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Background: Development a granuloma model resembling latent tuberculosis in vitro is needed with a fast and efficient time to be used as an effective therapy. This study aimed to form efficient granulomas, increase cellular immunity and humoral immunity, and evaluate growth on media using recombinant protein antibody Ag38kDa, Rifampicin, and a combination of both. Peripheral Blood Mononuclear Cell (PBMC) in vitro is derived from a healthy individual separated from monocytes and lymphocytes. Materials and methods: Monocytes are matured into macrophages and then combined macrophages and lymphocytes to the Roswell Park Memorial Institute (RPMI) medium. Flow cytometry analysis was used to count the number of cells, and cytokine levels were measured using ELISA. The result from the treatment was planted on the Lowenstein-Jensen medium. Results: Granulomas-like aggregates was formed after one-day post-infection with Mycobacterium tuberculosis (M.tb). A significant increase in immune response occurred in the number of macrophages, Th1, and Tregs in the combination group compared to the Mtb infection group. The number of Th2 and Th17 cells in the combination group was compared with the control but not significantly. TNF-α cytokine levels increased in the combination group compared to Mtb infection, while in IL-4, we found between all groups, there was no significant difference. Bacterial colonies on culture in the Lowenstein-Jensen medium were only seen in positive controls. Conclusion: Our study concluded that administration of a combination between Ag38kDa recombinant antibody and rifampicin could inhibit granuloma formation and enhance immune response.

Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is the etiology of an outbreak Covid-19. SARS-CoV-2 has a structural part consisting of spike glycoprotein, nucleoprotein N, membrane M and envelopes small membrane pentamer E. Immunoinformatic approach epitope analysis is developed to identify both weak and robust epitopes. Our study aims to identify several epitopes present in the spike glycoprotein, envelope, and membrane protein from the SARCoV-2 surface, with the help of insilico approach that highly potential as vaccine candidates. Analysis of antigeninicity was performed with the Kolaskar and Tongaonkar Antigenicity software. Epitope Mapping was analyzed using Linear Epitope Prediction Bepired. The structure of proteins with epitope regions was visualized by software Pyrex and PyMOL. Conserve analysis was performed using bio edit software. HLA mimicry was analyzed through HLAPred software. Molecular docking between the epitope with HLA I and HLA II was validated by Chimera and PyMOL software. The toxicity test for candidate vaccine peptides was carried out using ToxinPred software. Our study found seven potential epitope candidates as vaccine candidates. The seven epitopes were derived from spike proteins (5 epitopes), envelope proteins (1 epitope), and membrane proteins (1 epitope). All epitope codes are conserved and are not the same as HLA in Humans. The docking test results show a value with low affinity so that a strong bond can provide a high immune response. Toxicity tests show that all epitopes are non-toxic and safe to use as vaccine ingredients. Seven peptides from the spike, envelope, membrane protein that showed potential as vaccine candidates against Covid-19.

Bladder cancer is the ninth most common type of cancer occurring in men. Screening for bladder cancer is often difficult due to limited proper modality and symptoms that is mild and unspecific. Leaving it untreated leads to greater morbidity and mortality. Therefore, finding a simple biomarker for screening purpose is necessary. Cytokeratin (CK) has been studied extensively as cancer biomarker. Detectable soluble CK was found in body fluid, including urine, and histological specimen using immunoassays. The level of its expression is mainly affected by cell proliferation, apoptosis, and cellular stress. CK 14, which is type I CK, was detected in the basal layer of epithelial cells and specific for squamous type of epithelial cell. CK 14 analysis in bladder cancer has revealed numerous functions such as predicting the development of bladder cancer, differentiating cancer origins, stratifying bladder cancer patients based on disease severity, and predicting bladder cancer recurrence. Its potential to become excellent biomarker makes it subject of interest in research development.

Introduction: Previous studies have shown 37.8 kDa pili subunit protein of Vibrio cholerae and 49.8 kDa pili subunit protein of Shigella flexneri can act as an adhesion molecule to initiate infection. These molecules also have the ability to agglutinate blood. The present study assessed mucosal and systemic immunity following vaccination using 37.8 kDa V. cholerae and protection against S. flexneri. Subjects and Methods: Haemagglutination test was performed after purification of V. cholerae protein, followed by an anti-haemagglutination test. The intestinal weight and colony count were used to validate the protective effect on balb/c mice which were divided into the naive group, Shigella-positive control group, Vibrio-positive control group, V. cholerae infected-Vibrio-vaccinated group and S. flexneri-infected-Vibrio-vaccinated group. Th17, Treg, interleukin (IL) IL-17A, β-defensin and secretory-immunoglobulin A (s-IgA) were also measured to determine the systemic and mucosal immunity after vaccination. Results: The haemagglutination and anti-haemagglutination tests showed that the 37.8 kDa protein could inhibit 49.8 kDa of the S. flexneri pili subunit. Decreased intestinal weight and colony count of vaccinated group compared to naive group also support cross reaction findings. Vaccination also generates higher level of Th17, Treg, IL-17A, β-defensin and s-IgA significantly. Conclusions: 37.8 kDa subunit pili can act as a homologous vaccine candidate to prevent V. cholerae and S. flexneri infection.

Development a granuloma model resembling latent tuberculosisMonocytes are matured into macrophages and then combined macrophages and lymphocytes to the Roswell Park Memorial Institute (RPMI) medium. Flow cytometry analysis was used to count the number of cells, and cytokine levels were measured using ELISA. The result from the treatment was planted on the Lowenstein-Jensen medium.Granulomas-like aggregates was formed after one-day post-infection withOur study concluded that administration of a combination between Ag38kDa recombinant antibody and rifampicin could inhibit granuloma formation and enhance immune response.

Objective: To investigate the performance analysis of immunocytochemistry using 47 kDa polyclonal antibody compared with cytology in urine of patients hematuria symptom. Patients and methods: Patients with a suspicion of bladder tumor based on workup of hematuria were prospectively collected. The voided urine was performed for cytology and test with 47 kDa polyclonal antibody. Results: This study involved 39 patients consisting of 35 (89.74%) male and 4 (10.25%) female patients. For male patients, they showed 6 (17.14%) of positive cytology cases and 25 (71.42%) of positive immunocytochemistry cases. For female patients, they showed 0 (0.00%) of positive cytology cases and 2 (50.00%) of positive immunocytochemistry cases. The sensitivity and specificity value for immunocytochemistry using polyclonal antibody to the 47 kDa were 100.00% (95% CI = 54.07%–100.00%) and 36.36% (95% CI = 20.40%–54.88%), respectively. The positive and negative predictive value for immunocytochemistry of polyclonal antibody to the 47 kDa were 22.22% (95% CI = 8.62%–42.26%) and 100.00% (95% CI = 73.54%–100.00%), respectively. In conclusion, urine detection with 47 kDa polyclonal antibodies has higher sensitivity value and negative predictive value (100.00%) than urine cytology. Conclusion: This technique becomes an alternative early detection of bladder cancer for high-risk populations which can be easily and specifically carried out in primary care in Indonesia. Keywords: Bladder, Polyclonal antibody, Hematuria, Proteomic, Diagnostic performance

Although the AMV and AMS vaccine candidates have similar characteristics as hemagglutinin and adhesive molecules, there are differences in molecular weight.The research aims to determine the immunological cross-reaction between AMS and AMV.Antihemagglutination test used the anti-adhesion molecular antibody AMS. Next, we examined the immune response that has to be linked with protectivity. The model of the research uses MLIL. The sample separated the mice into four groups, and each group had five mice. The first group was the negative control group. The second group was given AMV and infected withAn antihemagglutination cross immune response, intestinal weight, the number of bacterial colonies, and other findings were found to be significant (The 49.8 kDa·MW protein subunit of the

Background: Covid-19 has become pandemic in the World, including Indonesia. Our last study showed that HSF could serve as an immunomodulator. Using the exact search, we found that the most immuno-dominant SARS-COV2 epitope, namely A spike protein epitope, B envelope protein epitope, and C membrane protein epitope, we concise to be HF Materials and Methods: We used to post only control design study and mice as an animal model. The research divided mice into four groups, and the first group as control received PBS as a placebo. The second, three, and last four groups gave HF, HSN, and HFHSN (combine HF and HSN). All of the regiment enters the mouth with a special sonde to reach the gastrointestinal organ. We gave HF every week three times and HSN once a day. After administration regiments for a long three weeks, we sacrificed the mice. We evaluated cellular immune responses that are Th-2, Th-17, and NK cells. We check for humoral immune response, TGF-β,IL-17A, IL-4, IgG,IL-4, β-defensin, and s-IgA. Results: Highest profile cellular immunity HF, HSN, and HFHSN were NK cell, Th-2 and Th-17, and the last NK cell, respectively. After that which in humoral immunity, the domination response IgG and IL-4 were HF. But HSN and HFHSN dominated for s-IgA and β-defensin production. By using the study Bio-Informatica, we found HF. Conclusion: If the results of this study are continued to the clinical trial level, it is necessary to recommend additional markers such as CTL (s-IgA and β-defensin in lung tissue)and CPE assay.

Vulvovaginal candidiasis occurs in 70 - 75% of women. About 40 - 45% of them experience vulvovaginal candidiasis for the second time. Approximately 70 - 89% of these infections are caused by Candida albicans. Candida albicans forming biofilm are reported to have a higher resistance to most anti-fungal drugs, including fluconazole. Objective: To determine the effects of propolis ethanolic extract (PEE) on biofilm formation and genes expression level of EFG1 on C. albicans biofilm. Methods: C. albicans biofilm formation was grown on SDB + 2% glucose and added with PEE with a concentration of 0% (control group) and 2.5, 5, 7.5, 10 and 12.5% (treatment group). Then, C. albicans biofilm formation was incubated at 37 °C for 48 hours. Biofilm inhibition was tested by the crystal violet assay. Analysis of genes expression level of EFG1 was performed using Real Time-PCR and calculated by absolute quantification method. Results: The results of the PEE effect on biofilm inhibition percentage showed a decrease in biofilm formation with the increase in PEE concentration given (p = 0.001), and gene expression level analysis showed that EFG1 (p = 0.000) expression level decreased with the increase in PEE concentration given. Conclusion: Propolis ethanolic extract could reduce biofilm formation and down-regulate the expression of EFG1 in C. albicans biofilm formation.

Previous studies have shown 37.8 kDa pili subunit protein of Vibrio cholerae and 49.8 kDa pili subunit protein of Shigella flexneri can act as an adhesion molecule to initiate infection. These molecules also have the ability to agglutinate blood. The present study assessed mucosal and systemic immunity following vaccination using 37.8 kDa V. cholerae and protection against S. flexneri.Haemagglutination test was performed after purification of V. cholerae protein, followed by an anti-haemagglutination test. The intestinal weight and colony count were used to validate the protective effect on balb/c mice which were divided into the naive group, Shigella-positive control group, Vibrio-positive control group, V. cholerae infected-Vibrio-vaccinated group and S. flexneri-infected-Vibrio-vaccinated group. Th17, Treg, interleukin (IL) IL-17A, β-defensin and secretory-immunoglobulin A (s-IgA) were also measured to determine the systemic and mucosal immunity after vaccination.The haemagglutination and anti-haemagglutination tests showed that the 37.8 kDa protein could inhibit 49.8 kDa of the S. flexneri pili subunit. Decreased intestinal weight and colony count of vaccinated group compared to naive group also support cross reaction findings. Vaccination also generates higher level of Th17, Treg, IL-17A, β-defensin and s-IgA significantly.37.8 kDa subunit pili can act as a homologous vaccine candidate to prevent V. cholerae and S. flexneri infection.

This study aims to determine the effect of statins and LPS antibodies administration combined with E. coli intraperitoneal injection toward proinflammatory mediators (TNF-?, hs-CRP, PCT, and MDA) urea, BUN, and creatinine) and liver function (SGPT, SGOT, and total bilirubin) in sepsis rats. This study used 30 male white rats (Rattus novergicus strain Wistar) divided into 6 groups induced by E. coli bacteria combined with statins, LPS antibody (Ab-LPS), and a combination of statins+Ab-LPS. Examination of proinflammatory mediators (TNF-?, hs-CRP, PCT, and MDA) was carried out by the ELISA method through the examination of biochemical markers namely renal function (urea, BUN, and creatinine) and liver function (SGPT, SGOT, and total bilirubin) were analyzed by using the autoanalyzer. The data obtained were statistically analyzed using the T-test. The result shows that E. coli+statins gave significant changes on the MDA level at 0th hour administration and TNF-? and hs-CRP at 3rd hour administration. The E. coli+Ab-LPS treatment showed significant changes at the 0th hour administration for MDA, creatinine, and total bilirubin and at the 3rd hour administration for MDA. The E. coli+statins+Ab-LPS treatment showed significant changes in MDA, creatinine, and total bilirubin at 0th hour administration and on creatinine and total bilirubin at 3rd hour administration.

BACKGROUND: The development of an oral typhoid fever vaccine need more effective and having high-efficacy in preventing typhoid fever. The use of liposomes as a vaccine vehicle can be formulated to target a specific location or trigger the release of antigens on its target. β-Glucan derived from Candida albicans cell wall as immunoadjuvant can strengthen the immune response and increases the protection against Salmonella Typhi bacterial invasion. AIM: This study aimed to determine the immune response in typhoid fever mice by administering a combination of AdhO36 S. Typhi liposome vaccine with β-Glucan and determine the protectivity to inhibit bacterial colonization in typhoid fever. METHODS: Mice were divided into five groups include negative and positive control also treatment group. IL-12 was evaluated after 4-h immunization while the other (was IL-12, IL-10, Th1 (IL-2), Th2 (IL-4), and the protective test against bacterial invasion) evaluated after 96-h. RESULTS: IL-12 level in the combination of β-Glucan and AdhO36 groups showed significantly lower than infected groups (p = 0.034), whereas IL-10 level significantly increase (p = 0.0009). The percentage of Th-1 (IL-2) cells significantly lower than infected groups (p = 0.000), this also happened on the percentage of Th-2 (IL-4) cells that significantly lower than infected groups (p = 0.018). The protective test toward bacterial invasion showed no bacterial colonization in all tissues intestine, liver, spleen, and mesenteric lymph node. CONCLUSION: The administration of a combination of liposome containing β-Glucan from C. albicans and AdhO36 S. Typhi has a potential effect on cellular and humoral immune response.

Turmeric ( Curcuma longa ) as a medicinal plant has widely consumed by pregnant women. The absence of dose standardization and the regulation of turmeric consumption in pregnancy raise concerns, especially if taken in early pregnancy. Angiogenesis is the process of new blood vessel formation from previously existing blood vessels, and plays an important role in embryogenesis and placentation during pregnancy. Angiogenesis is regulated by angiogenic molecules, such as VEGF and VE-cadherin. Turmeric has antiangiogenic effects in which the crude extract is pharmacologically more potent compared to the pure curcumin form. This study aimed to determine whether a turmeric decoctum affects the expression of VEGF and VE-cadherin in chick embryos. Turmeric was extracted by the decoction and freeze dried methods to obtain turmeric decoctum powder. This was tested on embryonated chicken eggs, which were divided into four groups; control group (2% DMSO) and treatment groups receiving various doses of the turmeric decoctum (200 ppm, 300 ppm and 400 ppm). The eggs used were less than 7 days after oviposition and incubated for 16 hours prior to injection of the treatment solution in ovo to the center of the yolk, followed by reincubation for up to 48 hours. Intraembryonic VEGF and VE-cadherin expression were assessed by whole-mount immunohistochemistry and quantified using image analysis techniques. This study showed administration of turmeric decoctum up to 400 ppm had no effect on the expression of VEGF and VE-cadherin in chick embryos.

The incidence of methicillin-resistant Staphylococcus aureus (MRSA) infection keeps increasing in every part of the world. Currently, the infection prevalence of MRSA has reached 70% in Asia. In Indonesia in 2006 the prevalence was 23.5%; the infection prevalence of MRSA in RS Atmajaya Jakarta reached 47%, in RSUP Dr. Moh. Husin Palembang reached 46%, and RSUD Abdul Moeloek Lampung in 2013 reached 38.4%. MRSA is multiresistant to antibiotics and is hard to kill compared to most other negative gram bacteria. The purpose of this research is to find the lethal power and exposure of ultrasonic waves to kill MRSA, monitoring its ef-fects via changes in shape, size, structure and Gram staining as indicators. The observations were done mac-roscopically by culturing the MRSA in a petri dish filled with Chromagar MRSA medium, while the morpho-logical observations of MRSA were done by SEM, changes in the structure using TEM, and changes in the color of MRSA cells using Gram staining. Ultrasonic wave exposure, at a lethal power = 8.432 watt, killed a significant percentage of MRSA over the control (p = 0.000). The death indicators of the MRSA due to expo-sure to ultrasonic waves of various power were: changes in shape of MRSA affected by ultrasonic power (p = 0.005), changes in size is not affected by ultrasonic power (p= 0.470), the stain of MRSA cell staining from purple to pink affected by ultrasonic power (p = 0.000), all compared with the control. MRSA died due to ne-crosis, with physical evidence of the MRSA death such as mechanical stress marked by swollen MRSA cell, shift cell wall, crack and tears, cavitation marked by pieces of MRSA cell in the field of view due to explosions inside the cell, change to an irregular cell shape, and changes in color from black to transparent.

Nigella sativa extract (NSE) is known to be an effective antibacterial and immuno-modulating agent. This study aimed to demonstrate that treatment of Salmonella eterica subsp. eterica serovars typhimurium- infected mice with NSE would cause an increase in T-regulatory (T-reg) and Th2 cells and a decrease in the number of spleen bacterial colonies. Mice were randomly divided into five groups [control (T0), mice infected with S. typhimurium only (T1), and mice infected with S. typhimur ium and then treated with NSE 0.52, 5.2, or 52 mg/kg body weight NSE (N1, N2, and N3, respectively)]. Levels of T-reg and Th2 cells were determined via flow cytometry and the number of spleen bacterial colonies was determined by observation. The results showed that treatment with NSE 0.52, 5.2, and 52 mg/kg significantly increased the number of T-reg and Th2 cells (p

Background: Bacterial vaginosis is commonly experienced by women of reproductive age. The Nugent score is the gold standard for diagnosing bacterial vaginosis. Prebiotic Glucomannan hydrolysates (GMH) as a therapy in the treatment of bacterial vaginosis and has an immunomodulatory effect on the immune system and provides cellular immunity. The purpose of this study was to examine the effects of GMH and BV Gel on Nugent scores, Treg cell presentation, and TGF ? levels in bacterial vaginosis of women of childbearing age.Subjects and Method: This was an experimental study. A sample of 24 women aged 20 to 45 years old with bacterial vaginosis (Nugent score ?7) was divided into 4 groups: oral antibiotic group metronidazole (500mg), combination of GMH (300mg) and metronidazole, 5 ml BV Gel tube, and combination of GMH and BV Gel scores. The dependent variables were GMH and BV Gel administrations. The independent variables Nugent score, Treg presentation, and TGF-? level. Nugent score, Treg cell presentation, and TGF ? level were measured on day-0, day-11, and day-22. The data were analyzed by one way Anova.Results: The results of the analysis after treatment on day 22 showed that the GMH and BV gel were able to reduce Nugent scores, increase Treg cell presentation and TGF ? levels in bacterial vaginosis of women of childbearing age.Conclusion: GMH as an alternative therapy for bacterial vaginosis compared with antibiotic treatment can improve normal vaginal flora and stimulate the immune system in vitro and in vivo significantly.Keywords: Glucomannan Hydrolysates, BV Gel, Nugent sel Treg score, TGF ?Correspondence:Ambar Dwi Retnoningrum. Masters Program of Midwifery, Universitas Brawijaya, Malang, East Java. Email: adreambar@gmail.com. Mobile: +6281335743696.Indonesian Journal of Medicine (2018), 3(1): 33-43https://doi.org/10.26911/theijmed.2018.03.01.05

Streptococcal pharyngitis sequel such as Rheumatic Fever (RF) or Rheumatic Heart Disease (RHD) is an autoimmune response mediated by T cells and IgG. Since it is an autoimmune process, the result of bacterial culture as the gold standard of diagnosis often shows negative results. IgG against the 33 kDa OMP is considered as an important mediator in the process of these autoimmune diseases, so its presence in blood serum can be used as a diagnostic tool. The purpose of this study is to prove that 33 kDa OMP is one of the immunogenic parts of the Streptococcus Group A β-hemolytic, so it is expected that IgG anti-33 kDa OMP can recognize and respond the bacteria and to support the probability of the Streptococcus Group A β-hemolytic infection. This study was a laboratory experimental study with a control group design. Animal used was RattusNovergicus immunized with whole cell bacteria or 33 kDa OMP mixed with Complete Freund’s Adjuvant or Incomplete Freund’s Adjuvant. Polyclonal IgG was obtained by drawing blood serum from the animals after immunization with Streptococcus Group A β-hemolytic for 4 weeks (A; n = 5) and 8 weeks (B; n = 5) or immunization with OMP 33 kDa for 4 weeks (C; n = 5) and 8 weeks (D; n = 5) and also negative control group (E; n = 5). Immunological tests were done using Dot Blot assay, ELISA, and immunocytochemical examination. The data obtained was then evaluated with statistical tests Kruskal-Wallis, Mann-Whitney and Repeated ANOVA (p < 0.05). The result showed that there was a difference in humoral immune response (IgG) between the groups albeit the difference was not significant (p > 0.05). Dot Blot and immunocytochemical tests indicated that IgG anti-33 kDa OMP were able to recognize and respond the Streptococcus Group A β-hemolytic antigen. This study concluded that 33 kDa OMP was the immunogenic part of the bacteria and that IgG anti-33 kDa OMP could recognize and respond the Streptococcus Group A β-hemolytic bacteria.

Shigella flexneri is the most common causal agent of shigellosis. Its pili are composed of pili protein subunits. Adhesion molecules can be found on the pili and outer membrane proteins (Omp). A hemagglutination reaction can be used for screening of adhesion molecules. Objectives: The purpose of this study was to determine the molecular weight of the pili protein subunits and outer membrane proteins of S. flexneri that act as hemagglutinin proteins, and to prove whether there is a cross-reaction between antibodies against hemagglutinin pili protein subunits and outer membrane proteins of S. flexneri. Methods: Pili protein subunits were isolated using pili bacteria cutters, and the outer membrane proteins were solubilized and obtained using sodium dodecyl sulfate 0.05% as detergent for Omp isolation. The hemagglutination reaction used mice erythrocytes. The cross reactions between subunit pili proteins were conducted by Western blot and Dot blot. Results:. Antibodies against hemagglutinin sub unit pili protein 18 kDa responded to pili protein subunits 18 kDa; 23 kDa; 34 kDa; and 53 kDa; and Omp 23 kDa and 27 kDa. Omp and subunit pili proteins S. flexneri consists of several identical epitopes that were responsible for the similarity of the response profile in the cross-reactions of antibodies.

Objective To explore the possible causes of infertility that occurs in Mojokerto, East Java, Indonesia. Methods The study was conducted by collecting biographical data in general and through clinical examination of all infertile couples. Other collected data included profiles of bacteria found in the vagina and an antibiotic sensitivity test of bacteria that can be isolated. To determine the possibility of antibodies against sperm a western blot examination of bacterial proteins in fluid secretions obtained from the vagina was carried out first. Results The results show that microorganisms were identified from the female reproductive tract of infertile couples dominated by Staphylococcus aureus (S. aureus) (27%) and Escherichia coli (27%), and sensitivity tests of antibiotics vary greatly. One of the microorganisms particularly S. aureus that found in large number was done western blotting using s-IgA cervix uteri as an antibody recognized Omp's with molecular weight, 52 kDa, and 49 kDa. Conclusion One of the bacteria species S. aureus that infects the vagina and causes nonspecific vaginitis can provoke an adaptive immune response by producing s-IgA. Difference MW of OMP S. aureus can stimulate immune response among women of infertile couples.

Background Recently, atherosclerosis-associated disease has been reported simultaneously increased. Whereas, to date, no atherosclerosis vaccine is available. Since the epitope mimicry between malondialdehyde low-density lipoprotein (MDA-LDL) and arginine specific epitope gingipain (Rgp) on the Porphyromonas gingivalis vesicles has been reported, it raises an opportunity to employ the potency of P. gingivalis as an atherosclerosis vaccine. Objective To evaluate the potency of P. gingivalis vesicles to prevent atherosclerosis, by assessing MDA-LDL level, visceral fat, body weight, and aortic wall thickness, in rats model. Methods Five groups of rats (n = 10 per group), three treatment groups, one positive and negative control group were assigned and adapted with high fat diet for 8 weeks. The treatment groups were injected with P. gingivalis vesicles with and without adjuvant with four booster doses. The level of MDA-LDL serum, visceral fat, body weight, and aortic wall thickness were measured in the end of the course. Results Our present study found that decreased in MDA-LDL levels (p = 0.037) and aortic wall thickness (p = 0.016) were observed in rats treated with vesicles and adjuvants, but not with vesicles or adjuvants only, compared to negative control. Moreover, MDA-LDL levels in rats immunized with vesicles and adjuvants were significantly lower than healthy rats. However, body weight (p = 0.329 and visceral fat (p = 0.789) were not significantly different in all treatment groups compared to control. Conclusions Immunization with P. gingivalis vesicles and adjuvants significantly reduces MDA-LDL level and aortic wall thickness in rats model.

Objective: The objective of this study was to determine whether Ramadan fasting (RF) alters the ability of serum, peripheral blood mononuclear cells (PBMC) and macrophages to kill Mycobacterium tuberculosis (M. tb). Methods: Thirty healthy male volunteers who fasted during Ramadan participated in the study. Blood sampling was conducted seven days before and on the seventh and 21 st day of Ramadan. The following parameters were measured by ELISA: Complement C3, inducible nitric oxide synthase (iNOS), superoxide dismutase (SOD) levels in serum and PBMC. The killing ability of immune components from these volunteers was determined by measuring colony-forming units (CFUs). Results: The CFU in PBMC infected with M. tb (PIM) and macrophages infected with M. tb (MIM) decreased on the seventh day of RF compared with that observed before RF. The CFUs in serum infected with M. tb (SIM) increased and CFUs in MIM decreased on the 21 st day of RF compared with before RF. The levels of iNOS in serum and the CFUs in MIM decreased and the CFUs in SIM and PIM increased on the 21 st day of RF compared with the seventh day of RF. The levels of iNOS in PBMC significantly increased (p

Helicobacter pylori (H.pylori) is a gram-negative microaerophilic bacterium that is generally associated with the main cause of approximately 80% peptic ulcers in stomach. It is also linked to the development of the stomach cancer. The aim of this study was to characterize the sub-unit pili proteins of Helicobacter pylori with a molecular weight of about 49,6 kDa as a haemagglutinin protein and adhesion molecules on surface of mice gastric epithelial cells. The bacterium was firstly cultured on the plate of TSA-B (Trypticase Soy Agar with 5% Sheep Blood) to prepare the protein of interes using bacterial cutter. The isolated proteins were subsequently analyzed for haemagglutination capability using mice sentisized erithrocytes. SDS- PAGE and western blotting analyses were used to confirm the protein patterns and their immunological reactivities respectively. The both SDS-PAGE and western blotting analyses showed that a dominant protein with molecular weight of about 49,6 kDa was detected consistently and found to be potentially aglutinate the erythrocytes. Furthermore, this protein was observed to be adherence on mice gastric epithelial cells. This study has indicated that the sub-unit pili proteins of H. pylori with a molecular weight of about 49,6 kDa was found to be dominant and immunogenic. Further study is required to confirm the in vivo properties of the protein associated with pathogenesis of H. pylori infection.

Acinetobacter baumannii (A. baumannii) is a pathogenic bacterium that can cause nosocomial infection in hospitalized patients with many kinds of manifestations. All bacterial infection is initiated by adhesion of bacterium to host cells through certain virulence factor which are called adhesin proteins. The purposes of this study were to isolate the pili subunit hemagglutinin (HA) protein of A. baumannii and to prove whether it functioned as adhesin protein and could activate the production of s-IgA. Isolation of pili subunit protein, was done using SDS-PAGE to get the molecular weight (MW) of pili subunit protein and then hemagglutinated. Adhesion test was conducted using pili subunit HA protein which had the highest titer of hemagglutination test, then coated on enterocyte cells of mice at the doses of 400 µg, 200 µg, 100 µg, 50 µg, 25 µg, 12.5 µg, 0 µg (control), respectively. ANOVA analysis results of dose response method from pili subunit HA protein of A. baumannii showed a significant difference of the index among treatment doses (p = 0.000). It can be seen that lower dose of pili subunit HA protein with MW of 38.8 kDa of A. baumannii could increase adhesion index. The 38.8 kDa pili subunit HA protein had significant influence on the adhesion index (r = - 0.729, p = 0.000, R square = 0.532, p = 0.000). Measurements of s-IgA concentration using ELISA method. There were differences in the number of s-IgA by ELISA test based on the treatment given to the value of p = 0.027 (p

1 Siti Nurul Mubarokah, 2 I Ketut Gede Muliartha, 3 Edi Widjajanto, 4 Sanarto Santoso, 5 Sumarno Reto Prawiro Laboratory of Biochemistry, Medical Faculty of Malang Islamic University Malang Laboratory of Clinical Anatomy and Pathology, Medical Faculty of Brawijaya University Malang Laboratory of Clinical Pathology, Medical Faculty Brawijaya University Malang Laboratory of Microbiology, Medical Faculty of Brawijaya University Malang

Bacterial culture is currently not recommended for routine evaluation of H. pylori infection in humans. For this reason, serological techniques play a critical role in diagnosing H. pylori infection in humans, particularly for initial pre-endoscopy or pre-treatment screening in dyspeptic patients. However, several current “in-office” tests appear to be less accurate or would need further validation before being recommended for use in a primary clinical care level. Preparation of specific and immunogenic antigens is an important step to improve the sensitivity and specificity of serologic assays, particularly for confirming the peptic ulcer status. The purpose of this study was to use a sub-unit protein of H. pylori with a molecular mass of about 49.6 kDa for the evaluation of H. pylori infection in mice. Fifty mice were orally infected with live H. pylori, and a similar number of mice were only orally given sterile phosphate-buffered saline (PBS). This process was repeated three times with a three-day interval between each administration, and blood samples were collected before and after each infection. The sera were tested using dot blot and ELISA. Seroconversion was detected two weeks after infection, and ELISA showed 98% for both its sensitivity and specificity. This study has indicated that the 49.6 kDa subunit pili proteins recognized homologous antibodies against the microorganism. Further studies are required to confirm the reaction of this protein against serum originated from H. pylori-infected human.

Purpose: To identify a Shigella flexneri hemagglutinin outer membrane protein (OMP) as an adhesion factor and examine its ability to cross-react with the OMPs of other Shigella species. Methods : OMP was isolated from the bacterium S. flexneri after shaving the pili using a pili bacterial cutter in a solution of 0.5 % n-octyl-β-D-glucopyranoside. Hemagglutination was evaluated using mice erythrocytes. The molecular weight (MW) of the OMPs was profiled by SDS-PAGE. Adhesion molecules were identified by calculating the index of adhesion. Thereafter, antigen cross-reaction with antibodies was examined by Western blotting, Results : The S. flexneri adhesion molecule OMP has a MW of 28 kDa. An antibody against the 28 kDa S. flexneri OMP adhesion molecule cross-reacted with different MW OMPs from other Shigella species. Conclusion : This study showed that the 28 kDa S. flexneri OMP is a hemagglutinin adhesion protein, and antibodies against it can cross-react with the OMPs of other Shigella species. Keywords : Shigella flexneri OMP, Outer membrane protein, Protein adhesion, Antibody, Cross-reaction

This study aims to investigate whether the protein isolated from bladder cancer in an Indonesian population can produce the polyclonal antibody for clinical markers of bladder cancer. The participants in this study are bladder cancer patients and healthy persons who were approved for midstream portion urine collection. The inclusion criteria included bladder cancer patients who obtained hematuria examination, urine cytology, and initial therapy with transurethral resection of the bladder; a healthy volunteer who was approved for midstream portion urine collection but without hematuria history, bladder stones, and signs or symptoms of tractus urinarius infection. The procedure consisted of tissue preparation, tissue processing, isolation of membrane cell protein, monitoring of protein in membrane cell, production of polyclonal antibodies, and dot blot technique. A protein with 122 kDa molecular weight is present in epithelial cells of bladder cancer and the normal bladder. A protein with 69 kDa molecular weight is only present in the epithelial cell bladder of normal individuals. In addition, a protein with molecular weight of 47 kDa is only present in epithelial cells of bladder cancer. The minimal ratio of polyclonal antibody with antigen is 1:6400 of the antibody and 1:40 of antigen. Subsequently, this concentration was applied to detect proteins with 47 kDa only in several cancer tissues. Positive results in bladder cancer, but negative results in prostate cancer, rectal cancer, and breast cancer were found. The polyclonal antibody produced from 47 kDa protein subunit is sensitive and specific to detect bladder cancer and become an alternative biomarker for diagnosis and surveillance of bladder cancer.