Endophthalmitis Caused by Fusarium proliferatum

Fusarium proliferatum caused endophthalmitis after cataract surgery. Diagnosis was established by classical microbiology and molecular biology methods (PCR and DNA typing). The treatment with local amphotericin B, oral ketoconazole, and topical natamycin was successfu

Polymerase chain reaction diagnosis in fungal keratitis caused by Alternaria alternata

PURPOSE: To contribute toward assessing the effectiveness of polymerase chain reaction as a rapid method in diagnosis of torpid keratitis caused by opportunistic fungi. METHODS: Interventional case report. A 50-year-old man with a corneal abscess in the right eye treated for a period of 6 months with different combinations of broad-spectrum antibiotics and steroids was referred to our center. Corneal scraping was taken for microbiological study, including classic cultures and polymerase chain reaction. Amplified DNA was sequenced to identify the pathogen. RESULTS: Polymerase chain reaction amplification was negative for Acanthamoeba species and positive for fungi. The sequence analysis showed Alternaria alternata as the causal agent in 24 hours. Cultures confirmed the identification in 10 days. CONCLUSION: Polymerase chain reaction amplification with subsequent DNA-typing was revealed to be a useful method for detection of ocular pathogens such as A. alternata involved in cases of torpid keratitis, even in the presence of broad-spectrum antimicrobial therap

Rapid Molecular Diagnosis of Posttraumatic Keratitis and Endophthalmitis Caused by Alternaria infectoria

The first case of Alternaria infectoria ocular infection is reported. Keratitis and endophthalmitis developed after eye-perforating trauma from a lemon tree branch. Two months after surgery and empirical steroid and antibiotic treatment, diagnosis by molecular methods was performed. PCR amplification was positive for a fungus after 4 h. Antifungal treatment with amphotericin B and fluconazole was initiated immediately. DNA sequence analysis showed Alternaria infectoria to be the causal agent. After topical and systemic administration of antifungal treatment, ocular inflammation disappeared and visual acuity improved. DNA typing was found to be a useful tool to achieve early identification of the causal agen

Molecular epidemiology of isolates of the Cryptococcus neoformans species complex from Spain

6 p. : il.To study genetic diversity of Cryptococcus neoformans species complex in Spain, 97 isolates of the yeast recovered from human, animal and environmental samples have been analysed using three molecular epidemiological techniques. One of these, URA5 gene fragment length polymorphism (RFLP) analysis, has been previously described as a molecular epidemiology tool. Thus, standard profiles and reference strains have been defined for it. In addition, 5S rDNA/IGS RFLP and [GACA]4 microsatellite PCR fingerprinting were also used. Our results show five of the previously defined URA5 genotypes with a high frequency (33%) of the VNI type, which is in concordance with other studies. The high presence of VNIII pattern (28.9%) among our strains is remarkable and could be a specific feature of the isolates from our country. 5S rDNA/IGS RFLP showed a low intra-species discriminative power. Three different molecular profiles (S1-3), which showed a good correlation with the different species, varieties and genotypes, were obtained. [GACA]4 microsatellite PCR-fingerprinting analysis showed a high variability of patterns among the studied strains. Molecular profiles represented in a dendrogram clustered strains in four main groups related with the source of the yeast and also in concordance with some of the described genotypes (VNI-IV and VGI). ____________________________________________________________________________________.RESUMEN - Para ampliar el conocimiento de la diversidad genética que presenta el complejo Cryptococcus neoformans en España, se analizaron 97 aislamientos de esta levadura obtenidos a partir de muestras ambientales, veterinarias y clínicas, utilizando 3 técnicas epidemiológicas moleculares diferentes. Uno de estos marcadores moleculares, el análisis del URA5 RFLP, se ha descrito previamente como herramienta epidemiológica, por lo que se dispone de perfiles estándar y cepas de referencia. Además, se utilizaron también el análisis del RFLP del 5S rDNA/IGS y la huella digital tras amplificación por PCR de la secuencia microsatélite [GACA]4. Nuestros resultados muestran 5 de los genotipos URA5 con una elevada frecuencia del tipo VNI (33%), lo que concuerda con otros estudios. Es de resaltar la elevada presencia del perfil VNIII entre nuestras cepas (28,9%), lo que podría ser el rasgo específico más destacable de los aislamientos de nuestro país. El marcador 5S rDNA/IGS mostró un bajo poder discriminativo intraespecie. Se obtuvieron 3 perfiles moleculares distintos (S1-3), que presentaron una buena correlación con las especies, variedades y genotipos. La obtención de perfiles de huella digital con [GACA]4, presentó una gran variabilidad entre las cepas estudiadas. La representación en dendrograma agrupó las cepas en 4 agrupamientos principales relacionados con el origen de las levaduras y también con cierta concordancia con los genotipos descritos (VNI-IV y VGI)

Una halocina que actúa sobre el intercambiador Na+/H+ de Haloarchaea como un nuevo tipo de inhibidor de NHE de mamíferos

10 páginas, 3 figuras.[EN]: The capability of halocin H6 (a bacteriocin-like protein produced by haloarchaeaHaloferax gibbonsii) to inhibit Na+/H+ exchange (NHE) in mammalian cells and its cardio-protective efficacy on the ischemic and reperfused myocardium were evaluated in the present study. H6 inhibits NHE activity (measured by a flow cytometry method) in a dose-dependent form of cell lines of mammalian origin (HEK293, NIH3T3, Jurkat and HL-1) as well as in primary cell culture from human skeletal muscle (myocytes and fibroblasts).In vivo, an ischemia-reperfusion model in dogs by coronary arterial occlusion was used (two hours of regional ischemia and three hours of reperfusion). In animals treated with halocin H6 there was a significant reduction of premature ventricular ectopic beats and infarct size, whereas blood pressure and heart rate remained unchanged. Up to date, halocin H6 is the only described biological molecule that exerts a, specific inhibitory activity in NHE of eukaryotic cells.[ES]: En el presente trabajo se evalúa la capacidad de la halocina H6 (una proteína tipo bacteriocina producida por la haloarchaea Haloferax gibbonsii) para inhibir el intercambiador Na+/H+ (NHE) de células de mamífero y su posible eficacia cardioprotectora frente a los daños causados por isquemia-reperfusión del miocardio. En experimentos in vitro H6 inhibe la actividad de NHE (determinada por citometría de flujo) de forma dosis-dependiente tanto en líneas celulares de mamíferos (HEK293, NIH3T3, Jurkat y HL-1) como en cultivos primarios de miocitos y fibroblastos aislados de músculo esquelético humano. En experimentos in vivo se utilizó un modelo de isquemia- reperfusión en perros por oclusión de la arteria coronaria (dos horas de isquemia y tres de reperfusión). En animales tratados con halocina H6 se produjo una disminución significativa a nivel estadístico, tanto del número de latidos ectópicos ventriculares como del tamaño del infarto, mientras que no se produjeron cambios tanto en la presión sanguínea como en el ritmo cardíaco. Hasta la fecha la halocina H6 es la única molécula biológica descrita que ejerce una actividad inhibidora específica sobre el NHE de células eucariotas.M. Dolz held a fellowship of CSIC-Bancaja Foundation. This work was supported in part by Spanish Council for Scientific Research (CSIC- 2001551).Peer reviewe