In Silico Screening and Molecular Dynamics Simulation of Potential Anti-Malarial Agents from Zingiberaceae as Potential Plasmodium falciparum Lactate Dehydrogenase (PfLDH) Enzyme Inhibitors.

Malaria continues to be a major public health issue in a number of countries, particularly in tropical regions-the emergence of drug-resistant Plasmodium falciparum encourages new drug discovery research. The key to Plasmodium falciparum survival is energy production up to 100 times greater than other parasites, primarily via the PfLDH. This study targets PfLDH with natural bioactive compounds from the Zingiberaceae family through molecular docking and molecular dynamic studies. Sulcanal, quercetin, shogosulfonic acid C, galanal A and naringenin are the Top 5 compounds with a lower binding energy value than chloroquine, which was used as a control in this study. By binding to NADH and substrate binding site residues, the majority of them are expected to inhibit pyruvate conversion to lactate and NAD+ regeneration. When compared to sulcanal and control drugs, the molecular dynamics (MD) simulation study indicated that quercetin may be the most stable molecule when interacting with PfLDH.

Concatenation of molecular docking and dynamics simulation of human papillomavirus type 16 E7 oncoprotein targeted ligands: In quest of cervical cancer's treatment.

The Human papillomaviruses type 16 E7 oncoprotein is a 98-amino-acid, 11-kilodalton acidic oncoprotein with three conserved portions. Due to its interaction with the pRb-E2F complex, CKII, CKI (mostly p21), and even HDAC1, it possesses strong transformative and carcinogenic qualities that inhibit normal differentiation and cell cycle regulation. Here, we target the E7 oncoprotein using two prior research active compounds: asarinin and thiazolo[3,2-a]benzimidazole-3(2H)-one,2-(2-fluorobenzylideno)-7,8-dimethyl (thiazolo), and valproic acid as a control. We are performing molecular docking followed by molecular dynamic analysis. By acting as competitive inhibitors in the binding site, it was hypothesized that both drugs would inhibit E7-mediated pRb degradation and E7-mediated p21 degradation, resulting in decreased cell cycle progression, immortalization, and proliferation. In addition, we expect that the direct inhibitory action of valproic acid in E7 will target the CKII-mediated phosphorylation pathway necessary for destabilizing p130 and pRb. According to the results of the dynamic simulation, stable interactions exist between every compound. Despite the instability of E7 protein, stability results indicate that both natural chemicals are preferable, with thiazolo outperforming valproic acid.

Understanding hypocholesterolemic activity of soy isoflavones: Completing the puzzle through computational simulations.

The hypocholesterolemic activity of soy isoflavones has been studied, but the exact mechanism underlying the activity remains unclear. This study reveals the proposed mechanism of the cholesterol-lowering effect of soy isoflavones by computational simulations. Daidzin, Glycitin, Genistin, Daidzein, Glycitein, Genistein, Glyceollin I, Glyceollin II, and Glyceollin III were selected to be analyzed their interaction with 3-Hydroxy-3-Methyl-Glutaryl-Coenzyme A Reductase (HMGCR) and Sterol Regulatory Element-Binding Protein 2 (SREBP2) as key factors in cholesterol biosynthesis as well as Proprotein Convertase Subtilisin/Kexin type 9 (PCSK9) as a common target for hypercholesterolemia. Protein-isoflavones interaction was analyzed using AutoDock. According to binding energy calculations, a total of five out of those nine isoflavones, including Glycitin, Genistin, Genistein, Glyceollin II, and Glyceollin III, were favored to be a HMGCR inhibitor but not with SREBP2 and PCSK9. Those isoflavones were then compared with Simvastatin as known inhibitor of HMGCR. Isoflavone with binding energy lower than Simvastatin then directed to molecular dynamics using YASARA and headed into toxicity estimations. Almost all of those isoflavones could bind with HMGCR with better stability than Simvastatin according to molecular dynamics simulations. Toxicity prediction filtered two out of the five isoflavones mentioned earlier as the proper candidate to be an HMGCR inhibitor. Those isoflavones were Genistin and Genistein. In summary, the hypocholesterolemic activity of soy isoflavones may occur by blocking the cholesterol biosynthesis pathway.Communicated by Ramaswamy H. Sarma.

The combination of Elephantopus scaber and Phaleria macrocarpa leaves extract promotes anticancer activity via downregulation of ER-α, Nrf2 and PI3K/AKT/mTOR pathway.

BACKGROUND: Elephantopus scaber and Phaleria macrocarpa have recently been interested as novel anticancer agents. However, there was no scientific evaluation of the anticancer effect of both plant combinations. OBJECTIVE: This study investigated the potential anticancer effects of combined E. scaber and P. macrocarpa leaves extract on human breast cancer cells lines. MATERIALS AND METHODS: T47D cells were treated with the combination of E. scaber and each part of P. macrocarpa (leaves/EL, mesocarp/EM, seed/ES and pericarp/EP). T47D cells were then exposed to three ratios (1:1, 2:1, and 1:2) of the best combination for 24, 48, and 72 h. The cell viability of T47D and TIG-1 cells was assessed using WST-1 assay. The apoptotic hallmarks were determined using FITC Annexin V-PI staining and DNA fragmentation assay. The cell proliferation and cell cycle profiles were analyzed using CFSE (carboxyfluorescein succinimidyl ester) and Propidium iodide-flowcytometry assays. The relative number of p-ERα, p-Nrf2, p-PI3K, p-AKT, and p-mTOR were assessed using flow cytometry. The molecular docking analysis was also performed to confirm the mechanism of the extract in silico. RESULTS: The combination of E. scaber and P. macrocarpa leaves (EL) possessed strong cytotoxic activity (p < 0.05) than other combination groups and cisplatin. EL showed selective killing only to T47D cells. EL at a ratio of 2:1 potentially suppressed the cell viability and cell division, induced apoptosis, and arrested the cell cycle of T47D cells by triple inhibiting the p-Nrf2, p-ERα, and p-PI3K/AKT/mTOR signaling pathway. Molecular docking analysis confirmed that the possible mechanism of EL to reduce T47D cell growth was by inhibiting ERα and Nrf2-complex, resulting in the reduction in the crosstalk effect of Nrf2, ERα and PI3K/AKT/mTOR pathways. CONCLUSION: The combination of leaf extracts from E. scaber and P. macrocarpa caused cell death in breast cancer cells T47D and not in normal cells TIG-1; hence has the potential to show anticancer efficacy in preclinical and clinical trials.

Comparative Study of Antiproliferative Activity in Different Plant Parts of Phaleria macrocarpa and the Underlying Mechanism of Action.

Phaleria macrocarpa is a medicinal plant widely used in Indonesian folk medicine to treat several diseases, including cancer. However, the comparative evaluation of various plant parts of P. macrocarpa has not been studied for their anticancer properties on breast cancer. The study aimed to assess the antiproliferative activity of the ethanol extract of various parts of Phaleria macrocarpa against T47D human breast cancer cell lines. Several parts of P. macrocarpa, including pericarp, mesocarp, seed, and leaf, were used to determine the most potent plant part to inhibit the growth of T47D cells. The cytotoxic effects of each plant part were evaluated by WST-1 assay. The apoptotic level of T47D cells was determined by annexin V-FITC-PI and DNA fragmentation assay. Propidium iodide staining and the CFSE assay were used to examine the effect of each extract on cell cycle distribution and cell division, respectively. The relative number of caspase-3, Bax, and Bcl-2 was analyzed by flow cytometry technique. WST-1 assay revealed that P. macrocarpa leaves exhibited the most potent antiproliferative activity (p < 0.05) compared to other plant parts with selectivity only to T47D cells. P. macrocarpa leaves extract effectively induced apoptosis, inhibited proliferation, and arrested the cell cycle of T47D cells. The relative number of caspase-3 was significantly (p < 0.05) increased after being treated with P. macrocarpa leaf extract. P. macrocarpa leaf extract also leads to the dose-dependent accumulation in the Bax/Bcl-2 ratio due to upregulation of Bax and downregulation of Bcl-2. The overall results indicated that P. macrocarpa leaves could inhibit the proliferation of T47D cells and trigger apoptosis through caspase-3 activation and Bax/Bcl regulation. Therefore, P. macrocarpa leaves can be used for breast cancer therapy.

Metabarcoding dataset on the elicitation of Soybean and Mungbean using Ragi Tape as elicitors for enhancing secondary metabolites production.

Ragi Tape (RT) is commonly used as the microbial starter for various Indonesian traditional food. Consist of diverse microbial populations, RT has excellent agents as elicitors to augment the bioactive compounds in Soybean (SB) and Mungbean (MB). Metabarcoding analysis has shown to identify the microbial population involved in elicitation using RT. Inoculated RT from SB and MB were collected to extract the microbial DNA for the post-elicitation group. In comparison, DNA extraction from powdered RT was conducted as a pre-elicitation group. The total DNA were then sequenced by using the MiSeq Illumina platform with 16S rDNA gene region V3-V4 and 18S rDNA gene region V4 as a biomarker for bacterial and fungal identification, correspondingly. The obtained raw-data sequences were then analyzed using QIIME2 pipeline. According to the number of the acquired sequences, the 18S sequencing yielded more DNA strands than the 16S sequencing. However, the number of assigned OTUs was higher in 16S sequences than 18S sequences. From the perspective of the sample, RT has larger distinctive taxa, which were not identified in other samples. This metagenome data will provide fundamental information on RT employment in the elicitation process and further understanding of elicitation mechanisms using RT as biotic elicitors. The data is available at the BioProject database under the NCBI domain with accession no. PRJNA767401.

Lactobacillus plantarum FNCC 0137 fermented red Moringa oleifera exhibits protective effects in mice challenged with Salmonella typhi via TLR3/TLR4 inhibition and down-regulation of proinflammatory cytokines.

BACKGROUND: Salmonella typhi is a foodborne pathogenic bacterium that threatens health. S. typhi infection exacerbated the antibiotic resistance problem that needs alternative strategies. Moringa oleifera possesses anti-inflammatory and antimicrobial effects. However, there is a lack of information about the pharmacological value of red M. oleifera. The fermentation of red M. oleifera leaves extract (RMOL) is expected to add to its nutritional value. OBJECTIVE: The present study aimed to evaluate non-fermented RMOL (NRMOL) and fermented RMOL (FRMOL) effects on S. typhi infection in mice. MATERIALS AND METHODS: Female Balb/C mice were randomly divided into eight groups. The treatment groups were orally administered with NRMOL or FRMOL at doses 14, 42, and 84 mg/kg BW during the 28 days experimental period. Then S. typhi was introduced to mice through intraperitoneal injection except in the healthy groups. The NRMOL or FRMOL administration was continued for the next seven days. Cells that expressed CD11b+ TLR3+, CD11b+TLR4+, CD11b+IL-6+, CD11b+IL-17+, CD11b+TNF-a+, and CD4+CD25+CD62L+ were assessed by flow cytometry. RESULTS: Our result suggested that NRMOL and FRMOL extracts significantly reduced (p <0.05) the expression of CD11b+TLR3+, CD11b+TLR4+, CD11b+IL-6+, CD11b+IL-17+, and CD11b+TNF-α+ subsets. In contrast, NRMOL and FRMOL extracts significantly increased (p <0.05) the expression of CD4+CD25+CD62L+ subsets. NRMOL at dose 14 and 42 mg/kg BW was more effective compared to FRMOL in reducing the expression of CD11b+TLR3+, CD11b+TLR4+, and CD11b+TNF-α+ subsets. CONCLUSION: Our findings demonstrated that NRMOL and FRMOL extracts could be promising agents for protection against S. typhi infection via modulation of TLR3/TLR4, regulatory T cells, and proinflammatory cytokines.

Single-bulb garlic oil regulates toll-like receptors and Nrf2 cross-talk and IL-17 production in mice fed with high-fat diet.

The present study aimed to evaluate the effect of single-bulb garlic oil (SGO) on toll-like receptors 3 and 4 (TLR3 and TLR 4) and nuclear erythroid factor-like 2 (Nrf2) signaling pathway resulted from a high-fat diet and its underlying mechanism. Twenty-four Balb/c mice allocated into six groups: 1) N: mice fed with standard chow; 2) HFD: mice fed a high-fat diet for 45 days without any treatment; 3) HFD + Simv: mice fed a high-fat diet for 45 days and treated with simvastatin; 4-6) HFD + SGO 100, 200, 400 (mice fed a high-fat diet for 45 days and treated with single-bulb garlic oil at dose: 100, 200, and 400 mg/kg body weight for 30 days), respectively. At the end of treatment, spleen and hepar were isolated. The flow cytometry analysis was performed to analyze the relative number of nrf2, superoxide dismutase (SOD), malondialdehyde (MDA), TLR3, TLR4 and interleukin (IL-17). The results showed that HFD induction significantly reduced Nrf-2 and antioxidant enzyme levels. Furthermore, HFD induction increased TLR3 and TLR4 signaling and IL-17 production. Interestingly, 200 mg/kg BW of SGO increased the relative number Nrf-2 followed by SOD and HO-1 elevation at a dose of 100 mg/kg BW. SGO100 notably decrease the relative number of TLR3 (CD11b+TLR3+) and TLR4 (CD11b+TLR4+). The production of IL-17 by CD4 and CD8 were also reduced after receiving SGO at 200 mg/kg BW. This study suggests that the protective effect of SGO treatment on HFD mice was achieved by modulating TLR-Nrf2 cross-talks and decreasing IL-17 production. Our findings support a potential beneficial role of SGO for treating metabolic disease caused by a high-fat diet.

The combination of Elephantopus scaber and Sauropus androgynus promotes erythroid lineages and modulates follicle-stimulating hormone and luteinizing hormone levels in pregnant mice infected with Escherichia coli.

BACKGROUND AND AIM: Escherichia coli infection produces an adverse effect on the erythrocyte lineage and hormone levels during pregnancy. This study aimed to evaluate the effects of Elephantopus scaber (ES) and Sauropus androgynus (SA) in combination on circulating follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels and erythropoiesis changes in E. coli-infected pregnant mice. MATERIALS AND METHODS: Female Balb/c mice were mated with normal male mice and pregnancies were identified by the formation of vaginal plugs. Twenty-eight pregnant mice were divided randomly into seven groups: A control group (N), E. coli-infected pregnant mice (K+), and infected pregnant mice received the following five treatments: (1) Only ES; (2) ESSA1 (75:25); (3) ESSA2 (50:50); (4) ESSA3 (25:75); and (5) only SA, beginning from the 1st to the 16th day of pregnancy. Pregnant mice were infected with 107 CFU/mL of E. coli on day 4. Blood serum was collected on days 8, 12, and 16 of pregnancy and LH and FSH levels were measured by enzyme-linked immunosorbent assay. Bone marrow was isolated to determine the relative number of TER-119+VLA4+ and TER-119+CD34+ using flow cytometry. RESULTS: The ESSA1 and SA groups exhibited a marked increase in LH levels. The combination of ES and SA administered at a 25:75 ratio (ESSA3) altered FSH levels and the relative number of TER-119+VLA4+ in infected pregnant mice. Combined with SA at an equal ratio (50:50), ESSA2 group exhibited a significant increase in the expression of TER119+CD34+ compared with the other treatment groups. CONCLUSION: ES and SA combined at a ratio of 25:75 exhibited optimal results in altering hormonal and erythropoiesis in infected pregnant mice.

Suppression of hypoxia and inflammatory pathways by Phyllanthus niruri extract inhibits angiogenesis in DMBA-induced breast cancer mice.

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-1α (HIF-1α) 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-1α 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-1α), 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.

Elicited soybean extract attenuates proinflammatory cytokines expression by modulating TLR3/TLR4 activation in high-fat, high-fructose diet mice.

BACKGROUND: The high-fat, high-fructose diet (HFFD) provokes overnutrition and inflammation directly, mainly through Toll-like receptors (TLRs). Soybean (Glycine max L.) contains isoflavone that can be transformed into glyceollin by microbial and physical stimuli. Glyceollin possesses many beneficial effects on health. OBJECTIVE: This study evaluates the beneficial effect of soybean extract elicited by Saccharomyces cerevisiae and light (ESE) on dendritic cells (DCs) profile and naïve T cells in HFFD mice. MATERIALS AND METHODS: Female Balb/C mice were fed with HFFD for 24 weeks then orally administered with simvastatin 2.8 mg/kg BW or ESE 78, 104, and 130 mg/kg BW at the last four weeks. The expression of splenic CD11c+TLR3+, CD11c+TLR4+, NFκB+, CD11c+IL-17+, CD11c+TNF-α+, CD4+CD62L+, and CD8+CD62L+ subsets was measured by flow cytometry. The molecular docking has been measured using Pyrx 0.8, displayed in PyMol and Biovia Discovery Studio. RESULT: HFFD significantly increased CD11c+TLR3+, CD11c+TLR4+, NFκB+, CD11c+IL-17+, CD11c+TNF-α+ expression and decreased CD4+CD62L+ and CD8+CD62L+ (p < 0.05) compared to normal diet (ND) groups. ESE reduced CD11c+TLR3+, CD11c+TLR4+, thereby decreasing NFκB+, as well as decreased the CD11c+IL-17+, CD11c+TNF-α+, and restores CD4+CD62L+ and CD8+CD62L+ subsets in HFFD mice. Glyceollin II exhibited the best binding affinity with an average energy of -7.3 kcal/mol to TLR3 and -7.9 kcal/mol to TLR4. CONCLUSION: The bioactive compound in ESE act synergistically to modulate TLR3/TLR4 activation, reduced NFκB, IL-17, and TNF-α, and restores naïve T cells expression in HFFD mice. ESE was a favorable candidate to mitigate chronic inflammation.

Hormone-balancing and protective effect of combined extract of Sauropus androgynus and Elephantopus scaber against Escherichia coli-induced renal and hepatic necrosis in pregnant mice.

BACKGROUND: Elephantopus scaber (ES) and Sauropus androgynous (SA) have been frequently reported to possess antibacterial activity through in vitro, but in vivo studies about the protective effect of combined ES and SA have acquired less attention. OBJECTIVES: To evaluate protective effect of combined ethanol extract of ES and SA on hormone imbalance and renal and hepatic necrosis formation in Escherichia coli-infected pregnant mice. MATERIALS AND METHODS: A total of 28 pregnant Balb/c mice were divided into seven groups (n = 4): control, E. coli-infected pregnant mice, infected pregnant mice received 200 mg/kg ES, infected pregnant mice received combined 150 mg/kg ES and 37.5 mg/kg SA (75:25), 100 mg/kg ES and 75 mg/kg SA (50:50), 50 mg/kg ES and 112.5 mg/kg SA (25:75), and only 150 mg/kg SA. Pregnant mice were orally treated with combined ES and SA on day 1-4th of pregnancy. On the 4th day, mice were infected with 107 CFU/mL of E. coli and continuously treated with ES and SA until the 16th day of pregnancy. After treatment, the kidney and liver were prepared for histological examination using H&E staining. The blood serum was collected in each stage of pregnancy and measured by ELISA assays. RESULTS: Combined ES and SA gave an impact on altering the prolactin level. Combined ES and SA at ratio dose 75:25 was able to restore progesterone to normal levels (P < 0.05). The level of estradiol (E2) was relatively stable in the presence of E. coli and treatment. Treatment with 200 mg/kg ES, combined 50 mg/kg ES and 112.5 mg/kg SA (25:75) and 100 mg/kg ES and 75 mg/kg SA (50:50) demonstrated an immunomodulatory effect on the Gr1+ cell of E. coli treated-pregnant mice. E. coli infection significantly increased renal tubules and hepatic necrosis in pregnant mice compared to control (P < 0.05). Combined SA and ES at ratio dose 75:25 significantly demonstrated remarkable renal and hepatic protection activity in infected pregnant mice. CONCLUSION: The present study provided the establishment of combined ES and SA could be used to invent potent hormonal balancing agent and hepato-renal protective agent in infected pregnant mice.

Assessing the Immunomodulatory Activity of Ethanol Extract of Sambucus javanica Berries and Leaves in Chloramphenicol-Induced Aplastic Anemia Mouse Model.

Aplastic anemia, life-threatened disease, is a hematologic disorder characterised by bone marrow hypoplasia. Multiple modalities such as bone marrow transplantation and immunosuppression treatment have been proposed to ameliorate this entity, however it remains ineffective. Sambucus, a group of herb plants, possesses a broad spectrum of medicinal properties such as antioxidant, insulin-like activity, anticancer and antiviral. However, the study about its activity toward aplastic anemia incidence is based on limited data. Thus, the research aim of this study was to evaluate the immunomodulatory activities of Sambucus javanica in chloramphenicol-induced anemia aplastic mouse model. In this present study, BALB/c mice were administrated with chloramphenicol (CMP) to induce aplastic anemia then followed by S. javanica extracts treatment. Additionally, cellular and molecular aspects were evaluated by flow cytometry and Hematoxylin-Eosin staining. Further analysis showed that S. javanica extracts could promote the population number of regulatory T-cells and naive cytotoxic T-cells. Moreover, those extract also reduced the inflammation and necrotic incidence in CMP-induced mouse aplastic anemia model. Together, these results suggest that S. javanica has therapeutically effect to aplastic anemia by altering the immune system as an immunomodulatory agent.

Analysis of lymphocyte T(CD4+) cells expression on severe early childhood caries and free caries.

Early childhood caries (ECC) is still one of the many diseases found in children throughout the world. Cariogenic bacteria are a significant risk factor for ECC associated with early colonization and high levels of cariogenic microbes (Streptococcus mutans, S. mutans). Lymphocyte T (CD4+) cells known as helper T cells, are effector cells for mediated host immunity. Naive T cells (CD4+) must be activated to initiate effector function. This activation occurs through interaction with professional antigen- presenting cells (pro-APC), especially dendritic cells that lead to intracellular pathways that regulate T cell receptor (TCR) more specifically against antigen in T cells. Lymphocyte cells from samples were collected from severe early childhood caries (S-ECC) and Free caries aged 5 to 6 years. The subjects were instructed to gargle 10 mL of sterile NaCl 1.5% solution for 30 seconds, and expectorate it into a sterile glass then analyzing T lymphocyte cell (CD4+) expression using flow cytometry. Lymphocyte T (CD4+) cell expression at SECC (6.2525±64482) while in free caries (8.4138±1.10397) with P-value (P=0. 000). Conclusion of lymphocyte T (CD4+) cells expression at S-ECC is lower than that occurring in free caries.

Single garlic oil modulates T cells activation and proinflammatory cytokine in mice with high fat diet.

BACKGROUND: Hyperlipidemia triggers atherosclerosis by involving immune cells, such as T-cells. T-cells plays a role in worsening conditions during a high-fat diet (HFD). OBJECTIVE: The research aimed to analyze the role of single garlic oil (SGO) on T-cells activation and its proinflammatory cytokine expression in HFD mice. METHODS: Mice were divided into six groups: ND (normal diet); HFD (high-fat diet without treatment); HFD + Simv (HFD + simvastatin 2.6 mg/kg body weight); and HFD + SGO 1-3 (high-fat diet + single garlic oil in a dose of 12.5, 25, and 50 mg/kg body weight), respectively. Treatments were orally given every day for 45 days. At the end of treatments, lymphocytes were isolated from mice spleen. The relative number of T-cells and proinflammatory cytokines were measured using flow-cytometry and analyzed using one-way ANOVA (p < 0.05). RESULT: Our result indicated that HFD mice had lower naive T cells (CD4+CD62L+) than normal mice (p < 0.05). SGO treatment in HFD mice increased the relative number of naïve T cells. HFD treatment increased the expression of TNF-α and IFN-γ through NF-κB expression. Furthermore, SGO treatment improved the expression of TNF-α and IFN-γ. CONCLUSION: Our study suggests that SGO could act as a promising prospect for therapy to improve chronic inflammation in a HFD.

Detection of class 1 integron-associated gene cassettes and tetracycline resistance genes in Escherichia coli isolated from ready to eat vegetables.

BACKGROUND: Ready to eat (RTE) vegetables are easily accessible healthy foods that are commonly consumed globally, including in Indonesia. However, these RTE vegetables contain potential contamination from pathogens and multi-drug resistant bacteria. Therefore, in the present study, we examined the presence of tetracycline-resistant E. coli (TRE) isolates from RTE vegetables. METHODS: Susceptibility to antimicrobial agents was determined using the Kirby-Bauer disc diffusion method. Characterisation of antibiotic resistant genes was performed using PCR and sequencing of tetracycline resistant gene, integron and gene cassette from the TRE isolates. RESULTS: The isolates collected in this study were resistant not only to tetracycline, but also to streptomycin. Some isolates also displayed resistance to kanamycin (77.8%), chloramphenicol (11.1%), and ciprofloxacin (5.6%). All of the isolates contained integrons (intI1) and the tetA gene; tetB was not detected in our study. Further analysis showed that some isolates (38.8%) contained the dfrA7 gene cassette, which encodes dihydrofolate reductase, which is responsible for resistance to trimethoprim. Of all the isolates that presented integrons, 11 isolates (61.1%) did not carry gene cassettes. These empty integrons have the potential to convert themselves rapidly into multigraviton strains. CONCLUSIONS: TRE isolates contain the tetA gene and integron 1. Only 38.8% of the isolates that have been identified contain the dfrA7 gene cassette, which is responsible for trimethoprim antibiotic resistance. Further identification of genes conferring resistance to other antibiotics is necessary to better characterise antibiotic resistance.

The significance of Agaricus blazei as an immunomodulator of the level of IL-17 in Balb/C mice with atherosclerosis.

Atherosclerosis is a disease caused by an inflammatory response which involved the interaction between endothelial cells, macrophages and lymphocytes, and is closely related to IL-17 regulation. This study is important to investigate the activity of Agaricusblazei in modulating the immunological activity based on the profile of CD4+IL-17+, CD8+IL-17+, and CD11b+ IL-17+ in high-fat diet (HFD)-induced Balb/c mice. Mice in dietary groups were fed with HFD and then fed with A. blazei extract with three different doses including D1 (100 mg/kg BW), D2 (200 mg/kg BW), and D3 (400 mg/kg BW) once a day for 12 weeks. The cells were analyzed using flow cytometry and tested statistically with one-way ANOVA with α = 0.05 by using SPSS 16.00 software. The results showed that mice with HFD treatment had a higher level of Lp-PLA2 (atherosclerosis marker) compared with the control group (data not shown). The level of IL-17 in the atherosclerotic mice in the D1 group was significantly depleted compared to the control group. Of the three doses above, D1 may be an optimal dose to minimize or prevent the damage from atherosclerosis than the other doses.

The isolation of exopolysaccharide-producing lactic acid bacteria from lontar (Borassus flabellifer L.) sap.

BACKGROUND AND OBJECTIVES: Lontar (Borassus flabellifer L.) is widely grown in Indonesia and one of its products is palm sap. Palm sap contains a high level of sugar, making it suitable as a medium to increase the lactic acid bacteria (LAB) production of exopolysaccharides (EPS). This study aimed to isolate the EPS-producing LAB from palm sap and evaluate its EPS production. LAB isolation was carried out on MRS agar containing 0.5% CaCO3. MATERIALS AND METHODS: The screening and production of EPS were carried out on MRS media supplemented with 10% sucrose. The molecular identification of the selected EPS-producing LAB was based on 16S rDNA. A quantitative analysis of EPS polymer dry mass and total sugar was conducted using one-way ANOVA. RESULTS: In this study, five EPS-producing LABs were found: Fructobacillus fructosus N4, Leuconostoc mesenteroides N5, Leuconostoc mesenteroides N7, Leuconostoc mesenteroides N9, and Fructobacillus fructosus N10. The highest EPS yield in liquid media was 10.997 ± 1.591 g/L by Leuconostoc mesenteroides N7, whereas the lowest was 4.505 ± 0.459 g/L by Fructobacillus fructosus N10. CONCLUSION: This study found Fructobacillus fructosus strains as EPS producers that have never been reported before.

Immunomodulatory Activities of Sambucus javanica Extracts in DMBA-Exposed BALB/c Mouse.

Purpose: Accumulating evidence shows the genus of Sambucus exerts a broad spectrum of medicinal potencies such as anticancer, antiviral, antibacterial, and antidiabetes. Based on the previous studies, we hypothesized that bioactive compounds of Sambucus might alter several biological systems, including the immune system. Therefore, this study extensively aimed to evaluate the immunomodulatory activities of Sambucus javanica extracts in 7,12-dimethylbenz[a] anthracene (DMBA)-treated BALB/c mouse. Methods: The experimental mice were orally administrated with 2.8 mg.kg-1 BW of DMBA for ten times within a month. After that, the mice were treated by S. javanica berries and leaves extracts for 2 weeks. Subsequently, the inflammation rate was evaluated by using flow cytometry analysis, whereas the necrosis incidences were observed by hematoxylin & eosin staining. Results: Based on the results, we found the expression of tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-É£) were increased however after treated by S. javanica berries and leaves extracts were significantly decreased. In the same way, necrosis incidence was increased in the DMBA-treated group however it was diminished with S. javanica extracts treatment. Conclusion: Together, these results suggested that S. javanica extracts have immunomodulatory activities to suppress inflammation and reduce necrosis incidence in experimental mice.

Anti-inflammatory activity of elicited soybean (Glycine max) extract on Balb/C mice (Mus musculus) with high-fat and -fructose diet.

Obesity causes adipocyte hypertrophy, which leads to cell death. Consequently, macrophages and lymphocytes infiltrate into the adipose tissue and elevate pro-inflammatory cytokine production through TLR activation. This study aimed to determine the efficacy of soybean extract, which was elicited by Saccharomyces cerevisiae and light, as an anti-inflammatory agent in mice with a high-fat and -fructose diet (HFFD). The elicited soybean extract (ESE) was administered orally to mice for four weeks after being given an HFFD for 20 weeks. Three different doses were used: (1) low-dose (78 mg/kg BW); (2) normal dose (104 mg/kg BW); and (3) high dose (130 mg/kg BW). HFFD mice model treated with simvastatin 2.8 mg/kg BW considered as drug control. After 24 weeks, the lymphocytes were isolated and the relative number of CD4+TLR3+ T, CD4+TLR4+ T, CD4+TNF-α + T, and CD4+IFN-γ + T cells were analysed using flow cytometry. The results showed that the HFFD mouse model had an increased number of CD4+TLR3+ T, CD4+TLR4+ T, CD4+TNF-α + T, and CD4+IFN-γ + T cells. ESE administration decreased the relative number of CD4+TLR3+ T, CD4+TLR4+ T, CD4+TNF-α + T, and CD4+IFN-γ + T cells. The normal dose of ESE is the most effective dose in suppressing inflammation compared to positive controls. ESE 104 mg/kg BW can be considered as an alternative herbal medicine that may suppress inflammation in HFFD mice.