Diabetes mellitus is a chronic metabolic disorder due to insulin function insufficiency. This study aimed to test the effectiveness of flower extract of roselle (Hibiscus sabdariffa L.) extract in producing antidiabetic compounds. The inhibition of roselle flower extract on the alpha- glucosidase enzyme was carried out in vitro. Molecular docking was also carried out to bind ligands derived from roselle flower extract's secondary metabolites to the alpha-glucosidase and alpha-amylase enzymes. Based on molecular docking, models have negative binding energies suggesting those ligands make a complex to the site receptor. Kaemferol-3-O-rutinoside and tiliroside become the most stable complex based on the lowest energy score of –9.5 and –8.1 kcal/mol for alpha-amylase and alpha-glucosidase, respectively. The highest antidiabetic activity was obtained at a 100 ppm roselle flower ethanol extract and distilled water with an inhibition value of 100.00 and 99.25%, respectively. The alpha-amylase inhibiting test, using a concentration of 2.5 mg/mL, had an inhibitory activity of 41.77%. The in vitro assessment was conducted using the meta-analysis. The meta-analysis study showed that roselle flower extract could reduce glucose levels in fasting rats better than negative controls (diabetic rats) by 61% than those not given the roselle flower extract.

Honey acts as an antibacterial without side effects, and also contains antiseptic substances which function to inhibit bacterial growth. This study aimed to isolate the Lactic Acid Bacteria (LAB) in Sumbawa white honey and the bioactive compounds produced as pathogenic antibacteria. The 1st stage in this study was the isolation of lactic acid bacteria (LAB) in Sumbawa white honey, then continued with a grading test, morphological test, catalase test, methyl red test, and the last test, namely the antimicrobial test against 5 pathogenic bacteria (Salmonellatyhposa, Staphylococcus aureus, Escherichia coli, Enterobacter ludwigii, and Leclerciaadecarboxylata). Data analysis was performed using the Analysis of variance (ANOVA) test at a confidence level of 0.05 with SPSS 24. Based on the results of sequencing analysis, it was found that the 5 selected isolates were Enterococcus faecium species. The Enterococcus faecium species obtained from the sequencing results had different strains. The accession numbers of the 5 Enterococcus faecium were: Isolate-03 with a percentage of 97.29 % (accession number: KU324920.1), Isolate-07 has a percent identity of 97.36 % (accession number: MF108201.1), Isolate-09 of 97.73 % (accession number: CP041261.3), Isolate-20 with a percentage of 96.40 % (accession number: MN511819.1), and Isolate-24 with a percentage of 98.61 % (accession number: KM495938.1). These isolates can inhibit the growth of all tested pathogenic bacteria treated with 100 % LAB metabolites and were not significantly different (p > 0.05) compared to a positive control (Ampicillin). HIGHLIGHTS Antibacterial compound of Lactic Acid Bacteria (LAB) from Sumbawa white honey Lactic acid bacteria isolation, characterization, and biosprosprection against pathogens Identified LAB by 16s rRNA sequencing gives five strains of Enterococcus faecium All identified LAB metabolites can inhibit all pathogens by similar inhibition percentage with Ampicillin GRAPHICAL ABSTRACT

Type I Interferons (IFNα) are known for their role as biological anticancer agents owing to their cell-apoptosis inducing properties. Development of an appropriate, cost-effective host expression system is crucial for meeting the increasing demand for proteins. Therefore, this study aims to develop codon-optimized IFNα-2b in L. lactis NZ3900. These cells express extracellular protein using the NICE system and Usp 45 signal peptide. To validate the mature form of the expressed protein, the recombinant IFNα-2b was screened in a human colorectal cancer cell line using the cytotoxicity assay. The IFNα-2b was successfully cloned into the pNZ8148 vector, thereby generating recombinant L. lactis pNZ8148-SP Usp45 -IFNα-2b. The computational analysis of codon-optimized IFNα-2b revealed no mutation and amino acid changes; additionally, the codon-optimized IFNα-2b showed 100% similarity with native human IFNα-2b, in the BLAST analysis. The partial size exclusion chromatography (SEC) of extracellular protein yielded a 19 kDa protein, which was further confirmed by its positive binding to anti-IFNα-2b in the western blot analysis. The crude protein and SEC-purified partial fraction showed IC 50 values of 33.22 μg/ml and 127.2 μg/ml, respectively, which indicated better activity than the metabolites of L. lactis NZ3900 (231.8 μg/ml). These values were also comparable with those of the regular anticancer drug tamoxifen (105.5 μg/ml). These results demonstrated L. lactis as a promising host system that functions by utilizing the pNZ8148 NICE system. Meanwhile, codonoptimized usage of the inserted gene increased the optimal protein expression levels, which could be beneficial for its large-scale production. Taken together, the recombinant L. lactis IFNα-2b is a potential alternative treatment for colorectal cancer. Furthermore, its activity was analyzed in the WiDr cell line, to assess its colorectal anticancer activities in vivo.

Background A major discovery in human etiology recognized that cervical cancer is a consequence of an infection caused by some mucosatropic types of human papillomavirus (HPV). Since L1 protein of HPV is able to induce the formation of neutralizing antibodies, it becomes a protein target to develop HPV vaccines. Therefore, this study aims to obtain and analyze the expression of HPV subunit recombinant protein, namely L1 HPV 52 in E. coli BL21 DE3. The raw material used was L1 HPV 52 protein, while the synthetic gene, which is measured at 1473 bp in pD451-MR plasmid, was codon-optimized (ATUM) and successfully integrated into 5643 base pairs (bps) of pETSUMO. Bioinformatic studies were also conducted to analyze B cell epitope, T cell epitope, and immunogenicity prediction for L1HPV52 protein. Results The pETSUMO-L1HPV52 construct was successfully obtained in a correct ligation size when it was cut with EcoRI. Digestion by EcoRI revealed a size of 5953 and 1160 bps for both TA cloning petSUMO vector and gene of interest, respectively. Furthermore, the right direction of construct pETSUMO-L1HPV52 was proven by PCR techniques using specific primer pairs then followed by sequencing, which shows 147 base pairs. Characterization of L1 HPV 52 by SDS-PAGE analysis confirms the presence of a protein band at a size of ~55 kDa with 6.12 mg/L of total protein concentration. Observation under by transmission electron microscope demonstrates the formation of VLP-L1 at a size between 30 and 40 nm in assembly buffer under the condition of pH 5.4. Based on bioinformatics studies, we found that there are three B cell epitopes (GFPDTSFYNPET, DYLQMASEPY, KEKFSADLDQFP) and four T cell epitopes (YLQMASEPY, PYGDSLFFF, DSLFFFLRR, MFVRHFFNR). Moreover, an immunogenicity study shows that among all the T cell epitopes, the one that has the highest affinity value is DSLFFFLRR for Indonesian HLAs. Conclusion Regarding the achievement on successful formation of L1 HPV52-VLPs, followed by some possibilities found from bioinformatics studies, this study suggests promising results for future development of L1 HPV type 52 vaccine in Indonesia.

Background: Hepatitis B is a liver inflammation caused by virus infection leading to acute and chronic conditions. Antigenic compound of HBcAg could induce specific T and B cell that generating high immunity response rather than HBsAg. Therapeutic agent for hepatitis and cancer treatment which has been approved by USFDA is Interferon α-2b (IFNα-2b). This type of class I cytokine plays a role in inhibiting viral replication and modulating adaptive immune system. Objectives: In this study, we analyzed immunogenicity of recombinant HBcAg and IFNα-2b expressed in L. lactis NZ3900. Methods: In vivo test was carried out using female Balb/c mice. Antibody responses for oral immunization were quantified as total IgG using ELISA on days 21, 35 and 51. Safety compound of this oral vaccine candidate was also described by liver and spleen condition after immunization. The differential leukocyte counting was performed to confirm the inflammatory process. Results: Post immunization with L. lactis recombinant strains could induce optimum total humoral immune responses on day 35, with IgG concentration at 4.96±1.03 mg/mL. Single treatment with HBcAg was more potent in inducing immune response (IgG) rather than HBcAg-IFNa-2b combination. Immunization for 51 days could not alleviate animal body weight in each group. The maintaining IgG production until 51 days was just because lymphocytes activities persisted above 70%. The lymphocytes number which achieved 76.3% (P2) and 78.3% (P3) compared to Control group with 62%. Conclusion: Single treatment with recombinant HBcAg expressed in L. lactis NZ3900 was better inducing IgG production and maintaining for 51 days. This result suggested, L. lactis recombinant strain can be as potential vaccine candidate to induce immune response protecting from hepatitis B virus. Moreover, no organ damage was found in liver and spleen of Balb/c mice.