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Expression of Simian Retrovirus Type D Serotype 2 Envelope in Insect Cell Using Baculovirus Expression Vector System DIAH ISKANDRIATI; MOHAMAD SADIKIN; JOKO PAMUNGKAS
Microbiology Indonesia Vol. 3 No. 2 (2009): August 2009
Publisher : Indonesian Society for microbiology

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (167.528 KB) | DOI: 10.5454/mi.3.2.8

Abstract

Simian retrovirus type-D (SRV) is a causative agent of simian acquired immunodeficiency syndrome in Asian macaques, and can serve as a viral model in understanding of retrovirus infection because of some similarities to human AIDS pathogenesis. Study of infection and pathogenesis of SRV in macaques could be a strategy of vaccine and antiviral development for preventive and therapeutic purposes. We expressed the SRV-2 envelope gene using baculovirus expression vector system and transfected it to Spodoptera frugiferda insect cell line for SRV-2 recombinant protein production. Analysis using PCR and sequencing technique of recombinant in the passage-3 viral stock indicated the occurrence of recombination between SRV-2 envelope and baculovirus genome. Purification using immobilized metal ion affinity chromatography Ni2+-NTA to recombinant protein could minimize the presence non-specific proteins. The SDS-PAGE analysis showed a specific protein for SRV-2 gp70 envelope. Western blot analysis of this purified protein indicated a specific reaction with anti-SRV-2 antibody positive of Macaca fascicularis serum shown as SRV-2 gp70 envelope band.
HDAC2 and PCNA expression is correlated to decreasing of endoxifen sensitivity in human breast cancer stem cells ALDH+ Syarifah Dewi; Mohamad Sadikin; Muchlis Ramli; Septelia Inawati Wanandi
Health Science Journal of Indonesia Vol 10 No 2 (2019)
Publisher : Sekretariat Badan Penelitian dan Pengembangan Kesehatan

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22435/hsji.v12i2.2449

Abstract

Latar belakang: Sel punca kanker payudara (breast cancer stem cells/BCSC) adalah subpopulasi sel kanker yang memiliki kemampuan menghasilkan tumor baru dan bersifat seperti sel punca. Penelitian kami sebelumnya menggunakan jaringan kanker payudara mengungkapkan bahwa ekspresi gen histone deacetylase 2 (HDAC2) dan proliferating cell nuclear antigen (PCNA) ditemukan perbedaan signifikan setelah terapi neoajuvan hormon dan kemoterapi. Penelitian ini bertujuan untuk menganalisis hubungan antara ekspresi HDAC2 dan PCNA dengan kelangsungan hidup sel punca kanker payudara dengan penanda aldehyde dehydrogenase + (ALDH+) yang diberi perlakuan endoksifen. Metode: Sampel adalah BCSC primer manusia ALDH+ yang diberi perlakuan endoksifen 4 uM masingmasing selama 2, 4, 6, 8, 10, 12, 14 hari. Viabilitas sel dilihat dengan menggunakan trypan blue dan ekspresi mRNA HDAC2 dan PCNA ditentukan menggunakan qRT-PCR. Hasil: Viabilitas BCSCs ALDH + menurun setelah 2 sampai 4 hari pemberian endoksifen. Pada periode ini juga didapatkan ekspresi mRNA HDAC2 dan PCNA mengalami penurunan. Tetapi setelah pemberian endoksifen selama 8 hari, viabilitas BCSCs ALDH + mengalami peningkatan dan ditemukan peningkatan yang signifikan pada hari ke-14 pemberian endoksifen. Ekspresi mRNA HDAC2 dan PCNA juga menunjukkan peningkatan mulai pada hari ke-8 dan terus meningkat hingga hari ke-14 pemberian endoksifen. Penelitian ini menunjukkan pola yang sama antara ekspresi mRNA HDAC2 dan PCNA dan viabilitas sel. Kesimpulan: Induksi endoksifen yang lama menurunkan sensitivitas efek endoksifen pada BCSC manusia dan ekspresi HDAC2 dan PCNA berkorelasi dengan viabilitas BCSC manusia setelah induksi endoksifen. (Health Science Journal of Indonesia 2019;10(2):77-81) Kata kunci: sel punca kanker payudara, viabilitas sel, HDAC2, PCNA, endoksifen Abstract Background: Breast cancer stem cells (BCSCs) are subpopulation of cancer cells that has the ability to generate new tumor and similar properties to stem cell. Our previous study using breast cancer patients revealed that gene expression of histone deacetylase 2 (HDAC2) and proliferating cell nuclear antigen (PCNA) were significantly altered after neoadjuvant hormone and chemotherapy. This study aimed to analyze the correlation between HDAC2 and PCNA expressions with the viability of breast cancer stem cells aldehyde dehydrogenase + (BCSC ALDH+) treated by endoxifen. Method: Samples are human primary BCSCs ALDH+ that treated with 4 uM of endoxifen for 2, 4, 6, 8, 10, 12, 14 days, respectively. Cell viability was measured using trypan blue exclusion assay and the mRNA expressions of HDAC2 and PCNA were determined using qRT-PCR. Results: The viability of BCSCs ALDH+ was decreased after 2 days until 4 days-endoxifen treatment. It also demonstrated that mRNA expression of HDAC2 and PCNA were decreased in this period. But after 8 days endoxifen treatment, the viability of BCSCs ALDH+ was increased. The increasing of viability was higher in 14 days-endoxifen treatment. The mRNA expression of HDAC2 and PCNA also showed increasing begin on 8 days and continued to increase until 14-days endoxifen treatment. We found a similar pattern between HDAC2 and PCNA expression and cell viability. Conclusion: Prolonge endoxifen treatment decrease sensitivity of endoxifen effect in human BCSC and the expression of HDAC2 and PCNA are correlated to human BCSCs viability after endoxifen treatment. (Health Science Journal of Indonesia 2019;10(2):77-81) Keywords: human breast cancer stem cells, viability, HDAC2, PCNA, endoxifen
Peningkatan Kadar Imunosupresan IDO (Indoleamin 2,3-dioksigenase) pada Supernatan Kultur Sel Punca Mesenkim yang Distimulasi dengan Agregat Imunoglobulin G DIAN RATIH LAKSMITAWATI; JEANNE ADIWINATA PAWITAN; MOHAMAD SADIKIN; CAROLINE TAN SARDJONO
JURNAL ILMU KEFARMASIAN INDONESIA Vol 12 No 1 (2014): JIFI
Publisher : Fakultas Farmasi Universitas Pancasila

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (480.939 KB)

Abstract

Mesenchymal stem cells have been known to have the nature of supressing immune response. This cell have been proven to reduce the symptoms of organ transplants rejection. Clinically proofing is also being performed on an autoimmune disease caused by the complex immune such as systemic lupus erythematosus (SLE). This study aimed to know aggregated IgG-stimulating effects of mesenchymal stem cells in relation to IDO secretion, an immunosuppressant agent. Mesenchymal stem cells was isolated from liposuction waste and characterized based on its surface molecule accrording to ISCT criterias. The confirmed cell culture was used to the next culture by the addition of heat aggragated immunoglobulins gamma (HAGG) for six days. IDO secreted on supernatant culture was collected and assayed by elisa method at 450 nm. The results showed that there was elevated levels of IDO (16.73-49.5%) on culture with HAGG stimulation than without stimulation. This study could be a part of the basic
Isolasi dan Karakterisasi Protease Serupa Tripsin (PST) dari Lactobacillus plantarum FNCC 0270 TRISMILAH TRISMILAH; WAHONO SUMARYONO; AMARILA MALIK; MOHAMAD SADIKIN
JURNAL ILMU KEFARMASIAN INDONESIA Vol 12 No 1 (2014): JIFI
Publisher : Fakultas Farmasi Universitas Pancasila

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (665.7 KB)

Abstract

Trypsin is an enzyme which is important for research as well as for the pharmaceutical industry, medicine and health, which is especially developed as a raw material for digestive enzymes. This study aimed to characterize PST from Lactobacillus plantarum FNCC 0270 that have not been studied as a producer of trypsin. Fermentation for producing PST by L . plantarum FNCC 0270 was performed in the LKB fermentor with working volume of 3.5 liters, at temperature of 37 oC, pH value of 8, agitation rate of 77 rpm and aeration of 0.5 vvm. The media composition were 3.64% baker’s yeast, 1.21% glucose, 0.13% skim milk. The fementation resulted in enzyme activities of 0.00129 U/mL and protein content of 0.49 mg/mL. Purification of the PST conducted by ultrafiltration Hollow Fiber Catridge 5 kD, presipitated with ammonium sulfate saturated (30-70%), 30 kD membrane polyesthersulfone, ion exchange column chromatography Q-XL and HiTrap affinity column chromatography. All of system increased purity of the crude PST. SDS-PAGE of PST indicated the molecular weights of 47.35; 38.42;21.40 and 12.96 kD. The PST was stable at pH value of 8 and temperature range of 25-35 oC. It is proved that the same half-life was 693.2 minutes. Km and Vmax values of the PST were 0.231 mM and 0.001 U/mL, respectively using N-α-benzoyl-DL-arginine-p-nitroanilide as the substrate. PST was inhibited by EDTA, Ca2+, Zn2+, Mg2+, Mn2+ and specific substrates. Based on the result of characterization of PST from L. plantarum FNCC 0270 can be concluded that the trypsin–like protease (PST) was trypsin.
Ekspresi Relatif mRNA Hypoxia Inducible Factor-1? pada Sel Glioma Penderita Novi Silvia Hardiany; Mohamad Sadikin; Septelia Inawati Wanandi
Indonesian Journal of Cancer Vol 7, No 1 (2013): Jan - Mar 2013
Publisher : National Cancer Center - Dharmais Cancer Hospital

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (530.441 KB) | DOI: 10.14414/ijoc.v7i1.272

Abstract

Glioma merupakan tumor otak primer yang cukup sering ditemukan di Indonesia di antara keganasan otak lainnya. Penderita glioma mempunyai angka harapan hidup yang rendah dikarenakan seringnya mengalami resistansi terapi. Hipoksia merupakan salah satu faktor yang mempengaruhi respons terapi radiasi. Penelitian ini menganalisis ekpresi mRNA HIF-1?, sebagai petanda jaringan hipoksia, pada glioma derajat rendah dibandingkan dengan glioma derajat tinggi. Sampel terdiri dari 15 jaringan glioma derajat rendah, 6 jaringan glioma derajat tinggi, dan 21 sel lekosit dari penderita glioma sebagai kontrol sel normal. Ekspresi relatif mRNA HIF-1? ditentukan secara kuantitatif dengan menggunakan Real Time RT-PCR. Hasil penelitian menunjukkan bahwa ekspresi mRNA HIF-1? pada glioma derajat tinggi lebih tinggi dibandingkan dengan glioma derajat rendah, namun tidak bermakna secara statistik. Hal yang menarik yaitu ditemukan peningkatan ekspresi mRNA HIF-1? dengan nilai yang bervariasi pada 73 % sampel glioma derajat rendah dibandingkan dengan sel lekosit sebagai kontrol sel normal. Begitu juga pada glioma derajat tinggi ditemukan peningkatan mRNA HIF-1? dengan nilai yang bervariasi pada 100% sampel. Dengan demikian, dapat disimpulkan bahwa terdapat variasi derajat hipoksia pada glioma derajat rendah dan derajat tinggi yang mungkin mempengaruhi keberhasilan terapi radiasi.Kata kunci: Glioma, mRNA HIF-1?, hipoksia
Mitochondrial Adaptations of Obese Adult Rat Visceral Adipocytes which Differ in Nutritional Status during Childhood Lailan Safina Nasution; Ahmad Aulia; Sri Widia A. Jusman; Mohamad Sadikin
eJournal Kedokteran Indonesia Vol 9, No. 3 - Desember 2021
Publisher : Faculty of Medicine Universitas Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (243.296 KB) | DOI: 10.23886/ejki.9.48.197-202

Abstract

Mitochondria are highly dynamic organelles that can adapt to different environmental stimuli. This study aims to analyze differences in the function and biogenesis of mitochondria and anaerobic glycolysis activity between obese adult rats that were undernourished in childhood (LCD28) and those which were normal (SD28) or already fat (HFD28). Sprague-dawley rats were assigned groups and given diet treatments from December 2017 until August 2018 in the Laboratory Animal Management Unit, Faculty of Veterinary Medicine, Bogor Agricultural Institute. To observe mitochondrial function and biogenesis, MnSOD activity and PGC1α levels were analyzed using ELISA. To investigate the existence of anaerobic glycolysis, LDH activity was analyzed using a spectrophotometry method. The results showed that MnSOD activity and PGC1α levels of LCD28 increased compared to the other groups, signifying higher mitochondrial function and biogenesis. Meanwhile, no significant difference in LDH activity was found, signifying similar levels of anaerobic glycolysis.
Optimization of Trypsin-like Protease Production by Lactobacillus plantarum FNCC 0270 using Response Surface Methodology Trismilah,; Nurhasanah, Astutiati; Sumaryono, Wahono; Malik, Amarila; Sadikin, Mohamad
Makara Journal of Science Vol. 19, No. 2
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

The purpose of this study was to get optimum medium composition and agitation to trypsin-like protease production by Lactobacillus plantarum FNCC 0270. The medium composition and agitation for enzyme production was optimized using Central Composite Design and Response Surface Method with Design Expert software version 7.1.5 Fermentation was carried out in erlenmeyer flasks at initial pH 8, 37 °C, using an incubator shaker at 87.5 rpm. The best results showed an enzyme activity of 1.0 mU/mL, a protein level of 0.557 mg/mL, and desirability value of 0.740. Numerical optimization was performed to approach the ideal state of the fermentation or the desirability value of 1. The medium composition containing of 3.64% baker's yeast, 1.21% glucose, and 0.13% skim milk was used for the fermentation. The enzyme activity of 1.51 mU/mL and protein level of 0.205 mg/mL can be achieved. After numerical optimization, the fermentation process was verified in erlenmeyer flasks with incubator shaking at 77 rpm, initial pH 8, 37 °C, and 15 h fermentation. The verification results showed that the enzyme activity of 1.273 ± 0.227 mU/mL and protein level of 0.248 ± 0.012 mg/mL.
The Role of Hypoxia-inducible Factor in Mycobacterium tuberculosis-infected Macrophages Fitriana, Nina; Iswanti, Febriana Catur; Sadikin, Mohamad
Molecular and Cellular Biomedical Sciences Vol 8, No 1 (2024)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v8i1.405

Abstract

Tuberculosis is caused by Mycobacterium tuberculosis infection. During M. tuberculosis infection, there is a decrease in the partial pressure of oxygen in the granuloma microenvironment, which causes the hypoxia-inducible factor (HIF) to become stable. HIF functions as a transcription factor that regulates the expression of genes crucial for metabolic adaptation in hypoxic conditions. Recent research suggests that HIF plays a vital role in infectious and inflammatory conditions. Several studies have demonstrated that HIF signaling can enhance macrophages antimicrobial activity and bactericidal effect against M. tuberculosis, such as increasing macrophage autophagy, enhancing the effects of rifampicin, inhibiting p38 MAPK signaling, enhancing the regulation of effector antimicrobial pathways mediated by human β defensin 2 (hBD2) and vitamin D receptor (VDR), redirecting energy metabolism to glycolysis, and producing various cytokines. All these responses ultimately result in the inhibition of intracellular M. tuberculosis growth. HIF has therapeutic implications, potentially being a new candidate for host-directed therapy as a complement to existing antituberculosis drugs. Understanding the role of HIF in macrophages during M. tuberculosis infection and comprehending the host-pathogen relationship with M. tuberculosis is advantageous for developing future therapies.Keywords: Mycobacterium tuberculosis, macrophages, hypoxia-inducible factor
Isolation and Purification of Thiamine Binding Protein from Mung Bean GUNARTI, DWIRINI RETNO; RAHMI, HANIFAH; SADIKIN, MOHAMAD
HAYATI Journal of Biosciences Vol. 20 No. 1 (2013): March 2013
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (78.56 KB) | DOI: 10.4308/hjb.20.1.1

Abstract

Thiamine has fundamental role in energy metabolism. The organs mostly sensitive to the lack of thiamine levels in the body are the nervous system and the heart. Thiamine deficiency causes symptoms of polyneuritis and cardiovascular diseases. Because of its importance in the metabolism of carbohydrates, we need to measure the levels of thiamine in the body fluids by using an easy and inexpensive way without compromising the sensitivity and selectivity. An option to it is thiamine measurement based on the principle of which is analogous to ELISA, in which a thiamine binding protein (TBP) act by replacing antibodies. The presence of TBP in several seeds have been reported by previous researchers, but the presence of TBP in mung beans has not been studied. This study was aimed to isolate and purify TBP from mung bean. The protein was isolated from mung bean  through salting out by ammonium sulphate of 40, 70, and 90% (w/v). TBP has a negative charge as shown by cellulose acetate electrophoresis. The result obtained after salting out by ammonium sulphate was further purified bymeans of DEAE-cellulose chromatography and affinity chromatography. In precipitation of 90% of salting out method, one peak protein was obtained by using affinity chromatography. The protein was analyzed by SDS PAGE electrophoresis. The result of SDS PAGE electrophoresis showed that TBP has a molecular weight of 72.63 kDa.
Kadar Neuroglobin dan Sitoglobin dalam Plasma, Cairan Serebro Spinalis, dan Jaringan Otak Pasien Strok Hemoragik Mudjihartini, Ninik; Saekhu, Mohamad; Jusman, Sri Widia A.; Sadikin, Mohamad
Muhammadiyah Journal of Geriatric Vol 3, No 1 (2022): Muhammadiyah Journal of Geriatric
Publisher : Faculty of Medicine and Health Universitas Muhammadiyah Jakarta

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24853/mujg.3.1.1-8

Abstract

Latar belakang: Otak memerlukan oksigen yang banyak selain glukosa. Hipoksia iskemik karena strok hemoragik atau Spontaneous intracerebral hemorrhage (sICH) dapat mengganggu suplai oksigen dan nutrisi ke otak berakibat produksi energi di otak akan menurun. Deplesi energi ini menyebabkan kerusakan dan kematian sel otak terjadi lebih cepat. Neuroglobin (Ngb) dan sitoglobin (Cygb) merupakan protein golongan globin yang terdapat di otak dan berperan sebagai protein pengikat oksigen di mitokondria. Tujuan: Penelitian ini bertujuan mendapatkan gambaran/profil kadar Ngb dan Cygb, di plasma, cairan serebro spinal (CSS), dan jaringan otak pasien strok hemoragik. Metode: Penelitian ini merupakan penelitian lanjutan menggunakan sampel plasma, CSS, dan jaringan otak yang diperoleh saat kraniotomi evakuasi hematoma pasien strok hemoragik sICH di rumah sakit Cipto Mangunkusumo dan rumah sakit lainnya di Jakarta. Kadar protein Ngb dan Cygb dari plasma, CSS, dan jaringan otak diukur dengan metode ELISA. Hasil: Rerata kadar Ngb otak adalah 0,058 ng/mg protein otak, sedangkan di plasma dan CSS masing-masing adalah 0,017 ng/mg protein otak dan 0,013 ng/mg protein otak atau 29,31% dan 22,41% dari rerata kadar Ngb otak. Rerata kadar Cygb otak adalah 4,943 ng/mg protein otak, di CSS adalah 1,685 ng/mg protein otak, atau 25,26% dari rerata Cygb otak, sedangkan di dalam plasma hampir tidak terdeteksi. Simpulan: Pada keadaan hipoksia oleh karena strok hemoragik sICH, protein Ngb dan Cygb dapat diukur di plasma, CSS, dan jaringan otaknya.
Co-Authors . Rusdi Ahmad A. Jusuf Ahmad Aulia Ahmad Aulia Jusuf Ahmad R. Utomo AMARILA MALIK Amarila Malik Andi N.K. Syarifin, Andi N.K. ANDREW ROBERT COSSINS Angelina Stevany Regina Masengi Ani R. Prijanti Arief Budi Witarto Aryenti . Asri Rasad ASTUTIATI NURHASANAH Astutiati Nurhasanah Bethy S. Hernowo Caroline T. Sardjono CAROLINE TAN SARDJONO Cicia Firakania, Cicia DARYL ROBERT WILLIAMS Dewi Sukmawati Diah Iskandriati Dian R. Laksmitawati Dian Ratih Laksmitawati Dwirini Retno Gunarti Ekawati, Maria Erni Hernawati Purwaningsih, Erni Hernawati Fanny Septiani Farhan Febriana C. Iswanti Febriana Catur Iswanti Franciscus D. Suyatna Frans Ferdinal HANIFAH RAHMI Hans -Joachim Freisleben Hans Joachim Freisleben HANS-JOACHIM FREISLEBEN Heri Wibowo Hermansyah Hermansyah Indra G. Mansur Jeanne A. Pawitan JEANNE ADIWINATA PAWITAN Juniarti . Jusman, Sri Widia Azraki Lailan Safina Nasution Lio, Tiara Mayang Pratiwi Mohamad Saekhu Muchlis Ramli Nani Dharmasetiawani Nina Fitriana, Nina Ninik Mudjihartini Noorwati Sutandyo Novi S. Hardiany Novi Silvia Hardiany Nurhadi Ibrahim Nurjati C. Siregar Oentoeng Soeradi Pamungkas, Joko Praditi, Citra Radiana D. Antarianto Rahmawati Ridwan Reni Paramita Rini Puspitaningrum RONDANG ROEMIATI SOEGIANTO Rostika Flora Saleh, Mgs. M. Irsan Samsuridjal Djauzi Sari, Dewi H. Sarsanti, Pungguri Ayu Nega Satriotomo, Irawan Septelia I. Wanandi SEPTELIA INAWATI WANANDI SEPTELIA INAWATI WANANDI Siregar, Nurjati Siti Rahmawati Achyat Siufui Hendrawan Sri S. Ningsih, Sri S. Sri W.A. Jusman Sri Widia A.Jusman Subandrate Syarifah Dewi Tiwuk Susantiningsih Tomohiko Yamazaki Trismilah Trismilah TRISMILAH TRISMILAH Trismilah, Uly A. Nikmah, Uly A. Wahono Sumaryono Wardaya Wardaya Wawan Mulyawan Wawan Mulyawan Yefta Moenadjat Yudianita Kesuma, Yudianita Yuhernita . Zen Hafy