Metabolic Profile of Leaves and Pulp of Passiflora caerulea L. (Bulgaria) and Their Biological Activities.

At present, there are no data in the scientific literature on studies aimed at characterizing Passiflora caerulea L. growing in Bulgaria. The present study aimed to investigate the metabolic profile and elemental composition of the leaves and pulp of this Passiflora, as well as to evaluate the antioxidant, antimicrobial and anti-inflammatory activities of its leaf and pulp extracts. The results showed that the pulp predominantly contained the essential amino acid histidine (7.81 mg g-1), while it was absent in the leaves, with the highest concentration being tryptophan (8.30 mg g-1). Of the fatty acids, palmitoleic acid predominated both in the pulp and in the leaves. A major sterol component was ß-sitosterol. Fructose (7.50%) was the predominant sugar in the pulp, while for the leaves, it was glucose-1.51%. Seven elements were identified: sodium, potassium, iron, magnesium, manganese, copper and zinc. The highest concentrations of K and Mg were in the pulp (23,946 mg kg-1 and 1890 mg kg-1) and leaves (36,179 mg kg-1 and 5064 mg kg-1). According to the DPPH, FRAP and CUPRAC methods, the highest values for antioxidant activity were found in 70% ethanolic extracts of the leaves, while for the ABTS method, the highest value was found in 50% ethanolic extracts. In the pulp, for all four methods, the highest values were determined at 50% ethanolic extracts. Regarding the antibacterial activity, the 50% ethanolic leaf extracts were more effective against the Gram-positive bacteria. At the same time, the 70% ethanolic leaf extract was more effective against Gram-negative bacteria such as Salmonella enteritidis ATCC 13076. The leaf extracts exhibited higher anti-inflammatory activity than the extracts prepared from the pulp. The obtained results revealed that P. caerulea is a plant that can be successfully applied as an active ingredient in various nutritional supplements or cosmetic products.

Physicochemical, Antioxidant, and Antimicrobial Properties of Three Medicinal Plants from the Western Part of the Rhodope Mountains, Bulgaria.

The present study examined the physicochemical, antioxidant, and antimicrobial properties of three medicinal plants: thyme (Thymus callieri Borbás ex Velen), cotton thistle (Onopordum acanthium L.), and hawthorn fruit (Crataegus monogyna Jacq.) from the Western Rhodope Mountains, Bulgaria. The first stage determined the physicochemical characteristics (moisture, ash, carbohydrates, proteins, and vitamin C) of the three herbs. The second stage investigated four types of extracts (aqueous, oil, methanolic, and ethanolic) of each herb and evaluated their total phenolic content, the presence of phenolic compounds (flavonoids and phenolic acids), their antioxidant activity, and antimicrobial properties. Thyme was characterised by the highest ash, protein, and vitamin C content (6.62%, 11.30%, and 571 mg/100 g, respectively). Hawthorn fruit showed the highest moisture and carbohydrate content (8.50% and 4.20%, respectively). The 70% ethanolic extracts of the three herbs exhibited the highest levels of phenolic compounds and, consequently, pronounced antioxidant activity, compared to the other three types of extracts. The aqueous, oil, methanolic, and ethanolic thyme extracts demonstrated the highest total phenolic content-TPC (27.20 mg GAE/g, 8.20 mg GAE/g, 31.70 mg GAE/g, and 310.00 mg GAE/g, respectively), compared to the extracts of the other two plants. These results were consistent with the highest antioxidant activity of the thyme extracts determined using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, the oxygen radical absorbance capacity (ORAC) assay, and the hydroxyl radical averting capacity (HORAC) assay (except for the oil extract examined using the DPPH method). The results from the high-performance liquid chromatography (HPLC) analysis revealed that the flavonoid quercetin-3-ß-glucoside had the highest concentration in thyme (374.5 mg/100 g), while myricetin dominated in the cotton thistle (152.3 mg/100 g). The phenolic acid content analysis showed prevalence of rosmaric acid in the thyme (995 mg/100 g), whereas chlorogenic acid was detected in the highest concentration in the cotton thistle and hawthorn fruit (324 mg/100 g and 27.7 mg/100 g, respectively). The aqueous, methanolic, and ethanolic extracts showed moderate to high antibacterial potential but limited antifungal activity. None of the oil extracts inhibited the test microorganisms used in the study.

Novel Anthranilic Acid Hybrids-An Alternative Weapon against Inflammatory Diseases.

Anti-inflammatory drugs are used to relieve pain, fever, and inflammation while protecting the cardiovascular system. However, the side effects of currently available medications have limited their usage. Due to these adverse effects, there is a significant need for new drugs. The current trend of research has shifted towards the synthesis of novel anthranilic acid hybrids as anti-inflammatory agents. Phenyl- or benzyl-substituted hybrids exerted very good anti-inflammatory effects in preventing albumin denaturation. To confirm their anti-inflammatory effects, additional ex vivo tests were conducted. These immunohistochemical studies explicated the same compounds with better anti-inflammatory potential. To determine the binding affinity and interaction mode, as well as to explain the anti-inflammatory activities, the molecular docking simulation of the compounds was investigated against human serum albumin. The biological evaluation of the compounds was completed, assessing their antimicrobial activity and spasmolytic effect. Based on the experimental data, we can conclude that a collection of novel hybrids was successfully synthesized, and they can be considered anti-inflammatory drug candidates-alternatives to current therapeutics.

Physicochemical, microbiological, and sensory characteristics of probiotic Bulgarian yoghurts obtained by ultrafiltration of goat's milk.

The purpose of this study was to investigate the microbiological (number of viable lactic acid bacteria and bifidobacteria), physicochemical (dry matter, protein and fat contents, titratable acidity, and pH), and sensory characteristics (appearance of coagulum, taste and aroma, structure at cutting, color, and consistency at shattering) of probiotic Bulgarian yoghurts obtained by ultrafiltration of goat's milk. These yoghurts were obtained using volume reduction ratios of 2 and 3 with the probiotic starters MZ2f, MZ2f + Bifidobacterium bifidum BB - 87, and MZ2f + Lactobacillus acidophilus LAB - 8. The increase in the level of the concentration by ultrafiltration led to an increase in the dry matter, protein and fat contents of the yoghurts, in the number of lactic acid bacteria and titratable acidity, and to a decrease in the pH. The twofold concentration by ultrafiltration resulted in a higher number of lactic acid bacteria in comparison with yoghurts made without ultrafiltration, and with better sensory characteristics compared to yoghurts without and with threefold ultrafiltration. The higher number of viable cells and better sensory characteristics were obtained for yoghurts with MZ2f + Bifidobacterium bifidum BB - 87 and MZ2f + Lactobacillus acidophilus LAB - 8 in comparison with MZ2f alone.

Isolation, Characterization and Amino Acid Composition of a Bacteriocin Produced by Bacillus methylotrophicus Strain BM47.

Members of the bacterial genus Bacillus are known as producers of a broad spectrum of antibiotic compounds of proteinaceous nature that possess inhibitory activity against different saprophytic and pathogenic microorganisms. In the current research, a peptide synthesized by Bacillus methylotrophicus strain BM47, previously isolated from a natural thermal spring in Bulgaria, was identified and characterized as a bacteriocin. In vitro antimicrobial screening of the crude bacteriocin substance of B. methylotrophicus BM47 showed activity against the plant pathogenic fungi Fusarium moniliforme, Aspergillus awamori, Penicillium sp., Aspergillus niger and Gram-negative bacterium Pseudomonas aeruginosa. The antimicrobial activity of the crude bacteriocin substance was partially inhibited by the enzymes trypsin, Alcalase®, Savinase®, proteinase K, papain and Esperase®, while catalase was not effective. The crude bacteriocin substance was relatively pH resistant, but sensitive to the action of heat and most organic solvents and detergents tested. To obtain the active protein fractions, crude bacteriocin substance was purified by fast protein liquid chromatography (FPLC) using a strong anion exchange column. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis demonstrated that the purified bacteriocin had molecular mass of 19 578 Da. The amino acid analysis performed by high-performance liquid chromatography (HPLC) revealed that the isolated bacteriocin consisted of 17 types of amino acids, with the highest mol fraction expressed as percent of serine (29.3), valine (10.3), alanine (9.8) and tyrosine (7.1).