Metagenomic and volatile profiles of ripened cheese obtained from dairy ewes fed a dietary hemp seed supplementation.

Chemical and organoleptic properties of dairy products largely depend on the action of microorganisms that tend to be selected in cheese during ripening in response to the availability of specific substrates. The aim of this work was to evaluate the effects of a diet enriched with hemp seeds on the microbiota composition of fresh and ripened cheese produced from milk of lactating ewes. Thirty-two half-bred ewes were involved in the study, in which half (control group) received a standard diet, and the other half (experimental group) took a diet enriched with 5% hemp seeds (on a DM basis) for 35 d. The dietary supplementation significantly increased the lactose in milk, but no variations in total fat, proteins, caseins, and urea were observed. Likewise, no changes in total fat, proteins, or ash were detected in the derived cheeses. The metagenomic approach was used to characterize the microbiota of raw milk and cheese. The phyla Proteobacteria and Firmicutes were in equally high abundance in both control and experimental raw milk samples, whereas Bacteroidetes was less abundant. The scenario changed when considering the dairy products. In all cheese samples, Firmicutes was clearly predominant, with Streptococcaceae being the most abundant family in the experimental group. The reduction of taxa observed during ripening was in accordance with the increment (relative abundance) of the starter culture Lactococcus lactis and Streptococcus thermophilus, which together dominate the microbial community. The analysis of the volatile profile in ripened cheeses led to the identification of 3 major classes of compounds: free fatty acids, ketones, and aldehydes, which indicate a prevalence of lipolysis compared with the other biochemical mechanisms that characterize the cheese ripening.

Analysis of power from palm oil solid waste for biomass power plants: A case study in Aceh Province.

Nowadays, the solid waste produced from palm oil has become one of the essential oils in the world in general and especially in Indonesia. Biomass waste is processed through substantial quantities of palm oil extraction. With the reduction in fossil fuels in recent years, it has had an impact on the deterioration of electricity supply at the National and International levels. Biomass is a renewable energy that can replace conventional energy. Besides, power generation from biomass is environmentally friendly and sustainable. This simulation was conducted to analyze the maximum power from the burning of oil palm biomass for the electricity generation. The novelty of the article is the performance and behavior of palm oil biomass-based co-fuel in the power generation process. The biomass wastes used in this simulation include OPF, EFB, PKS, and OPM. The results of this simulation indicate that the maximum power produced with OPF can produce 49.54 MW with variations in the flow rate of biomass at 8 kg/s. While at the time of recycling up to 100% OPM biomass produces 61.05 MW higher than OPF, EFB and PKS. Meanwhile, the OPF-PKS mixed biomass can produce 106.15 MW of power plants when the airflow rate reaches 171 kg/s. The overall results of the simulation for the analysis of the maximum power that can be used as a power station show suitability and can apply in rural/remote areas. Besides, the availability of oil palm biomass in Aceh Province is also sufficient to overcome electricity shortages and reduce dependence on conventional energy.

Ethnomycological knowledge among Kaqchikel, indigenous Maya people of Guatemalan Highlands.

BACKGROUND: The Guatemalan Highlands is a region of great but so far poorly known mycological diversity. People living in this area have long used wild fungi as a source of food and income. However, our knowledge of the ethnomycological practices of the Mayan peoples of Guatemala is still rudimental, especially if compared with information reported for the neighboring region of Mexico. Among the main indigenous groups of the Maya people inhabiting the highlands of Central Guatemala, stand the Kaqchikel, accounting for nearly 8% of the entire Guatemalan population. The main aim of this study was to record the traditional knowledge and use of edible wild mushrooms by inhabitants of the municipality of San Juan Sacatepéquez that lies at the heart of the Kaqchikel area in the central highlands of Guatemala, also describing the relevant selling practices and dynamics. A secondary aim was to compare the diversity and composition of the mushroom assemblage offered at the market with the macrofungal diversity of woods in the area. METHODOLOGY: This study is the result of 4 years of ethnomycological research, conducted through continuous visits to the municipal market and focused interviews with collectors and vendors. Field sampling in pine-oak forested areas surrounding San Juan Sacatepéquez, from where the mushrooms sold at the market are foraged, were also conducted, in the presence of local collectors. RESULTS: The results show a significant richness of species sold in the market, a network of commerce of purchase, sale, and resale of several species, with relatively stable prices, and knowledge about edible and inedible species that is transmitted mainly within the family nucleus. The business of selling mushrooms in the market is an exclusive activity of women, who are supplied by collectors or by other vendors. Fungi are sold and bought only as food, while no consumption of hallucinogenic mushrooms or medicinal mushrooms was recorded. Several species of Amanita, Cantharellus, Boletus, Lactarius, and Russula were those most commercialized in the 4 years of the study, but we also spotted fungi never reported before as consumed in the country, including Gastropila aff. fumosa (= Calvatia fumosa) and several species of Cortinarius. Field sampling in nearby pine-oak forests confirmed an elevated local macrofungal diversity. CONCLUSION: Our study unveiled the contemporary wealth of Kaqchikel culture for what concerns mushrooms, demonstrating that mushrooms continue to be culturally and economically important for these communities despite the erosion of traditional knowledge. Our results also confirmed the need to investigate in greater detail the Guatemalan mycodiversity that is vast and poorly known.

The educational intervention "Focusing" as a strategy to stress reduction among health care workers: a pilot study in an Italian teaching hospital.

BACKGROUND: An educational intervention for stress management of healthcare workers based on a mindfulness approach called Focusing in an Italian teaching hospital. METHODS: In 2017 a pilot mandatory training on stress management targeted to healthcare workers was organized in the 447 bed teaching hospital Sant'Andrea in Rome. We implemented a mindfulness intervention called Focusing. Training with an adult learning approach was adopted. A mixed model using quantitative and qualitative methods was used to evaluate the course's effects on stress reduction. Quantitative data were collected with the Perceived Stress Scale and data were analyzed at two different time points: pre-intervention and post-intervention. Qualitative data were collected via a focus group interview at the end of the course. RESULTS: A total of 20 healthcare workers participated in the course and was divided in 2 groups: 7 physicians and 13 nurses. 15 (75%) were women and ages ranged from 31 to 56 (median age 46,5 years). 5 (25%) were men and ages ranged from 39 to 54 (median age 47 years). Perceived stress decreased with significant change (P=,0195) from 21,4 +/- 4,4 at baseline to 17,5 +/- 6.18 at the end of the course. The major changes the participants noted in their focus group were a greater sense of calmness, enhanced emotional self-regulation, improved coping, increased mental clarity and a sense of empowerment in dealing with the fear, anxiety, and other issues related to their work. CONCLUSIONS: Our initial aim was to investigate whether the course, based on improve of self-empathy by Focusing, could help healthcare workers face the negative effects of stress. There was a significant reduction in perceived stress and qualitative results showed an improvement in reactivity to inner experience and a more attentive perception of internal and external experiences.

A preliminary characterization of Aglianico (Vitis vinifera L. cv.) grape proanthocyanidins and evaluation of their reactivity towards salivary proteins.

The flavan-3-ol and proanthocyanidin composition of Aglianico seeds and skins were for the first time determined by HPLC-MS in comparison with the international grapes Merlot and Cabernet Sauvignon. Monomers [(+)-catechin C, (-)-epicatechin EC, (-)-epicatechin-3-O-gallate, ECG] and oligomers [B1, B2, B3, B4 dimers and trimer C1] were identified and quantified in grape extracts. In order to evaluate the reactivity towards salivary proteins of model wine solutions of seeds and skins monomeric/oligomeric and polymeric fractions, the Saliva Precipitation Index (SPI) was carried out. Fractions were also analyzed for their mean degree of polymerization (mDP), percentage of galloylation (%G) and of prodelphinidin (%P) by phloroglucinolysis. Aglianico was the most effective in precipitating proteins than Merlot and Cabernet Sauvignon, mainly for the high percentage of galloylation of grape fractions. The mDP and the percentage of ECG in terminal units resulted to significantly contribute to the precipitation of salivary proteins by grape proanthocyanidins.

Catalase and antiquitin from Euphorbia characias: two proteins involved in plant defense?

Here we report the cDNA nucleotide sequences of a calmodulin-binding catalase and an antiquitin from the latex of the Mediterranean shrub Euphorbia characias. Present findings suggest that catalase and antiquitin might represent additional nodes in the Euphorbia defense systems, and a multi-enzymatic interaction contributing to plant's protection against biotic and abiotic stresses is proposed to occur in E. characias laticifers.

Olive milling wastewater as a medium for growth of four Pleurotus species.

Four species of Pleurotus were adapted to grow on olive milling wastewater, and in certain conditions produced high yield of fruit bodies. Some biochemical transformations were observed in the olive milling wastewater owing to the growth of Pleurotus. In particular, the fungi actively excreted large amounts of laccase in the medium, and at the same time the concentration of phenolics and other toxic compounds significantly decreased, as revealed by HPLC analysis and toxicity tests on standard cultures of human cell lines.

DNA methylase from Pisum sativum.

DNA methylase activity was detected in nuclei from pea shoots. The enzyme can only be extracted by low-salt treatment if the nuclei are pretreated with micrococcal nuclease. Only a single enzyme was detected, and it was purified to a specific activity of 1620 units/mg of protein. It has an Mr of 160,000 on gel filtration and SDS/PAGE. Pea DNA methylase methylates cytosine in all four dinucleotides, and this is interpreted to show that it acts on CNG trinucleotides. Although it shows a strong preference for hemi-methylated double-stranded DNA, it is also capable of methylation de novo. Homologous DNA is the best natural substrate. In vitro the enzyme interacts with DNA to form a salt-resistant complex with DNA that is stable for at least 4 h.

Eukaryotic DNA methylases and their use for in vitro methylation.

DNA methylases from mouse and pea have been purified and characterized. Both are high molecular mass enzymes that show greater activity with hemimethylated than unmethylated substrate DNA. Both methylate cytosines in CpG preferentially, but not exclusively and show similar kinetics of methylation, which makes it difficult to saturate all possible sites on the DNA, but procedures are described that circumvent this problem.

Amine oxidase from Lathyrus cicera and Phaseolus vulgaris: purification and properties.

Cu-Amine oxidases (amine oxygen oxidoreductase deaminating, copper containing E.C. 1.4.3.6.) are found in all forms of life (1). They catalyze the following general reaction: R-CH2-NH2 + O2 + H2O----R-CHO + NH3 + H2O2. Cu-amine oxidases (Cu-AOs) have been extracted from different leguminosae: Pisum sativum (2-3), Lathyrus sativus (4), Lens esculenta (5), Vicia faba (6), Cicer arietinum (7), Glycine max (8) but not from Phaseolus vulgaris. Palavan and Galston (9), in a study of polyamine biosynthesis during developmental stages of Phaseolus vulgaris, did not detect diamine or polyamine oxidase activity in Phaseolus. The present paper describes the purification of Phaseolus vulgaris seedlings amine oxidase (PhSAO) and also compares the properties of this enzyme to the Lathyrus cicera enzyme (LcSAO), obtained with the same method of purification.

Microheterogeneity of lentil seedling (Lens culinaris) amine oxidase.

Purified lentil seedling amino oxidase (LSAO) is homogeneous in the analytical ultracentrifuge, but shows heterogeneity in gel-filtration HPLC and in PAGE. Two components were obtained from HPLC and PAGE, but at least 6-7 subforms were seen by electrofocusing techniques. The chromatographic and electrophoretic forms are not interconvertible, indicating the presence of covalent differences. The electrophoretic pattern, but not the chromatographic pattern, is modified by treating the enzyme with a pool of glycohydrolases. The copper-free enzyme shows the same type of heterogeneity as the native enzyme, this ruling out the possibility that some subforms were due to the presence of apoenzyme.

Transient kinetics of copper-containing lentil (Lens culinaris) seedling amine oxidase.

The reaction between lentil (Lens culinaris) seedling amine oxidase and its chromogenic substrate, p-dimethylaminomethylbenzylamine, has been studied by the stopped-flow technique. Upon being mixed with substrate in the absence of oxygen, the enzyme is bleached in a complex kinetic process. A yellow intermediate absorbing at 464 nm and the first product (aldehyde) are formed in subsequent steps. When oxygenated buffer is mixed with substrate-reduced amine oxidase, the 496 nm absorption of the oxidized enzyme is very rapidly restored in a second-order process (k = 2.5 X 10(7) M-1 X S-1). This reaction is appreciable even at very low oxygen concentration, in keeping with the fairly low Km for O2 measured by steady-state kinetics.

Lentil seedlings amine oxidase: preparation and properties of the copper-free enzyme.

The reaction of copper-free lentil seedlings amine oxidase with substrates has been studied. While devoid of catalytic activity, this enzyme preparation is still able to oxidize two moles of substrate and to release two moles of aldehyde and two moles of ammonia per mole of dimeric protein. The same stoichiometry has been determined on the native enzyme in the absence of oxygen. Although copper is essential for the reoxidation of the reduced enzyme, a binding of oxygen to the copper-free protein has been demonstrated.

Reaction of beef plasma and lentil seedlings Cu-amine oxidases with phenylhydrazine.

The reaction of copper amine oxidases from beef plasma and lentil seedlings with hydrazine derivatives has been studied. A 1:1 stoichiometry was always found for the irreversible binding to the dimeric proteins. The formation of the adduct does not require the presence of oxygen or copper. Substrates compete with hydrazine derivatives for the binding to the enzymes. The binding of hydrazines and of substrate has different effects on the EPR spectra of enzymic copper.

Synthesis and chromatographic properties of Se-carboxymethyl-selenohomocysteamine (2-carboxymethyl, 3-aminopropyl selenide).

Details are reported for the synthesis of Se-carboxymethylselenohomocysteamine from selenohomocysteamine and monochloroacetic acid. Data on its behaviour on paper and ion-exchange chromatography are also reported, which allow its identification.

Synthesis and chromatographic properties of selenazolidine-4-carboxylic acid (selenaproline).

Details are reported for the synthesis of DL-selenazolidine-4-carboxylic acid starting from DL-selenocystine and formaldehyde. Some chemical reactions and the paper and ion-exchange chromatographic behaviour of selenazolidine carboxylic acid in comparison with thiazolidine-4-carboxylic acid are described.

Chromatographic separation of selenohypotaurine, selenotaurine, selenohomohypotaurine and selenohomotaurine.

By either paper or ion-exchange chromatography the two seleninic compounds selenohypotaurine and selenohomohypotaurine, and the two selenonic compounds selenotaurine and selenohomotaurine may be all separated from each other. On paper chromatography seleninic derivatives may be separated from the corresponding sulphinic compounds, while selenonic compounds show RF values similar to those of the corresponding sulphonic derivatives. These two latter types of compounds may be differentiated, however, since selenonic compounds liberate iodine from HI, while sulphonic compounds do not. Also by automated ion-exchange chromatography seleninic derivatives are well separated from the analogous sulphinic compounds, while selenonic compounds are eluted together with the corresponding sulphonic compounds.

Oxidation of selenocystamine by diamineoxidase.

Selenocystamine is oxidatively deaminated by pig kidney diamineoxidase. The first product of the reaction is the corresponding cyclized aminoaldehyde, selenocystaldimine, which then undergoes further degradation. The oxidative deamination is thus the first step of a series of cyclic reactions which give rise to extensive cleavage of selenocystamine.

Oxidation of S-e-carboxymethyl-selenocysteine by L-aminoacid oxidase and by D-aspartate oxidase.

Se-Carboxymethyl-DL-selnocysteine (CMSeC) has been prepared in a pure crystalline form from selenocysteine and monochloracetic acid. It has been shown that CMSeC is a substrate for the L-aminoacid oxidase form snake venom and for the D-aspartate oxidase from beef kidney. Oxygen consumption and ammonia production indicate that only the L or the D form of CMSeC ar acted upon respectively by one or the other of the above enzymes. No noticeable differences were shown in the oxidation rate of CMSeC and S-carboxymethylcysteine, an indication that the substitution of a selenium for a sulfur atom in the molecule does not greatly affect the substrate specificity of the two enzymes. Data have been obtained suggesting that the product of the oxidative deamination of CMSeC Is Se-carboxymethyl-selenopyruvic acid.

Synthesis of selenocystathionamine and preliminary data on its oxidation by diamineoxidase.

Details are reported for the synthesis of selenocystathionamine, the 2-aminoethyl, 3-aminopropyl selenide, NH2CH2CH2-Se-CH2CH2CH2NH2. Paper and ion exchange chromatographic behaviour of the compound has been investigated. By automated ion exchange chromatography on a sulfonated resin selenocystathionamine may be easily differentiated from cystathionamine, cystamine and selenocystamine. It has been shown that selenocystathionamine is oxidatively deaminated by pig kidney diamineoxidase. Preliminary data indicate that the first product of the reaction is a cyclized aminoaldehyde which then undergoes further degradation.