Doctorate Research

The microbiota in the gut influences health and well-being. The gut bacteria play a role in host pathogenesis, current opinion is that certain microbiota components can have beneficial effects on gastroenteritis resistance, blood lipids, antitumor properties, lactose tolerance, and gastrointestinal immunity. Modification of the human intestinal microbiota has currently become an important objective of dietetics. This goal can be attempted in two ways: either to include in the diet a significant proportion of beneficial bacteria, mainly Bifidobacterium and Lactobacillus species, with the expectation that they will be able to colonize the intestinal tract (probiotics); or to give non-digestible carbohydrates, like fructo-oligosaccharides, which have shown an ability to promote the growth of desirable bacteria (prebiotics).

Ingestion of probiotics stabilizes the gut microbiota and normalization of the intestinal microbiota by probiotics could lead to modulation of the host immune system. In addition, probiotics such as lactic acid bacteria are recognized by specific receptors on the surface of phagocytic cells. Meanwhile, composition and metabolic activity of the gut microbiota are directly depending on dietary constituents including prebiotics. Recent data indicate that prebiotics may modulate the gut microbiota and gut-associated lymphoid tissue (GALT) as well as the systemic immune system.

The purpose of this research was to evaluate the effect of local synbiotic (Lactobacillus plantarum Mut7 dan sweet potato fiber) for modulating the gut microbiota and immune responses. Ninety-six male Sprague dawley rats will be used for this experiment. In the first phase, 48 rats will be used and divided into 4 groups. Group A1 will receive AIN-93 diet, Group A2 AIN-93+sweet potato, Group A3 AIN-93+Mut7, and Group A4 AIN-93+sweet potato+Mut7. And then in the second phase, 48 rats will be used to study the effect of Salmonella typhimurium infection on the gut microbiota and immune responses. In this phase, the rats will be divided into 4 groups. Group B1 AIN-93 diet + S. thypimurium, Group B2 AIN-93+sweet potato + S. thypimurium, Group B3 AIN-93+Mut7 + S. thypimurium, and Group B4 AIN-93+sweet potato+Mut7+ S. thypimurium. The dosage of L. plantarum Mut7 is 109 CFU/ml, meanwhile the dosage of S. thypimurium is 1011 CFU/ml. In this experiment, we will used DGGE to differentiate 16S RNA species amplified by PCR from caecal samples of rats, together with the classical plate-count enumeration of the main bacterial groups. We will use ELISA method to evaluate the immunostimulatory effect of local synbiotic. Several immune parameter will be analyzed i.e. serum IgM, IgG, and IgA, sIgA in intestinal fluid, production of cytokines such as TNF- and IFN- in Peyer’s Patch, phagocytic activity of the macrophage peritoneum, and also population of T cells (CD4+ and CD8+) and B cell (CD19) in spleen and MLN. If the modulating effect of this local synbiotic can be confirmed, a functional food product can be developed. This study is expected to support policy to use local food products to improve health.

Tempe is made from fermented soybean by fungi to produce products that have a special flavor. Physiologically Tempe has a better biological value than the base ingredients itself. All this time, people consume tempe which is made from yellow soybean as the basic material, which is imported from U.S. Black soybean, as one of the varieties of soybeans, has similarities with the yellow one. Not many studies that examine tempe that is made from black soybean. Therefore, we need research on black soybean tempe from the aspect of technology and health. This research will study the prospects of antioxidants from black soybean tempe to improve immunity digestive tract. To prove that, there will be a research plan consists of two stages: (1) Study the Global Aspect on The Effect of Black Soybean Tempe on The Immune System, that consist about: (a) study the effect of black soybean tempe consumption and its antioxidant compounds to proliferation of mice’s intestinal mucosal lymphocytes cell in vivo and study the effect of black soybean tempe consumption and its antioxidant compounds to immunostimulatory activity of immunoglobulin (IgA) in mice induced by Salmonella typhimurium and (b) the effect of black soybean tempe and antoxidant extract consumption on human lymphocyte cell proliferation. (2) Cellular mechanism from effect of black soybean tempe on immune system within body.

Hypotheses of this study are that black soybean tempe has antioxidant activity and can increase the digestive tract immunity. The purpose of this study is to find out the potential of antioxidants in black soybean tempe to improve digestive tract immunity. The specific aims are; (1) to find out the effect of black soybean tempe consumption and its antioxidant coumpounds to intestinal mucosal lymphocyte proliferation in vivo, and (2) to find out the effect of black soybean tempe consumption and its antoxidant compounds to IgA in vivo, (3) to find out the effect of black soybean tempe and antioxidant extract consumption on human lymphocyte cell proliferation, and (4) to find out cellular mechanism from the effect black soybean tempe on immune system.

Pre-conceptional nutrition is critical period for determining the pregnancy outcome. Pre-pregnancy body mass index or maternal body size before pregnancy as one of strongest predictor of pregnancy outcome has been well documented. However, recently still scarce evidence of pre-conceptional micronutrient status regard to pregnancy outcomes, especially with randomized control trial design. Most of evidence which provided from previous studies was elucidated from prospective cohort studies. There is still an open challenge to explore the possible advantages of MMN supplementation during preconceptional period on birth weight and neonatus immune response. Therefore, in this year we are conducting a randomized double blind community-based trial to examine the effect of multiple micronutrients (MMN) supplementation during pre-conceptional period and continued during pregnancy on iron status and pregnancy outcomes especialy birth weight and neonatus immune response.

In the first step we intended to develop an integrated pre-marriage service system as a mechanism for subjects recruitment. This system is called LADUNI (Layanan Terpadu Pranikah). In future, this service system can be adopted by Local Government as a new program. The development of this system have been supported by Local Government by preparing media of health education to newly married women, and training of local health and marriage office personnel. The field preparation for implementation of LADUNI was done in 9 Sub Districts in Probolinggo. The try out of the implementation of LADUNI have been conducted at July 1th 2010.

The next step (Year-2), we would like to continue the activities to examine the effect of MMN supplementation on maternal iron status at last trimester of pregnancy, birth weight and infant’s immune response. Because of supplementation of micronutrient was developed as a part of the premarriage program that had been developed in the selected sub district of Probolinggo, therefore in this step, we also would like to evaluate whether the service system can work properly. To achieve this additional objective, we used a qualitative approach.

A randomized double blind community-based trial had been conducting at District of Probolinggo East Java, with the newly married women as subjects. In step two (Year 2) we focus to follow up subjects who had been enrolled previously (in year-1), who is pregnant, until delivering their babies. The subjects had been randomly assigned into two groups: Group I: received multi-micronutrient (MMN), UNICEF-derived MMN containing 15 micronutrients at least 3 months before pregnancy and continued during pregnancy; and Group II : received iron and folate (IFA) containing 60 mg iron and 250 μg folic acid only during pregnancy.

During preconception period, supplements will be consumed every two days (one day intermittent) maximum for 6 months, and when the subjects enter pregnancy period, supplement will be given in daily dose. Sample size of 418 ~ 420 newly married women had been recruited at enrollment, and predicted approximately 190 pregnant women to fulfill 150 deliveries or complete observation (75 subjects in each group), with calculation as described previously in year-1. The primary outcome variables are maternal iron status, birth weight and neonatal immune response, performed by cord blood IgG and ratio of CD4/CD8. Social economic background, nutritional status (BMI and MUAC), dietary intake, iron status (hemoglobin, ferritin, transferrin receptor/TfR), vitamin A status, zinc status, and blood pressure has been measured as base line data in year-1. Placental weight will be measure as secondary variable as well as the other variables such as maternal weight gain, morbidity, the presence of edema, gestational hypertension, preterm delivery, low birth weight and postpartum hemorrhage, will also be recorded.

Base line data will be analyzed by using independent-t test for continuous variables and χ2 test for categorical variables. To assess between-group differences by adjusting some confounder variables, analysis of covariance (ANCOVA) will be used. Content analysis will be applied for qualitative data.

The diabetes mellitus (DM) increases all over the world & in Indonesia it is the fourth among the highest (WHO, 2008). The prevalence of DM in urban areas nationwide found to be 5.7%, and glucose tolerance (IGT) found prevalence of 10.2% in 2007 (Riskesdas, 2008). Hyperglycemia increases free radical, reactive oxygen species (ROS), immune suppression and severely of dysfunction of pancreatic ß cells.

Normalize hyperglicemia & preserve β cell damage still fails with availability therapy DM right now (Djokomoeljanto, 2007). Kefir has been selected as one alternative treatment or complemetary lowering blood glucose. Clinical findings showed that administration of kefir in diabetic patients as much as 200 ml / day lowered their blood glucose (Yusi, et al, 2005). Study in vivo, kefir reduced to delta of blood glucose as much as 50.3 mg/dL (Judiono, 2007) and reduced the lipid peroxidation, increased antioxidants (SOD, Catalase, GPx) and the new novel that kefir regenerated or restored pancreatic β cell, and reduced proinflamation cytokine such as IL1, IL6 and increased IL10 (Judiono, 2010) However, it is very limited bimolecular elaborate its mechanism as antidiabetic for human setting in either Indonesia or International. This purposes to find novel the effect supplementation of kefir probiotic on glycemic status and functional pancreatic β cell, immune response of T2DM patients.

The research roadmap will be conducting in three phases. First, in vivo study aims to determine kefir’s ant diabetic biomolecular mechanism with a focus on the cell pancreas (2010). Second, to prove clinically aspects of kefir as antidiabetic on T2DM patients (2011). Third, to conduct to study the efficacy and bioactivity kefir probiotic encapsulation in the form of dry supplement products (2012).

The Randomized Pretest – Posttest Control Group Design will be chosen, and 108 T2DM divide into 3 groups. First, they receives kefir as much as 200 ml/day (treatment ) with fasting blood glucose ≥ 126 – 200 mg/dL, and second receives kefir as much as 200 ml/day (treatment) with fasting blood glucose ≥ 200-300 mg/dL, third group receives standard of DM medicine from their doctor (control). Assesment will be conducted in three phases such as; before intervention (1st phase), intensive intervention during 1 month (2nd phase) and end of intervention (3rd phase). The parameters will be collected consist of (1) semi quantitative food frequency & recall questioner to asses food & nutrients intake, (2) anthropometrics to measure BMI & weight gain, fat body composition, (3) blood examination to analyze such as; glycemic measure by Enzimatic Glucocard II, Insulin & C-peptide level to measure pancreatic β cell and immune response (IL1, IL6, IL10, TNFα) measured by Elisa, antioxidants level (SOD, GPx, Cat) and Lipid Peroxidation will be measured by Elisa Method. Data will be proccessed with SPSS for windows V. 16 and it will perform descriptive and statistical analysis.