More precise and rapid diagnostic methods for American cutaneous leishmaniasis (ACL) are necessary because of the growing number of cases observed in Brazil, including the northeastern region of the State of São Paulo. We applied PCR to 54 skin or mucosal biopsies from patients with a clinical and/or laboratory diagnosis of ACL using primers 13A and 13B, with positive results being obtained for 82% of the samples. When the PCR results were compared to those of histopathological leishmania detection, PCR showed superior results with 81.5% sensitivity and 95% CI of 68.0-95.1%. The Montenegro skin test (MST) was positive in 88.7% of patients. Since MST cannot be used as a diagnostic tool in endemic areas, the present results strongly suggest the use of PCR for the etiological confirmation of ACL, with emphasis on the mucosal form. Correspondence

Staphylococcus aureus is one of the principal human pathogens that colonize healthy individuals in the community in general, and it is responsible for severe infections in hospitalized patients. Due to an increase in the prevalence of strains of methicillin-resistant S. aureus (MRSA), combating these microorganisms has become increasingly difficult. A descriptive study was carried out on 231 patients in intensive care at the Oswaldo Cruz University Hospital (HUOC) in Recife, Brazil between January and April 2003 to determine the prevalence of S. aureus and MRSA and to evaluate risk factors for colonization by these bacteria when introduced into Intensive Care Units (ICUs). Body secretions were collected from the nostrils, axillary and perineal regions, and from broken skin lesions, of all patients during the first 48 hours following admission to the ICU. Samples were inoculated into blood agar and mannitol-salt-agar culture medium and identified by Gram staining, and by coagulase, DNAse and agglutination (Slidex Staph Test®) tests. Growth in Mueller-Hinton agar with 4% sodium chloride and 6mg/L oxacillin was used to identify MRSA. In addition, the latex agglutination test was performed to identify penicillin-binding protein, PBP 2A. The prevalence of S. aureus and MRSA was 87/231 (37.7%) and 30/231 (12.98%), respectively. There was no association between any risk factor studied (age, sex, origin of the patient -whether hospital or community, previous hospitalization, use of current or previous antibiotic therapy, corticotherapy and/or immunotherapy, reason for hospitalization and place of hospitalization) and the presence of S. aureus. However, a significant association was established between previous hospitalization and the presence of MRSA (RR:1.85; CI:1.00-3.41; p=0.041). The nostrils were the principal site of colonization by both S. aureus (80.4%) and MRSA (26.4%), followed by the perineal area, with rates of 27.6% and 12.6%, respectively. If only the nostrils had been investigated, the study would have failed to diagnose 17 patients (19.5%) as carriers of the pathogen into the ICU, thus contributing towards cross-dissemination.

In this study, we have the objective of evaluating the lymphoproliferative response and determining interferon (IFN)‐γ and interleukin (IL)‐10 cytokine production in the peripheral blood mononuclear cells (PBMC) of patients with American tegumentary leishmaniasis prior and post 12 months of chemotherapy treatment with meglumine antimoniate compared with the PBMC of noninfected donors. Lymphoproliferation, such as cytokine production, was evaluated through in vitro stimulus with the soluble antigenic fraction from Leishmania (Viannia) braziliensis promastigotes (1.25 µg/ml) and Concanavalin A (2.5 µg/ml). Patients showed a significant lymphoproliferative response prior and post treatment compared with the control group. Similar result, prior to chemotherapy treatment, was observed in IFN‐γ and IL‐10 production when patients were compared with the control group. After chemotherapy treatment, PBMC lymphoproliferative response of the patients revealed an increase, whereas patients have shown a decrease in IFN‐γ levels and an increase in IL‐10, although without statistical difference. These results may indicate that the patients produced a specific cellular response to the soluble antigenic fraction suggesting that besides Th1 and Th2 dichotomy, immunological regulation mechanisms with the participation of memory T cells and regulatory T cells could be present in the clinical evolution of these patients. This understanding will allow the study and identification of new L. (V.) braziliensis molecules potentially candidates to vaccines. J. Clin. Lab. Anal. 23:63–69, 2009. © 2009 Wiley‐Liss, Inc.

Studies suggest the influence of immune response on the successful treatment of American tegumentary leishmaniasis (ATL), and indicate the existence of protective immunity in self-healed patients. Thus, the aim of this work was to quantify interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), interleukin (IL-) 10, IL-17, IL-22 and nitric oxide (NO) in culture supernatants of PBMC from patients with active disease (AD), after treatment (AT), and from self-healed (SH) and healthy subjects (CT), in response to Leishmania (Viannia) braziliensis insoluble antigen (AgIns). All groups of patients produced IFN-γ, indicating a predominant proinflammatory profile. AD and AT patients presented TNF-α levels, with a slight increase after therapy, whereas it was weakly quantified in SH. Interestingly, NO secretion was significant in these individuals, whereas IL-17 appeared in low levels and seems to be regulated by NO. Although IL-22 was detected in AD, its role is still questionable. The presence of IL-10 in all groups of patients suggests that the cytokine plays distinct roles in the disease. These results indicate that specific cellular immunity takes part against Leishmania, but with some similarities between the different clinical states herein described; these mediators seem to be necessary for the cure to occur.