Genistein Supplementation and Bone Health in Breast Cancer in Rats.

The aim of our study was to analyse the effect of supplementation with various forms of genistein (nano-, micro-, and macro-) on the mineral status of rat femurs in conditions of DMBA-induced mammary gland neoplasia. Thirty-two 30-day-old Sprague Dawley rats were used in the study. The rats were divided into four experimental groups: a control group (without supplementation) and groups supplemented with nanosized (92 ± 41 nm), microsized (587 ± 83 nm), and macrosized genistein. Micromorphometric and histological examination of the rat femurs were performed, as well as analysis of the weight and mineral composition (17 elements). Quadrupole ICP-MS was used for analysis of all trace elements. Supplementation with genistein (nano-, micro-, and macro-) was shown to cause changes in the mineral composition of the bones. In the rats receiving nanogenistein, disintegration of the bone tissue was observed. The femurs of these animals had higher content of calcium (by nearly 300%) and potassium (by 25%) than the other groups, while the level of magnesium was about 22% lower. In the case of microelements, there were increases in copper (by 67%), boron (48%), manganese (13%), and nickel (100%), and a 16% decrease in strontium compared to the bones of rats without genistein supplementation. Changes in micromorphometric parameters, resulting in increased bone fragility, were observed. Administration of genistein was found to have an effect on the amount of trace elements in the bone tissue of rats with breast cancer.

Effect of Nano- and Microzinc Supplementation on the Mineral Composition of Bones of Rats with Induced Mammary Gland Cancer.

BACKGROUND: The aim of this study was to determine changes in the mineral composition of the bones of rats with chemically induced mammary gland cancer and to attempt to establish whether a specific diet modification involving the inclusion of zinc ions in two forms-nano and micro-will affect the mineral composition of the bones. METHODS: Female Sprague-Dawley rats were used for the research. The animals were randomly assigned to three experimental groups. All animals were fed a standard diet (Labofeed H), and selected groups additionally received zinc nanoparticles or microparticles in the amount of 4.6 mg/mL. To induce mammary cancer, the animals were given 7,12-dimethyl-1,2-benz[a]anthracene. The content of Ag, As, B, Ba, Cd, Cr, Cu, Mn, Ni, Pb, Rb, Se, Sr, Tl, U, and V was determined using ICP-MS, while that of Ca, Fe, K, Mg, Na, and Zn was determined using FAAS. RESULTS: The use of a diet enriched with zinc nano- or microparticles significantly influenced the content of the elements tested. In the bones of rats fed a diet with zinc nanoparticles, changes were found in the content of Ca, Mg, Zn, Cd, U, V, and Tl, while in the case of the diet supplemented with zinc microparticles, there were differences in six elements-Ca, Mg, B, Cd, Ag, and Pb-compared to animals receiving an unsupplemented diet. CONCLUSIONS: The content of elements in the bone tissue of rats in the experimental model indicates disturbances of mineral metabolism in the tissue at an early stage of mammary cancer.

Phytochemical screening and effect of Viscum album L. on monoamine oxidase A and B activity and serotonin, dopamine and serotonin receptor 5-HTR1A levels in Galleria mellonealla (Lepidoptera).

ETHNOPHARMACOLOGICAL RELEVANCE: Viscum album L. (European mistletoe), a member of the Santalaceae, is a hemiparasitic, evergreen shrub growing on deciduous and coniferous trees. In traditional and folk medicine, mistletoe was used for the treatment of central nervous system disorders such as epilepsy, hysteria, insomnia, nervous excitability, neuralgia, headache, dizziness and fatigue. However, relatively little is known of its neuropharmacological activity. AIM OF THE STUDY: The aim of the present study was to evaluate the effect of treatment with aqueous and hydroethanolic extracts from Viscum album L. parasitizing birch, linden and pine, on MAO-A and MAO-B activity as well as serotonin, dopamine and serotonin receptor 5-HTR1A levels in Galleria mellonella (Lepidoptera) larvae. MATERIALS AND METHODS: The phytochemical composition of the extracts was characterised using UPLC-DAD-ESI-MS/MS. To investigate the neuropharmacological activity of Viscum album L. extracts, Galleria mellonella (Lepidoptera) larvae were used as a model organism. The inhibitory potential of the extracts against MAO-A and MAO-B was determined by fluorometry. The serotonin, dopamine and serotonin receptor 5-HTR1A levels in larvae hemolymph after treatment were quantified by ELISA. RESULTS: UPLC-DAD-ESI-MS/MS analysis allowed the identification of 88 compounds, either full or in part. Most of the characterised phytochemicals were flavonoids, hydroxycinnamic acids and lignans. Screening found that aqueous and hydroethanolic mistletoe extracts inhibited the enzymatic activity of either MAO-A or MAO-B or both. Additionally, mistletoe extract administration increased the levels of serotonin and serotonin receptor 5-HTR1A. None of the tested extracts had any significant effect on dopamine level. CONCLUSIONS: A key novel finding was that the aqueous and hydroethanolic extracts from Viscum album L. inhibited monoamine oxidase activity and increased the levels of serotonin and serotonin receptor 5-HTR1A in Galleria mellonella (Lepidoptera) larvae. These properties may be due to the presence of phenolic constituents, particularly flavonoids. Further research based on bioassay-guided fractionation of mistletoe is needed to identify CNS-active molecules.

Separation of menaquinone-7 geometric isomers by semipreparative high-performance liquid chromatography with silver complexation and identification by nuclear magnetic resonance.

Dietary supplements containing vitamin K2 are often used to prevent osteoporosis, vascular calcification and coronary heart disease. It has been shown that some of these products contain a mixture of menaquinone-7 geometric isomers. Since the geometric shape may influence biological activity, there was a need for a semipreparative method to isolate single compounds for further studies. Here, we present an argentation chromatographic method for the separation of menaquinone-7 isomers and an nuclear magnetic resonance (NMR) methodology for the configuration assignment of isoprenoid side chain. The DFT calculations were performed to determine more energetically favorable complexes between the cis or trans menaquinone-7 isomers and the silver cation. Seventeen components were resolved, and fractions were collected and subjected to NMR study. Structures and chemical shifts for thirteen new compounds were assigned, and the identity of three known compounds was confirmed.

Relationship between Structure and Biological Activity of Various Vitamin K Forms.

Vitamin K is involved many biological processes, such as the regulation of blood coagulation, prevention of vascular calcification, bone metabolism and modulation of cell proliferation. Menaquinones (MK) and phylloquinone vary in biological activity, showing different bioavailability, half-life and transport mechanisms. Vitamin K1 and MK-4 remain present in the plasma for 8-24 h, whereas long-chain menaquinones can be detected up to 96 h after administration. Geometric structure is also an important factor that conditions their properties. Cis-phylloquinone shows nearly no biological activity. An equivalent study for menaquinone is not available. The effective dose to decrease uncarboxylated osteocalcin was six times lower for MK-7 than for MK-4. Similarly, MK-7 affected blood coagulation system at dose three to four times lower than vitamin K1. Both vitamin K1 and MK-7 inhibited the decline in bone mineral density, however benefits for the occurrence of cardiovascular diseases have been observed only for long-chain menaquinones. There are currently no guidelines for the recommended doses and forms of vitamin K in the prevention of osteoporosis, atherosclerosis and other cardiovascular disorders. Due to the presence of isomers with unknown biological properties in some dietary supplements, quality and safety of that products may be questioned.

Biological activity of mistletoe: in vitro and in vivo studies and mechanisms of action.

Mistletoe has been used as treatment of many diseases in traditional and folk medicine. To date, anticancer, immunomodulatory, cardiac, antidiabetic, hepatoprotective, neuropharmacological, antibacterial and antifungal properties of mistletoe extracts have been studied the most. In this review, we summarized in vitro and in vivo studies on the pharmacological activity of Viscum species. Furthermore, we proposed the possible mechanisms of action of this herb, which might include many signalling pathways. Mistletoe could regulate either similar or different targets in various pathways that act on membrane receptors, enzymes, ion channels, transporter proteins and transcriptional targets. Still, pharmacological activities of mistletoe have been investigated mainly for crude extracts. It is a new field for scientists to determined which chemical compounds are responsible for the individual biological activities of mistletoe and how these activities are achieved. As a result, mistletoe might become a source of new complementary therapies supporting the treatment of many diseases.

Therapeutic potential of mistletoe in CNS-related neurological disorders and the chemical composition of Viscum species.

ETHNOPHARMACOLOGICAL RELEVANCE: Viscum album L., commonly known as mistletoe, has been used for centuries in traditional medicine to treat various neurological diseases, including epilepsy, hysteria, nervousness, hysterical psychosis, dizziness and headaches. AIM OF THE STUDY: The aim of this review is to summarize existing evidence confirming the influence of mistletoe on the central nervous system and to investigate the compounds that may be responsible for this activity. MATERIALS AND METHODS: Available information from studies of various species of the Viscum L. genus was collected from scientific journals, books, and reports via a library and an electronic data search (Elsevier, Google Scholar, PubMed, Springer, Science Direct, ResearchGate, and ACS). RESULTS: The main chemical constituents of Viscum L. species are viscotoxins, lectins, flavonoids, phenolic acids, terpenoids, sterols, phenylpropanoids, and alkaloids. Various extracts of Viscum album L. showed central nervous system activity, including antiepileptic, sedative, antipsychotic, anxiolytic, antidepressant and antinociceptive effects in mice and rats. Additionally, the extracts increased the level of brain-derived neurotrophic factor, prevented apoptotic neuronal death induced by amyloid ß and weakly inhibited cholinesterase activity. CONCLUSIONS: Numerous historical references describe the use of mistletoe for the treatment of central nervous system disorders. In recent years, studies have started to confirm the antiepileptic, antipsychotic, sedative and antinociceptive effects of mistletoe. Additionally, mistletoe can be used as a complementary treatment for Alzheimer's disease. The therapeutic effect of mistletoe might be a result of the synergistic interactions of various secondary metabolites, including mistletoe-specific lectins. Further studies of the chemical composition and CNS activity of mistletoe are required. The mechanisms of action, target sites, pharmacokinetics, metabolic mechanisms, adverse effects and interactions of mistletoe with other drugs must also be investigated, as well.

Sulforaphane-conjugated selenium nanoparticles: towards a synergistic anticancer effect.

Sulforaphane-modified selenium nanoparticles can be prepared in a simple aqueous-phase redox reaction through reduction of selenite with ascorbic acid. The sulforaphane molecules present in the reaction mixture adsorb on the nanoparticle surface, forming an adlayer. The resulting conjugate was examined with several physicochemical techniques, including microscopy, spectroscopy, x-ray diffraction, dynamic light scattering and zeta potential measurements. As shown in in vivo investigations on rats, the nanomaterial administered intraperitoneally is eliminated mainly in urine (and, to a lesser extent, in feces); however, it is also retained in the body. The modified nanoparticles mainly accumulate in the liver, but the basic parameters of blood and urine remain within normal limits. The sulforaphane-conjugated nanoparticles reveal considerable anticancer action, as demonstrated on several cancer cell cultures in vitro. This finding is due to the synergistic effect of elemental selenium and sulforaphane molecules assembled in one nanostructure (conjugate). On the other hand, the cytotoxic action on normal cells is relatively low. The high antitumor activity and selectivity of the conjugate with respect to diseased and healthy cells is extremely promising from the point of view of cancer treatment.

Analysis of Menaquinone-7 Content and Impurities in Oil and Non-Oil Dietary Supplements.

Rapid, global technological development has caused the food industry to use concentrated food component sources like dietary supplements ever more commonly as part of the human diet. This study analysed the menaquinone-7 (MK-7) content of dietary supplements in oil capsule and hard tablet forms. A novel method for separating and measuring geometric isomers of MK-7 in dietary supplements was developed and validated. Eleven different isomers of cis/trans- menaquinone-7 were identified. Identification of cis/trans isomers was performed by combination of HPLC, UPLC and HRMS-QTOF detection, whereas their quantities were determined by DAD detection. The content of menaquinone impurities was ascertained, including cis/trans- menaquinone-6 isomers (5.5⁻16.9 µg per tablet/capsule) and cis/trans-menaquinone-7 isomers (70.9⁻218.7 µg tablet/capsule), which were most likely formed during the chemical synthesis of the menaquinone-7. The all-trans MK-7 content was lower than the isomeric form and often lower than what the labels declared. A new method of quantification, developed and validated for menaquinones in oil capsules, provided on average 90% recovery and a limit of quantification (LOQ) of approximately 1 µg mL−1.

Identification of cis/trans isomers of menaquinone-7 in food as exemplified by dietary supplements.

For the first time, the cis/trans isomer content of menaquinone-7 in food products has been identified and marked. A novel method of marking isomers of vitamin K2MK-7 in dietary supplements was developed and validated. Five different isomers of cis/trans vitamin K2MK-7 were identified. Identification of cis/trans isomers was performed by HRMS-QTOF, whereas their quantities were determined by using CAD and DAD detectors. In the majority of cases, the content of biologically active all trans vitamin K2MK-7 was below its declared content. The content of all trans K2MK-7 was in the range between 5.5 and 49 µg in pills. In one of the studied supplements, this vitamin was not found, regardless of the claim on the product label. The content of cis/trans isomers in certain dietary supplements exceeded the content of all trans K2MK-7 by up to 3.7 times.

Nutritional value of raw pork depending on the fat type contents in pigs feed.

BACKGROUND: The research aimed to assess the nutritional value of raw pork meat obtained from pigs on the feed containing different types of used fat. METHODS: Meat was obtained from pigs fed in 4 experimental groups, from which groups A and B were given the mixture of rapeseed oil and linseed oil in the ratio of 1:2.5 or 2.3:1:0.2 (lard) as the source of fat. In group C, animals were given the mixture of rapeseed oil (1 part), fish oil (cod-liver oil; 2 parts) and lard (0.5 part), whereas group D obtained the mixture of linseed oil in the same ratio as in group B (2.5 parts), but combined with cod-liver oil (1 part). Meat collected for the research was evaluated and underwent chemical analyses in order to determine its nutritional value. The contents of moisture, protein, fat and ash were determined according to the AOAC methods. The carbohydrate content in various samples was determined by subtracting the sum of moisture, protein, ash and fat content from 100. Energy value (EV) was calculated using Atwater factors. The fatty acids content were analysed in homogenized samples using Folch method. Methyl esters of fatty acids were separated by gas chromatography on GC Trace Ultra THERMO company equipped with a RT 2560 RESTEC capillary column (100 m × 0.25 mm × 0.2 µm) with helium as a carrier gas. Result. The modification of the source of fat in pigs feed did not influence the content of protein and total minerals (ash), whereas it significantly influenced the content of fat and moisture. It has been stated that the use of different fat type in pigs feed, contributed to receiving the meat with modified profile of fatty acids. Higher fatty acids of the n-3 was observed in groups B and D (treated with the mixture of oils with a predominance of linseed oil). The fat from the meat of these groups contain less of n-6 and n-9 fatty acid, which improved the quality indicator of the fat expressed as the ratio of n-6 / n-3. Meat obtained from animals of the groups contained significantly less MUFA, and comparable content UFA and SFA. The meat from all experimental groups the ratio of polyunsaturated fatty acids (PUFA), saturated fatty acids (SFA) remained at a similar level and was 0.5:0.6. Conclusions. The modification of the source of fat in pigs feed did not influence the content of protein and total minerals (ash), whereas it significantly influenced the content of fat and moisture. A different source of fat in pigs feed did not influence the content of fat, saturated fatty acids (SFA) and UFA (unsatur. RESULTS: nd. The research aimed to assess the nutritional value of raw pork meat obtained from pigs on the feed containing different types of used fat. Material and methods. Meat was obtained from pigs fed in 4 experimental groups, from which groups A and B were given the mixture of rapeseed oil and linseed oil in the ratio of 1:2.5 or 2.3:1:0.2 (lard) as the source of fat. In group C, animals were given the mixture of rapeseed oil (1 part), fish oil (cod-liver oil; 2 parts) and lard (0.5 part), whereas group D obtained the mixture of linseed oil in the same ratio as in group B (2.5 parts), but combined with cod-liver oil (1 part). Meat collected for the research was evaluated and underwent chemical analyses in order to determine its nutritional value. The contents of moisture, protein, fat and ash were determined according to the AOAC methods. The carbohydrate content in various samples was determined by subtracting the sum of moisture, protein, ash and fat content from 100. Energy value (EV) was calculated using Atwater factors. The fatty acids content were analysed in homogenized samples using Folch method. Methyl esters of fatty acids were separated by gas chromatography on GC Trace Ultra THERMO company equipped with a RT 2560 RESTEC capillary column (100 m × 0.25 mm × 0.2 µm) with helium as a carrier gas. Result. The modification of the source of fat in pigs feed did not influence the content of protein and total minerals (ash), whereas it significantly influenced the content of fat and moisture. It has been stated that the use of different fat type in pigs feed, contributed to receiving the meat with modified profile of fatty acids. Higher fatty acids of the n-3 was observed in groups B and D (treated with the mixture of oils with a predominance of linseed oil). The fat from the meat of these groups contain less of n-6 and n-9 fatty acid, which improved the quality indicator of the fat expressed as the ratio of n-6 / n-3. Meat obtained from animals of the groups contained significantly less MUFA, and comparable content UFA and SFA. The meat from all experimental groups the ratio of polyunsaturated fatty acids (PUFA), saturated fatty acids (SFA) remained at a similar level and was 0.5:0.6. CONCLUSIONS: The modification of the source of fat in pigs feed did not influence the content of protein and total minerals (ash), whereas it significantly influenced the content of fat and moisture. A different source of fat in pigs feed did not influence the content of fat, saturated fatty acids (SFA) and UFA (unsaturated fatty acids) in fat, whereas it significantly influenced the participation of  particular fatty acids belonging to monounsaturated (MUFA) and n-3 and n-6 groups.

Chemical Stability of the Lipid Phase in Concentrated Beverage Emulsions Colored with Natural ß-Carotene.

The aim of this study was to examine the oxidation of selected plant oils in concentrated beverage emulsions colored with natural ß-carotene. Carotenoid preparations obtained from carrots were dissolved in cold-pressed linseed oil, refined canola oil, and refined palm olein. Oxidative stability of the lipids was examined with and without addition of the pigment to the oil/water (O/W) emulsion. Carotenoid/lipid hydro peroxide (LOOH) concentration was evaluated using two different methods: LOOH + Fe2+ reaction connected with a colored complex of ammonium thiocyanate determined with the help of a spectrophotometer, and LOOH determined with the help of a chemiluminometer. It was shown that oxidation rate of lipids in the O/W emulsions strongly depended on chemical composition of the lipid fraction (type of oil used). Presence of the carotenoid pigment increased the rate. Therefore, if a carotenoid-containing emulsion is to be stable, it should be based on oils of a high oxidative stability.

Determination of azaarenes in oils using the LC-APCI-MS/MS technique: new environmental toxicant in food oils.

A novel method to determine of azaarenes in refined and cold-pressed vegetable oils and animal fats is reported. The method may be used to determine eight most important acridine derivatives (benz[a]acridine, dibenz[a,i]acridine, benz[c]acridine, dibenz[a,j]acridine, 7,9-dimethylbenz[c]acridine, dibenz[a,h]acridine, dibenz[a,c]acridine, dibenz[c,h]acridine) at a high sensitivity (LOQ in the 2-25 ng kg(-1) range), high analyte recovery rates (70.7-98.7%), sufficient linearity within the studied concentration range (r > 0.97). The method is fast, simple, and needs no expensive clean-up procedures to successfully determine the analytes. Azaarene concentration in the studied oil samples ranged from 2 to 250 ng kg(-1). Benz[a]acridine and dibenz[a,j]acridine were the compounds found most commonly and at the highest concentrations. The observed concentrations most probably reflected levels of environmental contamination of raw materials used to produce the analyzed oil/fat samples.

New opportunities of the application of natural herb and spice extracts in plant oils: application of electron paramagnetic resonance in examining the oxidative stability.

The aim of this study was to establish the applicability of natural water-ethanol extracts of herbs and spices in increasing the oxidative stability of plant oils and in the production of novel food. Different concentrations (0, 100, 300, 500, and 700 ppm) of spice extracts and butylated hydroxyanisole (BHA) (100 ppm) were added to the studied oils. The antioxidant activity of spice extracts was determined with electron paramagnetic resonance (EPR) spectroscopy using 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical assay. The study showed that the extracts significantly increased the oxidative stability of the examined oils when compared to one of the strongest synthetic antioxidants--BHA. The applied simple production technology and addition of herb and spice extracts to plant oils enabled enhancement of their oxidative stability. The extracts are an alternative to the oils aromatized with an addition of fresh herbs, spices, and vegetables because it did not generate additional flavors thus enabling the maintenance of the characteristic ones. Moreover, it will increase the intake of natural substances in human diet, which are known to possess anticarcinogenic properties.

Profiles and concentrations of heterocyclic aromatic amines formed in beef during various heat treatments depend on the time of ripening and muscle type.

Heterocyclic Aromatic Amine (HAA) profiles and concentrations depended on several factors. The largest changes in the HAA profile were observed in meat ripened (chill stored) for 5-10 days. Amines whos concentration varied most prominently included: Phe-P 1, harmane, AαC, IQ, IQx, PhIP, MeAαC, and MeIQx. HAA concentrations were strongly correlated with concentrations of the above compounds. Time of storage significantly affected the HAA profile and concentration. The profile changed dynamically for storage times up to 10 days. For longer times the profile stabilized, only the HAA content increased. A novel, highly precise and accurate HAA analytical method was developed for this study. Results may help to optimize meat processing technology from the point of view of reducing concentration of HAA formed during heat treatment, including the most carcinogenic; IQ, IQx, MeIQx and PhIP amines.