Anti-inflammatory effect of the immature peel extract of <Emphasis Type="Italic">Jinkyool</Emphasis> (<Emphasis Type="Italic">Citrus sunki</Emphasis> Hort. ex Tanaka)

The peel of jinkyool (Citrus sunki Hort. ex Tanaka) has been widely used in traditional Asian medicine for treatment of a number of diseases, including indigestion and bronchial asthma. In the present study, we compared the flavonoids content and anti-inflammatory activities of ethanolic peel extracts from both immature and mature fruits. Comparing to the mature peel extract (MPE), the immature peel extract (IPE) contained more abundant flavonoids. IPE more effectively suppressed intracellular reactive oxygen species (ROS) generation (IPE, 67.6±1.2%; MPE, 78.9±2.4% at 300 μg/mL of control), nitric oxide (NO) production, and inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expression, as well as interleukin (IL)-1β, IL-6, and tumor necrosis factor-α (TNF-α) mRNA expression in lipopolysaccharide-stimulated RAW 264.7 cells. Additionally, IPE reduced c-Jun NH₂-terminal kinase (JNK) and p38 mitogen-activated kinase (p38) activation, suggesting that IPE may exert an anti-inflammatory effect by suppressing pro-inflammatory cytokines that suppress activation of mitogen-activated protein kinases (MAPKs). Taken together, these results indicate that IPE has potential for use as an anti-inflammatory agent.     

Estrogenic effects of various extracts from <Emphasis Type="Italic">Chamdanggui</Emphasis> (<Emphasis Type="Italic">Angelica gigas</Emphasis> Nakai) and <Emphasis Type="Italic">sogdan</Emphasis> (<Emphasis Type="Italic">Phlomis umbrosa</Emphasis> Turcz)

The various extracts from chamdanggui (Angelica gigas Nakai) and sogdan (Phlomis umbrosa Turcz) were evaluated for estrogenic activity and characterized according to HPLC profile. Chamdanggui and sogdan were individually extracted with 4 solvents (hot water, 70% ethanol, n-butanol, and dichloromethane) of differing polarities. Estrogenic activity was determined by E-screen using an estrogen-dependent MCF-7 BUS cell. Although almost all extracts showed estrogenic effects in a concentrationdependent manner, the hot water extract from chamdanggui (250 μg/mL) had the higher effect (138%). Among 90 fractions using HPLC separation of the hot water extract from chamdanggui, fraction 21 and 28 produced the highest estrogenic effects of 178 and 163% at 10 μg/mL, respectively. The results imply that the hot water extract from chamdanggui could be useful as an alternative hormone replacement therapy.     

Ethanolic fermentation from red ginseng extract using <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> and <Emphasis Type="Italic">Saccharomyces carlsbergensis</Emphasis>

The pH of red ginseng extracts fermented with Saccharomyces cerevisiae and Saccharomyces carlsbergensis decreased rapidly during 3 days of fermentation, with no further significant change thereafter. After 20 days of fermentation, a relatively small difference remained in the acidity of extracts fermented with S. cerevisiae (0.54%) and S. carlsbergensis (0.58%). Reducing sugar in the S. cerevisiae and S. carlsbergensis extracts decreased from 258.6 to 45.4 and 43.2 mg/mL glucose equivalents, respectively; and ethanol contents increased from 1.5% at day 0 to 16.0 and 15.0%, respectively, at day 20. Ginsenosides Rb₁, Rb₂, Rc, Re, Rf, and Rg₁ decreased during the fermentation with S. cerevisiae, but Rd and Rg₃ increased by day 12. Ginsenosides Rb₁, Rb₂, Rc, Re, and Rg₁ decreased gradually in the extract with S. carlsbergensis, but Rd and Rg₃ were increased at day 6 and 9, respectively.     

Tyramine production of technological important strains of <Emphasis Type="Italic">Lactobacillus</Emphasis>, <Emphasis Type="Italic">Lactococcus</Emphasis> and <Emphasis Type="Italic">Streptococcus</Emphasis>

The aim of this paper was to study the biogenic amines (histamine, tyramine, putrescine, cadaverine, agmatine, spermine and spermidine) production of selected technological important lactic acid bacteria (strains of the genera Lactococcus, Lactobacillus and Streptococcus). Three methods (ion-exchange chromatography (IEC), PCR and cultivation method with pH indicator) were used. Within the 39 strains of lactic acid bacteria tested, the production of tyramine (formed by tyrosine decarboxylase) was detected in eight strains (3 strains of Lactococcus lactis subsp. lactis, three strains of Lactococcus lactis subsp. cremoris, 1 strain of Streptococcus thermophilus and 1 strain of Lactobacillus delbrueckii subsp. bulgaricus). The other tested biogenic amines were not detected. Cultivation in decarboxylation broth seems to be the least accurate method for the detection of biogenic amines due to enhanced risk of false-positive reactions. Therefore, in order to detect bacteria producing biogenic amines, the combination of PCR and chromatographic methods (e.g. IEC) can be recommended.     

Bioactive compounds and antioxidant activities of sprout soybean fermented with <Emphasis Type="Italic">Irpex lacteus</Emphasis> mycelia

To enhance the biological activities of sprout soybean, beans were treated with steaming (SS), germinating (GS), or roasting (RS) prior to fermentation with Irpex lacteus mycelia for 20 days. The total phenolic, flavonoid, isoflavone, and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity of each fermented bean preparation were examined every 5 days for 20 days. The total phenolic content of SS, GS, and RS preparations was 9.61, 10.23, and 10.46 mg/g, respectively, after 15 days of fermentation. These concentrations were approximately 4–5 folds higher compared to initial levels. The total flavonoid content was 8–9 folds higher than initial levels. The isoflavone content was highest in the RS sample (6.84 mg/g). The DPPH radical scavenging activity of beans fermented with I. lacteus mycelia was increased 2–8 folds after 20 days of fermentation. These results indicate that antioxidant activity components were increased by fermentation of I. lacteus mycelia irrespective of soybean treatments.     

In vivo study of antiallergenicity of ethanol extracts from <Emphasis Type="Italic">Sargassum tenerrimum</Emphasis>, <Emphasis Type="Italic">Sargassum cervicorne</Emphasis> and <Emphasis Type="Italic">Sargassum graminifolium turn</Emphasis>

Food allergy has becoming the serious threat in the world for which the search of an effective anti-allergic drug is the demand of time. Keeping in view of the potentiality of seaweeds, the ethanol extracts from Sargassum tenerrimum (ST), Sargassum cervicorne (SC), and Sargassum graminifolium turn (SG) have been studied in vivo for its antiallergenicity through passive cutaneous anaphylaxis (PCA) and active cutaneous anaphylaxis (ACA) in female BALB/c mice. Intraperitoneal administration of these ethanol extracts inhibit mouse PCA and ACA in a dose-dependent manner using ovalbumin (OVA) and shrimp allergen as triggering agents to induce allergenicity over mice. The extract of ST containing phlorotannin has been found most active over the suppression of PCA triggered by OVA and shrimp with IC₅₀ values of 25.64 and 40.98 mg/kg, respectively and an efficacy comparable to that of an anti-allergic drug disodiumcromoglycate. Similarly, ST inhibits ACA triggered by ova and shrimp allergen in the mouse, with 50% suppression at 25.5 and 43.53 mg/kg, respectively. The results presented here show that these extracts are active on the studied models among which ethanol extract of ST was the most potent, leading toward the promising development of a new class of anti-allergic drugs.     

Novel anticoagulant compound from fermented red alga <Emphasis Type="Italic">Pachymeniopsis elliptica</Emphasis>

Natural fermentation was tested as a method of releasing active compounds during screening for potential anticoagulant activity in three types of algae (Pachymeniopsis elliptica, Sargassum horneri, and Ulva pertusa). Freeze dried algae samples (2.5 g) were fermented by adding 75 g of sugar and 500 mL of water and thereafter kept at room temperature (25 °C) for 3 months. Activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT) were measured every 2 weeks for 3 months to determine the optimum time for the highest activity. Fermented P. elliptica, (which had the highest activity) was subjected to anion exchange chromatography (DEAE-cellulose) and sepharose 4B gel permeation chromatography. The purified sample was analyzed by agarose-gel electrophoresis and polyacrylamide gel electrophoresis (PAGE) to confirm the purification and to determine the molecular mass, respectively. The 360 μg/mL of purified compound (Mwt > 500,000 Da) had both APTT and PT activities (>1,000 s). However, at the concentrations of 180 μg/mL, purified compound and heparin showed 540 and >1,000 s APTT activity, respectively. Though, the purified compound of P. elliptica considered as a weaker anticoagulant than heparin, this purified anticoagulant polysaccharide could be considered as a good alternative source as an anticoagulant. Moreover, the technique of fermentation is an inexpensive and feasible, this purified anticoagulant polysaccharide compound could be used in pharmaceutical and biomedical industry. Further investigations need to be performed to determine the mechanism of this novel anticoagulant compound. The authors Prashani Mudika Ekanayake and Chamilani Nikapitiya contributed equally to this work.     

Molecular diversity among natural populations of <Emphasis Type="Italic">Lactobacillus paracasei</Emphasis> and <Emphasis Type="Italic">Lactobacillus plantarum</Emphasis>/<Emphasis Type="Italic">paraplantarum</Emphasis> strains isolated from autochthonous dairy products

The diversity of 87 Lactobacillus paracasei and Lactobacillus plantarum/paraplantarum strains, previously identified from different autochthonous dairy products, was investigated by phenotypic and genotypic approaches. The increased resolution obtained using phenotypic and genotypic characterization allowed the level of strain heterogeneity detection to be widened. Phenotypic diversity was evaluated by studying biochemical characteristics of technological interest, including antimicrobial and proteinase activities, resistance to nisin, aggregation ability, production of exopolysaccharides, acetoin and diacetyl, citrate utilization, and antibiotic susceptibility. Genotypic diversity was generally evaluated by PCR amplification of repetitive bacterial DNA element fingerprinting using the (GTG)₅ primer [(GTG)₅-PCR]. Moreover, in cases where strains were not discriminated by (GTG)₅-PCR combined with phenotypic analysis, pulsed-field gel electrophoresis (PFGE) analysis was performed. The results indicate that L. plantarum/paraplantarum and L. paracasei natural isolates from artisanal dairy products are a gold mine in terms of diversity of strains and could be potentially interesting to dairy companies for the formulation of functional starter cultures in the production of innovative foods.     

A New Multiplexed Real-Time PCR Assay to Detect <Emphasis Type="Italic">Campylobacter jejuni</Emphasis>, <Emphasis Type="Italic">C. coli</Emphasis>, <Emphasis Type="Italic">C. lari</Emphasis>, and <Emphasis Type="Italic">C. upsaliensis</Emphasis>

A new rapid method based on real-time PCR was developed to detect four thermophilic Campylobacter species (Campylobacter jejuni, Campylobacter coli, Campylobacter lari, and Campylobacter upsaliensis) in food samples. The assay targeted the bipA gene for C. upsaliensis and C. lari, whereas the gene encoding the ATP-binding protein CJE0832 was used to detect C. coli and C. jejuni. These genes were chosen for this assay due to their low variability and mutation rate at a species level. The multiplex PCR showed 100% inclusivity for all 25 thermophilic Campylobacter strains tested and 100% exclusivity for 38 non-targeted strains belonging to closely related species. The newly developed real-time PCR could detect down to 10² genomes/reaction and displayed efficiency above 97% for all species except for C. upsaliensis (90.1%). The method proved to be a reliable tool for food analysis, showing 100% sensitivity, 96% efficiency, and 92.45% specificity when validated against the gold standard method UNE-EN ISO 10272:2006 using 200 diverse food samples (meat, fish, fruits and vegetables, and raw milk). In artificially spiked samples, the detection limit of the method was 10 cfu/g in salad, 5 cfu/g in turkey meat, and 1 cfu/g in the rest of meat samples tested. Consequently, the newly designed molecular tool represents a quick and safe alternative to obtain reliable results concerning the presence/absence of the main thermophilic Campylobacter in any food sample.     

Thermal inactivation kinetics of quality-related enzymes in cauliflower (<Emphasis Type="Italic">Brassica oleracea</Emphasis> var. <Emphasis Type="Italic">botrytis</Emphasis><Emphasis Type="Bold">)</Emphasis>

Thermal inactivation of quality-related enzymes in both cauliflower crude enzyme extracts and fresh tissue samples was studied in temperature range 50–100 °C. For crude enzyme extracts, several parameters, reaction rate constants (k) and activation energy (E _a) as well as decimal reduction time (D) and (z) values, were used to characterize the thermal stability. The rates of inactivation were found to follow first-order inactivation kinetics. Activation energies varied between 101.18 and 208.42 kJ mol⁻¹ with z values of 10.59–24.09 °C. The examined kinetics indicated that lipoxygenase was the most heat resistant followed by peroxidase, polyphenol oxidase, pectin methyl esterase and ascorbic acid oxidase. Furthermore, the obtained results from the blanched fresh tissues indicated that inactivation of lipoxygenase secured disappearing of any other enzyme activities. Therefore, this study recommends using lipoxygenase as an indicator enzyme to optimize the thermal treatments of cauliflower products.     

<Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> fermented yam (<Emphasis Type="Italic">Dioscorea opposita</Emphasis> Thunb.) and its preventive effects on gastric lesion

This study was carried out to find the possibility of fermenting yam (Dioscorea opposita Thunb.) with Lactobacillus acidophilus and the protective role on stomach lesion of rats. The solutions of raw and extracted yam used were 2 and 6%, and they were fermented by L. acidophilus at 37°C for 0, 12, 24, and 36 h. At intervals (0, 12, 24, and 36 h), viscosity, titratable acidity (TA), allantoin, and diosgenin were examined. In the results, it was observed the most desirable data at 12 h with 6% fermented raw yam (FRY) without any change in the content of allantoin and diosgenin. In the animal study by rats (Sprague-Dawley), the inhibition ratios on gastric lesion were 53.41% in FRY. Based on these data, it confirmed that raw yam powder fermented by L. acidophilus would be a functional food with the supplement of viable cells and nutraceutical component of allantoin and diosgenin.     

Characterization of brewing microorganisms isolated from Korean traditional <Emphasis Type="Italic">nuruk</Emphasis> for <Emphasis Type="Italic">Cheongju</Emphasis> production

The objective of this study was to select and identify yeasts and molds isolated from traditional nuruk and to investigate their brewing characteristics for Cheongju production. The yeast strains Y190 (Accession ID-KACC 93251P), Y263 (Accession ID-KACC 93252P), and Y270 (Accession ID-KACC 93253P) showing high alcohol and flavor productivity were isolated and identified by phylogenetic inference based on an internal transcribed spacer 2 region sequence analysis. In addition, Aspergillus oryzae 83-10 (Accession ID-KACC 93254P) showing the highest enzyme activity was isolated. This study provides basic data for the production of Korean Cheongju and assesses the applicability of these three yeast strains and A. oryzae 83-10 isolated from traditional nuruk.     

Pressurized acidified water extraction of black carrot [<Emphasis Type="Italic">Daucus carota</Emphasis> ssp. <Emphasis Type="Italic">sativus</Emphasis> var. <Emphasis Type="Italic">atrorubens</Emphasis> Alef.] anthocyanins

The anthocyanins present in black carrot were extracted with pressurized water acidified with sulfuric, citric and lactic acids. Anthocyanin degradation became significant above 100 °C and there was no improvement when extraction pressure was increased to 100 bar. Therefore, the extraction from black carrot was carried out at temperatures 50, 75 and 100 °C under 50 bar pressure. The extraction efficiencies in terms of acylated and non-acylated anthocyanins were comparable for all three acids used to acidify water at 50 °C, while similar results were observed at 75 °C for both citric and lactic acids. Water acidified with lactic acid showed significantly higher extraction efficiency at 100 °C compared to water acidified with sulfuric and citric acids. Highest degree of polymerization together with increasing degree of browning was observed within extracts when sulfuric acid was used. On the other hand, when organic acids were used to acidify water, a higher extraction efficiency of anthocyanins, accompanied with a relatively low polymerization and browning was observed, with lactic acid giving the best results.     

Optimization for encapsulation of green tea (<Emphasis Type="Italic">Camellia</Emphasis><Emphasis Type="Italic">sinensis</Emphasis> L.) extract by spray drying technology

Green tea has been credited with providing a wide variety of health benefits like higher total antioxidant, cancer prevention, and anti-irritant etc. Microencapsulation technology is used to prevent antioxidant loss during processing which involves entrapment of active material into carrier material. The experiments were designed using response surface methodology based on three level two factor (green tea extract, 10–25 % and drying inlet air temperature, 120, 130 and 140°C) central composite design. The feed emulsion was prepared with green tea extract as core material and 40 % maltodextrin concentration as wall material with the ratio of core to wall (1:2) and it was fed into the spray dryer at varied drying inlet air temperature to get encapsulated green tea extract powder. The quality characteristics were analysed such as moisture content, total antioxidant activity and total phenolic content and the process conditions were optimized using desirability function methodology. At optimum spray drying condition moisture content, total antioxidant activity and total phenolic content were found to be 3.1 % w.b., 72.91 % free radical scavenging activity by DPPH method, 57.81 mg/g of dry matter, expressed as gallic acid equivalent respectively with desirability value of 0.92.     

Prevalence,genetic diversity,and antibiotic susceptibility of <Emphasis Type="Italic">Cronobacter</Emphasis> spp. (<Emphasis Type="Italic">Enterobacter sakazakii</Emphasis>) isolated from <Emphasis Type="Italic">Sunshik</Emphasis>, its ingredients and soils

The prevalence, genetic diversity, and antibiotic susceptibility of Cronobacter species (Enterobacter sakazakii) isolated from sunshik products, its ingredients, and root vegetable farm’s soils were investigated to analyze main reservoirs and contaminated sources of Cronobacter spp. Cronobacter spp. was isolated from 9 of 15 sunshik products, 26 of 72 its ingredients, and 2 of 39 soils. The root vegetables such as sweet potato and carrot showed higher contamination rate (70%) than the other sunshik ingredients. All isolates showed 929 bp band amplified from 16S rRNA and resistant to ampicillin, amoxicillin/clavulanic acid, and cefazolin. All isolates showed diverse random-amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE) band patterns. However, 3 cases of RAPD banding patterns between clinical strains and isolates from sunshik products and root vegetables (yam, carrot) were related with similarities level of 80%. These studies indicated that root vegetables can be an important contamination source of Cronobacter spp. in sunshik products. Thus, the preparation of root vegetables for manufacturing sunshik products used as a weaning diet was handled with more care than the other sunshik ingredients.     

Protective effect of carvacrol from <Emphasis Type="Italic">Thymus quinquecostatus</Emphasis> Celak against <Emphasis Type="Italic">tert</Emphasis>-butyl hydroperoxide-induced oxidative damage in Chang cells

Carvacrol is a monoterpenic phenol present in Thymus quinquecostatus Celak. The protective effects of carvacrol against tert-butyl hydroperoxide (t-BHP)-induced oxidative damage were investigated in human Chang cells. Cells treated with carvacrol extracts promoted Chang cell survival and protection was associated with stabilization of the mitochondrial membrane potential (MMP), prevention of oxidative stress-triggered reactive oxygen species (ROS), and lipid peroxidation (MDA production). In addition, Annexin V/PI, observed using Hoechst staining, indicated that carvacrol inhibited t-BHP-induced cell damage and stimulated the antioxidant capability of Chang cells due to elevation of glutathione (GSH) levels, which were reduced by t-BHP treatment. Carvacrol prevented oxidative stress-induced Chang cell damage via suppression of ROS and MDA levels, and increased GSH levels.     

Effect of <Emphasis Type="Italic">Lactobacillus acidophilus</Emphasis> Bauer and <Emphasis Type="Italic">Bifidobacterium animalis</Emphasis> ssp. <Emphasis Type="Italic">lactis</Emphasis> BB12 on proteolytic changes in dry-cured loins

The effect of the potentially probiotic bacteria strain of Lactobacillus acidophilus Bauer and probiotic bacteria Bifidobacterium animalis ssp. lactis BB12 on proteolytic changes of proteins in dry-cured loins during fermentation and cold storage was studied. Results of the conducted tests demonstrated that the use of probiotic bacteria for the production of dry-cured meats impacts the generation of products of protein proteolysis with high antioxidant activity. The highest antioxidant activity of peptides after fermentation and cold storage was observed in the loin with the strain B. animalis ssp. lactis BB12 and the loin with the mixture of strains L. acidophilus Bauer and B. animalis ssp. lactis BB12. Qualitative analysis of peptides demonstrated that peptides with weight below 3.5 kDa are characterized by the highest capacity of quenching the ABTS cation radical, including the peptides in loins with the strain B. animalis ssp. lactis BB12.     

Effect of heat-treated <Emphasis Type="Italic">Nuruk</Emphasis> on the quality characteristics of aged <Emphasis Type="Italic">Yakju</Emphasis>

Long-term aging of Yakju, a traditional Korean liquor made of rice and Nuruk (a fermentation agent), causes browning and odor and flavor development. This study investigated the effects of heat-treated Nuruk (50–80 °C, 30 min) on Yakju quality. The saccharogenic powers and glucoamylase, α-amylase, and carboxypeptidase activities were similar in non-heat-treated Nuruk and that treated at 50 °C. However, acidic protease and alcohol dehydrogenase decreased above 50 °C. The content of nitrogen-containing compounds was inversely proportional to the heat-treatment temperature. Compounds that cause off-flavors decreased at 50–60 °C, but increased at 70–80 °C, whereas compounds that provide fragrance increased at 50–60 °C. Sensory evaluation indicated that bad taste attributes were higher in Yakju produced using non-heat-treated Nuruk. Therefore, heat treatment of Nuruk at 50 °C can be adopted as a method for improving Yakju quality, as enzymatic activities that affect color, aroma, and taste are regulated.     

Differences of Bactericidal Efficacy on <Emphasis Type="Italic">Escherichia coli</Emphasis>, <Emphasis Type="Italic">Staphylococcus aureus</Emphasis>, and <Emphasis Type="Italic">Bacillus subtilis</Emphasis> of Slightly and Strongly Acidic Electrolyzed Water

In the present study, the disinfection efficacy of slightly acidic electrolyzed water (SAEW) and strongly acidic electrolyzed water (AEW) was tested on three bacteria, Escherichia coli, Staphylococcus aureus, and Bacillus subtilis and the disinfection mechanism was discussed. The results showed that SAEW had a stronger antibacterial efficacy against these tested bacteria in comparison with AEW. The results also showed that both SAEW and AEW treatments could damage the cell membrane, which was demonstrated microcosmically by scanning electron microscopy (SEM), thus causing leakages of protein, DNA, RNA, and ATP, resulting in the death of microbes. Moreover, AEW treatment could not cause the degradations of DNA and RNA, and nucleic acids including DNA and RNA are not the target point of its bactericidal efficacy. However, SAEW could maybe cause the degradation of RNA, and RNA may be the target in its antibacterial activity. We suggested that the differences in antibacterial efficacy between SAEW and AEW could be explained by the different impacts on RNA of tested strains.