HYPOGLYCEMIC EFFECT OF HIGH-RESISTANT STARCH ANALOG RICE THROUGH GLP-1 AND INSULIN OR HIGH-RESISTANT STARCH ANALOG RICE ATTENUATES BLOOD GLUCOSE LEVEL THROUGH ENHANCEMENT OF GLP-1 AND INSULIN

Objective: This study was to investigate the effect of analog rice (AR) on glucagon-like peptide-1 (GLP-1) and insulin serum levels, glucose transporter-2 (GLUT-2) expression, and fasting blood glucose (FBG) level in diabetic rats. Methods: Fifty male Wistar rats divided into the control group (n=10) and the experimental group. High-fat diet and streptozotocin were administered in experimental groups, which then divided into four equal groups (n=10, each) (negative control group, rice group, AR1 and AR2 group, given standard pellet, rice pellet, AR1 and AR2 pellet, respectively, for 6 weeks). GLP-1 and insulin serum levels were measured by enzyme-linked immunosorbent assay. The expression of GLUT-2 and the number of pancreatic β-cells observed using an immunohistochemistry method. Results: FBG levels in the AR1 and AR2 groups decreased, while the rice group remained. GLP-1 serum levels of the negative control and rice groups were not significantly different from the control group, while the AR1 and AR2 groups higher than the control group (p≤0.05). All the treatment groups had insulin serum levels significantly lower than control group (p≤0.05), except the AR1 group. The expression of GLUT-2 and the number of pancreatic β-cells in the treatment groups were less than the control group, but between treatment groups were not significantly different. Conclusion: AR significantly effective in reducing FBG level in diabetic rats through stimulation of increased GLP-1 and insulin serum levels serum levels but AR did not affect on the expression of GLUT-2

Detection of Hepatitis B Virus DNA among Abdominal Typhus Patients with Hepatitis B Virus Co-Infection in Tuban District Based on Nested PCR Technique

Typhoid fever is often known by the name of abdominal typhus that may cause liver disruption so that arise disease complication which was known as typhoid hepatitis. Detection of abdominal typhus generally using routine blood test techniques that based on antigen and antibody reactions which is take a long time to obtain positive results so that these test cannot diagnose quickly and accurately. The detection of this disease in Tuban district is only using simple routine blood tests (eg Widal), urine and feces; it is difficult to diagnose the disease if there is complication. Abdominal typhus patients in Tuban district never got a specific examination technique by Nested Polymerase Chain Reaction (Nested PCR) to detect the occurrence of hepatitis B virus co-infection. The purpose of this study to detect hepatitis B virus DNA among abdominal typhus patients with hepatitis B virus co-infection by using Nested PCR techniques. This study is an experimental laboratory research. Blood samples from 9 positive HBsAg samples, which was obtained from 30 abdominal typhus patients in R. Kusma Hospital of Tuban District. The method used in this study was a nested PCR using primers P1 and P2 for the first PCR also primers HBV1 and HBV2 for the second PCR. Research carried out at the Biology Laboratory of Universitas  Ronggolawe (Unirow) Tuban and Institute of Tropical Disease, Universitas Airlangga (ITD Unair) in Surabaya, from March to May 2009. The results showed out of 9 positive HBsAg samples there were 4 (44.44%) HBV DNA detected using primer pair P1 and P2 then 1 (11.11%) were detected using primer pair HBS1 and HBS2. The use of two pairs of primers in nested PCR can detect more HBV DNA, because of the negative PCR results using a single primer pair can be detected using another primer pair. Conclusion of the study is usage of nested PCR technique using two different primer pairs can detect more HBV DNA in abdominal typhus patients with hepatitis B virus co-infection as much as 55.55%. Keywords: hepatitis B virus DNA, abdominal typhus patients, Nested PC

Detection of Hepatitis B Virus (HBV) DNA among Blood Donors with HBsAg Positive in Tuban District Based on Nested Polymerase Chain Reaction Technique (Nested PCR)

Hepatitis B virus (HBV) infection is a health problem in the world including Indonesia. It is proven by increasing incident from year to year in detected cases of patients infected with HBV either acute, chronic or on liver cirrhosis, and mild HBV infection is often accidentally detected during blood tests. The blood donors in Tuban district only got serological examination for the detection of Hepatitis surface antigen (HBsAg) but have not been specifically got examination by Nested Polymerase Chain Reaction (Nested PCR) for the detection of HBV DNA. Examination of high number of samples by nested PCR could provide better result (positive). The purpose of this study is for the detection of HBV DNA by nested PCR technique in HBsAg positive blood donors in the Indonesian Red Cross (PMI) of Tuban District. This study is an experimental laboratory research. Blood samples from 13 HBsAg positive samples were obtained from 150 blood donors at Indonesian Red Cross (PMI) of Tuban District. The method used in this study was a nested PCR using primer pair 7 and 8 for first PCR also HBS1 and HBS2 for second PCR. Research was established in the biology laboratory of Universitas Ronggolawe (Unirow) Tuban and Institute of Tropical Disease, Universitas Airlangga (ITD-Unair) Surabaya from March to May 2012. Results showed out of 13 HBsAg positive samples there were 3 (3/13 = 23.08%) HBV DNA detected using primers P7 and P8 then 10 (10/13 = 76.92%) were detected using primers HBS1 and HBS2. Usage two primer pairs in nested PCR can detect all (100%) HBV DNA because the negative PCR results with a single primer pair can be detected with another primer pair. Keywords: hepatitis B virus DNA, blood donors, Nested PC

Searching for the best agarose candidate from genus Gracilaria, Eucheuma, Gelidium and local brands

Objective: To explore the potential of local agar of genus Gracilaria, Eucheuma, Gelidium and local brands as an alternative for imported agarose for DNA electrophoresis, and to examine their ability related to separation and migration of DNA fragments in DNA electrophoresis. Methods: Their performance at various concentrations were compared via an experimental study with a specific brand of imported commercial agarose used in molecular biology research. The measured variables were separation and migration during electrophoresis of a DNA fragment. Results: The local agar genus Gracilaria gigas, Gelidium, brand "B" and brand "S" could separate DNA fragments at a concentration between 1% and 2%, with an optimum concentration of 2% w/v, as good as a specific brand of imported commercial agarose. Conclusions: Their performance were very close to that of commercial agarose and can still be improved by further agar purification as well as by pH and sulfur control

Searching for the best agarose candidate from genus Gracilaria, Eucheuma, Gelidium and local brands

Objective: To explore the potential of local agar of genus Gracilaria, Eucheuma, Gelidium and local brands as an alternative for imported agarose for DNA electrophoresis, and to examine their ability related to separation and migration of DNA fragments in DNA electrophoresis. Methods: Their performance at various concentrations were compared via an experimental study with a specific brand of imported commercial agarose used in molecular biology research. The measured variables were separation and migration during electrophoresis of a DNA fragment. Results: The local agar genus Gracilaria gigas, Gelidium, brand "B" and brand "S" could separate DNA fragments at a concentration between 1% and 2%, with an optimum concentration of 2% w/v, as good as a specific brand of imported commercial agarose. Conclusions: Their performance were very close to that of commercial agarose and can still be improved by further agar purification as well as by pH and sulfur control

Hepatitis C Virus Genotyping from anti-HCV Negative Sera in Blood Donors

HCV transmission is commonly derived from blood transfusions. Some different aspects, such as location, infection prevalence, and genotype distribution, may affect the occurrence of HCV in blood donors. The blood donors have already been screened regularly for their anti-HCV serology, yet the test for HCV RNA has not been done yet. In this study, we aim to investigate the manifestation of HCV in Tuban by detecting HCV RNA from sera negative for HCV antibodies in blood donors. The blood donors from Tuban Red Cross Indonesia were recruited for a questionnaire interview and testing for HCV antibodies and HCV nucleic acids. Anti-HCV was serologically detected using ELISA. Nested PCR was used to amplify HCV-RNA in the NS5B and 5’UTR regions. The genotype or subtype of HCV is determined by direct sequencing followed by phylogenetic analysis. A total of 100 blood samples were collected. The HCV RNA positive rate was 6% in sera-negative anti-HCV blood samples. Furthermore, the genotyping resulted in 4 samples being dominantly HCV subtype 1c (66,67%); the other 2 samples were subtype 2a and type 1 (each counted as 1 individual, 16.67%, respectively). The serological test for HCV antibodies has been shown to be less sensitive than the nucleic acid amplification test. The detection of genotype 1c as a major HCV genotype circulating in the Tuban area may help to anticipate HCV transmissions and facilitate better medical treatment with respect to HCV carriers

Detection of single-nucleotide polymorphism Gap junction protein Beta-2 genes in deaf schoolchildren of javanese population in Surabaya, Indonesia

Background: Genetic factors account for about 50%–75% responsible for hearing loss. The existence of single-nucleotide polymorphism (SNP) as genetic factors that affect hereditary hearing loss. The widely studied SNP was the gap junction protein beta-2 (GJB2) gene encoding the gap junction beta-2 protein (connexin26) that found in cochlea and required to convert sound waves into electrical nerve impulse. This study was aimed to detect the SNP GJB2 gene of hereditary hearing loss patients from Javanese population in Surabaya, Indonesia. Methods: The design of this study was a cross-sectional, analytic observational. The participant was taken randomly among the students from a deaf School in Surabaya. The questionnaire was completed by the parents of the deaf children. Blood sampling was taken from venous peripheral blood. DNA was extracted and amplified on GJB2 gene area by polymerase chain reaction (PCR). The positive results of PCR were processed further for sequencing. The sequencing results were analyzed to detect the GJB2 gene SNP with reference sequence/rs-80338939. Results: A total of 22 children participated in this study; all were profound sensorineural hearing loss (SNHL). The hereditary hearing loss was obtained with fewer in five children (22.73%), who had a history of hearing loss in their family. It was compared to 17 children (77.27%) who had no family history of hearing loss. It was found that the nucleotide variation in nucleotide number 8473 of GJB2 gene as much as 3 (13.64%) out of 22 children in hereditary hearing loss patients in Deaf School Type B Surabaya. Conclusions: This study did not found any SNP GJB2 gene (rs-80338939) of hereditary hearing loss patients from the Javanese population in Surabaya, Indonesia. There was the nucleotide substitution G to A in nucleotide number 8473 of GJB2 gene, which indicated the change of amino acid code genetic code table (valine) to amino acid code genetic code table (isoleucine). It may as the cause of SNHL

Genotip virus hepatitis C pada penderita yang menjalani hemodialis sebagai populasi beresiko tinggi tertular virus hepatitis C di Surabaya, Indonesia

Infeksi Virus Hepatitis C (VHC) masih menjadi masalah kesehatan di dunia termasuk di Indonesia. Dari tahun ke tahun, tdar banyak tl:rdell:ksi pcngidap penderita infeksi kronik VHC di seluruh dunia. Virus Hepatitis VHC dapat ditularkan secam parenteral. I\;nderita penyakit yang sedang menjaiani hemodialisis merupakan salah satu kelc.'n1pok penderita yang beresiko tinggi tertular infeksi VHC. Genotip tertentu VHC dilaporkan ikut bcrperan pada perjalanan infeksi virus hepatitis tersebut. Saat ini diketahui ada 6 genotipVHC didunia dan masing-masing genotip terdiri dari beberapa sUbtipe dengan penyebaran yang berbeda pada daerah geografi yang berbeda. Dikemukakan bahwa pada keadaan akut infeksi VHC sering tanpa gejala, sehingga sekitar 85% dari penderita yang terinfeksi VHC menjadi kronis. Prevalensi infeksi VHC cukup tinggi dan bervariasi pada penderita yang sedang menjalani hemodialisasi di berbagai negara dan infeksi VHC ini akan memperpendek umur penderita yang sedang menjalani hemodialisis. Dengan mempertimbangkan hal-hal tersebut diatas, maka dirasakan perlu dilakukan penelitian Genotip Virus Hepatitis C pad a penderita yaog sedang menjalani hemodialisis sebagai populasi beresilw tinggi tertular VHC di Surabaya, Indonesia

METS-IR vs. HOMA-AD on Obese adolescents

Purpose : Obesity is associated with chronic low-grade inflammation in which is the key in the pathogenesis Insulin Resistance (IR) and Metabolic Syndrome (MetS). Homeostasis model assessment of insulin resistance (HOMA-IR) has been validated as a surrogate measure of IR. The combination of HOMA and adiponectin, known as HOMA-AD was proposed to measure IR in adults. However, study on these indicators in obese adolescents is still limited. This study aims to analyse METS-IR and HOMA-AD to determine MetS and IR in obese adolescents. Methods : A cross-sectional study was conducted on obese adolescents who looked healthy from secondary schools in Surabaya and Sidoarjo, East Java, aged 12-18 years. Subjects were selected randomly and grouped into 2, namely MetS and non-MetS based on IDF 2000. Anthropometric examination and blood measurements, such as fasting blood glucose levels, lipid profiles, insulin, and adiponectin level were carried out according to standards. HOMA-IR, HOMA-AD, AND METS-IR were calculated using formula. Spearman’s Rho correlation were conducted between assessment tools (METS-IR and HOMA-AD) to identify the correlation with MetS component (lipid profile, FBG, and blood pressures). A receiving operation curve (ROC) performed to find area under curve (AUC) and cut-off points based on the biggest Youden index. Result : A total of 250 subjects were enrolled the study, and found 103 subjects had MetS. METS-IR correlates with all lipid profile and blood pressures (p < 0.05). While HOMA-AD correlated with TG (r = 0.356, p = 0.000), systolic-BP (r = 0.188, p = 0.003), and HDL-c levels (r=-0.249, p = 0.000). Cut-off point for METS-IR to determines MetS in obese adolescents was ≥ 46.53 (sensitivity of 64.24% and specificity of 75.76%), while HOMA-AD was ≥ 0.43 (sensitivity of 71.52% and specificity of 59.60%). Cut-off point for METS-IR index to determines IR was ≥ 52.01 (sensitivity of 83.44% and specificity of 44.44%). Cut-off point for HOMA-AD to determine IR was ≥ 0.37 (sensitivity of 74.17% and specificity of 84.85%). Conclusion : METS-IR is better surrogate to determine MetS with cut-off point of ≥ 46.53, while HOMA-AD is better to determine IR with cut-off point ≥ 0.37 in obese adolescents