Our results suggest that the QFT-GIT assay may be a sensitive tool for the detection and prediction of active tuberculosis in HIV-1-infected individuals.

To investigate the presence of diarrheagenic Escherichia coli in Lambaréné , Gabon, 150 children with diarrhea were screened for enteroaggregative E. coli (EAEC), enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), and enteroinvasive E. coli (EIEC) using polymerase chain reaction and an HEp-2 cell culture techniques. Isolates of EAEC were detected in 57 children, two thirds of them between six months and two years of age, and isolates of ETEC were detected in seven patients. Isolates of EPEC, EHEC, and EIEC were not present in this population. Among the EAEC, the pCVD432 plasmid, a heat stable (ST)−like (ST-like) enterotoxin (EAST), and a plasmid-encoded heat-labile toxin (PET) were detected in 19, 34, and 42 cases, respectively. Detection of pCVD432, EAST, and PET were significantly associated with EAEC identified by the HEp-2 cell assay. Although detected only in 16 patients, the presence of the fimbriae AAF I (aagA) and AAF II (aafA) were more likely to occur in EAEC than in non-EAEC (odds ratio [OR] ‫ס‬ 4.1, 95% confidence interval [CI] ‫ס‬ 0.5−38.6, and OR ‫ס‬ 2.3, 95% CI ‫ס‬ 1.0−5.3, respectively). The EAEC isolates exhibited decreased susceptibility for ampicillin, tetracycline, and trimethoprim.

We report on an HIV-negative but immunocompromised patient with disseminated acanthamoebiasis, granulomatous, amoebic encephalitis, and underlying miliary tuberculosis and tuberculous meningitis. The patient responded favorably to treatment with miltefosine, an alkylphosphocholine. The patient remained well with no signs of infection 2 years after treatment cessation.A 25-year-old man from India, who had been living in Austria for 7 years and had no previous history of major illnesses, was brought by ambulance to the hospital for dyspnea, cough, fever, and weight loss. During neurologic examination, a hearing impairment was suspected. The patient was unable to walk because of severe ataxia. Skin examination showed several necrotic ulcers with purulent discharge and black eschars, measuring 0.5 cm to 3 cm, located on the skull, back, neck, and arms ( Figure 1, panels A and B). Miliary tuberculosis (TB) of the lungs, liver, spleen, and kidneys was suspected on the basis of chest radiography and computed tomography (CT) of chest and abdomen. Ziehl-Nielsen (ZN) staining for acid-fast bacilli in sputum, bronchial secretions, and lavage obtained through bronchoscopy was negative. PCR for Mycobacterium tuberculosis in bronchial secretions and serum was positive. Culture on Loewenstein agar resulted in growth of nonresistant M. tuberculosis after 31 days. Blood cultures were negative for aerobic/anaerobic bacteria, mycobacteria, and fungi. Results of serologic tests were negative for Aspergillus, Candida, Cryptococcus, Histoplasma, Blastomyces, and Coccidioides spp. Severe immunosuppression with a CD4+ lymphocyte count of 182 cells/μL made HIV infection probable, but HIV testing results were negative. Cranial CT showed multiple small enhancing lesions in cerebral cortex and underlying white matter, pons, midbrain, and around most of the cisterns. On magnetic resonance imaging (MRI), the lesions appeared as high T2 signal areas that enhanced heterogeneously or in a ringlike manner. These fi ndings were compatible with the diagnosis of meningoencephalitis with intracerebellar abscess formation.Cerebrospinal fl uid (CSF) obtained through lumbar puncture was negative for Toxoplasma gondii, Encephalitozoon cuniculi, and Enterocytozoon bieneusi by PCR and for Trypanosoma gambiense by indirect hemagglutination assay. Staining and antigen testing (enzyme immunoassay) for Cryptococcus neoformans was negative, as was Treponema pallidum antibody testing. No viruses (herpes simplex 1 and 2, varicella zoster, enterovirus) could be detected by PCR. Cultures were negative for aerobic/ anaerobic bacteria and fungi. ZN staining detected acid-fast bacilli that were confi rmed to be nonresistant M. tuberculosis after culture for 38 days. PCR for M. tuberculosis was positive. An Acanthamoeba-specifi c PCR (1) and DNA sequencing of the PCR product showed Acanthamoeba genotype T2 (corresponding to group III). High immunoglobulin (Ig) G (2,000) and IgM (1,000) titers against Acanthamoeba spp. could be demonstrated serologically. The organism ...

Acanthamoeba amoebae of genotype T2 were identified as the causative agent of Acanthamoeba skin lesions and granulomatous amoebic encephalitis (GAE) in a human immunodeficiency virus-negative patient with underlying tuberculosis. To our knowledge this, is the first case of GAE involving genotype T2.Acanthamoebae are the causative agents of Acanthamoeba keratitis and granulomatous amebic encephalitis (GAE), a condition that predominantly occurs in immunocompromised individuals. In GAE the skin and the olfactory neuroepithelium probably function as portals of entry, and occasionally, inflammation can be observed at these primary foci. Descriptions of approximately 150 cases of GAE caused by Acanthamoeba have been published worldwide, and less than 10 of these patients have survived (9). Recently, the first case of GAE in Austria was diagnosed (A. Aichelburg et al., submitted for publication). The patient was a 25-year-old man from India with respiratory and neurological symptoms and widespread granulomatous dermal lesions. He had a CD4 ϩ count of 182 cells/l but was negative for human immunodeficiency virus (HIV). The final diagnosis included miliary tuberculosis, tuberculous meningitis, GAE, and Acanthamoeba skin lesions. The patient did not improve after tuberculostatic treatment or treatment with streptomycin, fluconazole, trimethoprim-sulfamethoxazole, amphotericin B, flucytosine, and sulfadiazine, which are drugs that are potentially effective against Acanthamoeba. However, he was finally treated successfully with oral and topical miltefosine and intrathecal and intravenous amikacin.Altogether six cerebrospinal fluid (CSF) samples, two dermal punch biopsy specimens, one bronchoalveolar lavage (BAL) specimen, one lung tissue biopsy specimen, one liver tissue biopsy specimen, and one biopsy specimen from the main brain lesion were investigated for acanthamoebae by lactophenol cotton blue (LPCB) staining (15), culture, and PCR. For culture, specimens were inoculated onto the centers of nonnutrient agar plates (1.5%) precoated with Escherichia coli. The plates were sealed with Parafilm, incubated at 30°C for 14 days, and examined every 48 h. For PCR, whole-cell DNA was isolated as described previously (19) and a fragment of the 18S rRNA gene was amplified with primers JDP1 and JDP2 (8). Acanthamoeba sp. strain ATCC PRA-105, genotype T4, was used as a positive control. The amplicons were sequenced with a 310 ABI Prism automated sequencer (Applied Biosystems, Langen, Germany), and the sequences of both strands were obtained. Multiple-sequence alignment was performed with the Clustal X application (16) but with the primer sites excluded from the analysis. Genotypes were assessed with the model assumption of a Ͻ5% sequence dissimilarity within one genotype (5). The lung, liver, and brain biopsy specimens were also examined by immunofluorescence staining with polyclonal rabbit anti-Acanthamoeba castellanii serum at dilutions of 1:50 and 1:100. Sequential patient serum samples at dilutions ranging from 1:5 to 1:4,000 w...

Flow cytometry for the intracellular detection of T-cell cytokines was performed for 15 Gabonese patients during acute uncomplicatedPlasmodium falciparum malaria. A striking expansion of CD4+ and CD8+ T cells producing gamma interferon (IFN-γ) was found during drug-induced clearance of parasitemia, paralleled by a decrease of interleukin-2 (IL-2) production. The frequency of IL-4- and IL-13-producing CD4+cells gradually decreased, whereas the frequency of T cells producing IL-2+–IFN-γ+, IL-4−–IL-5+, and IL-4+–IL-5+ cytokines as well as IL-4+–IFN-γ+ and IL-13+–IFN-γ+ cytokines was not significantly altered. The capacity for IL-10 production within the CD4+ subset increased due to an expansion of both IL-10+–IFN-γ− and IL-10+–IFN-γ+ cytokine-expressing cells. Thus, a more pronounced Th2-driven immune response during acute untreated P. falciparum infection with a shift towards Th1 responsiveness induced by parasite clearance is suggested.