Assessment of GH1, CAPN1 and CAST polymorphisms as markers of carcass and meat traits in Bos indicus and Bos taurus–Bos indicus cross beef cattle

The objective of this study was to investigate the effects of bovine GH1, CAPN1 and CAST gene polymorphisms on carcass and meat traits in Nellore and Nellore x Bos taurus beef cattle. Three hundred animals were genotyped for GH1/MspI (TC/G in intron 3), CAPN316 (AF_252504.2:g.5709C > G) and CAST/RsaI (AY_008267.1:g282C > G) and phenotyped for rib eye area, backfat thickness, intramuscular fat, shear force (SF), and myofibrillar fragmentation index (MFI). No significant associations were observed between the GH1/MspI and CAST/RsaI polymorphisms and phenotypes, although the relation between the CAST/RsaI genotypes and meat tenderness evaluated by MFI approached significant. The fact that the CAPN316 polymorphism did not show adequate segregation in Nellore cattle confirms the difficulty of using this marker in breeding programs of different Bos indicus breeds. However, the positive results of the association analysis obtained for Nellore x B. taurus crosses contributed to the validation of previous findings.     

Volatiles of the Balkan endemic Daucus guttatus ssp. zahariadii and cultivated and wild-growing D. carota – A comparison study

The hydrodistilled essential oil obtained from the aerial parts of Daucus guttatus Sibth. & Sm. ssp. zahariadii Heywood, an endemic plant species of the Balkan Peninsula, as well as solvent extracts and essential oils from different parts of Daucus carota L. have been analysed by GC and GC–MS and screened for antimicrobial activity against 12 bacterial and two fungal strains. The volatiles of the two plant taxa differed significantly in both their chemical identity and antimicrobial effect. The dominant constituent of D. guttatus oil was apiol (43.3%), which was absent from all samples of D. carota. The diethyl ether extract of D. carota inhibited the growth of the yeast Candida albicans while the oil of D. guttatus at 25 mg/ml had no effect on the growth of the fungal organisms tested. Additionally, the oil of D. guttatus showed prominent antibacterial activity against a pathogenic Corynebacterium pyogenes.     

Characterization of Aspergillus oryzae glycoside hydrolase family 43 β-xylosidase expressed in Escherichia coli

This is the first report of glycoside hydrolase family 43 β-xylosidase from Aspergillus oryzae. To characterize this enzyme, the recombinant enzyme was expressed in Escherichia coli. Unlike known β-xylosidases from fungal origins, the enzyme did not show substrate ambiguity and was stable at alkaline pH.     

Extracellular secretion in Bacillus subtilis of a cytoplasmic thermostable β-galactosidase from Geobacillus stearothermophilus

β-Galactosidase catalyzes the hydrolysis of β-galactosides into monosaccharides and is widely used in dairy processing. This study reports the extracellular secretion of a cytoplasmic thermostable β-galactosidase from Geobacillus stearothermophilus IAM11001 in Bacillus subtilis. This enzyme has potential applications in the dairy industry. It was not secreted in B. subtilis by mediation of 3 general secretory signal peptides, but was secreted extracellularly when it was fused to a twin-arginine signal peptide of B. subtilis phosphodiesterase. Defined and rich culture media were used for recombinant enzyme production, and the extracellular target enzymatic activity reached about 44% of the total enzymatic activity synthesized at 18 h of cultivation in Luria-Bertani medium. As a control of secretion, when the signal peptide coding sequence was absent from the N terminus of the target gene bgaB, the extracellular target enzymatic activity obtained under the same condition of cultivation accounted for less than 7% of the total enzymatic activity synthesized. Results also showed that coexpression of the B. subtilis proteins TatAd and TatCd was indispensable for the secretion of the target enzyme.     

Structure–activity relationships of acetylated flavone glycosides from Galeopsis ladanum L. (Lamiaceae)

Antioxidant activity, neuroprotective effect and acetylcholinesterase inhibitor activity were studied in acetylated flavones from Galeopsis ladanum L. (Lamiaceae), previously isolated and identified by UV-spectra, UPLC–MS/MS and ¹H-NMR spectroscopy. Structure–activity relationships of flavonoids have been determined in many antioxidant assays, generally focused on hydroxyl groups. In this study we have detected new interesting structure–activity relationships for the isolated flavonoids due to the acetylation of sugar moiety of these flavonoids. Methylation at 4′-OH and monoacetylation is beneficial to inhibiting AChE and shows a neouroprotective effect.     

Antioxidant properties of 1,2,3,4,6-penta-O-galloyl-β-d-glucose from Elaeocarpus sylvestris var. ellipticus

The antioxidant potential of 1,2,3,4,6-penta-O-galloyl-β-d-glucose (PGG), isolated from Elaeocarpus sylvestris var. ellipticus, was investigated by various established systems based on cell-free and cell system experiments, such as radical detection, antioxidant enzyme assay, lipid peroxidation detection, and cell viability assay. PGG was found to quench the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and intracellular reactive oxygen species. PGG recovered the cellular antioxidant enzyme activities of superoxide dismutase, catalase, and glutathione peroxidase, which were reduced by H₂O₂ treatment, thereby resulting in the inhibition of lipid peroxidation. Cytoprotective effects of PGG were based on the results of DNA fragmentation, mitochondrial membrane potential (Δψ), apoptotic body formation, and caspase-3 activity. The results suggest that PGG protects cells against H₂O₂-induced cell damage via antioxidant properties.     

In vitro and in vivo assessment of cytochrome P450-mediated herb–drug interaction of Ssang-hwa-tang

We have evaluated the herb–drug interaction potential of Ssang-hwa-tang (SHT) mediated by cytochrome P450 (CYP) inhibition/induction. Further, the effects of fermentation on the CYP-mediated herb–drug interaction potential were determined. SHT showed inhibitory activity toward CYP1A2, but not 2A6, 2B6, 2C9, 2C19, 2D6, 2E1, and 3A4 in human liver microsomes. The results of the enzyme kinetic study suggested that the SHT-induced CYP1A2 inhibition is mixed reversible inhibition. The hepatic CYP expression and activity in rats treated with SHT were examined. The expression/activity of CYP2E1 increased as a result of SHT extract treatment (P < 0.005 or P < 0.001, respectively), which raises the possibility that SHT may increase the toxicity of environmental toxicants through the elevation of CYP2E1-mediated metabolic activation. SHT fermentation using Lactobacillus fermentum or Lactobacillus gasseri resulted in attenuation of the SHT-induced CYP1A2 inhibition, but not CYP2E1 induction, suggesting that changes in the chemical composition of SHT through fermentation can affect the inhibition of CYP1A2 activity.     

Comparing the in vitro effects of MGO™ Manuka honey and tea tree oil on ocular Demodex viability

Purpose

To compare the in vitro antiparasitic effects of MGO^? Manuka honey and tea tree oil against ocular Demodex.

Acidification,crushing and thermal treatments can influence the profile and stability of folate poly-γ-glutamates in broccoli (Brassica oleracea L. var. italica)

The influence of different treatments, i.e., crushing, high temperature short time (90 °C/4 min) (HTST) and low temperature long time (60 °C/40 min) (LTLT) blanching, acidification (pH 4.3), and sequences of these treatments on the folate poly-γ-glutamate profile and stability were investigated. In this study, broccoli was used as a case study. Regarding the folate poly-γ-glutamate profile, endogenous folate poly-γ-glutamates in broccoli florets were found predominantly as hepta- and hexa-γ-glutamates. Crushing raw broccoli, acidification and LTLT blanching enhanced folate deconjugation resulting in monoglutamate, di- and tri-γ-glutamates. Compared to other treatments, HTST blanching preformed prior to crushing resulted in the highest concentration of long chain poly-γ-glutamates. Regarding folate poly-γ-glutamates stability, acidification combined with LTLT blanching decreased folate stability whereas HTST blanching combined with different sequences of blanching and crushing did not affect folate poly-γ-glutamates stability. It was concluded that crushing (prior to heating), acidification and blanching could be strategically applied to increase the folate monoglutamate content of broccoli.     

Novel α-glucosidase inhibitors from Macaranga tanarius leaves

One of the hyperglycaemic remedies is glucose absorption reduction by suppressing carbohydrate digestion due to utilisation of α-glucosidase inhibitors (AGIs). Determination of prospecting herbs done in vitro by using enzyme assay resulted in the finding of Macaranga tanarius, which showed a potent inhibitory activity. An EtOAc-soluble extract of M. tanarius leaves was chromatographed by a Diaion HP-20 column and the active fractions were further purified with high performance liquid chromatography (HPLC) to isolate active principles against α-glucosidase. Five ellagitannins were successfully isolated and identified. Structure determination revealed that these isolated compounds were mallotinic acid (IC₅₀ > 5.00 mM), corilagin (IC₅₀ = 2.63 mM), chebulagic acid (IC₅₀ = 1.00 mM), and two novel compounds named macatannins A (IC₅₀ = 0.80 mM) and B (IC₅₀ = 0.55 mM). AGIs play an important role for the treatment of diabetes, therefore this research results may suggest novel alternatives for diabetes treatment management.     

Short communication: Effect of αS1-casein (CSN1S1) and κ-casein (CSN3) genotypes on milk composition in Murciano-Granadina goats

The effects of the caprine α_S1-casein (CSN1S1) polymorphisms on milk quality have been widely demonstrated. However, much less is known about the consequences of the κ-casein (CSN3) genotype on milk composition in goats. Moreover, the occurrence of interactions between CSN3 and CSN1S1 genotypes has not been investigated. In this study, an association analysis between CSN1S1 and CSN3 genotypes and milk quality traits was performed in 89 Murciano-Granadina goats. Total milk yield as well as total protein, fat, solids-not-fat, lactose, α_S1-casein (CSN1S1), and α_S2-casein (CSN1S2) contents were recorded every other month during a whole lactation (316 observations). Data analysis using a linear mixed model for repeated observations revealed no interaction between the CSN1S1 and CSN3 genotypes. With regard to the effect of the CSN3 locus, AB and BB genotypes were significantly associated with higher levels of total casein and protein content compared with the AA CSN3 genotype. In strong contrast with French breeds, the CSN1S1 genotype did not affect protein, casein, and fat concentrations in Murciano-Granadina goats. These results highlight the importance of taking into consideration the CSN3 genotype when performing selection for milk composition in dairy goats.     

Enzymatic synthesis of fructooligosaccharides by inulinases from Aspergillus niger and Kluyveromyces marxianus NRRL Y-7571 in aqueous–organic medium

This work is focused on the synthesis of the fructooligosaccharides (FOS) from sucrose and inulin, using free, immobilized and pre-treated immobilized inulinase from Kluyveromyces marxianus NRRL Y 7571 and Aspergillus niger in an aqueous–organic system. Initially, the influence of pre-treatment using four different gases, propane, n-butane, CO₂ and liquefied petroleum gas (LPG), was investigated towards FOS production and best results were found when both enzymes were previously treated with LPG. The best reaction yields were obtained when the immobilized enzymes were treated with LPG. Considering FOS synthesis using the enzyme from A. niger, yields of 26.62% of GF2 (kestose), 30.62% of GF3 (nystose) and 8.47% of GF4 (fructosyl nystose) were achieved using sucrose as substrate. Using inulinases from K. marxianus NRRL Y 7571, 11.89% of GF2 and 20.83% of GF3 were obtained, using inulin as substrate. However, promising results were achieved using the free form of inulinase from A. niger (77.19% of GF2; 14.03% of GF3 and 0.07% of GF4) using inulin as substrate.     

α-Glucosidase inhibitors from Chinese Yam (Dioscoreaopposita Thunb.)

The inhibitory activities of crude extracts and purified constituents from the fresh tuberous rhizomes of Chinese Yam (Dioscorea opposita Thunb.), which is commonly called Huai Shan Yao in Chinese, were evaluated against yeast α-glucosidase in order to search for the active principals for treatment of diabetes. Bioassay-guiding isolation gave four compounds: trans-N-p-coumaroyltyramine (1) (IC₅₀ = 0.40 μM), 1,7-bis(4-hydroxyphenyl)heptane-3,5-diol (2) (IC₅₀ = 0.38 mM), 6-hydroxy-2,4,7-trimethoxyphenanthrene (3) (IC₅₀ = 0.77 mM) as α-glucosidase inhibitors, and cis-N-p-coumaroyltyramine (4), an isomer of compound 1, which showed no inhibitory activity against α-glucosidase. Furthermore, the separation and purification of compound 3 from Chinese Yam (Huai Shan Yao) was conducted by high-speed counter-current chromatography (HSCCC) using hexane–ethyl acetate–methanol–water (1:1:1:1, v/v/v/v). Compound 1, 2 and 4 were isolated from or detected in the Dioscoreaceae family for the first time.     

Survival of three Bifidobacterium animalis subsp. lactis strains is related to trans-vaccenic and α-linolenic acids contents in organic fermented milks

This study aims at relating the survival at 4 °C for 28 days in organic and conventional fermented milks of three strains of Bifidobacterium animalis subsp. lactis (BB12, B94 and BL04), in co-culture with Streptococcus thermophilus TA040 and Lactobacillus bulgaricus LB340, to milk fatty acids profile. Cultivability after 28 days of cold storage was improved in organic fermented milks as compared to conventional products, with slight differences among strains. In addition, the poly-unsaturated fatty acids fraction was higher in organic products, as well as the relative trans-vaccenic (TVA) and α-linolenic (ALA) fatty acids contents that were respectively 1.7 and 2.4 times higher in organic than in conventional fermented milks. From these results, it was concluded that elevated levels of TVA and ALA, together with a lower ratio between linoleic acid and α-linolenic acid, as found in organic products, improved the survival of the bifidobacteria during chilled storage.     

α-Glucosidase inhibitors from the bulb of Eleutherine americana

One effective way to treat diabetes is by suppressing carbohydrate digestion due to the utilisation of α-glucosidase inhibitors (AGIs). The determination of prospective herbs, done in vitro by using enzyme assay, resulted in the finding of Eleutherine americana, which showed a potent inhibitory activity. A 50% aqueous methanol-soluble extract of bulbs of E. americana was chromatographed successively and the active fractions were further purified with preparative high performance liquid chromatography (HPLC) to isolate active compounds against α-glucosidase. Structure determination by mass and NMR analysis revealed that these isolated compounds were eleutherol (1), eleutherinoside A (2), and eleuthoside B (3) based on comparisons with the reference data. Considering the amount and the inhibitory activity of each naphthalene in the whole extracts, the bulb of E. americana inhibitory activity against α-glucosidase might be a result of compound 2 (IC₅₀ = 0.5 mM, yield = 5 mg/50 g plant sample, with a characteristic structure which has never been found in other AGIs. AGIs play an important role for the treatment of diabetes, therefore these results may suggest novel alternatives for diabetes treatment management.     

In-vitro evaluation of hydrocolloid–based encapsulated fish oil

The release behaviour of hydrocolloid-based encapsulated fish oil was carried out to understand its applicability as a controlled release delivery system. A casein-based Maillard reaction product (MRP), [caseinate–glucose-dried glucose syrup, MRP] and its corresponding non-MRP-based encapsulant materials were used to form stable emulsions with fish oil. The emulsion particle size distribution was found to be uniform with 90% of the particles below 1.3 μm. These oil/water emulsions were spray dried to obtain free-flowing fish oil microcapsules. All casein-based microcapsules had a free fat content of <1% and the encapsulation efficiencies were all >97%. A sequential in-vitro release protocol and a method for evaluation of the released oil in simulated gastric and intestinal fluids (SGF/SIF) were developed and tested in this study. A comparative analysis of release behaviour between caseinate and whey protein-based MRP and non-MRP-based encapsulant materials was also obtained. The amount of released oil after subjecting to the sequential in-vitro protocol was found to be <2% and 36% for caseinate and whey protein-based MRP microcapsules respectively.     

A combination of TiO2–UV photocatalysis and high hydrostatic pressure to inactivate Bacillus cereus in freshly squeezed Angelica keiskei juice

High hydrostatic pressure (HHP) is an effective nonthermal food processing method for microbial inactivation with minimal change to sensory and nutritional values. However, the resistance of endospores to high pressure hinders application of HHP to freshly squeezed vegetable juice, especially from leafy vegetables that are susceptible to contamination of soil bacteria, such as endospore-forming Bacillus cereus. A combination of TiO₂–UV photocatalysis and HHP was used in the processing of freshly squeezed Angelica keiskei juice, and inactivation of naturally occurring microbes, especially B. cereus, was investigated. Yeasts and molds, coliform bacteria, and Pseudomonas were inactivated by HHP to levels below the detection limit, but B. cereus survived HHP and grew during 8 days of subsequent refrigerated storage (4 °C). However, no colonies of yeasts and molds, coliform bacteria, Pseudomonas, or B. cereus were detected in A. keiskei juice after processing with a combination of TiO₂–UV photocatalysis and HHP. Although B. cereus in juice processed with the combination treatment recovered and grew to 2.02 log CFU/mL on day 6 of storage, this level was less than the population of B. cereus in unprocessed Angelica keiskei juice immediately after squeezing.     

Determination of polyphenol levels variation in Capsicum annuum L. cv. Chelsea (yellow bell pepper) infected by anthracnose (Colletotrichum gloeosporioides) using liquid chromatography–tandem mass spectrometry

Healthy fruits of Capsicum annuum L. cv. Chelsea (yellow bell pepper) and one infected by Colletotrichum gloeosporioides were analysed for polyphenols via high-performance liquid chromatography coupled with tandem mass spectrometry (HPLC–MS/MS). Among seven polyphenols characterized, four components in the C. annuum fruits were identified for the first time. To investigate the characteristics of the polyphenols as defence materials, the content change of the fruit polyphenols inoculated with C. gloeosporioides was monitored by HPLC. It was observed for the first time that de novo induced N-caffeoyl putrescine (1) and caffeoyl O-hexoside (2) appeared to act as a phytoalexin in the defence mechanism of the C. annuum fruits against C. gloeosporioides, and constitutively formed feruloyl O-glucoside (3), kaempferol O-pentosyldihexoside (4) and dihydroxyflavone O-hexoside (7) as a phytoanticipin in the diseased C. annuum fruits.