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All Journal HAYATI Journal of Biosciences Microbiology Indonesia Health Science Journal of Indonesia Indonesian Journal of Cancer The Indonesian Biomedical Journal Molecular and Cellular Biomedical Sciences (MCBS) Acta Biochimica Indonesiana Jurnal Ilmu Kefarmasian Indonesia The New Ropanasuri Journal of Surgery Muhammadiyah Journal of Geriatric Indonesian Journal of Cancer eJournal Kedokteran Indonesia Makara Journal of Science Molekul: Jurnal Ilmiah Kimia
Mohamad Sadikin
Department Of Biochemistry And Molecular Biology, Faculty Of Medicine, Universitas Indonesia, Jakarta
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemistry Dentistry Earth & Planetary Sciences Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Nursing Public Health

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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.
Intermittent Exposure to Hypobaric Hypoxia Increases VEGF, HIF-1α, and Nrf-2 Expressions in Brain Tissue Wardaya, Wardaya; Sukmawati, Dewi; Ibrahim, Nurhadi; Ferdinal, Frans; Mudjihartini, Ninik; Sadikin, Mohamad; Jusman, Sri Widia A.; Satriotomo, Irawan; Mulyawan, Wawan
The Indonesian Biomedical Journal Vol 17, No 2 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i2.3519

Abstract

BACKGROUND: Hypoxia inducible factor-1α (HIF-1α), nuclear factor erythroid 2-related factor 2 (Nrf-2), and vascular endothelial growth factor (VEGF), play a crucial role as neuroprotective factors. Currently, there is a lack of studies examining the biomolecular responses of the brain to intermittent hypoxia resulting from various pressures. This study was conducted to investigate the physiological responses, histopathological features, and cellular adaptive responses in the brains of rats that were intermittently exposed to hypobaric hypoxic conditions.METHODS: Thirty male Sprague Dawley rats were divided into six groups: a control group and five treatment groups exposed to hypobaric hypoxia. The treatment groups were placed in a hypobaric chamber simulating an altitude of 3,048 meters for 1 hour/day for 1, 7, 14, 21, and 28 days. After exposure, brain tissue was collected for histopathological analysis and protein quantification of HIF-1α, Nrf-2, cytoglobin (Cygb), neuroglobin (Ngb), VEGF, malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT).RESULTS: In the brain, intermittent hypobaric hypoxia significantly increased HIF-1α expression (p=0.000) and its downstream proteins Cygb (p=0.000), and VEGF (p=0.001), with a peak at 14x IHH exposure compared to control. This was followed by a significant increase in Nrf-2 expression (p=0.000), SOD (p=0.000), Gpx (p=0.000), and CAT activity (p=0.000), indicating an adaptive antioxidant response. Conversely, MDA levels was decreased with prolonged exposure, suggesting reduced oxidative damage.CONCLUSION: IHH elevates HIF-1α, Nrf-2, and oxidative stress markers, triggering an adaptive antioxidant response in the rat’s brains.KEYWORDS: HIF-1α, intermittent hypobaric hypoxia, Nrf-2, oxidative stress
Lower IGF-1 and Alkaline Phosphatase Activity Blood Levels in Stunted Children with Soil-Transmitted Helminth Infections Lio, Tiara Mayang Pratiwi; Wibowo, Heri; Sadikin, Mohamad; Jusman, Sri Widia A.
The Indonesian Biomedical Journal Vol 17, No 2 (2025)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v17i2.3454

Abstract

BACKGROUND: Stunting due to malnutrition and soil-transmitted helminth (STH) chronic infections can cause disturbances in bone formation processes during growth. Reduced levels of insulin-like growth factor 1 (IGF-1) lead to reduced osteoblast activity which can be monitored through alkaline phosphatase (ALP) and osteocalcin (OCN). Unfortunately, studies that assessed IGF-1, OCN, and ALP together in stunted children with STH infection are still limited. Therefore, in this study, the osteoblast activity was monitored by measuring IGF-1, OCN, and ALP in stunted and normo-stature children with STH infection.METHODS: A case-control study involving 28 stunted and 33 normal-stature children was conducted. Blood plasma IGF-1 and OCN levels were measured using the enzyme linked immunosorbent assay (ELISA) method, while ALP activity was measured using the colorimetric method. STH infection was examined by direct smear method. The parameters were analyzed and compared between the stunted and normo-stature children.RESULTS: Although in stunted boys there was no significant difference, however in stunted girls with positive STH infection, a decrease in IGF-1 (<36.60 ng/mL vs. ≥36.60 ng/mL) showed an impact on the decrease of OCN levels (54.68 (42.22-144.54) ng/mL vs. 104.55 (86.14-392.73) ng/mL; p=0.047). ALP activity in children with positive STH infection showed that ALP activity in stunted children was lower than in normo-stature children (18 (10-671) U/L vs. 228.50 (16-574) U/L; p=0.005).CONCLUSION: The blood levels of IGF-1 and ALP activity in stunted and positive STH infected children tend to be lower than in normo-stature children.KEYWORDS: stunted, normo-stature, IGF-1, bone alkaline phosphatase, osteocalcin
Isolation and Purification of Breast Milk Folate Binding Protein: Salting-Out and Chromatography Techniques Saleh, Mgs. M. Irsan; Subandrate, Subandrate; Gunarti, Dwirini Retno; Hermansyah, Hermansyah; Hafy, Zen; Kesuma, Yudianita; Sadikin, Mohamad
Molekul Vol 20 No 1 (2025)
Publisher : Universitas Jenderal Soedirman

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20884/1.jm.2025.20.1.11303

Abstract

ABSTRACT. Folate binding protein (FBP) is a protein in breast milk that plays a role in the regulation and bioavailability of folic acid. In contrast to cow's milk FBP, information about breast milk FBP is still limited. This research aims to determine the isolation and purification methods of breast milk FBP and the molecular weight of breast milk FBP. The sample in this study was 1000 mL of breast milk. Breast milk was prepared in several stages to yield whey. Isolation and purification of FBP from whey were carried out in stages, salting-out, ion exchange chromatography, and affinity chromatography. Whey salting-out with 95% saturation of ammonium sulfate could precipitate folate-binding proteins. This precipitate showed three peaks on DEAE chromatography. Peak II DEAE 95% was thought to be a negatively charged folate-binding protein. Peak II DEAE 95% also showed the presence of two peaks on affinity chromatography. It was believed that Peak II AF 95% was a pure folate-binding protein. Peak II AF 95% showed the presence of a single band on SDS-PAGE and western blot. This indicated that the folate-binding protein was 100% pure. FBP can be isolated from breast milk by the salting-out method using 95% ammonium sulfate, DEAE chromatography, and affinity chromatography. FBP from breast milk has a molecular weight of approximately 37 kDa. The final level of FBP isolated from breast milk is approximately 0.022 mg/mL. The successful isolation of FBP from breast milk provides an opportunity to use it to understand the clinical role of FBP in increasing folic acid levels in both breast milk and infant serum, as well as to develop methods for determining folic acid levels in these fluids. Keywords: Breast milk, folate binding protein, isolation, purification, molecular weight
Purification of total IgG from sars-cov-2 convalescent serum Lusinanto, Arfat; Gantini, Ria Syafitri Evi; Gunarti, Dwirini Retno; Sadikin, Mohamad
Acta Biochimica Indonesiana Vol. 8 No. 2 (2025): Acta Biochimica Indonesiana
Publisher : Indonesian Society for Biochemistry and Molecular Biology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32889/actabioina.203

Abstract

Background: Convalescent plasma contains neutralizing antibodies against SARS-CoV-2 but also potentially harmful inflammatory cytokines. Purified immunoglobulin G fractions offer a safer therapeutic alternative by concentrating antibodies while removing inflammatory mediators. Objective: To establish a systematic protocol for purifying total IgG from SARS-CoV-2 convalescent serum using sequential chromatographic techniques. Methods: Serum from 90 recovered donors was pooled into three independent samples. Purification employed four sequential steps: ammonium sulfate precipitation (50% saturation), Sephadex G-100 size-exclusion chromatography, DEAE-Cellulose ion-exchange chromatography, and Protein A affinity chromatography. Purity was assessed by native polyacrylamide gel electrophoresis and radial immunodiffusion. Results: Starting from serum with 19.68 ± 7.27 mg/mL IgG and 110.47 ± 11.99 mg/mL total protein, the four-step purification yielded final IgG concentration of 1.14 ± 0.70 mg/mL with 1.19 ± 0.16 mg/mL total protein. This achieved 6.3-fold purification with purity ratio of 1.01 ± 0.38 and 5.8% recovery. Native PAGE confirmed high purity with a single IgG band. Conclusion: Sequential chromatography successfully purified total IgG from convalescent serum, providing a laboratory-scale method for preparing safer immunoglobulin therapeutics.
Detection of latent tuberculosis infection in household contacts of drug-resistant tuberculosis patients using interferon-gamma release assay: a study at Universitas Indonesia Hospital Indratmo, Muhammad Faris; Handayani, Diah; Kusumaningrum, Ardiana; Iswanti, Febriana Catur; Sadikin, Mohamad
Acta Biochimica Indonesiana Vol. 8 No. 2 (2025): Acta Biochimica Indonesiana
Publisher : Indonesian Society for Biochemistry and Molecular Biology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.32889/actabioina.214

Abstract

Background: Drug-resistant tuberculosis (DR-TB) poses significant public health challenges in Indonesia. Household contacts of DR-TB patients face elevated risk of Mycobacterium tuberculosis infection, which may remain latent and asymptomatic. Objective: This study aimed to assess the prevalence of latent tuberculosis infection (LTBI) among household contacts of DR-TB patients using interferon-gamma release assay (IGRA). Methods: This cross-sectional study was conducted at Universitas Indonesia Hospital from February to May 2023. Eighteen asymptomatic household contacts from six confirmed DR-TB index cases were enrolled. Participants underwent clinical evaluation, chest radiography, and LTBI screening using the QuantiFERON-TB Gold Plus (QFT-Plus) assay. Results: Among 18 participants (mean age 33.3 years; 55.6% female), 8 (44.4%) tested positive for LTBI, while 10 (55.6%) tested negative. The highest IGRA positivity rates were observed in adolescents aged 12–16 years (66.7%) and young adults aged 17–25 years (60.0%). All participants were clinically asymptomatic with normal chest radiographs. Conclusion: This study demonstrates substantial LTBI prevalence among household contacts of DR-TB patients. The findings underscore the importance of systematic contact tracing, IGRA-based screening, and timely tuberculosis preventive therapy to reduce disease transmission and progression in high-risk populations.
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 Ardiana Kusumaningrum 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 Handayani 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 Gantini, Ria Syafitri Evi HANIFAH RAHMI Hans -Joachim Freisleben Hans Joachim Freisleben HANS-JOACHIM FREISLEBEN Heri Wibowo Hermansyah Hermansyah Indra G. Mansur Indratmo, Muhammad Faris Jeanne A. Pawitan JEANNE ADIWINATA PAWITAN Juniarti . Jusman, Sri Widia Azraki Lailan Safina Nasution Lio, Tiara Mayang Pratiwi Lusinanto, Arfat 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