Abscesos renales bilaterales multifocales por Candida albicans / Bilateral multifocal kidney abscess due to Candida al bicans

Resumen Los abscesos renales son una complicación poco frecuente de las infecciones del tracto urinario y suelen asociarse con un aumento de la morbi-mortalidad. La mayoría de los casos ocurre en pacientes con factores predisponentes como la inmunosupresión. El diagnóstico requiere de una elevada sospecha clínica y el trata miento consiste en el uso de antibióticos y antifúngicos parenterales asociados o no a intervenciones quirúrgicas como nefrostomía y nefrectomía. Son pocos los casos publicados en la literatura médi ca de abscesos renales bilaterales multifocales y menos aún por Candida albicans. Se presenta el caso de una mujer de 20 años de edad con diabetes mellitus tipo 1 diagnosticada a los 8 años, múltiples internaciones por cetoacidosis diabética y reciente internación por can didemia (Candida albicans) completando tratamiento con fluconazol por 23 días. A los 18 días de su externación, consulta por dolor en flancos de tipo sordo y síntomas ge nerales; se realizó tomografía de abdomen con contraste que mostró abscesos multifocales bilaterales. Aislándose Candida albicans en una de las muestras obtenidas de las lesiones; recibió tratamiento con fluconazol 400 mg por 6 semanas endovenoso y 2 semanas vía enteral, evolu cionando favorablemente con mejoría clínica e image nológica continuando seguimiento clínico ambulatorio. Este reporte resalta la importancia del diagnóstico y tratamiento de esta complicación infrecuente en enfer medades complejas como la diabetes.

Abstract Renal abscesses are a rare complication of urinary tract infections and may be associated with increased morbidity and mortality. Most cases occur in patients with predisposing factors such as immunosuppression. Diagnosis requires high clinical suspicion and its treat ment consists in the use of parenteral antibiotics and antifungals associated or not with surgical interventions such as nephrostomy and nephrectomy. Few cases have been published in the medical literature of multifocal bilateral renal abscesses and even fewer due to Candida albicans. We present the case of a 20-year-old woman with type 1 diabetes mellitus, diagnosed at age 8, multiple hospitalizations for diabetic ketoacidosis, and recent hospitalization for candidemia (Candida albicans) treated with fluconazole for 23 days. Eighteen days after her discharge, she consulted for dull flank pain and gen eral symptoms. Contrast enhanced abdominal tomography showed bilateral multifocal abscesses and Candida albicans was isolated in one of the samples obtained from lesions. She received fluconazole 400 mg, 6 weeks i.v. and 2 weeks via enteral route, evolving favorably with clinical and imag ing improvement, continuing outpatient clinical monitoring. This report highlights the importance of diagnosis and treatment of this rare complication in complex diseases such as diabetes mellitus.

Validación de la metodología para cuantificar el fluconazol y sus impurezas orgánicas en materia prima por cromatografía líquida de alta resolución

Introducción. La eficiencia de una metodología para analizar una sustancia farmacológica puede verse afectada por las condiciones reales del laboratorio de cada país, incluyendo el clima. Por esta razón, se requiere validar el método con las pautas recomendadas para ello y optimizar el proceso, para asegurar el éxito y la confianza en los resultados. Objetivo. Validar una metodología para la cuantificación simultánea del fluconazol (materia prima) y sus impurezas orgánicas mediante cromatografía líquida de alta resolución con detector de arreglo de diodos en condiciones de clima tropical y con todos los requisitos normativos. Materiales y métodos. Se hicieron pruebas previas a la validación del método: idoneidad del sistema, estudio de filtros, límite de cuantificación, ausencia del error sistemático, estudios de degradación forzada y estabilidad de las soluciones. Además, se validaron: la especificidad, la linealidad, la exactitud, la precisión y la robustez. Resultados. La pureza espectral del método se logró al obtener la separación de los productos de degradación de los picos de los analitos. La estabilidad de las soluciones no se vio afectada, en la frecuencia evaluada de 24 horas, a temperatura ambiente y de refrigeración. Se obtuvo una linealidad con coeficientes de correlación mayores o iguales a 0,999 para la valoración y mayores o iguales a 0,997 para las impurezas. La recuperación estuvo en el rango de 98 a 102,0 % de fluconazol, con una exactitud entre el 80 y el 120 % para las impurezas. El factor de repetibilidad y reproducibilidad no superó la desviación estándar relativa del 2,0 % para la valoración y, la del 5,0 %, para las impurezas, lo cual mostró una solidez adecuada del método. Además, se obtuvo un tiempo corto de ejecución del análisis, lo que permitió la rápida determinación de la calidad de la materia prima. Conclusión. Se demostró que el método de cuantificación de fluconazol, validado por cromatografía líquida de alta resolución con detector de arreglo de diodos, es lo suficientemente selectivo, preciso, exacto, lineal y robusto; además, es capaz de generar resultados analíticos veraces en condiciones de uso reales, incluyendo el clima tropical de Colombia.

Introduction. The real laboratory conditions of each country, including climate, can affect the method's efficiency in analyzing a pharmacological substance. Thus, it is necessary to validate the process according to the corresponding guidelines and optimize it to ensure success and confidence in the results. Objective. The objective was to validate a methodology for fluconazole and its organic impurities quantification in raw material using high-performance liquid chromatography, with a diode array detector, under tropical climate conditions, and complying with all regulatory requirements. Materials and methods. We performed pre-validation tests of the method consisting of system adequacy, filters study, quantification limit, absence of systematic error, forced degradation studies, and solutions stability. In addition, we validated the specificity, linearity, accuracy, precision, and robustness of the system. Results. Separation of the degradation products from the analyte peaks allowed the achievement of the method's spectral purity. The solution's stability was not affected during the evaluated time (24 hours) at room temperature and under refrigeration. Linearity resulted in correlation coefficients greater than or equal to 0.999 for the evaluation and greater than or equal to 0.997 for impurities. We obtained a fluconazole recovery varying from 98 to 102% with an accuracy between 80 to 120% for impurities detection. The repeatability and reproducibility factor did not exceed a relative standard deviation of 2.0% for the evaluation and of 5.0% for the impurities, demonstrating the adequate robustness of the method. In addition, a short analysis execution time allowed the quick determination of the raw material quality. Conclusion. We demonstrated that the fluconazole quantification method validated by high-performance liquid chromatography is sufficiently selective, precise, exact, linear, and robust to generate accurate analytical results under real conditions, including the tropical climate of Colombia.

Expression of ERG11, ERG3, MDR1 and CDR1 genes in Candida tropicalis

Introduction. Drug resistance to azoles is a growing problem in the Candida genus. Objective. To analyze molecularly the genes responsible for fluconazole resistance in Candida tropicalis strains. Materials and methods. Nineteen strains, with and without exposure to fluconazole, were selected for this study. The expression of MDR1, CDR1, ERG11, and ERG3 genes was analyzed in sensitive, dose-dependent sensitive, and resistant strains exposed to different concentrations of the antifungal drug. Results. MDR1, ERG11 and ERG3 genes were significantly overexpressed in the different sensitivity groups. CDR1 gene expression was not statistically significant among the studied groups. Seven of the eight fluconazole-resistant strains showed overexpression of one or more of the analyzed genes. In some dose-dependent sensitive strains, we found overexpression of CDR1, ERG11, and ERG3. Conclusion. The frequency of overexpression of ERG11 and ERG3 genes indicates that they are related to resistance. However, the finding of dose-dependent resistant/sensitive strains without overexpression of these genes suggests that they are not exclusive to this phenomenon. More basic research is needed to study other potentially involved genes in the resistance mechanism to fluconazole.

Introducción. La farmacorresistencia a los azoles es un problema creciente en el género Candida. Objetivo. Analizar molecularmente los genes responsables de la resistencia a fluconazol en cepas de Candida tropicalis. Materiales y métodos. Para este estudio, se seleccionaron 19 cepas, con exposición a fluconazol y sin ella. Se analizó la expresión de los genes MDR1, CDR1, ERG11 y ERG3 en cepas sensibles, sensibles dependiente de la dosis, y resistentes, previamente expuestas a diferentes concentraciones del fármaco antifúngico. Resultados. Se encontró que los genes MDR1, ERG11 y ERG3 estaban significativamente sobreexpresados en los diferentes grupos de sensibilidad. La expresión del gen CDR1 no fue estadísticamente significativa entre los grupos estudiados. Siete de las ocho cepas resistentes a fluconazol mostraron sobreexpresión de uno o más de los genes analizados. En algunas cepas sensibles dependientes de la dosis, se encontró sobreexpresión de CDR1, ERG11 y ERG3. Conclusión. La sobreexpresión de los genes ERG11 y ERG3 indica que están relacionados con la resistencia de las cepas de Candida. Sin embargo, el hallazgo de cepas resistentes o sensibles según la dosis, sin sobreexpresión de estos genes, sugiere que pueden existir otros genes involucrados en este fenómeno. Se necesitan más investigaciones básicas que contribuyan al estudio de otros genes potencialmente involucrados en el mecanismo de resistencia al fluconazol.

A Rare Case Report of “Torsades De Pointes” Induced by Fluconazole-Levetiracetam Combined Therapy

Drugs used to treat cardiovascular disease as well as those used in the treatment of multiple other conditions can occasionally produce exaggerated prolongation of the QT interval on the surface electrocardiogram and the morphologically distinctive polymorphic ventricular tachycardia that results is known as «torsade de pointe». «Torsade de pointe» (TDP) is a characteristic polymorphic ventricular arrhythmia associated with delayed ventricular repolarization as evidenced on the surface electrocardiogram by QT interval prolongation. It typically occurs in self-limiting bursts, causing dizziness and syncope, but may occasionally progress to ventricular fibrillation and sudden death. This rare case report showed the potential higher risk of the occurrences of «Tdp» when levetiracetam (KEPPRA) was used in combination therapy with fluconazole, which is already a known medication with the risk of causing polymorphic ventricular arrhythmia.

Chemical constituents from Paris rugosa rhizomes and their antimicrobial activities / 中国中药杂志

Paris rugosa(Melanthiaceae) only grows in Yunnan province of China at present, and its chemical constituents have not been systematically studied. In this study, nine compounds, including one new compound pariposide G(1) and eight known compounds of cerin(2), stigmast-4-en-3-one(3), β-ecdysone(4), ophiopogonin C'(5), methyl protogracillin(6), gracillin(7), parissaponin H(8), and parisyunnanoside G(9), were isolated and identified from the ethanol extract of P. rugosa rhizomes by column chromatography methods and semi-preparative high-performance liquid chromatography(HPLC). Compounds 1-9 were isolated from this plant for the first time. The antibacterial and antifungal activities of all the compounds were evaluated. The results showed that ophiopogonin C' had strong inhibitory effects on Candida albicans [MIC_(90)=(4.68±0.01) μmol·L~(-1)] and the fluconazole-resistant strain of C. albicans [MIC_(90)=(4.66±0.02) μmol·L~(-1)].

Perfiles de expresión de los genes ERG11, MDR1 y AFR1 en Cryptococcus neoformans var. grubii aislados de pacientes con HIV / Gene expression profiles of ERG11, MDR1 y AFR1 in Cryptococcus neoformans var. grubbi isolated from HIV patients

Introducción. El fluconazol es el antifúngico más utilizado para la prevención y el tratamiento de infecciones causadas por el género Cryptococcus, agente etiológico de la criptococosis. La resistencia al fluconazol en los aislamientos de Cryptoccocus neoformans puede hacer fracasar el tratamiento y generar recaídas de la infección. Objetivo. Evaluar los perfiles de expresión de los genes AFR1, MDR1 y ERG11 en aislamientos clínicos de C. neoformans var. grubii, durante la respuesta in vitro a la inducción con fluconazol. Materiales y métodos. Se estudiaron 14 aislamientos de C. neoformans var. grubii provenientes de pacientes con HIV, de los cuales 6 eran sensibles al fluconaol y 8 presentaban sensibilidad disminuida. Los niveles de expresión de los genes ERG11, AFR1 y MDR1 se determinaron mediante PCR en tiempo real. Resultados. Los aislamientos resistentes al fluconazol mostraron sobreexpresión de los genes AFR1 y MDR1, mientras que la expresión de los fenotipos de resistencia evaluados se mantuvo homogénea en ERG11, en todos los aislamientos de C. neoformans var. grubii. Conclusiones. La sobreexpresión de los genes AFR1 y MDR1 que codifican las bombas de eflujo, contribuye a la resistencia al fluconazol en los aislamientos estudiados. Sin embargo, los patrones de resistencia que se registran en este hongo, sumado a los casos de recaídas en pacientes con HIV, no pueden atribuirse únicamente a los casos de resistencia por exposición al fármaco. Otros mecanismos podrían también estar involucrados en este fenómeno, como la resistencia emergente (resistencia mediante otros genes ERG) y la heterorresistencia, los cuales deben ser estudiados en estos aislamientos.

Introduction: Fluconazole is the most used antifungal drug for prevention and treatment of Cryptococcus spp. infections, the etiological agent of cryptococcosis. Resistance to fluconazole among Cryptococcus neoformans isolates can lead to treatment failure and generate relapses. Objective: To evaluate the expression profles of the AFR1, MDR1 and ERG11 genes in C. neoformans var. grubii clinical isolates during the in vitro response to fluconazole induction. Materials and methods: Fourteen C. neoformans var. grubii isolates recovered from HIV patients were studied, in which 6 showed sensitivities to fluconazole and 8 decreased sensitivity. The expression levels of ERG11, AFR1 and MDR1 genes were determined by real-time PCR from extracted mRNA. Results: AFR1 and MDR1 genes from C. neoformans var. grubii were overexpressed in fluconazole resistant isolates, whereas ERG11 maintains homogeneous expression in all the evaluated resistance phenotypes of C. neoformans var. grubii isolates. Conclusions: The overexpression of AFR1 and MDR1 genes, which codify for efflux pumps, contributes to fluconazole resistance in the studied isolates. However, the resistance patterns in this fungus and the relapse cases in HIV patients cannot be attributed solely to the exposure to the drug. Heteroresistance and the emerging resistance (resistance through other ERG genes), might be other mechanisms involved in this phenomenon, which must be studied in these isolations.

Evaluation of antifungal susceptibility pattern of dermatophytes isolated in a tertiary care hospital

Background: Superficial fungal infections are extremely common in the tropical and subtropical regions all over the world. Although numerous antifungal drugs are available for the treatment, the pattern of susceptibility to the drugs being used for dermatophytes varies with time and place. Aims and Objectives: The objective of the study was to isolate, identify, and determine antifungal susceptibility pattern in dermatophytes isolated from patients attending dermatology OPD in a tertiary care hospital. Materials and Methods: This study was conducted on 150 dermatophytes isolated from clinically diagnosed patients with dermatomycosis of skin and nail attending the Department of Dermatology Outpatient Clinic of JJM Medical College over a period of 1 year. These cases were referred to the Department of Microbiology for fungal isolation, culture and sensitivity testing. Results: The most frequently affected age group was 21–30 years. The most common clinical manifestation was tinea corporis (52.56%) followed by tinea cruris (26%). The most common dermatophytes implicated were Trichophyton species in 88%. Irrespective of the species, about 96% of our isolates exhibited high minimum inhibitory concentration (MIC) (>1 ?g/ml) to fluconazole. The MIC of luliconazole was < 0.004 ?g/ml to all the dermatophytes isolates tested, compared to other antifungal agents that were ?0.25 ?g/ml. The MIC range was narrowest for luliconazole 0.002–0.128 ?g/ml and the widest for fluconazole, itraconazole, and ketoconazole (0.125–64, 0.0321–16, and 0.0321–16, respectively). Conclusion: Our study showed that luliconazole was the most active drug against all dermatophytes isolates, followed by itraconazole, ketoconazole, and fluconazole. The higher and wider range in MIC values for itraconazole, ketoconazole, and fluconazole found for some of our isolates raise the possibility of increasing resistance to these drugs.

An epidemiologic analysis of Candida spp. urinary infections in intensive care unit / Uma análise epidemiológica das infecções urinárias por Candida spp. em unidade de terapia intensiva / Un análisis epidemiológico de las infecciones urinarias por Candida spp. en la unidad de cuidados intensivos

Background and objectives: The finding of Candida species in urine is an usual finding and is called candiduria. There is an increase in the frequency of urinary tract infections (UTI) caused by Candida especially in critically ill patients. This study aimed to determine the epidemiological, clinical, and mycological characteristics of Candida urinary infections in intensive care unit (ICU) and antifungal susceptibilities. Methods: Urine cultures of 394 ICU patients with clinical suspicion of UTI were evaluated. After 24-48 hours of incubation, colonies appeared to grow as yeast, were morphologically examined by Gram staining. Candida strains that grew 104 ≥ CFU/mL in urine cultures were accepted as candiduria. The susceptibilities of the Candida strains to amphotericin B, itraconazole, fluconazole, voriconazole, flucytosine, and caspofungin were investigated with broth microdilution method. Results: The distribution of the isolated 100 urinary Candida strains were as, 54 Candida albicans, 34 C. glabrata, 7 C. tropicalis, 2 C. kefyr, 2 C. lusitaniae, and 1 as C. parapsilosis. Among 100 Candida species isolated in our study susceptibility rates of amphotericin B, flucytosine, caspofungin, fluconazole, itraconazole, and voriconazole were 100%, 100%, 91%, 23%, 13%, 25.8%, respectively. Conclusion: Accurate identification of Candida spp., as well as the investigating the antifungal susceptibility, will be beneficial in terms of the effectiveness of the treatment and the prevention of resistance development.(AU)

Justificativa e objetivos: O achado de espécies de Candida na urina é um achado comum e é chamado de candidúria. Há um aumento na frequência de infecções do trato urinário (ITU) causadas por Candida, principalmente em pacientes críticos. Este estudo teve como objetivo determinar as características epidemiológicas, clínicas e micológicas das infecções urinárias por Candida em unidade de terapia intensiva (UTI) e a susceptibilidade aos antifúngicos. Métodos: Foram avaliadas culturas de urina de 394 pacientes de UTI com suspeita clínica de ITU. Após 24-48 horas de incubação, as colônias pareceram crescer como leveduras, foram morfologicamente examinadas por coloração de Gram. As cepas de Candida que cresceram ≥104 UFC/mL em culturas de urina foram aceitas como candidúria. As suscetibilidades das cepas de Candida à anfotericina B, itraconazol, fluconazol, voriconazol, flucitosina e caspofungina foram investigadas com o método de microdiluição em caldo. Resultados: A distribuição das cepas 100 isoladas de Candida urinária foi de 54 Candida albicans, 34 C. glabrata, 7 C. tropicalis, 2 C. kefyr, 2 C. lusitaniae e 1 como C. parapsilosis. Entre 100 espécies de Candida isoladas em nosso estudo, as taxas de susceptibilidade de anfotericina B, flucitosina, caspofungina, fluconazol, itraconazol e voriconazol foram de 100%, 100%, 91%, 23%, 13%, 25,8%, respectivamente. Conclusão: A identificação precisa de Candida spp., bem como a investigação da susceptibilidade aos antifúngicos, será benéfica em termos de eficácia do tratamento e prevenção do desenvolvimento de resistência.(AU)

Justificación y objetivos: El hallazgo de especies de Candida en la orina es un hallazgo habitual y se denomina candiduria. Hay un aumento en la frecuencia de infecciones del tracto urinario (ITU) causadas por Candida, especialmente en pacientes críticamente enfermos. Este estudio tuvo como objetivo determinar las características epidemiológicas, clínicas y micológicas de las infecciones urinarias por Candida en la unidad de cuidados intensivos (UCI) y la susceptibilidad antifúngica. Métodos: Se evaluaron urocultivos de 394 pacientes de UCI con sospecha clínica de ITU. Después de 24-48 horas de incubación, las colonias parecían crecer como levadura, se examinaron morfológicamente mediante tinción de Gram. Las cepas de Candida que crecieron 104 ≥ UFC / ml en urocultivos se aceptaron como candiduria. Las susceptibilidades de las cepas de Candida a la anfotericina B, itraconazol, fluconazol, voriconazol, flucitosina y caspofungina se investigaron con el método de microdilución en caldo. Resultados: La distribución de las cepas 100 urinarias aisladas de Candida fue de, 54 C. albicans, 34 C. glabrata, 7 C. tropicalis, 2 C. kefyr, 2 C. lusitaniae y 1 como C. parapsilosis. Entre las 100 especies de Candida aisladas en nuestro estudio, las tasas de susceptibilidad de anfotericina B, flucitosina, caspofungina, fluconazol, itraconazol y voriconazol fueron 100%, 100%, 91%, 23%, 13%, 25,8%, respectivamente. Conclusión: La identificación precisa de Candida spp., así como la investigación de la susceptibilidad antifúngica, será beneficiosa en términos de la eficacia del tratamiento y la prevención del desarrollo de resistencias.(AU)

Perfil de resistencia antifúngica en el tratamiento de candidiasis vaginal: Un diagnóstico de agentes etiológicos / Antifungal resistance profile in the treatment of vaginal candidiasis: A diagnosis of etiologic agents

Introducción: En las micosis, la resistencia a los antifúngicos aumenta debido a los diagnósticos y tratamientos incorrectos. Por tanto, se ha sugerido la vigilancia y el seguimiento de las cepas resistentes para garantizar una terapia adecuada. Objetivo: Determinar las especies de Candida y el perfil de resistencia a fluconazol y voriconazol, que presentan los aislados obtenidos de muestras almacenadas durante los meses diciembre 2017 y marzo 2018 de pacientes provenientes del Laboratorio del Hospital Regional "Miguel Ángel Mariscal Llerena" del departamento de Ayacucho, Perú. Material y Métodos: Estudio descriptivo transversal, en el cual se aislaron 110 cepas de Candida sp almacenadas por un período de cuatro meses, y procesadas por métodos estandarizados y aprobados por el Ministerio de Salud (MINSA) y el Instituto Nacional de Salud (INS) del Perú relacionados con el diagnóstico de agentes etiológicos de micosis humanas y la sensibilidad antifúngica con los métodos estandarizados de Clinical Laboratory Standard Institute (CLSI). Resultados: Se observó C. albicans en 86,4 % de los aislados seguida por C. glabrata con 9,1 %, C. parapsilosis 2,7 % y 0,9 % de C. tropicalis y C. krusei. El 10,5 % de C. albicans fue resistente al fluconazol y voriconazol con CMI ≥ 128 μg/mL y ≥ 16 μg/mL respectivamente, mientras que 20 % de C. glabrata mostró sensibilidad dosis dependiente y 10 % de resistencia al fluconazol. Conclusiones: Existe una gran variedad de especies de Candida, siendo la C. albicans la más común, seguida por C. glabrata, con un mayor porcentaje de aislamiento en comparación con otras especies. Se puede observar que estas especies poseen grados de vulnerabilidad considerables a la aplicación de fármacos como el fluconazol y voriconazo(AU)

Introduction: In mycoses, resistance to antifungals increases due to incorrect diagnosis and treatment. Therefore, surveillance and monitoring of resistant strains has been suggested to ensure adequate therapy. Objective: To determine the Candida species and the resistance profile to fluconazole and voriconazole presented by the isolates obtained from samples stored during the months of December 2017 and March 2018 from patients coming from the Laboratory of the Regional Hospital "Miguel Ángel Mariscal Llerena" in the department of Ayacucho, Peru. Material and Methods: Cross-sectional descriptive study in which 110 strains of Candida sp were isolated and stored for a period of four (04) months, and processed by standardized methods approved by the Ministry of Health (MINSA) and the National Institute of Health (INS) of Peru related to the diagnosis of etiological agents of human mycosis and antifungal sensitivity with the standardized methods of the Clinical Laboratory Standard Institute (CLSI). Results: C. albicans was observed in 86,4 % of isolates, followed by C. glabrata (9,1 %), C. parapsilosis (2,7 %), and 0,9 % of C. tropicalis and C. krusei; 10,5 % of C. albicans was resistant to fluconazole and voriconazole with MIC ≥ 128 μg/mL and ≥ 16 μg/mL respectively, while 20 % of C. glabrata showed dose dependent sensitivity and 10 % resistance to fluconazole. Conclusions: There is a wide variety of Candida species, with C. albicans being the most common, followed by C. glabrata, with a higher percentage of isolation compared to other species. It can be seen that these species have considerable degrees of vulnerability to the application of drugs such as fluconazole and voriconazole(AU)

Evaluation on measurement uncertainty of correction factors of fluconazole impurities determined by HPLC standard curve method / 中国药科大学学报

@#In this paper, the uncertainties of correction factors of fluconazole impurities determined by HPLC standard curve method were evaluated, and the main common factors affecting the accuracy of standard curve method were found, so as to improve the accuracy of the method.In this study, the corresponding fitting lines of fluconazole and its impurities A, B, C, D, F and I were established respectively, and the ratio of the slope of fitting lines of each impurity and its corresponding principal component was calculated as the correction factor of the impurity.Then on the basis of GUM method, the uncertainty of each impurity correction factor determined by standard curve method was evaluated according to the established uncertainty evaluation scheme of correction factor determination process.The correction factor and uncertainty of fluconazole impurities A, B, C, D, F and I were 1.068 ± 0.046, 0.102 ± 0.005, 0.0582 ± 0.0031, 1.382 ± 0.121, 0.802 ± 0.067 and 1.383 ± 0.119, respectively, and the coverage factor k was 2.Finally, the contribution rate of each uncertainty component was calculated.In the relative combined standard uncertainties urel(f) of fluconazole impurities A, B, C, D, F and I correction factors, the sum of contribution rate of slope uncertainty urel(K) of the linear equation of principal component and its impurity is more than 85%; in the slope uncertainties urel(K) of linear equation, the contribution rates of uncertainties of solution concentration in 8 of 12 data groups are more than 80%, and the contribution rates of uncertainties introduced by reference substance content in solution concentration are about 80%.It can be seen that the preparation of linear solution concentration is the most influential factor in the determination of impurity correction factor by standard curve method, followed by the linear fitting process.In the preparation process of linear solution concentration, the purity of reference substance is the most influential factor, followed by weighing and pipetting times.The conclusion can help the experimenters to better formulate experimental plans and ensure the accuracy of the results when doing similar work.

Analysis of resistance and sensitivity of 1 200 strains of vulvovaginal candidiasis in China to five common antifungal drugs / 中华妇产科杂志

Objective:To test the antibiotic susceptibility of vulvovaginal candidiasis pathogenic strains to 5 antifungal drugs commonly used in clinic.Methods:A total of 1 200 vulvovaginal candida patients from 23 gynecological and family planning outpatient departments in China were enrolled. Their vaginal secretions were collected for candida strain isolation and species identification. According to Clinical and Laboratory Standards Institute (CLSI) M27-S3, the sensitivity of 1 200 strains to clotrimazole, fluconazole, miconazole, itraconazole and nystatin was tested.Results:(1) The sensitivity and resistance of 1 200 vulvovaginal candidiasis pathogens to 5 antifungal drugs were statistically different ( χ2=3 513.201, P<0.01). (2) All strains had higher sensitivity to nystatin [99.92% (1 199/1 200)], followed by miconazole [92.25% (1 107/1 200)] and clotrimazole [87.17% (1 046/1 200)]. All strains had higher resistance to fluconazole [69.17% (830/1 200)], while itraconazole was 50.83% (610/1 200). (3) There was no significant difference between candida albicans and non-candida albicans in drug sensitivity to nystatin ( P=0.315) and miconazole ( P=0.425). (4) Candida albicans and non-candida albicans showed different sensitivity to clotrimazole, fluconazole and itraconazole, respectively. Compared with non-candida albicans, candida albicans showed higher sensitivity to clotrimazole [susceptibility rate: 73.01% (165/226) vs 90.45% (881/974); P<0.001] and higher resistance to fluconazole [resistance rate: 50.88% (115/226) vs 73.41% (715/974); P<0.001]. Although the drug sensitivity of itraconazole was not high, the susceptibility rate of candida albicans to itraconazole was slightly higher than that of non-candida albicans [37.68% (367/974) vs 23.89% (54/226)], and the drug resistance rate was lower [49.28% (480/974) vs 57.52% (130/226)]. Conclusions:The sensitivity of 1 200 strains of candida to 5 antifungal drugs is significantly different, the sensitivity rate of nystatin, miconazole and clotrimazole are higher, but the resistance rate of fluconazole and itraconazole are higher. The sensitivity of candida albicans and non-candida albicans to the same drug is also significantly different. It is suggested that in clinical diagnosis and treatment, we should pay attention to the identification of candida and drug sensitivity test, so as to select antifungal drugs rationally.

Antifungal activity of linalool against fluconazole-resistant clinical strains of vulvovaginal Candida albicans and its predictive mechanism of action

Candida albicans is the most frequently isolated opportunistic pathogen in the female genital tract, with 92.3% of cases in Brazil associated with vulvovaginal candidiasis (VVC). Linalool is a monoterpene compound from plants of the genera Cinnamomum, Coriandrum, Lavandula, and Citrus that has demonstrated a fungicidal effect on strains of Candida spp., but its mechanism of action is still unknown. For this purpose, broth microdilution techniques were applied, as well as molecular docking in a predictive manner for this mechanism. The main results of this study indicated that the C. albicans strains analyzed were resistant to fluconazole and sensitive to linalool at a dose of 256 µg/mL. Furthermore, the increase in the minimum inhibitory concentration (MIC) of linalool in the presence of sorbitol and ergosterol indicated that this molecule possibly affects the cell wall and plasma membrane integrity of C. albicans. Molecular docking of linalool with proteins that are key in the biosynthesis and maintenance of the cell wall and the fungal plasma membrane integrity demonstrated the possibility of linalool interacting with three important enzymes: 1,3-β-glucan synthase, lanosterol 14α-demethylase, and Δ 14-sterol reductase. In silico analysis showed that this monoterpene has theoretical but significant oral bioavailability, low toxic potential, and high similarity to pharmaceuticals. Therefore, the findings of this study indicated that linalool probably causes damage to the cell wall and plasma membrane of C. albicans, possibly by interaction with important enzymes involved in the biosynthesis of these fungal structures, in addition to presenting low in silico toxic potential.

Synergistic interaction of fluconazole/sodium bicarbonate on the inhibition of Candida glabrata phospholipase gene

Abstract Candida glabrata infections are responsible for deaths of people globally. Fluconazole is known to be less effective against C. glabrata, which developed many strategies to evade being destroyed by fluconazole. To achieve enhanced efficacy of fluconazole against C. glabrata, the interaction of fluconazole with sodium bicarbonate was investigated using the CLSI guidelines. The efficacy of fluconazole alone and in combination with sodium bicarbonate was evaluated using the time-kill and phospholipase production assays. Eventually, the expression of PLB was assessed using semi-quantitative RT-PCR to investigate the inhibitory properties of fluconazole alone and in combination with sodium bicarbonate against C. glabrata. The fluconazole/sodium bicarbonate combination displayed synergistic and antagonistic effects (FICI= 0.375-4.25). In C. glabrata ATCC, SN 152, and SN 164, the fluconazole/sodium bicarbonate combination exhibited a significant fungicidal activity (p< 0.05) but antagonistic effect in the case of SN 283. With exception of SN 283, a significant reduction was noted in phospholipase production in clinical isolates of C. glabrata treated with fluconazole/sodium bicarbonate combination. The PLB was down-regulated significantly by 0.168-0.515 fold in C. glabrata treated with fluconazole/sodium bicarbonate. The results suggested fluconazole/sodium bicarbonate to have a potential synergistic interaction in C. glabrata, and the underlying mechanism may be associated with phospholipase gene

Comparator product issues for biowaiver implementation: the case of Fluconazole

The aim of this work was to assess if the commercially available Fluconazole drug products (Reference, Generic and Similar) would meet the biowaiver criteria from Food and Drug Administration (FDA) and Brazilian Agency for Health Surveillance (ANVISA) agencies. All formulations were evaluated considering the dissolution profile carried out in Simulated Gastric Fluid (SGF) pH 1.2, Acetate Buffer (AB) pH 4.5 and Simulated Intestinal Fluid (SIF) pH 6.8. The results demonstrated that all formulations fulfilled the 85% of drug dissolved at 30 min criterion in SGF pH 1.2. However, in AB pH 4.5 and SIF pH 6.8, some formulations, including the comparator, did not achieve this dissolution percentage. The discrepant dissolution profiles also failed the ƒ2 similarity factor analysis, since none of the formulations showed values between 50 and 100 in the three dissolution media. Comparative dissolution profiles were not similar, considering that the main issues concerning the dissolution were evidenced for the comparator product. Hence, a revision in the regulatory norms in order to establish criteria to switch the comparator could result in an increased application of drugs based on biowaiver criteria

Criptococosis meníngea / Meningeal cryptococcosis

RESUMEN La criptococosis es una enfermedad producida por levaduras encapsuladas que se adquiere por la inhalación de propágulos infectantes de Cryptoccoccus, principalmente por la C. neoformnas y en menor frecuencia por la C. gatti. La distribución de este hongo es global, pero se encuentra de manera habitual en excretas de aves como las palomas. El principal compromiso en las personas es a nivel de los pulmones, del cerebro o de forma diseminada. La criptococosis en el sistema nervioso central (SNQ se presenta con meningoencefalitis, rara vez en forma de lesiones localizadas granulomatosas conocidas como criptococoma. Esta micosis es una causa frecuente de meningitis que se encuentra, especialmente, en los pacientes con VIH/SIDA. Las manifestaciones clínicas de esta enfermedad en el SNC son: cefalea, alteración del estado mental, fiebre, náuseas, vómito, deterioro visual, parálisis del sexto nervio craneal y signos de irritación meníngea, entre otras. El diagnóstico se realiza por medio de cultivo, microscopía del líquido cefalorraquídeo (LCR) o detección del antígeno de criptococo. El tratamiento de la meningitis por criptococo se divide en tres fases: inducción, consolidación y mantenimiento. Los pilares del tratamiento son la anfotericina B, la flucitosina y el fluconazol.

SUMMARY Cryptococcosis is a disease produced by encapsulated yeast that is acquired by inhalation of infecting Cryp-tococcus propagules, mainly by C. neoformnas and less frequently by C gatti. The distribution of this fungus is global, but it is commonly found in the excreta of birds such as pigeons. The main commitment in people is at the level of the lungs, the brain or in a disseminated way. Cryptococcosis in the central nervous system (CNS) presents with meningoencephalitis, rarely as localized granulomatous lesions known as cryptococcoma. This mycosis is a frequent cause of meningitis especially found in patients with HIV / AIDS. The clinical manifestations of cryptococcosis in the CNS are: headache, altered mental status, fever, nausea, vomiting, visual impairment, sixth cranial nerve palsy, and signs of meningeal irritation, among others. Diagnosis is made by culture, cerebrospinal fluid (CSF) microscopy, or by detection of cryptococcal antigen. The treatment of cryptococcal meningitis is divided into three phases: induction, consolidation, and maintenance. The mainstays of treatment are amphotericin B, flucytosine, and fluconazole.

Clinical analysis of 20 pediatric patients with kerion / 中华全科医师杂志

Twenty pediatric patients with kerion were treated in Department of Dermatology, Affiliated Hospital of Jining Medical University from January 2014 to June 2020. The general information, clinical manifestations, laboratory test results, treatment and prognosis were retrospectively analyzed. There were 13 males and 7 females aged from 2 to 10 years. Thirteen patients had a history of contact with animals, 4 had contact with parents with tinea. All patients had alopecia, 6 cases presented with inflammatory mass, 14 presented with abscessus; some patients had regional lymphadenopathy and febrile. Four cases were misdiagnosed as abscesses caused by bacterial infection and underwent incision leading to deep ulcers. A total of 13 fungal strains were isolated, including 4 strains of Microsporum gypseum, 3 strains of Trichophyton rubrum, 2 strains of Microsporum canis, the others were Trichophyton tonsurans and Trichophyton mentagrophytes and Fusarium. All patients were treated with fluconazole, concomitantly with topical antifungals and He-Ne laser, 19 of whom were cured. It is suggested that kerion characterized by inflammatory lesions is likely to be misdiagnosed. Fungal examination can confirm the diagnosis of kerion, and fluconazole is effective for treatment.

Clinical characteristics and influencing factors for mortality of patients with intra-abdominal candidiasis: a multicenter retrospective study / 中华消化外科杂志

Objective:To investigate the clinical characteristics and influencing factors of mortality in patients with intra-abdominal candidiasis (IAC).Methods:The retrospective case-control study was conducted. The clinicopathological data of 203 IAC patients who were admitted to 7 medical centers from June 2018 to June 2020 were collected, including 54 cases in Sir Run Run Shaw Hospital of Zhejiang University School of Medicine, 31 cases in Fujian Medical University Union Hospital, 25 cases in Beijing Hospital, 25 cases in the First Affiliated Hospital of Xi'an Jiaotong University, 24 cases in China-Japan Friendship Hospital, 22 cases in General Hospital of Eastern Theater Command of Chinese PLA and 22 cases in Chongqing University Cancer Hospital. There were 130 males and 73 females, aged (64±15)years. Observation indicators: (1) candida infection and treatment of IAC patients; (2) analysis of influencing factors for mortality of IAC patients. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was analyzed using the t test. Measurement data with skewed distribution were represented as M(range), and comparison between groups was analyzed using the Mann-Whitney U test. Count data were expressed as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Univariate and multivariate analyses were performed by Logistic regression model. Results:(1) Candida infection and treatment of IAC patients: 134 cases of candida albicans were cultured in the initial abdominal drainage fluid or intraoperative abdominal specimens of 203 patients, and 49 cases were treated with fluconazole. Of 69 cases infected with non candida albicans, 13 cases were treated with fluconazole. The resistance rate of candida albicans to fluconazole was 5.91%(12/203). Of 203 patients, there were 68 cases with infections shock, 53 cases with renal failure, 84 cases with respiratory failure and 63 cases with multiple organ failure, respectively. There were 148 of 203 patients admitted to intensive care unit for 9 days(range, 3-20 days), and the total hospital stay was 28 days(range, 17-50 days). Of 203 patients, 86 cases were cured and discharged, 50 cases were improved and transferred to local hospitals, 32 cases gave up treatment and discharged automatically, 19 cases died, 16 cases had no follow-up data. The mortality was 25.12%(51/203). (2) Analysis of influencing factors for mortality of IAC patients. Results of univariate analysis showed that acute physiology and chronic health evaluation score, sequential organ failure assessment score, the Cr, bilirubin, albumin, procalcitonin, and PLT on the first day of candida positive culture, of the lowest value in a week and the highest in a week, heart disease, diabetes, infections shock, renal failure, respiratory failure, multiple organ failure, anti-fungal therapy were the related factors for mortality of IAC patients ( t=-2.322, Z=-2.550, -2.262, -4.361, t=2.085, Z=-3.734, -5.226, -2.394, -5.542, t=3.462, Z=-4.957, -5.632, 3.670, -5.805, t=3.966, Z=-3.734, -5.727, χ2=4.071, 4.638, 27.353, 18.818, 13.199, 26.251, 13.388, P<0.05). Multivariate analysis showed that the bilirubin, procalcitonin on the first day of candida positive culture and infections shock were independent risk factors for mortality of IAC patients ( odds ratio=1.021, 1.022, 6.864, 95% confidence interval as 1.010-1.033, 1.001-1.044, 1.858-25.353, P<0.05). Conclusions:The common fungus of IAC was candida albicans, and fluconazole can be used as the initial empirical treatment. The prognosis of patients with abdominal candidiasis is poor. Bilirubin, procalcitonin on the first day of candida positive culture and infections shock are indepen-dent risk factors for mortality of IAC patients.

Study on the antifungal effect of amphotericin B combined with fluconazole at different time points / 口腔疾病防治

Objective@#To investigate the in vitro interaction of amphotericin B (AmB) and fluconazole (FLC) at different time points and provide a reference for clinical combined treatment therapy of polyenes and azoles.@*Methods@#Candida albicans ATCC SC5314 was used in the study. The minimum inhibitory concentration (MIC) of antifungal drugs was determined using the double microdilution broth method. The same amount of DMSO and low concentration drugs were added to the DMSO treatment group at different time points (0, 2, 4, 6 h) to determine whether the solvent background environment affected the growth of Candida albicans. In the experimental group, to observe the effect of low concentration AmB on the antifungal effect of FLC, the experimental group was administered a low concentration of AmB (0.25 μg/mL or 0.125 μg/mL) added to FLC at different time points (0, 2, 4, 6 h), and the same amount of DMSO was added to FLC at different time points in the single drug control group. In the experimental group, to observe the effect of low concentration of FLC on the antifungal effect of AmB, the experimental group was administered a low concentration of FLC (0.06 μg/mL or 0.03 μg/mL) in AmB at different time points (0, 2, 4, 6 h), and the same amount of DMSO was used at different time points as the single drug control group. In the solvent group, the same amounts of DMSO and low concentration drugs were added at different time points. After resuscitation, the colony growth of each solvent control group, single-drug control group and experimental group was observed to evaluate the interaction between drug concentration and time. Compared with the AmB single-drug control group, there was no significant change in the experimental group with added low concentrations of FLC at 0 h (F=0.27, P=0.775), which was 1.74-1.93 times that of the control group at 2-4 h (P < 0.001), and there was no significant difference in colony count after 6 h (P > 0.05). @*Results@# Under the treatment of FLC at an inhibitory concentration (0.25 μg/ml), adding low concentration AMB did not affect the antifungal effect of FLC, and the multiple of colony count differences were not significant (P > 0.05).@*Conclusion@#The interaction between AmB and FLC was time-dependent. At the early stage (0 h), the interaction effect between fluconazole and amphotericin B was not clear. The fungicidal effect of AmB could be weakened when FLC was supplied at 2-4 h, and the effect of FLC on AmB was absent after 6 h.

GC-MS analysis of low polarity extracts from Pinusmassoniana Lamb. leaves and study on their synergetic activity of fluconazole against fluconazole-resistant Candida albicans / 药学实践杂志

Objective To investigate the main chemical constituents of the low polarity extracts from pinusmassoniana Lamb. leaves and their synergetic activity with fluconazole against fluconazole-resistant Candida albicans. Methods The pinusmassoniana leaves were extracted with 80% ethanol, and then the extracts were extracted by petroleum ether to obtain the low polarity extracts. The chemical components were detected by GC-MS and elucidated by the comparison with the standard mass spectral data. The relative contents in percentage were calculated using the area normalization method. The minimal inhibitory concentrations (MIC80) of fluconazole-resistant Candida albicans strains by the low polarity extracts in combination with fluconazole were determined by checkerboard microdilution assay. Results 30 components were detected from the low polarity extracts, and 17 components were identified. The minimum inhibitory concentration (MIC80) of the 80% ethanol extracts, the low polarity extracts and the petroleum ether extracts from the pinusmassoniana leaves combined with fluconazole against fluconazole-resistant Candida albicans were 7.81 μg/ml, 31.25 μg/ml and >250 μg/ml, respectively. Conclusion The 80% ethanol extracts of pinusmassoniana leaves and its low polarity extracts have synergistic activity combined with fluconazole onfluconazole-resistant Candida albicans. The diterpenoids (53.99%) may be the effective components of the low polarity extracts.

Actividad antibacteriana y antifúngica del aceite esencial de Minthostachys mollis frente a microorganismos de la cavidad oral / Antibacterial and Antifungal Activity of the essential oil of Minthostachys mollis against oral microorganisms

Introducción: La Minthostachys mollises una planta aromática que crece en América Latina y produce aceites esenciales con acción antimicrobiana. Objetivo: Determinar la actividad del aceite esencial de Minthostachys mollis en diferentes concentraciones, comparado con doxiciclina y fluconazol frente a Porphyromonas gingivalis, Staphylococcus aureus y Candida albicans, a las 24, 48 y 72 horas. Métodos: Se realiza estudio experimental in vitro y longitudinal. Se prepararon 15 pocillos por subgrupo para evaluar el efecto inhibitorio de todas las concentraciones, dando un total de 360 pocillos. Por cromatografía de gases acoplada a espectrometría de masas se identificaron los componentes químicos del aceite esencial. Se analizó el efecto inhibitorio por el método de difusión de Kirby-Bauer en Agar Columbia y Agar Muller Hinton. El análisis estadístico se realizó mediante la prueba ANOVA y Tukey. Resultados: En el análisis químico se identificó principalmente pulegona (30,17 por ciento) y mentona (16,55 por ciento). Los halos de inhibición de Minthostachys mollis al 100 por ciento a las 24, 48 y 72 horas frente a la Porphyromonas gingivalis, midieron: 10,2 mm, 9,8 mm y 9,6 mm, respectivamente; frente al Staphylococcus aureus, midieron: 10,4 mm, 9,7 mm y 9,4 mm, respectivamente; y, por último, frente a Candida albicans midieron: 9,8 mm, 8,9 mm y 8,5 mm, respectivamente. Todas las concentraciones de Minthostachys mollis presentaron un efecto antimicrobiano significativamente menor que el fluconazol y la doxiciclina (p < 0,001). Conclusiones: El aceite esencial de Minthostachys mollis al 100 % presentó su mejor actividad inhibitoria frente al Staphylococcus aureus, la Porphyromonas gingivalis y la Candida albicans a las 24 horas. Sin embargo, este efecto antimicrobiano disminuye a medida que pasa el tiempo(AU)

Introduction: Minthostachys mollis is an aromatic plant species growing in Latin America which produces essential oils with antimicrobial activity. Objective: Determine the activity of essential oil from Minthostachys mollis at various concentrations as compared with doxycycline and fluconazole against Porphyromonas gingivalis, Staphylococcus aureus and Candida albicans at 24, 48 and 72 hours. Methods: An in vitro experimental longitudinal study was conducted. Fifteen wells were prepared per subgroup to evaluate the inhibitory effect of all concentrations, for a sum total of 360 wells. Chemical components of the essential oil were identified by gas chromatography-mass spectrometry. The inhibitory effect was analyzed with the Kirby-Bauer diffusion method in Mueller-Hinton and Columbia agar. Statistical analysis was based on ANOVA and Tukey's test. Results: Chemical analysis mainly found pulegone (30.17 percent) and menthone (16.55 percent). The inhibition halos of 100 percent Minthostachys mollis at 24, 48 and 72 hours against Porphyromonas gingivalis measured 10.2 mm, 9.8 mm and 9.6 mm, respectively, against Staphylococcus aureus they measured 10.4 mm, 9.7 mm and 9.4 mm, respectively, and against Candida albicans they measured 9.8 mm, 8.9 mm and 8.5 mm, respectively. The antimicrobial effect of Minthostachys mollis at all concentrations was significantly lower than that of fluconazole and doxycycline (p < 0.001). Conclusions: The essential oil from 100% Minthostachys mollis displayed its best inhibitory activity against Staphylococcus aureus, Porphyromonas gingivalis and Candida albicans at 24 hours. However, such antimicrobial effect decreases with the passing of time(AU)