SUMMARYHuman Immunodeficiency Vírus Type 1 and 2 antibodies detection was performed in 457 dried whole blood spots samples (S&S 903). Q-Preven HIV 1+2 was the screening test used. The results were compared with the gold standard serum tests by ELISA (Cobas Core e Axsym HIV1/2 gO) and imunofluorescence was the definitive confirmatory test.The samples were obtained from the Hospital Nossa Senhora da Conceição in Porto Alegre, RS -Brazil, through whole blood transfer to filter paper card and sent to Caxias do Sul, RS -Brazil where the tests were performed.The dried whole blood spot stability was evaluated with two different panels. The first one was composed of five negative and five positive samples stored at room temperature, 4 °C, -20 °C and -70 °C, while the second was composed of two negative and three positive samples stored at 37 °C (humidity <50%). Each sample was screened every week for six weeks. These measurement results didn't show variation during the study period.The detected sensibility was 100%, specificity was 99.6%, the positive predictive value was 99.5% and negative predictive values were 100%. The results demonstrated high performance characteristics, opening a new perspective of dried whole blood spot utilization in HIV screening diagnosis.

Endothelial dysfunction is common in T1DM adolescents with less than 5 yr of disease. It is associated with duration of disease, microalbuminuria, and mean second-year HbA1c but not with mean first-year HbA1c. These data support the metabolic memory hypothesis.

SUMMARYIn Brazil, the existing reference values for T-lymphocytes subsets are based on data originated in other countries. There is no local information on normal variation for these parameters in Brazilian adults and children. We evaluated the normal variation found in blood donors from five large Brazilian cities, in different regions, and in children living in Salvador, and Rio de Janeiro. All samples were processed by flow cytometry. The results were analyzed according to region, gender, and lifestyle of blood donors. A total of 641 adults (63% males), and 280 children (58% males) were involved in the study. The absolute CD3+, and CD4+ cells count were significantly higher for females (adults and children). Higher CD4+ cell count in adults was associated with smoking, while higher CD8+ count was found among female children. Higher counts, for all T-cells subsets, were detected in blood donors from southeast / south regions while those living in the northern region had the lowest values. Individuals from midwestern and northeastern regions had an intermediate count for all these cells subsets. However, these differences did not reach statistical significance. In Brazil, gender and smoking, were the main determinants of differences in T-lymphocytes reference values.

The utilization of cryopreserved peripheral blood mononuclear cells (PBMC) for immunologic assays has recently increased. These cells are particularly useful in clinical trials with low end-point frequencies, because they allow the performance of assays after the complete identification of the end points (1,8). Cryopreserved PBMC can be assayed in batches, avoiding inter-and intralaboratory variabilities when they represent potential confounders (6).The use of cryopreserved PBMC in immunological assays poses challenges, including the availability of adequate equipment and the need for technical proficiency. Assays must be adapted and validated for the use of cryopreserved PBMC, and the quality of the frozen cells has to be monitored to ensure reliable results in functional and phenotypic assays. We have previously shown that the results of functional assays are strictly dependent on the viability of the cryopreserved PBMC (9), such that Ͻ70% viability compromises lymphoproliferative responses to antigens and mitogens. PBMC with viability of Ն70% are also suitable for cytokine production studies, flow cytometric analyses, and immunomagnetic cell separation (4,5,7,9). While cell recovery does not interfere with the results of immunologic assays, the recovery of only a small proportion of cells may preclude assay performance altogether. Based on these observations, cryopreservation quality assurance (QA) programs must monitor the viability and recovery of cryopreserved PBMC. We report here on an effort to assess and improve the ability of seven Pediatric AIDS Clinical Trials Group (PACTG) sites in Brazil to cryopreserve viable PBMC.This program included seven Brazilian sites, the Division of AIDS Immunology QA (IQA) Program, and the PACTG Cryopreservation Working Group. Pertinent aspects of this study were approved by local institutional review boards and the Brazilian National Research Council. The PACTG cryopreservation consensus protocol (http://impaact.s-3.com /immlab.htm) was distributed to the sites and discussed in detail over conference calls. The first three QA rounds used PBMC from 5 to 10 healthy volunteers per site. Cryopreserved PBMC were stored in liquid nitrogen at the sites and shipped to the IQA on dry ice. The time elapsed during transportation and the amount of dry ice in the package upon arrival were recorded by the IQA. At the IQA, thawed PBMC were assessed for viability and recovery. Companion vials were also thawed at the sites to evaluate the effects of shipment on viability and recovery. Troubleshooting occurred with bimonthly conference calls. After the first three shipments, technologists from five sites participated in a wet-laboratory workshop at the IQA for hands-on training. After the workshop, QA rounds included cells from human immunodeficiency virus (HIV)-infected and uninfected volunteers. A second wet-laboratory workshop was organized 1 year after the first one.The viability of PBMC thawed according to the PACTG cryopreservation consensus protocol was measured manually by the try...

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In a four-year period (July/2001-June/2005), 410 Haemophilus spp. isolates were studied. Those were isolated from sputum at Hospital Nossa Senhora da Conceição (NSC) in Porto Alegre city (RS). β β β β β-lactamase enzyme was detected in 113 (27.6%) of isolates through chromogenic cephalosporin method. Fifty-eight (51.3%) of them showed sensibility to ampicillin through disc-diffusion method using Haemophilus Test Medium (HTM) by NCCLS criteria. In 297 (72.4%) isolates β β β β β-lactamase was not detected by chromogenic cephalosporin method. Five (1.7%) of them were resistant and 1 (0.3%) intermediate to ampicillin using disc-diffusion method. The authors emphasized the importance of Haemophilus spp. resistance to ampicillin research in clinical laboratories routine and the use of more than one method for this analysis was proposed, due to different resistance mechanisms in Haemophilus spp.