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Detection of INH Target Genes on DNA of Positive Acid Fast Stain Sputum Using PCR Technique R, Maria Lina; Bela, Budiman; Syaifudin, Mukh
Journal of the Indonesian Medical Association Vol. 57 No. 8 August 2007
Publisher : Journal of the Indonesian Medical Association

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Abstract

Tuberculosis, the world’s major diseases, is one of the emerging infectious diseases . The tuberculosis problem has become complicated and burdensome due to the emergence of drug resistan such as isoniazid (INH) resistant strains of Mycobacterium tuberculosis. The resistance can be detected by analyzing the genes which code the INH target genes such as kat-G and kas-A. Research to investigate the existence of the genes has being carried out on 25 positive acid fast stain sputum samples. The extraction of sputum DNA was done by means of three kind of methods including Boom’s method, proteinase-K, and boiling methods. PCR technique with two pairs of the oligonucleotide primers for kat-G and kas-A genes, subsequently continued with the agarose gel electrophoresis process, were used for detecting the existence of the genes. Results revealed that the existence of kat-G gene can be detected on 24 of the 25 sputum samples DNA which extracted by either Boom’s or proteinase-K methods, 1 sample were negative result, and by using boiling method, 3 samples showed the negative results. However, the DNA bands of 1 and 5 samples from 25 positive samples mentioned above, appeared to be thin bands by using Boom’s and proteinase- K, respectively. The existence of kas-A gene showed the positive results on 24 samples of the 25 sputum samples DNA and 1 sample was negative by means of Boom’s method, whereas with proteinase-K and boiling methods , the negative results appeared on 7 and 16 sputum samples, respectively. From the results mentioned above, it can be concluded that the best DNA extraction method is Boom’s method. However, the more simple method that is as same as Boom’s method, is necessary to be carried out.Keywords: PCR, kat-G gene, kas-A gene, INH, acid fast stain bacilli, Boom’s method, proteinase- K method, boiling method
SUPEROKSIDA DISMUT ASE (SOD) : APA DAN BAGAIMANA PERANANNYA DALAM RADIOTERAPI Nurhayati, Siti; Kisnanto, Teja; Syaifudin, Mukh
Buletin Alara Vol 13, No 2: Desember 2011
Publisher : BATAN

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UJI PRAKLINIS 99mTc-KANAMISIN SEBAGAI RADIOFARMAKA UNTUK PENCITRAAN INFEKSI Halimah, Iim; Ridwan, Ahmad; Syaifudin, Mukh
Jurnal Sains dan Teknologi Nuklir Indonesia Vol 16, No 1 (2015): Februari 2015
Publisher : BATAN

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ABSTRAK UJI PRAKLINIS 99mTc-KANAMISIN SEBAGAI RADIOFARMAKA UNTUK PENCITRA-AN INFEKSI. 99mTc-kanamisin merupakan salah satu radiofarmaka yang digunakan untuk mendiagnosis infeksi hingga ke bagian tubuh yang sangat dalam. Penelitian ini bertujuan untuk mendapatkan informasi mengenai karakteristik praklinis 99mTc-kanamisin meliputi toksisitas, sterilitas, pirogenitas, dan biodistribusi. Uji toksisitas dilakukan pada 5 ekor mencit yang diinjeksi 99mTc-kanamisin secara intra vena ekor, dilanjutkan dengan pengamatan sampai dengan 24 jam setelah injeksi. Hasil pengamatan menunjukkan bahwa 99mTc-kanamisin tidak bersifat toksik. Uji sterilitas dengan metode inokulasi 99mTc-kanamisin secara langsung pada medium nutrient agar dan tioglikolat cair menunjukkan bahwa 99mTc-kanamisin bersifat steril. Uji pirogenitas pada 3 ekor kelinci yang diinjeksi 99mTc-kanamisin secara intra vena pada telinga menunjukkan bahwa suhu total respon sebesar 2,9 oC, yang berarti 99mTc-kanamisin belum bebas pirogen. Biodistribusi 99mTc-kanamisin dilakukan pada mencit yang tidak diinfeksi dan yang diinfeksi dengan bakteri Escherichia coli dan Staphylococcus aureus secara intra-muskular pada 24 jam sebelum injeksi 99mTc-kanamisin. Beberapa sampel organ dan jaringan mencit diambil pada interval waktu 30, 60, dan 180 menit pasca injeksi 99mTc-kanamisin secara intravena melalui ekor mencit, menunjukkan bahwa 99mTc-kanamisin terakumulasi di dalam organ target yaitu otot paha kiri. Nilai rasio otot paha kiri terhadap otot paha kanan yang diperoleh sebesar 3,63 dan 5,64, masing-masing untuk E. coli dan S. aureus. Radiofarmaka 99mTc-kanamisin bersifat tidak toksik, steril, mengandung pirogen, terdistribusi di dalam tubuh mencit dengan baik, dan diekskresikan secara cepat dari dalam tubuh mencit melalui ginjal mulai 30 menit pasca injeksi, baik pada mencit yang diinfeksi bakteri maupun mencit yang tidak diinfeksi bakteri. ABSTRACT99mTc-KANAMYCIN PRECLINICAL TESTING AS A  RADIOPHARMACEUTICAL FOR INFECTION IMAGING. Infectious disease is the leading cause of death in worldwide, especially in developing countries such as Indonesia. Early detection and determination of the exact location of infection by imaging methods can facilitate treatment. 99mTc-kanamycin is one of the radiopharmaceuticals that widely used for such purpose. The aim of the study was to obtain the information on the preclinical characteristics of 99mTc-kanamycin including toxicity, sterility, pirogenicity, and biodistribution. Toxicity test conducted in 5 mice injected radiopharmaceutical 99mTc-kanamycin intra venous tail, showed that the radiopharmaceutical was not toxic for 24 hours after intravenous injection. Sterility testing of radiopharmaceutical 99mTc-kanamycin conducted with direct inoculation on Nutrient Agar and liquid Thioglicolat medium showed that the radiopharmaceutical was sterile. Pyrogenicity test conducted in 3 rabbits injected with the radiopharmaceutical 99mTc-kanamycin showed that the total temperature response was 2.9oC, that means that the radiopharmaceutical was not free from pyrogen. Biodistribution of 99mTc-kanamycin at intervals of 30, 60, and 180 minutes post intravenously injection through the tail of mice, infected with Escherichia coli and Staphylococcus aureus intramuscularly 24 hours earlier, showed that 99mTc-kanamycin accumulates in the target organ of the left thigh muscle. The ratio values of left to right thigh muscle were 3.63 and 5.64 for  E. coli and S. aureus, respectively. Radiopharmaceutical 99mTc-kanamycin was not toxic, sterile, pyrogen, distributed in the body of mice, and were rapidly excreted from the body through the kidneys starting 30 minutes post injection, both in bacterials infected or without bacterials infected mice. ABSTRACT 99mTc-KANAMYCIN PRECLINICAL TESTING AS A RADIOPHARMACEUTICAL FOR INFECTION IMAGING. Infectious disease is the leading cause of death in worldwide, especially in developing countries such as Indonesia. Early detection and determination of the exact location of infection by imaging methods can facilitate treatment. 99mTc-kanamycin is one of the radiopharmaceuticals that widely used for such purpose. The aim of the study was to obtain the information on the preclinical characteristics of 99mTc-kanamycin including toxicity, sterility, pirogenicity, and biodistribution. Toxicity test conducted in 5 mice injected radiopharmaceutical 99mTc-kanamycin intra venous tail, showed that the radiopharmaceutical was not toxic for 24 hours after intravenous injection. Sterility testing of radiopharmaceutical 99mTc-kanamycin conducted with direct inoculation on Nutrient Agar and liquid Thioglicolat medium showed that the radiopharmaceutical was sterile. Pyrogenicity test conducted in 3 rabbits injected with the radiopharmaceutical 99mTc-kanamycin showed that the total temperature response was 2.9oC, that means that the radiopharmaceutical was not free from pyrogen. Biodistribution of 99mTc-kanamycin at intervals of 30, 60, and 180 minutes post intravenously injection through the tail of mice, infected with Escherichia coli and Staphylococcus aureus intramuscularly 24 hours earlier, showed that 99mTc-kanamycin accumulates in the target organ of the left thigh muscle. The ratio values of left to right thigh muscle were 3.63 and 5.64 for E. coli and S. aureus, respectively. Radiopharmaceutical 99mTc-kanamycin was not toxic, sterile, pyrogen, distributed in the body of mice, and were rapidly excreted from the body through the kidneys starting 30 minutes post injection, both in bacterials infected or without bacterials infected mice.
PENERAPAN EFEK INTERAKSI RADIASI DENGAN SISTEM BIOLOGI SEBAGAI DOSIMETER BIOLOGI Lusiyanti, Yanti; SYAIFUDIN, MUKH
Jurnal Forum Nuklir JFN Vol 2 No 1 Mei 2008
Publisher : BATAN

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AbstrakInteraksi radiasi pengion dengan sistem biologi dapat menyebabkan berbagai macam efek biologik yang akan dimanifestasikan baik pada tingkat seluler, sitogenetik maupun tingkat molekuler. Berbagai macam metode biologik yang dimaksudkan untuk memperkirakan dosis radiasi telah dikembangkan oleh banyak peneliti menggunakan efek tersebut terutama dalam hal teIjadinya peristiwa kecelakaan radiasi. Hal ini dipertegas lagi dengan kenyataan bahwa dosimetri fisik tidak dapat diandalkan secara sendirian. Dengan kelebihan dan kekurangannya masing-masing, ulasan ulang ini memberikan gambaran yang meluas akan pentingnya uji atau biomarker dalam dosimetri biologi seperti kromosom disentrik, mikronuklei, fragmen kromosom,biokimia darah dan spermatogenesis. Adapun sampel biologik yang dapat dipergunakanuntuk pengkajian dosis radiasi yang diterima oleh pekeIja maupun korban kecelakaanantara lain darah, sperma, rambut, dan urin.Kata kunci : Interaksi radiasi pengion, sistem biologiAbstractInteraction between ionizing radiation with biological system could results in various types of biological effects which will be manifested either in cellular, cytogenetics or molecular levels. Various types of biological methods with the aim for predicting radiation dose have been developed by many researchers by using these effects mainly in the case of radiation accident. This case is supported by the fact that physical dosimetry could not available as alone. With their own advantages and disadvantages, this review provides a broader figure about the importance of assays or markers in biological dosimetry such as dicentric chromosome, micronuclei, chromosomal fragment, biochemistry of blood and spermatogenesis. Whereas the biological samples that can be used for assessing the radiation dose received by workers or accidental victims are blood, sperm, hair, and urine.Keywords: Interaction between ionizing radiation, biological system
UJI PROFIL PROTEIN KELENJAR LUDAH Anopheles sp. TERINFEKSI P. berghei PASCA IRADIASI GAMMA DENGAN TEKNIK SDS-PAGE UNTUK PENGEMBANGAN VAKSIN MALARIA Tetriana, Devita; Syaifudin, Mukh
Jurnal Sains dan Teknologi Nuklir Indonesia Vol 15, No 1 (2014): Februari 2014
Publisher : BATAN

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UJI PROFIL PROTEIN KELENJAR LUDAH Anopheles sp. TERINFEKSI P. bergheiPASCA IRADIASI GAMMA DENGAN TEKNIK SDS-PAGE UNTUK PENGEMBANGANVAKSIN MALARIA. Sporozoit merupakan tahapan siklus hidup parasit malaria yang palinginvasif dan merupakan kandidat vaksin paling tepat. Hasil penelitian menunjukkan bahwavaksin malaria yang dibuat dengan melemahkan sporozoit Plasmodium sp menggunakan sinargamma terbukti lebih efektif. Studi efek radiasi terhadap protein dalam pengembangan vaksiniradiasi juga berperan sangat penting. Tujuan dari penelitian ini adalah mengetahui profil proteinkelenjar ludah Anopheles sp terinfeksi sporozoit pasca iradiasi gamma dengan teknik Sodiumdodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Tahapan uji meliputipenginfeksian nyamuk Anopheles sp dengan P. berghei, pemeliharaan nyamuk terinfeksiselama 14-16 hari untuk memperoleh sporozoit, iradiasi nyamuk secara in vivo - in vitro,preparasi sampel kelenjar ludah dan elektroforesis pada SDS-PAGE 10% serta pewarnaanCommassie blue. Hasil uji menunjukkan perbedaan profill protein antara kelenjar ludahAnopheles sp terinfeksi dan tidak terinfeksi. Terdapat penambahan jumlah pita protein padadosis iradiasi lebih tinggi (200 Gy) dimana terdeteksi profil protein sporozoit P. berghei (BM 62kDa), tetapi tidak terdapat perbedaan profil circumsporozoite protein (CSP) antar dosis iradiasigamma 150, 175 dan 200 Gy. Hasil tersebut memberikan informasi dasar yang akan mengarahpada studi lanjut tentang peranan protein sporozoit dalam pengembangan vaksin malaria.Kata kunci : malaria, kelenjar ludah, P berghei, sporozoit, profil protein, sinar gamma
INDIKATOR BIOKIMIA SEL TERHADAP RADIASI PENGION Syaifudin, Mukh
Buletin Alara Vol 6, No 3 (2005): April 2005
Publisher : BATAN

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Radiasi pengion yang mengenai sistem hidup dapat menghasilkan serangkaian hasil akhir biologik meliputi proses pembengkakan, luka pada jaringan, karsinogenesis dan kematian. Tahap awal interaksi radiasi dengan materi biologi ini adalah pemindahan energi pada atom dan molekul yang menyebabkan pengionan dan eksitasi. Dosis letal sinar-X untuk pajanan akut seluruh tubuh pada manusia diperkirakan sebesar 4,5 Gy (450 rad). Dosis ini sangat kecil dan energi panas ekivalen dosis ini akan menambah suhu tubuh hanya sebesar 1/1000 oC. Daya mematikan dari energi sekecil ini berasal dari deposisi energi tak merata dalam sel dan melalui serangkaian proses biokimia yang menimbulkan luka. Proses tersebut meliputi pembentukan radikal bebas dan spesies aktif lain pada tingkat molekul dan pelepasan mediator biologik pada tingkat selular dan jaringan. Luka pada tingkat molekuler kemudian dapat menyebabkan luka pada jaringan yang dapat terjadi segera atau beberapa waktu sesudahnya. Proses lain dapat muncul melalui ketidakseimbangan hormonal dan pelepasan yang berlebihan agensia penyebab pembengkakan.
TINDAKAN PROTEKTIF TERHADAP KELENJAR TIROID PADA KECELAKAAN RADIASI Syaifudin, Mukh; Nazir, Fadil
Buletin Alara Vol 7, No 3 (2006): April 2006
Publisher : BATAN

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Kata-kata ”radiasi” dan ”nuklir” bagi orang awam adalah sesuatu yang menakutkan. Hal ini cukup beralasan karena nuklir pertama kali dikenali karena efek negatifnya. Hal ini juga tidak selamanya benar karena radiasi terbukti merupakan suatu alat dan teknologi yang telah dipergunakan secara meluas untuk membantu kebutuhan manusia seperti dalam hal kesehatan, makanan dan pertanian, industri, energi dan lingkungan. Banyak data dan informasi mengenai pemanfaatan teknologi berbasis radiasi ini, bahkan tidak dapat diperoleh atau dilakukan melalui teknologi lain, dan hasilnya pun dapat diandalkan (impresif). Sebagai contoh lebih dari 25 juta prosedur pencitraan nuklir dilakukan setiap tahunnya di seluruh dunia untuk diagnosa penyakit, dan setiap tahun hampir 10 juta orang memanfaatkan radiofarmasi (obat mengandung zat radioaktif) terutama untuk pengobatan kanker. Pada saat yang sama, teknik molekuler dan radioisotop saat ini sedang digunakan untuk mengembangkan teknologi dan pengukuran yang lebih efektif untuk melawan dan memberantas penyakit infeksi seperti malaria, hepatitis dan tuberkulosis. Teknologi berbasis radiasi yang telah terbukti efektif selama berpuluh-puluh tahun ini digunakan untuk memenuhi kebutuhan manusia pada hampir setiap sektor kehidupan.
Lack of Radioprotective Potential of Ginseng in Suppressing Micronuclei Frequency in Human Blood Lymphocyte under Gamma Irradiation LUSIYANTI, YANTI; ALATAS, ZUBAIDAH; SYAIFUDIN, MUKH
HAYATI Journal of Biosciences Vol 22, No 2 (2015): April 2015
Publisher : Bogor Agricultural University, Indonesia

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Ginseng appears to be a promising radioprotector for therapeutic or preventive protocols capable of attenuating the deleterious effects of radiation on human normal tissue. This research addresses results on the study of radioprotective potential of ginseng on radiation induced micronuclei in lymphocyte cells in vitro. The peripheral blood samples were exposed to gamma rays at doses of 0.0, 0.5, and 1.0 Gy and then added with 0, 100, and 1000 ug/mL of ginseng extract. These treated samples were cultured for micronuclei (MN) examination using standard procedure. The evaluation of incubation with ginseng extract for 24 h before irradiation was also done. Our results showed that there was no radioprotective effect of ginseng addition to the frequency of MN in lymphocyte cells. Pre-incubation with ginseng extract before irradiation also did not effectively suppress the MN frequency. This research lacks to prove the ginseng’s radioprotective potential that maybe related to its immunomodulating capabilities and its capabilities in scavenging free radicals induced by radiation or in attenuating the deleterious effects of radiation and its important role in increasing levels of several cytokines.
Analysis of the resistance of M. tuberculosis to fluoroquinolon and the implementation of nuclear based biomolecular technique. Syaifudin, Mukh; Septiani, Dewi
INDONESIAN JOURNAL OF PHARMACY Vol 22 No 2, 2011
Publisher : Faculty of Pharmacy Universitas Gadjah Mada, Yogyakarta, Skip Utara, 55281, Indonesia

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Tuberculosis (TB) is still a problem in community health with high rate of mortality.  The  case  became  much  more  complicated  due  to  emerge  of Mycobacterium  tuberculosis which  are  resistant  to  the  drugs.  This  caused  the movement  of  attention  from  the  first  line  drugs  to  fluoro-quinolon  (FQ)  as alternative drug. The aim of this research was to do analysis the mutation which causing  the  resistance  of  bacterial  through  nucleic acid  alterations  with polymerase  chain  reaction  (PCR)  and  single  strand  conformation  polymorphism (SSCP)  technique.  Analysis  was  done  on  gyrA  and  gyrB  genes  encoding  DNA gyrase of bacterial and closely related to FQ resistance in 100 of sputa samples of  positive  BTA  test  results.  DNA  of  M.  tuberculosis strain  H37Rv  was  used  as control. From analysis on gyrA gene it was known that 57 samples were positive PCR  and  no  resistant  sample  was  found.  For  gyrB  gene,  only  12  of  them  were positive  PCR  and  again  there  was  no  samples  had  mutation  as  cause  of resistance.  These  mean  that  FQ  could  be  used  as  replacement  drug.  Molecular detection  technique  was  known  fast  and  specific  for assessing  bacterial resistance.  Researcher  proves  that  searching  for  P32-radioisotope  labeled  DNA alteration  was  more  sensitive.  Hopefully  this  results  of  experiment  can  be implemented  in  medication  with  more  effective  and  support  diagnose  results so that it will lowering the risk of patient mortality.Key words : M. tuberculosis, fluoro-quinolon, resistance, PCR, SSCP
Perubahan Molekular Gen Penekan Tumor P-53 Akibat Pajanan Radiasi Pengion Syaifudin, Mukh
Jurnal Forum Nuklir JFN Vol 6 No 1 Mei 2012
Publisher : BATAN

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