A paired comparison of tuberculin skin test results in children with clinical manifestations of tuberculosis using 2 TU and 5 TU tuberculins

Rationale: A paired comparison of reactivity to purified protein derivative 2 TU PPD RT-23 and 5 TU PPD-S in children with clinical manifestations of tuberculosis was done to assess if 2 TU PPD RT-23 can be used instead of 5 TU PPD-S in routine Mantoux testing in the country. Objective: To determine the correlation of skin test reactivity to 2 TU PPD RT-23 and 5 TU PPD-S. Study Design: Cross Sectional Study. Methods: Two simultaneous skin tests using 2 TU PPD RT-23 and 5 TU PPD-S were performed. Each dose was randomly assigned in a blinded manner to the right or left forearm and read after 72 hours. Correlation between the size of induration obtained with 2 TU PPD RT-23 and with 5 TU PPD-S was done, as well as, correlation between tuberculin reactivity and age, gender, nutritional status, presence of BCG vaccination, exposure, and clinical manifestations. A p-value <0.05 was considered statistically significant. Results: Sixty five patients were included in the study. The overall mean difference in paired reaction sizes for the two doses was-1.02 + 2.8 mm (range of -11 to 3 mm). Using the present guidelines in the country to determine a positive tuberculin skin test, 27 (41.5 %) patients were positive when tested with 2 TU PPD RT-23 and 33 (50.8 %) patients were positive when tested with 5 TU PPD. The mean PPD size with 2 TU was 4.7 mm + 6.1 mm compared to 5.8 mm + 6.1 mm with 5 TU. PPD skin test reactivity with the two reagents was highly correlated (intraclass correlation 0.88; 95% CI 0.83-0.94). There was no significant association between age, gender, nutritional status, presence of BCG vaccination, TB exposure, and clinical manifestations to tuberculin reactivity. Conclusion: Tuberculin skin test reactivity among children, who were with clinical manifestations of tuberculosis and tested with 2 TU PPD RT-23 and 5 TU PPD-S, were found to be comparable. Age, gender, nutritional status, presence of BCG vaccination, TB exposure, and clinical manifestations were not factors influencing the size of the PPD reaction. 2 TU PPD RT-23 can be used instead of 5 TU PPD-S in routine Mantoux testing.

An in-vitro study on the antibacterial effect of neem (Azadirachta indica) leaf extract on methicillin-sensitive and methicillin-resistant Staphylococcus aureus

The most common cause of pyogenic infection of the skin and soft tissues in children is Staphylococcus aureus, a fast-emerging problem due to its accompanying significant cost and morbidity. The popularity of herbal medications has increased due to the search for cheaper and more accessible alternatives. However, data is still lacking to back up these claims. Although a few in vitro studies have tested Neem leaf extract on S. aureus, there are none done on Methicillin-resistant Staphylococcus aureus (MRSA) despite the fact that it is being marketed for such purposes. Objectives: This study aims to determine if Neem leaf extract (Azadirachta indica) has antibacterial properties against Methicillin-sensitive and Methicillin-resistant Staphylococcus aureus and to compare the anti-staphylococcal properties of Neem leaf extract with oxacillin, vancomycin, mupirocin, and povidone iodine. Methods: An in vitro experimental study was performed using Neem leaf, properly identified and verified, was subjected to ethanol extraction of its active ingredients then diluted to produce 25%, 50%, 75%, and 100% concentrations. Standard strains of Staphylococcus aureus and clinical isolates of MRSA where inoculated on blood agar plates and subjected to the standardized disc susceptibility testing method. Zones of inhibition were measured for each test extract and compared to currently used medications, namely oxacillin, vancomycin, mupirocin, and povidone iodine with the pure diluent as negative control. The data was analyzed using difference of means hypothesis testing; it utilized the student's t-test to determine significance. Results: A trend of increasing antibacterial activity was noted with increasing concentration of the extract. Zones of inhibition started to appear at 50% concentration for S. aureus and 75% for MRSA. The antibiotics were able to produce greater zones of inhibition than the Neem extracts. Conclusion: Data from this study strongly suggest that the ethanol extract from Neem leaves exhibits in vitro antibacterial activity against both Staphylococcus aureus and MRSA with greatest zones of inhibition noted at 100% concentration.

N-acetyltransferase2 genotype and its association with hepatitis among Filipino children treated with first line anti-tuberculosis drugs

RATIONALE: Among the first line antituberculosis (anti-TB) drugs, the major drug incriminated in the development of hepatotoxicity is isoniazid (INH). The human N-acetyl transferase2 (NAT2) gene is mainly responsible for INH metabolism. This gene exhibits a hereditarily determined polymorphism. There is presently no study on the predominant NAT2 genotype among Filipinos. There are also no Filipino studies on the incidence of hepatitis and other adverse effects of first line anti-TB drugs. OBJECTIVES: To determine the predominant NAT2 genotype and its association with the development of hepatitis among Filipino children given first line anti-TB drugs (INH, rifampicin and pyrazinamide) and to determine the incidence of hepatitis and other serious adverse reactions to these drugs. STUDY DESIGN: Prospective cohort study SETTING: Tertiary government hospital in Metro Manila STUDY POPULATION: Children on to 18 years old with pulmonary tuberculosis and normal liver function test at baseline. METHODS: Total bilirubin (TB), direct bilirubin (DB) and liver transaminases (AST and ALT) were checked routinely at baseline and at thow, four, eight and 12 weeks after starting treatment. Within the first month of treatment, blood was also taken for NAT2 genotyping. The identification of the three NAT2 polymorphisms that are associated with a slow acetylator status - 481C to T (NAT2*5), 950G to A (NAT2*6) and 857G to A (NAT2*7) was carried out by polymerase chain reaction-restriction fragment length polymorphism. All patients were followed up for a total of six months. The presense of any adverse effects like gastroinstestinal symptoms, rash, hepatitis or drug fever was also monitored. RESULTS: A total of 24 children [mean age: 5 years; 11 males] were included. Majority (96%) were diagnosed by passive detection and mean Z score was - 1.38 (1 to -3). No patient developed hepatotoxicity or any side effects to anti-TB drugs. In 23 patients who had NAT2 genotyping, 39% and 22% were alleles homozygous for the NAT2*6 and NAT2*7, respectively. There was a combination of alleles in only three (13%) subjects. CONCLUSION: NAT2*6 and NAT2*7 alleles associated with a slow acetylator status were detected among our patients although the presence of these variants did not lead to any hepatotoxicity nor any treatment-related side effects. A larger study with broader genotype analysis is needed to confirm the present findings.