Random amplified polymorphic DNA analysis of Mycobacterium tuberculosis isolates resistant to Isoniazid in Indonesia

Background: M. tuberculosisis the most important etiological factor of tuberculosis. One of the factors that make TB hard to eradicate is the emergence of M. tuberculosisdrug resistance. Drug resistance in M. tuberculosisis attributed primarily to the accumulation of mutations in the drug target gene. Objectives: to analyze profile of Random Amplified Polymorphic DNA (RAPD) in M. tuberculosisisolates resistant to Isoniazid and found RAPD marker. Methods: seven Isoniazid resistant isolate of M. tuberculosisfrom Ma kassar, Indonesia strain were analyzed by RAPD method using primers OPN 02, OPN 09, OPN 20, BG 65, N 9, that amplification fragment DNA than as molecular marker. Results: The results of the present study showed high degree of polymerphism in theM.tuberculosisstrains in the population, and found that specific DNA fragment at Isoniazid resistant isolates using primer N 9 is 1450 bp as a marker. Conclusion: This study gives information about RAPD marker of M. tuberculosis strain to Isoniazid resistant.

The Role of Cytokine Interleukin-2, Transcription Factor of FoxP3 in the Immunological Regulation of Allergic Rhinitis

Allergic rhinitis (AR) is the inflammation of nasal mucosa due to the type 1 hypersensitivity reactions mediated by immunoglobulin E (IgE) and triggered by certain allergens. The latest concept in allergic disease is the role of regulatory T cells (Treg). Interleukin-2 enhances the function and survival of Treg to perform its function as a controller of effector for forming a tolerant system by suppressing and regulating the homeostasis system. Treg has a transcription factor FoxP3 which plays a role in developing major function of Treg and progression to produce IL-10 and TGF-?. The atopic diseases are caused by a deficiency of Treg. The new perspective is low-dose IL-2 therapy towards autoimmune disease and allergic inflammation. Low-dose IL-2 therapy requires further clinical studies to optimize the dose, time, and the schedule of the IL-2 treatment. FoxP3 has the potential to assist in evaluating the active process of immunological process, which cannot be evaluated by Th1 and Th2 markers, and FoxP3 can be a successful immunotherapy marker.

Analysis Concentration of <i>Toxoplasma gondii</i>on Anti-Toxoplasma IgG-IgM Antibody Levels, and the Outcomes of Pregnancy in Mice Balb/c

Introduction: Toxoplasma gondii is an obligate intracellular protozoan parasite that can infect any warm blood vertebrae, and if first trimester pregnant woman infected, it may cause abortion. The objective is to prove the effect of the Toxoplasma gondii concentration in anti-toxoplasma IgG-IgM antibody levels, and the outcomes of Balb/c mice pregnancies. Materials and Methods: The study was conducted in Balb/c mice with inclusion criteria, and was conditioned pregnant. The pathogen strains of Toxoplasma gondii tachyzoite injected intraperitoneally. The blood samples were taken serially to be tested for anti-toxoplasma IgG-IgM antibody levels. After the mice were injected with tachyzoite, they are assessed every day to observe their body weight, vaginal bleeding, and labor. Anti-toxoplasma IgG-IgM antibody levels examined using qualitative mouse IgG-IgM antibody ELISA KIT. Results: Anti-toxoplasma IgM antibody levels increased significantly after 24 hours of injection tachyzoites in all dose groups, and remained high through day 21. Anti-toxoplasma antibody IgG levels increased significantly after 72 hours post injection and remained elevated until day 21. The incidence of abortion is 100% in mice which injected tachyzoite levels 1 × 103 and 1 × 104, and the incidence of abortion approximately 2 - 4 days post injection. 100% of mice that were injected with tachyzoites 1 × 101 and 1 × 102 have labor at term. Physical anomaly was found in baby mice from mice that were injected with tachyzoite 1 × 102. Conclusion: There is a significant correlation between the concentrations of Toxoplasma gondii tachyzoite with anti-toxoplasma IgG-IgM antibody levels, and there is a significant relationship between the concentrations of tachyzoite with abortion.

The Analysis of the Effective Systemic Lidocaine Dosage on the Expression of HMGB1 mRNA on Mice with Sterile Musculoskeletal Injury

A severe injury can trigger an inflammation response and excessive response can cause multiple organ failure. HMGB1 is an early inflammation mediator in sterile injury and a late inflammation mediator in infection. It is an important mediator in severe sepsis and always associated with the severity of organ failure. Previous studies showed that the administration of systemic lidocaine could inhibit the expression of HMGB1 on septic mice with musculoskeletal injury. Nine male adult Balb/c mice were grouped by simple random sampling method into three groups of intravenous lidocaine injection dosages: 2 mg/kg, 3 mg/kg, 4 mg/kg. Musculoskeletal injury was done by breaking the left femoral bone in a close manner. Peripheral blood sampling was done 4 hours after injury and 2 hours after lidocaine therap. To evaluate the expression level of HMGB1 mRNA, RT-PCR was used. The result of our study showed that intravenous lidoaine administration on the 3 groups could decrease the level of HMGB1. In conclusion, lidocaine hold an important role in clinical diseases by inhibiting HMGB1.

Analysis of Forkhead Box Protein-3 (Foxp3) in Allergic Rhinitis Patients

Background: A new concept in understanding allergic diseases is regulatory T cells (Tregs), which control the immune reaction caused by Th2 cytokine production. Forkhead Box Protein 3 (FoxP3) is a marker that has a critical role in the development and function of Tregs. Some studies found differences in FoxP3 level and in Tregs capacity to control immune reactions in allergic diseases. The aim of this study was to investigate FoxP3 level in Allergic rhinitis (AR) patients compared to atopic and healthy/normal persons in Jakarta. Methodology: This study used observation to analyze the level of FoxP3 in AR, atopic and healthy/normal persons, and used ELISA to measure the FoxP3 level. Results: The study had sixty participants divided into three groups: 21 in the Normal group, 16 in the Atopic group, and 23 in the AR group. The mean FoxP3 levels were 0.81 ± 0.35 in the normal group, 3.42 ± 0.15 in the atopic group, and 3.40 ± 0.13 in the AR group. Statistical analysis with Mann-Whitney tests indicated significant differences, with AR and atopic groups having higher FoxP3 levels than the normal group, (p = 0.001), and no statistically significant differences between the AR and atopic groups, (p = 0.92). Conclusion: Our study results suggested that FoxP3 was active in the control of inflammatory processes due to allergies, and decrease level of FoxP3 indicated severe AR, but suggested another mechanism caused differences in the clinical phenotypes of AR and atopic patients, despite them having equally high levels of FoxP3.

Prediction of hereditary nonpolyposis colorectal cancer using mRNA MSH2 quantitative and the correlation with nonmodifiable factor

BACKGROUND Hereditary non-polyposis colon cancer is a dominantly inherited syndrome of colorectal cancer(CRC),with heightened risk for younger population.Previous studies link its susceptibility to the DNA sequence polymorphism along with Amsterdam and Bethesda criteria.However,those fail in term of applicability.AIM To determine a clear cut-off of MSH2 gene expression for CRC heredity grouping factor.Further,the study also aims to examine the association of risk factors to the CRC heredity.METHODS The cross-sectional study observed 71 respondents from May 2018 to December 2019 in determining the CRC hereditary status through MSH2 mRNA expression using reverse transcription-polymerase chain reaction and the disease’s risk factors.Data were analyzed through Chi-Square,Fischer exact,t-test,Mann-Whitney,and multiple logistics.RESULTS There are significant differences of MSH2 within CRC group among tissue and blood;yet,negative for significance between groups.Through the blood gene expression fifth percentile,the hereditary CRC cut-off is 11059 fc,dividing the 40 CRC respondents to 32.5%with hereditary CRC.Significant risk factors include age,family history,and staging.Nonetheless,after multivariate control,age is just a confounder.Further,the study develops a probability equation with area under the curve 82.2%.CONCLUSION Numerous factors have significant relations to heredity of CRC patients.However,true important factors are staging and family history,while age and others are confounders.The study also established a definite cut-off point for heredity CRC based on mRNA MSH2 expression,11059 fc.These findings shall act as concrete foundations on further risk factors and/or genetical CRC future studies.

lnterleukin-6 and interleukin-10 plasma levels and mRNA expression in polytrauma patients

Purpose： Host response to polytrauma occasionally has unpredictable outcomes. Immune response is a major factor influencing patient＇s outcome. This study evaluated the interaction of two main cytokines in immune response after major trauma, specifically interleukin-6 （IL-6） and interleukin-10 （IL-10）. Plasma level of these cytokines is determined by mRNA expression of these cytokines genes which may decide the outcome of polytrauma patients. Methods： This prospective multicenter trial held at four trauma centers enrolled 54 polytrauma patients [Injury Severity Score （ISS） ≥ 16]. Plasma levels and mRNA expression of IL-6 and IL-10 were measured for 5 days after trauma. Clinical evaluation was conducted to observe whether patients endured multiple organ dysfunction syndrome （MODS） and death. MODS evaluation was performed using sequential organ failure assessment （SOFA）. Trauma load which in this study is represented with ISS, plasma level, expression of cytokine genes and patient＇s outcome were examined with correlation test and statistical analysis. Results： The elevated IL-6/IL-10 ratio indicated increased activity of systemic inflammation response, especially pro-inflammation response which bears higher probability of progressing to MODS and death. The decline of IL-6/IL-IO ratio with heavy trauma load （1SS 〉 30） showed that compensatory anti- inflammation response syndrome （CARS） state was more dominant than systemic inflammatory response syndrome （SIRS）, indicating that malfunction and failure of immune system eventually lead to MODS and deaths. The statistical significance in plasma level of cytokines was found in the outcome group which was defined as bearing a low trauma load but mortality. Conclusion： The pattern of cytokine levels in inflammation response has great impact on the outcome of polytrauma patients. Further study at the genetic level is needed to investigate inflammation process which may influence patient＇s outcome.