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Identification of nitric oxide metabolites in various honeys: effects of intravenous honey on plasma and urinary nitric oxide metabolites concentrations.

Honey has antibacterial activity, promotes healing, and enhances immunity. Its acidity, osmotic effects of its high content of sugar, and hydrogen peroxide are assumed to be responsible for its effects. In this study, various honeys were investigated for the presence of nitrite/nitrate, the stable nitric oxide (NO) metabolites, and the effects of intravenous infusion of honey on urinary and plasma NO end products were studied in healthy sheep. Seven kinds of honey, different in their origin (three from Yemen, two from the United Arab Emirates, one from Germany, and one from India), color, and duration of storage, were investigated for the presence of NO metabolites. The assessment of NO metabolites was performed before and after exposure of the honey samples to heating (80 degrees C for 1 hour) or ultraviolet light (for 24 hours). Seven healthy male sheep were used for the study. Fresh unprocessed yellow honey (2 g/kg of body weight) was infused over a period of 45 minutes to each fasting sheep. Plasma and urinary NO metabolites were measured before and after the infusion. All the honey samples examined had various concentrations of NO metabolites; the highest concentration was in the fresh dark honey collected from Yemen, and the lowest in 1-year-stored dark honey collected from India. Darker or fresh honeys contained more NO metabolites than light or stored honey. After heating, NO metabolites decreased in all the kinds of honey. After ultraviolet exposure, NO metabolites were decreased in four kinds of honey, increased in one kind, and unchanged in two kinds. The darker stored honey had more resistance to heating and ultraviolet exposure. Intravenous infusion of honey elevated urinary NO metabolites from 8.4 +/- 7.4 micromol/L to 14.9 +/- 10 micromol/L during the first 60-90 min after infusion and to 35.2 +/- 34 micromol/L during the next 150-180 min. Plasma NO metabolites were increased during 1, 2, and 3 hours after infusion by 3%, 3.6%, and 17%, respectively. No side effects were reported with the use of intravenous honey. It might be concluded that honey contains various concentrations of NO metabolites. Its intravenous infusion increased plasma and urinary NO metabolites. It is assumed that NO might be responsible, in part, for the biological and therapeutic effects of honey.

Analysis of Variance↗

Allergy to honey: relation to pollen and honey bee allergy.

To identify the allergenic components of honey we studied 22 patients with a history of systemic allergic symptoms following honey ingestion. The group of honey-allergic patients was compared with three control groups: 10 subjects sensitized to artemisia, 10 with honey bee venom allergy and 10 without a history of atopy or bee sting reactions. The allergological tests included skin tests and RAST with three different kinds of Swiss honey (dandelion, forest and rape), pollen of compositae species, celery tuber, extract of bee pharyngeal glands, honey bee venom and bee whole body extract. The results show that 3/4 of honey-allergics are sensitive to dandelion honey and 13 of 22 also to compositae pollen. Nine of the honey allergic patients were sensitized to honey bee venom, 3 also to bee pharyngeal glands and to bee whole body extract. Analysis of diagnostic tests and RAST inhibition studies suggest that besides compositae pollen other allergens, most likely of bee origin are important. In honey allergics primary sensitization may be due either to the honey itself, to airborne compositae pollen or even to cross-reacting bee venom components.

Adolescent↗

From the linden flower to linden honey--volatile constituents of linden nectar, the extract of bee-stomach and ripe honey.

Honey is produced by honeybees (Apis mellifera), which collect nectar from flowers, digest it in their bodies, and deposit it in honeycombs, where it develops into ripe honey. We studied the evolution of the volatile constituents from the nectar of linden blossoms (Tilia cordata) to honey via the 'intermediate' honeybee. The sampling of the contents of the honey stomach or honey sack of the bee is unique. Extracts were prepared from nectar, from the liquid of the honey stomach, and from ripe honey. The chemistry is extremely complex, and compounds spanning from monoterpenes (hydrocarbons, ethers, aldehydes, acids, and bifunctional derivatives), isoprenoids, aromatic compounds (phenylpropanoids, phenols), and products degraded from fatty acids to alkaloids, were identified. Some compounds definitely stem from the plants, whereas other interesting constituents can be attributed to animal origin. Two derivatives of decanoic acid, 9-oxodec-2-enoic acid (12) and 9-hydroxydec-2-enoic acid, identified in the honey are known to be constituents of the so-called 'Queen's pheromone'. Two metabolites of these acids were identified in the extract of the honey stomach: 8-oxononanal (10), a new natural product, and 8-oxononanol (11). There structures were confirmed by synthesis. Nectar and honey stomach contain many aldehydes, which, due to the highly oxidative atmosphere in the honeycomb, are found as corresponding acids in the honey. Two acids were newly identified as 4-isopropenylcyclohexa-1,3-diene-1-carboxylic acid (14) and 4-(1-hydroxy-1-methylethyl)-cyclohexa-1,3-diene-1-carboxylic acid (15).

Animals↗

Food allergy to honey: pollen or bee products? Characterization of allergenic proteins in honey by means of immunoblotting.

OBJECTIVE: To characterize the allergenic components of honey, 23 patients allergic to honey were investigated. All displayed allergic symptoms after ingestion of honey or honey-containing products, ranging from itching in the oral mucosa to severe systemic symptoms to anaphylactic shock. METHODS AND RESULTS: Immunoblot analyses of the patients' sera revealed IgE binding to proteins at a molecular mass of 54 kd, 60 kd, 72 kd, or to a 30 kd/33 kd double band, or to both in sunflower honey extracts. The three bands corresponding to higher molecular mass proteins could also be detected in the three other kinds of honey (locust tree, European chestnut and forest honey) that were tested and represented bee products because IgE binding to these proteins was inhibited by extracts of honeybee heads and extracts of isolated bee venom sacs. The 30 kd/33 kd bands could be identified as sunflower honey-specific. When testing sera from patients allergic to bee venom with honey extracts, in seven of 10 cases IgE binding to bee-specific components could be observed. CONCLUSION: Both proteins derived from secretions of pharyngeal and salivary glands of honeybee heads and pollen proteins contained in the honey cause allergic reactions to honey.

Adolescent↗

Cyanide residue levels in extracted honey, comb honey and wax cappings.

Cyanide (CN) residue levels were determined in samples of extracted honey, comb honey and was cappings at 1 hr, 24 hr, and 48 hr intervals after destroying the bees in honey bee colonies with normal (ca. 8.5 g) and twice normal (ca. 17 g) doses of CyanogasR A-dust. Applications of CyanogasR A-dust, administered by means of a dust pump at normal and twice normal doses, gave an average residue of 0.01 and 0.04 microgram CN/g of extracted honey, 0.01 and 0.02 microgram CN/g of comb honey and 0.04 and 0.06 microgram CN/g of wax cappings, respectively. When the CyanogasR A-dust (ca. 17 g) was placed on a tray and placed on the bottom board of the hive, the average residue levels for extracted honey, comb honey and wax cappings were less than 0.004, 0.01 and 0.02 microgram CN/g, respectively. Random honey samples from beekeepers, who used CyanogasR to destroy bees, had a median level of 0.031 microgram CN/g, whereas honey from a packing plant and other commercial samples contained less than 0.004--0.026, median less than 0.004 microgram CN/g. Based on residue data from this study, the temporary registration for CyanogasR, to kill honey bees after crop removal, was revised to a full registration in May 1977.

Bees↗

A comparison of the sensitivity of wound-infecting species of bacteria to the antibacterial activity of manuka honey and other honey.

Both honey and sugar are used with good effect as dressings for wounds and ulcers. The good control of infection is attributed to the high osmolarity, but honey can have additional antibacterial activity because of its content of hydrogen peroxide and unidentified substances from certain floral sources. Manuka honey is known to have a high level of the latter. Seven major wound-infecting species of bacteria were studied to compare their sensitivity to the non-peroxide antibacterial activity of manuka honey and to a honey in which the antibacterial activity was primarily due to hydrogen peroxide. Honeys with activity in the middle of the normal range were used. A comparison of the median response of the various species of bacteria showed no significant difference between the two types of activity overall, but marked differences between the two types of activity in the rank order of sensitivity of the seven bacterial species. The non-peroxide antibacterial activity of manuka honey at a honey concentration of 1.8% (v/v) completely inhibited the growth of Staphylococcus aureus during incubation for 8 h. The growth of all seven species was completely inhibited by both types of honey at concentrations below 11% (v/v).

Bacteria↗

Could honey have a place in colitis therapy? Effects of honey, prednisolone, and disulfiram on inflammation, nitric oxide, and free radical formation.

BACKGROUND/AIMS: The purpose of this study was to investigate the potential therapeutic roles of honey, prednisolone and disulfiram in an experimental model of inflammatory bowel disease. Another aspect of the study was to find out whether these substances have any effect on nitric oxide (NO) and free radical production. METHODS: After the induction of colitis with trinitrobenzene sulfonic acid in 64 male rats, physiological saline, honey, prednisolone and disulfiram enemas were applied to the rats once daily for 3 days (acute treatment groups) or 7 days (chronic treatment groups). Control groups received only saline enemas. Rats were killed on the 4th or 8th days and their colonic mucosal damage was quantitated using a scoring system. Acute and chronic inflammatory responses were determined by a mucosal injury score, histological examination and measurement of the myeloperoxidase (MPO) activity of tissues. The content of malonylaldehyde (MDA) and NO metabolites in colon homogenates was also measured to assess the effects of these substances on NO and free oxygen radical production. RESULTS: Estimation of colonic damage by mucosal injury scoring was found to be strongly correlated with the histologic evaluation of colon specimens. On the other hand, mucosal injury scores were not correlated with MPO, MDA or NO values. There were significant differences between the MPO results of chronic-control and chronic-honey groups, as well as chronic-control and chronic-prednisolone groups (p = 0.03 and p = 0.0007). The acute honey, prednisolone, and disulfiram groups had significantly lower MDA results compared to the acute control group (p = 0.04, p = 0.02, and p = 0.04). In terms of NO, there was no significant difference between the treatment and control groups. NO was found to have a strong relationship with MDA (p = 0.03) and MPO values (p = 0.001). On the other hand, MPO results were not found to be correlated with MDA values (p > 0.05). CONCLUSIONS: MPO activity is not directly proportional to the severity of the inflammation, but it may only determine the amount of neutrophil in the tissues. Inflammatory cells are not the sole intensifying factor in colitis. Therefore, mucosal injury scores may not correlate well with MPO activities. In an inflammatory state NO and MPO levels have a strong relationship, since NO is released from the neutrophils. In an inflammatory model of colitis, intrarectal honey administration is as effective as prednisolone treatment. Honey may have some features in the treatment of colitis, but this issue requires further investigation. Honey, prednisolone and even disulfiram also have some value in preventing the formation of free radicals released from the inflamed tissues. Prednisolone may also have some possible benefits in the inhibition of NO production in colitis therapy.

Animals↗

The sterilization of honey with cobalt 60 gamma radiation: a study of honey spiked with spores of Clostridium botulinum and Bacillus subtilis.

Unprocessed honey is a recognized wound-healing remedy. However, to make clinical use of honey acceptable, it should be sterile. To find the lowest dose of irradiation needed for sterilization, six batches of honey (a-f) were gamma irradiated with 6, 12, 18, 22 and 25 kGy Cobalt-60. After a dose of 25 kGy the antibacterial activity was not altered. Presumably glucose oxidase (EC 1.1.3.4), which produces hydrogen peroxide, is not easily damaged by irradiation. Amylase activity on the other hand was significantly reduced to 19%, 19%, 21%, 22%, 43% in batches a), b), c), d) and f) respectively, whereas no decrease was observed in batch e). All batches spiked with approximately 10(6) spores from Cl. botulinum or B. subtilis per 50 g honey proved to be sterile after irradiation with a dose of 25 kGy. Honey was also spiked with Cl. botulinum at up to 5000 spores per 50 g honey, which is the upper limit of natural contamination. The sterilizing dose in this case was 18 kGy.

Anti-Bacterial Agents↗

Usefulness of amino acid composition to discriminate between honeydew and floral honeys. Application to honeys from a small geographic area.

With the aim of finding methods that could constitute a solid alternative to melissopalynological and physicochemical analyses to determine the botanical origin (floral or honeydew) of honeys, the free amino acid content of 46 honey samples has been determined. The honeys were collected in a small geographic area of approximately 2000 km(2) in central Spain. Twenty-seven honey samples were classified as floral and 19 as honeydew according to their palynological and physicochemical analyses. The resulting data have been subjected to different multivariant analysis techniques. One hundred percent of honey samples have been correctly classified into either the floral or the honeydew groups, according to their content in glutamic acid and tryptophan. It is concluded that free amino acids are good indicators of the botanical origin of honeys, saving time compared with more tedious analyses.

Amino Acids↗

Improved method for extraction and LC-MS analysis of pyrrolizidine alkaloids and their N-oxides in honey: application to Echium vulgare honeys.

A method for analyzing honey samples was developed that enabled the simultaneous detection and identification of pyrrolizidine alkaloids and their N-oxides. Honey samples were treated with methanol or dilute sulfuric acid and then centrifuged to remove insoluble material. Subsequent strong cation exchange, solid-phase extraction of the supernatant provided a fraction that was analyzed for the presence of pyrrolizidine alkaloids and their N-oxides using high-pressure liquid chromatography coupled to electrospray ionization mass spectrometry. The procedure was validated using extracts of Echium plantagineum and authenticated standards of pyrrolizidine alkaloids and their N-oxides from other plant sources. Of several variations of the solid-phase extraction method assessed in this study, the best combination for generic use involved the dilution of honey with 0.05 M sulfuric acid and the subsequent application of the centrifuged solution to solid-phase extraction columns at the rate of a maximum of 10 g of honey per solid-phase extraction column. The method was applied to the analysis of nine floral honeys, five of which were attributed by the apiarist to Echium vulgare. Seven of the honey samples were positive for pyrrolizidine alkaloids and N-oxides characteristic of E. vulgare.

Chromatography, High Pressure Liquid↗

Honey revisited: a reappraisal of honey in pre-industrial diets.

In pre-industrial times, honey was the main source of concentrated sweetness in the diets of many peoples. There are no precise figures for per capita consumption during most periods in history because honey was part of either a hunter-gatherer or subsistence economy. Until now, historians and food writers have proposed that it was a scarce commodity available only to a wealthy few. We do know, however, that in a cash economy honey was sold in large units (gallons and even barrels) and it was present in such abundance that mead, made from honey, was a common alcoholic drink. A reappraisal of the evidence from the Stone Age, Antiquity, the Middle Ages and early Modern times suggests that ordinary people ate much larger quantities of honey than has previously been acknowledged. Intakes at various times during history may well have rivalled our current consumption of refined sugar. There are implications therefore for the role of sugar in modern diets. Refined sugar may not have displaced more nutrient-rich items from our present-day diets but only the nutritionally comparable food, honey.

Diet↗

A survey of Nosema apis of honey bees (Apis mellifera L.) producing the famous Anzer honey in Turkey.

The aim of this study is to find out the ratio of Nosema infected honey bees which are producing the famous Anzer honey that is used for the cure of the illnesses such as farangitis, tonsilitis, ulceration, and scratchs due to the experiences of the people living in Turkey. Honey bee samples were collected from two different regions of Anzer plain in July. Honey bee abdomens were homogenized and 1 ml distilled water was added for each honey bee. Later, 0.1 ml out of this solution was examined by Neubauer slides and the number of Nosema apis spores were counted. The results showed that Nosema apis significantly infected the honey bees although it was summer season. However, the summer season at the Anzer plain, when compared with the Mediterranean climate, is considered to be spring.

Animals↗

Honey protein as internal standard for stable carbon isotope ratio detection of adulteration of honey.

Using the difference in stable carbon isotope ratio between a honey and its protein fraction permits objective evaluation of possible adulteration of honey with small amounts (7-20%) as well as larger amounts of corn or cane sugar. The present uncertainty in interpretation of results from pure honey with delta 13C values outside the generally accepted limits for pure honey of -27.5% to -23.5% is eliminated; likewise TLC testing to resolve questionable samples with delta 13C values between -23.5 and -21.5% is not needed. Fifty certified samples of pure honey were used to establish criteria for purity, and 38 other samples with delta 13C values in the "questionable" or "adulterated" range for the AOAC official method were tested. A difference of 1.0% or more between honey and protein fractions is proposed to indicate adulteration.

Carbon Isotopes↗

Use of differential scanning calorimetry (DSC) as a new technique for detection of adulteration in honeys. 1. Study of adulteration effect on honey thermal behavior.

Differential scanning calorimetry (DSC) was used to study the thermal behavior of authentic honeys (Lavandula, Robinia, and Fir honeys) and industrial sugar syrups. Thermal or thermochemical parameters such as the glass transition temperature (Tg), enthalpies of fusion (DeltaH(fus)), and heat capacity variation (DeltaC(p)) were measured. The syrups and honeys showed significant differences in thermal phenomena, as well as in their amplitude and position on the temperature scale. Results showed good reproducibility of the method for all samples studied. The effect of adulteration of honey with different amounts of syrup (5, 10, 20, 40, and 60%) was investigated. A linear relationship was found between the percentage of added syrup and the glass transition temperature. A similar relationship was obtained from the enthalpy of fusion results in the temperature range of 40-90 degrees C. Under applied conditions, the effects of adulteration of honeys by industrial syrups appeared to be detectable from a level as low as 5%.

Calorimetry, Differential Scanning↗

Honey characterization and adulteration detection by pattern recognition applied on HPAEC-PAD profiles. 1. Honey floral species characterization.

An improved COFRAC (COmité FRançais d'ACréditation) method for the analysis and evaluation of the quality of honey by high-performance anion-exchange chromatography of sugar profiles is proposed. With this method, both minor and major sugars are simultaneously analyzed and the technique is integrated in a new chemometric approach, which uses the entire chromatographic sugars profile of each analyzed sample to characterize honey floral species. Sixty-eight authentic honey samples (6 varieties) were analyzed by high-performance anion-exchange chromatography-pulsed amperometric detection. A new algorithm was developed to create automatically the corresponding normalized data matrix, ready-to-use in various chemometric procedures. This algorithm transforms the analytical profiles to produce the corresponding calibrated table of the surfaces or intensities according to retention times of peaks. The possibility of taking into account unknown peaks (those for which no standards are available) allows the maximum chemical information provided by the chromatograms to be retained. The parallel application of principal component analysis (PCA)/linear discriminant analysis (LDA) and artificial neural networks (ANN) shows a high capability in the classification of the analyzed samples (LDA, 93%; ANN, 100%) and a very good discrimination of honey groups. This work is the starting point of the elaboration of a new system designed for the automatic pattern recognition of food samples (first application on honey samples) from chromatographic analyses for food characterization and adulteration detection.

Algorithms↗

Protein analysis of honeys by fast protein liquid chromatography: application to differentiate floral and honeydew honeys.

Fast protein liquid chromatography on a Superdex 75 HR column has been applied to analyze the proteins of 29 honeys, 12 of floral origin and 17 from honeydew. The molecular masses were comprised between 13100 and 94000 Da. Seven peaks have been separated; four of them were present in all of the honeys, and three were only present in some honeys. Direct observation of the chromatograms of the floral and honeydew honeys did not reveal any information about their botanical origins. However, both types of honeys can be distinguished with the percentages of the areas of four of the seven chromatographic peaks obtained.

Amino Acids↗

[The pros and cons of honey--are statements about the effects of honey "scientifically verified"?].

The physiological effects attributed to honey are described and discussed. These effects are to a great extent generalizations from experimental and clinical observations. In some cases the effects attributed to honey are based on tests which are no longer acceptable. Most of the statements about physiological effects caused by eating honey cannot be supported by scientific methods. Under German food law these are designated as "misleading". Honey is comparable to sugar in its nutritive value. Other nutrients like protein, minerals and vitamins are present in such small amounts that they have no nutritional importance.

Acetylcholine↗

[Proteins in honey. II. Gel-chromatography, enzymatic acitivity and origin of honey-proteins (author's transl)].

The honey proteins were separated into five distinct peaks by freezing and thawing and subsequent chromatography on Sephadex G-200. The enzymatic activities were always located in the same peaks and clearly separated. The elution curves of the individual honeys were very similar. No significant differences were observed neither among floral honeys not between floral and honeydew honeys. The determination of the elution coefficients, however, showed that three of the five peaks originate in the bee the other two in the plant.

Chromatography, Gel↗