Browsing by Subject "lipid"

Mercury (Hg) is a heavy metal acknowledged as a worldwide contaminant that accumulates in organisms and biomagnifies in food webs. The organic methylmercury (MeHg) species is harmful to animals, including humans, and mainly derived from the diet. The dietary Hg consumed by fish is mostly removed through the intestine, but some of the MeHg bioaccumulates, especially in the white muscle tissue of fish. Perch (Perca fluviatilis) and roach (Rutilus rutilus) are commonly found fish species in Finland. Perch has additional im-portance as it is the national fish of Finland, a popular food fish, as well as a monitoring species used to evaluate the chemical status of lakes. Seasonal variation of Hg in muscle tissue of fish is supposedly caused by starvation in winter, which condenses Hg in the muscle, and growth dilution in summer, which refers to fast somatic growth during the growing season, which dilutes Hg in the muscle. Similar to winter, spawning has also been found to condense Hg in muscle tissue of fish due to high energy investment into gonad development. Seasonal variation of Hg and variables driving seasonal changes have been shown to differ between fish species. Seasonal variation has been studied mainly during the open-water season. However, less is known about how winter conditions under ice affect Hg levels in fish. In this MSc thesis, I asked (Q1) How total mercury (THg) content in the muscle tissue of perch and roach change annually? (Q2) How THg bioaccumulation in the muscle tissue of perch and roach change annually? (Q3) What are the factors explaining annual variation in THg content in the muscle tissue of perch and roach? The practical application of results was to discuss if annual variation should be considered in monitoring programs and human health questions. The materials used in this study were collected from Lake Pääjärvi monthly from March 2020 to March 2021. Fish were collected using gillnet series. Length, weight, sex, sexual maturity, stomach fullness, Fulton’s condition factor (K), and muscle THg were determined from each fish. The annual length-corrected THg content variation was tested using analysis of variance. The annual THg bioaccumulation variation in the relationship between muscle and fish length was tested using simple linear regression analysis, and the seasonal variation in THg bioaccumulation was tested with LOESS regression analysis. Variables affecting seasonal variation were tested with stepwise multiple linear regression analysis. THg content of perch was the highest in winter and spring and the lowest in fall, while roach showed no significant seasonal variation. THg bioaccumulation of both species was highest in winter, spring, and early summer and lowest in fall. Perch displayed more substantial seasonal variation than roach. Biological and environmental variables that explained the THg content of perch were length, ice thickness, gonadosomatic index (GSI), light, and condition factor. Variables that explained the THg content of roach were length, sex, and total phosphorus (Tot-P). This study confirmed that starvation in winter, growth dilution in summer, and spawning in spring/early summer are vital factors driving seasonal variation. Due to evident seasonal variation, monitoring month should be pre-set in current monitoring programs.

Microvesicles (MVs) are lipid bilayered membranous vesicles containing functional lipids, proteins, RNA and DNA that are produced by most cells. The physiological significance of MVs has become evident, and increased MV counts and the contents of MVs are nowadays also associated with different pathophysiological phenomena. The goal of the field is to use MVs as diagnostic and therapeutic tools. To achieve this, the understanding of the mechanisms of the functions of MVs should be understood better and additionally, reliable methods for the quantification and characterization of MVs should be developed and standardized. The aim of the study was to determine differences in platelet-derived MVs produced by different activation mechanisms. The second aim was to set up and optimize a protocol based on the reaction of sulphur, phosphate and vanillin (SPV) for measuring lipid content of MVs. The third aim was to study the effect of thrombin and proteinase inhibitor PPACK to the vesiculation of platelets. Platelets were isolated from the whole blood of healthy volunteers and vesicles were produced by platelet agonists mediating thrombogenic activation (thrombin and collagen, TC), pathophysiological activation (lipopolysaccharide, LPS) and Ca-ionophore (A23187) as positive control for vesiculation. Quantification and size determination of produced MVs was done using Nanoparticle Tracking Analysis (NTA). MVs were characterized by protein content using bicinchonic acid assay (BCA) and by lipid content using SPV-reaction. MVs had great activation-dependent differences in the lipid and the protein content. Activation with Ca-ionophore produced the most MVs, but the lipid and protein content was only a fraction from (patho)physiologically induced MVs. Only TC increased vesiculation. Vesicle subpopulations had significant difference in lipid content. Thrombin and proteinase inhibitor PPACK mediated inhibition of platelet formation in all of the activations, but the effect was not statistically significant. The mechanism of inhibition was likely to be proteinase inhibitor mediated. The isolation of vesicle populations using differential centrifugation proved to isolate studied populations only partially and the quantification method with NTA was susceptible to concentrated samples. SPV protocol reacted with different intensity to different lipids. In the future, quantification and isolation methods for MVs and the subpopulations of MVs should be improved. Additionally, to understand the physiologically relevant mechanisms of platelet-derived vesicle formation, the inhibitor experiments with PPACK should be continued, because the number of replicates was too low to see significant effects due to a large donor-dependent deviation. Since MVs are heterogenous cellular multitools affecting varying (patho)physiological phenomena, optimization and standardization of methods should be continued in order to study MVs properly.