Dual candidemia detected by nested polymerase chain reaction in two critically ill children.

The use of improved microbiological procedures associated with molecular techniques has increased the identification of Candida bloodstream infections, even if the isolation of more than one species by culture methods remains uncommon. We report the cases of two children presenting with severe gastrointestinal disorders and other risk factors that contribute to Candida infections. In the first patient, C. albicans DNA was initially detected by a nested-amplification and C. tropicalis was found later during hospitalization, while blood cultures were persistently negative. In the second child, there was amplification of C. albicans and C. glabrata DNA in the same samples, but blood cultures yielded only C. albicans. Both patients received antifungal therapy but had unfavorable outcomes. These two cases illustrate that PCR was more successful than culture methods in detecting Candida in the bloodstream of high risk children, and was also able to detect the presence of more than one species in the same patient that might impact therapy when the fungi are resistant to azole compounds.

Preoperative embolization in mediastinal Castleman's disease.

Castleman's disease is a rare lymphoproliferative disease and its etiology is still unknown. It may occur at every site where lymph tissue is present. A definitive treatment is possible with complete resection. The most important problem is bleeding which may occur during surgery due to the high vascularization. In this study, we present the surgical treatment of a case with mediastinal Castleman's disease, treated preoperatively with embolization because of hypervascularization detected on thoracic CT.

Testicular Sertoli cell function in male systemic lupus erythematosus.

OBJECTIVE: To assess the testicular Sertoli cell function in male SLE patients. METHODS: Thirty-four consecutive patients were prospectively selected to evaluate serum inhibin B. Clinical features, treatment, semen analysis, urological evaluation, testicular ultrasound, hormones and anti-sperm antibodies were determined. RESULTS: Patients were subdivided into two groups: low serum inhibin B (Group 1, n = 8) and normal levels (Group 2, n = 26). The median sperm concentration (P = 0.024), total sperm count (P = 0.023) and total motile sperm count (P = 0.025) were lower in Group 1. Inhibin B levels were positively correlated with sperm concentration (r = 0.343), total motile sperm count (r = 0.357), and negatively correlated with follicule-stimulating hormone (FSH) (r = 0.699) and luteinizing hormone (r = 0.397). The median serum inhibin B was lower in SLE patients treated with intravenous cyclophosphamide (IVCYC) compared with those without this therapy (P = 0.031). Further evaluation of the 26 SLE patients with normal inhibin B and FSH levels revealed that medians of inhibin B/FSH ratio were lower in SLE patients with oligozoospermia compared with normozoospermia (P = 0.004). This ratio was also lower in SLE patients treated with IVCYC than those without this therapy (P = 0.04). In contrast, inhibin B serum level alone did not discriminate the later group of patients (P = 0.12). CONCLUSIONS: This is the first study to identify a high frequency of testicular Sertoli cell dysfunction in male SLE associated with semen abnormalities. Further prospective studies are necessary to determine if inhibin levels and inhibin B/FSH ratio will be an earlier and useful marker of IVCYC toxicity in these patients.

Suitability of a rapid DNA isolation and amplification for detection of Trypanosoma cruzi in Triatoma infestans dry fecal spots collected on filter paper.

A rapid DNA extraction was used for T. cruzi detection in triatomines dry fecal spots collected on filter paper and analyzed by PCR. Fifty T. infestans were fed on experimentally infected Balb/C mice with high T. cruzi parasitemia and divided into five groups of ten triatomines, and 100 triatomines were infected with lower parasitemia and divided into five groups of 20 triatomines. One dry fecal spot was analyzed per group on days 1, 2, 3, 4 and 5 post feeding. Amplification targeted T. cruzi TCZ sequence and resulted positive from day 4 after bugs feeding in the two models (high and lower parasitemial. The rapid DNA isolation and PCR proposed are suitable for detection of T. cruzi DNA in filter paper and should be considered in field research.

Contribution of mitral valve reserve capacity to sustained symptomatic improvement after balloon valvulotomy in mitral stenosis: implications for restenosis.

OBJECTIVES: To explain the discrepancy between the symptomatic status of patients and the hemodynamically calculated mitral valve area during long-term follow-up after mitral balloon valvulotomy, mitral valve orifice variability after dobutamine infusion was investigated in two groups of patients. BACKGROUND: A significant increase in aortic valve area with increased aortic transvalvular flow has been reported in patients with calcific aortic stenosis after aortic balloon valvulotomy. A similar phenomenon with regard to the mitral valve has not been studied in detail. METHODS: Group 1 comprised 10 patients (mean age 33 +/- 9 years) with untreated mitral stenosis. Group 2 comprised 29 consecutive patients (mean age 32 +/- 7 years) who underwent successful percutaneous mitral balloon valvulotomy 13 +/- 2 months before the study. RESULTS: After dobutamine infusion, heart rate and cardiac index increased significantly in both groups. The mean pulmonary artery pressure, mitral valve gradient and pulmonary capillary pressure remained unchanged in Group 2 but increased significantly in Group 1. The mean mitral valve area was significantly larger in Group 2 after dobutamine infusion than at baseline (1.9 +/- 0.5 vs. 2.4 +/- 0.6 cm2, p < 0.0001) but was unchanged in Group 1 (1.2 +/- 0.2 vs. 1.3 +/- 0.3 cm2, p = NS). The mean mitral valve area in seven patients in Group 2 (24%) was < or = 1.5 cm2 before dobutamine infusion (1.3 +/- 0.4 cm2), which was defined as restenosis. In five of these seven patients who had minimal or no symptoms, the mitral valve area increased significantly after dobutamine infusion (1.3 +/- 0.1 vs. 1.9 +/- 0.1 cm2). In the other two patients who were symptomatic, the mitral valve area did not change after dobutamine infusion. These two patients were identified as having "true" restenosis, and redilation of the mitral valve was performed in both. CONCLUSIONS: In patients who underwent mitral balloon valvulotomy, increased mitral valve reserve capacity contributed to symptomatic improvement on long-term follow-up. Dobutamine infusion may be helpful in detecting clinically significant restenosis.

Myasthenia gravis: how to treat?

Myasthenia Gravis is an acquired autoimmune disorder caused by a neuromuscular transmission defect which is clinically characterized by fluctuating weakness of voluntary muscles and fatigability. It can be diagnosed by clinical features, clinical, pharmacological and electrophysiological tests and serological evaluation. Treatment modalities include symptomatic treatment in the form of cholinesterase inhibitors and plasmapheresis and immunotherapy in the form of immunosuppressant medications, immunomodulating therapy and thymectomy. No single regimen is appropriate for all patients and up to now no mode of therapy has been proven to be clearly superior. The response to any form of treatment is difficult to assess because the severity of symptoms fluctuate. We retrospectively analyzed the clinical records of 33 myasthenia gravis patients which were managed at our clinic between 1995-2003. All patients were treated with anticholinesterase medications sometime during their treatment. Most patients recieved immunosupressant and/or immunomodulator therapy. Patients were referred for thymectomy when indicated. We evaluated the outcome with different treatment modalities, focusing on the role of thymectomy. We also investigated the possible correlations between clinicopathological features and clinical outcome. We conclude that as for the medical treatment of myasthenia gravis azathioprine plus steroid improves the outcome; and for the surgical treatment, early thymectomy should be performed in all generalize myasthenia patients.

A case of cardiac hydatid cyst localized on the interventricular septum and causing pulmonary emboli.

Cardiac hydatid cyst is rarely encountered. In this article, a case of hydatid cyst localized in multiple organs including the ventricular septum and causing pulmonary emboli is reported.

Serum tumor necrosis factor levels in acute myocardial infarction and unstable angina pectoris.

Tumor necrosis factor (TNF) enhances leukocyte adherence to vascular endothelium and increases procoagulant activity in the endothelial cells. Thus it may be implicated in the pathogenesis of acute vascular occlusions. To study the role of TNF in the early stages of acute myocardial infarction (MI), the authors measured circulating TNF levels in the sera of patients with acute MI and unstable angina pectoris. Blood samples were obtained within six hours after onset of chest pain and stored at -70 degrees until tested. A sensitive sandwich enzyme-linked immunosorbent assay (ELISA) test was used for TNF measurement. C-reactive protein (CRP) levels were determined semiquantitatively. Immediate complications such as heart failure, arrhythmia, and shock were also noted. Twenty-four patients with electrocardiographically and biochemically confirmed acute MI and 14 patients with unstable angina pectoris were included in the study. TNF levels were serially assessed at the time of admission and at hours 6, 24, 48, 72, and 96 after onset of chest pain in 2 patients with acute MI. Detectable TNF was found in 13 sera of the acute MI group (range; 10-1510 pg/mL) and 4 sera of the angina pectoris group (range; 15-240 pg/mL). There was no correlation between the serum TNF levels and the occurrence of complications and the extent of myocardial damage. CRP response was unrelated to TNF levels. Contrary to previous reports, serial measurement of TNF revealed that peak values were reached within six hours and disappeared after twenty-four hours.(ABSTRACT TRUNCATED AT 250 WORDS)

A case of arrhythmogenic right ventricular dysplasia associated with FANA positiveness--a case report.

This is the case report of a forty-five-year-old woman who manifested clinical findings characteristic of arrhythmogenic right ventricular dysplasia (ARVD) except for fluorescent antinuclear antibody (FANA) positiveness. To the authors' knowledge, this is the first report of FANA positiveness associated with ARVD.

PCR in the first oropharynx aspirate of the newborn: a possible source for identification of congenital infection agents.

We present a case of prenatal diagnosis of congenital rubella. After birth, in addition to traditional serologic and clinical examinations to confirm the infection, we could identify the virus in the "first fluid aspirated from the oropharynx of the newborn", using polimerase chain reaction (PCR). We propose that this first oropharynx fluid (collected routinely immediately after birth) could be used as a source for identification of various congenital infection agents, which may not always be easily identified by current methods.

Experimental infection and horizontal transmission of Bartonella henselae in domestic cats.

In order to study B. henselae transmission among cats, five young cats were kept in confinement for two years, one of them being inoculated by SC route with B. henselae (10(5) UFC). Only occasional contact among cats occurred but the presence of fleas was observed in all animals throughout the period. Blood culture for isolation of bacteria, PCR-HSP and FTSZ (gender specific), and BH-PCR (species-specific), as well as indirect immunofluorescence method for anti-B. henselae antibodies were performed to confirm the infection of the inoculated cat as well as the other naive cats. Considering the inoculated animal, B. henselae was first isolated by blood culture two months after inoculation, bacteremia last for four months, the specific antibodies being detected by IFI during the entire period. All contacting animals presented with bacteremia 6 months after experimental inoculation but IFI did not detect seroconversion in these animals. All the isolates from these cats were characterized as Bartonella (HSP and FTSZ-PCR), henselae (BH-PCR). However, DNA of B. henselae could not be amplified directly from peripheral blood by the PCR protocols used. Isolation of bacteria by blood culture was the most efficient method to diagnose infection compared to PCR or IFI. The role of fleas in the epidemiology of B. henselae infection in cats is discussed.

Tc-99m sestamibi gated SPECT in patients with left bundle branch block.

PURPOSE: The purpose of this study was to assess the diagnostic role of a Tc-99m sestamibi gated SPECT technique in patients with left bundle branch block (LBBB) without known coronary artery disease. MATERIALS AND METHODS: Twenty consecutive patients with constant complete LBBB were included. A same-day rest-stress protocol was used, and dipyridamole stress (14 patients) or treadmill exercise (6 patients) was applied. Electrocardiograph (ECG)-gated SPECT images were acquired 15 minutes after the administration of 0.31 mCi/kg Tc-99m sestamibi at peak stress. Regional myocardial perfusion was analyzed in relation to the cardiac cycle. RESULTS: Eleven of 14 patients who underwent a dipyridamole stress test had hypoactivity in the left anterior descending (LAD) artery territory in the ungated (summed) stress-rest images (abnormality ratio, 78%). On the ungated images, the abnormality was completely reversible in one patient (9%), partially reversible in five patients (46%), irreversible in two patients (18%), and reverse perfusion was identified in three patients (27%). Abnormality ratios of end-systolic and end-diastolic data were 93% and 29%, respectively. Conversely, the ungated rest-stress and end-systolic images of all the patients who performed treadmill exercise were abnormal despite the presence of normal or nearly normal end-diastolic myocardial perfusion. The angiographic findings correlated best with those of end-diastolic images. In 13 patients without coronary artery disease, normal or nearly normal regional perfusion was observed on end-diastole, but four patients with abnormal end-diastolic perfusion, which involved the LAD territory in all but one, had substantial coronary artery disease. The number of the involved segments was similar on the end-systolic and ungated data. Most of these artifactual defects were localized to the anteroseptal, septal, and inferoseptal segments. CONCLUSIONS: These preliminary data indicate that end-diastolic images can significantly reduce artifactual defects in patients with LBBB. The resolution of an LBBB pattern on end-diastolic data would significantly improve the diagnostic role of myocardial perfusion studies in these patients.

[Inter- and intra-cephalic variations in pathogenicity in Toxoplasma gondii. Clinical and epidemiologic consequences]. / Variations inter et intra-cépales de pathogénicité chez Toxoplasma gondii. Conséquences cliniques et épidémiologiques.

Clinical and epidemiological studies, especially the ones conducted in HIV+ patients indicate that the pathogenicity of Toxoplasma gondii varies according to the strain considered. The differences observed among strains are represented by distinct genomic DNA patterns which could be experimentally evaluated by means of the RFLP (Restriction Fragment Length Polymorphism) and the RAPD (Random Amplified Polymorphic DNA). If on the one hand virulent strains present very similar genomic DNA patterns, on the other hand chronic strains are highly polymorphic. These differences may be, at least in part, due to the asexual (clonal) multiplication of virulent strains. The existence of an intraspecific pathogenicity variation and genetic heterogeneity was observed within a single strain either after attenuation (infection in Fischer or Wistar rats), or during reactivation (in immunodepressed animals). In a congenital model of toxoplasmosis, the differences are detected from one animal to another and sometimes, even from one organ to another in the same host. This finding do not seem to be related to the occurrence of mutations but rather to selective pressures, notably of immunological origin, exerted by the infected organism. A better understanding of these phenomena could result in significant therapeutic and prophylactic advances. Our first effort will be directed to the establishment of more precise diagnostic and predictive elements. The accomplishment of this step relies on the use of primers deriving from DNA sequences characteristic of virulence and which will be tested by PCR.

Comparison of two commercial immunoassays for the detection of anti-rubella IgM and IgG in pediatric patients.

The only threatening modality of rubella is the Congenital Rubella Syndrome that affects fetuses of women who acquire infection during early pregnancy. Laboratory diagnosis is based on serological parameters. We compared anti-rubella IgM and IgG detection of two commercial immunoassay kits (Abbott and Roche). Although we observed an agreement of 97.8% for IgM and 95.7% for IgG when the categories positive, negative and indeterminate were considered, mean titers of IgG and the absorbance/cut off of IgM were statistically different for both kits, thus corroborating the idea that serological results depend very much on the methodology and must be carefully interpreted.

Collection tubes with or without gel separator did not interfere with detection of rubella virus antibodies IgM and IgG.

Rubella infection is an exanthematic disease, with high prevalence in the adult population. The only modality of disease that causes serious consequences is congenital rubella syndrome (CRS), which happens when a pregnant woman seronegative to rubella virus acquires the infection during early pregnancy. Due to the lack of signals and characteristic symptoms of disease, diagnosis of rubella is based essentially on laboratory tests: antibodies detection and/or virus isolation. Results of serologic tests should always be interpreted with caution, because they can be affected by the quality of blood samples, processing and storage of sera, the equipment and reagents used to perform tests, and finally by the technical expertise and training of biologists. The collection tubes with gel seem to facilitate serum separation, but on the other hand gels can retain and consequently decrease antibody titers. Therefore, we decided to investigate whether the use of collection tubes containing gel separator might interfere with rubella virus antibody detection in blood samples from children. We did not observe statistically significant differences with respect to rubella virus antibody detection (immunoglobulin M [IgM] and immunoglobulin G [IgG]) for samples collected in tubes with or without gel separator, from the two evaluated manufacturers.