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"Ruang Lingkup dan Cara Penelitian : Sindrom Down (SiD) merupakan suatu kelainan genetik yang paling sering dijumpai yang menyebabkan retardasi mental. SiD disebabkan oleh kelainan jumlah kromosom atau aberasi numerik yaitu trisomi 21 sehingga penderita mempunyai susunan kromosom 47,XX,+21 atau 47,XY,+21. Trisomi kromosom'21 disebabkan oleh proses gagal pisah ('nondisjunction') yang dapat terjadi baik pada ibu (75-80%) maupun pada ayah (20-25%). Beberapa studi yang berusaha mengungkap sebab-sebab atau etiologi dari gagal pisah telah dilakukan, namun penyebab gagal pisah kromosom 21 masih tetap banyak yang belum diketahui. Kemungkinan bahwa banyak faktor atau mekanisme turut berperan. Penelitian genetika molekuler ini bertujuan untuk melihat polimorfisme panjang fragmen restriksi (RFLP) DNA satelit alfa daerah sentromer kromosom 13/21 (D13Z1/D21Z1) yang mungkin berhubungan dengan proses terjadinya gagal pisah pada trisomi 21 yang menyebabkan SiD. Metode yang digunakan dalam penelitian ini adalah Blot Southern dengan menggunakan enzim restriksi EcoRI dan Xba I. Hasil dan Kesimpulan : 1. Dengan menggunakan enzim restriksi EcoRI dan pelacak DNA satelit alfa kromosom 13/21 (D13Z1/D21Z1) terdapat polimorfisme (RFLP) yang tidak spesifik pada wanita yang mempunyai anak SiD. RFLP terdapat pada fragmen 850, 1190, 1360 dan 1870 pb. 2. Dengan menggunakan enzim restriksi Xba I dan pelacak DNA satelit alfa kromosom 13/21 (D13Z1/D21Z1) tidak terdapat polimorfisme (RFLP) baik pada wanita yang mempunyai anak SiD maupun pada wanita pembanding. Hibridisasi menghasilkan 8 fragmen yang seragam pada semua individu yang diperiksa dengan ukuran: 0,7 ; 0,9 ; 1.0 ; 1,2 ; 1,4 ; 1,5 ; 1,7 dan 1,9 kb. Hal ini berarti bahwa dengan menggunakan enzim Xba I, polimorfisme DNA satelit alfa kromosom 13/21 tidak terdeteksi."

"Sampai saat ini di Indonesia belum diketahui prevalensi seropositif CMV pada darah donor sehingga belum dilakukan uji saring terhadap antibodi CMV dan analisis DNA CMV pada PRC leukodepleted secara rutin untuk kemanan darah donor. Tujuan penelitian ini adalah mendapatkan informasi efektifitas teknik leukodeplesi PRC terhadap deteksi DNA CMV, mendapatkan informasi mengenai prevalensi PRC dengan antibodi IgG CMV positif dan mendapatkan informasi mengenai prevalensi DNA CMV positif pada PRC non-leukodepleted serta PRC leukodepleted di UTD PMI Provinsi DKI Jakarta. Metode yang digunakan desain potong lintang cross sectional dengan jumlah sampel 113 darah donor yang telah memenuhi kriteria inklusi. Uji saring antibodi IgG CMV menggunakan metode indirect chemiluminescence immunoassay ChLIA dengan alat Liason XL 10050 Chemiluminescence Analyzer dan analisis DNA CMV menggunakan metode qPCR untuk deteksi UL54 CMV dengan alat Roche Light Cycler 480 II. Hasil penelitian menunjukkan prevalensi IgG CMV positif sebanyak 111 sampel 98,23 dan IgG CMV negatif sebanyak 2 sampel 1,77 . Prevalensi DNA CMV positif pada PRC non- leukodepleted adalah 1 sampel 0,88 dan PRC leukodepleted adalah 0 sampel 0. Kesimpulan penelitian ini, PRC leukodepleted efektif dalam meningkatkan keamanan darah donor terhadap infeksi CMV.Kata kunci: Cytomegalovirus, PRC Leukodepleted, IgG CMV, qPCR UL54 CMV.

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To date, the seropositive prevalence of CMV in blood donor is still remaining unknown. Therefore, no screening test for CMV antibody and CMV DNA analysis on leukodepleted PRC that is routinely performed in Indonesia for the safety of the blood donor.

The purpose of this study was to obtain information on the effectiveness of PRC leukodepleted techniques on CMV DNA detection, to obtain information on the prevalence of PRC with positive CMV IgG antibodies and to obtain information on the prevalence of positive CMV DNA in non leukodepleted PRC and leukodepleted PRC at UTD PMI DKI Jakarta.

Cross sectional design with total sample of 113 donor blood that has fulfilled the inclusion criteria was used as methodology. Indirect chemiluminescence immunoassay ChLIA method with Liason XL 10050 Chemiluminescence Analyzer was used for IgG CMV antibody screening test and qPCR technique with UL54 CMV by Roche light cycler 480 II was used for CMV DNA analysis.

The results showed that 111 samples 98.23 were positive to IgG CMV and 2 samples 1.77 was negative to CMV IgG. The prevalence of positive CMV DNA in PRC before leukodepleted was 1 sample 0.88 and PRC after leukodepleted was 0 sample 0.

The conclusion of this study is leukodepleted PRC effective to reduce the spread of CMV infection through blood transfusions.Key words Cytomegalovirus, PRC Leukodepleted, IgG CMV, qPCR UL54 CMV."

"Demam berdarah dengue merupakan salah satu penyalcit wabah di didunia yang telah merenggut banyak nyawa. Pada tahun 2004 tetjadi wabah demam berdarah dengue di Jakarta dengan lebih dati 10.000 kasus dengan angka kematian dilaporkan sebanyak 603 orang dengan kasus infeksi serotipo virus dengue terbanyak adalah serotipe 3. Penelitian ini menggueakan metode PCR. Pada penelitian ini dilakukan analisls variasi genetik genE, NSI, dan NS3 dari virus dengue tipe 3 yang diisolasi dari pasien dengan manifestasi klinis yang berbeda Strain DS2207 (DD), DS4607 (DBD), DSA0206 (DSS) yang diisolasi dari kasus dengue 2006-2007 dan dibandingkan dengan 12 strain yang berasal dati Indonesia, Thailand, Tahiti, Venezuela, Malaysia. Homologi etikleotida genE berkisar antara 92,4- 100 % sedangkan asam amino E 97- 100%. Homologi nukleotida gen NSI berkisar antara 93,4- 100 %, sedangkan asam amino NSl 97,2- 100%. Homologi nukleotida gen NS3 berkisar antara 92,8 - 99,5 % sedangkan asam amino NS3 98,2-99,7 % terdapat variasi pada ketiga gen, tapi belum ditemukan adanya perbedaan spesifik dari manifestasi klinis yang ditimbulkan.

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Dengue hemorrhagic fever is one of the epidemically disease that widely spread and has taken many lives. In the year of 2004, the epidemic of dengue hemorrhagic fever occurred in Jakarta with more than 10,000 number of cases, and 603 reported to death, where stereotype 3 showed as the most dengue virus stereotype infection cases. This research used the PCR (polymerase chain reaction) method, along with an analysis in genetically variety of E, NSI, and NS3 Gene of isolated Type 3 Dengue Virus (from the patient with any different clinical symptom). Strain of DS2207 (Dengue fever), 084607 (Dengue hemorrhagic fever), DSA0206 (Dengue shock syndrome) has been isolated of the 2006-2007 dengue cases, and compared' with 12 strain from Indonesia, Thailand, Tahiti, Venezuela, and Malaysia.

Nucleotide homology of the E gene is somewhere around 92,4% up to 100%, with theE aroino acid reached 97% up to 100%. Nucleotide homology of the NSl gene is somewhere around 93,4% up to 100%, with the NSI amino acid reached 92,2 % up to 100 % nucleotide homology of the NS3 gene is somewhere around 92,8 % up to 99,5 % with the NS3 amino acid reached 98,2 % up to 99,7 %. There were also variety of those genes, without any specipic differences among different manifestations."

"Legionellosis is a collection of infection that emerged in the second half of the 2Oth century, and that are caused by Legionella pneumophila and related bacteria. Legionellosis consists of two clinical syndromes, Legionnaires?disease is characterisized by pneumonia and pontiac fever is self-limiting, influenza like illness. Outb According this study, the sensitivity of duplex PCR to detect Legionella pneumophila in sterile NaCl 0.9% is 2.8 CFU/ml. The sensitivity of the duplex PCR in seeded water samples are 62 CFU/400ml of tap water sample, 32 CFU/400ml of sterile distiiled water and 32 CFU/400 ml of sterile NaCl 0.9%. The culture method in this study can not recovered Legionella from seeded water samples. The presence of Legionella .spp and Legionella pneumophila in cooling tower water was investigated using the duplex PCR. Of 9 cooling tower water sample and 3 tap water sample, 8 were positive for Legionella spp, 1 were positive for Legionella pneumophila and 3 were negative. According detection Legionella in seeded water samples and cooling tower water, the culture method can not be used to recover Legionella, but the duplex PCR can be used as rapid detection for Legionella spp and Legionella pneumophila.

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Legionella pneumophila merupakan penyebab utama lgionellosis yang mulai muncul pada pertengahan abad 20. Legionellosis dapat berkembang menjadi dua keadaan klinik, pertama Legionnares? disease yang merupakan penyakit multi sistem pneumonia, kedua Pontiac fever suatu penyakit mirip dengan flu dan dapat sembuh dengan sendirinya. Umumnya kasus legionellosis terjadi akibat dari kontaminasi pada sistem air panas maupun dingin pada gedung bertingkat seperti cooling tower, kondensor, spa, kolam renang, Oleh karena itu deteksi bakteri Legionellla pada sistem air di gedung bertingkat dan rumah sakit diperlukan untuk mencegah legionellosis nosokomial ataupun komunitas. Deteksi legionella dengan metode konvensional memerlukan media khusus dan waktu inkubasi yang lama. Pada penelitian ini duplex PCR dikembangkan untuk mendeteksi Legionella spp dan Legionella pneumophila pada sampel air cooling lower, dengan primer dari sekuens gen 16S rRNA unluk mendeteksi Legionella spp Serta primer sekuens gen nano untuk mendeteksi Legionella pneumophila. Pada penelitian ini Duplex PCR dapat digunakan untuk mendeteksi Legionella pneumophila dalam suspensi NaCl 0.9% hingga batas deteksi 2,8 CFU/ml. Hasil uji simulasi menggunakan sampel air yang ditambahkan pengenceran berseri Legionella pneumophila menunjukkan batas deteksi hingga 62 CFU/ 400 ml air kran, 32 CFU/400 ml akuadest steril dan 32 CFU/ 400 ml NaCl 0.9% stril. Hasil uji sirnulasi dengan metode kultur tidak menunjukkan pertumbuhan koloni pada agar BCYE plus. Hasil uji coba Duplex PCR terhadap 9 sampel air cooling tower dan 3 sampel air kran adalah satu sampel menunjukkan pita spesiiik L. pneumophia, 8 sampel yang menunjukkan pita spesifik Legionella spp dan 3 sampel negatif. Berdasarkan uji simulasi dan pemeriksaan sampel air cooling lower; metode kultur pada penelitian ini belum dapat mendeteksi keberadaan bakteri Legionella, sedangkan deteksi Legionella spp dan L. pneumophila dapat dilakukan dengan metode duplex PCR."

"Tuberkulosis sejak lama merupakan salah satu penyebab utama kematian manusia karena penyakit infeksi, terutama didaerah yang dilanda kemiskinan dan malnutrisi. Penyakit ini menyerang banyak organ pada tubuh manusia terutamanya adalah paru-paru. Peningkatan jumlah kasus tuberkulosis dipengaruhi oleh infeksi HIV dan resistensi terhadap berbagai macam kombinasi obat. Di Indonesia, infeksi M. tuberculosis oleh strain Beijing diyakini memiliki penyebaran yang paling luas dibandingkan dengan strain lainnya. BCG merupakan vaksin tunggal yang digunakan untuk pencegahan tuberkulosis, namun daya proteksi dan efikasinya berbeda-beda. Protein Mce1A merupakan protein yang diduga berperan penting pada hal invasi dan pertahanan M. tuberculosis didalam makrofag. Beberapa studi telah melakukan penelitian ini, namun di Indonesia belum pernah dilakukan penelitian mengenai ekspresi protein Mce1A Mycobacterium tuberculosis strain Beijing sebagai isolat lokal. Oleh karena itu, pada penelitian ini akan dilakukan pengklonaan dan ekspresi protein Mce1A Mycobacterium tuberculosis strain Beijing lokal dan strain standar H37Rv sebagai pembanding. Gen Mce1A M. tuberculosis strain Beijing dan H37Rv diamplifikasi dengan teknik PCR dan diinsersikan kedalam vektor pET28a. Escherichia coli BL21 kemudian ditransformasi dengan plasmid rekombinan tersebut. Protein Mce1A rekombinan diekspresikan dengan induksi IPTG. E. coli BL21 berhasil ditransformasi dengan plasmid rekombinan yang mengandung sisipan gen Mce1A dengan arah orientasi dan kerangka baca yang benar. Tidak ada mutasi yang ditemukan pada asam amino yang menjadi epitope pengenalan sel B dan sel T. Hasil ekspresi protein Mce1A pada E.coli BL21 menunjukkan pita protein yang lebih tinggi dari seharusnya. Konfirmasi keberadaan protein dilakukan menggunakan teknik Western Blot dengan anti-his detector. Protein Mce1A rekombinan yang telah berhasil diekspresikan pada E.coli BL21 diduga berada dalam bentuk dimer. Hal ini dapat digunakan sebagai data awal kondisi ekspresi untuk pengembangan vaksin subunit pada penelitian berikutnya.

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For past centuries until nowadays, tuberculosis remains the leading cause of death in the world from infectious disease wherever poverty, malnutrition and poor housing prevail. Tuberculosis is primarily a disease of the lungs, but may spread to other sites or proceed to a generalized infection. The wide spread of tuberculosis has been further aggravated by another infection disease such as HIV-AIDS and drug resistance. Many strain of Mycobacterium tuberculosis caused tuberculosis infection in Indonesia, but Beijing strain are the most. Bacille Calmette-Guerin (BCG) is the current vaccine for tuberculosis but it has different protection function and efficacy. According to function analysis, mce1A gene predicted has a role in host invasion by Mycobacterium tuberculosis and survival of the pathogen in human macrophages. Several studies abroad have done this research, but in Indonesia, study about protein expression of Mce1A gene of Mycobacterium tuberculosis Beijing strain as local isolates has not much being done. Therefore, in this study we will performed cloning and protein expression of Mce1A gene Mycobacterium tuberculosis Beijing strain as local isolate and standard strain H37Rv as a comparison on expression vector Escherichia coli BL21. Mce1A gene from M. tuberculosis Beijing and H37Rv strain was amplified by PCR and inserted in the vector pET28a. E. coli BL21 then transformed with the recombinant plasmid. Mce1A recombinant protein then expressed with IPTG induction. This study indicate that E. coli BL21 succesfully transformed with a recombinant plasmid containing the Mce1A gene insertion with correct orientation and reading frame. There is no mutation found in the amino acids sequence for B and T cell epitope. Mce1A expression in E. coli BL21 showed protein bands that higher than expected. The protein was confirmed with western blotting using anti-his detector. We assume that Mce1A recombinant protein that have been expressed in E. coli BL21 is in dimeric form. This explanation should be valuable in further studies of expression at the protein level and exposure of proteins on the cell surface of M. tuberculosis under different experimental conditions."

"[ABSTRAKLatar belakang. Hepatitis B merupakan salah satu masalah kesehatan yang serius, diperkirakan lebih dari 2 milyar orang didunia telah terinfeksi virus hepatitis B (VHB). Dari jumlah ini kira-kira 360 juta orang mengalami infeksi khronis. Kematian terutama disebabkan karena sirosis hepatis dan karsinoma hepatoseluler. Salah satu upaya pencegahan penularan infeksi VHB adalah uji saring darah donor terhadap hepatitis B surface antigen (HBsAg) yang merupakan pemeriksaan skrinning yang dilakukan oleh Unit Transfusi Darah (UTD) di negara berkembang seperti Indonesia. Banyak peneliti membuktikan bahwa darah HBsAg negatif masih berpotensi menularkan infeksi VHB. Untuk itu meningkatkan keamanan darah, beberapa negara menambahkan parameter pemeriksaan antibodi terhadap hepatits B core (anti-HBc) sebagai petanda paparan terhadap infeksi VHB dan pemeriksaan antibodi terhadap hepatitis B surface (anti-HBs) sebagai tanda respon imun terhadap infeksi VHB. Dengan berkembangnya teknologi biologi molekuler, masa jendela infeksi VHB dengan seronegatif dapat di ketahui lebih cepat melalui deteksi DNA VHB dengan metode Nucleic Acid Test (NAT) multipleks yang dilanjutkan dengan NAT discriminatory. Beberapa penelitian terdahulu menunjukkan, didapatnya DNA VHB pada spesimen darah donor yang seronegatif dengan metoda NAT. Darah dengan HBsAg negatif, DNA VHB positif dengan atau tanpa anti-HBc dan atau anti-HBs merupakan darah asal donor dengan Hepatitis B Occult (HBO). Prevalensi donor dengan HBO di Indonesia berkisar antara 8- 10%.Metodologi. Penelitian ini menggunakan desain potong lintang (cross sectional) yang dilakukan di UTDP dan Lembaga Biologi Molekul Eijkman, dengan jumlah sampel 4.973 asal subyek donor darah dari 4 UTD daerah DKI, Kota Tanggerang,kota Depok dan Kabupaten Tanggerang. Terhadap sampel penelitian dilakukan pemeriksaan serologis HBsAg,anti-HBc,anti-HBs, NAT, dan PCR kuantitatif dan kualitatif, selanjutnya pada sampel yang HBsAg negatif, NAT positif dan didapatkan hasil PCR kuantitatif positif dilakukan pemeriksaan lanjutan sequencing asam amino untuk mengetahui ada tidaknya mutan HBsAg penyebab lolosnya deteksi serologi HBsAg oleh reagensia HBsAg yang digunakan untuk uji saring darah donor.Hasil. Didapatkan hanya 20 subjek (0,40%) mempunyai hasil HBsAg negatif dan NAT positif multipleks, dan hanya 16 subyek (80%) HBsAg negatif dan NAT discrimenatory positif. Lebih lanjut hasil pemeriksaan anti-HBc negatif dan anti- HBs positif/negatif didapatkan hanya1 subyek (6,25%), anti-HBc positif dan anti- Hbs negatif didapatkan 9 subyek (56,25%), hasil pemerksaan anti-Hbc dan anti- HBs positif 5 subyek (31,25%). Lebih lanjut dilakukan pemeriksaan PCR kualitatif dan didapatkan 3 subyek (18,75%) tidak terdeteksi, , 6 (37,5%) subyek menunjukkan hasil viral load yang low detection (dibawah sensitivitas alat), dan 7 subyek (43,75%) menunjukkan hasil viral load dapat di ketahui. Pada pemeriksaan PCR kualitatif dan sequencing didapatkan 2 subyek (28,57%) ditemukan mutasi pada gen S pada posisi 143 dimana terjadi subsitusi asam amino T143M.

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ABSTRACTBackground. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection.And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg.Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donorsResult. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.;Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection.And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg.Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donorsResult. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.;Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection.And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg.Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donorsResult. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.;Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection.And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg.Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donorsResult. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.;Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection.And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg.Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donorsResult. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution., Background. Hepatitis B is one of the most serious health problem. It is estimated that more than 2 bilion people have been infected by this virus, of which 360 milion are chronically infected with severe and fatal risk especially of cirrosis and hepatocellular carcinoma. One of the main ways to prevent transfusion transmitted HBV infection is blood screening for HBsAg. However, many studies have proven that HBsAg negative blood can still be infection. Therefore to enhance blood safety same countries have added antibody parameters in blood screening of donors – antibody for hepatitis B core antigen (anti-HBc) as marker for HBV infection and antibody for hepatitis B surface antigen (anti-HBs) as marker for immunological response to HBV infection.And with the development of molecular biology technology, HBV infection can be knowing faster in seronegative windows period with HBV DNA examination inspection by methods Nucleic Acid Test (NAT) multiplex and discrimenatory. In fact the results of seronegative blood is still there HBV virus with NAT, and this result we can called with the Occult hepatitis B (HBO). Prevalence of donors with HBO ranges 8- 10% in Indonesia. This study aims to determine the efectivity analized by NAT method blood donor with HBO to see continuity with the examination of anti-HBc, anti-HBs, viral load, and the cause of the HBO that mutations in the gene encoding HBsAg.Methodology. This study used a cross-sectional design (cross-sectional) conducted in UTDP and Eijkman Institute for Molecular Biology, with a sample of 4,973 sampless blood bloods donor from 4 units of blood transfusion area of Jakarta, Tangerang City, Depok and Bekasi district. Against sample serological examination HBsAg, anti-HBc, anti-HBs, NAT, quantitative and qualitative PCR, then samples were HBsAg negative, positive NAT and quantitative PCR positive results obtained further investigation amino acid sequencing to determine whether there is a mutant HBsAg cause serological detection of HBsAg escape by HBsAg reagents used for screening of blood donorsResult. There was 20 samples (0.40%) had results of HBsAg negative and positive NAT multiplex, and only 16 samples (80%) HBsAg negative and positive discrimenatory NAT. Furthermore, the results of the examination of anti-HBc and anti-HBs negative positive / negative obtained only 1 samples (6.25%), anti-HBc positive and negative anti-Hbs obtained 9 samples (56.25%), and anti-HBc , anti- HBs positive are 5 samples (31.25%). and then , qualitative PCR examination and had 3 samples (18.75%), is not detected, 6 (37.5%) samples, and some samples showed a low viral load results detection (sensitivity under tools value), and 6 samples (43.75%) shows viral load is positive. In qualitative PCR and sequencing obtained 2 samples (28.57%) found a mutation in the S gene at position 143 where there T143M amino acid substitution.]"

"Spag11a diketahui terekspresi secara spesifik pada kaput epididimis sehingga dimungkinkan protein tersebut memiliki fungsi yang spesifik untuk maturasi spermatozoa. Studi peran SPAG11A dalam maturasi spermatozoa di epididimis memerlukan produksi protein SPAG11A untuk dikarakterisasi. Tujuan dari penelitian ini adalah untuk mengklon, mengekspreesikan, dan mengkarakterisasi sifat antimikroba dari protein rekombinan SPAG11A. Insert cDNA Spag11a yang dihasilkan melalui PCR diklon ke dalam vektor pET100/D-TOPO. Plasmid rekombinan kemudian diekspresikan ke Escherichia coli BL21 DE3 star. Deteksi dari fusi protein rekombinan dilakukan dengan SDS-PAGE dan Western Blotting. IMAC Immobilized Metal Affinity Chromatography digunakan untuk mempurifikasi protein rekombinan. Uji antimikroba protein rekombinan dianalisis melalui pengukuran Optical density.PCR amplifikasi dari cDNA kaput epididimis mencit menghasilkan insert Spag11a berukuran 210bp. Insert tersebut kemudian dikloning ke dalam pET100/D-TOPO menghasilkan 1 rekombinan plasmid dari 10 koloni yang diskrining. Ekspresi rekombinan klon ke dalam E.coli BL21 menghasilkan fusi protein setelah diinduksi IPTG selama 4 jam. Fusi protein dikonfirmasi menggunakan Western Blotting menggunakan antibodi yang mengenali N-terminal His-Tag 21kDa dan protein SPAG11A. Uji antimikroba protein rekombinan SPAG11A mununjukkan tidak ada inhibisi yang signifikan terhadap laju pertumbuhan E.coli dan Bacillus subtilis. Insert Spag11a yang berukuran 210bp berhasil diklon ke dalam vektor pET100/D-TOPO. Ekspresi rekombinan Spag11a menghasilkan fusi protein berukuran 21kDa. Protein rekombinan SPAG11A tidak membawa sifat antimikroba terhadap E.coli dan B. subtilis.

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Spag11a is known to be specifically expressed in the caput region of the epididymis suggesting a specific function for sperm maturation. Study of SPAG11A role in the epididymal sperm maturation requires generating SPAG11A protein for characterization. The objective of this study was to clone, express and characterize antimicrobial property of the recombinant SPAG11A. Spag11a cDNA insert was generated by PCR and cloned in TOPO vector. Recombinant DNA plasmid was subsequently expressed in E coli BL 21 star. Detection of recombinant fusion protein was carried out using SDS PAGE and western immunobloting. IMAC Immobilized Metal Affinity Chromatography was used to purify recombinant protein. Optical density measurement was used to analyse antimicrobial property of the recombinant protein. PCR amplification of mouse caput epididymis cDNA produced a 210 bp insert of Spag11a. Cloning of the insert into TOPO pET100 resulted in 2 recombinants out of 10 colonies that were screened. Expression of recombinant clones in the E coli BL21 produced a fusion protein after being induced IPTG for 4 hours. Fusion protein was confirmed by western immunobloting using two antibodies recognizing N terminal His Tag 21 kDa and SPAG11A protein. Antimicrobial assay for SPAG11A recombinant showed no significant inhibition towards growth rates of E coli and Bacillus subtilis. A 210 bp Spag11a insert was successfully cloned into TOPO pET100 vector. Expression of recombinant spag11a produced a fusion protein of 21 kDa. SPAG11A recombinant protein does not have antimicrobial property towards E coli and B subtilis."

"ABSTRAKDNA eksogen merupakan DNA asing yang diintroduksi ke dalam sel dan sebagai dasar pengembangan vaksin DNA. Ekspresi gen pada DNA eksogen masih rendah dalam antigen presenting cell APC yang merupakan target utama dalam penerapan vaksin DNA, seperti monosit. Salah satu cara dalam meningkatkan ekspresi gen pada DNA eksogen adalah menyisipi sekuen internal inisiasi translasi yaitu IRES HIV-1. Oleh karena itu, pada penelitian ini dilakukan penambahan sekuen IRES HIV-1 pada hulu gen DNA eksogen yang akan diekspresikan di sel monosit manusia. DNA IRES HIV-1_eGFP diamplifikasi dari pcDNA5FRT/TO dan disubkloning ke pcDNA3.1 . DNA ditransfeksi ke kultur primer monosit dari darah manusia sehat. Sel berfluoresen, persentase sel, dan intensitas fluoresen diamati masing-masing dengan mikroskop fluoresen, Tali cytometer, dan Glomax. Plasmid pcDNA3.1_IRES HIV-1_eGFP berhasil dikonstruksi dan gen egfp berhasil diekspresikan pada sel monosit manusia. Persentase monosit berfluoresen dibandingkan sel kontrol meningkat dari 5 menjadi 11,54 kelompok pcDNA3.1_eGFP dan 12,9 kelompok pcDNA3.1_IRES HIV-1_eGFP p=0,297 . Intensitas fluoresen eGFP pada kelompok dengan IRES HIV-1 dalam total sel monosit 2,33 dan per monosit 0,069 meningkat signifikan dibandingkan kelompok pcDNA3.1_eGFP dalam total sel monosit 0,08 dan per monosit 0,001 p=0,001 . Oleh karena itu, penambahan IRES HIV-1 terbukti mampu meningkatkan ekspresi gen pada sel monosit secara signifikan.

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ABSTRACTExogenous DNA is an transient DNA that is introduced into cells and as a basis for developing DNA vaccines. Gene expression in exogenous DNA is still low in antigen presenting cells APC , which is a major target in the application of DNA vaccines, such as monocytes. One way to increase the expression of gene in exogenous DNA is to insert the internal sequence of translation initiation such as IRES HIV 1. Therefore, in this research, the addition of IRES HIV 1 sequence in the upstream gene of exogenous DNA to be expressed in human monocytes cells. IRES HIV 1 eGFP amplified from pcDNA5FRT TO and subcloned to pcDNA3.1 .. DNA were transfected into the primary culture of monocytes from healthy human blood. Fluorescent cells, cell percentages, and fluorescent intensity were observed with fluorescent microscope, Tali cytometer, and Glomax respectively. The pcDNA3.1 IRES HIV 1 eGFP successfully constructed and transfected in human monocytes cells. The percentage of fluorescent monocytes compared with control cells increased from 5 to 11.54 pcDNA3.1 eGFP group and 12.9 pcDNA3.1 IRES HIV 1 eGFP p 0.297 . The intensity of fluorescent eGFP in the group with IRES HIV 1 in total monocyte 2.33 and each monocyte 0.069 increased significantly compared to pcDNA3.1 eGFP group in total monocyte cell 0.08 and per monocyte 0.001 p 0.001 . Therefore, the addition of IRES HIV 1 has been shown to increase gene expression in monocyte cells significantly "

"ABSTRAKXanthin oksidase (XO) adalah enzim yang berperan dalam degradasi purin danpada aktivitasnya menghasilkan senyawa oksigen reaktif (reactive oxygen species,ROS). Penelitian pada cedera iskemia reperfusi membuktikan bahwa aktivitasenzim ini berperan sebagai penghasil utama ROS. Pada cedera iskemia reperfusiatau pada cedera hipoksia dan reoksigenasi (H/I) terjadi peningkatan aktivitas XO.Peran CoCl2 sebagai agen hipoksia mimikri telah banyak diketahui. PaparanCoCl2 menyebabkan respons yang menyerupai keadaan hipoksia melaluistabilisasi protein hypoxia-inducibel factor-1α dan juga melalui produksi ROS.Apakah induksi CoCl2 dapat meningkatkan aktivitas XO seperti halnya cederaiskemia reperfusi,merupakan pertanyaan pada penelitian ini. Tujuan daripenelitian ini adalah untuk mengetahui aktivitas XO pada jaringan hati dan ginjaltikus yang diinduksi CoCl2. Oleh karena itu dilakukan penelitian aktivitas XOpada jaringan hati dan ginjal tikus yang diinduksi CoCl2 intraperitoneal dengandosis 30 mg/kg BB dengan lama paparan 0 jam (sebagai kontrol); 2 jam, 8 jamdan 24 jam. Pada penelitian ini juga dilakukan pengukuran aktivitas antioksidankatalase yang berperan dalam menetralkan H2O2 sebagai hasil samping degradasipurin oleh XO. Hasil penelitian menunjukkan bahwa terdapat perbedaan aktivitasXO antara jaringan hati dan ginjal. Pada jaringan hati terjadi peningkatanaktivitas XO yang signifikan pada hipoksia mimikri 24 jam, sementara padajaringan ginjal tidak terjadi perubahan aktivitas XO. Aktivitas katalase keduajaringan memperlihatkan pola yang sama berupa penurunan aktivitas, walaupunpada ginjal secara statistik tidak signifikan, sementara penurunan aktivitaskatalase terendah terjadi pada jaringan hati dengan kondisi mimikri hipoksia 24jam. Ternyata tidak ada korelasi antara aktivitas XO dan katalase pada jaringanhati dan ginjal tikus yang diinduksi CoCl2.

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ABSTRACTXanthin oxidase (XO) is an enzyme that plays a role in the degradation of purineand the activity produces reactive oxygen species (ROS). Researches on ischemiareperfusioninjury have been proved that this enzyme acts as a major producer ofROS. In ischemia-reperfusion injury or hypoxic-reoxigenation injury (H/I) there isan increase in XO activity. CoCl2 has been widely known as an agent of hypoxiamimicry. CoCl2 exposure causes a response that resembles hypoxia conditionmediated by stabilization of hypoxia-inducible factor-1α protein and through theproduction of ROS also. Is CoCl2 induction can increase the activity of XO aswell as in ischemia reperfusion injury, remain unclear. The aim of this study wasto determine the activity of XO in the liver and kidney tissue of rats induced byCoCl2. The study was conducted in rats induced by intra peritoneal injection ofCoCl2 with a dose of 30 mg/kg BW with duration of exposure from 0 hrs (as acontrol); 2; 8; and 24 hrs. This study also measured the antioxidant activity ofcatalase that neutralized H2O2 as a by-product of purine degradation by XO.Results showed that there was a difference between XO activity in the liver andkidney tissue. In liver tissue XO activity was increased significantly in 24 hrshypoxia mimicry, while in kidney tissue, there were no changes in the activity ofXO. In both tissues catalase activity showed a similar pattern of decreasedactivity, although the changes of catalase activity in kidney was not statisticallysignificant, while in the liver tissue, the decrease of catalase activity was lowest in24 hrs hypoxia mimicry condition. Apparently there is no correlation between XOand catalase activity in the liver and kidney of rats induced by CoCl2."

"Expression of Virus Like Particles (VLP) containing the Influenza A haemagglutinin (HA) and neuraminidase (NA) may require simultaneous expression ofthe genes that encodes the proteins in order to obtain a balanced composition of I-IA and NA molecules in the VLP for optimal induction of protective immune response due to the increase of NA molecules in the VLP vaccine preparation. Such a condition can be more easily achieved by placement of the genes in the same vector DNA. A DNA construct for simultaneous expression of H5N1 Irriluenza A HA and NA proteins was thus constructed utilizing a pre constructed vector as the initial backbone DNA, which was a pcDNA3.I/His A plasmid that contained an insertion of the H5N1 NA gene. The sequences of internal ribosomal entry site (IRES) belonging to Murine Leukemia Virus (MLV) and Human Immrmodeficiency Virus (HIV), the HA gene of H5N1 influenza A, and the bsf gene of HIV IRES were introduced into the backbone DNA, such that the resulting DNA contains the MLV IRES upstream to the NA gene and the HIV IRES upstream to the HA gene, while the bsf gene was placed in between the NA gen and the HIV IRES. The IRES sequences were introduced for simultaneous expression of the HA and NA genes during the G2/ Mitosis phase of the cell cycle, while the bsf gene was introduced to prevent reinitiation process after termination of translation. The orientation and the accuracy of the nucleotide sequences of the inserted DNA fragments were analyzed using PCR and nucleotide sequencing. A clone that bears the inserted DNA fragments in the correct orientations with nucleotide sequences that have been confirmed by nucleotide sequencing to support proper expression of the HA and NA proteins, was obtained.

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Ekspresi Virus Like Particles (VLP) mengandung haemagglutinin (HA) dan neuraminidase (NA) Iniluenza A mungkin memerlukan ekspresi gen yang mengkode protein secara simultan supaya memperoleh komposisi molekul HA dan NA dalam VLP untuk induksi optimal respon imun protektif karena peningkatan molekul NA dalam penyiapan vaksin VLP. Kondisi tersebut dapat dengan mudah dicapai melalui penempatan gen dalam vektor DNA yang sama. Rancangan DNA untuk ekspresi protein HA dan NA Influenza H5Nl, kemudian dibuat menggunakan vektor pra konstruksi sebagai DNA backbone awal, yaitu plasmid pcDNA3.l/His A yang mengandung gen sisipan NA HSN1. Sekuen internal ribosomal entry site (IRES) Murine Leukemia Vims (MLV) dan Human Immimodeiicieney Virus (HIV), gen HA Influenza A HSN1, dan gen bsf-IRES HIV dimasukkan ke dalam DNA backbone, sehingga menghasilkan DNA yang mengandung IRES MLV pada hulu gen NA dan IRES HIV pada hulu gen HA, sedangkan gen bsf terletak di antara gen NA dan IRES HIV. Sekuen IRES dimasukkan untuk ekspresi gen HA dan NA secara simultan selama fase G2 / mitosis dalam siklus sel, sedangkan gen bsf dimasukkan untuk menoegah proses reinisiasi setelah terminasi translasi. Orientasi dan akurasi sekuen nukleotida iiagmen DNA sisipan dianalisis menggunakan pobrmerase chain reaction (PCR) dan sequencing nukleotida. Klona yang membawa fragmen DNA sisipan dengan selcucn nukleotida dalam orientasi benar telah dikonfirmasi melalui sequencing nukleotida untuk mendukxmg kebeuaran ekspresi protein HA dan NA yang diperoleh."