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ABSTRACT: This study describes the genotype distribution of Pneumocystis jiroveci in 79 respiratory samples obtained from 15 patients with acquired immunodeficiency syndrome (AIDS) with P. jiroveci pneumonia and 64 human immunodeficiency virus-negative subjects with different chronic pulmonary diseases. The genotyping was based in analysis of 2 independent genetic loci: the mitochondrial large subunit ribosomal RNA (mt LSU rRNA) fragment (assessed by direct sequencing) and the gene for dihydropteroate synthase (DHPS; assessed by restriction fragment-length polymorphism). The mt LSU rRNA analysis revealed the presence of 3 different polymorphisms for both populations. The major genotype, 85C/248C, was found to be significantly higher in patients with AIDS and P. jiroveci pneumonia than in patients with pulmonary disease. The rate of genotypes 85A/248C and 85T/248C was similar in both groups. The analysis of DHPS genotypes assesses the prevalence of its 4 possible genotypes, with 35.5% of genotypes related to sulfa resistance. The data suggest a common source of infection between both groups.
Clinical Infectious Diseases 08/2004; 39(1):123-8. · 9.15 Impact Factor
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ABSTRACT: A total of 251 clinical specimens (235 gastric aspirates and 16 bronchoalveolar lavages) from 88 children were prospectively tested in a blinded manner for the presence of Mycobacterium tuberculosis complex, by use of the Amplicor M. tuberculosis test and by means of in-house polymerase chain reaction (PCR). The results were compared with those obtained by conventional culture and by direct microscopy. All of the children underwent extended follow-up to verify or exclude the clinical diagnosis of tuberculosis. The results of the different tests, when compared to the final clinical diagnosis, were a sensitivity of 60% and a specificity of 96.8% for in-house PCR, 44% and 93.7% respectively for the Amplicor test, 44% and 100% for mycobacterial culture and 12% and 100% for microscopy. Amplicor tests presented false-positive findings in children without tuberculous infection. We conclude that both in-house PCR and the Amplicor test are rapid methods that can be helpful for difficult or urgent diagnosis of tuberculosis in children. However, efforts should be aimed toward improvement of the sensitivity and specificity of an easy-to-use PCR kit.
Clinical Infectious Diseases 02/2001; 32(1):17-22. · 9.15 Impact Factor
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ABSTRACT: The production of free radicals seems to be involved in the mechanisms of ototoxicity. Aminoglycosides produce ototoxicity, which can be determined through distortion product otoacoustic emissions (OAEs) that measure the activity of the outer hair cells of the organ of Corti. An ototoxic chart was obtained in rats using gentamicin or tobramycin. Together with this treatment, the animals ingested melatonin in the drinking water, or melatonin was injected subcutaneously or intramuscularly. The distortion product OAEs were determined over a prolonged period of time for each of the groups. The effect of melatonin on the antibiotic capacity of the aminoglycosides used was also studied. Antibiograms inoculated with Escherichia coli or Pseudomonas aeruginosa and treated with gentamicin or tobramycin in the presence or absence of melatonin at quantities from pharmacological to physiological doses were performed. The ototoxicity produced by gentamicin and tobramycin was maximal from days 3 to 5 post-treatment, returning to normal values in 2 wk. When melatonin was present, the recovery was at day 5 post-treatment, independently of the means of administration of the pineal product. The antibiograms showed that melatonin had no effect on the antibiotic capacity. It is concluded that the ototoxicity caused by gentamicin and tobramycin is ameliorated by melatonin and that the pineal hormone does not interfere with the antibiotic capacity of these antibiotics.
Journal of Pineal Research 02/2000; 28(1):26-33. · 5.79 Impact Factor
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ABSTRACT: We investigated the value of the polymerase chain reaction (PCR) in the diagnosis of active tuberculosis in children and evaluated the relationship between PCR results in children with tuberculous infections and mediastinal adenopathies detected by computerized tomography (CT-Scan). This was a controlled, blinded, prospective study comparing nested PCR, mycobacterial cultures and the clinical diagnosis based on 350 clinical specimens from 117 children referred for evaluation of suspected pulmonary tuberculosis. All children with tuberculous infection but without active disease underwent a chest CT-scan to detect the presence of mediastinal adenopathies not evident on chest x-ray. The sensitivity of PCR was 56.8% in children with clinically active disease (culture: 37.8%; smears: 13.5%). A major advantage of PCR over cultures was noted when there was no parenchymal involvement on chest radiograph and when the patient was undergoing anti-tuberculous treatment. There were nine specimens with false-negative PCR results due to the presence of amplification reaction inhibitors. PCR was positive in five children with tuberculous infection without active disease and these children presented mediastinal adenopathies on the CT-scan that were not evident on chest radiography. There were no false-positive PCR results in the control groups of children. We conclude that nested PCR is a rapid and sensitive method for the early diagnosis of tuberculosis in children. It is especially useful when the diagnosis of active tuberculosis is difficult. In our study children with tuberculous infection without apparent disease who have positive PCR results have mediastinal adenopathies on CT-scan.
Pediatric Pulmonology 12/1999; 28(5):344-51. · 2.53 Impact Factor
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ABSTRACT: Fever of intermediate duration (FID), characterized by a febrile syndrome lasting from 7 to 28 days, is a frequent condition in clinical practice, but its epidemiological and etiologic features are not well described. Murine typhus (MT) is a worldwide illness; nevertheless, to our knowledge, no studies describing its epidemiological and clinical characteristics have been performed in the south of Spain. Also, its significance as a cause of FID is unknown.
To determine the epidemiological features, clinical characteristics, and prognosis of MT and, prospectively, its incidence as a cause of FID.
Prospective study of cases of MT over 17 years (1979-1995) and of all cases of FID treated in a tertiary teaching hospital in Seville, Spain.
One hundred and four cases of MT were included, and MT was the cause in 6.7% of 926 cases of FID. Insect bites were reported in only 3.8% of the cases of MT previous to the onset of illness. Most cases (62.5%) occurred in the summer and fall. A high frequency of rash (62.5%) was noted. Arthromyalgia (77%), headache (71%), and respiratory (25%) and gastrointestinal (23%) symptoms were also frequent. Laboratory findings were unspecific. Organ complications were uncommon (8.6%), but they were severe in 4 cases. The mean duration of fever was 12.5 days. Cure was achieved in all cases, although only 44 patients received specific treatment.
Murine typhus is prevalent in the south of Spain and is a significant cause of FID. Clinical signs are benign, but some patients may develop severe complications. A high degree of clinical suspicion is required for diagnosis.
Archives of Internal Medicine 05/1999; 159(8):872-6. · 11.46 Impact Factor
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ABSTRACT: A unusually high rate of HCV-infected individuals in whom the HCV genotype cannot be ascertained by means of single PCR and LIPA procedures has recently been reported in our area. The aim of the present study was to investigate the epidemiological, clinical and molecular characteristics of these patients.
Cross-sectional study. Eighty anti-HCV-positive patients with chronic liver disease, 45 (56.25%) of them intravenous drug users, were included. HCV genotyping was carried out in all patients using commercial single PCR and LIPA procedures. Samples where no HCV RNA amplification and/or indeterminate HCV genotype were found were also tested by means of a nested PCR. HCV viral load was measured in all patients.
HCV genotyping was not achieved in 23 (28.75%) individuals. No amplification of HCV RNA was found in 19 of them, and in four other cases the LIPA procedure did not allow identification of a distinct HCV genotype. After the use of nested PCR+LIPA, it was found that the HCV genotype 4 was found in 11 of those 23 individuals (47.82%). Ten of these 11 HCV genotype 4-harboring individuals were intravenous drug users. The HCV viral load was lower in HCV genotype 4-harboring individuals than in those whom the genotypes 1, 2 or 3 were found (p<0.001).
A high rate of HCV genotype 4-harboring cases has been found among HCV-infected individuals in Southern Spain. Had only single PCR been used, these individuals could have been wrongly regarded as non-viremic.
Journal of Hepatology 07/1997; 27(1):25-9. · 9.26 Impact Factor
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B Soto,
A Sánchez-Quijano,
L Rodrigo,
J A del Olmo,
M García-Bengoechea,
J Hernández-Quero,
C Rey,
M A Abad,
M Rodríguez,
M Sales Gilabert,
F González,
P Mirón,
A Caruz,
F Relimpio, R Torronteras,
M Leal,
E Lissen
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ABSTRACT: To investigate the possible role of HIV infection in the natural history of chronic parenterally-acquired hepatitis C.
A multicenter cross-sectional study was performed in 547 patients with chronic parenterally-acquired hepatitis C with or without HIV infection (116 HIV-positive and 431 HIV-negative). Approximate duration of HCV infection was estimated in all patients included, and histologic diagnoses made at different time intervals following HCV infection were analyzed in both groups. Factors related to serum HCV-RNA levels were also investigated.
Histologic findings were similar in liver biopsies from both HIV-infected and noninfected patients. However, in the first 10 years, 13 out of 87 (14.9%) HIV-positive subjects developed cirrhosis, in comparison with 7 out of 272 (2.6%) in the HIV-negative group (p < 0.01). Similar results were found in the first 5 and 15 years, respectively, and most of the HIV-negative patients with cirrhosis (42 out of 56) developed cirrhosis in a time interval longer than 15 years. Consequently, mean interval from estimated time of HCV infection to cirrhosis was significantly longer in HIV-negative than HIV-positive patients (23.2 vs. 6.9 years; p < 0.001). Chronic active hepatitis (with and without cirrhosis) and long duration of HCV infection were significantly associated with higher HCV load (p < 0.05). Finally, HIV-positive patients with CD4+ cell counts > 500 cells/ml showed a lower HCV load than those with < 500 cells/ml (p < 0.05).
HIV infection modifies the natural history of chronic parenterally-acquired hepatitis C with an unusually rapid progression to cirrhosis. HIV-related immunodeficiency may be a determinant of higher hepatitis C viremia levels and more severe liver damage.
Journal of Hepatology 02/1997; 26(1):1-5. · 9.26 Impact Factor
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ABSTRACT: Two hundred and ninety-four non-transfused prostitutes from Spain, who denied intravenous drug abuse, were tested for hepatitis C virus (HCV) antibodies. Seventeen (5.78%) of them were seropositive. Both in univariate and correspondence analyses, serological results for hepatitis C were associated with the HIV-1 serostatus (p < 0.001), number of sex partners (p < 0.05) and a history of genital ulcers (p < 0.05). In logistic regression analysis, hepatitis C seropositivity was associated only with HIV-1 infection (adjusted odds ratio = 13.6; 95% confidence interval = 3.3-55.2). These results show that hepatitis C seropositivity is associated with HIV-1 infection in female non-intravenous drug abusing prostitutes. These findings are also consistent with the hypothesis that HCV can be sexually transmitted with low efficiency.
European Journal of Clinical Microbiology 05/1995; 14(5):460-4. · 2.86 Impact Factor
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J. A. Pineda,
A. Rivero,
C. Rey,
J. Hernández-Quero,
A. Vergara,
J. Muñoz,
I. Aguado,
J. Santos, R. Torronteras,
J. A. Gallardo,
C. García-Arias
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ABSTRACT: Two hundred and ninety-four non-transfused prostitutes from Spain, who denied intravenous drug abuse, were tested for hepatitis C virus (HCV) antibodies. Seventeen (5.78 %) of them were seropositive. Both in univariate and correspondence analyses, serological results for hepatitis C were associated with the HIV-1 serostatus (p
European Journal of Clinical Microbiology 04/1995; 14(5):460-464. · 2.86 Impact Factor
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The Lancet 02/1995; 345(8942):127. · 38.28 Impact Factor
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ABSTRACT: To investigate the concordance of anti-hepatitis C virus (anti-HCV) reactivity by a second-generation enzyme immunoassay (EIA-2) and by a four-antigen recombinant immunoblot assay (4-RIBA) in homosexual men, in comparison with that found in other sexually active groups and blood donors.
Prospective study.
Tertiary referral centre, Seville, Spain.
A total of 1203 subjects were studied. Eight hundred and three were sexually active individuals: 547 female prostitutes, 88 heterosexual men who had frequent sexual intercourse with prostitutes, and 168 homosexual men. All of them denied blood transfusion and parenteral drug use. In addition, 400 voluntary blood donors were selected at random.
All serum samples were screened for anti-HCV by EIA-2 and repeatedly reactive sera were tested by 4-RIBA. Homosexual men were also screened for anti-human immunodeficiency virus (anti-HIV), hepatitis B virus (HBV) markers and gammaglobulin concentration. Finally, serum samples from homosexual men reactive for anti-HCV by EIA-2 were analyzed for HCV-RNA by polymerase chain reaction (PCR).
Concordance between EIA-2 and 4-RIBA in female prostitutes (71.4%), clients of prostitutes (75.0%), and blood donors (83.3%) was significantly higher than in homosexual men (38.8%) (P < 0.04). In this collective the concordance between 4-RIBA and PCR was 85.7% for positive cases and 88.8% for negative ones, and EIA-2 ratios in reactive sera were significantly higher in 4-RIBA confirmed cases (P < 0.0001). No correlation between false positive EIA-2 results and presence of HIV infection, HBV markers or hypergammaglobulinaemia was found in homosexual men by univariate analysis.
There is a high level of non-specific anti-HCV reactivity by EIA-2 amongst homosexual men in comparison with that found in other sexually active groups and blood donors. The true prevalence of HCV infection amongst homosexual men could be even lower than previously described.
Journal of Internal Medicine 07/1994; 236(1):73-7. · 5.48 Impact Factor
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ABSTRACT: To determine the predictive value of IgM anti-hepatitis C virus (HCV) testing in patients with chronic hepatitis C infections undergoing interferon-alpha (IFN-alpha) therapy, IgM anti-HCV reactivity was analysed by two different methods (non-commercial and commercial) in 19 patients and monitored at times 0 (pretreatment), 3, 6, 12, and 24 months during follow-up. Eight patients were non-responders, five remained in sustained response 1 year after stopping treatment, and six had a relapse. No correlation between alanine transaminase (ALT) levels and IgM anti-HCV reactivity was found by either method in baseline samples. In addition, neither the presence nor absence of IgM anti-HCV in baseline samples, nor the loss of specific IgM reactivity during treatment, had any predictive value. Finally, no other parameters analysed (age, sex, risk group and histological diagnosis), were significantly associated with IgM anti-HCV reactivity in our study. In summary, these results suggest that baseline detection and monitoring of IgM anti-HCV reactivity are not useful in predicting the sustained response to IFN-alpha therapy in chronic hepatitis C infection.
Journal of Viral Hepatitis 02/1994; 1(2):159-61. · 4.09 Impact Factor
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ABSTRACT: To define the role of sexual transmission in the spread of hepatitis C virus (HCV) infection, a seroprevalence study of antibodies against HCV was performed in populations at high risk for sexually transmitted diseases. Subjects included 310 female prostitutes, 88 clients of prostitutes, 168 homosexual men and 147 stable heterosexual partners of index cases reactive for anti-HCV (98 of whom were partners of drug addicts coinfected with HCV and human immunodeficiency virus [HIV]). All subjects denied prior transfusion or intravenous drug use. Controls were 400 voluntary blood donors selected randomly from first-time donors. The prevalence of anti-HCV by enzyme immunoassay, confirmed by a second-generation recombinant immunoblot assay, was 6.4% in prostitutes, 6.8% in clients of prostitutes, 4.2% in homosexual men, 7.4% in heterosexual partners of index cases and 1.2% in random donors. However, the anti-HCV prevalence in stable heterosexual partners of HCV-positive/HIV-positive index cases was 2.2 times higher than in stable heterosexual partners of index cases reactive for anti-HCV only (9.2% vs. 4.1%), and sexual partners of index cases coinfected with HCV and HIV were almost three times more likely to be infected with HIV than with HCV (25.5% vs. 9.2%). These data suggest that HCV infection may be sexually transmitted but with low efficiency and that this efficiency could be increased in the presence of coexistent HIV infection in the index case.
European Journal of Clinical Microbiology 12/1993; 12(11):827-31. · 2.86 Impact Factor
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ABSTRACT: Endocarditis by Q fever is a diagnostic and therapeutic challenge given the diagnostic delay and elevated morbidity and mortality it carries. Six cases of endocarditis by Q fever attended over the last 7 years were retrospectively studied. Five patients had been previously diagnosed of valvular involvement and three had prosthesis. Five patients presented a febrile syndrome of prolonged duration with negative hemocultures and progressive valvular changes. One patient presented acute valvular failure requiring emergency surgery. The most significant laboratory data were anemia, thrombocytopenia, high ESR and hypergammaglobulinemia. In the echocardiograms valvular vegetations were observed in 4 cases. All the patients received medical treatment with doxicylin, one associated with rifampicin and another cotrimoxazol. In 4 patients valvular reposition was required due to a severe hemodynamic alteration. After a minimum follow up of 2 years all the patients remain asymptomatic. The serologic evolution is described.
Medicina Clínica 06/1993; 100(17):664-7. · 1.38 Impact Factor
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ABSTRACT: The finding of an important proportion of cases of hepatitis C without previous contact with blood or hemoderivates has led to suspect that there may be other routes of transmission among which sexual transmission may be found.
The presence of antibodies against the hepatitis C virus (anti-HCV) and the association of this infection with certain epidemiologic parameters and sexual practice was determined in 184 homosexual males with no other risk factors for virasis of intravenous transmission. Moreover, the prevalence of anti-HCV was evaluated in 210 voluntary blood donors. Every homosexual was surveyed on sexual practice and the first serum sample available of each of these patients was analyzed for anti-HVC and anti-HIV-1.
Twenty-two homosexual (12%) were anti-HCV positive while only one (0.5%) of the control was positive for this marker (p < 0.0001). Thirty-six cases (19%) were anti-HIV positive (none of the controls). No statistical association was found between both serologic markers or between the presence of anti-HCV and the epidemiologic parameters evaluated.
Homosexuals constitute a risk group for hepatitis C virus infection although the sexual route is probably not effective for transmission of this entity.
Medicina Clínica 01/1993; 100(2):50-2. · 1.38 Impact Factor
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Transactions of the Royal Society of Tropical Medicine and Hygiene 90(4):383. · 2.16 Impact Factor
