Effect on N-acetil-L-cysterine in vitro on proliferation and differentiation of children deciduous teeth dental pulp stem cells

Zubna pulpa vodi poreklo od embrionalnog ektomezenhima i potencijalno je vaţan izvor mezenhimalnih matičnih ćelija (MMĆ) za regeneraciju svih tkiva kraniofacijalne regije koja su, takođe, poreklom od ektomezenhima. N-acetil-L-cistein (NAC) je antioksidant koji moţe da ima uticaj na terapijsku primenu MMĆ. Cilj ovog rada je bio da utvrdi da li ćelije izolovane iz zubne pulpe mlečnih zuba dece i ekspandirane in vitro, imaju karakteristike MMĆ i da ispita dejstvo NAC-a na njihovu proliferaciju i diferencijaciju, kao i da se utvrdi da li NAC utiče na aktivnost enzima antioksidativne zaštite, oksidativno oštećenje različitih ćelijskih struktura i metabolizam glukoze. Metodom tkivnog eksplanta, iz šest pulpi mlečnih zuba dece, je dobijena početna populacija adheretnih ćelija. Za procenu broja CFU-F (eng. Colony Forming Unit – Fibroblast), ćelije su zasejavane u maloj gustini. Proliferativni potencijal i vreme udvajanja broja ćelija u kulturi je praćeno brojanjem ćelija posle tripsinizacije subkonfluentnih kultura, svaka 4 dana, tokom 40 dana. Protočna citometrija je korišćena za imunofenotipizaciju ex vivo umnoţenih ćelija, određivanje aktivnosti aldehid- dehidrogenaze (ALDH), brzinu ulaska ćelija u sintetsku (S) fazu ćelijskog ciklusa, određivanje broja ćelijskih deoba i detekciju apoptoze. Aktivnost -galaktozidaze (SA-β-Gal), određivana je korišćenjem citohemije. Diferencijacija ekspandiranih ćelija u smeru osteogeneze, hondrogeneze i adipogeneze izvedena je korišćenjem komercijalnih medijuma. Promene na ćelijama tokom osteogene diferencijacije su praćene preko aktivnosti alkalne fosfataze spektrofotometrijski, deponovanja kalcijuma u ekstracelularni matriks (bojenje alizarin crvenim) i pojave osteokalcina (imunocitohemija). Hondrogena diferencijacija u peletama je praćena određivanjem kolagena tip I (in situ hibridizacija), kolagena tip 2 (imunohistohemija) i određivanjem koncentracije glikozaminoglikana (spektrofotometrija). Adipogena diferencijacija je ispitana vizuelizacijom masnih kapljica (Oil red O bojenje). Aktivnost katalaze i superoksid-dismutaze (SOD) u ćelijskom lizatu je određena spektrofotometrijski. Koncentracija malondialdehida (MDA) je određena reakcijom sa tiobarbiturnom kiselinom (TBA). Karbonilni derivati proteina određivani su reakcijom sa 2,4 -dinitrofenilhidrazinom. Posle ekstrakcije ukupnih lipida i metilovanja masnih kiselina, metil-estri masnih kiselina su razdvajani gasno-tečnom hromatografijom (GLC), a koncentracija zasićenih (SFA), mononezasićenih (MUFA) i i polinezasićenih masnih kiselina (PUFA) je izraţena procentualno u odnosu na ukupne masne kiseline. Izoenzimski oblici laktat-dehidrogenaze (LDH1, LDH2, LDH3, LDH4 i LDH5) su određivani vertikalnom elektroforezom. Sva navedena merenja su izvedena bez i sa različitim koncentracijama NAC-a (0,1 mM, 1 mM i 2 mM). Naši rezultati su pokazali da je iz zubne pulpe mlečnih zuba dece dobijena populacija ćelija koja formira značajan broj CFU-F (4% izolovanih ćelija) i kontinuirano proliferiše tokom 40 dana, bez opadanja vremena potrebnog za udvajanje njihovog broja u kulturi. Ipak, dobijena populacija ćelija je bila heterogena po brzini deoba. Oko 12% ćelija je posle tri dana kultivacije bilo podeljeno manje od četiri puta, dok se ostatak ćelija podelio više od četiri puta. Vijabilitet ćelija je bio u proseku oko 95%, a svega 3% ćelija je bilo pozitivno na β-galaktozidazu. Tokom čitavog vremena kultivacije kapacitet za diferencijaciju u osteoblaste, adipocite i hondrocite se nije menjao. Gotovo sve ćelije su u pasaţi četiri eksprimirale CD44, CD73, kolagen tipa I, CD29, CD90 i osteonektin. Deo ćelija je eksprimirao STRO-1, CD146 i CD106. Marker matičnih ćelija hematopoeze nije detektovan u našem sistemu kultivacije. Opseg u kome se kretao procenat ćelija koje pokazuju povišenu aktivnost ALDH je iznosio od 4% do 24% i bio je u negativnoj korelaciji sa vremenom udvajanja ćelija u kulturi...Dental pulp originates from the embryonic ectomesenchyme and represents potentially important source of mesenchymal stem cells (MSCs) with the ability to regenerate all tissues of the craniofacial region, originating from the ectomesenchyme, too. N-acetyl-L-cysteine (NAC) is an antioxidant that may have an impact on the therapeutic use of MSCs. The aim of this study was to determine whether the cells isolated from the dental pulp of children deciduous teeth of children and expanded in vitro, have the characteristics of MSC, to examine the effect of NAC on their proliferation and differentiation and to determine whether NAC influences the metabolism of glucose, the activity of antioxidant enzymes and whether it reduces oxidative damage of various cellular macromolecules. The initial cell population was obtained from six pulps of deciduous teeth using ex vivo tissue explants method. The number of colonies (Colony Forming Unit – Fibroblast; CFU-F) was determined by seeding the low density cell culture. Proliferative potential and population doubling time of the cells in culture were followed by counting the cells after tripsinization of subconfluent cultures, every 4 days, respectively, during 40 days of cultivation. Flow cytometry was used for cell immunophenotypisation, aldehyde dehydrogenase (ALDH) activity determination, number of cells in different phases of cell cycle, number of cell divisions and percentage of cells in apoptosis. Activity of β-galactosidase (SA-β-Gal), was determined using cytochemistry. Osteogenesis, chondrogenesis and adipogenesis were induced using complete commercial mediums. Osteogenic differentiation was monitored via alkaline phosphatase activity (spectrophotometry), deposition of calcium in the extracellular matrix (Alizarin red staining) and the appearance of osteocalcin (immunocytochemistry). Chondrogenic differentiation was followed by measuring collagen type I (in situ hybridization), collagen type 2 (immunohistochemistry) and the determination of the concentration of glycosaminoglycans (spectrophotometry). Adipogenic differentiation was examined by visualization of fat droplets (Oil Red O staining). Activity of catalase and superoxide dismutase (SOD) in cell lysates was determined spectrophotometrically. Malondialdehyde (MDA) was determined by reaction with thiobarbituric acid (TBA). Carbonyl derivatives of proteins were determined by reaction with 2,4-dinitrophenylhydrazine. After total lipid extraction and methylation of fatty acids, fatty acid methyl esters were separated by gas-liquid chromatography, and the concentration of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids was expressed as percentage of total fatty acids detected in the sample. Isoenzyme forms of lactate dehydrogenase (LDH1, LDH2, LDH3, LDH4 and LDH5) were determined by vertical electrophoresis. All the above measurements were made with and without different concentrations of NAC (0,1 mM, 1 mM and 2 mM). Our results showed that the dental pulp of deciduous teeth of children had population of cells that formed a significant number of CFU-F (4% of the isolated cells) and continually proliferated for 40 days without decrease in the population doubling time. However, the resulting cell population was heterogeneous in terms of the division velocity. After three days of cultivation, around 12% of the cells were divided less than four times, while the remaining cells divided more than four times. Viability of cells was in average 95%, and only 3% of the cells were positive for β-galactosidase. During the entire time of the cultivation, expanded cells retained the ability to differentiate into osteoblasts, adipocytes and chondrocytes. Almost all cells expressed CD44, CD73, collagen type I, CD29, CD90 and osteonectin. Part of the cells expressed STRO-1, CD146 and CD106. Marker of hematopoietic stem cells have not been detected. Elevated ALDH activity ranged from 4% to 24% and was negatively correlated with the population doubling time..

Uticaj N-acetil-L-cisteina in vitro na proliferaciju i diferencijaciju matičnih ćelija zubne pulpe mlečnih zuba dece

Dental pulp originates from the embryonic ectomesenchyme and represents potentially important source of mesenchymal stem cells (MSCs) with the ability to regenerate all tissues of the craniofacial region, originating from the ectomesenchyme, too. N-acetyl-L-cysteine (NAC) is an antioxidant that may have an impact on the therapeutic use of MSCs. The aim of this study was to determine whether the cells isolated from the dental pulp of children deciduous teeth of children and expanded in vitro, have the characteristics of MSC, to examine the effect of NAC on their proliferation and differentiation and to determine whether NAC influences the metabolism of glucose, the activity of antioxidant enzymes and whether it reduces oxidative damage of various cellular macromolecules. The initial cell population was obtained from six pulps of deciduous teeth using ex vivo tissue explants method. The number of colonies (Colony Forming Unit – Fibroblast; CFU-F) was determined by seeding the low density cell culture. Proliferative potential and population doubling time of the cells in culture were followed by counting the cells after tripsinization of subconfluent cultures, every 4 days, respectively, during 40 days of cultivation. Flow cytometry was used for cell immunophenotypisation, aldehyde dehydrogenase (ALDH) activity determination, number of cells in different phases of cell cycle, number of cell divisions and percentage of cells in apoptosis. Activity of β-galactosidase (SA-β-Gal), was determined using cytochemistry. Osteogenesis, chondrogenesis and adipogenesis were induced using complete commercial mediums. Osteogenic differentiation was monitored via alkaline phosphatase activity (spectrophotometry), deposition of calcium in the extracellular matrix (Alizarin red staining) and the appearance of osteocalcin (immunocytochemistry). Chondrogenic differentiation was followed by measuring collagen type I (in situ hybridization), collagen type 2 (immunohistochemistry) and the determination of the concentration of glycosaminoglycans (spectrophotometry). Adipogenic differentiation was examined by visualization of fat droplets (Oil Red O staining). Activity of catalase and superoxide dismutase (SOD) in cell lysates was determined spectrophotometrically. Malondialdehyde (MDA) was determined by reaction with thiobarbituric acid (TBA). Carbonyl derivatives of proteins were determined by reaction with 2,4-dinitrophenylhydrazine. After total lipid extraction and methylation of fatty acids, fatty acid methyl esters were separated by gas-liquid chromatography, and the concentration of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids was expressed as percentage of total fatty acids detected in the sample. Isoenzyme forms of lactate dehydrogenase (LDH1, LDH2, LDH3, LDH4 and LDH5) were determined by vertical electrophoresis. All the above measurements were made with and without different concentrations of NAC (0,1 mM, 1 mM and 2 mM). Our results showed that the dental pulp of deciduous teeth of children had population of cells that formed a significant number of CFU-F (4% of the isolated cells) and continually proliferated for 40 days without decrease in the population doubling time. However, the resulting cell population was heterogeneous in terms of the division velocity. After three days of cultivation, around 12% of the cells were divided less than four times, while the remaining cells divided more than four times. Viability of cells was in average 95%, and only 3% of the cells were positive for β-galactosidase. During the entire time of the cultivation, expanded cells retained the ability to differentiate into osteoblasts, adipocytes and chondrocytes. Almost all cells expressed CD44, CD73, collagen type I, CD29, CD90 and osteonectin. Part of the cells expressed STRO-1, CD146 and CD106. Marker of hematopoietic stem cells have not been detected. Elevated ALDH activity ranged from 4% to 24% and was negatively correlated with the population doubling time...Zubna pulpa vodi poreklo od embrionalnog ektomezenhima i potencijalno je vaţan izvor mezenhimalnih matičnih ćelija (MMĆ) za regeneraciju svih tkiva kraniofacijalne regije koja su, takođe, poreklom od ektomezenhima. N-acetil-L-cistein (NAC) je antioksidant koji moţe da ima uticaj na terapijsku primenu MMĆ. Cilj ovog rada je bio da utvrdi da li ćelije izolovane iz zubne pulpe mlečnih zuba dece i ekspandirane in vitro, imaju karakteristike MMĆ i da ispita dejstvo NAC-a na njihovu proliferaciju i diferencijaciju, kao i da se utvrdi da li NAC utiče na aktivnost enzima antioksidativne zaštite, oksidativno oštećenje različitih ćelijskih struktura i metabolizam glukoze. Metodom tkivnog eksplanta, iz šest pulpi mlečnih zuba dece, je dobijena početna populacija adheretnih ćelija. Za procenu broja CFU-F (eng. Colony Forming Unit – Fibroblast), ćelije su zasejavane u maloj gustini. Proliferativni potencijal i vreme udvajanja broja ćelija u kulturi je praćeno brojanjem ćelija posle tripsinizacije subkonfluentnih kultura, svaka 4 dana, tokom 40 dana. Protočna citometrija je korišćena za imunofenotipizaciju ex vivo umnoţenih ćelija, određivanje aktivnosti aldehid- dehidrogenaze (ALDH), brzinu ulaska ćelija u sintetsku (S) fazu ćelijskog ciklusa, određivanje broja ćelijskih deoba i detekciju apoptoze. Aktivnost -galaktozidaze (SA-β-Gal), određivana je korišćenjem citohemije. Diferencijacija ekspandiranih ćelija u smeru osteogeneze, hondrogeneze i adipogeneze izvedena je korišćenjem komercijalnih medijuma. Promene na ćelijama tokom osteogene diferencijacije su praćene preko aktivnosti alkalne fosfataze spektrofotometrijski, deponovanja kalcijuma u ekstracelularni matriks (bojenje alizarin crvenim) i pojave osteokalcina (imunocitohemija). Hondrogena diferencijacija u peletama je praćena određivanjem kolagena tip I (in situ hibridizacija), kolagena tip 2 (imunohistohemija) i određivanjem koncentracije glikozaminoglikana (spektrofotometrija). Adipogena diferencijacija je ispitana vizuelizacijom masnih kapljica (Oil red O bojenje). Aktivnost katalaze i superoksid-dismutaze (SOD) u ćelijskom lizatu je određena spektrofotometrijski. Koncentracija malondialdehida (MDA) je određena reakcijom sa tiobarbiturnom kiselinom (TBA). Karbonilni derivati proteina određivani su reakcijom sa 2,4 -dinitrofenilhidrazinom. Posle ekstrakcije ukupnih lipida i metilovanja masnih kiselina, metil-estri masnih kiselina su razdvajani gasno-tečnom hromatografijom (GLC), a koncentracija zasićenih (SFA), mononezasićenih (MUFA) i i polinezasićenih masnih kiselina (PUFA) je izraţena procentualno u odnosu na ukupne masne kiseline. Izoenzimski oblici laktat-dehidrogenaze (LDH1, LDH2, LDH3, LDH4 i LDH5) su određivani vertikalnom elektroforezom. Sva navedena merenja su izvedena bez i sa različitim koncentracijama NAC-a (0,1 mM, 1 mM i 2 mM). Naši rezultati su pokazali da je iz zubne pulpe mlečnih zuba dece dobijena populacija ćelija koja formira značajan broj CFU-F (4% izolovanih ćelija) i kontinuirano proliferiše tokom 40 dana, bez opadanja vremena potrebnog za udvajanje njihovog broja u kulturi. Ipak, dobijena populacija ćelija je bila heterogena po brzini deoba. Oko 12% ćelija je posle tri dana kultivacije bilo podeljeno manje od četiri puta, dok se ostatak ćelija podelio više od četiri puta. Vijabilitet ćelija je bio u proseku oko 95%, a svega 3% ćelija je bilo pozitivno na β-galaktozidazu. Tokom čitavog vremena kultivacije kapacitet za diferencijaciju u osteoblaste, adipocite i hondrocite se nije menjao. Gotovo sve ćelije su u pasaţi četiri eksprimirale CD44, CD73, kolagen tipa I, CD29, CD90 i osteonektin. Deo ćelija je eksprimirao STRO-1, CD146 i CD106. Marker matičnih ćelija hematopoeze nije detektovan u našem sistemu kultivacije. Opseg u kome se kretao procenat ćelija koje pokazuju povišenu aktivnost ALDH je iznosio od 4% do 24% i bio je u negativnoj korelaciji sa vremenom udvajanja ćelija u kulturi..

Walnut supplementation after fructose-rich diet is associated with a beneficial fatty acid ratio and increased ACE2 expression in the rat heart

Increased fructose consumption has been linked with chronic inflammation and metabolic syndrome (MetS). Activation of the renin-angiotensin system (RAS) and NF-κB have been detected in MetS. Walnuts are a rich source of polyunsaturated omega-3 fatty acids (n-3 PUFA) that were suggested to exert anti-inflammatory effects related to cardio-metabolic health. We hypothesized that walnut supplementation has the capacity to revert unfavorable fructose-rich diet (FRD)-induced activation of cardiac RAS and NF-κB in male rats. Due to the lack of similar studies, we investigated the effects of walnut supplementation (6 weeks) on the expression of four RAS molecules (ACE, ACE2, AT1R, and AT2R) and NF-κB in rat heart after FRD (10% w/v, 9 weeks). In addition, we followed the changes in the n-6/n-3 PUFA ratio in the total pool of heart lipids after both treatments to elucidate the walnut effects on fatty acids in the heart. 36 animals (9 per group) participated in the experiment. FRD significantly increased the ACE protein level in the heart (p < 0.001). Walnut supplementation significantly increased the ACE2 protein level in the heart of FRD (p < 0.001). In addition, walnut supplementation showed a significant main effect on the arachidonic acid/eicosapentaenoic acid ratio (p = 0.004). Walnut supplementation significantly reduced this ratio, in comparison with both, the control group (C vs. FW, p < 0.05) and the FRD group (F vs. FW, p < 0.05). However, walnut treatment failed to revert the significant effect of fructose (p < 0.001) on the elevation of NF-κB protein level. Our results suggest a beneficial effect of walnut supplementation on ACE2 protein level and n-6/n-3 PUFA level in the heart of the animal model of MetS. Such results highlight the approach of omega-3-rich walnut supplementation in the stimulation of endogenous production of favorable molecules in the heart which could be an affordable nutritional treatment formaintenance of cardio-metabolic health

Polyunsaturated fatty acids phospholipids profiles in plasma and liver in Wistar rats of different age

The phospholipids class, fatty acids (FAs) composition and cholesterol content in membranes are basic determinants of the physical properties of membranes. They been shown to influence a wide variety of membrane dependent functions such as membrane transport, enzyme activity and receptor function. The FAs profile in tissues partly reflects not only the dietary fat intake, but also the efficiency of FAs metabolism in the body. We examined overall saturated (SFA), monounsaturated (MUFA) polyunsaturated (PUFA) fatty acids phospholipids profiles in plasma and liver in male Wistar rats of different age (n=10; 3 months and n=10; 22 mouths), as well as overall n-3, n-6 and n-6/n-3 ratio. We determined it by GC cromatography. Results showed deferent tissue specificity and age specificity also. In young rats plasma phospholipids MUFA were increased (p<0.01), while PUFA and n-6 were decreased (p<0.01)compared to aged rats.In liver phospholipids in young rats overall n-3 were increased (p<0.001), while overall N-6/n-3 ratio was ssignificantly decreased (p<0.001) compared to aged rats. Liver as a place of biosynthesis and degradation of FAs seems to have more pronounced changes in its phospholipids profiles in aging

Potential health benefits of blueberry and raspberry pomace as functional food ingredients: Dietetic intervention study on healthy women volunteers

The fruit juice industry generates pomace as a valuable by-product especially rich in polyphenols, dietary fibers, vitamins, minerals, and unsaturated fatty acids. In the cookies used in this study, 30% of the gluten-free flour was replaced with dried and ground blueberry and raspberry pomace, rich source of polyphenols, dietary fibers, linoleic and alpha-linolenic acid. In order to examine whether the addition of blueberry and raspberry pomace in cookie formulation can have beneficial effects on certain blood parameters and anthropometric measurements, the designed cookies were tested in 20 healthy, normally fed female subjects, aged 30–50 years (41.35 ± 8.58 years) over four-week dietetic intervention study. Significant changes in the composition of fatty acids serum phospholipids, decrease in LDL-cholesterol level (20.16%), increase in adiponectin level (25.52%) and decrease in ALT and AST values were observed, thus indicating that inclusion of cookies containing blueberry and raspberry dried and ground pomace to usual diet might have positive effects on certain cardiovascular risk factors and liver function indicators

Profili masnih kiselina i antioksidativna svojstva sirovih i sušenih oraha

Background: Walnuts consumption produces beneficial effects on human health. Health-promoting benefits are dedicated to its desirable fatty acid profile and high content of antioxidants. Heat treatment of walnuts may alter their fatty acid composition and antioxidant capacity. Aim: In general the aim of this work was to compare fatty acids profiles and antioxidative properties of raw and dried walnuts at 60 °C for 12 hours. Methodology and results: FA profiles were analyzed using gas chromatography. Antioxidative capacities of walnut samples were determined by DPPH and ABTS tests. There were no significant differences in fatty acid profiles comparing dried and raw walnuts. The most abundant fatty acid was linoleic with mean content of 61.38 ± 1.11% in raw and 62.40 ± 0.99% in dried walnuts. Walnuts oil contained 10.64 ± 0.46% and 10.49 ± 0.81% of a-linolenic acid (ALA) in raw and dried walnuts, respectively. Antioxidative capacity of methanolic extracts showed no difference comparing raw and dried walnut by DPPH and ABTS test. Heat treatment at 60 °C for 12h induced no change in fatty acid profiles of walnuts and led to minor decrease in antioxidative capacity measured only by ABTS test. Conclusion: We suggest that drying process in our experiment did not decreased nutritional capacity which is mostly mediated by conservation of fatty acids content in walnuts.Dijetarni unos oraha povezuje se sa brojnim pozitivnim efektima na zdravlje. Svoj efekat na zdravlje orasi ostvaruju zahvaljujući povoljnom masnokiselinskom sastavu i visokom sadržaju antioksidanasa. Zagrevanje oraha moglo bi dovesti do promena u masnokiselinskom sastavu i uticati na antioksidativni kapacitet. Cilj ovog rada bio je da se uporede masnokiselinski profili i antioksidativni status svežih i oraha sušenih na 60 °C u trajanju od 12 sati. Masnokiselinski sastav analiziran je gasnom hromatografijom. Antioksidativni kapaciteti uzoraka oraha odreðivani su DPPH i ABTS testovima. Utvrðeno je da nije bilo promena u masnokiselinskim profilima posle sušenja oraha na 60 °C u trajanju od 12 sati. Najzastupljenija masna kiselina bila je linolna sa sadržajem od 61.38 ± 1.11% u sirovim i 62.40 ± 0.99% kod sušenih oraha. Ulje oraha sadržalo je 10.64 ± 0.46% i 10.49 ± 0.81% a-linolenske kiseline (ALA) u svežim i sušenim orasima, respektivno. DPPH testom nije utvrđeno da postoje razlike u antioksidativnom potencijalu metanolnih ekstrakata oraha sušenih u odnosu na sveže kao ni sa ABTS testom. Naš eksperimet je pokazao da proces sušenja oraha nije smanjio njihov nutritivni kapacitet što je verovatno posredovano očuvanjem sadržaja masnih kiselina

The Effect of Fish Oil-Based Foods on Lipid and Oxidative Status Parameters in Police Dogs

The synthesis, degradation, and reconstruction of the cell membrane as a metabolic pathway of phospholipids is a constant and dynamic process. Fatty acids as bioactive lipid components of plasma and erythrocyte phospholipids as structural lipids have biological roles in the integrity of cell membranes. Fatty acids, depending on the chain length, the degree of saturation, and the synthesis pathways, can alleviate inflammation and oxidative stress caused by excessive exercise. Considering that changing food intake or diet can influence fatty acid phospholipid metabolism, our study aimed to determine the potential benefits of fish-based diets in working (police) dogs undergoing intensive training concerning bioactive lipids such as fatty acids, phospholipids of plasma, and erythrocytes. Fatty acid esters’ composition of plasma and erythrocyte phospholipids as a bioactive lipids, in addition to markers of oxidative stress and metabolic parameters, were analysed by GC chromatography. The food was well tolerated by all dogs, and the compliance to the diet was high throughout the study. After the treatment with fish-based food, blood glucose, total, and LDL cholesterol levels were significantly reduced, indicating positive biochemical profiles of dogs. Correlations of fatty acid phospholipid compositions between plasma and erythrocytes have shown that both plasma and erythrocytes could represent markers of omega-3 eicosapentaenoic and docosahexaenoic acid intake levels in dogs. Morover, fish-based food supplementation caused a significant reduction in lipid peroxidation markers. The enrichment of dogs’ diets with marine fish could improve oxidative status and improve roles and status of bioactive lipids, such as membrane phospholipids and fatty acids, as its components in plasma and erythrocytes in police dogs exposed to intensive exercise

Effects of three types of physical activity on reduction of metabolic parameters involved in cardiovascular risk

The aim of present study was to investigate the effects of three different types of physical activity on reduction of the metabolic parameters mainly responsible for cardiovascular diseases. This prospective-intervention study was performed at the 'ČIGOTA' Thyroid Institute on Mt. Zlatibor (Serbia) between August 2004 and June 2006. Sixty-eight overweight/obese patients aged 40-70 years with hyperlipidemia were divided into three groups according to their weight and overall health. The program of physical workout included: group I - fast walking; group II - gymnastic exercises and specially chosen exercises in the swimming pool; and group III - combined physical training of higher intensity and greater length. All patients were also on a special reduced diet of 1000 kcal per day, the AHA step-2 diet. We monitored the body mass index, body composition, glucose, cholesterol (total, LDL-, and HDL-), and triglycerides before, during, and after the intervention. After 2 and particularly 12 weeks of intervention, a significant improvement of all metabolic parameters was achieved in all three groups of patients. Although most patients completed the study with normal values of all parameters, the most desirable results were achieved in group III (combined exercises with an average energy expenditure of 900 kcal per day). Our research indicates that a specially conceived program of physical activity and diet intervention resulted in significant reduction of cardiovascular risk factors