19 research outputs found
Development of Vaccine Candidates Against Mycobacterium Tuberculosis in 2019-2023
No full textABSTRACT With extensive use of the Bacillus Calmette–Guérin (BCG) vaccine, the global prevalence of Mycobacterium tuberculosis (MTB) remains. A number of vaccines proposed to cure and prevent tuberculosis (TB) infection are undergoing various stages of clinical trials. Although vaccine production is progressing, more attention is needed. A number of TB vaccines are currently undergoing clinical trials, most of which rely on a combination of proteins and/or adjuvants or recombinant viral vectors specific for MTB antigens. We tried to cover the range of TB vaccines in this study by analyzing their composition, the immunological responses they elicit, and the stages of clinical trials. To find out the Development Of Prospective Vaccines Against Mycobacterium Tuberculosis. This research uses a literature review, between August 2023 and November 2023, the authors of this literature review checked PubMed, Science Direct, Google Scholar, and other databases containing research findings or scientific articles. Only studies that met the above search criteria were included in the systematic review. Many recently developed tuberculosis vaccines are reportedly in the final stages of clinical trials, where they have significantly strengthened the immune system and even produced protection against the host. Immunization produced by vaccines that have successfully passed the initial stages of clinical trials is safe and effective, and can even surpass BCG in terms of immunity. Based on the description above, it can be concluded that many recently developed tuberculosis vaccines are reported to be in the final stages of clinical trials, where they have significantly strengthened the immune system and even produced protection against the host. Immunization produced by vaccines that have successfully passed the initial stages of clinical trials is safe and effective, and can even surpass BCG in terms of immunity. With the development of new TB vaccines that strengthen the body's immunity and create effective delivery mechanisms, hopes for TB treatment and prevention are increasing. The development of vaccine effects can be facilitated, in part, through the use of effective delivery mechanisms, which have also been used in TB vaccines. Keywords: Tuberculosis, Vaccine, Bacteria, Mycobacterium Tuberculosis, BC
Distribution of blaCTX-M, blaSHV, blaTEM genes in Extended Spectrum β-Laktamase Producing Klebsiella pneumoniae from Clinical Isolates in Jakarta
No full textKlebsiella pneumoniae is one of the ESKAPE pathogens (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter spp.) due to their high level of antibiotic resistance. Ceftriaxone is one of the cephalosporin antibiotic that functions inhibit bacterial cell wall synthesis and used for treating K. pneumoniae infections. Resistance to ceftriaxone in K. pneumoniae has been widely reported, with one contributing factor being the production of β-lactamase enzymes encoded by the genes blaCTX-M, blaSHV, and blaTEM. This study characterized the presence of these genes  in 12 clinical isolates of         K. pneumoniae and analyzed their correlation with phenotypic resistance to ceftriaxone. All isolates characterized with antimicrobial susceptibility testing (AST) and disk diffusion methods to evaluate the phenotypic production of blaCTX-M, blaSHV, and blaTEM. Molecular analysis using the polymerase chain reaction (PCR) method showed the genes blaCTX-M and blaTEM were detected in 11 isolates (91.67%), and blaSHV was found in 9 isolates (75%). The distribution pattern of the blaCTX-M, blaSHV and blaTEM resistance genes was present in 8 isolates (66.67%), with MIC values > 64 µg/mL. The presence of blaCTX-M, blaSHV, and blaTEM genes together in K. pneumoniae isolates represents a potential risk for resistance to other β lactam antibiotics
Cloning of pe11 (LipX, Rv1169c) gene of Mycobacterium tuberculosis Beijing strain to pcDNA3.1 plasmid vector
No full textTuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis. It is a persistent global health problem with a high mortality rate. Currently, TB is controlled by administering the Bacillus Calmette-Guerin (BCG) vaccine, but the effectiveness of its protection varies among individuals in a population. The pe/ppe gene family comprises a typical group of genes that play a role in avoiding the host immune response and inducing persistent TB infection. Based on in silico analysis, the pe11 gene has estimated immunogenicity and potential as a TB seed vaccine candidate. The pe11 gene from an Indonesian isolate of an M. tuberculosis Beijing strain was amplified by polymerase chain reaction (PCR) and inserted into the mammalian expression vector pcDNA3.1. The recombinant vector pcDNA3.1-pe11 was used to transform Top10 competent Escherichia coli. Clones from the transformation were subjected to colony PCR to confirm the direction of the insert. Sequencing was performed to confirm the correctness of the insert sequence. In this study, the pe11 gene was successfully cloned into the pcDNA3.1 vector in the correct direction to assure PE11 expression. No mutations were found in the pe11 gene insert, compared with the M. tuberculosis H37Rv sequence as the standard. A pcDNA3.1 vector containing the pe11 gene derived from an M. tuberculosis Beijing strain was successfully constructed
Detection of human bocavirus (HBoV) in children with acute respiratory infection (ARI) during the covid-19 transition period
No full textAcute respiratory infections (ARIs) are the highest cause of death in children in the world. Based on the 2018 Riset Kesehatan Dasar Nasional, ARI cases in Indonesia showed a prevalence of 4.4%, with the highest cases occurring in children. One of the new viruses first identified in 2005 in human nasopharyngeal samples is the human bocavirus (HBoV). HBoV is a single-strand DNA virus belonging to the Parvoviridae family. This study aimed to assess the prevalence of HBoV in children presenting with ARI during the transitional period of the Covid-19 era. HBoV detection was conducted using multiplex PCR and reverse hybridization methods on nasopharyngeal and oropharyngeal swab samples collected from symptomatic children. This study reported a prevalence rate of 4.94% for HBoV in 2022 and 5.04% in 2023. Furthermore, the study identified favorable detection rates for HBoV in children with ARIs as 14.81% in 2022 and 8.45% in 2023. These rates ranked 2nd and 5th highest among other pathogens detected in ARIs. Additionally, there was an increase in positive HBoV samples from 4 samples in 2022 to 6 samples in 2023, which was attributed to the relaxation of nonpharmaceutical Covid-19 interventions by mid-2022. HBoV was identified at a significant rate among children with ARI in Jakarta during the transitional phase of the Covid-19 era (2022-2023). Given its potential to induce severe ARI, HBoV necessitates heightened attention as an etiological agent
Evaluation of Tuberculosis Vaccine Candidate, pcDNA3.1-rpfD using Mycobacterial Growth Inhibition Assay (MGIA)
No full textResuscitation-promoting factor D (RpfD) is a protein involved in the resuscitation of dormant bacteria. A new tuberculosis vaccine carrying the rpfD gene has been successfully constructed, pcDNA3.1-rpfD. It was demonstrated that this vaccine exhibits cellular and humoral immune responses. Therefore, within this study, the efficacy of this new vaccine candidate was evaluated using mycobacterial growth inhibition assay (MGIA). MGIA is a functional assay that measures the complex host immune response, peripheral blood mononuclear cell (PBMC) and splenocyte from BALB/c mice against mycobacteria. With BACTECTM MGITTM 960 automated system, the effect of vaccination on bacterial growth was reported as a time to positivity (TTP) in hours. The mean of TTP from the vaccinated group (both pcDNA3.1-rpfD and BCG) was higher than the negative control group. These results suggest that pcDNA3.1-rpfD may be effective in controlling tuberculosis growth and may provide a clue for the development of the tuberculosis vaccine. In addition, despite previous evidence that IFNγ was essential for tuberculosis immunity, IFNγ (interferon gamma) production was found not to be correlated with mycobacterial inhibition. Therefore, these findings offer an alternative method to evaluate vaccine candidates than the assessment using IFNγ only
The Effect of Probiotic Supplementation in Cholestasis Liver Disease: A Systematic Review of Animal Studies
No full textGut microbiome is essential in maintaining metabolism, gut barrier homeostasis, inflammation, and hematopoiesis. Several factors affect gut microbiome composition, including genetics, lifestyle, external factors, and disease. Cholestasis liver disease promotes gut dysbiosis via abnormal bile production or flow to the intestine and disrupts the gut microbiome. This condition leads to intestinal leakage, which enables bacterial and endotoxin translocation to the liver through the portal vein. Bacterial translocation promotes inflammatory responses, which worsen liver damage in cholestasis. Moreover, probiotic supplementation in other diseases has been shown to preserve gut microbiome composition. While such studies have documented probiotics' beneficial effects, no adequate clinical trials support probiotics' potency as a cholestasis treatment. Hence, this systematic review aims to provide an in-depth analysis of probiotic supplementation as a therapy for cholestasis liver disease in animal models. The search strategies were conducted based on PRISMA methodologies based on various academic literature. The selected studies have shown improvements in bile acid metabolism, microbiota-gut-liver axis, gut epithelium integrity, liver damage and inflammation response, and liver fibrosis progression, which need to be confirmed in human clinical trials
Responses of Humoral and Cellular Immune Mediators in BALB/c Mice to LipX (PE11) as Seed Tuberculosis Vaccine Candidates
No full textA member of the pe/ppe gene family, lipX (pe11), is capable of directing persistent Mycobacterium tuberculosis and avoiding host immune responses. Some studies have indicated that LipX (PE11) can detect humoral antibodies in tuberculosis patients. Hence, information on immune mediators’ responses to this protein is essential to understand its protective efficacy against M. tuberculosis infections. This study aimed to examine the response of immune mediators to pCDNA3.1-lipX expression in vivo. In the experiment, pCDNA3.1-lipX was injected into BALB/c strain male mice aged between 6 and 8 weeks, and they were compared to groups injected with pCDNA3.1 and without injection. The injection was carried out three times intramuscularly every two weeks. Blood was taken retro-orbitally and used for humoral response analysis by Western blotting against LipX-His protein. Simultaneously, the splenocytes were cultured and induced with LipX-His protein for cellular immunity analyses. Our study showed that the recombinant DNA of pCDNA3.1-lipX induced a humoral and cellular immune response, especially in IL-4, IL-12, and IFN-γ, which are the primary cellular responses to M. tuberculosis infections. However, additional studies, such as a challenge study, are needed to strengthen the argument that this plasmid construction is feasible as a tuberculosis seed vaccine candidate
Antiviral Activity of Cynometra ramiflora Linn Leaves Extract Against Replication of Dengue Virus Serotype 2 on Huh 7.5 Cell In Vitro
No full textMobile Genetic Elements Contributing to Carbapenem Resistance in Acinetobacter baumannii: Current Insights
No full textAcinetobacter baumannii has become a major cause of hospital-acquired infections with the rapid development of resistance to multiple antibiotics, including critical carbapenems. This resistance challenge limits treatment options and increases morbidity and mortality. The genetic plasticity of A. baumannii facilitates the mobilization of resistance genes via mobile genetic elements (MGE). Addressing this crisis requires a deeper understanding of the mechanisms by which MGE propagates carbapenem resistance. This paper provides a solution by systematically reviewing recent research on the role of MGE in disseminating resistance genes. Following PRISMA guidelines, a comprehensive literature review was conducted across various databases. The review revealed that resistance mechanisms primarily involve carbapenem-hydrolyzing enzymes and MGE, such as integrons, transposons, insertion sequences, and plasmids. Notably, genes like blaOXA-23 and blaNDM are frequently mobilized by these elements, facilitating horizontal gene transfer and persistence. Understanding the mechanisms of MGE-mediated gene transfer is crucial for developing strategies to control the spread of antibiotic resistance in A. baumannii
Infectivity and viability of dengue virus infected hepatocytes cocultured with peripheral blood mononuclear cells from a healthy subject
No full textBACKGROUND Dengue virus (DENV) can infect and replicate in monocytes, resulting in antibody-dependent enhancement. The liver is the main target of DENV, and the infection mechanisms of DENV include direct cytopathic effects (CPEs) of the virus, mitochondrial dysfunction, and effect of cellular and humoral immune factors in the liver. This study was aimed to explore the infectivity of DENV and viability of human hepatocytes using Huh 7it-1 cells cocultured with peripheral blood mononuclear cells (PBMCs).
METHODS Huh 7it-1 cells were infected with dengue virus serotype-2 (DENV-2) New Guinea C strain at multiplicity of infection of 0.5 and 1 FFU/cell, and cocultured in vitro with and without adherent PBMCs. The infectivity of DENV was assessed by immunoperoxidase staining. The viability of Huh 7it-1 cells was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT, a tetrazole) assay and trypan blue staining. Data were statistically analyzed by ShapiroWilk and analysis of variance for normality significances.
RESULTS The result showed that addition of PBMCs to DENV-2 infected Huh 7it-1 cells decreased the infectivity of DENV (1537%). DENV-2 infection decreased the viability of Huh 7it-1 cells (15.520.8%). Despite the decrease in infectivity of DENV, the addition of PBMCs increased the Huh 7it-1 cells viability (4.510.2%).
CONCLUSIONS Addition of PBMCs to Huh 7it-1 cells that are infected with DENV-2 decreased the infectivity of DENV and increased Huh 7it-1 cells viability
