Plasma circulating fibrinogen stability and moderate beer consumption

Moderate beer consumption (MBC) is cardioprotective: it positively influences plasma lipid levels and plasma antioxidant activity in beer-consuming individuals. The connection between MBC and blood coagulation is not clearly defined. Forty-two volunteers were equally divided into experimental (EG) and control (CG) groups following coronary bypass surgery. For 30 consecutive days, only patients of the EG consumed 330 mL of beer per day (about 20 g of alcohol). A comprehensive clinical investigation of 42 patients was done. Blood samples were collected before and after the investigation for a wide range of laboratory tests. The plasma fibrinogen was denatured with 8 M urea and intrinsic fluorescence (IF), hydrophobicity and differential scanning calorimetry (DSC) were used to reveal possible qualitative changes. After 30 days of moderate beer consumption, positive changes in the plasma lipid levels, plasma anticoagulant and plasma antioxidant activities were registered in patients of the EG group. In 17 out of 21 patients of the same group, differences in plasma circulating fibrinogen's (PCF), secondary and tertiary structures were found. The stability of fibrinogen, expressed in thermodynamic parameters, has shown that the loosening of the structure takes place under ethanol and urea denaturation. Also fluorescence stability of PCF was decreased. No changes in the lipid levels, anticoagulant and antioxidant activity or changes in PCF were detected in patients of CG. In conclusion, for the first time after a short term of moderate beer consumption some qualitative changes in the plasma circulating fibrinogen were detected: differences in the emission peak response, fluorescence intensity and all thermodynamic data. Together, with the decrease in the PCF concentration it may lead to an elevation of the blood anticoagulant activity.     

The influence of alcohol-containing and alcohol-free beverages on lipid levels and lipid peroxides in serum of rats

It is an established fact that moderate consumption of alcoholic beverages leads to some positive biochemical changes in blood that are widely regarded as indicators of improved prevention of atherosclerosis. However, at present, there are different opinions regarding the biologically active compounds of alcoholic beverages that bring about these changes. This experiment was conducted on 60 male Wistar rats, which were divided into five groups, each of which contained 12 rats: four experimental groups (EG1, EG2, EG3, EG4) and one control group (CG). During 4 weeks, all groups of rats were fed basal diet (BD) supplemented with dry red wine (EG1), beer (EG2), lyophilized dry red wine (EG3), or lyophilized beer (EG4). The rats of the CG were fed BD only. The rats of EG1 and EG2 were fed BD supplemented daily with 2.0 mL of wine and 6.0 mL of beer, respectively. The rats of EG3 and EG4 were fed BD supplemented daily with lyophilized wine and lyophilized beer at a concentration corresponding to an intake of 2.0 mL of original wine and 6.0 mL of original beer, respectively. Before and after completion of the trial, a wide range of laboratory tests including lipids and lipid peroxides were performed. The results of this investigation reveal that both original and lyophilized wine and beer exercise statistically significant beneficial lipidemic and antioxidant effects by reducing total cholesterol (TC), low density lipoprotein cholesterol, triglycerides, and lipid peroxides (P < 0.05 for all) and by elevating the high density lipoprotein cholesterol:TC ratio. There were no statistically significant differences in the results between groups fed BD supplemented with original wine and beer versus groups fed BD supplemented with lyophilized wine and beer. Therefore, it can be concluded that the biologically active compound of these beverages is their dry matter containing inter alia polyphenols in relatively high concentrations.     

Biomass, protein- and carbohydrate-composition of phytoplankton in Varna Bay, Black Sea

It was shown that phytoplankton from the Varna Bay, Black Sea, has significantly more suspended carbohydrates, proteins and biomass in July than in April. The dominant species were Bacillariophyceae and Dinophyceae. Electrophoretic and fluorescent spectra have shown the main differences in molecular weight and stability of phytoplankton proteins. Phytoplankton included specific proteins distributed over a limited range of molecular weights between 14 and 72 kilodaltons (kDa). The most abundant protein constituents in phytoplankton samples collected in April were around 45–55 kDa. The seasonal variations of the environment influence the quantitative and qualitative changes in phytoplankton.     

Identification of pathogens in culture-negative infective endocarditis cases by metagenomic analysis

Background

Pathogens identification is critical for the proper diagnosis and precise treatment of infective endocarditis (IE). Although blood and valve cultures are the gold standard for IE pathogens detection, many cases are culture-negative, especially in patients who had received long-term antibiotic treatment, and precise diagnosis has therefore become a major challenge in the clinic. Metagenomic sequencing can provide both information on the pathogenic strain and the antibiotic susceptibility profile of patient samples without culturing, offering a powerful method to deal with culture-negative cases.

Methods

To assess the feasibility of a metagenomic approach to detect the causative pathogens in resected valves from IE patients, we employed both next-generation sequencing and Oxford Nanopore Technologies MinION nanopore sequencing for pathogens and antimicrobial resistance detection in seven culture-negative IE patients. Using our in-house developed bioinformatics pipeline, we analyzed the sequencing results generated from both platforms for the direct identification of pathogens from the resected valves of seven clinically culture-negative IE patients according to the modified Duke criteria.

Results

Our results showed both metagenomics methods can be applied for the causative pathogen detection in all IE samples. Moreover, we were able to simultaneously characterize respective antimicrobial resistance features.

Conclusion

Metagenomic methods for IE detection can provide clinicians with valuable information to diagnose and treat IE patients after valve replacement surgery. However, more efforts should be made to optimize protocols for sample processing, sequencing and bioinformatics analysis.

     

Some analytical assays for the determination of bioactivity of exotic fruits

Introduction – The consumption of new exotic fruits, with their high nutritional and sensory value, has significantly increased in the past few years. Among the tropical fruits durian (Durio zibethinus Murr.) is less known than mango (Mangifera indica L.) and avocado (Persea americana). It has been shown that durian, mango and avocado possessed high nutritional and bioactive properties, but these data were determined using different methods. In order to obtain reliable results we investigated samples of durian, mango and avocado of the same stage of ripeness and unified methods were used for determination of the antioxidant potential. As far as we know, no results of such comparative investigation of three tropical fruits (durian, mango and avocado) and the use of such tests for phytochemical control have been published. Objective – Lyophilised durian, mango and avocado samples harvested in 2008 in Thailand and Israel were investigated. Methodology – The contents of crude protein, fat, carbohydrate, dietary fibre, total polyphenols, flavonoids, tannins and flavanols were determined by elemental analysis and UV spectroscopy. The presence of polyphenols (flavonoids and phenolic acids) in the investigated samples was studied by Fourier transform infrared (FT‐IR) spectroscopy and three‐dimensional fluorometry. Four complementary radical scavenging assays were used for antioxidant determination: ferric reducing antioxidant power (FRAP), 2, 2‐azino‐bis (3‐ethyl‐benzothiazoline‐6‐sulfonic acid) diamonium salt (ABTS^?+), 1‐diphenyl‐2‐picrylhydrazyl method (DPPH) and cupric reducing antioxidant capacity (CUPRAC). Chemometrical processing was used for statistical comparison of the fruits. Results – All spectrometric measurements were highly correlated. The contents of total fibre, proteins and fats were significantly higher (p < 0.05) in avocado, and carbohydrates were significantly lower in avocado (p > 0.05) than in the two other fruits. The wavelength numbers of FTIR spectra for three investigated fruits were in the same range (1700–600 cm^?1) as for catechin and gallic acid, used as standards. One main peak could be easily observed at the approximate location of ex/em 275/305 nm and the other one at ex/em 350/430 nm in the methanol polyphenol extracts of investigated fruits in three‐dimensional fluorescence, in contour and cross fluorescence maps. Similarity was found between durian, mango and avocado in polyphenols (9.88 ± 1.0, 12.06 ± 1.3 and 10.69 ± 1.1, mg gallic acid equivalents/g dry weight, d.w.), and in antioxidant assays such as CUPRAC (27.46 ± 2.7, 40.45 ± 4.1 and 36.29 ± 3.7, µM Trolox equivalent (TE)/g d.w.) and FRAP (23.22 ± 2.0, 34.62 ± 3.4 and 18.47 ± 1.9, µM TE/g d.w.), respectively. The multisample median test between all possible pairs of groups is a Tukey–HSD type comparison and denotes the different groups in a case when a pair‐wise test is significant and its q statistical value is greater than the table q parameter. The multisample median test of FRAP values were chosen from the compared fruits triplets as similar or homogenous subsets durian and avocado. Conclusion – Nutritional and bioactive values of durian are comparable with these indices in mango and avocado. These fruits contain high, comparable quantities of basic nutritional and antioxidant compounds, and possess high antioxidant potentials. All fruits show a high level of correlation between the contents of phenolic compounds and the antioxidant potential. The methods used (three‐dimensional fluorescence, FTIR spectroscopy, radical scavenging assays) are suitable for bioactivity determination of these fruits. In order to receive best results, a combination of these fruits has to be included in the diet. The methods used are applicable for bioactivity determination in phytochemical analysis in general. Copyright © 2010 John Wiley & Sons, Ltd.     

Evaluation of some cereals, plants and tubers through protein composition

Wild and cultivated maize, sorghum, rice, amaranth, soybean, and cassava were screened for variability in seed storage proteins. Total seed proteins, albumin (Alb-1 and Alb-2), globulin, alcohol-soluble (A1 and A2), and glutelin (G1 and G2) fractions were investigated by means of sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). The comparison was done by the obtained protein patterns and their relative amounts. Using quantitative analysis of the protein composition and the electrophoretic patterns, the relationships between total proteins and amount of individual proteins were determined. Electrophoretic patterns of extracted proteins from investigated samples showed that the main protein subunits were concentrated between 10 and 45 kDa. Variation was found in major fractions and minor bands. Electrophoretic patterns of the protein fractions are directly related to the genetic background of the protein and can be identified and used to certify the genetic makeup of wild, cultivated, or newly derived cereals and plants.     

Raw and boiled garlic enhances plasma antioxidant activity and improves plasma lipid metabolism in cholesterol-fed rats

In the present study the effect of garlic, in a form more similar to how most people eat garlic, on lipid and antioxidant metabolism in rats was investigated. The antioxidant activity was determined by the efficacy to scavenge 2, 2′-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) derived radicals in garlic samples. The highest results were estimated in aqueous fraction in comparison with other extracts divided on the basis of polarity. Wistar male rats were randomly divided into 10 diet groups, each with seven animals. The groups were named: Control, RG (raw garlic), BG (boiled garlic for 20 min), AERG (aqueous extract of raw garlic), AEBG (aqueous extract of boiled garlic), Ch (Cholesterol), Ch/RG, Ch/BG, Ch/AERG and Ch/AEBG. All experimental diets were supplemented with 25 mg of lyophilized garlic/kg body weight obtained from raw, boiled and their aqueous extracts over a period of 30 days.Serum lipid (total cholesterol, LDL-cholesterol and triglycerides) concentrations were higher in all groups fed cholesterol (Ch); however, the increase was significant only in Ch group, without garlic supplementation. In groups of rats fed diets with cholesterol, garlic samples significantly hindered the rise of TC and LDL-C (P < 0.05). A significant increase (P < 0.05) in the plasma antioxidant activity was registered in experimental groups of rats fed cholesterol-free diets supplemented with garlic; oppositely, a significant decrease was only in group of rats given food containing cholesterol without garlic.The protein spectra has shown that during short boiling some proteins change their functional properties such as solubility and mobility, resulting in a number of protein bands in SDS-electrophoresis.

Conclusions

Raw and boiled garlic improved plasma lipid metabolism and plasma antioxidant activity in an experiment on rats. Thus, dietary hypolipidemic garlic was effective in reducing the oxidant stress, which was indicated by an increase of antioxidant activity and a decrease of lipids in the rats' blood. It was found that garlic boiled for 20 min has the same bioactivity as raw garlic in its antioxidant and protein spectra. Therefore it should be added at this time to foods. The selenium and copper content of raw garlic is not altered by boiling. The protein electrophoretic pattern of raw garlic is altered by boiling.     

Bioactivity of wine prepared from ripened and over-ripened kiwifruit

In order to evaluate the fruit maturity value of ‘Hayward’ kiwifruit (Actinidia deliciosa) as material for wine production, ripened and over-ripened fruit samples were analyzed. In addition, the effect of pectinase enzyme treatment on physicochemical characteristics of the wine during the fermentation process was also examined. The results showed that wine production from ripening kiwifruit took almost twice as long as from over-ripened fruit. The yield of wine production was increased from 63.35% to 66.19% by using riper kiwifruits and adding pectinase before amelioration. The soluble solid content (13.5%) in the wine produced from over-ripened fruits increased in comparison with the wine produced from ripened samples (10.3%). In both groups there was a decrease to 5° Brix at the end of fermentation. Difference of fruit maturity did not have a significant effect on acidy, pH, nor color of wine. The highest scores of total acceptance estimated during fermentation were for wine made from over-ripened fruits. Total phenolics content and antioxidant activities were similar in both wine groups while the mineral content, especially potassium, in wine made from over-ripened kiwifruit was higher than wine produced from ripened fruits. In conclusion, the quality characteristics of kiwifruit wine made from over-ripened fruit treated with pectinase showed higher values of wine in many aspects such as sensory value, alcohol and total phenolics content, antioxidant activity, minerals and production yield.     

Stability of Some Cactaceae Proteins Based on Fluorescence, Circular Dichroism, and Differential Scanning Calorimetry Measurements

Characterization of three cactus proteins (native and denatured) from Machaerocereus gummosus (Pitahaya agria), Lophocereu schottii (Garambullo), and Cholla opuntia (Cholla), was based on electrophoretic, fluorescence, CD (circular dichroism), DSC (differential scanning calorimetry), and FT-IR (Fourier transform infrared) measurements. The obtained results of intrinsic fluorescence, DSC, and CD were dissimilar for the three species of cactus, providing evidence of differences in secondary and tertiary structures. Cactus proteins may be situated in the following order corresponding to their relative stability: Machaerocereus gummosus (Pitahaya agria) > Cholla opuntia (Cholla) > Lophocereu schottii (Garambullo). Thermodynamic properties of proteins and their changes upon denaturation (temperature of denaturation, enthalphy, and the number of ruptured hydrogen bonds) were correlated with the secondary structure of proteins and disappearance of -helix.     

Intrinsic tryptophan fluorescence of human serum proteins and related conformational changes

The unfolding of human serum proteins (HSP) was studied by measuring the intrinsic fluorescence intensity at a wavelength of excitation corresponding to tryptophan's or typosine's fluorescence and surface hydrophobicity. The maxima emission wavelengths (_max) of human serum albumin (HSA) and human serum globulin (HSG) before beer consumption (BC) were 336.0 and 337.0 nm and after BC shifted to 335.0 and 334.0 nm, respectively. The surface hydrophobicity slightly increased after BC. In a solution of 8 M urea the _max of BSA shifted to 346.4 and that of BSG to 342.5 nm. In contrast, in the same solution but after BC the _max positions of HSA and HSG shifted to 355.9 and 357.7 nm, respectively. A decrease in fluorescence intensity, a shift in the maximum of emission, and an increase in surface hydrophobicity which reflected unfolding of proteins were observed. Here we provide evidence that the loosening of the HSP structure takes place primarily in various concentrations of urea before and after beer consumption. Differences in the fluorescence behavior of the proteins are attributed to disruption of the structure of proteins by denaturants as well as by the change in their compactability as a result of ethanol consumption.