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Hepatic fibrosis is an accumulation of extracellular matrix protein due to response of chronic liver disease. CCl4 compound could induce oxidative stress and lead to hepatic fibrosis. Black soybean (Glycine max (L.) Merr.) contained highest isoflavone content than other types of soy. Fermented black soybean tempeh derived from Rhizopus oligosporus will generate higher isoflavone due to anaerobic metabolic reaction and produce fibrinolytic enzyme which could hydrolize fibrin subtrate. This research was aimed to determine the potency of fermented black soybean tempeh extractas antioxidant and hepatoprotector towards GGT-SOD activities and liver histopathology in hepatic fibrosis rats (Rattus norvegicus) induced by CCl4. This research was used 5 control group: negative control group, positive control group induced by CCl4 20% twice weekly for 2 weeks then CCl4 25% twice weekly for 4 weeks with 0.2 mL/100g b.wt. volume, and therapy group number 1, 2 and 3 which was given fermented black soybean tempeh extract with dose 200 mg/kg b.wt., 400 mg/kg b.wt., 800 mg/kg b.wt., respectively, with volume 2 mL. Liver histopathology was observed by Masson Tricrome staining. The research results showed that fermented black soybean tempeh extract could repair the liver damage. It can be concluded that fermented black soybean tempeh extract as a potency as hepatoprotector. [ABSTRACT FROM AUTHOR]

This study aimed to find out the causality of raw materials, food additives and supportive materials used to produce several cooking spice products available in Indonesia and their direct correlation with monosodium glutamate as a flavor enhancing ingredient. Ten brands of cooking spice were randomly selected and observed using Fourier Transform Infrared (FTIR) Spectroscopy. The results of the observation were processed using main component analysis and classification tree. The results of the analysis show that the ten brands of cooking spice are made from similar food additives and supportive materials with eigenvalues of 9.477 or 94.771% of variant. There is a direct causality with synthetic flavor enhancing ingredient of monosodium glutamate and Chinese Flavor Enhancing Ingredient. Such direct causative direct correlation applies to balado flavored cooking spice, currie flavored cooking spice, Indofood magic cooking spice, straw mushroom broth flavored cooking spice, beef broth flavored cooking spice, chicken broth flavored cooking spice, and totole mushroom. The flavored cooking spice has a causative direct correlation with Chinese flavor enhancing ingredient which eigenvalues is of 8.240 or 82.399% of variant. Balado flavored cooking spice, currie flavored cooking spice, Indofood magic cooking spice, mama cooking spice, and totole mushroom flavored cooking spice have a causative direct correlation with monosodium glutamate with eigenvalues of 1.237 or 12.372% of the variant. The results of classification tree analysis indicated that the fixed variable is Chinese flavor enhancing ingredient and the free variables are those 9 (nine) cooking spices (BM08, BM09, BM05, BM04, BM02, BM03, BM10, BM07 and MSG). There are 7 (seven) free variables with a significant value of more than 50%. Those are BM08 (100%), BM09 (96.70%), BM05 (93.30%), BM04 (73.60%), BM02 (68.90%), BM03 (62.10%) and BM10 (57.00%). The fixed variable of MSG indicates that there are 4 (four) free variables with a significant value of more than 50%, namely BM07 (100%), BM10 (73.40%), BM02 (56.90%), and BM04 (54.00%). [ABSTRACT FROM AUTHOR]

Capsaicin is a secondary metabolite of the Chilean plant. In the pharmaceutical field in addition to relieving pain or pain, capsaicin is also known to have anticancer activity because it inhibits certain oncogenic proteins. Screening of components in Capsicum Annum L. against the target proteins AKT1 and MAPK1 is needed as an initial stage of drug discovery. Further screening of Capsaicin compounds for oncogenic proteins produced in Hepatocellular carcinoma (HCC) pathogenesis signaling. In silico data that have been obtained, Capsaicin in chili (Capsicum Annum L.) has a high affinity for MAPK1 and AKT1 receptor/protein targets with energy and potential activity score (Pa) 0.690 for preneoplastic treatment, 0.590 for apoptotic agonists, and 0.366 for antineoplastic activity. Statistical data using Kruskal Wallis obtained information that Capsaicin can inhibit the expression of AKT 1 and MAPK 1 on mice hepatocyte cells induced by AFB1 in vivo administration, therefore it can be a candidate for anticancer drugs. [ABSTRACT FROM AUTHOR]

Background: Type 2 Diabetes Melitus (T2DM) is a condition of insulin resistance that causes extensive tissue damage due to vascular inflammation and oxidative stress. Lipoprotein-associated phospholipase A2 (Lp-PLA2) has anti-inflammatory role as it hydrolyzes atherogenesis mediators such as oxidized LDL (Ox LDL) and platelet activating factor (PAF) but in contrast, it has pro inflammatory effect as it produced lysophosphatidylcholine (lysoPC) and oxidized fatty acid (oxFA). Methods and Results: This study aimed to measure inflammation marker of T2DM in vivo model with Lp-PLA2 selective inhibitor (darapladib) treatment. It used true experimental laboratory and post test only with control group design using 30 spraque dowley rats that is divided into 3 main groups: normal, T2DM, and T2DM with darapladib (DMDP) administration group. Each group are divided into 2 serial treatment times: 8-weeks and 16-weeks intervention. So, there are 6 groups total with 5 rat in each group. Parameter measured was ox-LDL, tumor necrosis factor-α (TNF-α), inducible nitric oxide synthase (iNOS), interleukin 6 (IL-6), PAF, perivascular adipose tissue (PAT) thickness and also blood glucose, lipid profile, and insulin plasma level. ANOVA test result showed that darapladib were significantly (p: 0.000) lowering tissue and blood ox-LDL level, iNOS, PAF, IL-6 and PAT thickness on T2DM in vivo model. Conclusions: Darapladib proved to have anti inflammation role on T2DM model. [ABSTRACT FROM AUTHOR]

The aim of this study is to construct the hGAD65 gene and to identify the hGAD65 clone by using PCR & RFLP. The samples were derived from normal person & DM patient’s blood. Blood DNA was isolated by salting out method and then amplified by PCR with a pair of specific primer, GAD65-F-BamH1-807 & GAD65-R-Xho1-945. The PCR-product was cloned into vector pET-28a and the pET28a-hGAD65-clone was transformed into E.coli BL21 competent cells. The pET28a-hGAD65-clone was confirmed by PCR and RFLP by BamH1 & XhoI. The PCR product of pET28a-hGAD65-clone was one band of 159bp and has two bands 5.3 kb and 159 bp by RFLPwith both restriction enzymes. The GAD65 protein is expressed in 65kD of pET28a-hGAD65-clone. PET28a-hGAD65-clone was able to recognize by gold standard monoclonal antibody specifically. These results indicated that the hGAD65 gene inserted into pET28a properly and provided the GAD65 protein expression. Key words: hGAD65, PCR, pET-28a, RFLP

Biocathode for biofuel cell was prepared by covalently immobilizedLaccaseon CNT (CNT-Laccase) using glutaraldehyde as conjugates. Successful laccase immobilization was confirmed by Fourier Transform Infrared (FTIR) Spectrophotometry, Surface Electron Microscopy (SEM) and Thermogravimetric Analysis (TGA). Immobilization affected Laccase enzymatic activity where it boosts the stability at high temperature and neutral pH. At temperature 65ºC, free Laccase completely loss its activity, while CNT-Laccase still retaining 57.12% of its activity at 45ºC. The activity of CNT-Laccase at pH 7 was 7.04% of activity at pH 5 which was higher than that of free Laccase. CNT-Laccase was able to perform oxygen electroreduction with addition ABTS (2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) as mediator. Performance of oxygen electroreduction activity was also determined by type and composition of binding polymer. Nafion was able to provide better environment for oxygen electroreduction activity compare to polyvinyl alcohol (PVA). Current density resulted in using Nafion in ratio 1:10 to buffer volume was 1.31 mA/cm2, which was higher than that of PVA (1.01 mA/cm2). Increasing binding polymer ratio into 1:2 and 1:1 undermined oxygen electroreduction activity.

The roses known to contain anthocyanins pigments and as a bioactive compound which can act as free radical catcher material. When harvest season arrives in number will be very abundant. Flowers sorting which can decrease the selling price can be processed in the form of natural dyes for food, cosmetics and medicines / herbs. The purpose of this study is to know applications rose to the quality of some industrial products. The experiment was conducted in (fresh, 2 and 4 days), that performed using randomized block design. Phase II is the application of pigment products of red roses on some industrial products (ie food products, herbal and cosmetics), using the design in accordance with their respective target. The results showed that ucts such as fruit juice, yoghurt, jelly, carbonated beverage (food) and body lotion (cosmetic), and can contribute to the natural color at a time serves as a source of bioactive in effervescent tablets.

Pulp breaking on cocoa processing could be conducted with depolymerization of cocoa pulp by enzyme polygalacturonase (PG) endogenous. This research was aimed to determination the optimum condition of depolymerization of cocoa pulp by enzyme polygalacturonase endogenous during incubation. This research was carried out by incubating cocoa pulp for 6 days on various of optimum condition of PG endogenous treatments, that is: temperature were 32.5o C, 37.5o C, 42.5o C, 47.5o C, 52.5o C and initial pH of pulp were 4.1; 4.6; 5.1, respectively. Optimum condition of depolymerization of cocoa pulp was determination by effectiveness test. Effectiveness test result, based on the highest effectiveness index, showed that the optimum condition of depolymerization of cocoa pulp was 47.5o C incubation temperature and 4,6 initial pH of pulp. Those treatment produced 18.29% of pulp liquid wieght total during 5 days incubation with average per day for pectin content, galacturonic acid content, acid total content, and the change of pH were 1.12%, 8.36%, 0.209 meq NaOH/g and 0,19 in cocoa pulp extract was incubated for 6 days, respectively.

Abstract: In this study, soft-shelled turtle (Pelodiscus sinensis) egg white (SSTEW) proteins were digested by thermolysin and the resulting small peptides were further fractionated by reverse phase chromatography. Peptides with angiotensin I-converting enzyme inhibitory (ACEI) activity from these fractions were screened. A lysozyme-derived peptide, IW-11, from the fraction with the most effective ACEI was identified by liquid chromatography–tandem mass spectrometry (LC–MS/MS) and its purified form showed effective ACEI activity in vitro (IC50 =4.39±0.31μM). The Lineweaver–Burk plots indicated that the inhibition towards ACE caused by this peptide is a competitive inhibition. The molecular docking study further revealed that the ACEI activity of IW-11 is mainly attributed to the formation of hydrogen bonds between the N-terminal residue of IW-11 and the S1 pocket (Ala354 and Tyr523) and the S2′ region (His513 and His353) of ACE. Moreover, the digestion parameters were further optimized and the target peptide (82% purity) was readily obtained (15% yield) without any cumbersome purification procedure. Notably, lysozyme C is the most abundant protein in SSTEW, which implies that an efficient production of this ACEI peptide from SSTEW is promising. Biological significance: Inhibition of ACE has proven to be an effective strategy in prevention and treatment of hypertension and related diseases. Unlike typical synthetic ACE inhibitors which exert well described side effects, food-derived peptides with ACE inhibitory activity may be safer alternatives for hypertension treatment. In this study, we comprehensively identified peptides derived from SSTEW digest using a proteomic approach. IW-11, which is derived from lysozyme, the most abundant protein in SSTEW, showed remarkable inhibition towards ACE. This peptide has been demonstrated to have a competitive inhibitory property which is able to bind to ACE active site and found to be a true inhibitor against ACE according to Lineweaver–Burk plots. Using an optimized thermolysin condition, IW-11 can be readily obtained without any complex purification step, which will benefit its further application to prevention or treatment of hypertension. [Copyright &y& Elsevier]

Introduction: Mesenchymal stem cells taken from adipose tissue have a more accessible access advantage, their retrieval techniques are painless and have multipotent properties which can differentiate osteogenically. Therefore, they have bone regeneration ability in vitro and in vivo studies. The use of allogeneic adipose-derived mesenchymal stem cells (ADMSCs) can regenerate damaged tissue with high-quality without fibrous tissue formation, minimal donor morbidity compared to autografts, and hypo-immunogenic responsiveness. The purpose of this study was to analyze the acceleration of the administration of allogeneic ADMSCs from rat and demineralized dentin matrix (DDM) scaffolds to regenerate alveolar bone resorption in the periodontitis model. Methods: Twenty-five Wistar rats were divided into three groups: control, DDM scaffold group and ADMSCs-DDM composite group. Alveolar bone resorption was induced by injection of LPS P.gingivalis for six weeks in the right mandibular molars. Transplantation was done in periodontal pockets for 7 & 14 days. The parameters were assessed by using scanning electron microscope (SEM), micro-computed tomography (µ-CT), expression collagen type I by immunohistochemistry (IHC) staining and trabecular area and osteoblast with hematoxylin staining (HE). Statistical Analysis was done by using Mann Whitney (p <0.05). Results: There was a significant difference in the expression of collagen type I on the 7th and14th days. Where the expression on day 14 showed significant results (p <0.05) in the ADMSC- DDM composite group compared to DDM scaffold and controls groups. The trabecular area and osteoblast cell count of ADMSC-DDM composite on day 14 was significantly higher than the DDM and control groups (p<0.05). Conclusion: This study concludes that the composite of ADMSCs-DDM on periodontal pockets of Wistar rats has the ability in alveolar bone regeneration. [ABSTRACT FROM AUTHOR]

Objective: Proper management of orthodontic treatment in diabetic patients is essential due to the heightened risk of periodontal tissue breakdown associated with hyperglycemia. Cacao bean extracts (CBE) are known to reduce the inflammatory response and increase synthesis and angiogenesis in periodontitis. Therefore, this study aims to examine the effect of CBE on preventing periodontal tissue breakdown in diabetes with orthodontic force., Methods: A total of 25 Wistar rats were divided randomly into 5 groups, including non-diabetes, diabetes, diabetes cacao 125, 250, and 500 mg/kg BW. Diabetic rats were induced with the stratified dose of Streptozotocin, and a 30-g-force from orthodontic device was applied in all groups. Diabetes cacao group was given CBE for 7 days using a gastric probe. GCF samples were used to analyze the eNOS level through the ELISA method. NFκB, Collagen-1, and FGF-2 expression were then assessed using the immunohistochemical method, while the number of fibroblasts and blood vessels was observed using hematoxylin-eosin stained tissue. The data obtained were analyzed with one-way ANOVA and post hoc tests, with p < 0.05., Results: CBE at a dose of 250 mg/kg BW significantly increased eNOS level, Collagen-1, and FGF-2 expression, and the number of fibroblasts and blood vessels in diabetes groups. Meanwhile, the treatment decreased NFκB expression in diabetes groups (p < 0.05)., Conclusion: This study proved that CBE increased periodontal ligament synthesis and angiogenesis and decreased inflammatory response, thereby preventing periodontal tissue breakdown in diabetic rat models with tooth movement., (© 2024 The Authors.)

Background: There is an essential need for cardiomyocyte regeneration among patients with heart failure. Transplantation of dedifferentiated fat (DFAT) cells may lead to an improvement of cardiomyocyte regeneration among heart failure patients. We believe that DFAT cells are promising candidate cell sources for cardiac regeneration. However, the pathway underlying how DFAT cells of the adipose lineage differentiate into mature cardiomyocytes isn't fully understood., Methods: We conducted an experimental laboratory study on isolated DFAT cells from adipose tissue of healthy adults. Then, we treated cells with different concentrations of reversine (10, 20 and 40 nM), and performed RNA extraction and cDNA synthesis. Next, we used a ceiling culture method based on the buoyancy properties of mature lipid-filled adipocytes. Stemness expression (Octamer-binding transcription factor 4 [Oct4], brachyury, Fetal liver kinase 1 [Flk-1]) was quantified by reverse transcription-quantitative (RT-q)PCR, while cardiomyocyte expression (Transcription factor GATA-4 [GATA4] and cardiac troponin T [cTnT]) was quantified by immunocytochemistry., Results: ANOVA with Tukey's post-hoc found that 10 nM reversine increased greater Flk-1 expression compared to the control group (MD: 5.037 + 0.998; p < 0.001), but there were no significant changes among Oct4 (MD: 0.013 + 1.244; p = 0.99) and brachyury expression (MD: 0.157 + 0.084; p = 0.252). Kruskal-Wallis revealed that the expression of GATA4 (1.65 [0.41-1.98] to 0.015 [0.007-0.034]; p = 0.017) reduced significantly from day 7 until day 21 and cTnT (5.07 [6.62-8.91] to 8.22 [6.81-9.40]; p = 0 .001) increased significantly from day 7 until day 21., Conclusions: Reversine could increase the expression of Flk-1, but it was unable to stimulate the expression of Oct4 and brachyury related to cell stemness. An optimal concentration of 10 nM reversine may have the greatest effect on enhancing the differentiation of DFAT cells into mature cardiomyocytes, as indicated by higher cTnT expression between cells., Competing Interests: No competing interests were disclosed., (Copyright: © 2024 Dharmadjati BB et al.)

This study aims to detect Escherichia coli which encodes beta-lactamase Cefotaxime (blaCTX-M) gene from the reproductive tract of Bali cattle with repeat breeder cases. This research was conducted from June to August 2021 using 16 Bali cattle with repeat breeder cases. The reproductive fluids were taken using a plastic sheet gun which was inserted into a Brain Heart Infusion medium, isolated in eosin-methylene blue agar (EMB) and identified using biochemical tests. Antibiotic susceptibility testing of E. coli was carried out using the disc diffusion method. Double-disk approximation test was used to screen the presence of E. coli which produces Extended-spectrum beta-lactamase. The polymerase chain reaction (PCR) method was used to detect the blaCTX-M gene of E. coli and sequences of the blaCTX-M gene were phylogenetically analyzed. The research results obtained three E. coli isolates from 16 reproductive tract fluids of Bali cattle. Antibiotic sensitivity tests showed that 100% of E. coli was resistant to penicillin G and oxytetracycline. 66.66% of E. coli was resistant to cefotaxime (CTX) and gentamicin, and 33.33% of E. coli was resistant to tetracycline. Escherichia coli isolates that were resistant to penicillin and CTX showed positive results in the double-disk approximation test. The results of E. coli detection using PCR showed that three E. coli isolates encoded the blaCTX-M gene located at 370 bp on gel electrophoresis. The results of the phylogenetic analysis showed that E. coli from the reproductive tract of Bali cattle was related to E. coli that encoded blaCTX-M- 14 isolated from humans., Competing Interests: No conflict of interest in this research, (© 2024 Urmia University. All rights reserved.)

Numerous oxidative stresses are detected in patients with diabetic kidney disease, resulting in insulin resistance that damages the pancreas and kidney. Renal podocytes insensitive to insulin lead to decreased nephrin and podocin and increased insulin receptor serine. The authors did an experiment on diabetic rats to examine the effect of DLBS3233 on repairing insulin resistance., Materials and Methods: Thirty adult male Wistar rats were randomly divided into six groups ( n =5 per group): group of nondiabetic rats as a negative control (group 1); untreated diabetic rats (group 2); diabetic rats treated with DLBS3233 4.5 mg/kg BB (group 3); 9 mg/kg BB (group 4); 18 mg/kg BB (group 5); and diabetic rats treated with pioglitazone (group 6). The authors checked Homeostatic Model Assessment for Insulin Resistance to corroborate insulin resistance prior to DLBS3233 administration in diabetic rats. Immunohistochemistry was performed to examine the expression of renal antimalondialdehyde (MDA) antibodies, nephrin, podocin, and insulin receptor serine. The data were analyzed using analysis of variance and the t -test., Result: In the DBLS3233 group, immunohistochemistry showed enhanced expression of renal nephrin and podocin, as well as diminished expression of anti-MDA antibody, along with decreased insulin receptor serine. From statistical analysis, anti-MDA antibodies and insulin receptor serine showed lower expression, whereas the expression of nephrin and podocin were enhanced compared to untreated groups ( P <0.05)., Conclusion: DLBS3233 reduces oxidative stress by decreasing MDA and improves insulin resistance by increasing the expression of renal nephrin and podocin as well as decreasing insulin receptor serine., Competing Interests: All authors declare no conflict of interest., (Copyright © 2023 The Author(s). Published by Wolters Kluwer Health, Inc.)

Saponin-rich sea cucumber extracts have shown antidiabetic effects in a few reports. Although the triterpene glycosides of sea cucumbers are commonly isolated from their Cuvierian tubules, these are absent in Holothuria atra Jaeger. Therefore, this study intended to investigate the saponin profile in the body wall of H. atra , as well as to assess the α-glucosidase inhibitory activity of the H. atra extracts. The chemical profiling of sea cucumber extracts was conducted by UPLC-HRMS analysis. This resulted in the tentative identification of 11 compounds, 7 of which have not been reported in the H. Atra body wall before. Additionally, two triterpene glycosides were purified and their structures were elucidated based on HRMS and NMR data: desholothurin B (1), and a novel epimer, 12-epi-desholothurin B (2). Moreover, the fatty acid profile of the H. atra body wall was investigated by GC-MS. It was found that the Me90 fraction of the H. atra body wall showed the strongest α-glucosidase inhibitory activity (IC 50 value 0.158 ± 0.002 mg/mL), thus making it more potent than acarbose (IC 50 value 2.340 ± 0.044 mg/mL).

Objective:  The objective of this study was to determine the characteristics of demineralized dentin material sponge (DDMS)., Material and Methods:  An observational study was conducted on DDMS and BPCM. Fourier transform infrared (FTIR) test was performed to determine the characterizations of the materials. Scanning electron microscope-electron dispersive X-ray spectroscopy (SEM-EDX) test was performed to observe the elements contained in the materials., Results:  The infrared spectrum of the DDMS and BPCM functional groups showed the same pattern in each variation, and no significant differences were found. According to SEM analysis, the cavities that make up the membrane were spotted on the surface. Besides, according to the SEM-EDX analysis, DDMS contained chlorine, carbon, and calcium, while BPCM contained carbon, oxygen, and sulfur., Conclusion:  DDMS has the potential to be a biomaterial for bone tissue engineering in terms of the characteristics. DDMS had a structure that almost resembles BPCM as seen from the results of the FTIR graph between DDMS and BPCM. The morphological structure of the two materials in the SEM test appeared to have porosity with various sizes., Competing Interests: None declared., (The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/).)

Background: Graves' disease (GD) is an autoimmune disease, and it accounts for major cases of hyperthyroidism. Antibody against thyroid-stimulating hormone receptor/TSHR (TRAb) is responsible for hyperthyroidism and is considered as a diagnostic marker for GD. Therefore, we developed a recombinant protein of human TSHR-169 (hTSHR-169), which was specifically recognized TRAb in the serum of GD patients and then compare the diagnostic performance between ELISA and dot blot of TRAb tests for their ability to diagnose GD., Methods: 20 GD patients and 20 healthy individuals from the Indonesian population were enrolled. TRAb concentration and density were quantified. Comparative analysis was performed using receiver-operating curve (ROC) analysis., Results: For dot blot assay, the minimum concentration to detect TRAb requiring 100 ng of antigen with antiserum diluted at 1:60. For diagnosing GD, the ELISA yielded a higher AUC compared with the dot blot assay (0.95 and 0.85, respectively). Using the recommended cutoff values, the efficiency of both assays was examined by comparing the specificity and sensitivity of the assays to the clinical diagnosis. The ELISA showed 80% and 95%, while the dot blot assay showed 70% and 95% sensitivity and specificity, respectively., Conclusion: Although the dot blot assay exhibited lower performance than the ELISA method, the dot blot assay is a simple and rapid diagnostic assay that is suitable for diagnosing GD in rural areas, in which healthcare facilities sometimes are not accessible., (© 2022 The Authors. Journal of Clinical Laboratory Analysis published by Wiley Periodicals LLC.)

Nephropathy or kidney disease involves the deterioration of kidney functions, causing severe diseases, such as proteinuria, chronic kidney diseases, and kidney failure. Currently, nephropathy that develops into kidney failure is increasing globally, as indicated by the increasing number of patients undergoing hemodialysis. Some developed analytical methods for nephropathy using albumin, creatinine, uric acid, and the urinary albumin-to-creatine ratio biomarkers, including spectrophotometry, turbidimetric immunoassay, and ELISA, have been reported so far, providing good accuracy and precision. However, WHO has established guidelines for developing diagnostic tools that meet several criteria: Affordable, Sensitive, Specific, User-friendly, Rapid and Robust, Equipment-free, Delivered to those who need it. This means that nephropathy detection can be carried out using a simple method compatible with point-of-care that allows independent urine analysis by patients. For this purpose, the use of paper-based analytical devices (PADs) as an alternative platform for the detection of albumin, creatinine, uric acid, and the urinary albumin-to-creatine ratio were reviewed., (© 2022. The Author(s), under exclusive licence to The Japan Society for Analytical Chemistry.)

Background: Diabetes mellitus (DM) is a chronic and costly disease that has become a primary concern worldwide. Type 1 diabetes mellitus is categorized as an autoimmune disease, which results in islet cell apoptosis and insulin-dependent. GAD 65 is known as a potential marker of impaired pancreatic β cell function that appears in the initial phase of type 1 DM and latent autoimmune diabetes in adults (LADA). This study aimed to develop a novel rapid test of anti-GAD 65 autoantibodies in human serum samples., Methods: We have developed a rapid test for anti-GAD65 autoantibodies in this assay based on the reverse-flow immunochromatography method. Human recombinant-protein antigen for GAD 65 was attached as the control line over the nitrocellulose membrane. On the other side, the goat anti-mouse immunoglobulin G (IgG) was coated on the same membrane as a control line. The positive result for GAD 65 was confirmed by a colloidal gold signal on the strip. Our novel assay analyzed 276 healthy subjects and 51 type 1 diabetes individuals serum samples from several ethnicities in Indonesia for this study., Results: Among the 276 healthy samples, 225 samples were identified as positive for anti-GAD 65 autoantibodies, while 51 samples were negative. Interestingly, the positive results for anti-GAD 65 autoantibodies were linear to the decreasing of high-density lipoprotein (HDL) levels and inversely associated with triglyceride levels. A significant correlation with age was observed in all groups. The sensitivity and specificity test proved that this kit has higher accuracy (AUC value = 0.960)., Conclusion: The significant advantages of our rapid test for anti-GAD 65 autoantibodies provide higher sensitivity, specificity, and stability compared to previous commercial kits. Therefore, it could be proposed as the future clinical diagnostic kit for patient management of type 1 DM., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors.)

Background and Aim: Gliricidia sepium is a medium-sized leguminous plant found widely in tropical to subtropical areas. It has been used as a medicinal ingredient and in rodenticides by local communities in both Indonesia and the Philippines. This study aimed to investigate the wound healing effects of an ointment containing G. sepium leaves on inflammatory cells using a rat model. We also determined its effect on the expression of interleukin (IL) 6 and IL-1β., Materials and Methods: We used 16 Wistar male rats aged approximately 2 months and weighing 150-200 g. They were divided into four treatment groups (T1, positive control; T2, negative control; T3, wounds treated with G. sepium from Indonesia; and T4, wounds treated with G. sepium from the Philippines), and the ointment therapies were applied to wounds for 3 days. Hematoxylin and eosin staining was performed to examine the inflammatory cells microscopically. IL-1β and IL-6 expression were observed immunohistochemically., Results: G. sepium leaves significantly (p<0.05) decreased the number of inflammatory cells, and the expression of IL-1β and IL-6 in the group treated with Indonesian G. sepium leaves was higher than that in the group treated with G. sepium leaves from the Philippines. The leaves contain flavonoids, saponins, and tannins, which act as anti-inflammatory agents to enhance the wound healing process., Conclusion: Our findings suggest that G. sepium leaves from both the Philippines and Indonesia possess wound healing properties., (Copyright: © Aulanni’am, et al.)

Although previously large-scale social restrictions were implemented by the Indonesian government, the total number of coronavirus cases is overcome China in the global ranking per July 18th, 2020, implying a higher infection rate among Indonesian residents. The surge of new coronavirus cases started since the loosening of large-scale social restrictions, thereby implicating that public gathering (including religious gathering) evidently increases transmission [1]. It has been reported that Indonesia's coronavirus disease-19 (COVID-19) mortality rate is the second-highest among Southeast Asian Nations, which may be associated with several health determinants, including biochemical factors and health comorbidity [2], [3], [4], [5], [6], [7]. Because people's adherence to control measures is affected by their attitudes, religious perspectives, and practices (ARP) towards COVID-19. Hence, the information regarding Indonesian's ARP towards COVID-19 post-large-scale social restrictions is required. The data were collected via an online questionnaire, including demographic information (7 items), attitude and practice (5 items), and religious perspective and practice (5 items), from July 11 - 18, 2020, collecting a total of 1,345 respondents. Although our data collection did not provide other precautionary measures (e.g., adequate ventilation). It is notable that most of the religious venues are having a close ventilation system. Hence, this may contribute to the propagation of SARS-CoV-2 transmission [8]. Altogether, these data will help in determining non-health-related factors to prevent the spread of COVID-19., Competing Interests: None to declare., (© 2020 The Author(s).)

Background and Aim: Cattle are an important economic asset for the rural community in East Java Province, Indonesia. The study aimed to provide updated data of cattle farm demography, productivity, as well as the role of the absence of a bull and nematodiosis in reduced productivity of beef cattle in an upland rural area of the province., Materials and Methods: The study was conducted in Sukowono village, Bondowoso region. A Census survey was conducted to collect data through interviews with farmers. Further, 102 fecal samples were taken systematically and processed using a double centrifugation method to investigate the endoparasitic nematodiosis in the cattle population. The demographic data, productivity, and nematodiosis were analyzed descriptively. The difference between proportions was analyzed using Chi-square with 95% confidence limit. The associations were described in risk ratio with 95% confidence interval (CI)., Results: The total cattle population was 814 heads; the range of farm size was 1-7 (median: 2) cattle. Female cattle comprised 81.8% (666/814) of the cattle population but, only 5.5% (23/422) farmers kept both bull and mature female cattle. Pregnancy rate was 26.8% (145/542) of mature female cattle. The delayed first calving time appeared in 24.8% (62/250) of heifers and calving interval of >14 months occurred in 83.2% (149/179) of multiparous cows. The prevalence of endoparasitic nematodiosis was 43.1% (44/102, 95%, CI: 38.1-52.1%). Either the absence of the bull or the nematodiosis did not associate with pregnancy rate or calving interval of cows., Conclusion: This study indicates that the productivity of the cattle in the study area was low but may not associate with the absence of a bull or nematodiosis., (Copyright: © Nugroho, et al.)

Background: The prevalence of malocclusion cases in the orthodontic specialist clinic in Airlangga University's Dental Hospital in Surabaya, Indonesia, in 2014-2016 is fairly high, as 55.34% of the occurrences were identified as class II skeletal malocclusion. This type of skeletal malocclusion, which is usually recognized in adults, occurs as a result of variation during growth and development. Lately, there have been many reports on gene polymorphisms of COL1A1 and COL11A1, which are assumed to be associated with class II skeletal malocclusion in Caucasians., Purpose: This study aims to analyze the relationship between single nucleotide polymorphisms (SNPs) of COL1A1 and COL11A1 with class II skeletal malocclusion in Javanese ethnic group patients with mandibular micrognathism., Materials and Methods: The diagnosis of class II skeletal malocclusion was established using the lateral cephalometric radiographs (ANB angle ≥4°) (n=50). DNA was extracted from the patient's peripheral blood. After that, PCR, electrophoresis, and DNA sequencing were conducted on the extracted DNA based on COL1A1 and COL11A1 primers., Results: The SNPs in COL1A1 are c.20980G/A in 27 patients and c.20980G>A in 8 patients, whereas SNPs in COL11A1 are both c.134373C/A and c.134555C/T in 8 patients and both c.[134373A>C] and c.134582G>A in 10 patients. All samples show the deletion (c.[134227delA]) in COL11A1., Conclusion: SNPs in COL1A1 and COL11A1 have been found in class II skeletal malocclusion of Javanese ethnic group patients. Seventy percent of SNPs in COL1A1 occur in rs.2249492, whereas 36% of newly discovered SNPs appear in COL11A1. All samples also have deletion in COL11A1., Competing Interests: The authors report no conflicts of interest in this work., (© 2020 Ardani et al.)

Background We investigated whether 10-gingerol is able to induce oxidative stress in cumulus cells. Methods For the in-vitro research, we used a cumulus cell culture in M199, containing 10-gingerol in various concentrations (0, 12, 16, and 20 µM), and detected oxidative stress through superoxide dismutase (SOD) activity and malondialdehyde (MDA) concentrations, with incubation periods of 24, 48, 72, and 96 h. The obtained results were confirmed by in-silico studies. Results The in-vitro data revealed that SOD activity and MDA concentration increased with increasing incubation periods: SOD activity at 0 µM (1.39 ± 0.24i), 12 µM (16.42 ± 0.35ab), 16 µM (17.28 ± 0.55ab), 20 µM (17.81 ± 0.12a), with a contribution of 71.1%. MDA concentration at 0 µM (17.82 ± 1.39 l), 12 µM (72.99 ± 0.31c), 16 µM (79.77 ± 4.19b), 20 µM (85.07 ± 2.57a), with a contribution of 73.1%. Based on this, the in-silico data uncovered that 10-gingerol induces oxidative stress in cumulus cells by inhibiting HTR1A functions and inactivating GSK3B and AKT-1. Conclusions 10-gingerol induces oxidative stress in cumulus cells through enhancing SOD activity and MDA concentration by inhibiting HTR1A functions and inactivating GSK3B and AKT-1.

Mycobacterium tuberculosis ( M. tuberculosis ) is the causative agent of tuberculosis in human. One of the major M. tuberculosis virulence factors is early secretory antigenic target of 6-kDa (ESAT-6), and EccB 5 protein encoded by eccB 5 is one of its components. EccB 5 protein is a transmembrane protein in ESX-5 system. The aim of this study is to explore the characteristics of wild-type EccB 5 and its mutant form N426I. We expressed the EccB 5 protein by cloning the mutant and wild-type eccB 5 gene in Escherichia coli ( E. coli ). We compared the protein structure of wild type and mutant form of EccB 5 and found changes in structure around Asn426 (loop structure) in wild type and around Ile426 (β-strand) in the mutant. The truncated recombinant protein of EccB 5 was successfully cloned and expressed using plasmid pCold I in E. coli DH5α and E. coli strain Rosetta-gami B (DE3) and purified as a 38.6 kDa protein by using the affinity column. There was no detectable adenosine triphosphatase activity in truncated forms of EccB 5 and its mutant. In conclusion, our study reveals successful cloning and protein expression of truncated form of eccB5 gene of M. tuberculosis . EccB 5 protein in ESX-5 system may be an important membrane component involved in the transport machinery of type VII secretion system, which is essential for growth and virulence., Competing Interests: No competing financial interests exist., (© Siti Kurniawati et al. 2020; Published by Mary Ann Liebert, Inc.)

Background and Aim: Anisakis is a global challenge for a fish product which may lead to a decrease in economic value and consumers' preference. Skipjack ( Katsuwonus pelamis ) in Kupang, Nusa Tenggara Timur, Indonesia, have important economic value for local fisheries. Anisakis typica is one of the Anisakis species which potent to induce an allergic reaction. However, the study about A. typica involved in the dendritic cells (DCs), T helper 1 (Th 1 ), T helper 2 (Th 2 ), and regulatory T cells (Tregs) is still limited. This study aimed to analyze the dynamic changed of the immune system including DCs, CD4 + T cells, and Tregs after 1 week of A. typica sensitization., Materials and Methods: Twenty-four male Balb/C mice were randomly divided into four groups (n=6), mice treated with crude A. typica extract (CAE) 50, 75, and 100 mg/kg BW, respectively. CAE was given orally per day for a week. At the end of the experiment, the animals were sacrificed and the spleen was collected. DCs were labeled as CD11c + interleukin-6 + (IL-6 + ); CD4 + T cells were distinguished as Th 1 (CD4 + interferon-γ + [IFN-γ + ]) and Th 2 (CD4 + IL-4 + and CD4 + IL-5 + ); Tregs were labeled as CD4 + CD25 + CD62L + . The expression of each cell was determined by flow cytometry., Results: Our result described that CAE elicits CD11c + IL-6 + , CD4 + IFN-γ + , CD4 + IL-4 + , and CD4 + IL-5 + and reduces CD4 + CD25 + CD62L + significantly (p<0.05) in dose-dependent manner in mice after A. typica infection., Conclusion: The Th 1 /Th 2 ratio after A. typica crude extract treatment exhibits a mixed pattern rather than the classical model allergy to food antigens. Our study is expected as a basic understanding of the changes in immune response after A. typic a infection., (Copyright: © Haryadi, et al.)

Purpose: Proprotein convertase subtilisin/kexin type 4 (PCSK4, a 54-kDa protease) is expressed in the plasma membrane of the acrosome in human spermatozoa. It plays a critical role in penetrating the zona pellucida. Synthesis of human anti-PCSK4 might be important for novel male immunocontraception., Materials and Methods: We used semen from adult males as the source of antigen (acrosomal PCSK4). Isolation and antigen characterization were done by immunohistochemistry followed by electrophoresis. Purification of PCSK4 was done by the electroelution method, followed by immunization with an animal model ( Oryctolagus cuniculus ). Antibody was collected, purified, and tested by using Western blot and dot blot. Antibody in vitro potential testing was performed on human spermatozoa by laser scanning microscopy with rhodamine stain under a light microscope and on rat spermatozoa., Results: Human anti-PCSK4 bound with PCSK4 on the head of human spermatozoa and could interfere with rat spermatozoa activity to penetrate the oocyte., Conclusions: The result of this study can be used as a basis to develop new immunocontraceptives targeted towards males. This study shows that antibodies from induction of PCSK4 from spermatozoa may hinder fertilization., Competing Interests: CONFLICTS OF INTEREST: The authors have nothing to disclose.

Background and Aim: Sepsis is characterized by loss of control of the inflammatory response, which can be triggered by various microorganisms and toxic secretions. The mortality rate increases due to impaired endothelial function caused dysfunctional organ systems. Diabetes is closely related to sepsis. The study aimed to determine the method of using animal models of sepsis diabetes through a combination of streptozotocin (STZ) and Staphylococcus aureus infection based on biological marker parameters., Materials and Methods: A total of 30 male Wistar rats of 2.5-3 months old weighing approximately 150-250 g body weight (BW) divided into six treatment groups with five replications per group were used in the study. Treatment A was negative control (healthy rats) and Treatment B was the positive control (with diabetes) where rats were given STZ dose at 45 mg/kg BW on day 8 intraperitoneally (IP). The blood glucose was measured on day 10, Treatment C was a positive control (bacteria), rats inoculated with S. aureus with a concentration of 10 8 CFU/mL on day 8 given IP and observed sepsis conditions on day 10 th . Treatment group (D, E, and F): Rats given STZ dose at 45 mg/kg BW on day 8 th by IP and measured blood glucose on day 10 th , then inoculated with S. aureus with different concentrations of 10 5 CFU/mL, 10 6 CFU/mL, and 10 7 CFU/mL on the 10 th day, respectively, and were later observed the condition of sepsis on day 12 th . Data on diabetes bacteremia were quantitative used blood glucose levels, the bacterial count, and C-reactive protein (CRP) and were analyzed using the one-way analysis of variance test with a confidence level of 95%. Physical examination (temperature and respiration) is qualitative., Results: Physical examination showed that all treatments had a normal temperature, an increased pulse in Groups D, E, and F and a decrease in respiratory rate in the treatment of E and F, the bacteria found in the vital organs in all groups, and CRP levels were not significantly different at all., Conclusion: Animal model of diabetes sepsis can be observed through a combination of pancreas damage, and respiration, the bacteria in the vital organs.

Background: Germline and somatic polymorphisms and mutations of the Androgen Receptor (AR) gene are known to be associated with the incidence of prostate cancer (PCa) in different populations. In this study we assessed germline AR polymorphisms and mutations in PCa patients with prediction of pathogenicity of the identified mutations by in silico analysis. Methods: Diagnosis of PCa was based on histopathology of prostate tissue (Gleason Score criteria) and serum prostate-specific antigen (PSA) levels. Genomic DNA was extracted from peripheral blood of 38 patients. All exons and exon-intron boundaries of AR were amplified using polymerase chain reactions (PCR) followed by Sanger sequencing. In silico analysis was performed using Polyphen-2 and Mutation Taster®. Results: Two polymorphisms, CAG repeat sequence (13-34 repeats in length) and p.Pro214Glu (MAF: 0.0789) located in exon 1 were identified. A missense mutation (c.47C>A/p.Pro146Glu) and in-frame deletion of a CAG sequence leading to loss of Arginine at codon 85 (c.252&#95;254delCAG/p.Arg85-) were identified in a 70 year old patient with a Gleason Score and PSA level of 2 and 2.4ng/dL, respectively. His PSA level decreased to < 0.5 ng/dL after 9 months of androgen deprivation therapy. Identified mutations were predicted to be non-disease causing by Polyphen-2 and Mutation Taster®. Conclusion: Our data demonstrated that the frequency of germline mutations of AR was low in PCa patients in Indonesia (5.26%: 2/38 alleles), so that they are not likely to be major etiological factors. The in silico analysis of identified AR mutations in this study corroborated the clinopathology features of the patient., (Creative Commons Attribution License)

Background: Mycobacterium tuberculosis Complex (MTBC) is a group of Mycobacterium that causes tuberculosis (TB). TB is an infectious disease that remains a global health problem. Indonesia is one of the five countries in the world where TB is the most prevalent and became the country with tle second largest rate of TB in 2014 and 2015. MTBC has high pathogenicity that can cause infections in animals and humans. The most common route of transmission is via airborne droplet nuclei and contact with animals or humans infected with TB. MTBC has many virulence factors. One of these factors is EccB 5 that is encoded by eccB 5 gene. EccB5 is a transmembrane protein-conserved membrane protein and could play a role in inducing damage in host cells, macrophage infection, and may correlate with active disease. The characterization of eccB 5 gene needs to be studied to determine the nucleotide sequences, which may be associated with active disease. The aim of this research was to analyze the nuclotide sequences of eccB 5 gene of MTBC from suspected pulmonary tuberculosis patients, SNPs of eccB 5 gene and possible correlation with the disease, especially in Indonesia., Materials and Methods: Samples were collected from the Tuberculosis Laboratory, Clinical Microbiology of Dr. Soetomo Hospital Surabaya Indonesia. DNA extraction used boiling extraction method and continued nucleic acid amplification using PCR techniques. Primer pairs used eccB 5 SK.. The positivity of DNA specific revealed amplicon in 1592 bp. PCR product was sequenced by 1st Base (First BASE Laboratories Sdn Bhd, Selangor, Malaysia). The sequence analysis used Genetyx-Win version 10.0 (Genetyx Corporation, Tokyo, Japan)., Results: Total isolates of Mycobacterium spp. were 28 and those that showed positive MTBC were 24 isolates and 4 nontuberculosis mycobacteria (NTM) using immunochromatographic test (ICT). The amount of homology from MTBC using blast NCBI was 99%-100%. Two SNPs were found in position c.1277 which revealed replacement of amino acid in 426 of codon position., Conclusion: The sequence of eccB 5 gene of MTBC showed high significant homology, while proposed non-synoymous single nucleotide polymorphisms (nsSNP) may associated with clinical outcomes., Competing Interests: Conflict of interest: No conflict of interest exists concerning this research.

Aim: To investigate the mechanism of pericyte migration through Angiopoietin-2 (Ang-2)/Tie-2 signaling pathway., Methods: We divided the rats into 5 groups. Each diabetic rat model groups injected with Tie-2 inhibitor, ERK1/2 inhibitor, Akt/PKB inhibitor, and DMSO intravitreal. Retinal digest preparation was done to examine the retinal vasculature including pericyte: endothelial ratio, and morphology of pericyte migration. Tie-2, ERK1/2 and Akt/PKB phosporylation were analyzed by confocal laser scanning microscopy., Results: There was a correlation between pericyte migration with increasing Ang-2 ( P <0.05). Pericyte number reduced by 40% (1:2.4) after 5wk diabetes on diabetic rats. The pericyte: endothelial ratio on group with Tie-2 inhibitor were 1:1.8. The same result shows on group with Akt/PKB inhibition. ERK1/2 inhibitor group shows the best results of pericyte: endothelial ratio (1:1.7). Inhibition on Tie-2 receptor decreased the phosphorylation activity of Tie-2, ERK1/2 and Akt/PKB pathway. ERK1/2 inhibition also decreasing the phosphorylation of Tie-2 and Akt/PKB. But on Akt/PKB inhibition, the phosphorylation of Tie-2 and ERK1/2 were relative the same., Conclusion: Ang-2 has a role for pericyte migration on diabetic rats through Tie-2 receptor, ERK1/2 and Akt/PKB pathways. ERK1/2 is a dominant pathway based on the ability to supress another pathway activity and decreasing pericyte migration on diabetic rats.

Objectives: Cumulus cells play a crucial role as essential mediators in the maturation of ova. Ginger contains 10-gingerol, which induces apoptosis in colon cancer cells. Based on this hypothesis, this study aimed to determine whether 10-gingerol is able to induce apoptosis in normal cells, namely, cumulus cells., Methods: This study used an in vitro analysis by culturing Cumulus cells in M199 containing 10-gingerol in various concentrations (12, 16, and 20 μM) and later detected early apoptotic activity using an Annexin V-FITC detection kit., Result: The in vitro data revealed that the number of apoptosis cells increased along with the period of incubation as follows: 12 μM (63.71% ± 2.192%); 16 μM (74.51% ± 4.596%); and 20 μM (78.795% ± 1.435%). The substance 10-gingerol induces apoptosis in cumulus cells by inhibiting HTR1A functions and inactivating GSK3B and AKT-1., Conclusions: These findings indicate that further examination is warranted for 10-gingerol as a contraception agent.

Sperm can change physiology and structure during storage in refrigerator temperature or frozen temperature that caused by cold shock or free radical. The aim of this study to evaluate ultrastructure and fertilizing ability of Limousin bull sperm after storage in cauda epididymal plasma-based (CEP-2) extender with or without 20% egg yolk concentration at refrigerator temperature. Semen sample collected from three Limousin bull were diluted with CEP-2 with 20% egg yolk and CEP-2 without egg yolk, cooled and stored at 4-5 degrees C during eight days. Sperm ultrastructure were observed with scanning electron microscopy (SEM). Fertilizing ability of Limousin bull sperm were assessed on cleavage rate of embryo using in vitro fertilization method. The percentage data were transformed into arcsine before being analysis with ANOVA and Duncan's multiple comparison test. The result of study showed morphologically normal sperm after storage in CEP-2 with 20% egg yolk, whereas in CEP-2 without egg yolk morphologically abnormal sperm especially neck was fractured and head was destroyed. Fertilizing ability of Limousin bull sperm were significantly higher in CEP-2 extender with egg yolk 20% (74.29 +/- 4.95%; p < 0.05) than without egg yolk (30.00 +/- 12.02%; p < 0.05). Egg yolk 20% in CEP-2 extender protected ultrastructure and fertilizing ability after storage during eight days.