G6PD Deficiency at Sumba in Eastern Indonesia Is Prevalent, Diverse and Severe: Implications for Primaquine Therapy against Relapsing Vivax Malaria

Safe treatment of Plasmodium vivax requires diagnosis of both the infection and status of erythrocytic glucose-6-phosphate dehydrogenase (G6PD) activity because hypnozoitocidal therapy against relapse requires primaquine, which causes a mild to severe acute hemolytic anemia in G6PD deficient patients. Many national malaria control programs recommend primaquine therapy without G6PD screening but with monitoring due to a broad lack of G6PD deficiency screening capacity. The degree of risk in doing so hinges upon the level of residual G6PD activity among the variants present in any given area. We conducted studies on Sumba Island in eastern Indonesia in order to assess the potential threat posed by primaquine therapy without G6PD screening. We sampled 2,033 residents of three separate districts in western Sumba for quantitative G6PD activity and 104 (5.1%) were phenotypically deficient (\u3c4.6U/gHb; median normal 10U/gHb). The villages were in two distinct ecosystems, coastal and inland. A positive correlation occurred between the prevalence of malaria and G6PD deficiency: 5.9% coastal versus inland 0.2% for malaria (P\u3c0.001), and 6.7% and 3.1% for G6PD deficiency (P\u3c0.001) at coastal and inland sites, respectively. The dominant genotypes of G6PD deficiency were Vanua Lava, Viangchan, and Chatham, accounting for 98.5%of the 70 samples genotyped. Subjects expressing the dominant genotypes all had less than 10% of normal enzyme activities and were thus considered severe variants. Blind administration of anti-relapse primaquine therapy at Sumba would likely impose risk of serious harm

Antibodies In Sera Of Dengue Patients With Plasma Leakage Cross-Reacting With DENV Protein And Endothelial Protein

BACKGROUND: Dengue infection remains a major public health problem in Indonesia. Severe dengue associated with plasma leakage require hospitalization and potentially life threatening. However, the mechanism remains unclear, and the occurrence is unpredictable. The role of anti-endothelial antibody is predicted play an important role in the pathogenesis of plasma leakage, as severe dengue is more prevalent in secondary infection or post vaccinated individuals.METHODS: Serum samples from 127 single Dengue Virus (DENV) serotype infected subjects were obtained in day 2 of fever onset. Subjects were divided into plasma leakage and non-plasma leakage based on World Health Organization (WHO) criteria. Anti-endothelial antibody in patient sera were detected using western blot of Human Umbilical Vein Endothelial Cells (HUVEC). To confirm cross-reactivity, the sera was preabsorb with mix-DENV lysate. RESULTS: Three prominent bands were identified on western blot strips that inhibited by pre-absorption with DENV lysate. Plasma leakage patient expressed significantly more antibodies, with 51.7% of plasma leakage patients expressed at least two bands out of those three, compared to 18.5% of non-plasma leakage. CONCLUSION: Antibodies found in sera of dengue patients with plasma leakage cross-reacted with DENV proteins and endothelial proteins 37 kDa, 75 kDa, 120 kDa, and therefore may be involved in the pathogenesis of plasma leakage. Proteomic identification of those protein targets is needed and may be useful for vaccine studies and further development of predictor marker for plasma leakage in dengue

Antibodies in Sera of Dengue Patients with Plasma Leakage Cross-Reacting with DENV Protein and Endothelial Protein

BACKGROUND: Dengue infection remains a major public health problem in Indonesia. Severe dengue associated with plasma leakage require hospitalization and potentially life threatening. However, the mechanism remains unclear, and the occurrence is unpredictable. The role of anti-endothelial antibody is predicted play an important role in the pathogenesis of plasma leakage, as severe dengue is more prevalent in secondary infection or post vaccinated individuals.METHODS: Serum samples from 127 single Dengue Virus (DENV) serotype infected subjects were obtained in day 2 of fever onset. Subjects were divided into plasma leakage and non-plasma leakage based on World Health Organization (WHO) criteria. Anti-endothelial antibody in patient sera were detected using western blot of Human Umbilical Vein Endothelial Cells (HUVEC). To confirm cross-reactivity, the sera was preabsorb with mix-DENV lysate.RESULTS: Three prominent bands were identified on western blot strips that inhibited by pre-absorption with DENV lysate. Plasma leakage patient expressed significantly more antibodies, with 51.7% of plasma leakage patients expressed at least two bands out of those three, compared to 18.5% of non-plasma leakage.CONCLUSION: Antibodies found in sera of dengue patients with plasma leakage cross-reacted with DENV proteins and endothelial proteins 37 kDa, 75 kDa, 120 kDa, and therefore may be involved in the pathogenesis of plasma leakage. Proteomic identification of those protein targets is needed and may be useful for vaccine studies and further development of predictor marker for plasma leakage in dengue.KEYWORDS: severe dengue, plasma leakage, cross-reactive, anti-endothelial antibody

High Adenosine Deaminase Level and Erythrocyte Sedimentation Rate of Intestinal Tuberculosis Patients

BACKGROUND: Currently, laboratory diagnosis of intestinal tuberculosis (ITB) is limited based on clinical manifestations, providing opportunities for alternative laboratory tests to diagnose ITB. At the present time, the role of serum adenosine deaminase (ADA) and hematological tests in ITB patients are not widely known. The objective of this study was to determine the role of ADA and hematological tests in patients suspected with ITB.METHODS: Subjects that were suspected of ITB were classified as ITB group, while subjects with inflammatory bowel disease, hemorrhoid, and intestinal malignancy were classified as non-ITB group. Colonoscopy, histopathological examinations, and hematological test were performed. ADA measurement was also performed with clinical chemistry analyzer based on enzymatic colorimetry principle.RESULTS: Out of 143 subjects, 16 (11.2%) subjects were diagnosed with ITB and 127 (88.8%) subjects were classified as non-ITB group. ADA level and erythrocyte sedimentation rate (ESR) of ITB group were significantly higher than the ones of non-ITB group (p<0.05). Cut-off, sensitivity, and specificity of ADA level were 12.56 IU/L, 75%, and 57%, respectively. Cut-off, sensitivity, and specificity of ESR were 32.5 mm/hour, 81%, and 62%, respectively. Colonoscopy of ITB subjects displayed multiple ulcerations, edema, and hyperemic mucosa. Histopathological examination of ITB subjects exhibited granulomatous inflammation, epitheloid cells, giant cells, and lymphocyte aggregates.CONCLUSION: ADA level and ESR were significantly higher among ITB patients compared with non-ITB patients. Since the sensitivities of ADA and ESR tests were high, the ADA and ESR tests could be considered as a screening test for ITB.KEYWORDS: intestinal tuberculosis, adenosine deaminase, hematological test

Comparison of Light Transmission Aggregometry and VerifyNow in Detecting Clopidogrel Resistance and Factors Affecting Clopidogrel Resistance in AMI-EST Patients Undergoing Percutaneous Coronary Intervention: A Cross-Sectional Study

BACKGROUND: Light transmission aggregometry (LTA) and VerifyNow is commonly used to measure platelet responsiveness to clopidogrel. This study aimed to compare the results of LTA and VerifyNow P2Y12 assay for assessing the clopidogrel resistance in patients undergoing percutaneous coronary intervention and determine factors affecting clopidogrel resistance.METHODS: The subjects were 119 patients who underwent percutaneous coronary intervention (PCI) and had given loading dose of 600 mg clopidogrel. Blood samples were taken at 6 hour after clopidogrel loading dose. Platelet aggregation was measured by LTA and VerifyNow.RESULTS: LTA and VerifyNow assay showed fair agreement with Kappa=0.270, p=0.001. The proportion of resistance to clopidogrel using VerifyNow was 21.8% and LTA was 47.1%. Patients with diabetes melitus were more likely to develop clopidogrel resistance than patients without diabetes (OR of 7.67; 95% CI: 1.87-31.50; p=0.005).CONCLUSION: The ability of LTA and VerifyNow in detecting clopidogrel resistance were not comparable. Multivariate analysis results for VerifyNow shows diabetes mellitus as the greatest predictors of clopidogrel resistance.KEYWORDS: agreement, clopidogrel resistance, LTA, predictor, VerifyNow

Exceptional Complex Chromosomal Rearrangements in Three Generations

We report an exceptional complex chromosomal rearrangement (CCR) found in three individuals in a family that involves 4 chromosomes with 5 breakpoints. The CCR was ascertained in a phenotypically abnormal newborn with additional chromosomal material on the short arm of chromosome 4. Maternal karyotyping indicated that the mother carried an apparently balanced CCR involving chromosomes 4, 6, 11, and 18. Maternal transmission of the derivative chromosome 4 resulted in partial trisomy for chromosomes 6q and 18q and a partial monosomy of chromosome 4p in the proband. Further family studies found that the maternal grandmother carried the same apparently balanced CCR as the proband’s mother, which was confirmed using the whole chromosome painting (WCP) FISH. High resolution whole genome microarray analysis of DNA from the proband’s mother found no evidence for copy number imbalance in the vicinity of the CCR translocation breakpoints, or elsewhere in the genome, providing evidence that the mother’s and grandmother’s CCRs were balanced at a molecular level. This structural rearrangement can be categorized as an exceptional CCR due to its complexity and is a rare example of an exceptional CCR being transmitted in balanced and/or unbalanced form across three generations

Demographic, malaria and G6PDd prevalence data by gender and ecosystem in western Sumba.

<p>n = sample number.</p><p>¹ Population number was obtained from central agency statistic from each district.</p><p>² Microscopy diagnosed.</p><p>³ One subject experienced mixed infection of <i>P</i>. <i>falciparum</i> and <i>P</i>. <i>vivax</i>.</p><p>⁴ Hb < 10 g/dl.</p><p>⁵ G6PD activities ≤4.6 U/gHb.</p><p>⁶ Samples screened as G6PD deficient but declined, absent or failed to be DNA extracted.</p><p>Demographic, malaria and G6PDd prevalence data by gender and ecosystem in western Sumba.</p

A) Schematic work flow of the study in western Sumba in 2012 and B) map of G6PD deficiency and malaria prevalences in study sites in Sumba.

<p>Small inlet showed the map of Indonesia and where Sumba island is located.</p

Impact of Hb level on G6PD activities.

<p>Blue line represents those samples having extremely high G6PD activity and some degree of anemia (R² = 0.08) and red line represents those having normal Hb level (R² = 0.48) and t-value = 28.6 (p<0.001).</p