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"Background: Toll-like receptor is a pattern recognition receptor (PRR) that recognize pathogen-associated molecular pattern (PAMP) in a microorganism. Macrophages recognize the presence of mycobacteria through Toll-Like Receptor 2 (TLR2) and signaling further lead to the production of cytokines, both proinflammatory TNF-α, IL-1β, IL-6, IL-12, IL-15, IL-18 and IFN-γ, as well as anti-inflammatory IL4, IL-10 and TGF-β. TLR2 gene polymorphism is strongly determined by ethnicity and geography. Therefore it is necessary to uncovered the existence and association between Arg753Gln and Arg677Trp TLR2 gene polymorphism with TB susceptibility and its underlying mechanisms in Indonesian population in Bandung West Java. Methods: analytical observational study with cross-sectional design was conducted in Hasan Sadikin General Hospital Bandung from April 2011 to May 2012. Study population consisted of active pulmonary TB patient with positive AFB smear and Latent TB to ascertain previous MTb exposure. Polymorphism of gen Arg753Gln and Arg677Trp gene was determined with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. Plasma levels of IFN-γ, TNF-α, IL-10 and IL-12 were also compared between active and latent TB group. Results: heterozygote Arg753Gln TLR2 gene polymorphism was found in 9 of 86 pulmonary TB subjects (10.5%) but none in the latent TB group. The Arg677Trp polymorphism was not found in both groups. The odds ratio for Arg753Gln existence was 28.07 (p=0.022). No differences in the levels of IFN-γ, TNF-α, IL-10 and IL-12 between active pulmonary TB and latent TB subjects with and without Arg753Gln TLR2 gene polymorphism. Conlusion: Arg753Gln polymorphism of TLR2 gene is a risk factor for active pulmonary TB while Arg677Trp polymorphism is not. The Increased risk is not mediated by the difference in IFN-γ, TNF-α, IL-10 and IL-12 serum levels."

"Pemeriksaan dahak secara mikroskopik dengan pewarnaan Basil Tahan Asam (BTA) merupakan pemeriksaan yang sederhana, cepat, murah, dan cukup sensitif untuk mendukung diagnosis penyakit tuberkulosis serta untuk menilai kemajuan pengobatan. Penelitian ini bertujuan untuk menentukan metoda pewarnaan BTA terbaik yang dapat digunakan secara rutin terutama di laboratorium dengan beban pekerjaan yang cukup tinggi. Sensitivitas, spesifisitas, nilai prediksi positif dan negatif tiga macam metode pewarnaan BTA, yaitu Tan Thiam Hok, Ziehl Neelsen, dan Fluorokrom, dibandingkan terhadap hasil biakan dahak pada medium padat Lowenstein Jensen sebagai baku emas. Intepretasi hasil pewarnaan mengacu pada skala IUTLD. Pertumbuhan Mycobacterium tuberculosis didapatkan pada 27 dari 98 spesimen sputum (27,6%) berasal dari 98 penderita tersangka tuberculosis. Sensitivitas metoda pewarnaan Tan Thiam Hok, Ziehl Neelsen, dan Fluorokrom adalah 62,9%, 81,5%, dan 92,6%, sedangkan spesifisitasnya berturut-turut adalah 92,9%, 91,6%, dan 91,1%. Nilai prediksi positif berturut-turut adalah 77,3%, 78,6%, dan 71,4%, sedangkan nilai prediksi negatif adalah 86,8%, 92,9%, dan 96,8%. Dari penelitian ini didapatkan bahwa Ziehl Neelsen merupaka metoda terbaik dan dapat dilakukan di laboratorium sederhana.

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Comparison of Tan Thiam Hok, Ziehl Neelsen and Fluorochrome as Acid-Fast Bacilli Staining Methods in Sputum. Because of its simplicity, rapidity, low cost and relatively sensitive, sputum acid-fast bacilli (AFB) smear microscopy is the primary tool for detecting pulmonary tuberculosis and follow up of therapy. This experiment is aimed to determine the best acid-fast staining method that can be used for routine laboratory examination, especially in the high burdened clinical laboratory. We compared the sensitivity, specivity, and the positive and negative predictive value of 3 kinds of methods : Tan Thiam Hok, Ziehl Neelsen and Fluorochrome, using culture on Lowenstein-Jensen media as the gold standard. The smear results were observed using IUTLD scale. Twenty seven of 98 sputum specimens from 98 patients with clinical suspicion of tuberculosis (27,6 %) were positive by culture. The sensitivity of Tan Thiam Hok, Ziehl Neelsen and Fluorochrome were 62,9%, 81,5% and 92,6%, while the specivity were 92,9%, 91,6% and 91,1% respectively. The positive predictive value were 77,3 %, 78,6 %, 71,4 % , and the negative predictive value were 86,8 %, 92,9 %, 96,8 % respectively. Although fluorochrome gave the highest sensitivity, it needs special expensive equipments. We conclude that Ziehl Neelsen is still the method of choice for detecting AFB in sputum microscopically."

"ABSTRAKBackground: diagnostic of pulmonary TB in HIV patients is a problem due to non specific clinical features, or radiological appearance. HIV patients with CD4≤200 cells/mL infected with M. tuberculosis have less capacity in containing M. tuberculosis, developing granulomas, casseous necrosis, or cavities. This condition is caused by weakend inflammatory which later reduced sputum production and may cause false negative result. This study aimed to assess differences in the positivity level of acid fast bacilli (AFB) and cultures of M. tuberculosis from non-bronchoscopic sputum (spontaneous and induced sputum) compared to bronchoscopic sputum (bronchoalveolar lavage) in HIV positive patients suspected pulmonary tuberculosis with CD4<200 cells/μL.Methods: this cross sectional study was conducted in adult HIV patients treated in Hasan Sadikin Hospital with CD4≤200 cells/μL suspected with pulmonary tuberculosis by using paired comparative analytic test. All patients expelled sputum spontaneously or with sputum induction on the first day. On the next day, bronchoalveolar lavage (BAL) was performed. The two samples obtained from two methods were examined by AFB examination with staining Ziehl Neelsen (ZN) and cultured of M. tuberculosis on solid media Ogawa on all patients. Positivity, sensitivity and increased sensitivity of AFB and culture of M. tuberculosis in the non bronchoscopic and bronchoscopic groups were compared.Results: there were differences in the positivity level of AFB with ZN staining between non-bronchoscopic and bronchoscopic groups which were 7/40 (17.5%) vs 20/40 (50.0%) (p<0.001). The differences between the cultures of non-bronchoscopic and bronchoscopic groups were 16/40 (40.0%) vs 23/40 (57.5%) (p=0.039). Bronchoscopic sputum increased the positivity level of the ZN AFB examination by 32.5% (from 17.5% to 50.0%) as well as on culture examination by 17.5% (from 40.0% to 57.5%).Conclusion: Bronchoalveolar lavage can improve the positivity level of smears and cultures in patients suspected of pulmonary TB in HIV patients with CD4<200 cells/μL."

"ABSTRAKAdanya kerusakan sel hati pada infeksi virus Dengue (DENV) telah diketahuitetapi patogenesis yang mendasari hingga kini belum seluruhnya jelas. Penelitianini bertujuan untuk mengetahui lebih jelas patogenesis mekanisme yangmendasari kerusakan sel hati pada infeksi DENV dengan melihat efek langsungvirulensi virus dan efek tidak langsung.Kelompok penelitian terdiri dari: A. kontrol sel Huh7, B. Huh7 + DENV-2, C.Huh7 + antibodi NS-1, D. Huh7 + komplemen, E. Huh7 + antibodi NS-1 +komplemen, F. Huh7 + DENV-2 + antibodi NS-1 + komplemen, G. Huh7 +DENV-2 + antibodi NS-1, H. Huh7 + peripheral blood mononuclear cells(PBMC) dari pasien yang pernah terinfeksi DENV, I. Huh7 + DENV-2 + PBMCdari pasien yang pernah terinfeksi DENV, dan J. Huh7 + TNF-α. Aktivitas ASTdan ALT dari supernatan diukur sebagai indikator kerusakan sel Huh7.Median aktivitas pada sel Huh7 yang diberi DENV-2 0,1 moi selama 2 x 24 jam(AST 7 U/L dan ALT 3 U/L) lebih rendah bermakna dibandingkan kontrol (AST11 U/L dan ALT 4 U/L), yang menunjukkan efek anti-apoptotik DENV-2 padadosis 0,1 moi dan lama inkubasi 2 x 24 jam. Pada percobaan dengan berbagaidosis DENV dan lama inkubasi, didapatkan efek anti-apoptotik ini menghilangsetelah inkubasi 3 x 24 jam. Hasil tersebut menunjukkan kebenaran hipotesis efeklangsung merusak sel hati oleh infeksi DENV. Aktivitas AST 19 U/L dan ALT 8U/L pada sel Huh7 yang diberi antibodi NS-1 bersama komplemen lebih tinggibermakna dibandingkan kontrol sel Huh7 tanpa perlakuan (AST 11 U/L dan ALT4 U/L), maupun pemberian antibodi NS-1 saja (AST 10 U/L dan ALT 4 U/L).Hasil itu menunjukkan bahwa hipotesis molekular mimikri yang dibawakan olehantibodi NS-1 terbukti. Aktivitas AST 6,5 U/L pada pemberian antibodi NS-1bersama komplemen pada sel hati Huh7 yang sudah diinfeksi DENV-2 lebihrendah bermakna dibandingkan AST 11 U/L pada kontrol. Hasil itu menunjukkanDENV-2 memiliki efek anti-apoptotik yang mampu menghambat efek lisis dariantibodi bersama komplemen. Tidak terjadi peningkatan AST pada sel Huh7 yangdiberi PBMC saja (8 U/L), DENV-2 bersama PBMC (6 U/L) dan TNF-α saja (10U/L) dibandingkan kontrol (11 U/L).Dengan demikian, mekanisme kerusakan sel hati pada infeksi DENV in vitro yangdapat dibuktikan dengan penelitian ini adalah akibat efek langsung DENV-2 danmolekular mimikri pada sel hati yang dibawakan oleh antibodi NS-1.

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ABSTRACTThe involvement of liver cell damage in Dengue virus (DENV) infection isalready known but the whole pathogenesis is still not clear. Virus virulence isinvolved but other mechanisms may also play a role. This study aimed tounderstand the pathogenesis of liver cell damage caused by DENV infection,through direct pathogenic effect of the virus and indirect effect of immunologicalreaction.The study groups were A. Huh7 liver cells, control, B. Huh7 + Dengue virus-2New Guinea C strains (DENV-2), C. Huh7 + NS-1 antibody, D. Huh7 +complement, E. Huh7 + NS-1 antibody + complement, F. Huh7 + DENV-2 +NS-1 antibody + complement, G. Huh7 + DENV-2 + NS-1 antibody, H. Huh7 +peripheral blood mononuclear cells (PBMC) from previously DENV-infectedpatient, I. Huh7 + DENV-2 + PBMC from previously DENV-infected patient and,J. Huh7 + TNF-α. Activities of AST and ALT in the supernatant were measuredas indicator of liver cell damage.In the Huh7 cells, infected with DENV-2 0,1 moi (multiplicity of infection) andincubated for 2 x 24 hours, median AST 7 U/L and ALT 3 U/L were significantlylower compared to AST 11 U/L and ALT 4 U/L in Huh7 control cells. It wasconcluded that DENV-2 had anti-apoptotic effect at 0,1 moi dan incubation time 2x 24 hours. At various dosage of DENV-2 and incubation time study, the antiapoptoticeffect disappeared after 3 x 24 hours incubation; which means the effectwas temporary. The result proved the direct damaging effect of DENV to infectedliver cells. Activity of AST 19 U/L and ALT 8 U/L in Huh7 cells treated withNS-1 antibody together with complement, were significantly higher compared tocontrol (AST 11 U/L, ALT 4 U/L), and NS-1 antibody only (AST 10 U/L, ALT 4U/L). This result proved the molecular mimicry hypothesis by NS-1 antibody.Activity of AST 6,5 U/L in DENV-infected Huh7 cells treated with NS-1antibody and complement was significantly lower than AST 11 U/L in controlcells. This result indicated that DENV has anti-apoptotic effect that can inhibit thelytic effect of antibody and complement. There was no AST increase in Huh7treated with PBMC only (8 U/L), DENV-2 with PBMC (6 U/L) and TNF-α only(10 U/L) compared to control (11 U/L).The liver cell damage mechanisms of in vitro DENV infections which can beproven in this study are direct effect by DENV and molecular mimicry by NS-1antibody."