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Background: Obesity is a global epidemic caused by excessive body fat, which is increasing free fatty acids in the liver, causing oxidative stress and liver cell damage. Green tea yogurt (GTY) with encapsulated Lacticaseibacillus paracasei E1 (GTY-LpE1) might have a beneficial effect in reducing liver cell damage. This study was conducted determine GTY-LpE1 effect on superoxide dismutase (SOD) expression, malondialdehyde (MDA) expression and liver histopathology in high-fat high-fructose diet (HFFD) mice. Material and Methods: A completely randomized design (CRD) with 7 groups, including normal diet (ND) group, HFFD group, 1.3 mg/kg BW simvastatin (SIM)-administered HFFD group, 5 g/kg BW probiotic yoghurt (PY)-administered HFFD (PY), 2.5 g/kg BW GTY-administered HFFD (2.5 GTY), 5 g/kg BW GTY-administered HFFD (5 GTY), and 10 g/kg BW GTY-administered HFFD (10 GTY). The diet was given for 16 weeks, followed by oral administration of sim/yoghurt during the last 4 weeks. Mice were sacrificed and the liver was collected. SOD and MDA expression were analyzed by flow cytometry. Histopathology analysis was done by evaluating hematoxylin-eosin (HE) staining of the liver. Result: The percentage of necrotic cells were 34.55, 34.31, and 21.95%, when treated with 2.5, 5, and 10 g/kg BW with GTY-administered HFFD, respectively, these were lower than the ones in the HFFD group (69.49%). The percentage of MDA expression were 15.55, 18.69, and 22.42%, respectively, these were lower than the ones in the HFFD group as well. The percentage of SOD expression were 9.49, 7.85, and 11.11%, respectively, these were higher than the ones in the HFFD group (3.44%). Conclusion: GTY-LpE1 could decrease the number of necrotic cells in the HFFD mice livers and improve the hepatocyte damage by reducing MDA expression and enhancing SOD expression. GTY-LpE1 can be used as an alternative food to control obesity. Keywords: alginate, chitosan, encapsulation, green tea, probiotic

In recent years, the development of probiotic-based fermented products with halal status has been a concern. The use of growth medium has relied on de Man Rogosa and Sharpe (MRS) as a relatively expensive standard medium, and its halal status is still uncertain. Extensive research has been carried out to investigate the development of low-cost halal alternative media for the cultivation of probiotic lactic acid bacteria (LAB). This study aimed to develop a probiotic halal and low-cost culture medium using a tofu whey-based medium. This study used three tofu whey-based media – A (tofu whey 100%), B (tofu whey 94.5%, molasses 3%, skim milk 2.5%), C (tofu whey 92.5%, molasses 3%, cheese whey 2.5%, tomato extract 2%), and MRS broth as a standard medium. Bacterial populations, total sugars utilized, total lactic acids produced, low pH (2.0) tolerance, and high bile salt concentration (oxgall 1.5%) were assayed. The highest bacterial population after 48 h of incubation was shown by medium B compared to medium MRS (12.34 ± 0.87 and 11.48 ± 0.3 log CFU/mL). Total sugars utilized by 48.28 ± 2.89, 38.89 ± 6.94, 39.14 ± 4.24, and 76.00 ± 1.41 %; on the other hand, total lactic acids produced by 0.16 ± 0.12, 0.03 ± 0.04, 0.31 ± 0.03, and 2.25 ± 1.48 % in A, B, C, and MRS, respectively. Probiotic tolerance at low pH and the presence of bile salts of Lactiplantibacillus plantarum D4 consistently showed a high survival rate in medium B compared to MRS. Based on these results, the components and proportions used in medium B were suitable for the growth of L. plantarum D4 as a halal probiotic starter candidate.

Background and Aim: Antimicrobial resistance (AMR) is becoming a public health concern. Foodborne pathogens are infectious agents that can be transmitted from animals to humans through food and can become resistant due to misuse and overuse of antibiotics, especially in poultry. This study aimed to detect the prevalence of multidrug-resistant and extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolated from local and broiler chickens at the Cibinong market, West Java, Indonesia. Materials and Methods: A total of 60 cloacal swab samples from 30 local and broiler chickens sold at the Cibinong market in West Java were obtained by random sampling. From these samples, 39 E. coli isolates were obtained after being cultured on eosin methylene blue agar and molecularly identified using polymerase chain reaction (PCR). Six antibiotic disks were used for the antibiotic sensitivity test against E. coli isolates cultured on Mueller-Hinton agar. PCR was performed to detect ESBL genes (blaTEM, blaSHV, and blaCTX-M). Results: A total of 76.47% (39/51) cloacal swab samples were positive for E. coli. All E. coli isolates were sensitive to imipenem (100%), and 38 isolates were sensitive to cefoxitin (FOX) (97.4%). On average, the isolates were sensitive to amoxicillin-clavulanic acid (AMC) (69.2%) and ceftriaxone (CRO) (89.7%). E. coli isolates were occasionally resistant to enrofloxacin (25.64%), followed by gentamicin (20.51%), CRO (10.25%), AMC (7.69%), and FOX (2.56%). The prevalence of E. coli AMR was 10.25% (4/39). All four multidrug-resistant E. coli isolates (blaTEM and blaCTX-M) were confirmed to have the ESBL gene based on PCR. Conclusion: The prevalence of multidrug-resistant and ESBL-producing E. coli is still found, proving that there is still inappropriate use of antibiotics and a need for strict supervision of their use, especially around Cibinong market, West Java.

Cancer death cases have increased yearly, and there are estimated to be 21.6 million cancer cases in 2030. Studies of herbal compounds for cancer treatment alternatives are essential because cancer treatment is relatively expensive and has adverse effects. Phyllanthus niruri (Pn) and Catharanthus roseus (Cr) are plants that are known as herbal medicines. Combining the two plants is expected to prevent and enhance the immune system in breast cancer cases. This study aims to analyze the anti-cancer and immunomodulatory effects of P. niruri and C. roseus extract (PCE) in modulating macrophage polarization in breast cancer mice. Experimental animals are divided into six groups and there is healthy control (normal mice), cancer (DMBA-induced mice), cancer mice with cisplatin administration, cancer mice with PCE administration with three different doses, including dose 1 (500 mg/kg Pn + 15 mg/kg Cr), dose 2 (1000 mg/kg Pn + 75 mg/kg Cr), and dose 3 (2000 mg/kg Pn + 375 mg/kg Cr). The mice were injected with DMBA once a week for six weeks to induce cancer in mice. The breast cancer mice model was administered with PCE orally for 14 days. The expression of CD11b+IL-10+ and CD11b+IFN-γ+ demonstrated macrophage polarization. The results showed that breast cancer induction using DMBA increased the level of IL-10 and decreased the level of IFN-γ significantly compared to the normal group (p < 0.05). In specific doses, administration of PCE could reduce IL-10 levels and increase the level of IFN-γ significantly (p < 0.05). PCE can modulate the polarization of macrophages by suppressing the M2-like macrophage and increasing the M1-like macrophage. The ability of PCE to modulate macrophage polarization indicates that the combination of P. niruri and C. roseus has activity as an anti-cancer.

Free-range chicken is livestock reared to support the humans' need for protein alongside its ritualistic use in traditional medicine to treat diseases. This study investigates the diversity of bacterial communities in the free-range chicken cecum reared in different East Nusa Tenggara Province localities comprising Sambi 1, Sambi 2 villages, Labuan Bajo, and Kupang City. The extracted chromosomal DNA was subjected to next-generation sequencing using the V3-V4 region primers. Results revealed that the Kupang chicken cecum had the highest total tags, while the Sambi 2 village recorded the lowest. Similarly, Sambi 2 chicken cecum exhibited the highest unique tags (6662) and OTUs number (1261), while the Kupang samples gave the lowest at 2550 and 745, respectively. The Shannon diversity index for bacterial diversity demonstrated that cecum samples from Labuan Bajo (5.679) were more diverse than Sambi 1 (5.378), Sambi 2 (5.653), and Kupang samples (3.77). The bacteria with the highest dominance index (0.935) was found in Sambi 2, while the lowest was observed in the Kupang samples (0.082). The three bacterial phyla showing the highest relative abundance were those from Sambi 1, Sambi 2, and Labuan Bajo cecum samples, comprising Firmicutes, Bacteroidota, and Actinobacteriota.Conversely, the Kupang samples showed an abundance of Firmicutes, Bacteroidota, and Campilobacterota, compared to the Lactobacillus-dominated Kupang, Sambi 1, and Sambi 2 chicken cecum samples. The highest relative abundance for Bifidobacterium occurred in Sambi 1 and Sambi 2 chicken cecum samples, the Kupang samples were Campylobacter dominated, and Olsenella was abundant in the Labuan Bajo samples. Intriguingly, the bacterial composition in the tested chicken cecum samples largely comprised beneficial bacteria such as the lactic acid bacteria group. This bacterial group can be further characterized for obtaining probiotic cultures that could improve the health of free-range chickens.

Probiotic dietary supplements are in demand worldwide due to consumer awareness of healthy diets. However, it is still challenging to develop a functional yogurt by enriching it with a natural product. Green tea (GT) has many positive impacts on health and greater flexibility when mixed in various systems without losing its antioxidant properties. The objective was to evaluate the concentrations of GT and encapsulated Lacticaseibacillus paracasei E1 (LpE1) and its incubation time using Box–Behnken Design (BBD) for developing functional food with better characteristics and rich antioxidants. LpE1 beads were prepared by extrusion, visualized using SEM, and characterized by FT-IR. The BBD was set with three independent variables: GT (2‒4 % w/v), encapsulated LpE1 (2‒4 % w/v), and incubation time (18‒30 h). Our findings demonstrated that LpE1 beads have spherical shapes with 1.89 ± 0.03 mm in size. FTIR demonstrated that chemical interaction between LpE1 and biopolymer functional groups has a minor effect on the LpE1 bead structure. The optimum conditions by RSM with 81 % desirability values were 4 % GT, 2 % encapsulated LpE1, and a 30-h incubation time, resulting in 4.15 pH, 76.28 % WHC, 23.72 % syneresis, 16.49 cP viscosity, 13.67 color (ΔE), 89.84 % DPPH, 380.51 mg GAE/g TPC, and 802.65 mg QE/g TFC. The titratable acidity and viability were 1.39 % and 9.55 log CFU/mL, respectively. A 4 % GT addition reduced the acceptance rate of GT yogurt (GTY) than plain yogurt (PY). Incorporating GTY with encapsulated LpE1 might be promising to improve the physicochemical properties and functional values of yogurt.

Background: The carcinogenic substance 7,12-Dimethylbenz[a]anthracene (DMBA) was commonly used to induce tumor formation in rodents. The development of tumor may trigger higher expression of pro-inflammatory cytokines, which in turn supports tumor progression. In this study, we examined the efficacy of Cyperus rotundus extract (CRE) that was reported to have anti-inflammatory properties. We focused on investigating the levels of activated T lymphocytes and the pro-inflammatory cytokines expressed by macrophages. Methods: Female BALB/c were injected with DMBA subcutaneously. The DMBA exposed mice were given CRE orally in three different doses; 63.33, 158.4, and 316.8 mg/kg. After 14 days, the levels of activated T lymphocytes and pro-inflammatory cytokines were analyzed using flow cytometry. Graphical analysis was done with FlowJo v10 and followed by statistical analysis. Results: The treatment of CRE reduced the population of CD4 and CD8 T cells. The number of activated CD4 and CD8 T cells were also significantly suppressed. The population of macrophages marked by CD11b cells was significantly reduced. Finally, the CRE treatment suppressed the levels of TNF-α, IFN-γ, IL-1β, and IL-6 expressed by macrophages. Conclusion: Our findings suggest that CRE could be a potential agent useful in therapeutic approaches for curing the disease caused by aberrant cells.

Background: The standardization and mechanism of action of Caesalpinia sappan as an anticancer agent are still lacking. This study aimed to understand the mechanism of action of C,sappan extract as an anticancer agent. Methods: This study was conducted using the A549 lung cancer cell line to understand the mechanism of action of C. sappan extract as an anticancer agent. The cytotoxicity activity, cell cycle progression, apoptosis, protein-related apoptosis (i.e., BCL-2and BAX protein) assays, and RNA sequencing were performed level were measured. Moreover, the antioxidant activity, total flavonoids, and phenolics of C.sappan were also assessed. Results: C.sappan has strong antioxidant activity (22.14 ± 0.93 ppm) total flavonoid content of (529.3 ± 4.56 mgQE/g), and phenolics content of (923.37 ± 5 mgGAE/g). The C.sappan ethanol extract inhibited cancer cell growth and arrested at G0/G1 phase of cell cycle, inducing apoptosis by increasing BAX/BCL-2 protein ratio in A549 lung cancer cell line. Furthermore, results from RNA sequencing analysis showed that C.sappan ethanol extract caused downregulation of genes acting on mitochondrial function including adenosine triphosphate (ATP) production and respiration. Conclusions: This study demonstrated that C.sappan has the ability to inhibit cancer cell growth by inducing apoptosis and mitochondrial dysfunction in A549 cells.

Background and purpose: Angiogenesis has been one of the hallmarks of cancer. In recent years, Phyllanthus niruri extract (PNE) was reported to inhibit angiogenesis by decreasing the levels of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1a (HIF-1a) in breast cancer. However, the experimental results were confirmed in cancer cell lines only, whereas the anti-angiogenic activity in animal models has not been demonstrated. In this study, we tried to examine the anti-angiogenic activity of PNE on BALB/c strain mice models that were induced for breast cancer using the carcinogenic substance 7,12- dimethylbenz[a]anthracene (DMBA). Experimental approach: Experimental animals were divided into five different groups; vehicle, DMBA, PNE 500 mg/kg, PNE 1000 mg/kg; and PNE 2000 mg/kg. Mammary carcinogenesis was induced using a subcutaneous injection of 15 mg/kg of DMBA for 12 weeks. Afterward, oral PNE treatment was given for the following 5 weeks. VEGFA and HIF-1a were observed using immunohistochemistry. Endothelial cell markers CD31, CD146, and CD34 were observed using the fluorescent immunohistochemistry method. The levels of interleukin-6 (IL-6), IL-17, and C-X-C motif chemokine (CXCL12) were measured using flow cytometry. Findings/Results: The survival analysis indicated that PNE increased the survival rate of mice (P = 0.043, log-rank test) at all doses. The PNE treatment decreased the immunoreactive score of angiogenic factors (VEGF and HIF-1a), as well as the endothelial cell markers (CD31, CD146, and CD34). The PNE- treated groups also decreased the levels of inflammatory cytokines (IL-6, IL-17, and CXCL12) at all doses. Conclusion and implications: This finding suggests that PNE may inhibit the progression of angiogenesis in breast cancer mice by targeting the hypoxia and inflammatory pathways.

Doktor Mengabdi is a community service program at Universitas Brawijaya that is carried out to improve the welfare of the community. One of the efforts to help the community in completing its packaging is by providing assistance in the form of business equipment. The purpose of this study is to investigate the difference in income of Small medium enterprises, Lele Sehat (IGF) which has been provided with the help of vacuum frying and spinners machines. Data retrieval was done through interviews and live observations. The data were tested for its normality and further analyzed using a paired t-test. The results showed that there was a difference in income of small-medium enterprises difference between before and after getting tool assistance. Prior income was IDR 3,007,000. It was increased to IDR. 4,097,000. This indicates that providing tools to small-medium enterprises will increase income and indirectly improve their welfare.

Breast cancer is the most common type of cancer women suffer from worldwide in 2020 and the 4th leading cause of cancer death. Boesenbergia rotunda is an herb with high potential as an anticancer agent. This study explores the potential bioactive compounds in B. rotunda as anti-breast cancer agents using in silico and in vitro approaches. The in silico study was used for active compound analysis, selection of anticancer compound candidates, prediction of target protein, functional annotation, molecular docking, and molecular dynamics simulation, respectively. The in vitro study was conducted by measurement toxicity, rhodamine 123, and apoptosis assays on T47D cells. Based on the KNApSAcK database, B. rotunda contained 20 metabolites, which are dominated by chalcone and flavonoid groups. Seven of them were predicted to have anticancer activity, namely, sakuranetin, cardamonin, alpinetin, 2S-pinocembrin, 7.4′-dihydroxy-5-methoxyflavanone, 5.6-dehydrokawain, and pinostrobin chalcone. These compounds targeted proteins related to cancer progression pathways such as the PI3K/Akt, FOXO, JAK/STAT, and estrogen signaling pathways. Therefore, these compounds are predicted to inhibit growth and induce apoptosis of cancer cells through their interactions with MMP12, MMP13, CDK4, JAK3, VEGFR1, VEGFR2, and KCNA3. Anticancer activity of B. rotunda through in vitro study confirmed that B. rotunda extract is strong cytotoxic and induces apoptosis of breast cancer cell lines. This study concludes that Boesenbergia rotunda has potency as an anticancer candidate.

Bacteriocin-like inhibitory substances (BLIS) produced by bacteria is a promising future food preservative agent. This study aimed to obtain bacterial strains that can produce broad-spectrum antibacterial agents and identify the best BLIS producer species based on 16S rDNA sequences. The bacterial strains were isolated from fer-mented mare’s milk using MRS and M17 agar medium. The isolates then were initially screened based on its antibacterial activity of crude cells against Staphylococcus aureus ATCC 6538. The selected strains were cultured and harvested for its cell-free supernatant (CFS). The pH of CFS was adjusted to 6.5 then used for antibacterial activity as-says against ten pathogenic bacteria. Also, the proteinaceous nature of BLIS compound was confirmed by testing with proteinase K. The gDNA of selected isolates was extracted and the 16S rDNA was am-plified using the polymerase chain reaction method then sequenced. The 16S rDNA sequences of the selected strains were used to identify the species using BLAST nucleotides from NCBI then the phylogenetic trees were constructed. 32 isolates was obtained, but only three iso-lates (BC9, SB7, and DC12) were selected as a result of antibacterial screening for further assays. The neutralized-CFS (N-CFS) of these isolates exhibited broad-spectrum antibacterial activity. The N-CFS could be assumed as BLIS. The isolate of BC9 was identified as Ba-cillus amyloliquefaciens strain BC9 that has 99.99 % similarity with B. amyloliquefaciens KC-1, SB7 was Lactobacillus plantarum strain SB7 that has 99.99 % similarity with Lb. plantarum JMC 1149T, and DC12 was Lactobacillus rhamnosus strain DC12 that has 100 % sim-ilarity with Lb. rhamnosus DSM 20021T. Thus, the BLIS produced by those strains is potential for future food and beverages preservations.

This study aimed to assess the changing of bacterial density and the physicochemical aspects during natural fermentation of Sumbawa mare’s milk, and to evaluate the dynamics of bacterial population during the natural fermentation using metagenomic approach. Mare’s milk sample obtained from Regency of Dompu were fermented for 60 days. On the day 0, 7, 15, 30 and 60 mare milk sample were collected for further analysis, such as bacterial density enumeration, nutrition content, physical properties of the milk, and total DNA isolation. The total DNA samples obtained were analyzed using next generation sequencing. The density of lactic acid bacteria was decreased along with fermentation periods. Meanwhile, the density of aerobic bacteria on was relatively fluctuated. The physicochemical content of mare’s milk also changed during fermentation periods. Carbohydrate content and total sugar was decrease along with the decreasing of pH value. Moreover, the lipid content increase, and the protein content was fluctuated. The changing in physical properties such as whey color, acidity and gas was observed until the end of mare’s milk natural fermentation process. Using metagenomics analysis, the bacterial diversity from each sample periods categorized as low because of the dominance of Lactobacillus helveticus until the end of the fermentation. Lactobacillus helveticus as a member of LAB did not grow on isolation media on the late stage of fermentation periods (day-60). The presence of uncultivable bacteria can be detected with metagenomic approach, fulfilling the limited information on the bacterial composition of fermented Sumbawa mare’s milk products.

Background: The mangrove, Rhizophora mucronata, an essential source of endophytic bacteria, was investigated for its ability to produce glutaminase-free L-asparaginase. The study aimed to obtain glutaminase-free L-asparaginase-producing endophytic bacteria from the mangrove and to optimize enzyme production. Methods: The screening of L-asparaginase-producing bacteria used modified M9 medium. The potential producer was further analyzed with respect to its species using 16S rRNA gene sequencing. Taguchi experimental design was applied to optimize the enzyme production. Four factors (L-asparagine concentration, pH, temperature, and inoculum concentration) were selected at four levels. Results: The results indicated that the endophytic bacteria Lysinibacillus fusiformis B27 isolated from R. mucronata was a potential producer of glutaminase-free L-asparaginase. The experiment indicated that pH 6, temperature at 35°C, and inoculum concentration of 1.5% enabled the best production and were essential factors. L-asparagine (2%) was less critical for optimum production. Conclusions: L. fusiformis B27, isolated from Rhizophora mucronata, can be optimized for L-ASNase enzyme production using optimization factors (L-ASNase, pH, temperature, and inoculum), which can increase L-ASNase enzyme production by approximately three-fold.

Salmonella Typhi merupakan salah satu bakteri yang menjadi agen penyakit bawaan makanan. Bakteriofage sebagai alternatif penggunaan antibiotika telah digunakan untuk mengendalikan bakteri tersebut. Penelitian ini dilakukan untuk mendapatkan isolat bakteriofage litik yang mampu melisis beberapa bakteri patogen yang diujikan dan mengetahui pengaruh aktivitas bakteriofage litik terhadap pertumbuhan SalmonellaTyphi. Bakteriofage diisolasi dari limbah rumah tangga. Selanjutnya penentuan host range bakteriofage terhadap bakteri patogen lain dilakukan dengan metode spot test. Uji aktivitas bakteriofage terhadap SalmonellaTyphi dilakukan menggunakan metode bacterial challenge test. Berdasarkan hasil isolasi, didapat enam isolat bakteriofage, yaitu B2-St, B3-St, S1-St, S2-St, SL1-St, dan SL3-St. Semua isolat bakteriofage mampu melisiskan sel bakteri Escherichia coli dan Salmonella Typhimurium namun tidak mampu melisiskan Bacillus cereus, Staphylococcus aureus dan Shigella disentriae.Tiga isolat bakteriofagetelah terpilih berdasarkan densitas plaque terbanyak yaitu B2-St, SL3-St dan S2-St. Kemampuan isolat bakteriofage B2-St dalam melisiskan sel Salmonella Typhi lebih tinggi (6,81 ± 0,35 log sel/mL) daripada isolat bakteriofage SL3-St (7,39 ± 0,31 log sel/mL) dan S2-St (7,60 ± 0,27 log sel/mL). Penurunan densitas sel inang terendah oleh ketiga isolat bakteriofage terjadi pada jam ke-4. Bakteriofage B2-St merupakan bakteriofage terbaik dan berpotensi sebagai agen biokontrol Salmonella Typhi.

In this study, the application of an autochthonous microorganism as probiotic on catfish (Clarias sp.) was scarcely reported. This study aimed to obtain probiotic candidates from the digestive tract (intestinal and gastric) of catfish. A total of nine isolates were successfully isolated from the catfish. Almost all bacterial colonies were morphologically round, had flat edges, were yellow, and produced clear zones as a sign of producing acid during culture. The analysis showed that the three isolates had the best activity in inhibiting fish pathogen isolates. Furthermore, molecular analysis revealed that those three isolates were Bacillus velezensis UB-C1, Bacillus amyloliquifaciens UB-C5, and Bacillus cereus UB-C8. Interestingly, those three bacteria were non-lactic acid bacteria.

A wide variety of yeasts have been involved in traditional fermented foods, and potentially contributed to the development of product properties. The existence of indigenous yeasts in dadih, a traditionally fermented form of buffalo milk of West Sumatera, has been reported but an accurate identification is still required to be conducted. This study was aimed to identify yeasts isolated from dadih using a molecular approach, and to evaluate their lactate utilization. A total of 51 isolates were characterized and identified as Pichia jadinii and Candida stellimalicola using PCR amplification of the 5.8S - internal transcribed spacer region combined with restriction fragment length polymorphism (RFLP) analyses and gene sequencing. The former species were the dominant one in this tested product. Their ability to utilize lactate was demonstrated, indicating that they could modify the sensory characteristics of dadih, and hence interact with the indigenous lactic acid bacteria in dadih. The restriction profiles of the dadih yeasts can be used as a data base for rapid identification of yeasts in the future. Further work is still needed to elucidate the dadih yeast ecology.

Background: Turmeric ( Curcuma longa) has high potential as a traditional anticancer drug. This study aimed to analyze the anticancer activity of turmeric ethanol extract on T47D cells and examine the interaction of Akt1 protein with compounds contained in turmeric. Methods: The cytotoxicity assay was conducted using WST-1 reagents. Apoptosis assay used annexin V-PI, whereas cell cycle assay used PI, and then the results were analyzed using a flow cytometer. LC-HRMS analysis was conducted to identify the active compounds. Docking between Akt1 and ligands was performed using Autodock 4.2 software. Molecular dynamics simulations were conducted using YASARA with a time parameter of 20 ns, pH 7.4, and 37°C. Results: The extract had a strong toxicity on T47D cells (cytotoxicity IC 50 value: 26.36 ± 1.55 µg/mL). The extract induced apoptosis of T47D cells at the IC 50 dose (~30% cells) and induced the cell cycle arrest in G1 phase. Curcumin, 2-hydroxycinnamic acid and caryophyllene oxide had lower binding energy into Akt1 than AZD5363 used as a positive control. Curcumin, Ar-turmerone, and α-curcumene bind in the ATP binding pocket of Akt1, so the compounds have a high potential to be an ATP-competitive Akt1 inhibitors. The interaction of Akt1 with the compound contained in turmeric had an RMSD backbone value that was more stable than that of ATP and AZD5363. Root-mean-square fluctuation values indicated that amino acid residues that had an essential role in ligand binding sites were stable during simulation. Conclusions: The turmeric ethanol extract had a potential anti-cancer effect by inducing apoptosis and inhibiting cell cycle progression on T47D cells. The docking analysis showed that the active compounds of the extract, such as curcumin, Ar-turmerone, caryophyllene oxide, and α-curcumene, were able to bind into the ATP binding pocket of Akt1 that might inhibit the protein activity and induce cell cycle arrest.

Background: The standardization and mechanism of action of Caesalpinia sappan as an anticancer agent are still lacking. This study aimed to understand the mechanism of action of C,sappan extract as an anticancer agent. Methods: This study was conducted using the A549 lung cancer cell line to understand the mechanism of action of C. sappan extract as an anticancer agent. The cytotoxicity activity, cell cycle progression, apoptosis, protein-related apoptosis (i.e., BCL-2and BAX protein) assays, and RNA sequencing were performed level were measured. Moreover, the antioxidant activity, total flavonoids, and phenolics of C.sappan were also assessed. Results: C.sappan has strong antioxidant activity (22.14 ± 0.93 ppm) total flavonoid content of (529.3 ± 4.56 mgQE/g), and phenolics content of (923.37 ± 5 mgGAE/g). The C.sappan ethanol extract inhibited cancer cell growth and arrested at G0/G1 phase of cell cycle, inducing apoptosis by increasing BAX/BCL-2 protein ratio in A549 lung cancer cell line. Furthermore, results from RNA sequencing analysis showed that C.sappan ethanol extract caused downregulation of genes acting on mitochondrial function including adenosine triphosphate (ATP) production and respiration. Conclusions: This study demonstrated that C.sappan has the ability to inhibit cancer cell growth by inducing apoptosis and mitochondrial dysfunction in A549 cells.

Background: The mangrove, Rhizophora mucronata, an essential source of endophytic bacteria, was investigated for its ability to produce glutaminase-free L-asparaginase. The study aimed to obtain glutaminase-free L-asparaginase-producing endophytic bacteria from the mangrove and to optimize enzyme production. Methods: The screening of L-asparaginase-producing bacteria used modified M9 medium. The potential producer was further analyzed with respect to its species using 16S rRNA gene sequencing. Taguchi experimental design was applied to optimize the enzyme production. Four factors (L-asparagine concentration, pH, temperature, and inoculum concentration) were selected at four levels. Results: The results indicated that the endophytic bacteria Lysinibacillus fusiformis B27 isolated from R. mucronata was a potential producer of glutaminase-free L-asparaginase. The experiment indicated that pH 6, temperature at 35°C, and inoculum concentration of 1.5% enabled the best production and were essential factors. L-asparagine (2%) was less critical for optimum production. Conclusions: L. fusiformis B27, isolated from Rhizophora mucronata, can be optimized for L-ASNase enzyme production using optimization factors (L-ASNase, pH, temperature, and inoculum), which can increase L-ASNase enzyme production by approximately three-fold.

Aims: Molecular identification of yeast has been conducted on various fermentation products. However, the identification of yeast in fermented Sumbawa mare's milk based on the genotyping method has not been carried out. This study was aimed to determine the diversity profile of yeasts in fermented Sumbawa mare's milk using phenetic characters and PCR-RFLP analysis technique based on the ITS region. Methodology and results: Yeast isolates were phenotypically characterized and visualized in a dendrogram using CLAD97 software. Then, the yeast DNA was extracted using heat treatment and amplified using ITS1 and ITS4 primers. The amplicons were analyzed by RFLP using HindIII and HaeIII enzymes. The phylogenetic tree was constructed using MEGA 7.0. Based on the result of grouping by phenetic analysis and PCR-RFLP, the 12 isolates were divided into four groups with different members. The results of the phenetic analysis were divided into group I (all isolates of Dompu), group II (isolate B3, B4, S3), group III (isolate B5) and group IV (isolate S1). The types of yeast that were identified molecularly and represented each group of PCR-RFLP results included in group I were Kluyveromyces marxianus D1A and K. marxianus D1B, group II: K. marxianus D7, group III: Kazachstania humilis D4, while milk from Bima and Sumbawa has one yeast species as a member of group IV, namely Pichia kudriavzevii B3. Kluyveromyces marxianus was the yeast frequently found in Sumbawa fermented mare's milk. Conclusion, significance and impact of study: Various yeast species as a consortium of the milk samples can contribute to the increasing quality of fermented Sumbawa mare's milk. [ABSTRACT FROM AUTHOR]