Chemical composition and antibacterial activity of bee venom against multi-drug resistant pathogens.

Bee venom with an antimicrobial effect is a powerful natural product. One of the most important areas where new antimicrobials are needed is in the prevention and control of multi-drug resistant pathogens. Today, antibacterial products used to treat multi-drug resistant pathogen infections in hospitals and healthcare facilities are insufficient to prevent colonisation and spread, and new products are needed. The aim of the study is to investigate the antibacterial effect of the bee venom (BV), a natural substance, on the species of Methicillin resistant Staphylococcus aureus, Vancomycin resistant Enterococcus faecalis, Carbapenem resistant Escherichia coli, Carbapenem resistant Klebsiella pneumoniae and Carbapenem resistant Acinetobacter baumannii. As a result of this study, it was found that MIC90 and MBC90 values ranged from 6.25 µg/mL - 12.5 µg/mL and numbers of bacteria decreased by 4-6 logs within 1-24 h for multi-drug resistant pathogens. In particular, Vancomycin resistant Enterococcus faecalis isolate decreased 6 log cfu/mL at 50 µg/mL and 100 µg/mL concentrations in the first hour. The effective bacterial inhibition rate of bee venom suggests that it could be a potential antibacterial agent for multi-drug resistant pathogens.Contribution: The treatment options of antibiotic-resistant pathogens are a major problem in both veterinary and human medicine fields. We have detected a high antibacterial effect against these agents in this bee venom study, which is a natural product. Apitherapy is a fashionable treatment method all over the world and is used in many areas of health. Bee venom is also a product that can be used as a drug or disinfectant raw material and can fill the natural product gap that can be used against resistant bacteria.

NMR metabolomics analysis of Escherichia coli cells treated with Turkish propolis water extract reveals nucleic acid metabolism as the major target.

AIMS: Propolis is a resinous bee product containing several hundred biologically active compounds. Although the antibacterial activity of propolis has been demonstrated in many in vitro studies, less is known about its mode of action. In this study, we aimed to shed some light on the antibacterial mechanism of action of propolis against Escherichia coli BW25113 using a nuclear magnetic resonance (NMR) based metabolomics approach. METHODS: E. coli BW25113 cells were subjected to different sub-lethal concentrations (0, 2, 4, and 6 mg/mL) of Turkish propolis water extract (PWE). The 500-MHz 1H NMR spectroscopy was then employed to ascertain the metabolic profiles of E. coli extracts. RESULTS: A total of 52 metabolites were identified from the NMR spectra, belonging to 17 main classes, such as amino acids and peptides, purines, and fatty acids. Twelve out of these 52 metabolites displayed remarkable changes at all tested PWE concentrations when compared to control conditions (P < .05). Levels of 28 more metabolites were significantly altered in at least one of the three PWE treatments. The results of partial least squares discriminant analysis showed that there was a clear separation between control and propolis-treated cells and that putrescine, adenine, adenosine, guanosine, glucose, N6-acetyllysine, and acetamide had the highest effect on group differentiation. Finally, quantitative pathway analysis revealed that purine metabolism was significantly affected by PWE treatments. CONCLUSIONS: Our results suggest that PWE inhibits the growth of E. coli BW25113 by affecting nucleic acid metabolism to a great extent. To the best of our knowledge, this is the first study to evaluate the global metabolic response of a bacterium to propolis.

Determination of the Effects of Bee Venom on Triple Negative Breast Cancer Cells in Vitro.

Honeybees provide multiple products such as bee venom (BV) which are used for various nutritional and medicinal purposes. BV has received great attention due to its wide range of bioactive components with potential anti-cancer effects on different cancers. Triple negative breast cancer (TNBC) is defined as an aggressive type of breast cancer and new therapeutic targets are required for its treatment. In the current literature information is varied about the composition and quantity of BV bioactive compounds as well as the origin of BV and its significance. In this context, the cytotoxic and apoptotic effects of BV with a higher rate of mellitin from Apis mellifera anatoliaca (Mugla ecotype) on MDA-MB-231 cells was evaluated, in vitro. The cytotoxic, apoptotic and morphological effects of BV were determined by WST-1, Annexin V, cell cycle analysis and Acridine Orange staining. The results showed that BV caused apoptotic cell death in TNBC cells at a lower dose (0.47 µg/mL, p<0.01). This study suggests that BV could be developed as a potential therapeutic agent for cancer treatment. However, the mechanism of BV-induced apoptosis death should be clarified at the molecular level.

Apis mellifera anatoliaca Venom Exerted Anti-Inflammatory Activity on LPS-Stimulated Mammalian Macrophages by Reducing the Production of the Inflammatory Cytokines.

Extraction and characterization of natural products provide the opportunity to expand our arsenal of drug candidates against a wide range of diseases including cancer and inflammatory disorders. Previous studies have shown bee venom to have immense potential as an anti-inflammatory drug candidate. In this study, we focused on the venom of Apis mellifera anatoliaca and characterized its content by HPLC. An in vitro inflammation model based on lipopolysaccharide (LPS)-stimulated mammalian macrophages was utilized to examine the venom's anti-inflammatory potential. Additionally, its antiproliferative activity was evaluated in vitro against a human glioblastoma cell line. Based on the TNF, IL6, GMCSF, and IL12p40 pro-inflammatory cytokine production level in LPS-induced macrophages, venom-treated groups showed substantial decrease in the inflammatory action compared to untreated LPS-stimulated macrophages. When the cells were analyzed for viability, the venom did not have any cytotoxic effect on the macrophages at the concentration ranges that were utilized. Moreover, IC50 value of the venom was above 60 µg/mL on glioblastoma cancer cell line. These results suggest that the Apis mellifera anatoliaca venom does not have anticancer drug candidate potential, whereas it can efficiently be used against inflammatory and autoimmune disorders. To our knowledge, this is the first study to specifically examine the effect of anti-inflammatory activity of Apis mellifera anatoliaca venom on macrophages.

The positive effect of black seed (Nigella sativa L.) essential oil on thyroid hormones in rats with hypothyroidism and hyperthyroidism.

In our study, the effect of essential oil obtained from Nigella sativa L. (NSE) on thyroid hormones and antioxidant balance in hypothyroidism (HT) and hyperthyroidism (HP) models induced by propylthiouracil(PTU) and L-thyroxine(LT4 ), respectively, in rats were investigated for 4 weeks. NSE was administered by gastric gavage at a dose of 200 mg/kg body weight. In this study, 48 male Wistar albino rats with an average weight of 180-290 g and age 5-6 months were divided into eight groups, as follows: groups with HT, (1) control, (2) HT, (3) NSE, and (4) HT + NSE; groups with HP, (1) control, (2) HP, (3), and NSE (4) HP + NSE. As a result, we found that NSE administration increased total triiodothyronine (TT3 ) and decreased nitric oxide in HT + NSE. Besides, it decreased TT3 in HP + NSE and increased total antioxidant capacity. Our findings suggest that NSE may have beneficial effects on thyroid gland abnormalities owing to its antioxidant properties. PRACTICAL APPLICATIONS: Essential oils derived from Nigella sativa L. seed contain many bioactive substances such as thymoquinone and cymene. This paper emphasizes the effect of NSE on thyroid hormone abnormalities and negative oxidative state that occurs in HT and HP models. The present study provides evidence of a positive effect of NSE particularly on TT3 levels in the HT and HP models. It can therefore be assumed that NSE could be used as a supportive natural alternative source to improve thyroid hormone levels and relieve increased oxidative stress.

Assessment of Bioactive Compounds and Antioxidant Activity of Turkey Tail Medicinal Mushroom Trametes versicolor (Agaricomycetes).

Trametes versicolor is important for its medicinal rather than nutritional value. It has a variety of pharmacological activities. The purpose of this study was to elucidate the bioactive properties of the wild medicinal mushroom T. versicolor (L.) Lloyd. Samples were analyzed for antioxidant potential and the chemical composition of the major bioactive chemical components. Chromatographic procedures were used to analyze phenolic compounds, free amino acids, vitamins, and fatty acids. T. versicolor was analyzed for moisture (87.21 ± 1.08 g/100 g fw), protein (11.07 ± 0.85 g/100 g dw), and fat (1.35 ± 0.09 g/100 g dw) content. Phenolic compounds in T. versicolor were found as p-hydroxy benzoic (113.16 ± 0.22 µg/g dw), protocatechuic (10.07 ± 0.54 µg/g dw), vanillic (5.21 ± 0.10 µg/g dw), and homogentisic acids (1.24 ± 0.15 µg/g dw). In the studied mushroom, essential and nonessential amino acids were determined as leucine (72.41 ± 0.16 mg/100 g dw), isoleucine (60.07 ± 0.10 mg/100 g dw), methionine (53.51 ± 0.09 mg/100 g dw), tyrosine (33.37 ± 0.27 mg/100 g dw), glutamine (15.48 ± 0.19 mg/100 g dw), and asparagine (10.90 ± 0.13 mg/100 g dw). Vitamin content of T. versicolor was found as nicotinic acid (26.52 ± 0.10 mg/100 g dw) and nicotinamide (12.18 ± 0.05 mg/100 g dw). Fatty acids in T. versicolor were linoleic acid (18:2n6c), oleic acid (18:1n9c), palmitic acid (C16:0), stearic acid (C18:0), and linolenic acid (18:3n3). In conclusion, the results of the present study support the potential use of T. versicolor as a promising source of bioactive products for pharmaceutical and also food industries.

Serum indoleamine 2,3 dioxygenase and tryptophan and kynurenine ratio using the UPLC-MS/MS method, in patients undergoing peritoneal dialysis, hemodialysis, and kidney transplantation.

BACKGROUND: The level and activity of indoleamine 2,3-dioxygenase (IDO) and the concentrations of L-tryptophan and its metabolite L-kynurenine were determined in association with various renal diseases. However, there have been no data regarding these parameters in patients on peritoneal dialysis compared to those undergoing hemodialysis or kidney transplantation. METHODS: This study investigated the level and activity of IDO and determined oxidative balance by calculating the total oxidant status (TOS), total antioxidant status (TAS), and oxidative stress index (OSI). We enrolled 60 kidney disease patients, including 20 on peritoneal dialysis (PD group), 19 on hemodialysis (HD group), and 21 with kidney transplantation (KT group), as well as 21 control group. RESULTS: IDO levels were increased in the PD, HD, and KT groups compared to the control group. The concentration of kynurenine was significantly increased in the PD group compared to the other groups (p < 0.01). The kynurenine/tryptophan ratio was increased in the PD group compared to the other groups (all p < 0.01). TAS levels in the PD and HD groups were significantly decreased compared to the control group (both p < 0.05). TAS levels in the PD group were significantly decreased compared to the KT group. TOS levels in the PD group were higher than in the HD and KT groups. CONCLUSION: The results showed that IDO levels were increased in peritoneal dialysis and hemodialysis patients and in renal transplant recipients, while oxidative stress was found to be related to IDO activity and was most increased in the patients on peritoneal dialysis.

Bioactive Compounds, Chemical Composition, and Medicinal Value of the Giant Puffball, Calvatia gigantea (Higher Basidiomycetes), from Turkey.

In this study, the compositions of Calvatia gigantea were first analyzed in order to elucidate its chemical basis for development as a health-enhancing food or medicine. This study investigates the chemical composition (nutritional value; phenolic, sugar and fatty acid content; aroma compounds) and antioxidant properties (radical scavenging activity, reducing power, and inhibition of lipid peroxidation) of C. gigantea. The results showed that C. gigantea contains phenolic compounds and sugars and is rich in polyunsaturated fatty acids (67.93%), proteins (34.37%), and carbohydrates (51.97%). The most abundant compounds were gentisic acid (23.26 µg/g; as a phenolic compound), trehalose (9.78 g/100g; as a sugar), and hexanal (34.71%; as an aroma compound). These findings suggest that C. gigantea might be a promising source of medicine and has the potential to be a health food and food supplementary product.

Chemical constituents: water-soluble vitamins, free amino acids and sugar profile from Ganoderma adspersum.

Ganoderma adspersum presents a rigid fruiting body owing to chitin content and having a small quantity of water or moisture. The utility of bioactive constituent of the mushroom can only be available by extraction for human usage. In this study, carbohydrate, water-soluble vitamin compositions and amino acid contents were determined in G. adspersum mushroom. The composition in individual sugars was determined by HPLC-RID, mannitol (13.04 g/100 g) and trehalose (10.27 g/100 g) being the most abundant sugars. The examination of water-soluble vitamins and free amino acid composition was determined by UPLC-ESI-MS/MS. Essential amino acid constituted 67.79% of total amino acid, which is well worth the attention with regard to researchers and consumers. In addition, G. adspersum, which is also significantly rich in B group vitamins and vitamin C, can provide a wide range of notable applications in the pharmaceutics, cosmetics, food and dietary supplement industries. G. adspersum revealed its value for pharmacy and nutrition fields.

Free amino acid profiling in the giant puffball mushroom (Calvatia gigantea) using UPLC-MS/MS.

Wild edible and medicinal mushroom, Calvatia gigantea, was quantitatively analyzed for the determination of its free amino acids using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The concentrations of total free amino acids, essential and non-essential amino acids were 199.65 mg/100 g, 113.69 mg/100 g, and 85.96 mg/100 g in C. gigantea, respectively. This study showed that C. gigantea, so called a giant puffball mushroom, has free amino acids content. The essential amino acids: tryptophan, isoleucine, valine, phenylalanine, leucine, threonine, lysine, histidine, methionine, and the non-essential amino acids: tyrosine, 4-hyrdroxy proline, arginine, proline, glycine, serine, alanine, glutamine, glutamic acid, aspargine, aspartic acid were detected.

Chemical composition, antimicrobial and antioxidant activities of Centaurea ensiformis Hub.-Mor. (Asteraceae), a species endemic to Mugla (Turkey).

The n-hexane, chloroform, ethyl acetate and ethyl alcohol extracts of the aerial parts of Centaurea ensiformis, endemic to Turkey, have been assessed for antimicrobial and antioxidant activities. The chemical composition of the hexane extract of this plant was determined. The antimicrobial activities on microorganisms, including multiple antibiotic resistant bacteria, were evaluated using the disc diffusion method. The antioxidant activities were determined by using four complementary in vitro assays: inhibition of 2,2-diphenyl-1-picrylhydrazyl radical, total antioxidant activity, and the total amount of phenolic and flavonoid compounds. The structure of the active extract was elucidated by: thin layer chromatography, column chromatography, gas chromatography or gas chromatography-mass spectrometry. The various extracts of C. ensiformis inhibited the growth of tested bacteria. All the C. ensiformis extracts had no effect on the yeasts. The total antioxidant activity increased with the increasing amount of the extracts (20, 40, 80 and 160 microg), which contained linoleic acid emulsion. The major compounds of the hexane extract of the plant were caryophyllene oxide (28.72%), spathulenol (17.81%), eudesmol (13.03%) and beta-bourbonene (8.51%).