Functional variation of HIV-1 Rev Response Element in a longitudinally studied cohort

We showed previously that HIV-1 Rev Response Element (RRE) contains a certain degree of structural variation, and in a set of limited samples, RRE from HIV-1 natural isolates were found to have functional variability. The significance of the RRE heterogeneity is addressed further by analyzing the functional variation of RREs in a longitudinal cohort. While the RRE activity at early time points was not a good predictor of disease outcome, the RRE activity at late time points was correlated with rates of CD4+ count decline. These data suggest that RRE heterogeneity may be important in viral pathogenesis and disease progression.     

Chikungunya in Southeast Asia: understanding the emergence and finding solutions

In the last few years, chikungunya has become a major problem in Southeast Asia, with large numbers of cases being reported in Singapore, Malaysia, and Thailand. Much of the current epidemic of chikungunya in Southeast Asia is being driven by the emergence of a strain of chikungunya virus that originated in Africa and spread to islands in the Indian Ocean, as well as to India and Sri Lanka, and then onwards to Southeast Asia. There is currently no specific treatment for chikungunya and no vaccine is available for this disease. This review seeks to provide a short update on the reemergence of chikungunya in Southeast Asia and the prospects for control of this disease.     

Item Analysis to Improve Reliability for an Internal Medicine Undergraduate OSCE

Utilization of objective structured clinical examinations (OSCEs) for final assessment of medical students in Internal Medicine requires a representative sample of OSCE stations. The reliability and generalizability of OSCE scores provides validity evidence for OSCE scores and supports its contribution to the final clinical grade of medical students. The objective of this study was to perform item analysis using OSCE stations as the unit of analysis and evaluate the extent to which OSCE score reliability can be improved using item analysis data. OSCE scores from eight cohorts of fourth-year medical students (n = 435) in a 6-year undergraduate program were analyzed. Generalizability (G) coefficients of OSCE scores were computed for each cohort. Item analysis was performed by considering each OSCE station as an item and computing the corrected item-total correlation. OSCE stations which negatively impacted the reliability were deleted and the G-coefficient was recalculated. The G-coefficients of OSCE scores from the eight cohorts ranged from 0.48 to 0.80 (median 0.62). The median number of OSCE stations that negatively impacted the G-coefficient was 3.5 (out of a median of 25 total stations). When the ‘‘problem stations’’ were deleted, the median G-coefficient across eight cohorts increased to 0.62--0.72. In conclusion, item analysis of OSCE stations is useful and should be performed to improve the reliability of total OSCE scores. Problem stations can then be identified and improved.     

A child with avian influenza A (H5N1) infection

Human infections with avian influenza viruses can be severe and may be harbingers of the evolution of a pandemic strain. We present a patient in Thailand who was infected with influenza A (H5N1) virus. Prominent features included the progression from fever and dyspnea to the acute respiratory distress syndrome in a short period, lymphopenia and thrombocytopenia. Establishing the diagnosis for this patient increased public awareness of the virus and was soon followed by a halting of poultry-to-human transmission. On the basis of available data, any child with suspected avian influenza infection should be treated with oseltamivir.     

Immunophenotypic profile of adult acute myeloid leukemia (AML): analysis of 267 cases in Thailand

Little data exists in Thailand and other Southeast Asian countries regarding the biological characteristics of adult acute myeloid leukemia (AML). In this study, we performed a flow cytometric analysis of 267 Thai adult AML cases to delineate the pattern of leukemic cell surface antigens. Forty-eight cases (18%) were identified as acute promyelocytic leukemia (M3) and 219 cases as non-M3. The most frequent subtype of AML in Thailand was M1/M2 and the least frequent was M7. M3 immunophenotypes were characterized by their unique lack of expression of CD34 and HLA-DR as contrast to the high mean expression of 50% and 70%, respectively, in non-M3. Overall, 60% of cases expressed CD34. Aberrant lymphoid antigens were uniquely seen in specific subtypes of Thai AML, including CD19 (33% of non-M3 vs 23% of M3) and CD2 (12% of M3 vs 2% of non-M3). CD56 was frequently expressed in both M3 and non-M3 while CD16 appeared to be associated with M4/M5 (24% of cases) and CD7 with M1/M2 (21% of cases). Eighty-one percent of non-M3 expressed CD38 while only 53% of M3 did. We found that most Thai adult AML patients were on average 15-20 years younger than those of the West or Japan with only 25% of Thai cases over 60 years of age, although the immunophenotypes were not markedly different. Biological studies of acute leukemia in various countries should help to provide epidemiological clues that play a role in the pathogenesis of leukemia in different geographic regions of the world. Our study represents the largest series of AML ever investigated in the Southeast Asian region.     

In vivo tissue-specific regulation of the human papillomavirus type 18 early promoter by estrogen, progesterone, and their antagonists

Human papillomavirus type 18 is a causative agent of epithelial cancers in the uterine cervix. We show here that estrogen and progesterone activate beta-galactosidase expression from the early promoter of this virus in the genital epithelia of transgenic mice. Ovariectomy caused suppression of transgene expression exclusively in vagina and cervix epithelia. Beta-galactosidase expression could be restored in ovariectomized females by administration of estrogen, alone or in combination with progesterone. Further, rescue of transgene expression was inhibited by the estrogen antagonist tamoxifen and the anti-progesterone RU486, suggesting that this was a specific effect.     

Application of HIV-1-green fluorescent protein (GFP) reporter viruses in neutralizing antibody assays.

We made reporter HIV-1 DNA constructs carrying green fluorescent protein (GFP) gene and exchangeable env of subtype E. The recombinant constructs were used to produce infectious reporter viruses, which induced infected cells to emit green fluorescent light and rendered them easily detectable at single cell level. Because the env in this construct can be easily exchanged, viruses with different antigenic epitopes can be made. We used these reporter viruses to set up a neutralizing antibody assay based on fluorescence reduction by flow cytometric measurement. The result of this new assay correlated with the standard infectivity reduction assay using primary isolates. Because this new assay is faster and much less costly than the standard assay using a p24 endpoint and can be performed in peripheral blood mononuclear cells (PBMC), it provides a useful tool for analysis of HIV-1 immune responses.