Hasil Pencarian  ::  Simpan CSV :: Kembali

"[ABSTRAKLatar Belakang. SNP IL-28B mempunyai peran penting dalam pencapaian SVR pada pengobatan Hepatitis C kronik antarras manusia dan berpotensi untuk memprediksi keberhasilan terapi Peg-IFN/RBV maupun penyembuhan spontan Hepatitis C akut. Hingga saat ini, mekanisme molekular yang mendasari kaitan SNP IL-28B dengan respons terapi masih belum jelas meskipun diperkirakan terkait dengan ekspresi IFN-l3 dan reseptor IFN-l3 di jaringan hati.Tujuan. Mengetahui hubungan SNP IL-28B dan SVR serta ekspresi IFN-l3 dan reseptor IFN-l3 di jaringan hati serta mendapatkan kemaknaan klinis SNP IL-28B dan kovariat SVR dalam memprediksi respons terapi Peg-IFN/RBV.Metode. Penelitian ini terbagi menjadi dua bagian. Pertama, penelitian potong lintang pada pasien Hepatitis C kronik yang telah selesai menjalani terapi Peg-IFN/RBV dengan melakukan pengambilan data dasar dan sampel darah. Kedua, penelitian kasus kontrol pasien yang menjalani biopsi hati dan pewarnaan imunohistokimia.Hasil. Pencapaian SVR yang lebih tinggi ditemukan pada pasien dengan alel CC SNP IL28B (p=0,014). Alel CC SNP IL28B mempunyai ekspresi IFN-l3 lebih tinggi dibandingkan dengan alel non-CC (p = 0,018). Meskipun demikian, tidak ditemukan adanya perbedaan bermakna antara ekspresi IFN-l3 (p = 0,237) maupun reseptor IFN-l3 dengan SVR (p = 0,237). Pada penelitian ini, diformulasikan persamaan faktor risiko pencapaian SVR sebagai p = 1 / (1 + e-y); e = 2,7, y = -2,498 + 2,652 (SNP IL-28B) + 2,029 (trombosit) untuk praterapi dan sedangkan untuk masa terapi y = -0,223 + 2,621 (RVR).Simpulan. SNP IL-28B merupakan faktor risiko praterapi yang penting dalam pengobatan hep C kronik G1 menggunakan terapi dua kombinasi. Alel mayor IL28B mengekspresikan IFN-l3 dan reseptornya lebih banyak sebagai respons adanya VHC, namun tidak ditemukan adanya hubungan hal tersebut dengan pencapaian SVR. RVR merupakan faktor masa terapi terbaik untuk memprediksi SVR. Penelitian lanjutan diperlukan untuk membuktikan adanya faktor lain yang berperan dalam pencapaian SVR.;

---

ABSTRACTBackground: SNP IL-28B played an important role in achieving sustained virological response (SVR) among different ethnics in chronic Hepatitis C patients and is considered potential in predicting treatment response of Pegylated interferon/ribavirin (Peg-IFN/RBV) combination and spontaneous clearance in acute hepatitis. Up to date, molecular mechanism underlying correlation between SNP IL28B and SVR has not been fully understood yet although it is predicted to be related to IFN-λ3 and IFN-λ3 receptor in liver tissues.Aim: Understanding the association between SNP IL-28B and SVR in chronic Hepatitis C treatment and expression of IFN-l3 and IFN-l3 receptor in liver tissues to evaluate clinical importance of SNP IL-28B examination in Hepatitis C treatment of Peg-IFN/RBV through SVR prediction model.Methods: This study consisted of two parts. First, a cross-sectional study on chronic Hepatitis C patients who completed Peg-IFN/RBV therapy. The second part was case control study on patients underwent liver biopsy and immunohistochemical staining.Results: Sustained virological response was significantly higher in CC allele of SNP IL-28B compared to non CC allele (p = 0.015). Higher expression of IFN-l3 was found in CC allele compared to non CC allele (p = 0.018). On the other hand, there is no significant difference between SVR and expression of IFN-l3 (p = 0.237) and IFN-l3 receptor (p = 0.237). Risk factor for SVR probability were formulated into p = 1 / (1 + e-y); e = 2.7, y = -2.498 + 2.652 (SNP IL-28B) + 2.029 (thrombocytes) for pretreatment while for on treatment risk factor y = -0.223 + 2.621 (RVR)Conclusion: SNP IL-28B was important pretreatment risk factor in genotype 1 chronic Hepatitis C treated with dual therapy. Major allele of IL-28B expressed more IFN-l3 and its receptor in response to HCV although no association between both factors was found. RVR was the best on treatment factor for SVR. Further evaluation study was required to find other possible factors affecting SVR achievement, Background: SNP IL-28B played an important role in achieving sustained virological response (SVR) among different ethnics in chronic Hepatitis C patients and is considered potential in predicting treatment response of Pegylated interferon/ribavirin (Peg-IFN/RBV) combination and spontaneous clearance in acute hepatitis. Up to date, molecular mechanism underlying correlation between SNP IL28B and SVR has not been fully understood yet although it is predicted to be related to IFN-λ3 and IFN-λ3 receptor in liver tissues.Aim: Understanding the association between SNP IL-28B and SVR in chronic Hepatitis C treatment and expression of IFN-l3 and IFN-l3 receptor in liver tissues to evaluate clinical importance of SNP IL-28B examination in Hepatitis C treatment of Peg-IFN/RBV through SVR prediction model.Methods: This study consisted of two parts. First, a cross-sectional study on chronic Hepatitis C patients who completed Peg-IFN/RBV therapy. The second part was case control study on patients underwent liver biopsy and immunohistochemical staining.Results: Sustained virological response was significantly higher in CC allele of SNP IL-28B compared to non CC allele (p = 0.015). Higher expression of IFN-l3 was found in CC allele compared to non CC allele (p = 0.018). On the other hand, there is no significant difference between SVR and expression of IFN-l3 (p = 0.237) and IFN-l3 receptor (p = 0.237). Risk factor for SVR probability were formulated into p = 1 / (1 + e-y); e = 2.7, y = -2.498 + 2.652 (SNP IL-28B) + 2.029 (thrombocytes) for pretreatment while for on treatment risk factor y = -0.223 + 2.621 (RVR)Conclusion: SNP IL-28B was important pretreatment risk factor in genotype 1 chronic Hepatitis C treated with dual therapy. Major allele of IL-28B expressed more IFN-l3 and its receptor in response to HCV although no association between both factors was found. RVR was the best on treatment factor for SVR. Further evaluation study was required to find other possible factors affecting SVR achievement]"

"Interferon (IFN) merupakan sitokin yang diproduksi oleh berbagai tipe sel sebagai respon rangsangan terhadap stimulasi virus, bakteri, parasit, sel tumor, atau antigen lain. Interferon α termasuk kelompok IFN tipe I yang mempunyai berbagai efek biologis yang meliputi antiviral, antitumor dan juga sebagai immunoterapetik. Penelitian ini bertujuan untuk mensintesis protein rekombinan human IFN α2a melalui sistem ekspresi pada bakteria E. coli BL21(DE3). Pada gen human ifn α2a dilakukan penambahan situs pemotongan enzim restriksi Nco I dan Xho I menggunakan metode PCR, kemudian dilanjutkan dengan proses ligasi ke vektor pET-32b(+) dan selanjutnya ditransformasikan pada E. coli DH5α.Hasil sekuensing menunjukkan bahwa vektor rekombinan (pET-32b(+)-IFN α2a) memiliki urutan nukleotida yang benar. Vektor rekombinan ini selanjutnya ditransformasikan ke dalam E.coli BL21(DE3). Klon transforman yang diperoleh dikultur dan diinduksi dengan penambahan IPTG 1 mM sehingga mengekspresikan protein rekombinan human IFN α2a. Dari hasil isolasi, diperoleh protein rekombinan human IFN α2a dalam bentuk protein terfusi sehingga mempermudah proses deteksi dan purifikasi. Protein dikarakterisasi melalui metode SDS PAGE dilanjutkan dengan Western blot dan pewarnaan CBB. Pita protein rekombinan human IFNα2a yang diperoleh berukuran 36 kDa. Hasil maksimal ditunjukkan ekspresi pada suhu 37⁰C dengan waktu inkubasi 5 jam setelah induksi.

---

Interferon (IFN) is a cytokine produced by various cell types as a response of stimulation to viruses, bacteria, parasites, tumor cells, or other antigens. Interferon α type I IFN groups have various biological effects, including antiviral, antitumor and immunotherapeutic. The aim of this research is to synthesize recombinant human IFN α2a proteins through bacterial expression systems in E. coli BL21 (DE3). Addition genes of human IFN α2a, which are restriction enzyme cutting sites for Nco I and Xho I, are added through PCR method. This step is followed by ligation process to the pET-32b(+) vector and then transformed into E. coli DH5α.The recombinant vector (pET-32b(+)-IFN α2a) has a nucleotide right sequence after it was being sequenced, after was transformed into E. coli BL21 (DE3). Obtained transformant clones were cultured and induced by addition of IPTG 1 mM to produce the expression of recombinant human IFN α2a proteins. As result of isolation process, recombinant protein of human IFN α2a are collected in fused protein thus can simplify the detection and purification method. The proteins are characterized by the SDS PAGE method followed by Western blot and CBB staining. The results show that the recombinant human IFN α2a protein bands are exactly 36 kDa. The maximum expression results were obtained at 37⁰C with 5 hours incubation after induction process."

"Pendahuluan: Imunoregulasi yang terjadi pada kehamilan menyebabkan ibu hamil lebih rentan terhadap infeksi, termasuk infeksi parasit. Salah satu parasit intestinal yang paling sering ditemukan di negara berkembang adalah Blastocystis-terutama banyak dijumpai pada populasi imunosupresi. Belum diketahui apakah infeksi Blastocystis dapat memengaruhi respon imun seluler terhadap infeksi patogen lain. Penelitian ini bertujuan untuk mengetahui hubungan infeksi Blastocystis pada kehamilan dengan respon imun seluler terhadap infeksi tuberkulosis, yang dimodelkan dengan stimulasi purified protein derivative (PPD). Metode: Penelitian ini menggunakan desain studi potong lintang dengan data yang bersumber dari penelitian utama yang telah dilakukan di daerah endemik Blastocystis. Sebanyak 98 ibu hamil trimester ketiga menjadi sampel dalam penelitian ini. Status infeksi Blastocystis ditetapkan berdasarkan pemeriksaan mikroskopis sampel feses. Respon imun seluler yang dinilai adalah kadar sitokin proinflamasi IFN-γ dan sitokin antiinflamasi IL-10 yang diambil dari kultur darah subjek dan diukur dengan Luminex assay.Hasil: Kadar IFN-γ dan IL-10 setelah stimulasi PPD lebih tinggi pada kelompok ibu hamil sehat dibandingkan ibu hamil terinfeksi Blastocystis, tetapi perbedaan kadar ini tidak signifikan untuk IFN-γ (p=0,356) dan signifikan untuk IL-10 (p=0,001). Perbandingan kadar sitokin setelah stimulasi PPD dengan basal yang dihitung dalam bentuk rasio menunjukkan hasil yang lebih tinggi pada kelompok ibu hamil sehat baik untuk IFN-γ dan IL-10, tetapi keduanya tidak bermakna secara statistik (p=0,428 untuk rasio IFN-γ dan p=0,564 untuk rasio IL-10). Rasio keseimbangan sitokin proinflamasi-antiinflamasi (rasio IFN-γ/rasio IL-10) pascastimulasi PPD lebih tinggi pada kelompok terinfeksi Blastocystis, meskipun tidak signifikan secara statistik (p=0,741). Kesimpulan: Infeksi Blastocystis pada ibu hamil tidak menunjukkan perbedaan respons imun terhadap stimulasi PPD dibandingkan dengan ibu hamil sehat.

---

Introduction: Predominant immunoregulatory state in pregnancy is associated with higher risk of infection, including parasitic infection. One of the most common intestinal parasites found in developing country is Blastocystis-which mainly found in immunocompromised population. It is not yet known whether Blastocystis infection could influence cellular immune response to other pathogens. Therefore, this research aims to discover the association between Blastocystis infection in pregnancy with cellular immune response to tuberculosis infection, which is modelled by purified protein derivative (PPD) stimulation. Method: This is a cross-sectional study which uses data from primary research that has been done in a Blastocystis-endemic area. Study samples consist of 98 pregnant women in their third trimester. Blastocystis infection was determined from microscopic examination of stool specimen. The cellular immune response is assessed by measuring serum level of IFN-γ and IL-10 as pro- and anti-inflammatory cytokine, respectively. The serum is obtained from a whole blood culture, and its cytokine level will further be measured with Luminex assay.Result: IFN-γ and IL-10 level with PPD-stimulation is higher in healthy pregnant women compared to Blastocystis-infected subjects, but this difference is not statistically significant for IFN-γ (p=0.356) and significant for IL-10 (p=0.001). The PPD-stimulated/basal ratio of both IFN-γ and IL-10 is also higher in healthy pregnant women, but it is not statistically significant (p=0.428 for IFN-γ ratio and p=0.564 for IL-10 ratio). Although not statistically significant, the pro- and anti-inflammatory cytokine balance (IFN-γ ratio/IL-10 ratio) after PPD stimulation is higher in pregnant women infected with Blastocystis (p=0.741). Conclusion: There is no difference in the cellular immune response to PPD stimulation in pregnant women infected with Blastocystis compared to healthy pregnant women."

"Pendahuluan. Penglepasan interferon-gamma oleh limfosit T yang antigenspecific akan meningkat setelah sel tersebut dipaparkan kembali dengan antigen tuberkulosis (TB) secara in vitro, khususnya apabila sel tersebut berasal dari lokasi infeksi TB aktif. Penelitian ini merupakan uji diagnostik pemeriksaan interferon-gamma release assay (IGRA) metode enzyme-linked immunospot (ELISPOT), yaitu T-SPOT.TB®, untuk deteksi TB pleura menggunakan spesimen sel mononuklear (MN) cairan pleura. Metode. Sebanyak 48 pasien efusi pleura terduga TB dengan karakteristik cairan pleura eksudatif berdasarkan kriteria Light dan dominasi sel MN lebih dari 50% dilakukan pemeriksaan T-SPOT.TB, biakan TB media cair Mycobacterial Growth Indicator Tube (MGIT), dan aktivitas adenosine deaminase (ADA) cairan pleura. Hasil. Dengan baku emas biakan TB MGIT didapatkan nilai sensitivitas 100%, spesifisitas 20%, nilai prediksi positif (NPP) 20%, dan nilai prediksi negatif (NPN) 100%. Dengan baku emas kombinasi biakan TB MGIT dan aktivitas ADA didapatkan nilai sensitivitas 100%, spesifisitas 88,89%, NPP 97,5%, dan NPN 100%. Kesimpulan. IGRA metode ELISPOT menggunakan spesimen cairan pleura merupakan pemeriksaan yang cepat dan bermanfaat sehingga dapat dipertimbangkan sebagai pemeriksaan tambahan pada pasien efusi pleura terduga TB.

---

Introduction. The release of interferon-gamma by antigen-specific T lymphocytes increases after rechallenge with tuberculosis (TB) antigen in vitro, especially at a localized site of TB infection. This study aimed to evaluate the diagnostic value of a commercial enzyme-linked immunospot (ELISPOT) assay for interferon-gamma, T-SPOT.TB®, in the diagnosis of TB pleurisy using pleural fluid mononuclear cells.

Methods. 48 subjects, presumed to have pleural TB with exudative pleural effusion by Light's criteria, dominated by mononuclear cells, had their pleural fluid specimen tested with T-SPOT.TB, TB Mycobacterial Growth Indicator Tube (MGIT) culture, and pleural fluid adenosine deaminase (ADA) activity.

Results. The sensitivity, specificity, positive and negative predictive values of the assay were 100%, 20%, 20%, 100%, respectively, if TB MGIT culture was used as the gold standard, and 100%, 88,89%, 97,5%, 100%, respectively, if TB MGIT culture and ADA activity of pleural fluid were used as the gold standard.

Conclusion. The ELISPOT assay for interferon-gamma is useful and rapid so it can be considered as a supplementary test to explore TB pleurisy."

"Infeksi virus hepatitis B (VHB) merupakan masalah kesehatan global. Terapi yang ada saat ini hanya berdampak minimal terhadap covalently closed circular deoxyribonucleic acid (cccDNA), sehingga eradikasi sulit dicapai. Matriks Metaloproteinase-9 (MMP-9) berperan dalam meningkatkan replikasi VHB, namun belum ada penelitian yang mengevaluasi peran penghambat MMP-9 terhadap replikasi VHB. Kultur primer hepatosit diperoleh dari Tupaia javanica dan diinfeksi dengan VHB manusia, kemudian dibagi menjadi dua kelompok, yaitu kontrol dan intervensi. Kelompok intervensi diberikan penghambat MMP-9 dosis 1 nM, 3 nM, dan 7 nM. Peripheral blood mononuclear cells (PBMC) manusia diperoleh dari pasien hepatitis B kronik, kemudian dibagi menjadi dua kelompok, yaitu kontrol dan intervensi. Kelompok intervensi diberikan penghambat MMP-9 dosis 3 nM. Dilakukan pengukuran HBsAg, DNA VHB, cccDNA, MMP-9, type-1 IFN receptor 1 (IFNAR1), dan interferon-β (IFN-β) sebelum dan 72 jam setelah pemberian intervensi pada kedua kelompok. Pada kultur primer hepatosit T. javanica yang diinfeksi VHB manusia, pemberian penghambat MMP-9 dosis 1 nM, 3 nM, dan 7 nM secara konsisten menurunkan kadar HBsAg, DNA VHB, cccDNA, dan MMP-9. Dosis 3 nM meningkatkan kadar IFNAR1, sedangkan dosis 7 nM meningkatkan kadar IFN-β. Dosis 3 nM menunjukkan efek yang lebih optimal dibandingkan dosis lainnya. Pada PBMC manusia dengan infeksi VHB, pemberian penghambat MMP- 9 dosis 3 nM menurunkan kadar HBsAg, DNA VHB, cccDNA, dan MMP- 9, serta meningkatkan kadar IFN-β, namun menurunkan kadar IFNAR1. Studi ini menunjukkan bahwa pemberian penghambat MMP-9 dapat menurunkan kadar HBsAg, DNA VHB, cccDNA, dan MMP-9, serta meningkatkan kadar IFN-β pada kultur primer hepatosit T. javanica dan PBMC manusia.

---

Hepatitis B virus (HBV) infection remains a significant global health concern. Current therapies have minimal impact on covalently closed circular deoxyribonucleic acid (cccDNA), making HBV eradication difficult. Matrix Metalloproteinase-9 (MMP-9) enhance HBV replication, but its inhibition has not been studied. Primary hepatocyte cultures were obtained from Tupaia javanica and infected with human HBV, then divided into control and intervention groups. The intervention groups were treated with MMP-9 inhibitors at doses of 1 nM, 3 nM, and 7 nM. Human peripheral blood mononuclear cells (PBMC) were isolated from chronic hepatitis B patients and divided into control and intervention groups. The intervention groups received MMP-9 inhibitors at dose of 3 nM. Measurements of HBsAg, HBV DNA, cccDNA, MMP-9, type-1 IFN receptor 1 (IFNAR1), and interferon-β (IFN-β) were performed before and 72 hours after intervention in both groups. In T. javanica primary hepatocyte culture infected with human HBV, MMP-9 inhibitors at doses of 1 nM, 3 nM, and 7 nM consistently decreased levels of HBsAg, HBV DNA, cccDNA, and MMP-9. The 3 nM dose increased IFNAR1 levels, while the 7 nM dose increased IFN-β levels. The 3 nM dose demonstrated the most optimal effects. In human PBMC with HBV infection, MMP-9 inhibitor at dose of 3 nM decreased levels of HBsAg, HBV DNA, cccDNA, MMP-9, increased IFN-β levels, but reduced IFNAR1 levels. This study shows that administration of MMP-9 inhibitors reduced levels of HBsAg, HBV DNA, cccDNA, and MMP-9, while increased IFN-β levels in T. javanica primary hepatocyte cultures and human PBMC."