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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.
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: Although convalescent plasma contains neutralizing anti-SARS-CoV-2 antibodies, co-existing inflammatory mediators pose safety risks in critically ill recipients. Purified IgG preparations offer a safer alternative by concentrating therapeutic antibodies while eliminating these harmful components. Objective: To establish a systematic protocol for purifying total IgG from SARS-CoV-2 convalescent serum using sequential chromatographic techniques. Methods: Serum from 90 RT-PCR-confirmed 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 and identity of IgG fractions were assessed by native polyacrylamide gel electrophoresis and radial immunodiffusion. Results: Starting from serum containing 19.68 ± 7.27 mg/mL IgG and 110.47 ± 11.99 mg/mL total protein, the four-step purification yielded a final IgG concentration of 1.14 ± 0.70 mg/mL with total protein of 1.19 ± 0.16 mg/mL, representing 6.3-fold purification with a final IgG-to-total protein purity ratio of 1.01 ± 0.38 and an overall recovery of 5.8%. Native PAGE confirmed high purity with a single dominant IgG band. Conclusion: Sequential chromatography yielded near-homogeneous IgG from SARS-CoV-2 convalescent serum, offering a laboratory-scale approach 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.
The Effect of Triglycerides and Total Cholesterol Levels of Lipemic Plasma on The Results of HBsAg Screening Using Chemiluminescence Immunoassay Astriani , Ranti Dwi; Ritchie, Ni Ken; Sadikin, Mohamad
JURNAL INFO KESEHATAN Vol 23 No 4 (2025): JURNAL INFO KESEHATAN
Publisher : Research and Community Service Unit, Poltekkes Kemenkes Kupang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.31965/infokes.Vol23.Iss4.1809

Abstract

The results of the Immunochromatographic Method for Infectious Disease Testing (IMLTD) may vary depending on the presence of lipids in donor blood samples, potentially leading to false‐negative or false‐positive outcomes in hepatitis B surface antigen (HBsAg) screening. This study aimed to evaluate the effect of lipid interference on HBsAg screening using the chemiluminescent immunoassay (CLIA) method, to determine the range of triglyceride levels that may influence test results (100 mg/dL to >3000 mg/dL), and to assess practical lipid removal methods to ensure blood safety. An experimental laboratory study was conducted using two sample groups. The first group consisted of six bags of non-lipemic, HBsAg-reactive plasma that were rendered lipemic by the addition of Lipofundin. The second group included 25 bags of lipemic donor plasma that were non-reactive in the initial IMLTD screening and subsequently subjected to lipid removal treatments. Lipid reduction was performed using centrifugation, polyethylene glycol (PEG), and diethyl ether. The results showed no significant differences in HBsAg values before and after Lipofundin addition, indicating that lipemia did not affect HBsAg detection by the CLIA method. However, lipid removal using centrifugation, PEG, and diethyl ether significantly reduced triglyceride and total cholesterol levels (p < 0.05) in lipemic samples that interfered with CLIA analysis. In conclusion, lipemia does not directly affect HBsAg screening results using the CLIA method; however, lipid removal procedures are effective and necessary for managing highly lipemic samples to maintain the accuracy and reliability of blood screening tests.
Hyperbaric Oxygen Therapy for Traumatic Brain Injury: A Review Of History, Development, Current Techniques, and Future Directions Wiwoho, Yudi Yuwono; Sadikin, Abdul Halim; Jusuf, Ahmad Aulia; Mulyawan, Wawan; Mudjihartini, Ninik; Ibrahim, Nurhadi; Jusman, Sri Widia A.; Sadikin, Mohamad
Proceedings Book of International Conference and Exhibition on The Indonesian Medical Education Research Institute Vol. 9 No. - (2025): Proceedings Book of International Conference and Exhibition on The Indonesian M
Publisher : Writing Center IMERI FMUI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.69951/proceedingsbookoficeonimeri.v9i-.321

Abstract

Hyperbaric oxygen therapy (HBOT) has gained increasing attention as a potential adjunctive treatment for traumatic brain injury (TBI) patients. This narrative review discusses the historical background, current preclinical and clinical studies, and explores its underlying mechanisms from biomolecular, histological, and clinical perspectives. HBOT promotes neural recovery by improving oxygenation, preserving mitochondrial integrity, enhancing neurotrophic support and synaptic connectivity, mitigating secondary injury pathways (including oxidative stress, inflammation, and apoptosis), and promoting angiogenesis and vascular stability. These mechanisms have demonstrated improvements of motor, cognitive, and memory functions both in preclinical and clinical studies, although outcomes and treatment protocols vary. However, challenges remain regarding optimal protocols, patient selection, and adverse effects. Further high-quality clinical trials are required to define the optimal HBOT regimen are required.
Acetazolamide-mediated carbonic anhydrase inhibition suppresses human peripheral blood mononuclear cell proliferation via G1/S cell cycle arrest Mustofa, Syazili; Sadikin, Mohamad; Sarmoko
Acta Biochimica Indonesiana Vol. 9 No. 1 (2026): 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.145

Abstract

Background: Carbonic anhydrase (CA) regulates intracellular CO₂/HCO₃⁻ homeostasis and supplies carboxyl donors for the sixth step of de novo purine biosynthesis. Disruption of this step is predicted to impair nucleotide pool accumulation and arrest cell cycle progression. Objective: This study investigated whether CA inhibition by acetazolamide suppresses T lymphocyte proliferation. Methods: Human peripheral blood mononuclear cells (PBMCs) were stimulated with phytohemagglutinin (PHA, 1% v/v) or interleukin-2 (IL-2, 10 ng/mL). Acetazolamide was applied at 6.25–50 μM. Cell viability, DNA synthesis, and cell cycle distribution were assessed using WST-1 assay, BrdU incorporation, and propidium iodide flow cytometry, respectively. Results: Acetazolamide reduced PBMC viability and DNA synthesis dose-dependently in both PHA- and IL-2-stimulated cultures (p < 0.05). IL-2-stimulated cells showed greater sensitivity, with significant inhibition at 12.5 μM versus 25 μM for PHA-stimulated cells. Flow cytometry revealed G1/S arrest in all treated groups: S phase decreased from 8.52% to 3.82% (PHA) and from 1.27% to 0% (IL-2) at 50 μM, with G2/M uniformly suppressed to ≤0.57%. Conclusion: Acetazolamide suppresses PBMC proliferation through G1/S arrest, consistent with CA inhibition depleting CO₂/HCO₃⁻-dependent carboxyl donors required for de novo purine synthesis.
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 Astriani , Ranti Dwi Astutiati Nurhasanah ASTUTIATI NURHASANAH Bethy S. Hernowo Caroline T. Sardjono CAROLINE TAN SARDJONO Cicia Firakania, Cicia DARYL ROBERT WILLIAMS 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 Hermansyah Hermansyah Indra G. Mansur Indratmo, Muhammad Faris Jeanne A. Pawitan Jeanne Adiwinata Pawitan Juniarti . Jusman, Sri Widia Azraki Lailan Safina Nasution Lusinanto, Arfat Mohamad Saekhu Muchlis Ramli Nani Dharmasetiawani Ni Ken Ritchie, Ni Ken 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 Sadikin, Abdul Halim Saleh, Mgs. M. Irsan Samsuridjal Djauzi Sari, Dewi H. Sarmoko Sarmoko Sarsanti, Pungguri Ayu Nega 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 Syazili Mustofa Tiwuk Susantiningsih Tomohiko Yamazaki Trismilah Trismilah TRISMILAH TRISMILAH Trismilah, Uly A. Nikmah, Uly A. Wahono Sumaryono Wawan Mulyawan Wawan Mulyawan Wiwoho, Yudi Yuwono Yefta Moenadjat Yudianita Kesuma, Yudianita Yuhernita . Zen Hafy