Biological Properties of the Mucus and Eggs of Helix aspersa Müller as a Potential Cosmetic and Pharmaceutical Raw Material: A Preliminary Study.

In recent years, snail mucus (SM) has become popular as an active ingredient in cosmetic and pharmaceutical products. In turn, snail eggs (SEs) also seem to be a promising active compound, but the biological activities of SEs are significantly less known. Therefore, our preliminary study aimed to compare the biological activities of the SEs and SM of Helix aspersa Müller. The metabolomic analysis (LC-MS technique), determination of the antimicrobial activity (agar diffusion test, broth microdilution methods), antioxidant activity (ABTS assay), cytotoxicity assay (MTT), and proapoptotic properties (flow cytometry) of the SEs and SM were evaluated. It was found that the SEs and SM contain 8005 and 7837 compounds, respectively. The SEs showed antibacterial activity against S. aureus (MIC 12.5 mg/mL) and P. aeruginosa (MIC 3.12 mg/mL). The EC50 estimation of the antioxidant activity is 89.64 mg/mL and above 100 mg/mL for the SEs and SM, respectively. The SEs also inhibited the cell proliferation of cancer cell lines (HCT-116, MCF-7, HT-29) more strongly compared to the SM. The highest proportion of apoptotic cells in HCT-116 was observed. The reach composition of the compounds in the SEs and SM may be crucial for the creation of new cosmetic and pharmaceutical raw materials with different biological activities. However, further extended studies on the biological activities of H. aspersa-delivered materials are still necessary.

Splenic Elemental Composition of Breast Cancer-Suffering Rats Supplemented with Pomegranate Seed Oil and Bitter Melon Extract.

The aim of this study was to investigate how dietary modifications with pomegranate seed oil (PSO) and bitter melon aqueous extract (BME) affect mineral content in the spleen of rats both under normal physiological conditions and with coexisting mammary tumorigenesis. The diet of Sprague-Dawley female rats was supplemented either with PSO or with BME, or with a combination for 21 weeks. A chemical carcinogen (7,12-dimethylbenz[a]anthracene) was applied intragastrically to induce mammary tumors. In the spleen of rats, the selected elements were determined with a quadrupole mass spectrometer with inductively coupled plasma ionization (ICP-MS). ANOVA was used to evaluate differences in elemental composition among experimental groups. Multivariate statistical methods were used to discover whether some subtle dependencies exist between experimental factors and thus influence the element content. Experimental factors affected the splenic levels of macroelements, except for potassium. Both diet modification and the cancerogenic process resulted in significant changes in the content of Fe, Se, Co, Cr, Ni, Al, Sr, Pb, Cd, B, and Tl in rat spleen. Chemometric analysis revealed the greatest impact of the ongoing carcinogenic process on the mineral composition of the spleen. The obtained results may contribute to a better understanding of peripheral immune organ functioning, especially during the neoplastic process, and thus may help develop anticancer prevention and treatment strategies.

Pomegranate seed oil and bitter melon extract supplemented in diet influence the lipid profile and intensity of peroxidation in livers of SPRD rats exposed to a chemical carcinogen.

Despite promising health effects of pomegranate seed oil (PSO) and bitter melon extract (BM) used for centuries as food and traditional medicine, neither mechanism of action nor safety has been fully recognized. This study aimed to evaluate the influence of diet supplementation with PSO and BM on fatty acid, conjugated fatty acid and cholesterol content in rat' livers, since liver is crucial for lipid metabolism. Oxidation indicators (malondialdehyde, oxysterols and tocopherols) were also determined. Lipid profiles did not reveal the presence of punicic acid, while other conjugated dienes and trienes, including rumenic acid, were determined. Both supplementation and exposition to carcinogen significantly increased cholesterol and reduced selected oxysterols levels, simultaneously increasing malondialdehyde content in animals suffering from cancer. Impact of PSO and BM on oxidative status varied depending on carcinogen exposure and coexisting neoplastic process, which is important, due to the growing interest in their use in prevention and therapy of various diseases, including cancer.

Lipidomic Profile and Enzymes Activity in Hepatic Microsomes of Rats in Physiological and Pathological Conditions.

Among the risk factors affecting the development of cancer, nutritional factors occupy a significant place. Pomegranate seed oil (PSO) and bitter melon extract (BME), used for ages in folk medicine, are nowadays used in the prevention of many diseases and as ingredients of dietary supplements. Despite numerous publications on these raw materials or their active substances, their mechanism of action in various pathological states has not been recognized yet, nor has the safety of their simultaneous use been evaluated. The study aimed to assess how dietary supplementation with either PSO, with BME, or both, affects fatty acids' profiles and their metabolism in hepatic microsomes, as well as the activity of selected microsomal enzymes (COX-2 and CYP1B1). Experimental animals (Sprague-Dawley rats) were divided into eight parallel experimental groups, differing in applied dietary modifications (control, PSO, BME and both PSO and BME) and introduction of chemical carcinogen-7,12-dimethylbenz[a]nthracene. Obtained results indicated the pronounced effect of the cancerous process on lipid metabolism and demonstrated the antagonistic effect of applied dietary supplements on the content of individual fatty acids and the activity of CYP1B1 and COX-2. The applied broad analytical approach and chemometric data analysis confirmed that raw materials, for which potential cancer prevention has been previously demonstrated, may differ in effects depending on the coexisting pathological state.

Cancer Influences the Elemental Composition of the Myocardium More Strongly than Conjugated Linoleic Acids-Chemometric Approach to Cardio-Oncological Studies.

The aim of the study was to verify in a cardio-oncological model experiment if conjugated linoleic acids (CLA) fed to rats with mammary tumors affect the content of selected macro- and microelements in their myocardium. The diet of Sprague-Dawley females was supplemented either with CLA isomers or with safflower oil. In hearts of rats suffering from breast cancer, selected elements were analyzed with a quadrupole mass spectrometer with inductively coupled plasma ionization (ICP-MS). In order to better understand the data trends, cluster analysis, principal component analysis and linear discriminant analysis were applied. Mammary tumors influenced macro- and microelements content in the myocardium to a greater extent than applied diet supplementation. Significant influences of diet (p = 0.0192), mammary tumors (p = 0.0200) and interactions of both factors (p = 0.0151) were documented in terms of Fe content. CLA significantly decreased the contents of Cu and Mn (p = 0.0158 and p = 0.0265, respectively). The level of Ni was significantly higher (p = 0.0073), which was more pronounced in groups supplemented with CLA. The obtained results confirmed antioxidant properties of CLA and the relationship with Se deposition. Chemometric techniques distinctly showed that the coexisting pathological process induced differences to the greater extent than diet supplementation in the elemental content in the myocardium, which may impinge on cardiac tissue's susceptibility to injuries.

Giblets and abdominal fat of pomegranate seed oil fed chickens as a source of bioactive fatty acids.

The aim of present study was to determine whether chickens' (broilers Ross 308, n = 180, sex ratio 1:1) diet modification with different doses of grape or pomegranate seed oil will favourable change fatty acids and cholesterol content in selected giblets (liver and heart) or wastes (adipose tissue). It was also verify whether generated changes would make the giblets and wastes more valuable as dietary components or by-products for food industry. From 22 to 42 day of life, five diets were administered to chickens. Control grower diet (CON) contained 5% of soy oil, whereas in the experimental grower diets part of soy oil (1.5% or 2%) was replaced with specific amount of grape or pomegranate seed oil (GRAP 1.5; GRAP 2.0; POM 1.5; POM 2.0 respectively). Fatty acids and cholesterol content were determined with gas chromatography with flame-ionization detection. Pomegranate seed oil improved fatty acids profile more favourably than grape seed oil, which makes it a valuable additive in chickens' feeding. Abdominal fat of pomegranate seed oil supplemented chickens appeared to be the richest sources of rumenic acid and n3 polyunsaturated fatty acids, which allows to suggest its use in manufacturing of meat products to obtain foodstuffs rich in those essential nutrients. In principal component analysis (PCA), two principal components: PC1 and PC2, which were enough to explain 29.91% of variance of initial variables, allowed to a good separation of chickens fed with both doses of pomegranate seed oil from animals from control and grape seed oil fed groups. Because poultry addresses all nutritional, institutional and consumer requirements, enrichment of giblets in rumenic acid by pomegranate seed oil incorporation into chickens' diet may provide a valuable dietary source of bioactive fatty acids for consumers, especially of low-income countries.

Chemometric Analysis of Fatty Acids Profile of Ripening Chesses.

The number of different types of cheese worldwide exceeds 4000 and dairy fat, composed of about 400 fatty acids (FA), is one of the most complex dietary fats. Cheeses are valuable sources of different bioactive FA, i.e., conjugated FA (CFA). The aim of present study was to determine FA profile of commercially available ripening cheeses, with the special emphasis on CFA profile. Multivariate analyses (cluster analysis (CA), principal component Analysis (PCA), and linear discriminant analysis (LDA)) of chromatographic data have been proposed as an objective approach for evaluation and data interpretation. CA enabled the differentiation of ripening cheeses from fresh cheeses and processed cheeses. PCA allowed to differentiate some types of ripening cheese whereas proposed LDA model, based on 22 analyzed FA, enabled assessing cheeses type with average predictive sensitivities of 86.5%. Results of present study clearly demonstrated that FA and CFA content may not only contribute to overall nutritional characteristics of cheese but also, when coupled with chemometric techniques, may be used as chemical biomarkers for assessing the origin and/or the type of ripening cheeses and the confirmation of their authenticity, which is of utmost importance for consumers.

Mammary cancer risk and serum lipid profile of rats supplemented with pomegranate seed oil and bitter melon extract.

The objective of the present study was to determine the influence of dietary supplementation with pomegranate seed oil (PSO) and/or an aqueous extract of dried bitter melon fruits (BME) on breast cancer risk and fatty acid profile in serum of female rats with chemical carcinogen-inflicted mammary tumours. Sprague-Dawley rats (n = 96) were fed control diet or experimental diets supplemented with 0.15 ml PSO/day, BME or jointly PSO and BME. After 21 weeks mammary tumours were subjected to histopathological examination and in serum fatty acids, 8-isoprostaglandin F2α content and indices of desaturases activity were analysed. Supplementation of the diet with PSO and BME did not inhibit the breast cancer formation. Conjugated linolenic acids (CLnA), present in PSO, were converted into cis-9, trans-11 conjugated linoleic acid (CLA), however, its content was lower in groups treated with a carcinogen. A similar tendency was observed for the content of SFA, MUFA, PUFA, 8-iso PGF2α and the activity of Δ6-desaturase. Enhanced pro-carcinogenic effect of 7,12-dimethylbenz[a]anthracene (DMBA), caused by applied supplements, may be a result of their influence on DMBA metabolism.

Pomegranate seed oil influences the fatty acids profile and reduces the activity of desaturases in livers of Sprague-Dawley rats.

The aim of our study was to compare the influence of diet supplementation with pomegranate seed oil - as conjugated linolenic acids (CLnA) source, or conjugated linoleic acids (CLA) and to examine the mechanism of their activity. The content of fatty acids, levels of biomarkers of lipids' oxidation and the activity of key enzymes catalyzing lipids metabolism were measured. Obtained results revealed that conjugated fatty acids significantly decrease the activity of Δ5-desaturase (p=0.0001) and Δ6-desaturase (p=0.0008) and pomegranate seed oil reduces their activity in the most potent way. We confirmed that diet supplementation with pomegranate seed oil - a rich source of punicic acid leads to the increase of cis-9, trans-11 CLA content in livers (p=0.0003). Lack of side effects and beneficial influence on desaturases activity and fatty acids profile claim pomegranate seed oil to become interesting alternative for CLA as functional food.

INFLUENCE OF POMEGRANATE SEED OIL AND GRAPE SEED OIL ON CHOLESTEROL CONTENT AND FATTY ACIDS PROFILE IN LIVERS OF CHICKENS.

Oils are important food ingredients, mainly as a source of unsaturated fatty acids. The offer of novel edible oils from herbs, spices and fruit seeds has grown and many of them are used as functional food and dietary supplements but also as feed additives in animal feeding. Poultry meat is recommended.in properly balanced diet and its consumption in Poland has been growing. The objective of present study was to verify if the supplementation of chickens' diet with grape seed oil or pomegranate seed oil influences cholesterol content and fatty acids (FA) profile in their livers. Ross 308 chickens (n = 24) were fed with fodder enriched with grape seed oil (G group) or pomegranate seed oil (P group). Diet of control group (C group) was based on soybean oil. FA analysis in livers as well as cholesterol content was made with gas chromatography. We observed significant increase in fat content when part of soybean oil was replaced by grape seed oil (p = 0.0002). Its highest amount was detected in G group (4.44 ± 1.53%) whereas the lowest in C group (1.73 ± 0.53%). Applied supplementation did not change total cholesterol content. Its content ranged from 233.0 ± 12.2 mg/100 g in G group to 234.6 ± 29.7 mg1100 g in C group. However, chickens' diet modification with grape seed oil and pomegranate seed oil influenced the FA profile in livers. We detected the presence of punicic acid (cis-9, trans-11, cis-13 C18:3, PA) in livers of chicken fed with pomegranate seed oil. Pomegranate seed oil is one of natural sources of conjugated linolenic acids (CLnA), which predominate in this oil (PA >70% of all FA). However, in livers PA constituted only 0.90 ± 0.10% of all fatty acids. Furthermore, we detected substantial amounts of rumenic acid (cis-9, trans-11 C18:2, RA) - the major isomer of conjugated linoleic acids (CLA). Its natural sources in diet are meat and milk of ruminants, but incorporation of pomegranate seed oil into chickens' diet caused a significant increase of its share in fatty acids pool in their livers (3.73 ? 0.79% in P group in relation to 0.08 ± 0.03% in G group and 0.02 ± 0.00% in C group, p < 0.0001). It proves that PA is effectively converted into RA in chickens organisms. Pomegranate seed oil seems to be an interesting feed additive in chicken feeding which can improve FA profile of poultry meat.

Effects of replacing extracted soybean meal with rapeseed cake in corn grass silage-based diet for dairy cows.

The aim of the study was to assess the effects of partial replacement of soybean meal with a protein-equivalent amount of rapeseed cake in the diet on milking parameters and fatty acid (FA) composition of milk in dairy cows. Two groups of Holstein-Friesian cows, 8 each, consisting of randomised blocks were studied: a control group (C) was given a traditional high-protein supplement (extracted soybean meal) and the experimental group (E), had part of extracted soybean meal replaced with rapeseed cake. Dry matter intake and milk yield in both groups were not affected by the diet but milk fat percentage and yield were decreased in both groups. Rapeseed cake had no effect on milk acidity or on protein (including casein) and lactose contents. A lower concentration of urea in milk in E group indicated a proper ratio of protein to energy in the fodder. Health condition of mammary gland and indicators of metabolic profile were not affected by rapeseed cake supplementation. In E group, the share of atherogenic saturated fatty acids (FA) was reduced after 11 weeks: palmitic, by 26% and myristic, by 22%; moreover, as compared with control cows, the content of monounsaturated FA in milk increased by 44% after 3 weeks and by 68% after 11 weeks, t-18:1 and c-9 t-11 isomer of CLA increased about 2.5-fold after 11 weeks. In E group, the atherogenic index (AI) was significantly (P < 0.001) lower than in C (by 54% on average) and the decrease with time was considerable (by 29%, P < 0.001). Contents of odd- and branched- chain FA in milk were not significantly affected thus reflecting proper rumen function. Partial replacement of soybean meal with rapeseed cake in the diet of cows may improve both milking indices and FA profile of milk.

[The importance of γ-linolenic acid in the prevention and treatment]. / Znaczenie kwasu γ-linolenowego w profilaktyce i terapii.

The etiology of diet-related disorders is closely associated with dietary factors. A special role is attributed to intake of fat and fatty acid profile, both quantitative and qualitative. For prevention and treatment of the abovementioned diseases a proper supply of unsaturated fatty acids plays a significant role, because of their particular importance to health. γ-Linolenic acid (GLA), with three double bonds in the carbon chain, also known as all-cis 6,9,12-octadecatrienoic acid, belongs to the n-6 family of fatty acids. It plays biologically important functions in the human body, such as being a substrate for eicosanoids synthesis, involvement in the transport and oxidation of cholesterol, and being one of the components of lipid membrane. Its inadequate dietary intake or impaired formation is the cause of many inflammatory and degenerative diseases. A rich source of this fatty acid is vegetable oils, until recently used mainly in folk medicine. Nowadays, studies conducted both in animal models and in humans suggest its health-promoting properties in the prevention and treatment of atopic dermatitis, cardiovascular diseases, diabetes, cancers and rheumatoid arthritis.

Cholesterol and Its Oxidation Derivatives Content in Market Dairy Products.

Cholesterol oxidation products (COPs) are contaminants of food of animal origin. Increased levels of these compounds in the human body are associated with an increased risk of many non-communicable diseases. Dairy products are mentioned among the main sources of these compounds in the diet. The objective of this study was to evaluate the contents of cholesterol and its oxidized derivatives in eleven groups of dairy products, willingly consumed in European countries. The levels of COPs were determined by applying the GC-TOF/MS method. In the tested products, cholesterol and its oxidation derivatives, such as 7-ketocholesterol, 7α-hydroxycholesterol, 7ß-hydroxycholesterol, 5,6ß-epoxycholesterol and 5,6α-epoxycholesterol, were determined. The studied dairy products differed in their contents and profiles of oxysterols. The highest contents of COPs were found in cheese with internal mold (13.8 ± 2.5 mg kg-1) and Cheddar (11.7 ± 3.5 mg kg-1), while the lowest levels were detected in yoghurt (0.94 ± 0.30 mg kg-1) and kefir (0.57 ± 0.11 mg kg-1). 7-ketocholesterol and 5,6ß-epoxycholesterol were the dominant oxysterols. The ratio of oxidized derivatives to total cholesterol was on average 1.7%. Our results confirmed that dairy products are an important dietary source of COPs. Their levels should be monitored in dairy products to provide the best health quality.

Exploring the Influence of the Selected Conjugated Fatty Acids Isomers and Cancerous Process on the Fatty Acids Profile of Spleen.

The spleen, traditionally associated with blood filtration and immune surveillance, has recently been recognized for its role in systemic lipid metabolism and potential influence on cancer development and progression. This study investigates effects of dietary supplements, specifically conjugated linolenic acids from pomegranate seed oil and bitter melon extract, on the fatty acid (FA) composition of the spleen in the context of cancerous processes. Advanced methods, including gas chromatography-mass spectrometry and silver ion-impregnated high-performance liquid chromatography, were employed to analyze the spleen's FA profile. Our research uncovered that dietary supplementation leads to alterations in the spleen's FA profile, especially under the carcinogenic influence of 7,12-dimethylbenz[a]anthracene. These changes did not align with a simple protective or anti-carcinogenic pattern, as previously suggested in in vitro studies. We observed shifts in conjugated FA isomer concentrations and variations in desaturase activities, suggesting disrupted lipid metabolism in cancerous conditions. The findings underscore the spleen's vital role in lipid metabolism within the body's systemic health framework, highlighting the complexity of dietary supplements' impact on FA profiles in the spleen and their potential implications in cancer progression and treatment. This study adds valuable insight into the complex interplay between diet, disease, and metabolic regulation, particularly in cancerous environments.

Organic and Inorganic Selenium Compounds Affected Lipidomic Profile of Spleen of Lambs Fed with Diets Enriched in Carnosic Acid and Fish Oil.

The purpose of our study was to investigate the effect of 0.35 mg Se/kg basal diet (BD) (Se as sodium selenate (Se6) and yeast rich in seleno-methionine (SeYe)) and 0.1% carnosic acid (CA) supplementation to the diet containing 1% fish oil (F-O) and 2% rapeseed oil (R-O) on the contents of fatty acids (FA), malondialdehyde (MDA), tocopherols (Ts), and total cholesterol (TCh) in lambs' spleens. A total of 24 male lambs (4 groups per 6 animals) have been fed: the control diet-the basal diet (BD) enriched in F-O and R-O; the CA diet-BD enriched in F-O, R-O, and CA; the SeYeCA diet-BD enriched in F-O, R-O, CA, and SeYe; the Se6CA diet-BD enriched in F-O, R-O, CA, and Se6. Dietary modifications affected the profiles of saturated (SFA), monounsaturated (MUFA), and polyunsaturated (PUFA) fatty acids in spleens. The SeYeCA and Se6CA diets increased the docosapentaenoic acid preference in Δ4-desaturase; hence, a higher content of docosahexaenoic acid was found in the spleens of SeYe- or Se6-treated lambs than in spleens of animals receiving the CA and control diets. The SeYeCA and Se6CA diets increased the concentration ratio of n-3long-chain PUFA (n-3LPUFA) to FA (n-3LPUFA/FA) in spleens compared to the control and CA diets. The content of n-3PUFA was higher in the spleens of Se6 treated lambs than in spleens of animals receiving the SeYeCA, CA, and control diets. The Se6CA diet increased the content of c9t11CLA in the spleen compared to the control, CA, and SeYeCA diets. Experimental diets reduced the level of atherogenic FA, the content ratios of n-6PUFA/n-3PUFA and n-6LPUFA/n-3LPUFA, and improved the content ratio of MUFA/FA and the value of the hypocholesterolemic/hypercholesterolemic FA ratio in the spleen in comparison with the control diet. The experimental diets supplemented with SeYe or Se6 increased levels of TCh and Ts in spleens in comparison with the CA and control CA diets. The present studies documented that Se6, SeYe, and CA influenced the metabolism of FA, Ts, and cholesterol in spleens.

Selected Exogenous (Occupational and Environmental) Risk Factors for Cardiovascular Diseases in Military and Aviation.

Cardiovascular diseases are a group of disorders of heart and blood vessels which are the leading cause of death globally. They are also a very important cause of military unsuitability especially among military pilots. Some of the risk factors cannot be modified, but a large group of them can be modified by introducing healthy habits or behaviors, such as regular physical activity, quitting of tobacco smoking, cessation of harmful alcohol consumption, or stress avoidance. Specific occupational and environmental factors, such as acceleration, pressure changes, hypoxia, thermal stress, noise, vibration, prolonged sedentary posture, difficulties in moving the limbs, shifts, work shift irregularities, circadian dysrhythmia, variations in the microclimate, motion sickness, radiation, toxicological threats, occupational stress, emotional tension, highly demanding tasks especially during take-off and landing, predominance of intellectual over physical activity, and forced operation speed against time shortage are considered as characteristic for military personnel, especially military aircrews. They are of special concern as some of them are considered as potential CVD risk factors. The aim of this study was to discuss the influence of selected occupational and environmental factors (noise, altitude, hypoxia, acceleration forces, tobacco smoking, oral health, and stress) regarding their importance for CVD risk in military professionals and military aviators. The performed revision confirmed that they are exposed to certain characteristic conditions, which may influence the CVD risk but the number of solid scientific data regarding this subject are limited. There is a great need to perform complex studies on environmental and occupational risk factors for CVDs in military personnel and military aviators as well as how to minimize their influence to prolong the state of health and military suitability of this professional groups.

Fatty acid profile of the milk of cows reared in the mountain region of Poland.

An appropriate composition of milk fatty acids (FA) improves the nutritional value of milk and milk products, and improves milk processing. Polish dairy farms in the mountainous region are rather small, animal nutrition there is based on locally produced forages and this, together with the transitional climate zone brings about seasonal changes in FA composition of milk. The aim of the study was to evaluate the composition of FA in bovine milk fat in relation to fat intake in forages and their FA profiles. The study involved 5 herds reared in low-input mountain farms located at an altitude of 670-780 m above sea level (Beskid Mountains). The cows were fed forages produced locally. FAs in forages and milk samples were subjected to gas chromatography. Highest fat intake observed in grazing season (4·2-4·7%) and high amounts of polyunsaturated FA in forages from that period (51·8-64·1 g/100 g FA) resulted in a markedly high content of valuable FAs: t-11 C18:1 (3·22 g/100 g FA), c-9, t-11 C18:2 (CLA; 1·20 g/100 g FA) in milk. Lower fat intake of forages containing high amount of SFA (32·42-38·83 g/100 g FA) in the indoor period resulted in changes in milk composition. The content of total short-chain saturated FA (SCFA) was highest in winter and early spring samples (14·10 and 13·44 g/100 g FA, respectively), like the amounts of myristic C14:0 and palmitic C16:0 acids (11·80 and 37·92 g/100 g FA). Total odd- and branched-chain fatty acids (OBCFA; 6·58 g/100 g FA) content was highest at the beginning of the grazing period. Fresh grass consumed by cows promoted the activity of Δ(9)-desaturase in mammary gland as evidenced by higher C14:1 : C14:0 (0·054) and C16:1 : C16:0 (0·026) ratios in grazing than in the indoor periods.

New insights into NAFLD based on preclinical MRI studies.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is considered to be the leading cause of liver pathologies worldwide. Accurate diagnosis and staging of NAFLD is of utmost prognostic importance. Herein, we propose the application of multiparametric MRI in whole-body fat imaging that may serve in obesity and NAFLD diagnosis as we established based on an experimental preclinical model of high-calorie diet-induced NAFLD rats. METHODS AND RESULTS: Sprague Dawley male rats randomly divided into control and high-fat diet (HFD) groups to develop NAFLD were used in the experiments. After 12 weeks of the feeding the animals were subjected to MRI modalities based on the Dixon mode and DWI following T2-weighted imaging. Moreover, serum TAG, liver histopathological examination and liver fatty acids analysis (GC-MS) were also performed. The qualitative analysis of DWI images revelated the decrease of signals in the liver of rats subjected to HFD. The statistical analysis of signals from the water- and fat-separated voxels on Dixon images also showed the increase of the fat tissue along with the decrease of water molecules in the liver parenchyma of obese animals. The quantitative analysis of Dixon images allowed to estimate the statistically significant changes of whole-body fat profiles in both normal and obese rats. Histological analysis of the liver tissues, serum TAG and fatty acids profile in the livers confirmed the changes in the fat profile as demonstrated in MRI studies. CONCLUSIONS: The MRI-based modalities such as DWI and Dixon method provide both qualitative and quantitative data on the whole-body fat distribution and adipose tissues in the liver parenchyma of obese rats. The results show that MRI is a promising and reliable method and has potential to be used as a non-invasive translational biomarker in NAFLD.

Dietary carnosic acid and seleno-compounds change concentrations of fatty acids, cholesterol, tocopherols and malondialdehyde in fat and heart of lambs.

The aim of the current study was to evaluate the impact of carnosic acid (CA), selenised yeast (YSe) and selenate (VISe) supplemented to diets, including fish oil (FO) and rapeseed oil (RO), on the content of fatty acids, total cholesterol (TCh), tocopherols and malondialdehyde in the fat located between the thigh muscles and the heart in lambs. Twenty-four male Corriedale lambs were divided into 4 groups of 6 animals. Animals were fed a diet with FO and RO (the control diet) or experimental diets containing RO, FO and CA with/without Se (as YSe or VISe). The experimental diets without/with YSe or VISe changed concentrations of fatty acids in the fat and heart compared to the control. All experimental diets increased the levels of c11c14C20:2, c5c8c11c14C20:4, c5c8c11c14c17C20:5 and the sums of long-chain polyunsaturated fatty acids (LPUFA) and conjugated linoleic acid isomers in the fat compared to the control. The experimental diet containing YSe or VISe increased the content of Se, TCh, c11c14C20:2, c8c11c14C20:3, c5c8c11c14C20:4, c5c8c11c14c17C20:5, c7c10c13c16c19C22:5, c4c7c10c13c16c19C22:6 and the concentration sum of n-3LPUFA, n-6LPUFA and tocopherols in the heart in comparison with the control diet and the diet containing only CA. Experimental diets reduced the concentration of malondialdehyde in the fat and heart in comparison with the control diet. Our dietary intervention has great potential for future practical and commercial implementations.

The Improved Method for Determination of Orotic Acid in Milk by Ultra-Fast Liquid Chromatography with Optimized Photodiode Array Detection.

Ultra-fast liquid chromatography (UFLC) with a photodiode array detector (DAD) for simple and rapid determination of orotic acid (OAc) in milk of sheep and cows is described. Milk samples are treated with acetonitrile (1:1, v/v) and then centrifuged at 4 °C. To 1 mL of the obtained supernatant 9 mL of ultrapure water was added. Subsequently, 0.5-6 µL of the resulting solution was injected into the UFLC-DAD system. Separation and quantification of OAc in milk samples was achieved using two Kinetex C18 columns (1.7 µm, 150 mm × 2.1 mm, i.d., 100 Å; Phenomenex) fitted with a pre-column of 4 mm × 2 mm, i.d. (Phenomenex) containing C18 packing material. All separations were performed at a column temperature of 35 °C while the ambient temperature was 21-24 °C. Satisfactory separation of OAc from endogenous species of milk can be achieved using the binary gradient elution program and UV detection at wavelengths 278 nm. Our original procedure resulted in suitable separation and quantification of OAc in milk samples; OAc eluted at 6.44 ± 0.03 min. The total run time of OAc analysis (including re-equilibration) was 27 min. As expected, the OAc peak was absent from the blank when the proposed gradient elution program and UV detection at 278 nm was used. The average recoveries of OAc standards added to milk samples were satisfactory (96.7-105.3%). The low inter-and intra-assay coefficient of variation derived from the measurements of OAc in cow and ovine milk samples (i.e., 0.784%, 1.283% and 0.710%, 1.221%, respectively) and in O-Ac standards (i.e., 0.377% and 0.294%, respectively), as well as high recoveries of OAc added to ovine and cows' milk (~100%) and the low detection (0.04 ng) and quantification (0.12 ng) limits point to satisfactory accuracy, precision and sensitivity of the reported method. OAc concentrations in ovine milk samples were within the range from 25 to 36 mg/L, while OAc levels in cows' milk samples was found in the range of 32-36 mg/L. Our original procedure is suitable for routine quantification of OAc in milk of ewes and cows.