Enzymatic synthesis of phenolic acid glucosyl esters to test activities on cholangiocarcinoma cells.

While glycoside hydrolase family 1 (GH1) enzymes mostly catalyze hydrolysis reactions, rice Os9BGlu31 preferentially catalyzes transglycosylation to transfer a glucosyl moiety to another aglycone moiety to form a new glycosylated compound through a retaining mechanism. In this study, Os9BGlu31 was used to synthesize eight phenolic acid glucosyl esters, which were evaluated for activities in cholangiocarcinoma cells. The transglycosylation products of Os9BGlu31 wild type and its mutant variants were detected, produced on a milligram scale, and purified, and their structures were characterized by NMR spectroscopy. The transglycosylation products were evaluated by antioxidant and anti-proliferative assays, followed by an anti-migration assay for the selected phenolic acid glucosyl ester. Os9BGlu31 mutants produced higher yield and activity than wild-type enzymes on phenolic acids to produce phenolic acid glucosyl esters. Among these, gallic acid glucosyl ester (ß-glucogallin) had the highest antioxidant activity and anti-proliferative activity in cholangiocarcinoma cells. It also inhibited the migration of cholangiocarcinoma cells. Our study demonstrated that rice Os9BGlu31 transglucosidase is a promising enzyme for glycosylation of bioactive compounds in one-step reactions and provides evidence that ß-glucogallin inhibits cell proliferation and migration of cholangiocarcinoma cells. KEY POINTS: • Os9BGlu31 transglucosidases produced phenolic acid glucosyl esters for bioactivity testing. • Phenolic acid glucosyl esters were tested for cytotoxicity in cholangiocarcinoma cells. • ß-Glucogallin displayed the highest inhibition of cholangiocarcinoma cell growth.

Microencapsulation of Ruellia tuberosa L. Aqueous Root Extracts Using Chitosan-Sodium Tripolyphosphate and Their In Vitro Biological Activities.

The current study aims to perform microencapsulation of R. tuberosa L. extracts using chitosan crosslinked to sodium tripolyphosphate (NaTPP) as wall materials by spray drying and to analyze their in vitro biological activities. The influence of manufacturing conditions, like pH, chitosan concentration, and stirrer time, was assessed. Results showed that microcapsules prepared in pH 4 with a concentration of 0.1% (w/v) chitosan, and 90 min stirring time had 51.80% encapsulation efficiency and high in vitro biological activity. These were shown by high in vitro alpha amylase inhibition and antioxidant activity with IC50 values of 50.65 µg/mL and 123.97 µg/mL, respectively. Releases of the bioactive compounds in microcapsules of R. tuberosa L. were carried out on phosphate buffer medium pH 2.2 and pH 7.4 with times release of 30, 60, 90, and 120 min. The bioactive compounds were released in pH 2.2 in 120 min at 2.48%. At pH 7.4, the active ingredients were more easily released, by 79.90% in 120 min. The microcapsules' morphology showed a rough surface with spherical forms and the average sizes were 53.41 µm. This study supports the essential role of microencapsulation in improving plant extracts with reserved biological activities.

Transient receptor potential vanilloid (TRPV) channels: Basal properties and physiological potential.

Transient receptor potential vanilloid (TRPV) channels are TRP homologs and have been classified into six subfamilies. They are unique mediators of sensory signals with multiple physiological effects and are potential targets for developing new therapies targeting human diseases. TRPV channels play crucial roles in normal physiological processes, and their dysfunction has been implicated in various disease states. Several small-molecule compounds, such as TRPV1 and TRPV3 antagonists, have been developed as novel analgesic agents. A better understanding of the physiological functions of TRPV channels would lead to progress in life science. In this review, we focus on various functions of TRPV channels, including pain sensing, temperature sensing, and metabolic control, as well as summarize the basal properties and pathophysiological contributions of six TRPV channels. Moreover, we discuss the pharmacological effects of endogenous and exogenous ligands on TRPV channels and related diseases.

The Expression of Myoz1 and ApoB is Positively Correlated with Meat Quality of Broiler Chicken.

Broiler chicken (Gallus gallus) is a source of animal protein with a high nutritional content. The purpose of this study was to evaluate the quality of broiler chicken meat (Gallus gallus) by analyzing its nutritional value, genetic profile, and protein level. The chicken meat samples were obtained from four different districts in Malang city, Indonesia. We analysed the proximate composition of chicken meat to detect its nutrition content. Furthermore, we have examined the sequence of the Myoz1 gene as well as the level of ApoB proteins in the same meat. The nutritional analysis of chicken meat showed that in the four locations different levels of protein, ash, water, and lipids were observed. The Myoz1 gene of femur chicken broilers from the second and third districts has five and twenty-one substitution mutations, respectively. The ApoB expression level in locations 2 and 3 was higher than that in the other districts. In conclusion, Myoz1 and ApoB levels were correlated with the nutritional content and quality of broiler chicken meat.

Histopathological Profiles of Rats (Rattus norvegicus) Induced with Streptozotocin and Treated with Aqueous Root Extracts of Ruellia tuberosa L.

Diabetes mellitus (DM) is a serious worldwide health threat since the number of people with DM is forecasted to grow annually. Thus, effective and affordable treatment is urgently needed. Our previous studies used n-hexane and hydroethanolic root extracts of Ruellia tuberosa L. which significantly affected diabetic rats. In this study, aqueous R. tuberosa L. root extracts were used as treatments for the diabetic rat model and their effects were evaluated. Diabetes was generated by the administration of streptozotocin (STZ) at 20 mg/kg within 5 sequential days. Male Wistar rats were orally treated with the extracts and standard drug (metformin 200 mg/kg) and vehicle every day for 4 weeks. Hypoglycemic effects were assessed for normal, diabetic control, standard, and extract-treated groups. Histopathology was also carried out for the pancreatic, hepatic, and kidney tissues. The progression of diabetes was considerably diminished after extract treatment. In treated rats, the highest dose of extract induced a decline in blood glucose and serum malondialdehyde (MDA) levels at 25% and 35%, respectively. Furthermore, aqueous extract of R. tuberosa L. treatment decreased MDA levels in the pancreas by 12%. Histologic examination of the organ tissues of diabetic rats showed deteriorating alterations. After treatment, histopathological damages to the tissues and cells were reversed. The results of the experiments recommend that aqueous extract of R. tuberosa L. has antidiabetic effects on STZ-induced diabetic rats; nevertheless, a higher dose of the aqueous extracts is needed to achieve more significant results.

Omics-Based Analytical Approaches for Assessing Chicken Species and Breeds in Food Authentication.

Chicken is known to be the most common meat type involved in food mislabeling and adulteration. Establishing a method to authenticate chicken content precisely and identifying chicken breeds as declared in processed food is crucial for protecting consumers' rights. Categorizing the authentication method into their respective omics disciplines, such as genomics, transcriptomics, proteomics, lipidomics, metabolomics, and glycomics, and the implementation of bioinformatics or chemometrics in data analysis can assist the researcher in improving the currently available techniques. Designing a vast range of instruments and analytical methods at the molecular level is vital for overcoming the technical drawback in discriminating chicken from other species and even within its breed. This review aims to provide insight and highlight previous and current approaches suitable for countering different circumstances in chicken authentication.

Purple Sweet Potatoes from East Java of Indonesia Revealed the Macronutrient, Anthocyanin Compound and Antidepressant Activity Candidate.

INTRODUCTION: The development of new antidepressant is crucial to overcome the remission rate limitation. Anthocyanin on purple sweet potatoes (PSP) from East Java cultivar previously demonstrated a behavioural effect. However, the certain mechanism and the nutritional compound need further exploration. AIM: This study aimed to characterize macronutrient content, amino acids, anthocyanin, and revealed the potential of PSP from East Java-Indonesia as antidepressant agent through D2-dopamine receptor (D2DR). METHODS: This study was characterized the macronutrient content using proximate analysis. The amino acids were analysed using Ultra-Performance Liquid Chromatography (UPLC) and High-Performance Liquid Chromatography (HPLC). Anthocyanin was identified using Ultra High-Performance Liquid Chromatography (UHPLC). Molecular docking was conducted to predict the interaction between anthocyanins and D2 dopamine receptor. RESULTS: We were found the predominance of water on proximate analysis. Alanine was demonstrated as the highest content of amino acid. Cyanidin, cyanidin-3-O-glucoside and peonidin-3-O-glucoside were identified as major anthocyanin content. Molecular docking was showed that cyanidin bound to similar binding site with dopamine on D2DR with stronger interaction than cyanidin-3-glucoside. CONCLUSION: Current study was indicated that cyanidin as major anthocyanin from purple sweet potatoes has potential health beneficial as antidepressant potential candidate.

The dual-specificity protein kinase Clk3 is essential for Xenopus neural development.

During vertebrate development, the formation of the central nervous system (CNS) is initiated by neural induction and patterning of the embryonic ectoderm. We previously reported that Cdc2-like kinase 2 (Clk2) promotes neural development in Xenopus embryos by regulating morphogen signaling. However, the functions of other Clk family members and their roles in early embryonic development remain unknown. Here, we show that in addition to Clk2, Clk1 and Clk3 play a role in the formation of neural tissue in Xenopus. clk1 and clk3 are co-expressed in the developing neural tissue during early Xenopus embryogenesis. We found that overexpression of clk1 and clk3 increases the expression of neural marker genes in ectodermal explants. Furthermore, knockdown experiments showed that clk3 is required for the formation of neural tissues. These results suggest that Xenopus Clk3 plays an essential role in promoting neural development during early embryogenesis.

The amelioration of T2DM rat femoral bone achieved by anti-osteoporosis of caprine CSN1S2 protein through bone morphogenetic protein signaling pathway.

We investigated the potential anti-glycation and anti-osteoporosis properties of Caprine milk CSN1S2 protein on the serum AGEs and sRAGE level, osteogenic factors expressions, femoral bone mesostructure, histomorphometry, and hydroxyapatite crystals changes in T2DM rats. Varying doses of Caprine milk CSN1S2 protein (0, 375, 750, and 1500 mg/kg BW) were used to treat the control and T2DM rats. We measured AGEs and sRAGE level; RUNX2, OSX, BMP2, and Caspase-3 expressions in rats using ELISA and immunohistochemistry, respectively. The mesostructure and histomorphometry of femoral bone was analyzed using SEM Microscope and BoneJ software, then hydroxyapatite crystal size was determined using SEM-XRD. T2DM rats showed a high level of AGEs and a low level of sRAGE, the RUNX2, OSX, and BMP2 expression was down regulated, BV, BV.TV, Tb.Th, Tb.Sp, increased and SMI levels declined, respectively. Vice versa, after administration of the CSN1S2 protein to T2DM rats, improvement in all levels of molecular and cellular markers was achieved. In the CSN1S2 highest dose, AGEs level declined and sRAGE level elevated in T2DM rats. The 375 and 750 mg/kgBW of CSN1S2 protein was able to upregulate the RUNX2, OSX, and BMP2 expression in T2DM rats, thus improving the normalization of osteoclasts and osteoblasts number. The whole dose of CSN1S2 triggered the thickening of trabecular bone wall, granule formation, and normalized the trabecular thickness (Tb.Th) parameter of T2DM rats. The hydroxyapatite crystal size was increased in the highest dose of CSN1S2-treated T2DM rats. This study indicated that CSN1S2 protein had a protective effect against osteoporosis in the T2DM rat bones by means of glycation pathway inhibition, bone histomorphometry and mesostructure improvement via bone morphometric protein signaling.

Black rice cultivar from Java Island of Indonesia revealed genomic, proteomic, and anthocyanin nutritional value.

Black rice is considered to be functional food containing anthocyanins as bioactive compounds. This study examined the genomic and proteomic patterns in local black rice from Java Island, Indonesia, with attention to the mechanism of anthocyanin synthesis. Three kinds of black rice from Java Island, including black rice from East Java (BREJ), black rice from Central Java (BRCJ), and black rice from West Java (BRWJ), were studied in comparison to white rice (WREJ) and red rice (RREJ). Genomic profiling was done by simple sequence repeat (SSR) analysis, and sequencing of red coleoptile (Rc) and glycosyltransferase (GT) genes, followed by in silico analysis. Total anthocyanin was investigated by ultra-high performance liquid chromatography- diode array detector (UHPLC-DAD). The proteomic profiles were determined by liquid-chromatography and mass spectrometry of tryptic peptides. The SSR profiles showed a specific band in each black rice variant. The Rc gene exon-2 sequences were similar in the three black rice cultivars. The GT gene sequence was identified as a new variant that correlates with the purple stem, leaf, bran, and whole grain morphology seen exclusively in the BRWJ cultivar. The anthocyanin composition in Java black rice is diverse. The highest cyanidin level was seen in BRWJ and the highest level of peonidin-3-O-glucoside in BREJ. Proteomic profiling of the black rice cultivars demonstrated that the expression of proteins that might be related to the levels of anthocyanin synthesis varied. These studies conclude that the genomic, proteomic and anthocyanins composition of Java black rice cultivars may be used the improvement of their functional nutrition values.

The Caprine casein-alpha-S2 protein modulates the molecular mechanism of insulin signal transduction in type 2 diabetes rat.

This study purpose was to investigate the association of casein-alpha-S2 protein of Caprine milk and molecular mechanismofinsulin signaltransduction in type2 diabetes mellitus (T2DM). The Caprine milk CSN1S2 protein treatment of 0, 375, 750, and 1500mg/kg BW were conducted to the control and T2DM rats. We observed several physiological parameters of all rats. The levels of insulin and TNF-α in the plasma were measured using ELISA.The expressions of proteins and mRNA levels of diabetes-related genes in the pancreas tissues were analyzed using Western Blotting and Real-Time PCR, respectively. Our study found that diabetic rats had lower body weight, food intake, and fecal weight compared with control rats. The Caprine milk CSN1S2 protein consumption affected the body weight of diabetic rats to increase, especially at the dose of 750mg/kg BW.Interestingly, the genes associated with insulin signaling were improved by the CSN1S2 protein treatment in diabetic rats, although their blood glucose and cholesterol level were not affected. The diabetic rats showed an elevated insulin level and GLUT4 protein expression after treatment. We also reported that the CSN1S2-treated diabetic rats had a gradually reduced expression of TNF-α and VCAM-1 in dose-dependent. Moreover, the 750mg/kg BW of CSN1S2 treatment enhanced the mRNA expressions of INS-receptor, GLUT4, IGF-1, CAMKK, and CAMKIV in diabetic rats. The ability of Caprine milk CSN1S2 protein to regulate the molecular mechanisms in the diabetes-signaling pathway indicated its potential therapeutic effects on diabetes management.

Thalassemia Major and Intermedia Patients in East Java do not Show Fetal Hemoglobin Level Difference in Relation to XMNI Polymorphism.

INTRODUCTION: Thalassemia is a genetic disorder, which shows, varies phenotype due to genetic modifier. XmnI is one of the genetic modifiers which affect clinical severity in thalassemia. XmnI polymorphism may increase HbF production beyond fetal life, thus ameliorating the clinical phenotype. AIM: this study aimed to investigate the difference in HbF level and the relation of HbF level and XmnI polymorphism in Thalassemia Major (TM) and Thalassemia Intermedia (TI) patients. METHODS: forty-eight beta thalassemia patients (28 males and 20 females), consists of 16 TM and 32 TI; mean age, 25.30 year old. Hemoglobin Fetal and HbA2 level were determined using High performance Liquid Chromatography (HPLC), and XmnI polymorphism was confirmed by PCR-RFLP. Statistical analysis was done using T-test, Mann-Whitney and Pearson Chi-square. RESULTS: The frequency of heterozygote (+/-) XmnI polymorphism in TM and TI patients was 56.25% vs 71.87%, while the frequency of homozygote (-/-) in TM and TI was 43.75% vs 28.13% with p value >0.05. The insignificant difference also found in HbF level between XmnI +/- and -/- in TM and TI patients. CONCLUSION: This study revealed that thalassemia major and thalassemia intermedia patients in East Java showed similar XmnI polymorphism. These phenomena also showed by HbF level in relation to XmnI polymorphism in the phenotype groups (TM and TI).

An in Silico Approach Reveals the Potential Function of Cyanidin-3-o-glucoside of Red Rice in Inhibiting the Advanced Glycation End Products (AGES)-Receptor (RAGE) Signaling Pathway.

INTRODUCTION: Advanced glycation end products (AGEs) contribute to the pathogenesis of chronic inflammation, diabetes, micro and macrovascular complications, and neurodegenerative diseases through the binding with RAGE. Natural compounds can act as an alternative in disease therapy related to the AGEs-RAGE interactions. Cyanidin-3-O-glucoside is one of the potential anthocyanins found in red rice. Cyanidin-3-O-glucoside in red rice may interfere with the AGEs-RAGE signaling so that the potential mechanism of their interaction needs to be elucidated. AIM: This study aimed to investigate the potency of cyanidin-3-O-glucoside in red rice as an inhibitor of AGE-RAGE signaling pathway through in silico analysis. METHODS: Our study used the 3D structures of AGEs and Cyanidin-3-O-glucoside compounds from PubChem and Receptor for AGEs (RAGE) from the RCSB Protein Data Bank (PDB) database. The molecular interactions of those compounds and RAGE were established using Hex 8.0 software, then visualized using Discovery Studio 2016 software. RESULTS: Argypirimidine, pentosidine, pyrralline, and imidazole bound to the ligand-binding domain of RAGE with the binding energy of -247 kcal/mol, -350.4 kcal/mol, -591.1 kcal/mol, and -100.4 kcal/mol, respectively. The presence of cyanidin-3-O-glucoside in the imidazole-RAGE-cyanidin-3-O-glucoside complex could inhibit the interaction of imidazole-RAGE with a binding energy of -299 kcal/mol, which was lower than of imidazole-RAGE complex. The establishment of AGEs-Cyanidin-3-O-glucoside-RAGE complex showed that cyanidin-3-O-glucoside, which bound first to Argypirimidine and Pyrralline, could bound to RAGE at the same residue as those two AGEs did with the binding energy of -411.8 kcal/mol and -1305 kcal/mol, respectively. Based on the binding site location and energy, cyanidin-3-O-glucoside might have a biological function as an inhibitor of AGEs-RAGE interactions, which was more likely through the establishment of AGEs-cyanidin-3-O-glucoside-RAGE. CONCLUSION: This study suggests that cyanidin-3-O-glucoside in red rice can be a potential AGEs-RAGE inhibitor, leading to the regulation of the pro-inflammatory and oxidative damage in the cellular pathway.

Virtual Prediction of the Delphinidin-3-O-glucoside and Peonidin-3-O-glucoside as Anti-inflammatory of TNF-α Signaling.

INTRODUCTION: Anthocyanin is the bioactive compound in black rice, which promotes some health benefits for human body. Present study revealed that black rice anthocyanins improve the biomarker of the metabolic syndrome, such as tumor necrosis factor alpha (TNF-α). However, the mechanism of anthocyanin in preventing metabolic syndrome has not been elucidated. AIM: This study was performed to identify the interaction of six types of black rice anthocyanin towards TNF-α protein and TNF-α receptor through in silico studies, to assess the molecular properties and bioactivity of black rice anthocyanin. METHODS: We retrieved the black rice anthocyanin compounds from the PubChem database and the proteins (TNF-α protein and TNF-α receptor) from Protein Data Bank (PDB) database. Protein and ligands were docked using Hex 8.0 software and visualized by Discovery Studio 4.1 program. RESULTS: This study found the possibility that black rice anthocyanins interacted with TNF-α have no influence into TNF-α and TNF-α receptor interaction. The binding of delphinidin-3-O-glucoside & peonidin-3-O-glucoside to TNF-α receptor inhibited the TNF-α and TNF-α receptor signaling. The black rice anthocyanins had low activity as a drug. Interestingly, black rice anthocyanins had a potency as an antioxidant due to the hydrogen donor or acceptor in their structure, as protein kinase inhibitor, nuclear receptor ligand, and enzyme kinase inhibitor. CONCLUSION: This study suggests that delphinidin-3-O-glucoside and peonidin-3-O-glucoside might have function as anti-inflammatory factor related with TNF-α signaling.

Effect of Hyperglycemia to The mRNA Level and Protein Expression of Perlecan at Rat Model of Osteoarthritis with Diabetes Mellitus Type 1.

INTRODUCTION: Previous research found that diabetes mellitus capable to aggravate osteoarthritis disease. In brief, the hyperglycemia condition in diabetes mellitus has an impact on protein glycation of all joint components, including molecule, such as perlecan. The protein expression of perlecan reflects the presence amount of perlecan in the matrix of articular cartilage. However, the impact of hyperglycemia on articular perlecan has not been explained. Moreover, the role of perlecan as a mechanotransducer for chondrocytes in type 1 Diabetes mellitus remains unclear. AIM: This research aims to analyze the effect of hyperglycemia in type 1 Diabetes mellitus to the mRNA level and protein expression of perlecan. METHODS: Thirty-five adult male rats were divided into seven groups, such as three groups of rat model with anterior cruciate ligament transection (ACLT) at right knee (ACLT1, ACLT2, ACLT3); three groups of rats with ACLT at right knee which followed by Streptozotocin injection for diabetic mice model (DM1, DM2, DM3); and control group (N). Rat sacrificed at the third week, fourth week, and sixth week after two months of maintenance. The mRNA level and protein expression were analyzed by using PCR and Western blot. All of data was analyzed by ANOVA. RESULTS: Protein expression of perlecan in ACLT mice with diabetes mellitus (DM1, DM2, DM3 group) was gradually decreased according to the increased hyperglycemia duration. Whilst, protein expression of perlecan within ACLT mice without diabetes mellitus (ACLT1, ACLT2, ACLT3 group) was increased. The similar result also demonstrated by the mRNA level of perlecan. Group of DM1, DM2, DM3 exhibited decreased mRNA level of perlecan over the hyperglycemia duration. While, ACLT1, ACLT2, and ACLT3 group had a gradually increased of perlecan mRNA level. CONCLUSION: Hyperglycemia on osteoarthritic condition decreased mRNA level and protein expression of perlecan which increased the severity of osteoarthritis disease.

Cdc2-like kinase 2 (Clk2) promotes early neural development in Xenopus embryos.

Neural induction and patterning in vertebrates are regulated during early development by several morphogens, such as bone morphogenetic proteins (BMPs) and fibroblast growth factors (FGFs). Ventral ectoderm differentiates into epidermis in response to BMPs, whereas BMP signaling is tightly inhibited in the dorsal ectoderm which develops into neural tissues. Here, we show that Cdc2-like kinase 2 (Clk2) promotes early neural development and inhibits epidermis differentiation in Xenopus embryos. clk2 is specifically expressed in neural tissues along the anterior-posterior axis during early Xenopus embryogenesis. When overexpressed in ectodermal explants, Clk2 induces the expression of both anterior and posterior neural marker genes. In agreement with this observation, overexpression of Clk2 in whole embryos expands the neural plate at the expense of epidermal ectoderm. Interestingly, the neural-inducing activity of Clk2 is increased following BMP inhibition and activation of the FGF signaling pathway in ectodermal explants. Clk2 also downregulates the level of p-Smad1/5/8 in cooperation with BMP inhibition, in addition to increasing the level of activated MAPK together with FGF. These results suggest that Clk2 plays a role in early neural development of Xenopus possibly via modulation of morphogen signals such as the BMP and FGF pathways.

Chondrocyte Intracellular Matrix Strain Fields of Articular Cartilage Surface in Hyperglycemia Model of Rat: Cellular Morphological Study.

INTRODUCTION: Chondrocyte is one cell in articular cartilage was products many proteins, molecules, and other factors. The external influence of cartilage, such as: hyperglycemia was entering joint capsule and impact to the chondrocytes and the cartilage. Hyperglycemia caused modification of heparan sulfate proteoglycan 2 (perlecan) proteins through glycation process. AIM: The aim of this study was to analyze morphological changing of chondrocytes pericellular matrix by the influence of hyperglycemia. MATERIAL AND METHODS: Eighteen adult male rats were divided into six groups: control, rat treated with sugar intake was 0.5 mg/kg, 0.75 mg/ kg, 1mg/kg, 1.5 g/kg and 2 mg/kg of body weight. The animal model was dislocated and left knee was taken to observe changing of chondrocytes pericellular matrix strain fields by Scanning Electron Microscope (SEM) from perpendicular to femoral condylus cartilage. RESULTS: Changing of chondrocytes intracellular matrix strain fields as changing of cell diameters and cell distances at group control and group I to group V, which cell diameters was lower level and cell distances was the highest level at over diet 2. This changing of intracellular matrix strain fields was corresponding to changing chondrocytes morphology in hyperglycemia condition, due to hypertrophic stage as adaptive responses. This research as based on next research for accomplish of hyperglycemia influence to morphology articular cartilage changing to prevent degeneration of cartilage towards osteoarthritis. CONCLUSIONS: Present study concludes that hyperglycemia influence to chondrocyte intracellular matrix strain fields changing.

The Level Short Chain Fatty Acids and HSP 70 in Colorectal Cancer and Non-Colorectal Cancer.

INTRODUCTION: Microbial involvement in colorectal cancer (CRC) is now well established. Short Chain Fatty Acids (SCFA) is the main products of anaerobic microbial fermentation in the large intestine and affects colonic health. SCFA mainly produced as microbial metabolites, acetate, propionate, and butyrate acids. Several in vitro studies showed that butyrate induce expression of heat shock protein (HSP) 70 that has function in the beginning of apoptosis. AIM: The aim of this study was investigating the differences level SCFA and HSP 70 expression in CRC compared with non-CRC patients. MATERIAL AND METHODS: The study consists of 14 patients diagnosed with CRC and 14 non-CRC patients. Stool sample were analyzed for SCFA (acetate, propionate, and butyrate acids) with gas chromatography and the result is given as µg/mL and the protein expression of HSP70 was determined by immunohistochemistry (IHC) and haematoxylin-eosin staining to determine the morphological changes in colon tissue. RESULTS: We found that CRC patients had lower level of acetate, propionate and butyrate acids than non-CRC. Whereas in CRC patients, the mean concentration of acetate was 8,55 µg/mL, propionate was 5,61 µg/mL and butyrate acids were 3,79 µg/mL respectively (all P < 0.05). And among the samples of patients with colorectal cancer was obtained the highest expression of HSP-70. CONCLUSIONS: Short chain fatty acids were indirectly contributed in the role of pathogenesis in CRC despite another factor could affect for this disease.

Differential Expression of IL-10 Gene and Protein in Target Tissues of Rattus Norvegicus Strain Wistar Model Type 2 Diabetes Mellitus (T2DM).

INTRODUCTION: Type 2 diabetes mellitus (T2DM) is a chronic metabolic disease caused by insulin resistance. Insulin resistance leads to hyperglycaemia that causes complication such as microangiopathy and macroangiopathy. The immune system of T2DM will be produce IL-10 as an anti-inflammatory cytokine role immune-stimulator and immunosuppressant in the organ system. This present study investigated of IL-10 gene profile and protein expression in the rat organ (Rattus norvegicus) strain Wistar model T2DM. MATERIAL AND METHODS: This research was used three of male rats group T2DM and three of male of normal rat as a control. The DNA tissues were isolated, amplified and sequenced by using IL-10 gene primer. The IL-10 protein profile and expression of rat tissues was analyzed using Experion-Pro260 gel and dot blotting using IL-10 antibody. RESULTS: This study showed the differential expression of IL-10 gene profile among tissues among normal and T2DM groups. The IL-10 gene sequences, we found eight mutations in brain and twenty-seven mutations on gastric of T2DM group compare with control group, meanwhile there are no mutation in other tissues of both groups. The protein profile of all tissues in both groups was completely diverse as proper. Moreover, the level expression of IL-10 of heart, lung, gastric and kidney of T2DM group was lower than other tissues of both groups. CONCLUSION: This study concludes that T2DM animal model triggering mutation of IL-10 gene sequences of brain and gastric and induced the increasing level expression of IL-10 of ileum, brain and liver.