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1.
Nutr Metab Cardiovasc Dis ; 28(4): 385-392, 2018 04.
Artículo en Inglés | MEDLINE | ID: mdl-29502926

RESUMEN

BACKGROUND AND AIMS: Phenylalanine (Phe) restricted diet, combined with Phe-free l-amino acid supplementation, is the mainstay of treatment for phenylketonuria (PKU). Being the diet a key factor modulating gut microbiota composition, the aim of the present paper was to compare dietary intakes, gut microbiota biodiversity and short chain fatty acids (SCFAs) production in children with PKU, on low-Phe diet, and in children with mild hyperphenylalaninemia (MHP), on unrestricted diet. METHODS AND RESULTS: We enrolled 21 PKU and 21 MHP children matched for gender, age and body mass index z-score. Dietary intakes, including glycemic index (GI) and glycemic load (GL), and fecal microbiota analyses, by means of denaturing gradient gel electrophoresis (DGGE) and Real-time PCR were assessed. Fecal SCFAs were quantified by gas chromatographic analysis. RESULTS: We observed an increased carbohydrate (% of total energy), fiber and vegetables intakes (g/day) in PKU compared with MHP children (p = 0.047), as well a higher daily GI and GL (maximum p < 0.001). Compared with MHP, PKU showed a lower degree of microbial diversity and a decrease in fecal butyrate content (p = 0.02). Accordingly, two of the most abundant butyrate-producing genera, Faecalibacterium spp. and Roseburia spp., were found significantly depleted in PKU children (p = 0.02 and p = 0.03, respectively). CONCLUSION: The low-Phe diet, characterized by a higher carbohydrate intake, increases GI and GL, resulting in a different quality of substrates for microbial fermentation. Further analyses, thoroughly evaluating microbial species altered by PKU diet are needed to better investigate gut microbiota in PKU children and to eventually pave the way for pre/probiotic supplementations.


Asunto(s)
Bacterias/metabolismo , Butiratos/metabolismo , Dieta con Restricción de Proteínas/efectos adversos , Microbioma Gastrointestinal , Tracto Gastrointestinal/microbiología , Fenilcetonurias/dietoterapia , Adolescente , Fenómenos Fisiológicos Nutricionales de los Adolescentes , Estudios de Casos y Controles , Niño , Fenómenos Fisiológicos Nutricionales Infantiles , Preescolar , Carbohidratos de la Dieta/administración & dosificación , Carbohidratos de la Dieta/metabolismo , Ácidos Grasos/metabolismo , Heces/química , Heces/microbiología , Fermentación , Índice Glucémico , Carga Glucémica , Humanos , Masculino , Estado Nutricional , Valor Nutritivo , Fenilcetonurias/sangre , Fenilcetonurias/microbiología , Fenilcetonurias/fisiopatología , Ingesta Diaria Recomendada , Resultado del Tratamiento
2.
Eur Rev Med Pharmacol Sci ; 18(5): 661-74, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24668706

RESUMEN

BACKGROUND: Candida bloodstream infections (BSI) represent an important problem in Intensive Care Units (ICUs). The epidemiology of candidemia is changing with an increase in the proportion of Candida (C.) non-albicans. OBJECTIVES: An Italian 2-year observational survey on ICU was conducted to evaluate the species distribution and possible differences between BSI caused by C. albicans and C. non-albicans. For comparative purposes, we performed a European literature-based review to evaluate distribution and frequency of Candida spp. causing ICU candidemia, during the period 2000-2013. MATERIALS AND METHODS: This laboratory-based survey involved 15 microbiology centers (GISIA-3 study). All candidemia episodes in adult patients were considered. Data were prospectively collected from 2007 to 2008. PubMed was searched for peer-reviewed articles. RESULTS: In total, 462 candidemia episodes were collected. C. albicans accounted for 49.4% of the isolates, followed by C. parapsilosis (26.2%) and C. glabrata (10.4%). Mortality was higher in patients with C. non-albicans than C. albicans (47.3% vs. 32.4 %, p > 0.05). Among risk factors, parenteral nutrition was more common (p = 0.02) in non-albicans candidemia, while surgery was more frequent (p = 0.02) in C. albicans candidemia. Twenty-four relevant articles were identified. C. albicans was the predominant species in almost all studies (range 37.9% -76.3%). C. glabrata was commonly isolated in the German-speaking countries, France, UK and North Europe; C. parapsilosis in Turkey, Greece and Spain. CONCLUSIONS: Although C. non-albicans BSI is increasing, our study shows that C. albicans is still the predominant species in ICU candidemia. There are differences in the epidemiology of Candida BSI among European countries, with a prevalence of C. glabrata and C. parapsilosis in Northern and Southern countries, respectively.


Asunto(s)
Candidemia/diagnóstico , Candidemia/epidemiología , Unidades de Cuidados Intensivos/tendencias , Adulto , Antifúngicos/uso terapéutico , Candidemia/tratamiento farmacológico , Europa (Continente)/epidemiología , Francia/epidemiología , Grecia/epidemiología , Humanos , Persona de Mediana Edad , Estudios Observacionales como Asunto/métodos , Estudios Prospectivos , Factores de Riesgo , España/epidemiología , Encuestas y Cuestionarios , Turquía/epidemiología
3.
J Biol Regul Homeost Agents ; 28(4): 743-52, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-25620183

RESUMEN

The aim of this study was to investigate the effects of different species of Lactobacilli on hyphal formation and biofilm development by the opportunistic fungal pathogen Candida albicans. We employed 4 different Lactobacillus species, namely L. rhamnosus, L. acidophilus, L. plantarum and L. reuteri, and 2 C. albicans strains, the reference DAY286 and its isogenic hwp1/hwp1 mutant, the FJS24 strain. As assessed by morphological analysis and quantitative colorimetric assays, Lactobacillus crude filtrate supernatant fluids (CFSF) affected Candida, impairing both hyphal formation and biofilm production. The CFSF-mediated phenomenon occurred in a dilution- and time-dependent manner and was consistently observed, irrespective of the C. albicans HWP1 genotype.


Asunto(s)
Biopelículas/crecimiento & desarrollo , Candida albicans/fisiología , Hifa/crecimiento & desarrollo , Lactobacillus
4.
Mycopathologia ; 173(2-3): 135-8, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21997860

RESUMEN

We report a case of a 40-year-old Caucasian woman who came under our observation with a 7-year history of a chronic erythematous scaly dermatitis, diagnosed as psoriasis, involving gluteal area and thighs, and treated with topical steroids without benefit. During pregnancy, a progressive worsening of her condition and an extension of cutaneous lesions were observed. Her newborn, a 15-day-old girl, presented a similar scaly and squamous lesion on her scalp. Mycological examination was positive for Trichophyton rubrum in both cases, and random amplified polymorphic DNA analysis confirmed the isogenicity of the two isolates. We performed a diagnosis of T. rubrum tinea corporis and tinea capitis. The case we describe illustrates an unusual clinical presentation of tinea corporis with remarkable extension of cutaneous lesions due to the diagnostic delay and the continuous use of local steroids, together with a rare tinea capitis in the newborn. Our experience highlights the possibility of mother-child transmission and the importance of an early diagnosis.


Asunto(s)
Transmisión Vertical de Enfermedad Infecciosa , Complicaciones Infecciosas del Embarazo/microbiología , Tiña del Cuero Cabelludo/transmisión , Tiña/transmisión , Trichophyton/aislamiento & purificación , Adulto , Antifúngicos/uso terapéutico , Femenino , Humanos , Recién Nacido , Embarazo , Tiña/diagnóstico , Tiña/tratamiento farmacológico , Tiña/microbiología , Tiña del Cuero Cabelludo/diagnóstico , Tiña del Cuero Cabelludo/tratamiento farmacológico , Tiña del Cuero Cabelludo/microbiología , Trichophyton/genética , Trichophyton/fisiología
5.
Eur J Clin Microbiol Infect Dis ; 30(11): 1437-41, 2011 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-21479840

RESUMEN

In the present work, we studied the distribution of Candida parapsilosis complex species and the antifungal susceptibility of clinical isolates collected during an Italian surveillance study of yeast invasive fungal infections (IFIs) in intensive care units (ICUs). Minimum inhibitory concentrations (MICs) were determined using the Clinical and Laboratory Standards Institute (CLSI) reference broth microdilution method. BanI digestion patterns of the secondary alcohol dehydrogenase polymerase chain reaction (PCR) products were used to identify C. parapsilosis sensu stricto, C. orthopsilosis, and C. metapsilosis. A total of 138 C. parapsilosis isolates were stored (January 2007-December 2008). The overall frequency of C. parapsilosis complex in IFIs was 22%. Of the 138 tested isolates, 95% were C. parapsilosis sensu stricto, 3.6% were C. orthopsilosis, and 1.4% were C. metapsilosis. The MIC(50) values (expressed as µg/ml) for anidulafungin, caspofungin, and micafungin for C. parapsilosis complex were 2, 1, and 2, respectively, and the MIC(90) values were 4, 2, and 4, respectively. The MIC(50) and MIC(90) values for itraconazole and posaconazole were 0.12 and 0.25, respectively, and for fluconazole, they were 1 and 4, respectively. This study, the most comprehensive study conducted to date to evaluate the frequency and antifungal susceptibility profiles of C. parapsilosis complex isolates from critically ill patients in Italy, highlights the low prevalence of C. orthopsilosis and C. metapsilosis in IFIs.


Asunto(s)
Alcohol Deshidrogenasa/genética , Antifúngicos/farmacología , Candida/clasificación , Candidiasis Invasiva/epidemiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Anfotericina B/farmacología , Candida/efectos de los fármacos , Candida/genética , Candida/aislamiento & purificación , Candidiasis Invasiva/microbiología , Niño , Preescolar , Enfermedad Crítica , Equinocandinas/farmacología , Proteínas Fúngicas/genética , Humanos , Lactante , Recién Nacido , Unidades de Cuidados Intensivos , Italia/epidemiología , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Técnicas de Tipificación Micológica , Reacción en Cadena de la Polimerasa , Triazoles/farmacología , Adulto Joven
7.
Minerva Anestesiol ; 76(11): 950-6, 2010 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-21102391

RESUMEN

Invasive fungal infections (IFIs) are on the increase not only among oncology and transplant patients but also among patients admitted to intensive care units (ICU). The rise in ICU IFIs can be attributed to the growing use of complex surgical procedures, invasive medical devices, and long-term, broad-spectrum antibiotic therapy. The majority of these life-threatening infections are caused by the well-known opportunistic pathogens Candida albicans and Aspergillus fumigatus, but new opportunistic pathogens, including yeast-like and other filamentous fungi, have emerged as additional causes. Invasive Candida infections, particularly candidemia, represent the most common IFI in critically ill patients. The species that cause candidemia markedly differ in their responses to antifungal drugs; for this reason, therapy must be tailored to the susceptibility characteristics of the infectious agent. Candidemia caused by non-albicans Candida species is increasing worldwide, and these infections are generally associated with high mortality rates, particularly bloodstream infections caused by C. krusei, which is innately resistant to fluconazole, or C. glabrata, which easily develops azole resistance. Although invasive yeast infections can be considered the most important causes of morbidity and mortality in ICU patients, pulmonary aspergillosis has recently emerged as an additional complication. Diagnosis of IFIs can be achieved using conventional approaches (microscopy, culture, and serology) and newer methods, including antigen detection and polymerase chain reaction (PCR) assays. Because most of the conventional approaches lack sensitivity, antigen detection and PCR assays could represent a valid alternative; however, these procedures need to be standardized and evaluated in a large number of patients.


Asunto(s)
Infección Hospitalaria/diagnóstico , Infección Hospitalaria/epidemiología , Unidades de Cuidados Intensivos , Micosis/diagnóstico , Micosis/epidemiología , Infección Hospitalaria/microbiología , Infección Hospitalaria/terapia , Humanos , Micosis/microbiología , Micosis/terapia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
8.
J Chemother ; 21(3): 322-9, 2009 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-19567354

RESUMEN

The aims of the study were to analyze the clinical and epidemiological characteristics and treatments for patients who developed zygomycosis enrolled in Italy during the European Confederation of Medical Mycology of medical mycology survey. This prospective multicenter study was performed between 2004 and 2007 at 49 italian Departments. 60 cases of zygomycosis were enrolled: the median age was 59.5 years (range 1-87), with a prevalence of males (70%). The majority of cases were immunocompromised patients (42 cases, 70%), mainly hematological malignancies (37). Among non-immunocompromised (18 cases, 30%), the main category was represented by patients with penetrating trauma (7/18, 39%). The most common sites of infection were sinus (35%) with/without CNS involvement, lung alone (25%), skin (20%), but in 11 cases (18%) dissemination was observed. According to EORTC criteria, the diagnosis of zygomycosis was proven in 46 patients (77%) and in most of them it was made in vivo (40/46 patients, 87%); in the remaining 14 cases (23%) the diagnosis was probable. 51 patients received antifungal therapy and in 30 of them surgical debridement was also performed. The most commonly used antifungal drug was liposomal amphotericin B (L-AmB), administered in 44 patients: 36 of these patients (82%) responded to therapy. Altogether an attributable mortality rate of 32% (19/60) was registered, which was reduced to 18% in patients treated with L-AmB (8/44). Zygomycosis is a rare and aggressive filamentous fungal infection, still associated with a high mortality rate. This study indicates an inversion of this trend, with a better prognosis and significantly lower mortality than that reported in the literature. It is possible that new extensive, aggressive diagnostic and therapeutic procedures, such as the use of L-AmB and surgery, have improved the prognosis of these patients.


Asunto(s)
Cigomicosis/epidemiología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Niño , Preescolar , Farmacorresistencia Fúngica , Femenino , Humanos , Huésped Inmunocomprometido , Lactante , Italia/epidemiología , Masculino , Persona de Mediana Edad , Cigomicosis/diagnóstico , Cigomicosis/tratamiento farmacológico , Cigomicosis/etiología
9.
J Chemother ; 19(5): 508-13, 2007 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-18073149

RESUMEN

We evaluated the in vitro activity of voriconazole, amphotericin B, and itraconazole against 192 clinical mould isolates recovered in twenty Italian microbiology laboratories. The vast majority of isolates belonged to the genus Aspergillus (94.2%) with A. fumigatus (58.3%) being the most frequently isolated species. Antifungal susceptibility testing was performed using the broth microdilution method defined by the CLSI M38-A standard, and results were compared to those obtained with Sensititre panels. Aspergillus flavus ATCC 204304 was employed as reference strain and results were within all expected ranges. Voriconazole's activity against the 192 mould isolates was comparable to that of amphotericin B and itraconazole: voriconazole MIC90 (CLSI 1 microg/ml, Sensititre 1 microg/ml), itraconazole MIC90 (CLSI 0.5 microg/ml, Sensititre 0.5 microg/ml), amphotericin B MIC90 (CLSI 1 microg/ml, Sensititre 1 microg/ml). In conclusion, these in vitro data highlight voriconazole's broad spectrum activity against filamentous fungi and support its use as a first line agent for the treatment of fungal diseases.


Asunto(s)
Anfotericina B/farmacología , Antifúngicos/farmacología , Farmacorresistencia Fúngica , Hongos/efectos de los fármacos , Itraconazol/farmacología , Pirimidinas/farmacología , Triazoles/farmacología , Humanos , Técnicas In Vitro , Pruebas de Sensibilidad Microbiana , Micosis/tratamiento farmacológico , Voriconazol
10.
J Chemother ; 16(5): 474-8, 2004 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-15565915

RESUMEN

Voriconazole, amphotericin B and itraconazole were tested in vitro against 18 strains of Aspergillus fumigatus isolated from cystic fibrosis patients. Susceptibility was tested with the broth microdilution method (M38-A protocol-NCCLS). Results of this reference method were compared with those of an experimental commercial microdilution broth method (Sensititre). Two different inocula, prepared from 2- and 7-day cultures, were used. Minimum inhibitory concentrations (MICs) of the reference method ranged from 0.25 to 2 microg/ml for voriconazole, 0.06 to 1 microg/ml for amphotericin B, 0.016 to >16 microg/ml for itraconazole. There were no significant differences in the MIC ranges or MIC90 values obtained with the two testing methods or with the two types of inocula. These findings confirm the good in vitro activity of voriconazole, itraconazole and amphotericin B against A. fumigatus. They also indicate that reliable susceptibility data can be generated more rapidly by commercial systems and use of 2-day cultures for inoculum preparation.


Asunto(s)
Antifúngicos/farmacología , Aspergillus fumigatus/efectos de los fármacos , Pruebas de Sensibilidad Microbiana/métodos , Anfotericina B/farmacología , Humanos , Itraconazol/farmacología , Valor Predictivo de las Pruebas , Pirimidinas/farmacología , Sensibilidad y Especificidad , Triazoles/farmacología , Voriconazol
11.
Eur J Clin Microbiol Infect Dis ; 23(8): 619-24, 2004 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-15258831

RESUMEN

The antifungal susceptibility of 309 Candida glabrata and 63 Candida krusei clinical isolates was tested via the Sensititre YeastOne-3 system (Trek Diagnostic Systems, East Grinstead, UK) to compare the in vitro activity of voriconazole with that of five other antifungal agents (amphotericin B, fluconazole, itraconazole, ketoconazole, and flucytosine). Voriconazole was highly active (MIC90, 0.5 microg/ml) against isolates of both species, including those for which the MICs of itraconazole and fluconazole were high (MIC90s of itraconazole, 2 microg/ml for C. glabrata and 0.5 microg/ml for C. krusei; MIC90s of fluconazole, 32 microg/ml for C. glabrata and 64 microg/ml for C. krusei). Ketoconazole MIC90 values for both species were identical to those of voriconazole. The MIC90 of amphotericin B was similar for both species (0.125 microg/ml for C. glabrata and 0.25 microg/ml for C. krusei). As expected, flucytosine was only moderately active against C. krusei isolates (MIC90, 16 microg/ml) but was highly active against C. glabrata isolates (MIC90, 0.03 microg/ml). Potential cross-resistance within the azole class was noted for some strains of C. glabrata (5.5%) that presented high MIC values for all the azoles tested. In order to consider voriconazole a viable alternative to other triazoles for the treatment of infections caused by Candida species, susceptibility testing of all clinically significant isolates of C. glabrata and C. krusei is recommended because of the potential for azole cross-resistance. The Sensititre YeastOne-3 seems to be a suitable commercial tool for this purpose.


Asunto(s)
Antifúngicos/farmacología , Candida glabrata/efectos de los fármacos , Candida glabrata/aislamiento & purificación , Farmacorresistencia Fúngica , Pirimidinas/farmacología , Triazoles/farmacología , Candida/clasificación , Candida/efectos de los fármacos , Candidiasis/diagnóstico , Candidiasis/tratamiento farmacológico , Humanos , Técnicas In Vitro , Italia , Pruebas de Sensibilidad Microbiana , Muestreo , Sensibilidad y Especificidad , Voriconazol
12.
J Clin Microbiol ; 40(8): 2953-8, 2002 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-12149358

RESUMEN

Fluconazole susceptibility among 800 clinical Candida isolates (60% C. albicans) and two control strains (C. krusei ATCC 6258 and C. parapsilosis ATCC 22019) was tested with the NCCLS M27-A method (gold standard) and six commercial products (Candifast, disk, Etest, Fungitest, Integral System Yeasts, and Sensititre YeastOne). Results were classified as susceptible, susceptible-dose dependent, or resistant using M27-A breakpoints or, for Fungitest, Integral System Yeasts, and Candifast, as susceptible, intermediate, or resistant, according to the manufacturers' instructions. Concordance with NCCLS M27-A results was analyzed with the chi(2) test. Intra- and interlaboratory reproducibility was also evaluated. NCCLS M27-A (90.1%), Etest (93.1%), Sensititre YeastOne (93.1%), disk (96.7%), Fungitest (92.6%), Integral System Yeasts (40.6%), and Candifast (6.0%) classified the indicated percentages of C. albicans isolates as susceptible. Among non-C. albicans strains, the percentages of susceptible isolates were as follows: NCCLS M27-A, 74.0%; Etest, 83.8%; Sensititre YeastOne, 64.1%; disk, 60.6%; Fungitest, 76.6%; Integral System Yeasts, 28.3%; and Candifast, 27.4%. All methods except Candifast and Integral System Yeasts showed good agreement with NCCLS M27-A results for both C albicans and non-C. albicans isolates. Intralaboratory reproducibility was excellent for NCCLS M27-A, Etest, Sensititre YeastOne, disk, and Fungitest (88 to 91%). Similar results emerged from the interlaboratory reproducibility evaluation. Our findings indicate that some commercial methods can be useful for fluconazole susceptibility testing of clinical Candida isolates. Those characterized by a lack of medium standardization and/or objective interpretative criteria should be avoided. Particular caution is necessary when testing is being done for clinical and epidemiological purposes.


Asunto(s)
Antifúngicos/farmacología , Candida/efectos de los fármacos , Fluconazol/farmacología , Pruebas de Sensibilidad Microbiana/normas , Juego de Reactivos para Diagnóstico , Candidiasis/microbiología , Humanos , Laboratorios , Pruebas de Sensibilidad Microbiana/métodos , Reproducibilidad de los Resultados
13.
J Appl Microbiol ; 92(5): 828-36, 2002.
Artículo en Inglés | MEDLINE | ID: mdl-11972685

RESUMEN

AIMS: The persistent circulation of hepatitis A virus (HAV) in the Mediterranean area suggests the need for monitoring its presence in the environment. A reverse transcriptase-polymerase chain reaction (RT-PCR) was used to detect the presence of HAV in several consecutive raw sewage and final effluent samples, collected over an 8-month period from an activated sludge treatment plant in southern Italy. METHODS AND RESULTS: Two distinct purification protocols, either based on antigen-capture with monoclonal antibody (AC) or RNA extraction, were compared. The possible influence of the antibody used in the AC phase was evaluated in preliminary experiments on HAV-spiked samples, using two different monoclonal antibodies. Hepatitis A virus RNA was detected in all but one sewage environmental sample examined. The contemporary presence of enteroviruses, reoviruses and phages was observed, while HAV growth in cell culture was hampered. CONCLUSIONS: The RT-PCR technique was confirmed to be a valuable tool for the rapid monitoring of HAV in sewage samples. In addition, this study demonstrated that application of different sample purification methods can result in different levels of sensitivity of the assay and that, in the antigen-capture method, the choice of antibody can have a crucial role. SIGNIFICANCE AND IMPACT OF THE STUDY: This work underlines the need for technical uniformity in environmental studies from different laboratories for a correct and useful comparison of the results.


Asunto(s)
Virus de la Hepatitis A/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Aguas del Alcantarillado/microbiología , Microbiología del Agua , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Antígenos Virales/química , Antígenos Virales/inmunología , Antígenos Virales/aislamiento & purificación , Genoma Viral , Virus de la Hepatitis A/química , Virus de la Hepatitis A/genética , Humanos , Datos de Secuencia Molecular , ARN Viral/análisis , ARN Viral/aislamiento & purificación , Sensibilidad y Especificidad , Análisis de Secuencia de ADN , Proteínas Virales/química
14.
J Pediatr Gastroenterol Nutr ; 33(5): 629-32, 2001 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-11740244

RESUMEN

BACKGROUND: The recently discovered TT virus (TTV) has been shown to be highly prevalent in patients with cryptogenetic chronic liver disease and fulminant hepatitis. To study the frequency of TTV and to evaluate the possible association with liver disease, 37 pediatric and young adult patients with thalassemia, and 36 healthy children were included in the study. The sera of 100 blood donors selected randomly in the same period were also tested for TTV DNA. METHODS: The TTV amplification by polymerase chain reaction (PCR) was performed using a first set of primers that recognize an internal sequence into N22 and a second set of primers amplifying a sequence within 5;NCR (5; noncoding region). RESULTS: The first set of primers revealed TTV DNA in 73% of thalassemic patients, in 8% of healthy children, and in 5% of healthy blood donors. With the second set of primers, the prevalence of TTV DNA was, respectively, 100% in thalassemic patients, 44.5% in healthy pediatric patients, and 87% in healthy blood donors. All individuals who tested positive for TTV by the first set of primers were also positive by the second primer set. The TTV infection seemed not to be the cause of altered transaminase levels. Sequencing of TTV clones from thalassemic patients showed the presence of different TTV variants in the same serum. CONCLUSION: The prevalence of TTV in polytransfused children is similar to that detected in blood donors. Moreover, TTV can be detected in healthy children of all ages. The presence of TTV seems to have no clinical significance.


Asunto(s)
Infecciones por Virus ADN/epidemiología , Talasemia/complicaciones , Torque teno virus/aislamiento & purificación , Adolescente , Adulto , Secuencia de Bases , Donantes de Sangre , Niño , Preescolar , Infecciones por Virus ADN/sangre , ADN Viral/análisis , ADN Viral/química , Femenino , Humanos , Lactante , Masculino , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Prevalencia , Análisis de Secuencia de ADN , Talasemia/sangre , Talasemia/virología , Torque teno virus/genética , Torque teno virus/patogenicidad
15.
Ital Heart J Suppl ; 1(11): 1464-71, 2000 Nov.
Artículo en Italiano | MEDLINE | ID: mdl-11109197

RESUMEN

BACKGROUND: Evoked ischemia induces morphological modifications in QRS complex that can be detected by a new electrocardiographic computerized system that we have developed (ventricular ischemic site analysis-VISA). The aim of this study was to evaluate, by continuous recording, the ischemia evolution during stress or pharmacological test and to correlate the modifications of QRS with the shift of the ST-T segment. METHODS: According to our method, the vectorial differences between rest QRS and stress QRS are recorded as deformations of the QR and/or RS segments. Sixty patients were studied: 44 had inducible ischemia during single photon emission computed tomography 99mTc-sestamibi test; 33 of these underwent ergometric test and 11 pharmacological test. RESULTS: The VISA test was positive in 91% of patients against the 61% with standard ECG. Ischemic deformations of the QRS appeared at 62.1% of stress duration, while the ST-T shift appeared at 81.4% of the test. During pharmacological stress the ST-T segment did not change. CONCLUSIONS: Our method is more sensitive than standard ECG in identifying inducible ischemia. Ischemic deformations of the QRS complex occur earlier than the repolarization modifications and they may be present also in their absence. The higher sensitivity of the VISA test depends on the fact that QRS ischemic modifications are caused both by the lesion current and a conduction delay in the ischemic zone.


Asunto(s)
Diagnóstico por Computador , Electrocardiografía , Isquemia Miocárdica/diagnóstico , Isquemia Miocárdica/fisiopatología , Humanos , Vectorcardiografía
16.
Diagn Microbiol Infect Dis ; 38(1): 29-36, 2000 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-11025181

RESUMEN

Fluconazole susceptibility was tested in 100 clinical yeast isolates (65 Candida albicans, 13 C. glabrata, 8 C. tropicalis, 7 C. parapsilosis, 3 Saccharomyces cerevisiae, 1 each of C. krusei, C. lusitaniae, Cryptococcus neoformans, Rhodotorula glutinis) and two control strains (Candida krusei ATCC 6258, C. parapsilosis ATCC 22019) using broth microdilution (reference method), disk diffusion, Etest strips, Sensititre YeastOne, Candifast, Integral System Yeasts. Using M27-A breakpoints, isolates were classified as susceptible (81%), susceptible-dose dependent or Resistant with broth dilution. Rates of concordance with the reference method were good for Sensititre YeastOne, Etest and disc-diffusion (81.2%-94.7%) but very low for the Candifast (3.1%) and Integral System (16.6%), which classified most susceptible isolates as resistant. Lack of standardisation (inoculum, medium composition) and non-objective interpretation schemes may be the cause of their poor performance. Sensititre YeastOne, Etest and disc-diffusion are potentially useful for fluconazole antifungal susceptibility testing of yeasts in clinical laboratories.


Asunto(s)
Antifúngicos/farmacología , Fluconazol/farmacología , Levaduras/efectos de los fármacos , Candida/efectos de los fármacos , Cryptococcus neoformans/efectos de los fármacos , Farmacorresistencia Microbiana , Humanos , Pruebas de Sensibilidad Microbiana , Reproducibilidad de los Resultados , Rhodotorula/efectos de los fármacos , Saccharomyces cerevisiae/efectos de los fármacos
17.
J Clin Microbiol ; 38(4): 1609-14, 2000 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-10747151

RESUMEN

A PCR-based assay was developed to detect and identify medically important yeasts in clinical samples. Using a previously described set of primers (G. Morace et al., J. Clin. Microbiol. 35:667-672, 1997), we amplified a fragment of the ERG11 gene for cytochrome P-450 lanosterol 14alpha-demethylase, a crucial enzyme in the biosynthesis of ergosterol. The PCR product was analyzed in a reverse cross blot hybridization assay with species-specific probes directed to a target region of the ERG11 gene of Candida albicans (pCal), C. guilliermondii (pGui), C. (Torulopsis) glabrata (pGla), C. kefyr (pKef), C. krusei (pKru), C. parapsilosis (pPar), C. tropicalis (pTro), the newly described species C. dubliniensis (pDub), Saccharomyces cerevisiae (pSce), and Cryptococcus neoformans (pCry). The PCR-reverse cross blot hybridization assay correctly identified multiple isolates of each species tested. No cross-hybridization was detected with any other fungal, bacteria, or human DNAs tested. The method was tested against conventional identification on 140 different clinical samples, including blood and cerebrospinal fluid, from patients with suspected fungal infections. The results agreed with those of culture and phenotyping for all but six specimens (two of which grew yeasts not included in the PCR panel of probes and four in which PCR positivity-culture negativity was justified by clinical findings). Species identification time was reduced from a mean of 4 days with conventional identification to 7 h with the molecular method. The PCR-reverse cross blot hybridization assay is a rapid method for the direct detection and identification of yeasts in clinical samples.


Asunto(s)
Candida/aislamiento & purificación , Cryptococcus neoformans/aislamiento & purificación , Sistema Enzimático del Citocromo P-450/genética , Hibridación de Ácido Nucleico/métodos , Oxidorreductasas/genética , Reacción en Cadena de la Polimerasa/métodos , Saccharomyces cerevisiae/aislamiento & purificación , Candida/clasificación , Candida/enzimología , Candidiasis/microbiología , Criptococosis/microbiología , Cryptococcus neoformans/clasificación , Cryptococcus neoformans/enzimología , ADN de Hongos/análisis , ADN de Hongos/genética , Humanos , Datos de Secuencia Molecular , Técnicas de Tipificación Micológica , Sondas de Oligonucleótidos , Mapeo Restrictivo , Saccharomyces cerevisiae/clasificación , Saccharomyces cerevisiae/enzimología , Especificidad de la Especie , Esterol 14-Desmetilasa
18.
J Hosp Infect ; 44(3): 186-92, 2000 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-10706801

RESUMEN

Technical complications and nosocomial bloodstream infections associated with short-term central venous catheterization remain a heavy burden in terms of morbidity, mortality and cost in HIV-positive subjects. Between 1994 and 1997, 327 central venous catheters (CVCs) inserted in 212 patients for a total of 5005 catheter days were investigated. Forty-two technical complications (13%) occurred in 40 patients. Logistic regression analysis revealed that a high APACHE III score was associated with development of CVC-related complications (P = 0.01). One hundred and eight of 327 CVCs (33%) were suspected as being infected. However only 61 episodes (61/327, 19%) were finally diagnosed as CVC-related sepsis. Three variables affecting the rate of CVC-related sepsis were identified: 1) administration of TPN (P = 0.01); 2) low number of circulating CD4+ cells (P = 0.04); 3) high APACHE III score (P = 0. 04). Doctors responsible for AIDS patients should carefully consider the relative risks and benefits of CVC insertion in an individual patient.


Asunto(s)
Infecciones Oportunistas Relacionadas con el SIDA/etiología , Cateterismo Venoso Central/efectos adversos , Infección Hospitalaria/etiología , Morbilidad , Sepsis/etiología , Infecciones Oportunistas Relacionadas con el SIDA/diagnóstico , Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Infecciones Oportunistas Relacionadas con el SIDA/mortalidad , APACHE , Adulto , Recuento de Linfocito CD4 , Infección Hospitalaria/diagnóstico , Infección Hospitalaria/microbiología , Infección Hospitalaria/mortalidad , Falla de Equipo , Femenino , Hospitales Universitarios , Humanos , Control de Infecciones/métodos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Nutrición Parenteral Total/efectos adversos , Estudios Prospectivos , Factores de Riesgo , Ciudad de Roma/epidemiología , Sepsis/diagnóstico , Sepsis/microbiología , Sepsis/mortalidad
19.
Virology ; 264(2): 410-21, 1999 Nov 25.
Artículo en Inglés | MEDLINE | ID: mdl-10562502

RESUMEN

The small hydrophobic protein 3AB of the picornaviruses, encompassing the replication primer 3B, has been suggested to anchor the viral replication complex to membranes. For hepatitis A virus (HAV) 3AB, we have previously demonstrated its ability to form stable homodimers, to bind to membranes, and to interact specifically with RNA, implicating its multiple involvement in viral replication. In the present report, we show that HAV 3AB additionally interacts with HAV protein 3CD, a feature also described for the corresponding polypeptide of poliovirus. By assessing the interactions of three deletion mutants, distinct domains of HAV 3AB were mapped. The hydrophobic domain and the 3B moiety were found to be essential for the 3AB interaction with 3CD. Both electrostatic and hydrophobic forces are involved in this interaction. The cluster of charged amino acid residues at the C terminus of 3A seems to determine the specificity of 3AB interaction with RNA structures formed at either terminus of the HAV genome. Furthermore, our data implicate that 3A can interact with HAV RNA. Compared with poliovirus 3AB, which by itself is a nonspecific RNA-binding protein, HAV 3AB specifically recognizes HAV RNA structures that might be of relevance for initiation of viral RNA replication.


Asunto(s)
Cisteína Endopeptidasas/metabolismo , Hepatovirus/fisiología , ARN Viral/metabolismo , Proteínas del Núcleo Viral/metabolismo , Proteínas Virales , Proteasas Virales 3C , Secuencia de Aminoácidos , Sitios de Unión , Mapeo Cromosómico , Cisteína Endopeptidasas/genética , Dimerización , Hepatovirus/genética , Hepatovirus/metabolismo , Humanos , Datos de Secuencia Molecular , Homología de Secuencia de Aminoácido , Proteínas del Núcleo Viral/genética
20.
Minerva Anestesiol ; 65(9): 637-9, 1999 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-10522134

RESUMEN

BACKGROUND: It has been observed that the incidence and prevalence of candida infections in critically ill non-immunocompromised patients has increased. This study aims to evaluate the utility of the use of serological tests (double immunodiffusion and Cand-Tec Test) for the determination of candidemia. METHODS: A retrospective evaluation is made of 214 patients admitted to the Intensive Care Unit (ICU) of the Agostino Gemelli University Polyclinic during a period of 42 months. The double immunodiffusion technique was utilized for the determination of Candida antibodies. The Cand-Tec latex agglutination test was performed to evaluate the presence of Candida antigen. Four hundred and fifty-five antigen and antibody tests were performed. RESULTS: Thirty-six patients (16.8%) developed an invasive candidiasis. The sensitivity and specificity of antibody detection tests was 29 and 67 respectively; the positive predictive value was 15 and the negative predictive value was 82. The sensitivity and specificity of the antigen detection test ranged between 82 and 3 and between 8 and 98 respectively according to different cut-off titre; the positive predictive value was low (13-17%) and the negative predictive value decreased from 70 to 29. CONCLUSIONS: The utility of the use of serological tests in the diagnosis of candidemia is extremely limited. The gold standard for the determination of Candida sepsis remains the demonstration of candida in blood or in tissues.


Asunto(s)
Candidiasis/diagnóstico , Femenino , Humanos , Masculino , Valor Predictivo de las Pruebas , Estudios Retrospectivos , Sensibilidad y Especificidad , Pruebas Serológicas
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