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.     

Phenolic contents and antioxidant activities from different tissues of <Emphasis Type="Italic">Baekseohyang</Emphasis> (<Emphasis Type="Italic">Daphne kiusiana</Emphasis>)

The aim of this research was to investigate the levels of phenolic compounds and antioxidant activities in different tissues including leaves, stems, and roots from baekseohyang (Daphne kiusiana). The highest contents of total phenolics (43.59 mg gallic acid equivalent, GAE/g) and flavonoids (15.73 mg rutin equivalents, RE/g) were observed in the 75% methanol extract of leaves. Moreover, this extract had the predominant antioxidant capacity, DPPH (85.91%) and ABTS (92.57%) radical scavenging activities as well as reducing power (7.20%) at a concentration of 5 mg/mL. The highest content of phenolic compounds was also exhibited in this extract with an increasing order in leaves, roots, and stems and their major components were vanillic acid (6.37 mg/g), tannic acid (1.91 mg/g), and p-hydroxybenzoic acid (3.96 mg/g). Thus, the strong antioxidant activities of the 75% methanol extract are correlated with high phenolic compound contents. This study suggests that baekseohyang leaves may potentially be used as an accessible source of natural antioxidants.     

Antitumor activities of liposome-incorporated aqueous extracts of <Emphasis Type="Italic">Anodonta </Emphasis><Emphasis Type="Italic">woodiana</Emphasis> (Lea, 1834)

To acquire the effectiveness of oral treatments, aqueous extracts of Anodonta woodiana (HB) were incorporated into liposome and its antitumor activities evaluated in vivo. The HB-loaded liposomes (HBL) were prepared at a mean size of 14.85 μm by reverse-phase evaporation method. Referring to the maximum tolerated doses test, mice with orally administrated HBL, at a 3 g/kg body weight dosage, showed no obvious acute toxic sign. Furthermore, the tumoricidal activities of HBL against C26 murine colon carcinoma, Lewis lung tumor, human QGY hepatic carcinoma and MKN-45 gastric tumor were examined, respectively. In contrast to free HB, HBL possessed remarkable antitumor activity. Simultaneously, the effect of HBL was observed in a dose-dependent manner. For C26 murine colon carcinoma and Lewis lung tumor, the inhibitory ratios of HBL were 54.36 and 51.97% at a dose of 400 mg/kg, respectively. The suppression of tumor growth and the reduction in body weight were more pronounced in human QGY hepatic carcinoma and MKN-45 gastric tumors inoculated mice by treatment of HBL during 30 days. All these promising results implied that liposome-incorporated aqueous extracts of Anodonta woodiana had a more potential application as a natural antitumor and immunomodulator formulation.     

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.     

<Emphasis Type="Italic">Aronia melanocarpa</Emphasis> phenolics and their antioxidant activity

The purpose of the present study was to evaluate small and high molecular phenolics (tannins) and antioxidant activity of Aronia melanocarpa berries, juice and pomace in order to find new potential sources of natural antioxidants. The fruits of Aronia melanocarpa Elliot were collected in the middle of October at a plantation near Wroclaw, Poland. The pomace has a much higher content of phenolics in comparision to juice and fruits. Results showed that polymeric proanthocyanins, predominantly of (−)epicatechin, are the major class of polyphenolic compounds in chokeberry, represent 66% of fruits polyphenols. The average concentration ranged from 1578.79 mg/100 g of DW for chokeberry juice up to 8191.58 mg/100 g in pomace. The concentration of phenolic acids (chlorogenic and neochlorogenic acids) in juice was higher than in pomace. Anthocyanins in Aronia melanocarpa are second phenolic compound group and represent about 25% of total polyphenols, mixture of four different cyanidin glycosides: 3-galactoside, 3-glucoside, 3-arabinoside and 3-xyloside. The higher antioxidant activity expressed as TEAC was measured in pomace >fruit >juice.     

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.     

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.     

Pharmacognostic standardization and antioxidant capacity of an edible mushroom <Emphasis Type="Italic">Laetiporus sulphureus</Emphasis>

Traditionally Laetiporus sulphureus, commonly known as “Chicken-of-the-Woods”, is considered a delicacy in various parts of the world and recently has shown medical potential in several treatments such as antitumor, antimicrobial, anti-inflammatory, antioxidant, immune-modulating etc. However, quality standards of this species have not been studied so far. For this purpose, microscopic features of powder such as physical characters, spore measurement, type of hyphae were examined and recorded. Physicochemical parameters like organoleptic features, loss on drying (5.15 %), total ash (3.04 %), swelling capacity (1.66 ml/g), water holding capacity (163.16 g/g), water solubility index (21.73 %) and fluorescent behaviour against 16 reagents were also determined. In addition, alcoholic extract was prepared using methanol as solvent where extractive value was 24.27 %. Preliminary mycochemical analysis indicated presence of cardiac glycoside, flavonoid, phenol, saponin and terpenoids in the extract. While quantitatively major bioactive components were present in the following order of flavonoid > phenol > ascorbic acid > β-carotene > lycopene. Moreover, HPTLC and HPLC profiles were recorded to determine phenolic fingerprint. HPTLC characterization showed presence of at least 5 components as detected by scanning the plate at 300 nm and HPLC chromatogram indicated existence of 32 peaks which might be of phenolic compounds. In addition, superoxide radical scavenging (EC₅₀ 1.38 mg/ml), DPPH radical scavenging (EC₅₀ 0.11 mg/ml), chelating ability of ferrous ion (EC₅₀ 0.27 mg/ml), reducing power (EC₅₀ 0.26 mg/ml) and total antioxidant capacity (18.01 µg AAE/mg of extract) were determined to evaluate antioxidant potentiality of the extract. The above parameters are significant towards establishing pharmacognostic standards for future authentication of genuine mushroom material.     

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.     

Quantification of anthocyanins in black carrot extracts (<Emphasis Type="Italic">Daucus carota</Emphasis> ssp.<Emphasis Type="Italic"> sativus</Emphasis> var.<Emphasis Type="Italic"> atrorubens</Emphasis> Alef.) and evaluation of their color properties

Pigment composition of 15 black carrot cultivars (Daucus carota ssp. sativus var. atrorubens Alef.) was screened by HPLC-MS. Up to seven cyanidin glycosides, five of which were acylated with hydroxycinnamic and hydroxybenzoic acids, were identified and quantified in the roots by HPLC-DAD. Contents of individual compounds indicated great differences in the potential of anthocyanin accumulation both between different cultivars and carrots of the same cultivar. Total anthocyanin amounts ranged from 45.4 mg/kg dry matter to 17.4 g/kg dry matter. To the best of our knowledge, this is the first report on the quantification of individual anthocyanins in roots of different black carrot cultivars. The determination of color properties in the extracts under various pH conditions proved black carrot anthocyanins to be applicable as natural food colorants also for low-acid food commodities, whereas a considerable loss of color was noted under nearly neutral conditions. Additionally, relatively high saccharide contents were found in almost all cultivars which may be disadvantageous when coloring concentrates are produced from carrot roots.     

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.     

Antioxidant and anticholinesterase activity of essential oils and ethanol extracts of <Emphasis Type="Italic">Thymus algeriensis</Emphasis> and <Emphasis Type="Italic">Teucrium polium</Emphasis> from Algeria

This study aimed to investigate the cholinesterase (ChE) inhibitory and antioxidant activities of essential oil (EO) and ethanolic extract (EE) of two Lamiaceae medicinal plants from Algeria: Thymus algeriensis Boiss. & Reut. and Teucrieum polium subsp capitatum. The chemical composition of EOs analyzed by GC and GC–MS revealed the presence of carvacrol (43.2%), p-cymene (18.7%) and γ-terpinene (14.8%) as major compounds in T. algeriensis, while germacrene D (25.0%), bicyclogermacrene (10.4%), β-pinene (11.3%) and spathulenol (5.8%) were the most important components in T. polium oil. The best inhibitory activity on acetylcholinesterase (AChE) was exhibited by the EO of T. algeriensis. On the other hand, T. polium oil was more efficient against butyrylcholinesterase (BChE) than against AChE, whereas EEs showed weak or no inhibitory effect, particularly against AChE. Antioxidant activity was evaluated by β-carotene bleaching, ferric and cupric reducing powers and inhibition of ABTS^?+, DPPH^? and superoxide radicals. EOs showed different antioxidant trends depending on the assay used while ethanol extracts had high activity with all tests.     

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.     

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.     

Exposure assessment of <Emphasis Type="Italic">Staphylococcus aureus</Emphasis> in <Emphasis Type="Italic">Kimbab</Emphasis>, a ready-to-eat Korean food

The exposure assessment was carried out for Staphylococcus aureus in kimbab by predicting growth of S. aureus and the production of enterotoxin using Food MicroModel® program. Environmental parameters selected were pH 5.5, Aw 0.999, and storage temperatures in the range of 10 to 30°C. It was predicted that 6.3 hr could be a critical time for enterotoxin production while kimbab was stored at 30°C. Mild case scenario analysis showed that enterotoxin could not be produced if kimbab was kept at 10°C during preparation and distribution and then left at 25°C for 4 hr before consumption. In the worst case scenario, the keeping time at 25°C was assumed to be 7.0 hr. The level of S. aureus in the worst case was predicted to be 6.8×10⁶ CFU/g which is lower than the critical level (7.8×10⁶ CFU/g) for toxin production.     

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.     

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.