Budiman Bela - Profile on Academia.edu (original) (raw)
Papers by Budiman Bela
Journal of Physics: Conference Series, 2017
Cell penetrating peptides (CPP) are one of the most attractive DNA delivery systems currently in ... more Cell penetrating peptides (CPP) are one of the most attractive DNA delivery systems currently in development. In this research, in silico CPP development was performed based on a literature study to look for peptides that induce endosome escape, have the ability to bind DNA, and pass through cell membranes and/or nuclear membranes with a final goal of creating a new CPP to be used as a DNA delivery system. We report herein the successful isolation of three candidate CPP molecules, which have all been successfully expressed and purified by NiNTA. One of the determinants of CPP success as a DNA carrier is the ability of the CPP to bind and protect DNA from the effects of nucleases. The DNA binding test results show that all three CPPs can bind to DNA and protect it from the effects of serum nucleases. These three CPP candidates designed in silico and synthesized in the prokaryote system are eligible candidates for further testing of their ability to deliver DNA in vitro and in vivo.
Development of an assay system for genotyping Mycobacterium leprae resistant to dapsone, rifampicin, and ofloxacin
Indian journal of leprosy, 2019
Penyusunan ulang asam amino protein Early (E) 7 Human Papillomavirus tipe 16 untuk meningkatkan keamanan vaksin terapeutik
Jurnal Biotek Medisiana Indonesia, 2019
Sintesis dan pengklonaan fragmen-fragmen DNA APOBEC3G ke dalam vektor pBluescript KS (-) = Synthesis and cloning of fragments APOBEC3G DNA Into Bluescript SK (-) Vector
Lactobacillus plantarumIS-10506 supplementation increases faecal sIgA and immune response in children younger than two years
Beneficial Microbes, Apr 19, 2019
The immature intestinal immune system in young children develops as it comes into contact with di... more The immature intestinal immune system in young children develops as it comes into contact with dietary and microbial antigens in the gut. Intestinal microbiota plays a significant role in host defence mechanisms as shown by inflammatory responses towards potential pathogens. We investigated the probiotic function of Lactobacillus plantarum IS-10506 of ‘dadih’ origin in modulating immune response in young children. We aimed to assess its effect on their immune response by assessing transforming growth factor-β1 (TGF-β1) and tumour necrosis factor-α (TNF-α) responses and faecal secretory immunoglobulin A (sIgA) titre in a randomised, double-blinded placebo-controlled trial in 12-24-month-old children (n=38). We used four treatment groups for a 90-day supplementation period: placebo (n=11), probiotic (n=9), zinc (n=8) and probiotic and zinc (n=10). Faecal sIgA, plasma TGF-β1 and TNF-α titre were evaluated using the enzyme-linked immunosorbent assay standard technique. Statistical analysis divided the results (pre/post treatment) into high (>1) and low (<1) ratios. The results showed that faecal sIgA titre increased in all treatment groups compared with the control (placebo) and significantly increased in the probiotic group (P=0.05). In addition, the TGF-β1 ratio in the zinc group was significantly higher (P=0.05) than that in the placebo group. We observed a significant positive correlation between TGF-β1/TNF-α and faecal sIgA (r=0.27, P=0.04). Post hoc test results revealed that zinc supplementation has a significant effect on body-weight gain. Taken together, probiotic L. plantarum IS-10506 supplementation stimulates TGF-β1, which in turn increases the production of sIgA, in line with the significant correlation between TGF-β1/TNF-α and faecal sIgA.
Analysis of Recombinant VP22-OCT4 Fusion Protein Subcellular Location in HepG2 Cell Culture
Advanced Science Letters, Aug 1, 2018
There are many studies ongoing about the remarkable potential of pluripotent stem cell as the fut... more There are many studies ongoing about the remarkable potential of pluripotent stem cell as the future regenerative therapy. Embryonic stem cell is the only source that has been studied well for it. However, many constraints arise regarding this matter like ethical problems and risk of cell transplantation rejection. Recently, there are many research undergone in exploring new approach through reprogramming somatic cell to become induced pluripotent stem cell (iPSC). OCT4 (Octamer-binding transcription factor 4) is one of the proteins that is responsible for the self-renewal of embryonic stem cell. Therefore, this study aimed to investigate the ability of OCT4 transcription factor, with the help of VP22 (viral protein), to be localized into adult somatic cells (HepG2), which in turn could reprogram it into iPSC. In this study, HepG2 cells are isolated with VP22-OCT4 recombinant fusion protein for 1 hour and 6 hours, which then followed by isolating with mouse monoclonal IgG primary antibody against OCT4 (Ab1) and goat anti-mouse IgG secondary antibody, FITC conjugate (Ab2) respectively for the indirect immunofluorescence staining function. Confocal microscope is utilized to observe the presence of immunofluorescence. The result of the experiment showed a significant difference between the control and the experimental groups for both incubation period (p = 0.005 for 1 hour and p = 0.000 for 6 hours). Moreover, there is also a significance difference between the group of HepG2 cells with VP22-OCT4 incubated for 1 hour and 6 hours (p = 0.044). In conclusion, VP22-OCT4 recombinant protein can be translocated inside the HepG2 cells under 1 hour and 6 hours of incubation periods.
Medical Journal of Indonesia, Jun 30, 2020
BACKGROUND Apolipoprotein B mRNA editing enzyme catalytic polypeptide-like-3G (APOBEC3G) can abol... more BACKGROUND Apolipoprotein B mRNA editing enzyme catalytic polypeptide-like-3G (APOBEC3G) can abolish HIV infection by inducing lethal mutations in the HIV genome. The HIV protein virion infectivity factor (Vif) can interact with APOBEC3G protein and cause its degradation. Development of a method that can screen substances inhibiting the APOBEC3G-Vif interaction is necessary for identification of substances that potentially used in anti-HIV drug development. In order to increase expression of recombinant APOBEC3G protein that will be used in APOBEC3G-Vif interaction assay, we developed an optimized APOBEC3G gene for expression in Escherichia coli. METHODS The gene coding APOBEC3G was codon-optimized in accordance with prokaryotic codon using DNA 2.0 software to avoid bias codons that could inhibit its expression. The APOBEC3G gene was synthesized and sub-cloned into pQE80L plasmid vector. pQE80L containing APOBEC3G was screened by polymerase chain reaction, enzyme restriction, and sequencing to verify its DNA sequence. The recombinant APOBEC3G was expressed in E. coli under isopropyl-β-D-thiogalactoside (IPTG) induction and purified by using nickel-nitrilotriacetic acid (Ni-NTA) resin. RESULTS The synthetic gene coding APOBEC3G was successfully cloned into the pQE80L vector and could be expressed abundantly in E. coli BL21 in the presence of IPTG. CONCLUSIONS Recombinant APOBEC3G is robustly expressed in E. coli BL21, and the APOBEC3G protein could be purified by using Ni-NTA. The molecular weight of the recombinant APOBEC3G produced is smaller than the expected value. However, the protein is predicted to be able to interact with Vif because this interaction is determined by a specific domain located on the N-terminal of APOBEC3G.
Journal of Medical Microbiology, Aug 1, 2008
An overview of the phenotypic and genotypic characteristics of multidrug-resistant Mycobacterium ... more An overview of the phenotypic and genotypic characteristics of multidrug-resistant Mycobacterium tuberculosis isolates from four Asian countries
Journal of Ophthalmic Inflammation and Infection, Apr 25, 2023
Objectives This study aimed to investigate the spectrum of ocular characteristics and viral prese... more Objectives This study aimed to investigate the spectrum of ocular characteristics and viral presence in the conjunctival swab of patients with COVID-19. Methods In this cross-sectional study, fifty-three patients were recruited from two COVID-19 referral hospitals in Jakarta (Cipto Mangunkusumo Hospital and Persahabatan Hospital) from July 2020 to March 2021. The inclusion criteria were patients who were suspected of or confirmed cases of COVID-19 with or without ocular symptoms. Demographic data, history of COVID-19 exposure, underlying medical condition, systemic symptoms, ocular symptoms, supporting laboratory results, reverse-transcriptase polymerase chain reaction (RT-PCR) of naso-oropharyngeal (NOP) swab and conjunctival swab were collected. Results Fifty-three patients who were suspected, probable or confirmed cases of Covid-19 were included. Forty-six out of 53 patients (86.79%) tested positive for either Covid-19 antibody rapid test or naso-oropharyngeal (NOP) swab. Forty-two patients tested positive for NOP swab. Fourteen out of 42 patients (33.33%) experienced symptoms of ocular infection including red eye, epiphora, itchy eyes, and eye discharge. None of these patients were tested positive for conjunctival swab. Two out of 42 patients (4.76%), who were tested positive for conjunctival swab, did not experience any ocular symptoms. Conclusions Establishing the relationship between Covid-19 infection, ocular symptoms, and presence of SARS-CoV-2 virus on the ocular surface proves to be challenging. In Covid-19 patients, ocular symptoms did not warrant a positive conjunctival swab result. On the contrary, a patient without ocular symptoms can also have detectable presence of SARS-CoV-2 virus on the ocular surface.
KnE Life Sciences, Mar 25, 2019
Developing vaccines based on viral-like particles (VLPs) can be beneficial. VLPs represent viral ... more Developing vaccines based on viral-like particles (VLPs) can be beneficial. VLPs represent viral antigens that resemble viruses in nature. Unlike viruses that can replicate inside their host causing infection, VLPs have no capability to replicate and cause infection due to the lack of a viral genome. These properties make VLPs a preferable vaccine rather than inactivated viruses. In this research, VLPs were produced using a baculovirus infection system. Upon infection in SF9 insect cells, recombinant baculovirus produces new baculovirus proteins as well as the influenza proteins hemagglutinin (HA), neuraminidase (NA), and matrix (M1) protein, which can auto assemble into VLPs. VLPs can be separated from baculovirus using an ultracentrifuge. The first step of VLP production is selection of the baculovirus with recombinant genes encoding influenza proteins. The obstacle in selecting recombinant baculovirus is that infected SF9 cells do not produce specific cytopathogenic effects. To assist in the observation of baculovirus production in SF9 cells, we developed a protein reporter system for recombinant baculovirus production. Enhanced green fluorescent protein (eGFP) was used as a reporter protein. The gene encoding eGFP was cloned into the same transient plasmid that also encodes HA, NA, and matrix protein M1. Those proteins were expressed under four different open reading frames. This plasmid was co-transfected with a linearized baculovirus genome. Structural proteins of baculovirus and VLP forming recombinant proteins as well as the reporter protein were produced. Structural proteins of the baculovirus together with the genome of the baculoviruscontaining recombinant genes would co-assemble, producing recombinant baculovirus. The recombinant proteins then assembled, producing VLPs, and expression of the reporter protein eGFP would be visible in the cytoplasm by fluorescence microscopy. Using a TALI cytometer, the percentage of cells that produced GFP was determined to be 3% of the total population, indicating that eGFP can be used to observe the production of recombinant baculovirus in SF9 cells. The infectivity of baculovirus How to cite this article:
Konstruksi plasmid penyandi protein fusi vp22 gag hiv1 egfp untuk pengembangan vaksin sub unit gag hiv 1 endogen dan analisis lokalisasi protein vp22 egfp pada sel vero = Construction of plasmid encoding vp22 gag hiv1 egfp fusion protein for endogenous gag hiv 1 sub unit vaccine development and a...
Paediatrica Indonesiana
Background The World Health Organization (WHO) recommends viral load (VL) monitoring for HIV pati... more Background The World Health Organization (WHO) recommends viral load (VL) monitoring for HIV patients on antiretroviral therapy (ART). However, availability of VL monitoring in low-income countries remains limited. Objective To investigate factors associated with virological failure in HIV-infected children treated without routine VL monitoring. Methods This cohort study was done in children living with HIV (CLHIV) registered at Cipto Mangunkusumo General Hospital from 2004 to 2021. Viral load monitoring was not routinely done. Subjects with at least one VL result after 6 months on ART were included in the study. Virological failure was defined as a VL of >1,000 copies. Subjects’ data were obtained from medical records, laboratory reports, and dispensing pharmacies. Statistical analysis was done following survival analysis with hazard ratio. Results There were 384 children who had at least 1 VL result after ART was initiated. Median age at diagnosis was 30 months. Length of follo...
Buku Kurikulum Program Pendidikan Dokter Spesialis Mikrobiologi Klinik
Perhimpunan Dokter Spesialis Mikrobiologi Klinik Indonesia, 2018
Journal of Health Epidemiology and Communicable Diseases, 2019
Indonesia has not conduct regular screening test of CMV infection due to the lack of seropositive... more Indonesia has not conduct regular screening test of CMV infection due to the lack of seropositive prevelance data information. However, seronegative CMV results is not an indicator of safe blood for transfusion, so that another test that serves as confirmation test for CMV DNA is required. The aim of this study is to obtain prevalence data of CMV IgG antibody positive, the prevalence of CMV DNA positive and to determine the effect of CMV IgG titers against CMV DNA in blood donors in UTD PMI DKI Jakarta. Cross-sectional method was used to test 113 blood donor samples which have met inclusion criteria. Screening for CMV IgG antibody was held using indirect method chemiluminescence immunoassay (ChLIA) by Liason® XL 10050 Chemiluminescence Analyzer and CMV DNA analysis using qPCR method for the detection of CMV UL 54 with a tool Roche Light Cycler 480 II. Results indicate positive prevalence of IgG CMV in 111 samples (98.23%), and negative CMV IgG in 2 samples (1.77%). Prevalence of...
Journal of Physics: Conference Series, 2017
View the article online for updates and enhancements. Content from this work may be used under th... more View the article online for updates and enhancements. Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.
Nucleic acid amplification of HIV-1 integrase sequence subtypes CRF01_AE and B for development of HIV anti-integrase drug resistance genotyping assay
Journal of Physics: Conference Series, 2017
To anticipate the potential use of anti-integrase drugs in Indonesia for treatment of HIV-1 infec... more To anticipate the potential use of anti-integrase drugs in Indonesia for treatment of HIV-1 infection, the development of a drug resistance genotyping assay for anti-integrase is crucial in identifying the genetic drug resistance profile of Indonesian HIV-1 strains. This experiment aimed to amplify a target region in the integrase gene of Indonesian HIV-1 subtypes CRF01_AE and B that contain genetic mutations known to confer resistance to anti-integrase drug. Eleven archived plasma samples from individuals living with HIV-1 were obtained from the Virology and Cancer Pathobiology Research Center for Health Service (VCPRC FKUI-RSCM) laboratory. One of the plasma samples contained HIV-1 subtype B, and the remaining plasma samples contained subtype CRF01_AE. The target regions for all samples were amplified through RT-PCR, with an annealing temperature of 55 °C, using the primer pair AE_POL 4086F and AE_POL 5232R that were designed by VCPRC FKUI-RSCM. The results of this experiment show that 18.2% (2/11) of the samples were successfully amplified using the one-step RT-PCR. While the primer pair was effective in amplifying the target region in the integrase gene sequence for subtype B (100%; 1/1), it had a low efficacy (10%, 1/10) for subtype CRF01_AE. In conclusion, the primer pair can be used to amplify the target region in Indonesian HIV-1 strain subtypes CRF01_AE and B. However, optimization of the PCR condition and an increased number of samples would help to determine an accurate representation of the efficacy of the primer pair.
Molecular and Cellular Biomedical Sciences
Background: Reprogrammed cell therapy has not been applied for clinical purposes due to the malig... more Background: Reprogrammed cell therapy has not been applied for clinical purposes due to the malignancy issue. The aim of this study was to design the recombinant vector of the transcription factors and analyze the effectiveness of cell-penetrating peptide delivering system for human primary fibroblast transfection to avoid the malignancy issue.Materials and methods: The constructions of CCAT/enhancer binding protein alpha (CEBPA), hepatocyte nuclear factor 4 alpha (HNF4A), nuclear receptor subfamily 1 group I member 2 (NR1I2) were confirmed with DNA digestion and sequencing. Breast reduction (BRED) and palate (PAL) tissue were used as human primary fibroblast sources. The transcription factors were delivered into BRED and PAL with recombination of avian leukosis sarcoma virus (ALSV), human immunodeficiency virus (HIV) matrix, and regulator of expression of virion proteins (Rev) (ALMR), tagged with enhanced green fluorescence protein (eGFP). Post-transfection cells were then cultivat...
Jurnal Veteriner
Development of human immunodeficiency virus type 1 (HIV-1) vaccines and anti-retroviral treatment... more Development of human immunodeficiency virus type 1 (HIV-1) vaccines and anti-retroviral treatmentis currently hindered by the lack of models representing prominent symptoms of HIV-1 infections seen in humans. Simian-human immunodeficiency virus (SHIV) was constructed to resolve the limitations of SIVmac model and has been used in nonhuman primate model ofviral infections, particularly infections by the close relatives of HIV-1. Macaca fascicularis and M. nemestrina are being developed as model HIV/AIDS, by using chimeric virus SHIV produced by replacing the nucleotide structure of cyclophilin A binding region, vif gene and nef of HIV-1 with cyclophilin A binding region, vif gene and nef from SIV. The research aims to study the model of HIV/AIDS on nonhuman primates PBMC in vitro using SHIV. In particular, the study aims to obtain information about the capability of SHIV replication in PBMC of M. fascicularis and M. nemestrina. Results showed a cytopathic effect (CPE) in the form of ...
Suatu Antigen Rekombinan Yang Merupakan Bagian Immunodominan Protein Utama HIV-1 Envelope, GAG, dan Polimerase Untuk Dimanfaatkan Dalam Uji Diagnostik HIV-1
Pensubklonaan gen penyandi protein Tat HIV-1 galur oyi pada vektor plasmid pQE-80L = Subcloning of protein encoding genes Tat HIV-1 strain oyi into pQE-80L plasmid vector
Universitas Indonesia, 2019
Journal of Physics: Conference Series, 2017
Cell penetrating peptides (CPP) are one of the most attractive DNA delivery systems currently in ... more Cell penetrating peptides (CPP) are one of the most attractive DNA delivery systems currently in development. In this research, in silico CPP development was performed based on a literature study to look for peptides that induce endosome escape, have the ability to bind DNA, and pass through cell membranes and/or nuclear membranes with a final goal of creating a new CPP to be used as a DNA delivery system. We report herein the successful isolation of three candidate CPP molecules, which have all been successfully expressed and purified by NiNTA. One of the determinants of CPP success as a DNA carrier is the ability of the CPP to bind and protect DNA from the effects of nucleases. The DNA binding test results show that all three CPPs can bind to DNA and protect it from the effects of serum nucleases. These three CPP candidates designed in silico and synthesized in the prokaryote system are eligible candidates for further testing of their ability to deliver DNA in vitro and in vivo.
Development of an assay system for genotyping Mycobacterium leprae resistant to dapsone, rifampicin, and ofloxacin
Indian journal of leprosy, 2019
Penyusunan ulang asam amino protein Early (E) 7 Human Papillomavirus tipe 16 untuk meningkatkan keamanan vaksin terapeutik
Jurnal Biotek Medisiana Indonesia, 2019
Sintesis dan pengklonaan fragmen-fragmen DNA APOBEC3G ke dalam vektor pBluescript KS (-) = Synthesis and cloning of fragments APOBEC3G DNA Into Bluescript SK (-) Vector
Lactobacillus plantarumIS-10506 supplementation increases faecal sIgA and immune response in children younger than two years
Beneficial Microbes, Apr 19, 2019
The immature intestinal immune system in young children develops as it comes into contact with di... more The immature intestinal immune system in young children develops as it comes into contact with dietary and microbial antigens in the gut. Intestinal microbiota plays a significant role in host defence mechanisms as shown by inflammatory responses towards potential pathogens. We investigated the probiotic function of Lactobacillus plantarum IS-10506 of ‘dadih’ origin in modulating immune response in young children. We aimed to assess its effect on their immune response by assessing transforming growth factor-β1 (TGF-β1) and tumour necrosis factor-α (TNF-α) responses and faecal secretory immunoglobulin A (sIgA) titre in a randomised, double-blinded placebo-controlled trial in 12-24-month-old children (n=38). We used four treatment groups for a 90-day supplementation period: placebo (n=11), probiotic (n=9), zinc (n=8) and probiotic and zinc (n=10). Faecal sIgA, plasma TGF-β1 and TNF-α titre were evaluated using the enzyme-linked immunosorbent assay standard technique. Statistical analysis divided the results (pre/post treatment) into high (>1) and low (<1) ratios. The results showed that faecal sIgA titre increased in all treatment groups compared with the control (placebo) and significantly increased in the probiotic group (P=0.05). In addition, the TGF-β1 ratio in the zinc group was significantly higher (P=0.05) than that in the placebo group. We observed a significant positive correlation between TGF-β1/TNF-α and faecal sIgA (r=0.27, P=0.04). Post hoc test results revealed that zinc supplementation has a significant effect on body-weight gain. Taken together, probiotic L. plantarum IS-10506 supplementation stimulates TGF-β1, which in turn increases the production of sIgA, in line with the significant correlation between TGF-β1/TNF-α and faecal sIgA.
Analysis of Recombinant VP22-OCT4 Fusion Protein Subcellular Location in HepG2 Cell Culture
Advanced Science Letters, Aug 1, 2018
There are many studies ongoing about the remarkable potential of pluripotent stem cell as the fut... more There are many studies ongoing about the remarkable potential of pluripotent stem cell as the future regenerative therapy. Embryonic stem cell is the only source that has been studied well for it. However, many constraints arise regarding this matter like ethical problems and risk of cell transplantation rejection. Recently, there are many research undergone in exploring new approach through reprogramming somatic cell to become induced pluripotent stem cell (iPSC). OCT4 (Octamer-binding transcription factor 4) is one of the proteins that is responsible for the self-renewal of embryonic stem cell. Therefore, this study aimed to investigate the ability of OCT4 transcription factor, with the help of VP22 (viral protein), to be localized into adult somatic cells (HepG2), which in turn could reprogram it into iPSC. In this study, HepG2 cells are isolated with VP22-OCT4 recombinant fusion protein for 1 hour and 6 hours, which then followed by isolating with mouse monoclonal IgG primary antibody against OCT4 (Ab1) and goat anti-mouse IgG secondary antibody, FITC conjugate (Ab2) respectively for the indirect immunofluorescence staining function. Confocal microscope is utilized to observe the presence of immunofluorescence. The result of the experiment showed a significant difference between the control and the experimental groups for both incubation period (p = 0.005 for 1 hour and p = 0.000 for 6 hours). Moreover, there is also a significance difference between the group of HepG2 cells with VP22-OCT4 incubated for 1 hour and 6 hours (p = 0.044). In conclusion, VP22-OCT4 recombinant protein can be translocated inside the HepG2 cells under 1 hour and 6 hours of incubation periods.
Medical Journal of Indonesia, Jun 30, 2020
BACKGROUND Apolipoprotein B mRNA editing enzyme catalytic polypeptide-like-3G (APOBEC3G) can abol... more BACKGROUND Apolipoprotein B mRNA editing enzyme catalytic polypeptide-like-3G (APOBEC3G) can abolish HIV infection by inducing lethal mutations in the HIV genome. The HIV protein virion infectivity factor (Vif) can interact with APOBEC3G protein and cause its degradation. Development of a method that can screen substances inhibiting the APOBEC3G-Vif interaction is necessary for identification of substances that potentially used in anti-HIV drug development. In order to increase expression of recombinant APOBEC3G protein that will be used in APOBEC3G-Vif interaction assay, we developed an optimized APOBEC3G gene for expression in Escherichia coli. METHODS The gene coding APOBEC3G was codon-optimized in accordance with prokaryotic codon using DNA 2.0 software to avoid bias codons that could inhibit its expression. The APOBEC3G gene was synthesized and sub-cloned into pQE80L plasmid vector. pQE80L containing APOBEC3G was screened by polymerase chain reaction, enzyme restriction, and sequencing to verify its DNA sequence. The recombinant APOBEC3G was expressed in E. coli under isopropyl-β-D-thiogalactoside (IPTG) induction and purified by using nickel-nitrilotriacetic acid (Ni-NTA) resin. RESULTS The synthetic gene coding APOBEC3G was successfully cloned into the pQE80L vector and could be expressed abundantly in E. coli BL21 in the presence of IPTG. CONCLUSIONS Recombinant APOBEC3G is robustly expressed in E. coli BL21, and the APOBEC3G protein could be purified by using Ni-NTA. The molecular weight of the recombinant APOBEC3G produced is smaller than the expected value. However, the protein is predicted to be able to interact with Vif because this interaction is determined by a specific domain located on the N-terminal of APOBEC3G.
Journal of Medical Microbiology, Aug 1, 2008
An overview of the phenotypic and genotypic characteristics of multidrug-resistant Mycobacterium ... more An overview of the phenotypic and genotypic characteristics of multidrug-resistant Mycobacterium tuberculosis isolates from four Asian countries
Journal of Ophthalmic Inflammation and Infection, Apr 25, 2023
Objectives This study aimed to investigate the spectrum of ocular characteristics and viral prese... more Objectives This study aimed to investigate the spectrum of ocular characteristics and viral presence in the conjunctival swab of patients with COVID-19. Methods In this cross-sectional study, fifty-three patients were recruited from two COVID-19 referral hospitals in Jakarta (Cipto Mangunkusumo Hospital and Persahabatan Hospital) from July 2020 to March 2021. The inclusion criteria were patients who were suspected of or confirmed cases of COVID-19 with or without ocular symptoms. Demographic data, history of COVID-19 exposure, underlying medical condition, systemic symptoms, ocular symptoms, supporting laboratory results, reverse-transcriptase polymerase chain reaction (RT-PCR) of naso-oropharyngeal (NOP) swab and conjunctival swab were collected. Results Fifty-three patients who were suspected, probable or confirmed cases of Covid-19 were included. Forty-six out of 53 patients (86.79%) tested positive for either Covid-19 antibody rapid test or naso-oropharyngeal (NOP) swab. Forty-two patients tested positive for NOP swab. Fourteen out of 42 patients (33.33%) experienced symptoms of ocular infection including red eye, epiphora, itchy eyes, and eye discharge. None of these patients were tested positive for conjunctival swab. Two out of 42 patients (4.76%), who were tested positive for conjunctival swab, did not experience any ocular symptoms. Conclusions Establishing the relationship between Covid-19 infection, ocular symptoms, and presence of SARS-CoV-2 virus on the ocular surface proves to be challenging. In Covid-19 patients, ocular symptoms did not warrant a positive conjunctival swab result. On the contrary, a patient without ocular symptoms can also have detectable presence of SARS-CoV-2 virus on the ocular surface.
KnE Life Sciences, Mar 25, 2019
Developing vaccines based on viral-like particles (VLPs) can be beneficial. VLPs represent viral ... more Developing vaccines based on viral-like particles (VLPs) can be beneficial. VLPs represent viral antigens that resemble viruses in nature. Unlike viruses that can replicate inside their host causing infection, VLPs have no capability to replicate and cause infection due to the lack of a viral genome. These properties make VLPs a preferable vaccine rather than inactivated viruses. In this research, VLPs were produced using a baculovirus infection system. Upon infection in SF9 insect cells, recombinant baculovirus produces new baculovirus proteins as well as the influenza proteins hemagglutinin (HA), neuraminidase (NA), and matrix (M1) protein, which can auto assemble into VLPs. VLPs can be separated from baculovirus using an ultracentrifuge. The first step of VLP production is selection of the baculovirus with recombinant genes encoding influenza proteins. The obstacle in selecting recombinant baculovirus is that infected SF9 cells do not produce specific cytopathogenic effects. To assist in the observation of baculovirus production in SF9 cells, we developed a protein reporter system for recombinant baculovirus production. Enhanced green fluorescent protein (eGFP) was used as a reporter protein. The gene encoding eGFP was cloned into the same transient plasmid that also encodes HA, NA, and matrix protein M1. Those proteins were expressed under four different open reading frames. This plasmid was co-transfected with a linearized baculovirus genome. Structural proteins of baculovirus and VLP forming recombinant proteins as well as the reporter protein were produced. Structural proteins of the baculovirus together with the genome of the baculoviruscontaining recombinant genes would co-assemble, producing recombinant baculovirus. The recombinant proteins then assembled, producing VLPs, and expression of the reporter protein eGFP would be visible in the cytoplasm by fluorescence microscopy. Using a TALI cytometer, the percentage of cells that produced GFP was determined to be 3% of the total population, indicating that eGFP can be used to observe the production of recombinant baculovirus in SF9 cells. The infectivity of baculovirus How to cite this article:
Konstruksi plasmid penyandi protein fusi vp22 gag hiv1 egfp untuk pengembangan vaksin sub unit gag hiv 1 endogen dan analisis lokalisasi protein vp22 egfp pada sel vero = Construction of plasmid encoding vp22 gag hiv1 egfp fusion protein for endogenous gag hiv 1 sub unit vaccine development and a...
Paediatrica Indonesiana
Background The World Health Organization (WHO) recommends viral load (VL) monitoring for HIV pati... more Background The World Health Organization (WHO) recommends viral load (VL) monitoring for HIV patients on antiretroviral therapy (ART). However, availability of VL monitoring in low-income countries remains limited. Objective To investigate factors associated with virological failure in HIV-infected children treated without routine VL monitoring. Methods This cohort study was done in children living with HIV (CLHIV) registered at Cipto Mangunkusumo General Hospital from 2004 to 2021. Viral load monitoring was not routinely done. Subjects with at least one VL result after 6 months on ART were included in the study. Virological failure was defined as a VL of >1,000 copies. Subjects’ data were obtained from medical records, laboratory reports, and dispensing pharmacies. Statistical analysis was done following survival analysis with hazard ratio. Results There were 384 children who had at least 1 VL result after ART was initiated. Median age at diagnosis was 30 months. Length of follo...
Buku Kurikulum Program Pendidikan Dokter Spesialis Mikrobiologi Klinik
Perhimpunan Dokter Spesialis Mikrobiologi Klinik Indonesia, 2018
Journal of Health Epidemiology and Communicable Diseases, 2019
Indonesia has not conduct regular screening test of CMV infection due to the lack of seropositive... more Indonesia has not conduct regular screening test of CMV infection due to the lack of seropositive prevelance data information. However, seronegative CMV results is not an indicator of safe blood for transfusion, so that another test that serves as confirmation test for CMV DNA is required. The aim of this study is to obtain prevalence data of CMV IgG antibody positive, the prevalence of CMV DNA positive and to determine the effect of CMV IgG titers against CMV DNA in blood donors in UTD PMI DKI Jakarta. Cross-sectional method was used to test 113 blood donor samples which have met inclusion criteria. Screening for CMV IgG antibody was held using indirect method chemiluminescence immunoassay (ChLIA) by Liason® XL 10050 Chemiluminescence Analyzer and CMV DNA analysis using qPCR method for the detection of CMV UL 54 with a tool Roche Light Cycler 480 II. Results indicate positive prevalence of IgG CMV in 111 samples (98.23%), and negative CMV IgG in 2 samples (1.77%). Prevalence of...
Journal of Physics: Conference Series, 2017
View the article online for updates and enhancements. Content from this work may be used under th... more View the article online for updates and enhancements. Content from this work may be used under the terms of the Creative Commons Attribution 3.0 licence. Any further distribution of this work must maintain attribution to the author(s) and the title of the work, journal citation and DOI.
Nucleic acid amplification of HIV-1 integrase sequence subtypes CRF01_AE and B for development of HIV anti-integrase drug resistance genotyping assay
Journal of Physics: Conference Series, 2017
To anticipate the potential use of anti-integrase drugs in Indonesia for treatment of HIV-1 infec... more To anticipate the potential use of anti-integrase drugs in Indonesia for treatment of HIV-1 infection, the development of a drug resistance genotyping assay for anti-integrase is crucial in identifying the genetic drug resistance profile of Indonesian HIV-1 strains. This experiment aimed to amplify a target region in the integrase gene of Indonesian HIV-1 subtypes CRF01_AE and B that contain genetic mutations known to confer resistance to anti-integrase drug. Eleven archived plasma samples from individuals living with HIV-1 were obtained from the Virology and Cancer Pathobiology Research Center for Health Service (VCPRC FKUI-RSCM) laboratory. One of the plasma samples contained HIV-1 subtype B, and the remaining plasma samples contained subtype CRF01_AE. The target regions for all samples were amplified through RT-PCR, with an annealing temperature of 55 °C, using the primer pair AE_POL 4086F and AE_POL 5232R that were designed by VCPRC FKUI-RSCM. The results of this experiment show that 18.2% (2/11) of the samples were successfully amplified using the one-step RT-PCR. While the primer pair was effective in amplifying the target region in the integrase gene sequence for subtype B (100%; 1/1), it had a low efficacy (10%, 1/10) for subtype CRF01_AE. In conclusion, the primer pair can be used to amplify the target region in Indonesian HIV-1 strain subtypes CRF01_AE and B. However, optimization of the PCR condition and an increased number of samples would help to determine an accurate representation of the efficacy of the primer pair.
Molecular and Cellular Biomedical Sciences
Background: Reprogrammed cell therapy has not been applied for clinical purposes due to the malig... more Background: Reprogrammed cell therapy has not been applied for clinical purposes due to the malignancy issue. The aim of this study was to design the recombinant vector of the transcription factors and analyze the effectiveness of cell-penetrating peptide delivering system for human primary fibroblast transfection to avoid the malignancy issue.Materials and methods: The constructions of CCAT/enhancer binding protein alpha (CEBPA), hepatocyte nuclear factor 4 alpha (HNF4A), nuclear receptor subfamily 1 group I member 2 (NR1I2) were confirmed with DNA digestion and sequencing. Breast reduction (BRED) and palate (PAL) tissue were used as human primary fibroblast sources. The transcription factors were delivered into BRED and PAL with recombination of avian leukosis sarcoma virus (ALSV), human immunodeficiency virus (HIV) matrix, and regulator of expression of virion proteins (Rev) (ALMR), tagged with enhanced green fluorescence protein (eGFP). Post-transfection cells were then cultivat...
Jurnal Veteriner
Development of human immunodeficiency virus type 1 (HIV-1) vaccines and anti-retroviral treatment... more Development of human immunodeficiency virus type 1 (HIV-1) vaccines and anti-retroviral treatmentis currently hindered by the lack of models representing prominent symptoms of HIV-1 infections seen in humans. Simian-human immunodeficiency virus (SHIV) was constructed to resolve the limitations of SIVmac model and has been used in nonhuman primate model ofviral infections, particularly infections by the close relatives of HIV-1. Macaca fascicularis and M. nemestrina are being developed as model HIV/AIDS, by using chimeric virus SHIV produced by replacing the nucleotide structure of cyclophilin A binding region, vif gene and nef of HIV-1 with cyclophilin A binding region, vif gene and nef from SIV. The research aims to study the model of HIV/AIDS on nonhuman primates PBMC in vitro using SHIV. In particular, the study aims to obtain information about the capability of SHIV replication in PBMC of M. fascicularis and M. nemestrina. Results showed a cytopathic effect (CPE) in the form of ...
Suatu Antigen Rekombinan Yang Merupakan Bagian Immunodominan Protein Utama HIV-1 Envelope, GAG, dan Polimerase Untuk Dimanfaatkan Dalam Uji Diagnostik HIV-1
Pensubklonaan gen penyandi protein Tat HIV-1 galur oyi pada vektor plasmid pQE-80L = Subcloning of protein encoding genes Tat HIV-1 strain oyi into pQE-80L plasmid vector
Universitas Indonesia, 2019