The process of pollen transformation into bee bread: changes in bioactivity, bioaccessibility, and microbial dynamics.

Bee pollen and bee bread go hand in hand with health-promoting functional food consumption. Although many studies report high bioactivities of those products, the biotransformation of pollen into bee bread has not been fully understood. Limited findings are available about polyphenol bioaccessibility and microbiological interactions during the fermentation process. This study evaluated the microbial flora, antioxidant properties, and polyphenol and soluble protein bioaccessibility of pollen and bee bread harvested from the same apiary over a certain timeline. Total phenolic content, antioxidant activity and soluble protein content were reported using an in vitro digestion model involving post-gastric, serum-available, and colon-available fractions. The results obtained with the in vitro digestion model refer to the effect of the harvesting period on greater bioaccessibility of polyphenols in bee bread than in pollen at the same apiary. Lactic acid bacteria and yeast found in the samples were mostly identified as Lactobacillus kunkeei, Leuconostoc pseudomesenteroides, and Candida magnoliae using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS). The discrimination between the pollen and bee bread samples collected in the same apiary and at different harvesting periods was also revealed by Principal Component Analysis (PCA). A harvesting time-based approach was applied to the biotransformation process of pollen and bee bread, and insights into microbial dynamics and bioaccessibility were revealed for the first time under the same beehive conditions.

Targeting CoV-2 spike RBD and ACE-2 interaction with flavonoids of Anatolian propolis by in silico and in vitro studies in terms of possible COVID-19 therapeutics.

Propolis is a multi-functional bee product rich in polyphenols. In this study, the inhibitory effect of Anatolian propolis against SARS-coronavirus-2 (SARS-CoV-2) was investigated in vitro and in silico. Raw and commercial propolis samples were used, and both samples were found to be rich in caffeic acid, p-coumaric acid, ferulic acid, t-cinnamic acid, hesperetin, chrysin, pinocembrin, and caffeic acid phenethyl ester (CAPE) at HPLC-UV analysis. Ethanolic propolis extracts (EPE) were used in the ELISA screening test against the spike S1 protein (SARS-CoV-2): ACE-2 interaction for in vitro study. The binding energy values of these polyphenols to the SARS-CoV-2 spike and ACE-2 protein were calculated separately with a molecular docking study using the AutoDock 4.2.6 program. In addition, the pharmacokinetics and drug-likeness properties of these eight polyphenols were calculated according to the SwissADME tool. The binding energy value of pinocembrin was highest in both receptors, followed by chrysin, CAPE, and hesperetin. Based on the in silico modeling and ADME (absorption, distribution, metabolism, and excretion) behaviors of the eight polyphenols, the compounds exhibited the potential ability to act effectively as novel drugs. The findings of both studies showed that propolis has a high inhibitory potential against the Covid-19 virus. However, further studies are now needed.

Investigation of potential inhibitor properties of ethanolic propolis extracts against ACE-II receptors for COVID-19 treatment by molecular docking study.

The angiotensin-converting enzyme (ACE)-related carboxypeptidase, ACE-II, is a type I integral membrane protein of 805 amino acids that contains 1 HEXXH-E zinc binding consensus sequence. ACE-II has been implicated in the regulation of heart function and also as a functional receptor for the coronavirus that causes the severe acute respiratory syndrome (SARS). In this study, the potential of some flavonoids presents in propolis to bind to ACE-II receptors was calculated with in silico. Binding constants of ten flavonoids, caffeic acid, caffeic acid phenethyl ester, chrysin, galangin, myricetin, rutin, hesperetin, pinocembrin, luteolin and quercetin were measured using the AutoDock 4.2 molecular docking program. And also, these binding constants were compared to reference ligand of MLN-4760. The results are shown that rutin has the best inhibition potentials among the studied molecules with high binding energy - 8.04 kcal/mol, and it is followed by myricetin, quercetin, caffeic acid phenethyl ester and hesperetin. However, the reference molecule has binding energy of - 7.24 kcal/mol. In conclusion, the high potential of flavonoids in ethanolic propolis extracts to bind to ACE-II receptors indicates that this natural bee product has high potential for COVID-19 treatment, but this needs to be supported by experimental studies.

Glucose-mediated protein glycation: Contribution of methanolic extract of Ceratonia siliqua L. in protection and in vitro potential inhibition of acetylcholinesterase.

Chronic hyperglycemia presents the major etiology of diabetes mellitus and related complications mainly Alzheimer's disease, via the protein glycation and toxic products generated. In the current study, we investigated the eventual protective effect of the methanolic extract of Ceratonia siliqua L. (CsME) against glucose-mediated glycation in serum bovine albumin. The multi-stage glycation markers, namely fructosamines and advanced glycation end products (AGEs) levels were monitored along with measurement of thiol groups; moreover, the in vitro acetylcholinesterase (AChE) inhibition potential was carried out. HPLC was also assessed. Rutin was the main phenolic compound found in CsME. CsME showed a good capacity to inhibit AGEs, fructosamines and protected thiol groups against glycation. CsME exhibited a great AChE inhibition activity. In the present study, CsME prevented glucose-induced protein glycation, it also exhibited a good inhibition of AChE, suggesting its DM complications such as memory troubles related to AD. PRACTICAL APPLICATIONS: Neurodegenerative disorders ranging from memory troubles to Alzheimer's disease present the most diabetes mellitus complications and mainly attributed to protein glycation process. Currently, there is a strong trend to search for efficient natural sources of glycation and acetylcholinesterase inhibitors to replace the synthetic ones, whose secondary effects were shown. The present article tries to justify scientifically the wide use of Ceratonia siliqua L. in Moroccan folk medicine, demonstrating that the methanolic extract of leaves from this species presents a promising source of new natural compounds inhibiting acetylcholinesterase and acting in vitro against glycation generated compounds. Furthermore, for the first time, Rutin was the main phenolic compound found in this extract, these encouraging results should be coupled with further studies to integrate it in pharmaceutical formulations. As such, this paper should be of interest to a broad readership, including those interested in Biochemistry, Phytochemistry, pharmacology, and neurosciences.

Experimental arid land afforestation in Central Anatolia, Turkey.

The afforestation of arid lands faces many challenges, and perhaps the most important key for success is choosing one or more species that are adapted well for local environmental conditions. We explored species that would be suitable for the steppe region of Central Anatolia. Intensive site preparation included ripping the subsoil (to 80 cm) and plowing the upper soil before planting seedlings of Elaeagnus angustifolia, Robinia pseudoacacia, Fraxinus angustifolia, and Pinus nigra were used as tree species. We also tested the success of several shrub species: Amygdalus orientalis, Calligonum polygonoides, and Spartium junceum. After five growing seasons, E. angustifolia showed the highest survival, with 80% of planted seedlings remaining. For the shrubs, A. orientalis was the most successful species with a 95% survival rate. Broad-leaved trees grew a cumulative average of 34 cm in height in 5 years, whereas P. nigra seedings grew only 9 cm. The greatest height growth occurred in the shrubs, with A. orientalis gaining 40 cm in height in 5 years. Overall, E. angustifolia and A. orientalis appeared best suited for afforestation in these areas. R. pseodoacacia and F. angustifolia may also be used as alternative species.

Apitherapy products enhance the recovery of CCL4-induced hepatic damages in rats.

BACKGROUND/AIM: Our objective was to identify the antioxidant properties of honeybee products from Turkey, chestnut honey, pollen, propolis, and royal jelly, and their hepatoprotective activity against CCl4-induced hepatic damage in rats. MATERIALS AND METHODS: Animals were fed with honeybee products for 7 days following CCl4 injection. Development of liver damage and oxidative stress were monitored by measuring the activities of the enzymes alanine transaminase, aspartate transaminase, malondialdehyde, superoxide dismutase, and catalase. Antioxidant capacities of the bee products were identified using FRAP and DPPH assays, as well as by measuring total phenolic and flavonoid contents. RESULTS: The antioxidant activities of the honeybee products were highest in propolis, followed, in order, by pollen, honey, and royal jelly. Despite their different levels of antioxidant capacity, their roles in the prevention of liver damage induced by CCl4 were very similar, which can be explained through their bioavailability to the treated animals. CONCLUSIONS: Our results suggest that honey, propolis, pollen, and royal jelly significantly enhanced the healing of CCl4-induced liver damage, partially due to their antioxidant properties and bioavailability.

A comparative study of the antihyaluronidase, antiurease, antioxidant, antimicrobial and physicochemical properties of different unifloral degrees of chestnut (Castanea sativa Mill.) honeys.

This study was planned to investigate some physicochemical and anti-inflammatory, antioxidant, antimicrobial properties of three different degrees of unifloral characters of chestnut honeys. Antihyaluronidase, antiurease and antimicrobial activities were evaluated as anti-inflammatory characteristics. Total phenolic contents, flavonoids, tannins, phenolic profiles, ferric-reducing antioxidant power (FRAP), scavenging activities of 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS+) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals were evaluated as antioxidant properties. Color, optical rotation, conductivity, moisture, pH and ash content were evaluated as physicochemical parameters, and some sugars content, prolin, diastase, HMF and minerals (Na, K, Ca, P, Fe, Cu and Zn) were evaluated as chemical and biochemical parameters. All studied physicochemical and biological active properties were changed in line with the unifloral character of the chestnut honeys. A higher unifloral character was found associated with greater apitherapeutic capacity of the honey, as well as biological active compounds.

Inhibition properties of propolis extracts to some clinically important enzymes.

The present study was conducted to envisage inhibition effects of propolis on the crucial enzymes, urease, xanthine oxidase (XO) and acetylcholinesterase (AChE). Some of the antioxidant properties of the propolis samples were determined using the total phenolic content (TPE) and total flavonoids in the eight different ethanolic propolis extracts (EPE) samples. Inhibition values of the enzymes were expressed as inhibition concentration (IC50; mg/mL or µg/mL) causing 50% inhibition of the enzymes with donepezil, acetohydroxamic acid and allopurinol as reference inhibitors. All the propolis extracts exhibited variable inhibition effects on these enzymes, but the higher the phenolic contents the lower the inhibitions values (IC50 = 0.074 to 1.560 mg/mL). IC50 values of the P5 propolis sample having the highest TPE, obtained from Zonguldak, for AChE, urease and XO were 0.081 ± 0.009, 0.080 ± 0.006 and 0.074 ± 0.011 µg/mL, respectively. The EPE proved to be a good source of inhibitor agents that can be used as natural inhibitors to serve human health.

Diagnosis of Acute Coronary Syndrome with a Support Vector Machine.

Acute coronary syndrome (ACS) is a serious condition arising from an imbalance of supply and demand to meet myocardium's metabolic needs. Patients typically present with retrosternal chest pain radiating to neck and left arm. Electrocardiography (ECG) and laboratory tests are used indiagnosis. However in emergency departments, there are some difficulties for physicians to decide whether hospitalizing, following up or discharging the patient. The aim of the study is to diagnose ACS and helping the physician with his decisionto discharge or to hospitalizevia machine learning techniques such as support vector machine (SVM) by using patient data including age, sex, risk factors, and cardiac enzymes (CK-MB, Troponin I) of patients presenting to emergency department with chest pain. Clinical, laboratory, and imaging data of 228 patients presenting to emergency department with chest pain were reviewedand the performance of support vector machine. Four different methods (Support vector machine (SVM), Artificial neural network (ANN), Naïve Bayes and Logistic Regression) were tested and the results of SVM which has the highest accuracy is reported. Among 228 patients aged 19 to 91 years who were included in the study, 99 (43.4 %) were qualified as ACS, while 129 (56.5 %) had no ACS. The classification model using SVM attained a 99.13 % classification success. The present study showed a 99.13 % classification success for ACS diagnosis attained by Support Vector Machine. This study showed that machine learning techniques may help emergency department staff make decisions by rapidly producing relevant data.

A Member of Complementary Medicinal Food: Anatolian Royal Jellies, Their Chemical Compositions, and Antioxidant Properties.

This study investigated various chemical and antioxidant properties of Anatolian royal jelly samples. Moisture, pH, total protein, 10-hydroxy-2-decenoic acid (10-HDA) and sugars were analyzed from 18 samples. Total phenolic contents, ferric reducing antioxidant capacity and 2,2-diphenyl-1-picryhydrazyl (DPPH) free radical scavenging activity were measured as antioxidant determinants. 10-HDA contents and total protein content of fresh weight ranged between 1.0% and 3.9%, and 11.4% and 15.8%, respectively. The main sugars detected were glucose and fructose. Maltose, trehalose, and melibiose were detected at less than 1.0% in all samples. Lactose, a milk sugar, was detected in only 3 samples, at values between 0.8% and 1.4%. Total henolic content ranged from 91.0 to 301.0 mg gallic acid equivalents/kg fresh weight. Antioxidant activity is due to both to the total phenolic content, proteins and fatty acids of royal jelly. Anatolian royal jelly samples were not different from other royal jelly samples from across the world.

Effects of Mad Honey on Some Biochemical Parameters in Rats.

The aims of this study were to determine grayanotoxin (GTX-III) toxin level in mad honey using liquid chromatography-tandem mass spectrometry and examine the dynamic changes of certain biochemical parameters in blood serum of rats that consumed mad honey. For the experimental animal study, 20 Sprague-Dawley female rats were divided into 5 groups of 4 rats each, with one group being the control group (Group 1) and the others being the experimental groups (Groups 2-5). Groups 2, 3, 4, and 5 were, respectively, given mad honey extract at doses of 0.3, 0.6, 1.2, and 2.4 mg/g body weight/day via oral gavage for 8 days. According to results, the quantity of GTX-III found in the honey sample as 39.949 ± 0.020 µg GTX-III/g honey, and the biochemical analysis of the tested parameters (aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, alkaline phosphatase, creatine kinase, and creatine kinase muscle and brain) showed a significant elevation with increasing concentration of honey. In conclusion, the use of increasing concentrations of Rhododendron honey was seen as a source of enzymatic symptoms.

Honey shows potent inhibitory activity against the bovine testes hyaluronidase.

The purpose of this study was to investigate the anti-hyaluronidase activities of honeys from different botanical origins honeys in order to determine their anti-inflammatory properties. The total phenolic contents, total flavonoids and total tannin levels of six types of honey, chestnut, oak, heather, pine, buckwheat and mixed blossom, were determined. Concentration-related inhibition values were tested turbidimetrically on bovine testis hyaluronidase (BTHase) as IC50 (mg/mL). All honeys exhibited various concentration-dependent degrees of inhibition against BTHase. Inhibition values varied significantly depending on honeys' levels of phenolic contents, flavonoid and tannin. The honeys with the highest anti-hyaluronidase activity were oak, chestnut and heather. In conclusion, polyphenol-rich honeys have high anti-hyaluronidase activity, and these honeys have high protective and complementary potential against hyaluronidase-induced anti-inflammatory failures.

An investigation of Turkish honeys: their physico-chemical properties, antioxidant capacities and phenolic profiles.

This study investigated some physico-chemical and biochemical characteristics of different honey types belonging to Turkish flora. Sixty-two honey samples were examined on the basis of pollen analyses, including 11 unifloral honeys (chestnut, heather, chaste tree, rhododendron, common eryngo, lavender, Jerusalem tea, astragalus, clover and acacia), two different honeydew honeys (lime and oak), and 7 different multifloral honeys. Electrical conductivity, moisture, Hunter color values, HMF, proline, diastase number, and sugar analyses of the honey samples were assessed for chemical characterization. Some phenolic components were analyzed by reverse phase high performance liquid chromatography (RP-HPLC) to determine honeys' phenolic profiles. Total phenolic compounds, total flavonoids, ferric reducing antioxidant capacity (FRAP) and 2,2-diphenyl-1-picryhydrazyl (DPPH) free radical scavenging activity were measured as antioxidant determinants. The study results confirm that physico-chemical and biological characteristics of honeys are closely related to their floral sources, and that dark-colored honeys such as oak, chestnut and heather, have a high therapeutic potential.

Polyphenol oxidase activity and antioxidant properties of Yomra apple (Malus communis L.) from Turkey.

In this study, firstly, antioxidant and polyphenol oxidase (PPO) properties of Yomra apple were investigated. Seventeen phenolic constituents were measured by reverse phase-high-performance liquid chromatography (RP-HPLC). Total phenolic compounds (TPCs), ferric reducing antioxidant power (FRAP) and 2, 2-diphenyl-1-picrylhydrazyl radical (DPPH) scavenging activities were performed to measure antioxidant capacity. Some kinetic parameters (Km, Vmax), and inhibition behaviors against five different substrates were measured in the crude extract. Catechin and chlorogenic acid were found as the major components in the methanolic extract, while ferulic acid, caffeic acid, p-hydroxybenzoic acid, quercetin and p-coumaric acid were small quantities. Km values ranged from 0.70 to 10.10 mM in the substrates, and also 3-(4-hydroxyphenyl) propanoic acid (HPPA) and L-DOPA showed the highest affinity. The inhibition constant of Ki were ranged from 0.05 to 14.90 mM against sodium metabisulphite, ascorbic acid, sodium azide and benzoic acid, while ascorbic acid and sodium metabisulphite were the best inhibitors.

Hepatoprotective potential of chestnut bee pollen on carbon tetrachloride-induced hepatic damages in rats.

Bee pollen has been used as an apitherapy agent for several centuries to treat burns, wounds, gastrointestinal disorders, and various other diseases. The aim of our study was to investigate the hepatoprotective effects of chestnut bee pollen against carbon tetrachloride (CCI4)-induced liver damage. Total phenolic content, flavonoid, ferric reducing/antioxidant power, and DPPH radical activity measurements were used as antioxidant capacity determinants of the pollen. The study was conducted in rats as seven groups. Two different concentrations of chestnut bee pollens (200 and 400 mg/kg/day) were given orally and one group was administered with silibinin (50 mg/kg/day, i.p.) for seven days to the rats following the CCI4 treatment. The protective effect of the bee pollen was monitored by aspartate transaminase (AST) and alanine transaminase (AST) activities, histopathological imaging, and antioxidant parameters from the blood and liver samples of the rats. The results were compared with the silibinin-treated and untreated groups. We detected that CCI4 treatment induced liver damage and both the bee pollen and silibinin-treated groups reversed the damage; however, silibinin caused significant weight loss and mortality due, severe diarrhea in the rats. The chestnut pollen had showed 28.87 mg GAE/g DW of total phenolic substance, 8.07 mg QUE/g DW of total flavonoid, 92.71 mg Cyn-3-glu/kg DW of total anthocyanins, and 9 mg ß -carotene/100 g DW of total carotenoid and substantial amount of antioxidant power according to FRAP and DPPH activity. The results demonstrated that the chestnut bee pollen protects the hepatocytes from the oxidative stress and promotes the healing of the liver damage induced by CCI4 toxicity. Our findings suggest that chestnut bee pollen can be used as a safe alternative to the silibinin in the treatment of liver injuries.

Inhibition of carbonic anhydrase isozymes I and II with natural products extracted from plants, mushrooms and honey.

Different natural products and secondary metabolites from mushrooms, teas, honeys, mosses, plants and seaweeds were investigated for their in vitro inhibitory effects on human carbonic anhydrase (hCA, E.C.4.2.1.1) isoforms I and II. Inhibition data were correlated with the total phenol content in the extract and investigated with the pure compounds believed to be responsible for this activity. Methanolic extracts were prepared for 17 such pure chemicals present in the natural products and for 41 diverse natural products. The IC(50) values were in the range of 0.11-66.50 µg/mL against hCA I and of 0.09-54.54 µg/mL against hCA II, respectively. The total phenol content was in the range of 0.02-1318.96 (as milligrams of gallic acid equivalents) per gram of sample. These data offer new insights on possible novel classes of CA inhibitors based on natural products, possessing a range of chemical structures not present in the classical inhibitors with pharmacological applications, such as the sulfonamides and sulfamates.

Effects of forest fire on soil nutrients in Turkish pine (Pinus brutia, Ten) ecosystems.

Fire is a long-standing and poorly understood component of the Mediterranean forestlands in Turkey. Fire can alter plant composition, destroy biomass, alter soil physical and chemical properties and reduce soil nutrient pools. However fire can also promote productivity of certain ecosystems by mineralizing soil nutrients and promoting fast growing nitrogen fixing plant species. Fire effects on soils and ecosystems in Turkey and Mediterranean regions are not well understood. This study uses a retrospective space-for-time substitution to study soil macro-nutrient changes on sites which were burned at different times during the last 8 years. The study sites are in the Fethiye Forest Management Directorate in the western Mediterranean Sea region of Turkey. Our samples show 40% less Soil C, and cation exchange capacity (CEC) at 0-20 cm soil depth two weeks after the fire. Soil C and CEC appear to recover to pre-fire level in one year. Concentrations of Mg were significantly lower on new-burn sites, but returned to pre-fire levels in one year. Total soil N concentrations one and two years after fire were 90% higher than other sites, and total P was 9 times higher on new-burn site than averages from other sites. Some implications of these results for forest managers are discussed.

Effects of different pretreatments on germination of Prunus serotina seed sources.

Establishing intensive plantations of fast growing hardwood tree species that have high market values in the forest industry can narrow the gap between Turkey's demand and the supply of quality hardwood products. Black cherry (P. serotina Ehrh.) is a fast growing hardwood species with a high market value. Introducing and intensively growing black cherry (BC) in Turkey may significantly reduce the country's quality wood shortage. Adequate seed germination constitutes the first essential step for successful establishments. In this paper effects of different pretreatments, including artificial and natural stratification, on the seeds of different BC seed sources (SSs) were studied. Pretreatments had substantial effects on the dormancy breaking and germination behaviours of the SSs. Consecutive periods of complex warm and cold artificial stratification regimes longer than 90 days or natural stratification (where seeds were assumed to be naturally exposed to this complexity) resulted in best dormancy breaking and, in turn, germination among all pretreatments. Deeper dormancy and reduced germination rates of some BC seeds as the altitude of the source increases might suggest an ecological adaptive strategy of the species. BC may have deeper morphophysiological dormancy than is commonly believed. Seed size may have a positive effect on seed germination.