécie, exceto por eosinofilia periférica leve. Embora a ultrassonografia, a radiografia contrastada da bexiga e os achados da urinálise indicassem malignidade, com a presença de células uroteliais atípicas, a histopatologia confirmou o diagnóstico definitivo de cistite eosinofílica. O manejo com ciclosporina foi satisfatório, com ausência completa da hematúria. Este relato de caso apresenta o primeiro uso documentado de ciclosporina para o tratamento de cistite eosinofílica com sucesso em um cão com diabetes.(AU)

Small interfering RNA for cancer treatment: overcoming hurdles in delivery.

In many ways, cancer cells are different from healthy cells. A lot of tactical nano-based drug delivery systems are based on the difference between cancer and healthy cells. Currently, nanotechnology-based delivery systems are the most promising tool to deliver DNA-based products to cancer cells. This review aims to highlight the latest development in the lipids and polymeric nanocarrier for siRNA delivery to the cancer cells. It also provides the necessary information about siRNA development and its mechanism of action. Overall, this review gives us a clear picture of lipid and polymer-based drug delivery systems, which in the future could form the base to translate the basic siRNA biology into siRNA-based cancer therapies.

mcr-1-carrying Enterobacteriaceae isolated from companion animals in Brazil / Enterobacteriaceae portadora de mcr-1 em isolados clínicos de animais de companhia: primeiro relato no Brasil

Plasmid-mediated polymyxin resistance was first described in 2015, in China, in Escherichia coli carrying the mcr-1 (Mobile Colistin Resistance-1) gene. Since then, it has become a major public health challenge worldwide, representing a major threat to human and animal health. In addition, there are still few reports on the prevalence of mcr-1 in Enterobacteriaceae isolated from humans, animals and food. Therefore, the purpose of the study was to investigate the occurrence of the mcr-1 gene in bacterial isolates with phenotypic resistance to polymyxin B obtained from clinical specimens of companion animals. Phenotypic resistance to polymyxin B were determined by broth microdilution and the susceptibility profile to other antimicrobials (amikacin, amoxicillin/clavulanate, ampicillin, ampicillin/sulbactam, aztreonam, cefazolin, cefepime, cefotaxime, cefoxitin, ceftazidime, ceftriaxone, chloramphenicol, ciprofloxacin, doxycycline, ertapenem, gentamicin, imipenem, marbofloxacin, meropenem, phosphomycin, piperacillin/tazobactam, tetracycline, ticarcillin/clavulanate, tobramycin and trimethoprim/sulfamethoxazole) by disc-diffusion agar method. The extraction of bacterial DNA was performed via heat shock followed by spectrophotometric evaluation. To verify the presence of mcr-1, the Polymerase Chain Reaction was employed using specific primers, followed by agarose gel electrophoresis. The positive isolates had the corresponding amplicons sequenced. In this study, there were identified the first isolates of Escherichia coli, Klebsiella spp. and Enterobacter spp. carrying the mcr-1 gene derived from specimens of companion animals in Brazil. Our results suggest the dissemination of resistance to polymyxins in the community and the environment, highlighting the need for surveillance and optimized treatment guidelines.(AU)

A resistência à polimixina mediada por plasmídeo teve sua primeira descrição em 2015, na China, em Escherichia coli portadora do gene mcr-1 (Mobile Colistin Resistance-1) e a partir de então tornou-se um grande desafio para a saúde pública em todo o mundo, constituindo uma grande ameaça à saúde humana e animal. Além disso, ainda existem poucos relatos sobre a prevalência de mcr-1 em Enterobacteriaceae isoladas de humanos, animais e alimentos. Sendo assim, o objetivo do estudo foi investigar a ocorrência do gene mcr-1 em isolados bacterianos com resistência fenotípica à polimixina B, oriundos de materiais clínicos de animais de companhia. A resistência fenotípica à polimixina B foi determinada por microdiluição em caldo e o perfil de sensibilidade aos demais antimicrobianos (amicacina, amoxicilina/clavulanato, ampicilina, ampicilina/sulbactam, aztreonam, cefazolina, cefepime, cefotaxima, cefoxitina, ceftazidima, ceftriaxona, cloranfenicol, ciprofloxacina, doxiciclina, ertapenem, gentamicina, imipinem, marbofloxacino, meropenem, fosfomicina, piperacilina/tazobactam, tetraciclina, ticarcilina/clavulanato, tobramicina sulfametoxazol/trimetoprim) foram determinados pelo método disco difusão. A extração do DNA bacteriano foi realizada via choque térmico, seguido de avaliação espectrofotométrica. Para a verificação da presença do mcr-1 foi utilizada a Reação em Cadeia da Polimerase com emprego de iniciadores específicos, seguida de eletroforese em gel de agarose. Os isolados positivos tiveram os correspondentes amplicons sequenciados. Nesse estudo foram identificados os primeiros isolados de Escherichia coli, Klebsiella spp. e Enterobacter spp. portadores do gene mcr-1 derivados de espécimes de animais de companhia no Brasil. Este estudo sugere a disseminação da resistência às polimixinas na comunidade e no meio ambiente, destacando a necessidade de vigilância e diretrizes otimizadas de tratamento.(AU)

mcr-1-carrying Enterobacteriaceae isolated from companion animals in Brazil / Enterobacteriaceae portadora de mcr-1 em isolados clínicos de animais de companhia: primeiro relato no Brasil

Plasmid-mediated polymyxin resistance was first described in 2015, in China, in Escherichia coli carrying the mcr-1 (Mobile Colistin Resistance-1) gene. Since then, it has become a major public health challenge worldwide, representing a major threat to human and animal health. In addition, there are still few reports on the prevalence of mcr-1 in Enterobacteriaceae isolated from humans, animals and food. Therefore, the purpose of the study was to investigate the occurrence of the mcr-1 gene in bacterial isolates with phenotypic resistance to polymyxin B obtained from clinical specimens of companion animals. Phenotypic resistance to polymyxin B were determined by broth microdilution and the susceptibility profile to other antimicrobials (amikacin, amoxicillin/clavulanate, ampicillin, ampicillin/sulbactam, aztreonam, cefazolin, cefepime, cefotaxime, cefoxitin, ceftazidime, ceftriaxone, chloramphenicol, ciprofloxacin, doxycycline, ertapenem, gentamicin, imipenem, marbofloxacin, meropenem, phosphomycin, piperacillin/tazobactam, tetracycline, ticarcillin/clavulanate, tobramycin and trimethoprim/sulfamethoxazole) by disc-diffusion agar method. The extraction of bacterial DNA was performed via heat shock followed by spectrophotometric evaluation. To verify the presence of mcr-1, the Polymerase Chain Reaction was employed using specific primers, followed by agarose gel electrophoresis. The positive isolates had the corresponding amplicons sequenced. In this study, there were identified the first isolates of Escherichia coli, Klebsiella spp. and Enterobacter spp. carrying the mcr-1 gene derived from specimens of companion animals in Brazil. Our results suggest the dissemination of resistance to polymyxins in the community and the environment, highlighting the need for surveillance and optimized treatment guidelines.(AU)

A resistência à polimixina mediada por plasmídeo teve sua primeira descrição em 2015, na China, em Escherichia coli portadora do gene mcr-1 (Mobile Colistin Resistance-1) e a partir de então tornou-se um grande desafio para a saúde pública em todo o mundo, constituindo uma grande ameaça à saúde humana e animal. Além disso, ainda existem poucos relatos sobre a prevalência de mcr-1 em Enterobacteriaceae isoladas de humanos, animais e alimentos. Sendo assim, o objetivo do estudo foi investigar a ocorrência do gene mcr-1 em isolados bacterianos com resistência fenotípica à polimixina B, oriundos de materiais clínicos de animais de companhia. A resistência fenotípica à polimixina B foi determinada por microdiluição em caldo e o perfil de sensibilidade aos demais antimicrobianos (amicacina, amoxicilina/clavulanato, ampicilina, ampicilina/sulbactam, aztreonam, cefazolina, cefepime, cefotaxima, cefoxitina, ceftazidima, ceftriaxona, cloranfenicol, ciprofloxacina, doxiciclina, ertapenem, gentamicina, imipinem, marbofloxacino, meropenem, fosfomicina, piperacilina/tazobactam, tetraciclina, ticarcilina/clavulanato, tobramicina sulfametoxazol/trimetoprim) foram determinados pelo método disco difusão. A extração do DNA bacteriano foi realizada via choque térmico, seguido de avaliação espectrofotométrica. Para a verificação da presença do mcr-1 foi utilizada a Reação em Cadeia da Polimerase com emprego de iniciadores específicos, seguida de eletroforese em gel de agarose. Os isolados positivos tiveram os correspondentes amplicons sequenciados. Nesse estudo foram identificados os primeiros isolados de Escherichia coli, Klebsiella spp. e Enterobacter spp. portadores do gene mcr-1 derivados de espécimes de animais de companhia no Brasil. Este estudo sugere a disseminação da resistência às polimixinas na comunidade e no meio ambiente, destacando a necessidade de vigilância e diretrizes otimizadas de tratamento.(AU)

Antimicrobial Resistance in Nontyphoidal Salmonella Isolates from Human and Swine Sources in Brazil: A Systematic Review of the Past Three Decades.

Salmonella is the leading cause of foodborne illnesses worldwide. The widespread use of antimicrobials as prophylactic, therapeutic, and growth promoters in both livestock and human medicine has resulted in selective pressure regarding antimicrobial-resistant (AMR) bacteria. This systematic review summarizes phenotypic antimicrobial resistance profiles in Salmonella isolates from human and swine sources between 1990 and 2018 in Brazil. The 20 studies that matched the eligibility criteria-isolates from pigs and humans from Brazil, between 1990 and 2016, containing information on the number of Salmonella isolates, and applying the disk diffusion susceptibility method-were included. During the assessed period, Salmonella strains isolated from swine sources displayed the highest resistance rates for tetracycline (20.3%) and sulfonamides (17.4%). In contrast, human isolates displayed the highest resistance rates against ampicillin (19.8%) and tetracycline (17%). Salmonella Typhimurium was the most frequent AMR isolate from both swine and human sources, corresponding to 67% of all isolates. From 2001 to 2005, tetracycline and ampicillin were the top antimicrobial resistance compounds, and the most frequently detected in swine and human sources, respectively. A total of 63 and 58 multiple drug resistance profiles were identified in swine and human isolates, respectively. Antimicrobial resistance has decreased throughout the 1990-2016 period, except for gentamicin and nalidixic acid in swine and human isolates, respectively. The results indicate that Salmonella isolated from human and swine display resistance against clinically important antimicrobials, indicating that swine are possibly one of the main vectors for spreading human salmonellosis in Brazil.

Plasmid-mediated mcr-1 gene in acinetobacter baumannii and pseudomonas aeruginosa: first report from pakistan

Abstract INTRODUCTION: The increased use of colistin against infections caused by Acinetobacter baumannii and Pseudomonas aeruginosa has resulted in colistin resistance. The purpose of this study was to detect plasmid-mediated mcr-1 gene in colistin-resistant A. baumannii and P. aeruginosa isolates. METHODS: A total of 146 clinical isolates of A. baumannii (n = 62) and P. aeruginosa (n = 84) were collected from the four largest tertiary care hospitals in Peshawar, Pakistan. All bacterial isolates were phenotypically screened for multidrug resistance using the Kirby-Baur disc diffusion method. The minimum inhibitory concentration (MIC) of colistin in all isolates was phenotypically performed using dilution methods. mcr-1 gene was detected through polymerase chain reaction and the nucleotide sequence of amplicon was determined using Sanger sequencing. RESULTS: Approximately 96.7% A. baumannii and 83.3% P. aeruginosa isolates were resistant to multiple antibiotics. Colistin resistance was found in 9.6% (6/62) of A. baumannii and 11.9% (10/84) of P. aeruginosa isolates. Among 16 colistin resistant isolates, the mcr-1 gene was detected in one A. baumannii (1.61% of total isolates; 16.6% of colistin resistant isolates) and one P. aeruginosa strain (1.19% of total isolates; 10% of colistin resistant isolates). Nucleotide BLAST showed 98-99% sequence similarity to sequences of the mcr-1 gene in GenBank. CONCLUSIONS: Our study reports, for the first time, the emergence of plasmid-mediated mcr-1-encoded colistin resistance in multidrug resistant strains of A. baumannii and P. aeruginosa. Further large scales studies are recommended to investigate the prevalence of this mode of resistance in these highly pathogenic bacteria.

FK506 Attenuates the MRP1-Mediated Chemoresistant Phenotype in Glioblastoma Stem-Like Cells.

Poor response to current treatments for glioblastoma has been attributed to the presence of glioblastoma stem-like cells (GSCs). GSCs are able to expel antitumor drugs to the extracellular medium using the multidrug resistance-associated protein 1 (MRP1) transporter. Tacrolimus (FK506) has been identified as an MRP1 regulator in differentiated glioblastoma (GBM) cells (non-GSCs); however, the effect of FK506 on GSCs is currently unknown. The objective of the following research is to evaluate the effect of FK506 on the MRP1-related chemo-resistant phenotype of GSCs. For this, U87MG and C6 glioma cell lines were used to generate non-GSCs and GSCs. mRNA and MRP1-positive cells were evaluated by RT-qPCR and flow cytometry, respectively. A Carboxyfluorescein Diacetate (CFDA)-retention assay was performed to evaluate the MRP1 activity. Apoptosis and MTT assays were employed to evaluate the cytotoxic effects of FK506 plus Vincristine (MRP1 substrate). GSC-derived subcutaneous tumors were generated to evaluate the in vivo effect of FK506/Vincristine treatment. No differences in transcript levels and positive cells for MRP1 were observed in FK506-treated cells. Lesser cell viability, increased apoptosis, and CFDA-retention in the FK506/Vincristine-treated cells were observed. In vivo, the FK506/Vincristine treatment decreased the tumor size as well as ki67, Glial Fibrillary Acidic Protein (GFAP), and nestin expression. We conclude that FK506 confers a chemo-sensitive phenotype to MRP1-drug substrate in GSCs.

Comparative genome analysis and characterization of the Salmonella Typhimurium strain CCRJ_26 isolated from swine carcasses using whole-genome sequencing approach.

Salmonella pathogenicity relies on virulence factors many of which are clustered within the Salmonella pathogenicity islands. Salmonella also harbours mobile genetic elements such as virulence plasmids, prophage-like elements and antimicrobial resistance genes which can contribute to increase its pathogenicity. Here, we have genetically characterized a selected S. Typhimurium strain (CCRJ_26) from our previous study with Multiple Drugs Resistant profile and high-frequency PFGE clonal profile which apparently persists in the pork production centre of Rio de Janeiro State, Brazil. By whole-genome sequencing, we described the strain's genome virulent content and characterized the repertoire of bacterial plasmids, antibiotic resistance genes and prophage-like elements. Here, we have shown evidence that strain CCRJ_26 genome possible represent a virulence-associated phenotype which may be potentially virulent in human infection. SIGNIFICANCE AND IMPACT OF THE STUDY: Whole-genome sequencing technologies are still costly and remain underexplored for applied microbiology in Brazil. Hence, this genomic description of S. Typhimurium strain CCRJ_26 will provide help in future molecular epidemiological studies. The analysis described here reveals a quick and useful pipeline for bacterial virulence characterization using whole-genome sequencing approach.

Ventriculitis post-quirúrgica por Pseudomonas aeruginosa extensamente resistente tratada con colistina intratecal. Comunicación de un caso pediátrico y revisión de la literatura científica / Post-surgical ventriculitis due to extensively resistant Pseudomonas aeruginosa treated with intrathecal colistin. Pediatric case report and literature review

Resumen El uso de colistina por vía intratecal se ha consolidado como una opción terapéutica para el manejo de infecciones del sistema nervioso central causadas por bacilos gramnegativos multi-resistentes. La evidencia del éxito terapéutico y del perfil de seguridad es creciente, particularmente en infecciones por Acinetobacter baumanii multi-resistente en adultos. La evidencia en niños es escasa. Se presenta el caso clínico de una niña de 11 años de edad, con una ventriculitis post-quirúrgica por Pseudomonas aeruginosa extensamente resistente tratada con colistina intravenosa e intratecal. Se revisa su uso en niños con meningitis nosocomial causada por bacilos gramnegativos multi-resistentes.

Use of Intrathecal colistin has increased in recent years and has become an alternative for the management of infections of the central nervous system caused by multidrug resistant (MDR) bacteria. Evidence of therapeutic success and safety profile is increasing, particularly in MDR Acinetobacter baumanii infections in adults. Conversely, evidence in children is limited. We present a case of an 11-year-old female with postsurgical meningitis caused by an extensively resistant Pseudomonas aeruginosa strain and treated with venous and intrathecal colistin. The evidence of its use in children with nosocomial meningitis by MDR Gram negative bacteria is reviewed.

Characterization of multidrug-resistant Acinetobacter ssp. strains isolated from medical intensive care units in Cali - Colombia / Caracterización de cepas de Acinetobacter spp. resistentes a múltiples fármacos aisladas de unidades de cuidados intensivos en Cali - Colombia

Abstract Introduction: The extensive use of antibiotics has led to the emergence of multi-resistant strains in some species of the genus Acinetobacter. Objective: To investigate the molecular characteristics of multidrug-resistant of Acinetobacter ssp. strains isolated from 52 patients collected between March 2009 and July 2010 in medical intensive care units in Cali - Colombia. Methods: The susceptibility to various classes of antibiotics was determined by disc diffusion method, and the determination of the genomic species was carried out using amplified ribosomal DNA restriction analysis (ARDRA) and by sequencing of the 16s rDNA gene. Also, the genes of beta-lactamases as well as, integrases IntI1 and IntI2 were analyzed by PCR method. Results: The phenotypic identification showed that the isolates belong mainly to A. calcoaceticus- A. baumannii complex. All of them were multi-resistant to almost the whole antibiotics except to tigecycline and sulperazon, and they were grouped into five (I to V) different antibiotypes, being the antibiotype I the most common (50.0%). The percent of beta-lactamases detected was: blaTEM (17.3%), blaCTX-M (9.6%), blaVIM (21.2%), blaIMP (7.7%), blaOXA-58 (21.2%), and blaOXA-51 (21.2%). The phylogenetic tree analysis showed that the isolates were clustering to A. baumannii (74.1%), A. nosocomialis (11.1%) and A. calcoaceticus (7.4 %). Besides, the integron class 1 and class 2 were detected in 23.1% and 17.3% respectively. Conclusion: The isolates were identified to species A. baumanii mainly, and they were multiresistant. The resistance to beta-lactams may be by for presence of beta-lactamases in the majority of the isolates.

Resumen Introducción: El uso extensivo de antibióticos ha llevado a la emergencia de cepas multirresistentes en algunas especies del género Acinetobacter. Objetivo: Investigar las características moleculares de resistencia a múltiples fármacos de cepas aisladas de Acinetobacter spp. colectadas entre marzo de 2009 y julio de 2010 en 52 pacientes de unidades de cuidados intensivos en Cali - Colombia. Métodos: La susceptibilidad a diversas clases de antibióticos se determinó mediante el método de difusión de disco, y la determinación de la especie genómica se llevó a cabo usando un análisis de restricción de ADN ribosómico amplificado (ARDRA) y mediante la secuenciación del gen 16s de ADNr. Además, se analizaron por el método de PCR los genes de las beta-lactamasas, como también, las integrasas IntI1 e IntI2. Resultados: La identificación fenotípica mostró que los aislamientos pertenecen principalmente al complejo A. calcoaceticus - A. baumannii. Todos ellos eran multirresistentes a casi todos los antibióticos excepto tigeciclina y sulperazón, y se agruparon en cinco (I a V) antibitipos diferentes, siendo el antibiotipo I el más común (50%). El porcentaje de betalactamasas detectadas fue: blaTEM (17,3%), blaCTX-M (9,6%), blaVIM (21,2%), blaIMP (7,7%), blaOXA-58 (21,2%), blaOXA- 51 (21.2%). El análisis del árbol filogenético mostró que los aislados se agrupaban en A. baumannii (74.1%), A. nosocomialis (11.1%) y A. calcoaceticus (7.4%). Además, el integrón clase 1 y clase 2 se detectaron en 23.1% y 17.3% respectivamente. Conclusión: los aislamientos se identificaron principalmente como la especie A. baumanii, y fueron multirresistentes. La resistencia a los betalactámicos puede deberse a la presencia de betalactamasas en la mayoría de los aislamientos.

Incidencia de bacterias multi-resistentes en unidades de cuidados intensivos de hospitales chilenos / Incidence of multi-resistant bacteria in Intensive Care Units of Chilean hospitals

Resumen Introducción: La vigilancia de incidencia de bacterias multi-resistentes es un indicador que permite estimar mejor la magnitud de la resistencia bacteriana en los servicios hospitalarios. Objetivo: Evaluar la incidencia de bacterias multi-resistentes relevantes en unidades de cuidados intensivos del país y establecer las diferencias entre población adulta y pediátrica. Metodología: Se solicitó a los hospitales participantes información del número de aislados de siete bacterias multi-resistentes epidemiológicamente relevantes de unidades de cuidados intensivos (UCI) de adulto y pediátrico entre enero de 2014 y octubre de 2015, y el número de días-cama ocupados en dichas unidades en el mismo período. Con estos datos se calculó incidencia por 1.000 pacientes-día para cada unidad. Resultados: Se recibió información de 20 UCI adultos y 9 UCI pediátricas. En UCI adultos las bacterias de mayor incidencia fueron K. pneumoniae productora de BLEE [4,72 × 1.000 días cama (1,21-13,89)] y S. aureus resistente a oxacilina [3,85 (0,71-12,66)]. En pediatría la incidencia fue menor, destacando K. pneumoniae productora de BLEE [2,71 (0-7,11)] y P. aeruginosa resistente a carbapenémicos [1,61 (0,31-9,25)]. Conclusión: Se observan importantes diferencias entre los distintos hospitales en la incidencia de las bacterias estudiadas. La incidencia de bacterias multi-resistentes en UCI de adultos es significativamente mayor que en UCI pediátrica para la mayoría de las bacterias estudiadas.

Introduction: Incidence of multi-resistant bacteria is an indicator that permits better estimation of the magnitude of bacterial resistance in hospitals. Aim: To evaluate the incidence of relevant multi-drug resistant bacteria in intensive care units (ICUs) of Chile. Methods: Participating hospitals submitted information about the number of isolates from infected or colonized patients with 7 epidemiologically relevant multi-resistant bacteria in adult and pediatric ICUs between January 1, 2014 and October 31, 2015 and the number of bed days occupied in these units in the same period was requested. With these data incidence was calculated per 1,000 patient days for each unit. Results: Information from 20 adults and 9 pediatric ICUs was reviewed. In adult ICUs the bacteria with the highest incidence were K. pneumoniae ESBL [4.72 × 1,000 patient day (1.21-13.89)] and oxacillin -resistant S. aureus [3.85 (0.71-12.66)]. In the pediatric units the incidence was lower, highlighting K. pneumoniae ESBL [2.71 (0-7.11)] and carbapenem -resistant P. aeruginosa [1.61 (0.31-9.25)]. Conclusion: Important differences between hospitals in the incidence of these bacteria were observed. Incidence of multi-resistant bacteria in adult ICU was significantly higher than in pediatric ICU for most of the studied bacterias.