The Influence of Geographical Origin on Poplar Propolis Composition and the Impact of Human Microbiota.

Ethanol extracts obtained from 13 poplar propolis samples originating from various European countries by traditional maceration were tested for total polyphenols, flavonoid content, and antioxidant activity. Moreover, the content of 18 polyphenolic compounds (from the group of phenolic acids and flavonoids) was determined using the HPLC method. The inhibitory effect of six selected extracts with the highest activity was assessed by well-diffusion method against five strains (Bifidobacterium spp., L. rhamnosus, L. acidophilus, E. coli, and Bacteroides spp.) of intestinal bacteria self-isolated from the faeces of obese probands with the use of selective media. It was found that the antioxidant activity of propolis varied depending on geographical origin and even among samples from the same region, which indicates that some other factors also influence propolis quality. The samples of different geographical origin varied mainly in the share of individual phenolic compounds, and it was not possible to find a characteristic marker of origin, excluding the galangin present in the Polish samples only. Assessing the inhibitory activity of propolis (in the range of 70 mg to 10 µg per mL) indicated that the concentration of 100 µg/mL was found as being safe for tested fecal bacteria (Bifidobacterium spp., L. rhamnosus, L. acidophilus, E. coli, and Bacteroides spp.). As no negative effect of low doses of propolis on the intestinal microflora was found, it can be suggested that its use in recommended doses brings only beneficial effects to the body.

Effect of Caffeine on the Inflammatory-Dependent Changes in the GnRH/LH Secretion in a Female Sheep Model.

Caffeine is one of the most widely consumed psychoactive drugs in the world. It easily crosses the blood-brain barrier, and caffeine-interacting adenosine and ryanodine receptors are distributed in various areas of the brain, including the hypothalamus and pituitary. Caffeine intake may have an impact on reproductive and immune function. Therefore, in the present study performed on the ewe model, we decided to investigate the effect of peripheral administration of caffeine (30 mg/kg) on the secretory activity of the hypothalamic-pituitary unit which regulates the reproductive function in females during both a physiological state and an immune/inflammatory challenge induced by lipopolysaccharide (LPS; 400 ng/kg) injection. It was found that caffeine stimulated (p < 0.01) the biosynthesis of gonadotropin-releasing hormone (GnRH) in the hypothalamus of ewe under both physiological and inflammatory conditions. Caffeine also increased (p < 0.05) luteinizing hormone (LH) secretion in ewes in a physiological state; however, a single administration of caffeine failed to completely release the LH secretion from the inhibitory influence of inflammation. This could result from the decreased expression of GnRHR in the pituitary and it may also be associated with the changes in the concentration of neurotransmitters in the median eminence (ME) where GnRH neuron terminals are located. Caffeine and LPS increased (p < 0.05) dopamine in the ME which may explain the inhibition of GnRH release. Caffeine treatment also increased (p < 0.01) cortisol release, and this stimulatory effect was particularly evident in sheep under immunological stress. Our studies suggest that caffeine affects the secretory activity of the hypothalamic-pituitary unit, although its effect appears to be partially dependent on the animal's immune status.

The Use of HPTLC and SDS-PAGE Methods for Coniferous Honeydew Honey Fingerprinting Compiled with Mineral Content and Antioxidant Activity.

Fir honeydew honey is a uniquely beneficial product which is often subjected to adulteration; however, pollen analysis is not useful to verify this honey type. Fourteen samples of EU protected designation of origin fir honeydew honey gathered directly from apiaries were studied. Standards of legal requirements and additional parameters, i.e., specific optical rotation, mineral content, and antioxidant activity, were tested. Five nectar honeys of different varieties were used as a comparative material. HPTLC and SDS-PAGE methods were used to fingerprint the honey types. All honeys tested fulfilled the quality requirements in terms of water content, pH, total acidity, conductivity, HMF, and diastase number. They were defined as dark amber on the Pfund scale and exhibited positive specific rotation (+2.5 to 25). Honeydew honey surpassed the tested nectar honeys in terms of mineral content and antioxidant activity as well as total polyphenolic content, except for buckwheat honey. The sugar and polyphenolic profile obtained by HPTLC allowed to distinguish honeydew from nectar honeys. The same was achieved by SDS-PAGE protein profiling. Both techniques seem to be cheap and quick tools for precisely distinguishing honeydew honey.

The Study of Chemical Profile and Antioxidant Properties of Poplar-Type Polish Propolis Considering Local Flora Diversity in Relation to Antibacterial and Anticancer Activities in Human Breast Cancer Cells.

Nine samples of ethanolic extracts of poplar-type propolis (EEP) originated from South-Eastern Poland were analyzed in terms of the diversity of the flora around the apiary. The mineral composition, antioxidant properties, polyphenolic profile (HPTLC), and main polyphenolic constituents (HPLC-DAD) were determined. Only minor differences in chemical composition and antioxidant capacity between tested EEPs were found regardless of their botanical origin. However, the biological activity of the EEPs was more diversified. The tested EEPs showed stronger antibacterial activity against Gram-negative bacteria (Escherichia coli) compared to Gram-positive bacteria (Staphylococcus aureus and Staphylococcus epidermidis). Staphylococci biofilm inhibition occurred as a result of exposure to the action of four out of nine EEPs (P1-P4). Due to the various compositions of individual EEPs, a different MCF-7 cellular response was observed according to inhibition of cells migration and proliferation. Almost every sample inhibited the migration of breast cancer cells at a low concentration (0.04 µg/mL) of propolis. Even at the lowest concentration (0.02 µg/mL), each EEP inhibited the proliferation of MCF-7 cells, however, the level of inhibition varied between samples.

Antioxidant Activity of Frozen and Freeze-Dried Drone Brood Homogenate Regarding the Stage of Larval Development.

Drone brood is a little-known and poorly studied bee product used and valued in the treatment of many diseases, including male infertility and women's menopausal disorders. The aim of this study was to evaluate the antioxidant activity of drone brood depending on the stage of larval development and the method of preservation. Aqueous and ethanolic homogenate extracts of drone brood were assayed for antioxidant activity (with the DPPH, FRAP, and ABTS methods), polyphenol, and flavonoid content. The extracts' polyphenolic profiles were compared by the HPTLC method. Drone brood has been shown to be more active in the earlier stages of development (between days 7-11), with a decline in antioxidant activity in the later period (by the 14th day). The freeze-drying process did not cause significant changes in the antioxidant activity of brood preparations converted to dry mass. Based on the higher activity of the aqueous compared to 70% ethanolic extracts, it was shown that the dominant fraction of brood consisted of hydrophilic antioxidants. The results obtained with different methods were highly correlated, excluding those from the ABTS assay. The HPTLC method showed that the polyphenol fraction of drone brood homogenate consisted mainly of phenolic acids and flavonoids. It was shown that drone brood has valuable antioxidant properties that can be compared with royal jelly.

The Effect of Adding the Leaves and Fruits of Morus alba to Rape Honey on Its Antioxidant Properties, Polyphenolic Profile, and Amylase Activity.

Two various species of mulberry (Morus sp.) were selected to enrich rape honey with dried leaves or lyophilized fruits (4% w/v). Finally, fruits and leaves of the 'Ukrainska' clone were introduced into the honey during creaming in concentrations from 1 to 4% w/v. The total phenolic content, antioxidant activity, anthocyanins content, and polyphenolic profile were tested in plant extracts and enriched honeys. Moreover, α-glucosidase, ß-galactosidase, and diastase activities were investigated in honeys. For mulberry extracts, chlorogenic acid isomers and rutin were considered main antioxidant compounds. The antioxidant activity of honey enriched with mulberry leaves increased even more than 50 times, due to introducing numerous phenolic acids and flavonoid glycosides. A significant decrease in the diastase activity in honey depending on the content of added mulberry leaves (almost 50% decrease in the case of 4% addition) was found, suggesting the inhibitory effect of honey with mulberry leaves against carbohydrate hydrolyzing enzymes.

A Recombinant Turkey Herpesvirus Expressing F and HN Genes of Avian Avulavirus-1 (AAvV-1) Genotype VI Confers Cross-Protection against Challenge with Virulent AAvV-1 Genotypes IV and VII in Chickens.

Newcastle disease (ND) is responsible for significant economic losses in the poultry industry. The disease is caused by virulent strains of Avian avulavirus 1 (AAvV-1), a species within the family Paramyxoviridae. We developed a recombinant construct based on the herpesvirus of turkeys (HVT) as a vector expressing two genes: F and HN (HVT-NDV-F-HN) derived from the AAvV-1 genotype VI ("pigeon variant" of AAvV-1). This recombinant viral vaccine candidate was used to subcutaneously immunize one group of specific pathogen-free (SPF) chickens and two groups of broiler chickens (20 one-day-old birds/group). Humoral immune response was evaluated by hemagglutination-inhibition test and enzyme-linked immunosorbent assay (ELISA). The efficacy of the immunization was assessed in two separate challenge studies performed at 6 weeks of age with the use of virulent AAvV-1 strains representing heterologous genotypes IV and VII. The developed vaccine candidate elicited complete protection in SPF chickens since none of the birds became sick or died during the 2-week observation period. In the broiler groups, 90% and 100% clinical protection were achieved after challenges with AAvV-1 of IV and VII genotypes, respectively. We found no obvious relationship between antibody levels and protection assessed in broilers in the challenge study. The developed recombinant HVT-NDV-F-HN construct containing genes from a genotype VI AAvV-1 offers promising results as a potential vaccine candidate against ND in chickens.

Inflammation and LPS-Binding Protein Enable the Stimulatory Effect of Endotoxin on Prolactin Secretion in the Ovine Anterior Pituitary: Ex Vivo Study.

Prolactin is a hormone that plays an important role in the regulation of many physiological processes including lactation, reproduction, fat metabolism, and immune response. The secretion of prolactin could be disturbed by an immune stress commonly accompanying infection. This study was designed to determine the influence of bacterial endotoxin-lipopolysaccharide (LPS)-on prolactin gene (PRL) expression and prolactin release from the ovine anterior pituitary (AP) explants collected from saline- and LPS-treated ewes in the follicular phase. The expressions of toll-like receptor 4 (TLR4) and proinflammatory cytokines interleukin- (IL-) 1ß, IL-6, and tumor necrosis factor- (TNF-) α genes were also assayed. The results of the study showed that LPS stimulates prolactin secretion and IL-6 gene expression in the AP explants, but its action on lactotrophs depends on the immunological status of animal. It was demonstrated that an important role in enhancing the effect of LPS on the pituitary in the saline-treated ewes is played by LPS-binding protein (LBP)- "adapter molecule" for LPS binding to the cell surface receptor CD14 and then to TLR4. Also, it was found that bacterial endotoxin acting on the anterior pituitary cells may enhance prolactin secretion, and this effect of LPS could be mediated by IL-6 which is known as prolactin-releasing factor. Identification of the neuroendocrine and immune interactions in the regulation of prolactin secretion could be helpful in developing newer and more effective treatments for dysfunctions connected with disorders in this hormone secretion.