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Background:Vaccinium myrtillus L. is a species belonging to the genus Vaccinium of the family Ericaceae. Bilberries have drawn attention due to the multiple benefits for the human health, including antioxidant, anti-inflammatory, anticancer, anti-neurodegenerative, and cardioprotective effects. Recently, bilberries were shown to inhibit the activity of carbohydrate-hydrolysing enzymes that can help reduce the intensity of the metabolic syndrome and prevent type 2 diabetes.

Background:Plantago major L. leaves have been used for centuries by the traditional medicine in the treatment of infectious disorders of the respiratory, urinary and digestive tracts. Researchers have reported that hot water extracts of Plantago major possess a broad-spectrum of anticancer, antioxidant and antiviral activities, as well as activities which modulate cell-mediated immunity. Their beneficial properties may be due to the significant content of polysaccharides. The polysaccharides that have been isolated from the leaves of Plantago major L. have different structures – pectic substances, galactans, arabinogalactans, glucomannans.

Recently, consumer preferences for bilberries have increased markedly. This fact is probably related to their natural constituents, such as phenolic compounds including anthocyanins and tannins, as well as the vitamins and minerals they contain. Phenolic compounds are known for their numerous beneficial effects on human health. Moreover, bilberry fruits have been shown to inhibit the activity of carbohydrate hydrolyzing enzymes, which can significantly decrease the postprandial increase in blood glucose levels. Thus, the aim of the present study is to investigate the inhibitory effect of Vaccinium myrtillus L. extracts on key enzyme α-amylase, linked to type 2 diabetes. No data have been published on the inhibitory properties of Vaccinium myrtillus L. fruits growing wild in Bulgaria against carbohydrate enzymes. Bilberry extracts were analyzed for total polyphenols, total anthocyanin content, antioxidant activity and their inhibitory properties against α-amylase. The contents of flavonols, anthocyanins and stilbenes were determined by HPLC analysis. The identified flavonols in the analyzed bilberry extracts were mainly represented by quercetin derivatives as rutinoside. The predominant anthocyanins for both aqueous and organic solvents were delphinidin-3-galactoside and malvidin-3-glucoside. The results revealed that bilberry extracts are effective inhibitors of α-amylase, with IC50 values from 20.8 to 194.8 μg GAE/mL. All the samples proved to have antioxidant activity measured by three different in vitro assays (FRAP, CUPRAC and DPPH). The inhibitory properties of V. myrtillus L. extracts may provide a new direction in the development and research of new pharmaceuticals for the suppression of postprandial hyperglycemia in diabetic patients. [ABSTRACT FROM AUTHOR]

This study examined the effect of lactulose, galactooligosaccharide, fructooligosaccharide, inulin, and β-glucan on the probiotic strain Lactobacillus delbrueckii subsp. bulgaricus L14, cultivated in an in vitro gastrointestinal system model. We analyzed the degree of hydrolysis of the studied prebiotic oligosaccharides in condition of simulated gastric fluid. The results showed that lactulose had the highest resistance, galactooligosaccharide underwent hydrolysis, and fructooligosaccharide was the most sensitive. Among the polysaccharides, fructose was released from inulin and glucose from β-glucan. Short-chain oligosaccharides and metabolites derived from studied prebiotic oligosaccharides and polysaccharides, supported the growth of probiotic strain L14, which showed the highest growth with fructooligosaccharides and β-glucan as carbohydrate sources. The profile of the activated enzymes secreted by the probiotic strain L14, indicated their inducible character. Beta-galactosidase was activated in the presence of lactulose and GalOS, inulinase was activated in the presence of inulin and fructooligosaccharides, and β-glucosidase was activated in the presence of β-glucan fragments. Analysis of the produced organic and short-chain fatty acids showed that the typical representative of the homofermentative lactobacilli Lb. delbrueckii subsp. bulgaricus L14 changes its metabolism from a homofermentative to a heterofermentative type, best expressed in the presence of lactulose, galactooligosaccharide, and β-glucan. [ABSTRACT FROM AUTHOR]

For the period between June 2020 and -March 2022, a total of 8 dairy cattle farms were surveyed in terms of the prevalence of clinical and subclinical mastitis. Four of them were located in Northern Bulgaria (Targovishte, Shumen and Dobrich districts) and another four in Southern Bulgaria (Stara Zagora, Plovdiv and Haskovo districts). In these target farms, a rapid mastitis screening test was initially performed to detect the presence of subclinical mastitis or samples with high somatic cell counts. A total of 312 milk samples were obtained from milk quarters who reacted with 3+ or 4+ to which 34 samples of inflammatory exudate from cows with clinical mastitis were added. During the microbiological investigation of the 346 samples, 272 of them (79.1%) were bacteriologically positive. Streptococcus spp., were isolated from 151 samples (55.5%). The second most common species was Staphylococcus spp., detected in 110 (40.4%) of the tested samples. In total, the Grampositive cocci finding exceeded 95% of the microbial species. The remaining 11 (4.0%) isolates belonged to another 6 taxa. These included four strains identified by prior phenotypic identification as Trueperella pyogenes, three Escherichia coli isolates, and one strain of each Pasteurella multocida, Nocardia asteroides, Klebsiella pneumoniae, and Acinetobacter spp. Staphylococci were also studied for their sensitivity to 11 chemotherapeutic agents. The highest percentage of resistance (50%) was determined to lincomycin, followed by that to tetracycline (37.3%), and beta-lactams ampicillin (24.5%), and oxacillin (13.6%). The resistance rates to cefoxitin and cephalotin were 5.5% and 0.9% respectively. Also, lower values of resistant strains were observed for the combination of trimethoprim/sulfonamides (7.3%), ciprofloxacin (1.8%), and rifampicin (0.9%). Resistance to gentamicin and amoxicillin/ clavulanic acid was not established. Minimum inhibitory concentrations were determined for the studied chemotherapeutics, with the highest MIC90 values of 128 µg/mL and 2 µg/mL for tetracycline and lincomycin, and the lowest MIC90 values of 0.001 µg/mL for rifampicin and ciprofloxacin, respectively. A MIC90 of 0.125 µg/mL was obtained for oxacillin, 1.5 µg/mL for trimethoprim/sulfonamides and 1.0 µg/mL for ampicillin, cephalotin and cefoxitin. [ABSTRACT FROM AUTHOR]

In this work, we reported production of glucansucrases with molecular weights of about 300 kDa by strains of fructophilic lactic acid bacteria (FLAB) Lactobacillus pentosus AG8, Lactobacillus kunkeei AG9, AG10 and AG11 isolated from the gut of honey bee Apis mellifera. From sucrose, these enzymes synthesize average to high-molecular-weight dextran-type glucans ranging from 8.5 × 105 to >4 × 106 Da and containing 16 to 22% α-(1→3)-linked glucose units, with 5 to 8% branching in their structures. Enzyme preparations from the strains were applied in the synthesis of glucooligosaccharides (GOSs) with degrees of polymerization (DP) from 3 to 7 using maltose/sucrose ratios (M/S) from 8 to 0.12. The yields and composition of the obtained oligosaccharide products were strongly influenced by the ratios of the acceptor and donor of glucose units in the reactions. The highest yields of GOSs (>50 g/L) were achieved at M/S = 1, and the lowest content of α-(1→6) linkages (71%) combined with increased proportion of the products with DP > 3 were determined at M/S = 0.5. GOSs preparations from these two ratios were able to maintain the growth and acids production of three probiotic strains of lactobacilli, suggesting potential application as prebiotics. [ABSTRACT FROM AUTHOR]

The aim of the present study was to evaluate the prebiotic potential of Plantago major L. leaves water-extractable polysaccharide (PWPs) and its lower molecular fractions. The structure of PWPs was investigated by high pressure anion exchange chromatography (HPAEC), size exclusion chromatography coupled with multi-angle laser light scattering detector (SEC-MALLS) and Fourier-transform infrared (FTIR) spectroscopy. The chemical composition and monosaccharide analyses showed that galacturonic acid was the main monosaccharide of PWPs followed by glucose, arabinose, galactose, rhamnose and xylose. FTIR study indicated a strong characteristic absorption peak at 1550 cm−1 corresponding to the vibration of COO− group of galacturonic acid. The PWPs was subjected to hydrolysis using commercial enzymes to obtain P. major low molecular fraction (PLM) which was successively separated by size exclusion chromatography on Biogel P2. PWPs and PLM were examined for in vitro prebiotic activity using various assays. Results gave evidence for changes in optical density of the bacteria cells and pH of the growth medium. A heterofermentative process with a lactate/acetate ratio ranged from 1:1 to 1:5 was observed. The ability of PLM to stimulate the production of certain probiotic bacteria glycohydrolases and to be fermented by Lactobacillus sp. strains was successfully proved. [ABSTRACT FROM AUTHOR]

The leaves of Plantago species have been known as a rich source of polysaccharides, polyphenols, and iridoids. In the recent years, some authors have examined the relative influence of carbohydrate compositions and polyphenols on their antioxidant and prebiotic activity. In this study enzymatic hydrolysis of Plantago major, Plantago lanceolata, and Plantago media leaves with hemicellulase and xylanase was carried out. The chemical composition of the enzyme‐derived hydrolysis products was determined. Their antioxidant activity was evaluated using 2,2‐diphenyl‐2‐picryl‐hydrazyl‐hydrate, ferric reducing antioxidant power, and cupric reducing antioxidant capacity assays. The utilization of the carbohydrate complex from the obtained hydrolysates to stimulate the production of certain glycohydrolases from Lactobacillus plantarum probiotic strains was investigated. The HPLC analysis of the hydrolysis products showed the presence of galactose and arabinose in 1:3 and 1:4 ratios. The highest concentration of total sugars was determined in P. major hydrolysates with hemicellulase (386.71 mg/g dry weight). The antioxidant activity according to the three methods was the highest in P. media hydrolysates, which correlated with their total phenolic concentration. L. plantarum S27 showed the highest values of α‐galactosidase activity at the 24‐h fermentation process when it was cultivated in a broth with the presence of P. major hydrolysates with hemicellulase and xylanase (0.129 U/mg and 0.133 U/mg, respectively). [ABSTRACT FROM AUTHOR]

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Fruits of the plant Lycium barbarum L. (Goji berry) fruit have been widely used around the world for their medicinal purposes and as a functional food for more than 4500 years. Studies indicate the effects of the fruits of Goji berry on aging, neuroprotection, fatigue, energy expenditure, glucose control in diabetics, antioxidant properties, immunomodulation, anti-tumor activity, and cytoprotection. Some authors report that antioxidant molecules found in goji fruit might offer many health protective benefits by alleviating oxidative stress when used as an additive or in a mixture with other phytochemicals in fruit or herbal products. It was found that Goji berry fruit growing in Bulgaria has a total polysaccharide content of about 4%. The content of total polyphenol compounds in the different extracts was found to be between 5.5mg and 8mg GAE/g of dry product. Polyphenols from dry Goji berry fruit were extracted and studied by HPLC analysis for the type of polyphenols present. Antioxidant activity of extracts rich in polyphenols was determined by DPPH, FRAP and CUPRAC methods. The data was compared with a medicinal product containing an extract of Goji berry, where synergistic effects were also established. Inhibitory effects of the Goji berry extracts and their corresponding polyphenols were investigated in the presence of α-glucosidase for antidiabetic activity. The 50% inhibitory concentration found for organic extract was IC50=91.7 µg GAE/g of fruits. The study demonstrated that some of the polyphenols in the investigated extracts exhibit competitive properties towards the enzyme. [ABSTRACT FROM AUTHOR]

The current survey investigates the effect of four polysaccharides isolated from fresh leek or alcohol insoluble substances (AIS) of leek on the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) from phagocytes. The ability of the polysaccharides to activate serum complement was also investigated. Despite the lack of antioxidant activity, the pectic polysaccharides significantly decreased the production of ROS by human neutrophils. Polysaccharides isolated from AIS markedly activated RAW 264.7 macrophages for RNS production in a concentration-dependent manner. The Western blot analysis revealed that this effect was due to the stimulation of the inducible nitric oxide synthase protein expression of macrophages. The polysaccharides extracted from AIS with water showed the ability to fix serum complement, especially through the alternative pathway. It was found that the polysaccharide that has the highest complement- fixing effect is characterized by the highest content of uronic acids and the highest molecular weight. [ABSTRACT FROM AUTHOR]

Water-soluble and acid-soluble pectic polysaccharides with significant immunostimulating activity were isolated from alcohol-insoluble solids (AIS) of leek. Pectic polysaccharides characterized with high anhydrouronic acid content and common pectic sugars - L-arabinose, D-galactose, D-glucose, and L-rhamnose. It was accomplished that D-galactose content was the highest, which allowed the assumption that pectin sugar side chains were from galactan type. Enzyme modification of acid-extracted pectin was realized. Different pectin-degrading enzymes were used. The first modified pectic fraction, obtained after endopolygalacturonase action and alcohol precipitation contained rhamnogalacturonan I and had high D-galactose content. The second fraction obtained from the first, after ß-galactosidase treatment characterized with low D-galactose content. Probably this was result from ß-galactosidase hydrolysis of pectin side chains. It was proved by the action of ß-galactosidase that galactan side chains were mainly composed of ß-(1?4)-linked D-galactose. It was accomplished that rhamnogalacturonan I had higher immunostimulating activity than the initial pectic polysaccharide, which confirmed the statement that pectin hairy regions were responsible for the biological activity. The second fraction had lower D-galactose content and lower activity than the first. Probably galactan side chains were responsible for the immunostimulating activity. [ABSTRACT FROM AUTHOR]

We present versatile anycast, which allows a service running on a varying collection of nodes scattered over a wide-area network to present itself to the clients as one running on a single node. Providing a single logical address enables the client-side software to preserve the traditional service access model based on single access points. At the same time, the dynamic composition of anycast groups implemented by versatile anycast enables the server-side service infrastructure to evolve and adapt to changing network conditions. We implement versatile anycast using Mobile IPv6, which decouples the logical addresses of mobile nodes from their physical location. We exploit that decoupling to implement logical service addresses that are not bound to any physical nodes, and employ standard MIPv6 mechanisms to dynamically map each such address onto individual service nodes. Our solution enables a service to transparently hand off clients among the service nodes at the network level while preserving optimal routing between the clients and the service nodes. We demonstrate that the overhead of versatile anycasting is very low. In particular, the client-perceived handoff time is shown to be a linear function of the latencies among the client and the service nodes participating in the handoff. Copyright © 2007 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Simple Summary: Pollution of the aquatic environment is a well-known problem with a long history. Monitoring water quality relies on biota in order to provide adequative assessment and management of the water bodies. Among the different biological indicators applied, aquatic macrophytes, and particularly mosses, are in direct relationship with the environment and their use as biomonitors is well documented. In the current study, we made an attempt to apply new fast, reliable and comprehensible methods for water pollution control. Three reservoirs were selected for the following reasons: (i) they were polluted with hazardous substances (heavy metals and organic material) and (ii) they are used for fish farming and irrigation and their water quality directly affects human health. Moss-bags with the selected biomonitor Fontinalis antipyretica were exposed in the reservoirs for a period of 30 days and molecular, chemical and micromorphological markers were studied. All biomarkers tested appeared to be sensitive to the pollution. This research provided a basis for further studies on selected biomarkers towards standardization. Moss-bags were applied to study the effect of contamination in three standing water bodies in Bulgaria (Kardzhali, Studen Kladenets and Zhrebchevo Reservoirs), the first two with old industrial contamination and the last polluted with short-chain chlorinated paraffins (SCCPs). Fontinalis antipyretica Hedw. collected from background (unpolluted) site was placed in cages for a period of 30 days. The present study examined whether inorganic and organic pollution detected with moss-bags resulted in corresponding differences in molecular, chemical and micromorphological markers. Suppressed large subunit of ribulose-1,5-bisphosphate carboxylase (rbcL) expression was assessed in moss-bags from two of the reservoirs, contaminated with heavy metals. There was a decrease of the total phenolic content (TPC) in the moss-bags, which provides a basis for further studies of the chemical content of aquatic mosses. Fontinalis antipyretica also showed a response through leaf micromorphological characteristics. In the all three reservoirs, an increase of the twig leaf cell number was recorded (p ≤ 0.01 for Kardzhali and p ≤ 0.001 for Studen Kladenets and Zhrebchevo reservoirs), as well as of the stem leaf cell number in Zhrebchevo Reservoir (p ≤ 0.001). On the contrary, the width of the cells decreased in the studied anthropogenically impacted reservoirs. All three studied groups of biomarkers (molecular, chemical and micromorphological) appeared to be sensitive to freshwater pollution. The results achieved indicated that rbcL gene expression, TPC, cell number and size are promising biomonitoring tools. [ABSTRACT FROM AUTHOR]

Inflammation is the initial response of the immune system to potentially harmful stimuli (e.g., injury, stress, and infections). The process involves activation of macrophages and neutrophils, which produce mediators, such as nitric oxide (NO), prostaglandin E2 (PGE2), pro-inflammatory and anti-inflammatory cytokines. The pro-inflammatory cytokines interleukin-1β (IL-1β), interleukin 6 (IL-6), and tumor necrosis factor-α (TNF-α) are considered as biomarkers of inflammation. Even though it occurs as a physiological defense mechanism, its involvement in the pathogenesis of various diseases is reported. Rheumatoid arthritis, inflammatory bowel disease, Alzheimer's disease, and cardiovascular diseases are only a part of the diseases, in which pathogenesis the chronic inflammation is involved. Fucoidans are complex polysaccharides from brown seaweeds and some marine invertebrates, composed mainly of l-fucose and sulfate ester groups and minor amounts of neutral monosaccharides and uronic acids. Algae-derived fucoidans are studied intensively during the last years regarding their multiple biological activities and possible therapeutic potential. However, the source, species, molecular weight, composition, and structure of the polysaccharides, as well as the route of administration of fucoidans, could be crucial for their effects. Fucoidan is reported to act on different stages of the inflammatory process: (i) blocking of lymphocyte adhesion and invasion, (ii) inhibition of multiple enzymes, and (iii) induction of apoptosis. In this review, we focused on the immunemodulating and anti-inflammatory effects of fucoidans derived from macroalgae and the models used for their evaluation. Additional insights on the molecular structure of the compound are included. [ABSTRACT FROM AUTHOR]

The aim of this study was to evaluate the effects of fucoidan isolated from C. crinita on histamine-induced paw inflammation in rats, and on the serum levels of TNF-α, IL-1β, IL-6, and IL-10 in rats during systemic inflammation response. The levels of TNF-α in a model of acute peritonitis in rats were also investigated. The isolated crude fucoidan was identified as a sulfated xylogalactofucan with high, medium, and low molecular weight fractions and a content of fucose of 39.74%, xylose of 20.75%, and galactose of 15.51%. Fucoidan from C. crinita showed better anti-inflammatory effects in the rat paw edema model, and this effect was present during all stages of the experiment. When compared to controls, a commercial fucoidan from F. vesiculosus, the results also displayed anti-inflammatory activity on the 60th, 90th, and 120th minute of the experiment. A significant decrease in serum levels of IL-1β in rats treated with both doses of C. crinita fucoidan was observed in comparison to controls, whereas TNF-α concentrations were reduced only in the group treated with fucoidan from C. crinita at the dose of 25 mg/kg bw. In the model of carrageenan-induced peritonitis, we observed a tendency of decrease in the levels of the pro-inflammatory cytokine TNF-α in peritoneal fluid after a single dose of C. crinita fucoidan, but this did not reach the statistical significance margin. Single doses of C. crinita fucoidan did not alter serum levels of the anti-inflammatory cytokine IL-10 in animals with lipopolysaccharide-induced systemic inflammation.

Moss-bags were applied to study the effect of contamination in three standing water bodies in Bulgaria (Kardzhali, Studen Kladenets and Zhrebchevo Reservoirs), the first two with old industrial contamination and the last polluted with short-chain chlorinated paraffins (SCCPs). Fontinalis antipyretica Hedw. collected from background (unpolluted) site was placed in cages for a period of 30 days. The present study examined whether inorganic and organic pollution detected with moss-bags resulted in corresponding differences in molecular, chemical and micromorphological markers. Suppressed large subunit of ribulose-1,5-bisphosphate carboxylase (rbcL) expression was assessed in moss-bags from two of the reservoirs, contaminated with heavy metals. There was a decrease of the total phenolic content (TPC) in the moss-bags, which provides a basis for further studies of the chemical content of aquatic mosses. Fontinalis antipyretica also showed a response through leaf micromorphological characteristics. In the all three reservoirs, an increase of the twig leaf cell number was recorded ( p ≤ 0.01 for Kardzhali and p ≤ 0.001 for Studen Kladenets and Zhrebchevo reservoirs), as well as of the stem leaf cell number in Zhrebchevo Reservoir ( p ≤ 0.001). On the contrary, the width of the cells decreased in the studied anthropogenically impacted reservoirs. All three studied groups of biomarkers (molecular, chemical and micromorphological) appeared to be sensitive to freshwater pollution. The results achieved indicated that rbcL gene expression, TPC, cell number and size are promising biomonitoring tools.

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