Using metabolomics to discover the immunomodulator activity of food plants.

Many edible plants exhibit immunomodulator activities that have beneficial effects on human health. These activities include the ability to activate, multiply, or suppress elements of the immune response. Some of these plants promote health by strengthening host defences against different diseases. In this article, we provide a comprehensive review of the constituents of several edible plants, their immunomodulatory activity, and mechanism of actions for Carica papaya, Coffea sp, Asparagus cochinchinensis, Dioscorea alata, beans, mushrooms, herbs, spices, and several vegetables. The studies reported here are pre-clinical (in vitro and in vivo) and clinical studies (limited in number). The bioactive compounds responsible for the immunomodulator activity of these plants were yet to be identified. This is because the plant is naturally a complex mixture, whilst the immune system is also an intricate system involving many cells and cytokines/chemokines. Metabolomics is a key tool for conducting global profiling of metabolites in a complex system. Therefore, it offers the ability to identify the presence of compounds in plant extracts associated with their immunomodulation effects. Likewise, metabolomics can also be used to detect any changes to metabolites in the cell as a response to treatment. Therefore, affected metabolic pathways that lead to the activation of certain immune responses can be determined from one single experiment. However, we found in this review that the use of a metabolomics approach is not yet fully developed for an immunomodulator study of food plants. This is important for the direction of future research in this field because unlike medicinal plants, food plants are consumed on a regular basis in small amounts with more obvious effects on the immune system. Information about possible bioactive compounds, their interactions (synergism, antagonism), and how the human body responds to them should be studied in a more holistic way.

Angiotensin-I-Converting Enzyme Inhibitory Peptides in Goat Milk Fermented by Lactic Acid Bacteria Isolated from Fermented Food and Breast Milk.

In this study, angiotensin-I-converting enzyme inhibitory (ACEI) activity was evaluated in fermented goat milk fermented by lactic acid bacteria (LAB) from fermented foods and breast milk. Furthermore, the potential for ACEI peptides was identified in fermented goat milk with the highest ACEI activity. The proteolytic specificity of LAB was also evaluated. The 2% isolate was inoculated into reconstituted goat milk (11%, w/v), then incubated at 37°C until pH 4.6 was reached. The supernatant produced by centrifugation was analyzed for ACEI activity and total peptide. Viable cell counts of LAB and titratable acidity were also evaluated after fermentation. Peptide identification was carried out using nano liquid chromatography mass spectrometry (LC-MS/MS), and potential as an ACEI peptide was carried out based on a literature review. The result revealed that ACEI activity was produced in all samples (20.44%-60.33%). Fermented goat milk of Lc. lactis ssp. lactis BD17 produced the highest ACEI activity (60.33%; IC50 0.297±0.10 mg/mL) after 48 h incubation, viable cell counts >8 Log CFU/mL, and peptide content of 4.037±0.27/mL. A total of 261 peptides were released, predominantly derived from casein (93%). The proteolytic specificity of Lc. lactis ssp. lactis BD17 through cleavage on the amino acid tyrosine, leucine, glutamic acid, and proline. A total of 21 peptides were identified as ACEI peptides. This study showed that one of the isolates from fermented food, namely Lc. lactis ssp. lactis BD17, has the potential as a starter culture for the production of fermented goat milk which has functional properties as a source of antihypertensive peptides.

Immunomodulatory and Antioxidant Activities of Select Indonesian Vegetables, Herbs, and Spices on Human Lymphocytes.

Edible plants have attracted increasing attention as functional foods as they are rich in bioactive compounds with health benefits, including antioxidant and immunomodulatory activities. However, scientific evidence of these health effects is limited. This study is aimed at determining antioxidant and immunomodulatory activities of 25 select vegetables, herbs, and spices commonly consumed in Indonesia. Phytochemical profiles were determined by measuring total flavonoid content and 1H-NMR. Human blood lymphocyte cells were used to probe the immunomodulatory potency and treated with the methanol extract of these vegetables, herbs, and spices. The results showed the enhanced propensity for all tested plant extracts to stimulate lymphocyte proliferation, except Pandanus amaryllifolius. Etlingera elatior, Ocimum xcitriodorum, Kaempferia galanga, and Apium graveolens had the highest lymphocyte cell proliferation stimulation index (SI) at concentrations of 41.67, 16.67, 4.17, and 2.5 mg/mL culture, respectively (SI 2.21 ± 0.05, 2.62 ± 0.12, 3 ± 0.05, and 2.64 ± 0.07, respectively). The NMR spectra of these four most potent plants showed low peaks in the aromatic/phenolic area and several other peaks indicating the presence of terpenoid, steroid, amino acid, and sugar compounds. The results demonstrate the immunomodulatory potential of all vegetables, herbs, and spices, except P. amaryllifolius, although this potential did not necessarily correlate with flavonoid content and antioxidant activity. Nevertheless, this research showed promising health effect, particularly immunomodulation, of the various local plants. Further elaboration on the specific immunomodulatory activity will be interesting.

Angiotensin-I-converting enzyme inhibitory peptides in milk fermented by indigenous lactic acid bacteria.

BACKGROUND AND AIM: Fermented milk can be used to produce antihypertensive peptides. Lactic acid bacteria (LAB) with its proteolytic system hydrolyze milk protein during fermentation to produce several peptides, which include antihypertensive bioactive peptides. This study aimed to investigate the ability of indigenous LAB for the production of angiotensin-I-converting enzyme inhibitory (ACE-I) peptides in fermented milk and to characterize the ACEI peptides. MATERIALS AND METHODS: Reconstituted milk (11%) inoculated with ten LAB isolates, and then incubated at 37°C until it reaches pH 4.6. The evaluation was carried out for LAB count, lactic acid concentration, peptide content, and ACE-I activity. The low molecular weight (MW) peptides (<3 kDa) were identified using Nano LC Ultimate 3000 series system Tandem Q Exactive Plus Orbitrap high-resolution mass spectrometry. RESULTS: The result showed that the ten LAB isolates were able to produce ACE-I in fermented milk with the activities in the range of 22.78±2.55-57.36±5.40%. The activity of ACE-I above 50% produced by Lactobacillus delbrueckii BD7, Lactococcus lactis ssp. lactis BD17, and Lactobacillus kefiri YK4 and JK17, with the highest activity of ACE-I produced by L. kefiri YK4 (IC50 0.261 mg/mL) and L. kefiri JK17 (IC50 0.308 mg/mL). Results of peptide identification showed that L. kefiri YK 4 could release as many as 1329, while L. kefiri JK 17 could release 174 peptides. The peptides produced were 95% derived from casein. The other peptides were from ά-lactalbumin, ß-lactoglobulin, and serum amyloid A. The peptides produced consisted of 6-19 amino acid residues, with MWs of 634-2079 Dalton and detected at 317-1093 m/z. A total of 30 peptides have been recognized based on literature searches as ACE-I peptides (sequence similarity: 100%). CONCLUSION: L. kefiri YK4 and JK17 are the potential to be used as starter cultures to produce the bioactive peptide as ACE-I in fermented milk.

The proportion-ratio on dipeptidyl aminopeptidase-4 (DP-4) inhibition by gelatin compared to synthetic sitagliptin.

The current study aims to determine the inhibition activity gelatin against dipeptidyl aminopeptidase 4 (DP-4). Two commercial gelatins, i.e., bovine and fish skin gelatin and one extracted (in our laboratory) gelatin, i.e., fish bone gelatin were selected for analysis. Each gelatin have same protein pattern (75-245 kDa) on sodium dodecyl sulfate polyacrylamide gel electrophoresis with mean of protein concentration of 1.72 mg/mL. The inhibition activity was measured on the capacity to inhibit DP-4 by using Gly-Pro-p-nitroanilide as their substrate. The sitagliptin was used as standard comparison. Based on the percent inhibition, gelatin has been shown to be the prospective DP-4 inhibitor.

Dietary exposure to sulfites in Indonesians.

BACKGROUND AND OBJECTIVES: Information on dietary exposure to sulfites as preservative in consumer is needed as a scientific base for food safety policy making. The objective of this research was to conduct dietary exposure assessment to sulfites in consumers by using a deterministic method. METHODS AND STUDY DESIGN: The scope of work was identification of food products containing sulfites, determination of food consumption data from the individual food consumption survey report of 2014, determination of sulfite concentration in food, and calculation of sulfite exposure. RESULTS: 3,428 (9%) of 37,613 food products registered in National Agency of Drug and Food Control (2012-2015) may contain sulfite. The most used sulfite in food products was sodium metabisulfite. The mean of food containing sulfite consumption in all age groups was 131.4 g/person/day. The estimation of total exposure for all age groups were 0.27 mg/kgBW/day (38.6% ADI), 0.25 mg/kgBW/day (35.7% ADI) and 0.08 mg/kgBW/day (11.4% ADI) by using concentrations of Maximum Permitted Limit, reported maximum used level and reported maximum product test result, respectively. Food category contributed to the highest exposure in all age groups was spices, condiments, vinegar, powder or mixture for soups and broths, and other soy sauce category. CONCLUSIONS: The highest total exposure to sulfites was found in 0-59 month age group. The highest total exposure for the MPL (0.79 mg/kgBW/day) and the reported maximum used level (0.73 mg/kgBW/day) exceeded 112.9% ADI and 104.3% ADI, respectively while the exposure using reported maximum test result was still below ADI (0.25 mg/kgBW/day or 35.7% ADI).

Plant Sterol Esters in Extruded Food Model Inhibits Colon Carcinogenesis by Suppressing Inflammation and Stimulating Apoptosis.

Plant sterols in their free forms are known to inhibit colon cancer. Whether these activities persist when compounds are incorporated into processed food is not reported yet. This study aimed to test the ability of plant sterol esters (PSE) incorporated into a nonpuffed extruded food (NPE) model to inhibit colon carcinogenesis. PSE was added into NPE at four concentrations (0.0%, 0.7%, 1.4%, and 2.1%). PSE-NPE activity was tested in azoxymethane/dextran sodium sulfate-induced Balb/c mice. The groups given PSE-NPE did not show any colon tumor formation. Immunohistochemistry results revealed that the group fed with 1.4% PSE had the lowest histoscore for cyclooxygenase-2 expression and the highest histoscore for cleaved caspase-3, cleaved caspase-8, and cleaved caspase-9expressions. The results of this study indicated that even after incorporation into a food system, which is processed using high pressure and temperature, PSE retained its chemopreventive activity. The proposed mechanisms are by suppressing inflammation and inducing apoptosis.