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Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of its reactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of its reactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Cys34 thiol group of human serum albumin (HSA) represents major plasma antioxidant. Its reactivity is influenced by multiple factors. The influence of fatty acids (FA; saturated, mono, and poly unsaturated acids from fish oil) binding to HSA, on copper(II) binding affinity and Cys34 thiol group accessibility/reactivity, in the presence of carbonylation agent (methylglyoxal, MG) was examined. HSA-copper(II) content, thiol group reactivity, and HSA carbonylation level were monitored spectrophotometrically. Changes in HSA were followed by fluorescence spectroscopy and native PAG electrophoresis. FA/HSA molar ratio was screened by GC. Together, binding of copper(II) ions and FA to HSA increase the reactivity of Cys34 thiol group (depending on the type of FA), with constant contribution of copper(II) ions of one-third. Carbonylation of FA-HSA-Cu(II) complexes caused a decrease in the Cys34 thiol group content, accompanied by a decrease in the content of HSA-bound copper. The carbonylation level of guanidine groups was not affected by FAs and copper(II) binding. Fluorescent emission spectra of FA-HSA-Cu(II)-MG complexes showed conformational changes in HSA molecule. Although binding of fatty acids and copper ions caused a significant increase in the thiol group reactivity, Cys34 thiol from FA-HSA-Cu(II) complexes reacted with MG in smaller extent than expected, probably as a consequence of conformational changes introduced by carbonylation. Increase in the percentage of reacted-free thiol groups with MG (due to FA and copper binding) may not seem to be very significant, but it is very important in complex biological systems, where catalytic metal is present. [GRAPHICS] .

Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of its reactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of itsreactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of its reactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of its reactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of its reactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Cys34 thiol group of human serum albumin (HSA) represents major plasma antioxidant. Its reactivity is influenced by multiple factors. The influence of fatty acids (FA; saturated, mono, and poly unsaturated acids from fish oil) binding to HSA, on copper(II) binding affinity and Cys34 thiol group accessibility/reactivity, in the presence of carbonylation agent (methylglyoxal, MG) was examined. HSA-copper(II) content, thiol group reactivity, and HSA carbonylation level were monitored spectrophotometrically. Changes in HSA were followed by fluorescence spectroscopy and native PAG electrophoresis. FA/HSA molar ratio was screened by GC. Together, binding of copper(II) ions and FA to HSA increase the reactivity of Cys34 thiol group (depending on the type of FA), with constant contribution of copper(II) ions of one-third. Carbonylation of FA-HSA-Cu(II) complexes caused a decrease in the Cys34 thiol group content, accompanied by a decrease in the content of HSA-bound copper. The carbonylation level of guanidine groups was not affected by FAs and copper(II) binding. Fluorescent emission spectra of FA-HSA-Cu(II)-MG complexes showed conformational changes in HSA molecule. Although binding of fatty acids and copper ions caused a significant increase in the thiol group reactivity, Cys34 thiol from FA-HSA-Cu(II) complexes reacted with MG in smaller extent than expected, probably as a consequence of conformational changes introduced by carbonylation. Increase in the percentage of reacted-free thiol groups with MG (due to FA and copper binding) may not seem to be very significant, but it is very important in complex biological systems, where catalytic metal is present. [GRAPHICS] .

Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of its reactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Antipsychotic drugs interfere with the antioxidant defense system provoking complex and often toxicological effects. Here we examined differences in plasma albumin reduced free thiol (SH) group content and its reactivity as a consequence of clozapine (CLZ) and ziprasidone (ZIP) binding. Chronic administration of CLZ reduced, whereas treatment with ZIP increased albumin-SH content in rats. Regardless of the ratio of stearic acid (SA) bound to protein, in vitro binding of ZIP to human serum albumin (HSA) increased both the SH group level and reactivity. In contrast, the effect of CLZ on HSA-SH reactivity was dependent on HSA to SA molar ratio. CLZ binding was accompanied by an increase in HSA-SH reactivity in samples with normal, but a reduction of itsreactivity level with higher SA/HSA ratio, compared to drug-free samples. We demonstrate by steady-state fluorescence quenching studies that an increase in SA binding to HSA is associated with a significant reduction of binding constant for both antipsychotics. In addition, this is the first report of quantitative characterization of ZIP binding to HSA. Our findings suggest that albumin-SH content and reactivity is modulated by ZIP towards an increased antioxidant defense capacity in circulation, as opposed to CLZ, which can contribute to the safer, more effective treatment of schizophrenia.

Gljive iz roda Trichoderma spp. su kosmopolitske filamentozne gljive koje se pojavljuju kao dominantne komponente zemljišnog ekosistema. Ekološka i biološka uloga ovih gljiva je veoma raznovrsna. Kao endofitni simbionti biljaka imaju primenu u kontroli biljnih bolesti čime pospešuju biljni rast i prinos. Takođe, poznata je i njihova sposobnost da indukuju otpornost kod biljaka koje su izložene biotičkom ili abiotičkom stresu. Pored važne primene u poljoprivredi one nalaze primenu i u različitim granama industrije jer proizvode mnogobrojne enzime i metabolite. Obzirom da su mnoge vrste u okviru ovog roda ne mogu razlikovati na morfološkom nivou, neophodna je njihova identifikacja na molekularnom nivou.Cilj ove teze bio je utvrđivanje najvažnijih faktora koji utiču na varijabilnost roda Trichoderma u različitim tipovima zemljišta. Molekularna identifikacija gljiva roda Trichoderma, izolovanih iz različitih tipova zemljišta (rendzina, sirozem na lesu, ritska crnica, fluvisol, ranker, gajnjača, černozem sa različitim podtipovima i smonica) i iz rizosfere različitih biljnih kultura urađena je analizom sekvenci ITS i TEF1-α gena. Uzorci zemljišta ispitani su u funkciji fizičko-hemijskih i mikrobioloških karakteristika, kao i sadržaja ukupnih i lakopristupačnih metala (Cr, Co, Ni, Cu, Zn, Cd, Pb i Mn). U cilju ispitivanja primene ovih gljiva kao antagonista fitopatogenih mikroorganizama, testirani su odabrani izolati prema Fusarium solani species complex (FSSC), Alternaria alternata i Phoma cucurbitacearum. Enzimska karakterizacija izolata odabranih za test antagonizma, urađena je semikvantitativnim testom API-ZYM. Kako bi se ispitao efekat ovih gljiva i na abiotski stres izazvan povećanim sadržajem metala u zemljištu, između ostalih, odabrani su izolati koji su identifikovani iz zemljišta u kojima je određena povećana koncentracija metala, bakra(II) i nikla(II). Na osnovu in vitro testa, gajenjem izolata na povećanim koncentracijama bakra(II) i nikla(II), odabran je izola, Fungi of the genus Trichoderma spp. are cosmopolitan species present in different ecosystems. with important ecological and biological roles.. Beneficial effects of this fungi are responsible for their use in various fields of biotechnology and agriculture. As plant endophytes they are involved in control of plant deseases. They are known to induce plant tolerance to various biotic and abiotic stresses. Known to produce different enzymes, they can be used in preparations of commercial products, such as biopesticides. However, for their adequate use and application their identification on the molecular level is of great importance.The aim of this thesis was to investigate the most important factors that influence Trichoderma variabilty in different soil types. Molecular identification of Trichoderma spp. from different soil types (Rendzic Leptosol, Regosol (Calcaric, Arenic), Calcic Gleysol, Fluvisol, Dystric Leptosol, Eutric Cambisol, Chernozem on loess terrace Chernozem (Arenic) and Vertisol) and plant rhizosphere of different crops was done based on their ITS and TEF1-α sequences. Variability was investigated in function of physical, chemical, microbiological properties of the soil, as well as total and extractable metal (Cr, Co, Ni, Cu, Zn, Cd, Pb and Mn) content in the soil. In order to examine the application of these fungi as antagonists of phytopathogens, selected strains of Trichoderma species were tested against Fusarium solani species complex (FSSC), Alternaria alternata and Phoma cucurbitacearum. The enzyme characterization of selected strains was performed with semiquantitative test API-ZYM. Furthermore, the selected strains were tested for tolerance to various concentrations of copper and nickel. The strain T. brevicompactum SZMC 22661 which showed the best tolerance to metals was applied in the examination of drought effect on tomato plants...

Objective: The objective of the present study was to determine if vitamin D intake and status are associated with pre-eclampsia in a country without a vitamin D fortification policy. Design: A case-control study of pregnancies with (case) and without (control) pre-eclampsia was conducted from January to April when UVB is minimal. Maternal and cord blood obtained at delivery were measured for plasma 25-hydroxycholecalciferol (25-OH-D-3), 3-epimer of 25-OH-D-3 (3-epi-25-OH-D-3) and 24,25-dihydroxycholecalciferol (24,25-(OH)(2)D-3) by LC-MS/MS and maternal 1,25-dihydroxyvitamin D (1,25-(OH) 2D). Differences between groups were tested with ANOVA and Bonferroni post hoc tests (P lt 0.05). Setting: Clinical Center of Serbia. Subjects: Pregnant women with and without pre-eclampsia (n 60) and their infants. Results: Exogenous vitamin D intake (0.95-16.25 mu g/d (38-650 IU/d)) was not significantly different between groups. Women with pre-eclampsia delivered infants at an earlier gestational age and had significantly lower mean total plasma 25-hydroxyvitamin D (25-OH-D; case: 11.2 (SD 5.1); control: 16.1 (SD 5.7) ng/ml; P=0.0006), 25-OH-D-3 (case: 10.0 (SD 4.9); control: 14.2 (SD 5.8) ng/ml; P=0.002), 3-epi-25-OH-D-3 (case: 0.5 (SD 0.2); control: 0.7 (SD 0.2) ng/ml; P=0.0007) and 1,25-(OH)(2)D (case: 56.5 (SD 26.6); control: 81.0 (SD 25.7) pg/ml; P=0.018), while 24,25-(OH)(2)D-3 was not different between groups. Infants did not differ in total plasma 25-OH-D, 25-OH-D-3, 3-epi-25-OH-D-3 and 24,25-(OH)(2)D-3, but the mean proportion of 3-epi-25-OH-D-3 was higher in the infant case group (case: 7.9 (SD 1.1); control: 7.0 (SD 1.4) % of total 25-OH-D-3; P=0.005). Conclusions: A high prevalence of vitamin D deficiency, as defined by plasma 25-OH-D lt 12 ng/ml, was observed in 47 % of all mothers and 77 % of all infants. These data underscore the need for prenatal vitamin D supplementation and a food fortification policy in Serbia.

Objective: The objective of the present study was to determine if vitamin D intake and status are associated with pre-eclampsia in a country without a vitamin D fortification policy. Design: A case-control study of pregnancies with (case) and without (control) pre-eclampsia was conducted from January to April when UVB is minimal. Maternal and cord blood obtained at delivery were measured for plasma 25-hydroxycholecalciferol (25-OH-D-3), 3-epimer of 25-OH-D-3 (3-epi-25-OH-D-3) and 24,25-dihydroxycholecalciferol (24,25-(OH)(2)D-3) by LC-MS/MS and maternal 1,25-dihydroxyvitamin D (1,25-(OH) 2D). Differences between groups were tested with ANOVA and Bonferroni post hoc tests (P lt 0.05). Setting: Clinical Center of Serbia. Subjects: Pregnant women with and without pre-eclampsia (n 60) and their infants. Results: Exogenous vitamin D intake (0.95-16.25 mu g/d (38-650 IU/d)) was not significantly different between groups. Women with pre-eclampsia delivered infants at an earlier gestational age and had significantly lower mean total plasma 25-hydroxyvitamin D (25-OH-D; case: 11.2 (SD 5.1); control: 16.1 (SD 5.7) ng/ml; P=0.0006), 25-OH-D-3 (case: 10.0 (SD 4.9); control: 14.2 (SD 5.8) ng/ml; P=0.002), 3-epi-25-OH-D-3 (case: 0.5 (SD 0.2); control: 0.7 (SD 0.2) ng/ml; P=0.0007) and 1,25-(OH)(2)D (case: 56.5 (SD 26.6); control: 81.0 (SD 25.7) pg/ml; P=0.018), while 24,25-(OH)(2)D-3 was not different between groups. Infants did not differ in total plasma 25-OH-D, 25-OH-D-3, 3-epi-25-OH-D-3 and 24,25-(OH)(2)D-3, but the mean proportion of 3-epi-25-OH-D-3 was higher in the infant case group (case: 7.9 (SD 1.1); control: 7.0 (SD 1.4) % of total 25-OH-D-3; P=0.005). Conclusions: A high prevalence of vitamin D deficiency, as defined by plasma 25-OH-D lt 12 ng/ml, was observed in 47 % of all mothers and 77 % of all infants. These data underscore the need for prenatal vitamin D supplementation and a food fortification policy in Serbia.

Humani serum albumin (HSA) je najzastupljeniji protein plazme sa udelomod oko 50 do 60 % svih proteina plazme. Vezuje i transportuje mnoge endogene iegzogene molekule, i doprinosi antioksidativnom kapacitetu seruma jer na površinimolekula poseduje jednu slobodnu tiolnu grupu ostatka Cys34. Slobodne masne kiseline(MK), koje transportuje HSA, i karbonilni stres mogli bi, kroz promenu reaktivnostitiolne grupe, uticati na njen potencijal kao hvatača reaktivnih karbonilnih vrsta. Stogasu ciljevi ove teze bili: in vitro ispitivanje uticaja 1) vezivanja MK, različite dužinelanca i zasićenosti na (i) reaktivnost Cys34-SH grupe, (ii) njen potencijal kao hvatačareaktivnih α-oksoaldehida, tj. na stepen karbonilacije i (iii) reaktivnost tiolne grupeHSA karbonilovanog metilglioksalom (model-sistem za molekule HSA modifikovane ukarbonilnom stresu); 2) vezivanja MK ribljeg ulja na reaktivnost Cys34-SH grupe(sagledavanje mogućnosti modulacije svojstava HSA pomoću suplemenata); 3)određivanje sadržaja Cys34-SH grupe, korelacija sa sadržajem HbA1c i glukoze ukarbonilnim stresom; 4) određivanje reaktivnosti Cys34-SH grupe HSA izolovanog izseruma dijabetičara i zdravih osoba; 5) razvijanje qTLC metode za određivanje sadržajaMK, vezanih za HSA, koji je izolovan iz realnih uzoraka.Vezivanje MK (različite dužine lanca i zasićenosti: miristinske (MYR),palmitinske (PLM), stearinske (STE), oleinske (OLA), eikozapentenske (EPA) idokozaheksaenske kiseline (DHA)) za HSA in vitro, dovodi do povećanja vrednostikonstanti brzine reakcije (k) Cys34 tiolne grupe (i DTNB) (k vrednosti kompleksa HSAMK:od 14,58±0,19 x 10-3 do 26,02±1,06 x 10-3 s-1 u odnosu na k HSA: 7,52±0,04 x 10-3 s-1), odnosno njene reaktivnosti za 2 do 3,5 puta. STE i OLA ispoljavaju slične efektena reaktivnost HSA-SH, a najjači efekat ispoljava polinezasićena EPA. Od zasićenihMK, najveća vrednost k dobijena je pri vezivanju MYR za HSA, što može biti posledicaizuzetne komplementarnosti njenog molekula sa vezivnim mestima na molekulu, Human serum albumin (HSA) is the most abundant protein of humanplasma, accounting for 50-60% of total plasma proteins. HSA binds and transport manyendogenous and exogenous substances, and contributes to the antioxidative pool inserum because of Cys34 free thiol group on the surface of its molecule. Free fatty acids(FAs), transported by HSA, and carbonyl stress could influence, through changedreactivity of the thiol group, on its potential as the scavenger of the reactive carbonylspecies. Therefore, the aims of this thesis were: in vitro examination of the influence 1)of different (in terms of chain length and saturation) FAs binding on the: (i) reactivity ofthe Cys34-SH group, (ii) its potential as the scavenger of the reactive α-oxoladehydes,and (iii) on the reactivity of the thiol group of HSA carbonylated with methylglyoxal(the model-system for HSA molecules modified during carbonyl stress); 2) of FAsfrom fish oil on the Cys34-SH group reactivity (overviewing the possibility of themodulation of the HSA scavenger properties with supplements); 3) determination of theCys34-SH group content, correlation with the HbA1c and glucose level of persons withelevated carbonyl stress; 4) determination of the Cys34-SH group reactivity of the HSAisolated from the sera of diabetic patients and healthy persons; 5) development of qTLCmethod for determination of the FAs content, bound to the HSA, which was isolatedfrom sera samples.Binding of FAs of different chain lengths and saturation (myristic (MYR),palmitic (PLM), stearic (STE), oleic (OLA), eicosapentaenoic (EPE) anddocosahexaenoic acid (DHA)) to the HSA in vitro resulting in increasing of the kineticsconstant (k) of the Cys34 thiol group reaction (with DTNB)(k values for HSA-FAscomplexes: from 14.58±0.19 ×10-3 to 26.02±1.06 × 10-3 s-1 comparing to k of the HSAsolely 7.52 ± 0.04 × 10-3 s-1), i.e. its reactivity for 2 to 3.5 times. STE and OLA showsimilar effects to the HSA-SH reactivity, and the strongest effect shows polyuns

Objective: The objective of the present study was to determine if vitamin D intake and status are associated with pre-eclampsia in a country without a vitamin D fortification policy. Design: A case-control study of pregnancies with (case) and without (control) pre-eclampsia was conducted from January to April when UVB is minimal. Maternal and cord blood obtained at delivery were measured for plasma 25-hydroxycholecalciferol (25-OH-D-3), 3-epimer of 25-OH-D-3 (3-epi-25-OH-D-3) and 24,25-dihydroxycholecalciferol (24,25-(OH)(2)D-3) by LC-MS/MS and maternal 1,25-dihydroxyvitamin D (1,25-(OH) 2D). Differences between groups were tested with ANOVA and Bonferroni post hoc tests (P lt 0.05). Setting: Clinical Center of Serbia. Subjects: Pregnant women with and without pre-eclampsia (n 60) and their infants. Results: Exogenous vitamin D intake (0.95-16.25 mu g/d (38-650 IU/d)) was not significantly different between groups. Women with pre-eclampsia delivered infants at an earlier gestational age and had significantly lower mean total plasma 25-hydroxyvitamin D (25-OH-D; case: 11.2 (SD 5.1); control: 16.1 (SD 5.7) ng/ml; P=0.0006), 25-OH-D-3 (case: 10.0 (SD 4.9); control: 14.2 (SD 5.8) ng/ml; P=0.002), 3-epi-25-OH-D-3 (case: 0.5 (SD 0.2); control: 0.7 (SD 0.2) ng/ml; P=0.0007) and 1,25-(OH)(2)D (case: 56.5 (SD 26.6); control: 81.0 (SD 25.7) pg/ml; P=0.018), while 24,25-(OH)(2)D-3 was not different between groups. Infants did not differ in total plasma 25-OH-D, 25-OH-D-3, 3-epi-25-OH-D-3 and 24,25-(OH)(2)D-3, but the mean proportion of 3-epi-25-OH-D-3 was higher in the infant case group (case: 7.9 (SD 1.1); control: 7.0 (SD 1.4) % of total 25-OH-D-3; P=0.005). Conclusions: A high prevalence of vitamin D deficiency, as defined by plasma 25-OH-D lt 12 ng/ml, was observed in 47 % of all mothers and 77 % of all infants. These data underscore the need for prenatal vitamin D supplementation and a food fortification policy in Serbia.

Objective: The objective of the present study was to determine if vitamin D intake and status are associated with pre-eclampsia in a country without a vitamin D fortification policy. Design: A case-control study of pregnancies with (case) and without (control) pre-eclampsia was conducted from January to April when UVB is minimal. Maternal and cord blood obtained at delivery were measured for plasma 25-hydroxycholecalciferol (25-OH-D-3), 3-epimer of 25-OH-D-3 (3-epi-25-OH-D-3) and 24,25-dihydroxycholecalciferol (24,25-(OH)(2)D-3) by LC-MS/MS and maternal 1,25-dihydroxyvitamin D (1,25-(OH) 2D). Differences between groups were tested with ANOVA and Bonferroni post hoc tests (P lt 0.05). Setting: Clinical Center of Serbia. Subjects: Pregnant women with and without pre-eclampsia (n 60) and their infants. Results: Exogenous vitamin D intake (0.95-16.25 mu g/d (38-650 IU/d)) was not significantly different between groups. Women with pre-eclampsia delivered infants at an earlier gestational age and had significantly lower mean total plasma 25-hydroxyvitamin D (25-OH-D; case: 11.2 (SD 5.1); control: 16.1 (SD 5.7) ng/ml; P=0.0006), 25-OH-D-3 (case: 10.0 (SD 4.9); control: 14.2 (SD 5.8) ng/ml; P=0.002), 3-epi-25-OH-D-3 (case: 0.5 (SD 0.2); control: 0.7 (SD 0.2) ng/ml; P=0.0007) and 1,25-(OH)(2)D (case: 56.5 (SD 26.6); control: 81.0 (SD 25.7) pg/ml; P=0.018), while 24,25-(OH)(2)D-3 was not different between groups. Infants did not differ in total plasma 25-OH-D, 25-OH-D-3, 3-epi-25-OH-D-3 and 24,25-(OH)(2)D-3, but the mean proportion of 3-epi-25-OH-D-3 was higher in the infant case group (case: 7.9 (SD 1.1); control: 7.0 (SD 1.4) % of total 25-OH-D-3; P=0.005). Conclusions: A high prevalence of vitamin D deficiency, as defined by plasma 25-OH-D lt 12 ng/ml, was observed in 47 % of all mothers and 77 % of all infants. These data underscore the need for prenatal vitamin D supplementation and a food fortification policy in Serbia.

Objective: The objective of the present study was to determine if vitamin D intake and status are associated with pre-eclampsia in a country without a vitamin D fortification policy. Design: A case-control study of pregnancies with (case) and without (control) pre-eclampsia was conducted from January to April when UVB is minimal. Maternal and cord blood obtained at delivery were measured for plasma 25-hydroxycholecalciferol (25-OH-D-3), 3-epimer of 25-OH-D-3 (3-epi-25-OH-D-3) and 24,25-dihydroxycholecalciferol (24,25-(OH)(2)D-3) by LC-MS/MS and maternal 1,25-dihydroxyvitamin D (1,25-(OH) 2D). Differences between groups were tested with ANOVA and Bonferroni post hoc tests (P lt 0.05). Setting: Clinical Center of Serbia. Subjects: Pregnant women with and without pre-eclampsia (n 60) and their infants. Results: Exogenous vitamin D intake (0.95-16.25 mu g/d (38-650 IU/d)) was not significantly different between groups. Women with pre-eclampsia delivered infants at an earlier gestational age and had significantly lower mean total plasma 25-hydroxyvitamin D (25-OH-D; case: 11.2 (SD 5.1); control: 16.1 (SD 5.7) ng/ml; P=0.0006), 25-OH-D-3 (case: 10.0 (SD 4.9); control: 14.2 (SD 5.8) ng/ml; P=0.002), 3-epi-25-OH-D-3 (case: 0.5 (SD 0.2); control: 0.7 (SD 0.2) ng/ml; P=0.0007) and 1,25-(OH)(2)D (case: 56.5 (SD 26.6); control: 81.0 (SD 25.7) pg/ml; P=0.018), while 24,25-(OH)(2)D-3 was not different between groups. Infants did not differ in total plasma 25-OH-D, 25-OH-D-3, 3-epi-25-OH-D-3 and 24,25-(OH)(2)D-3, but the mean proportion of 3-epi-25-OH-D-3 was higher in the infant case group (case: 7.9 (SD 1.1); control: 7.0 (SD 1.4) % of total 25-OH-D-3; P=0.005). Conclusions: A high prevalence of vitamin D deficiency, as defined by plasma 25-OH-D lt 12 ng/ml, was observed in 47 % of all mothers and 77 % of all infants. These data underscore the need for prenatal vitamin D supplementation and a food fortification policy in Serbia.

Objective: The objective of the present study was to examine the external validity of an FFQ designed to estimate dietary vitamin D intake compared with a plasma biomarker and three repeated 24 h dietary recalls in women of reproductive age in Serbia, where there is no exposure to food fortified with vitamin D. The method of triads was applied. Design: In a cross-sectional study, 422 women completed the Women and Reproductive Health FFQ (WRH-FFQ) during the winter months. From a representative subgroup (n 44), three 24 h dietary recalls and anthropometric parameters were collected as well as a fasting blood sample for vitamin D biomarker analyses. Correlation coefficients were calculated between each of the dietary methods. Validity coefficients, as a correlation between the measured and estimated 'true' exposure, were calculated using the method of triads. Bland-Altman plots were also constructed. Setting: Three major universities in Serbia. Subjects: Healthy young women (n 422) aged 18-35 years. Results: The WRH-FFQ estimate of vitamin D intake for all participants was 4.0 (SD 3.3) mu g/d and 3.1 (SD 2.3) mu g/d for the subgroup. Bland-Altman plots for these intakes showed high agreement. Validity coefficients for the FFQ, 24 h recall and biomarker were. rho(QI) = 0.847 (95 % CI 0.564, 0.928), rho(RI) = 0.810 (95 % CI 0.537, 0.997) and rho(BI) = 0.499 (95 % CI 0.190, 0.840), while the correlation coefficients were 0.686, 0.422 and 0.404. Conclusions: The FFQ applied in the present study is a valid tool for assessing dietary vitamin D intake in women living in Serbia, a region without mandatory vitamin D food fortification.

Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states. (C) 2016 Elsevier B.V. All rights reserved.

This paper describes the results achieved by high school seniors on an item which involves translation of the equation P = kT into a corresponding pictorial external representation. The majority of students (the classes of 2011, 2012 and 2013) did not give the correct answer to the multiple choice part of the translation item. They chose pictorial representations of the other gas laws (P = k/V, or V = kT) instead. Failure to choose the correct answer was surprising considering that the symbol for volume was absent which should have been the key clue. Through the analysis of students' explanations (the classes of 2011 and 2012) and interviews (the class of 2013) we considered the reasoning applied by students who chose the correct answer or distractors for the multiple choice part of the item. Among the students who answered correctly there were explanations which contained misconceptions. Several factors that lead to the unsuccessful translation between external representations have been discovered. Students interpreted the change in one quantity based on the notation for the change in another one because of deep rooted cognitive schemas about changing two quantities (volume and pressure, pressure and temperature, temperature and volume), without consideration that for such changes to be valid for gases all three quantities need to be considered for a certain amount of substance. Those cognitive schemas interfered with mathematical reasoning, i.e. students possessed limited understanding of the equations.

Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states. (C) 2016 Elsevier B.V. All rights reserved.

This paper describes the results achieved by high school seniors on an item which involves translation of the equation P = kT into a corresponding pictorial external representation. The majority of students (the classes of 2011, 2012 and 2013) did not give the correct answer to the multiple choice part of the translation item. They chose pictorial representations of the other gas laws (P = k/V, or V = kT) instead. Failure to choose the correct answer was surprising considering that the symbol for volume was absent which should have been the key clue. Through the analysis of students' explanations (the classes of 2011 and 2012) and interviews (the class of 2013) we considered the reasoning applied by students who chose the correct answer or distractors for the multiple choice part of the item. Among the students who answered correctly there were explanations which contained misconceptions. Several factors that lead to the unsuccessful translation between external representations have been discovered. Students interpreted the change in one quantity based on the notation for the change in another one because of deep rooted cognitive schemas about changing two quantities (volume and pressure, pressure and temperature, temperature and volume), without consideration that for such changes to be valid for gases all three quantities need to be considered for a certain amount of substance. Those cognitive schemas interfered with mathematical reasoning, i.e. students possessed limited understanding of the equations.

Objective: The objective of the present study was to examine the external validity of an FFQ designed to estimate dietary vitamin D intake compared with a plasma biomarker and three repeated 24 h dietary recalls in women of reproductive age in Serbia, where there is no exposure to food fortified with vitamin D. The method of triads was applied. Design: In a cross-sectional study, 422 women completed the Women and Reproductive Health FFQ (WRH-FFQ) during the winter months. From a representative subgroup (n 44), three 24 h dietary recalls and anthropometric parameters were collected as well as a fasting blood sample for vitamin D biomarker analyses. Correlation coefficients were calculated between each of the dietary methods. Validity coefficients, as a correlation between the measured and estimated 'true' exposure, were calculated using the method of triads. Bland-Altman plots were also constructed. Setting: Three major universities in Serbia. Subjects: Healthy young women (n 422) aged 18-35 years. Results: The WRH-FFQ estimate of vitamin D intake for all participants was 4.0 (SD 3.3) mu g/d and 3.1 (SD 2.3) mu g/d for the subgroup. Bland-Altman plots for these intakes showed high agreement. Validity coefficients for the FFQ, 24 h recall and biomarker were. rho(QI) = 0.847 (95 % CI 0.564, 0.928), rho(RI) = 0.810 (95 % CI 0.537, 0.997) and rho(BI) = 0.499 (95 % CI 0.190, 0.840), while the correlation coefficients were 0.686, 0.422 and 0.404. Conclusions: The FFQ applied in the present study is a valid tool for assessing dietary vitamin D intake in women living in Serbia, a region without mandatory vitamin D food fortification.

Vitamin D je od izuzetne važnosti u trudnoći pre svega za normalan rast kostiju i zacelokupan razvoj bebe. Pokazano je međutim da 40-80 % trudnica ima nizak nivo (< 20ng/ml) 25-hidroksivitamina D (25(OH)D) u serumu kao i da nizak nivo vitamina D utrudnoći povećava rizik za pojavu preeklampsije (PE) i prevremenih porođaja. U Srbiji,gde hrana nije obogaćena vitaminom D, stopa preeklampsije je veća u zimskom periodu.Odgovor na suplemenatciju vitaminom D dodatno se usložnjava prisustvom C-3α-epimera 25(OH)D3 (3-epi-25(OH)D3), čiji udeo u ukupnom nivou 25(OH)D u krvi možebiti i do 60 % kod beba, a do 17 % kod odraslih. Prisustvo 3-epi-25(OH)D3 u krvi pupčanevrpce ukazuje da postoji prenos sa majke na novorođenče. Međutim, i dalje je nepoznatokoja je uloga povišenog nivoa 3-epi-25(OH)D3 kod novorođenčadi, kako se nivo menjatokom prvih nedelja života novorođenčadi kao i kakav efekat 3-epi-25(OH)D3 ima na rastkostiju beba rođenih iz fizioloških trudnoća u odnosu na trudnoće sa komplikacijama kaošto je preeklampsija. Stoga su ciljevi ove teze: 1) validacija upitnika o učestalostikonzumiranja namirnica (WRH-FFQ) koji bi omogućio procenu unosa vitamina D kodmladih žena u reproduktivnom dobu u Srbiji; 2) sagledavanje povezanosti niskog unosa istatusa vitamina D sa pojavom preeklampsije, kao i određivanje udela 3-epi-25(OH)D3 uukupnom sadržaju 25(OH)D3 u odnosu na zdravstvene ishode majki i novorođenčadi; 3)određivanje nivoa 3-epi-25(OH)D3 tokom prvih nedelja života novorođenčadi isagledavanje njegovog efekta na rast kostiju beba rođenih iz fizioloških trudnoća uodnosu na trudnoće sa komplikacijama kao što je preeklampsija; i 4) ispitivanje biološkogodgovora kostiju na izloženost endogenom i egzogenom izvoru 3-epi-25(OH)D3 naanimalnom modelu.Studija validacije upitnika WRH-FFQ za procenu unosa vitamina D kod mladih žena ureproduktivnom dobu u Srbiji izvedena je od novembra do aprila, kada je smanjeno UVBzračenje. Upitnik WRH-FFQ adekvatno su popunile 422 žene od kojih su nasumič, Vitamin D exposure during pregnancy is important for skeletal growth and developmentin the offspring. However, 40-80% of expecting mothers have low (< 20 ng/ml) serum25-hydroxyvitamin D (25(OH)D) level . A recent meta-analysis demonstrated that lowvitamin D status in pregnancy increases the risk of preeclampsia (PE) and preterm birth.In Serbia where there is no mandatory vitamin D food fortification policy, rates of PEare higher in winter. Response to vitamin D supplementation is complicated by thepresence of C-3α epimer of 25(OH)D3 (3-epi-25(OH)D3) representing up to 60% oftotal 25(OH)D in infants and 17% in adults, raising questions regarding its biologicalderivation and function. The presence of cord 3-epi-25(OH)D3 suggests maternal-fetaltransfer. However, it is still unknown whether 3-epi-25(OH)D3 increases in the first fewweeks after birth and how it effects bone mineral accretion in infants of healthy andcomplicated pregnancies. Therefore, sequential objectives of this thesis are to: 1)validate a food frequency questionnaire (WRH-FFQ) for assessing vitamin D intake inyoung women of reproductive age in Serbia; 2) determine if low vitamin D intake andstatus is associated with PE in Serbia; and describe the proportion of total 25(OH)D inthe 3-epi-25(OH)D3 form in relation to maternal and neonatal outcomes; 3) establish if3-epi-25(OH)D3 in preterm and term infants of mothers with and without PE ispredictive of bone mass at term age using a nested case vs. control design; and 4) testthe biological response of bone to endogenous and exogenous 3-epi-25(OH)D3 in ananimal model.The Women and Reproductive Health FFQ (WRH-FFQ) was completed by 422 women.A cross-sectional study, was conducted from November to April when UVB in Serbia isminimal. On a representative sub-sample (n=44) three 24 hour dietary recalls andanthropometric parameters were collected. Fasting blood samples for vitamin Dbiomarkers analyses were available for 23 women...

Poznato je da su semenke lana, susama i golice bogate esencijalnim masnim kiselinama ida predstavljaju značajni dijetarni izvor vitamina E i polifenola. Kod osoba sa hroničnombubrežnom insuficijencijom, na programu hemodijalize, aktuelna su ispitivanja dijetarnih mera ucilju smanjenja incidence kardiovaskularnih oboljenja. Iz navedenog proizišao je cilj ovedisertacije da se ispitaju efekti suplementacije ishrane smešom semenki lana, susama i golice kodbolesnika na hemodijalizi.Antioksidativna uloga humanog serum-albumina (HSA) u značajanoj meri određujeukupnu antioksidativnu aktivnost u plazmi, a analiza HSA pokazala je da je kod bolesnika naprogramu hemodijalize smanjen sadržaj redukovane Cys34 tiolne grupe (od koje potičeantioksidativni potencijal HSA) i povećan stepen karbonilacije HSA. Stoga je drugi cilj ovedisertacije bio da se in vitro ispita uticaj vezivanja komponenti ekstrakta smeše semenki lana,susama i golice, ferulinske kiseline (FE) i metabolita lignana (enterolaktona (EL) i enterodiola(ED), koji se povećavaju u cirkulaciji nakon konzumiranja namirnica bogatih biljnim lignanima),kao i masnih kiselina, na promenu konformacije HSA i reaktivnosti (antioksidativnogpotencijala) Cys34 tiolne grupe, a da bi se sagledali mogući mehanizmi kojima komponente lana,susama i golice ostvaruju svoje in vivo efekte, pre svega antioksidativno dejstvo.Analizom masnokiselinskih profila ukupnih lipidnih ekstrakata smeše semenki gasnotečnomhromatografijom potvrđeno da je u semenkama lana najzastupljenija α-linolenskakiselina (ALA), a u semenkama susama i golice linolna kiselina (LA). Hladnoceđena uljaispoljila su različitu antioksidativnu aktivnost (rafinisano suncokretovo ulje>maslinovo ulje>uljelana>ulje golice> ulje susama) pri DPPH testu, u opsegu od 970 do 2000 μmola TE/1000 g ulja.Sadržaj hidrofilnih antioksidanasa u hladnoceđenim uljima je veoma nizak, sem za hladoceđenomaslinovo ulje. Pored lipofilnih antioksidanasa, pokazano je da metanolni ekstraktiobezmašćenih se, It is known that seeds of flax, sesame, and pumpkin are reach in essential fatty acids, andrepresent significant dietary source of vitamin E and polyphenols. In patients with chronic renalinsufficiency, on hemodialysis treatment, research topics about dietary supplementations withaims to reduce incidence of cardiovascular diseases, are actual. Therefore, the goal of thisdissertation was to investigate the effects of diet supplementation with the mixture of flax,sesame, and pumpkin seeds in patients on hemodialysis.The total antioxidative activity in plasma is significantly determined by antioxidative roleof human serum albumin (HSA), and analysis of the HSA in patients on hemodialysis showedthat the content of the free Cys34 thiol group (from which the HSA antioxidative potentialoriginates) is reduced and on the other hand, degree of HSA carbonylation is increased. Thus, thesecond goal of this dissertation was in vitro investigation of the binding of flax, sesame, andpumpkins seeds extracts components, ferulic acid (FE) and lignan metabolites (enterolactone(EL), enterodiol (ED), which are increased in circulation after consumption of food reach withplants lignan), as well as fatty acids, on the conformational change of the HSA and on thereactivity (antioxidative potential) of Cys34 thiol group, with the final goal to perceive thepossible mechanisms with which the flax, sesame, and pumpkin seeds explore in vivoantioxidative effects.Analysis of fatty acids profiles of total lipid extracts of seeds mixture using gas-liquidchromatography, confirmed that flaxseeds are the most abundant with α-linolenic acid (ALA),and sesame and pumpkin with linoleic acid (LA). Cold pressed oils showed differentantioxidative activity (refined sunflower oil>olive oil>flaxseed oil>pumpkin oil>sesame oil) inthe DPPH test (ranged from 970 to 2000 μmol TE/1000g oil). Content of hydrophilicantioxidants in cold pressed oils was extremely low, except for cold pressed olive oil. Besideslipophilic

Funkcije membranskih proteina su brojne: međućelijska komunikacija, adhezija, signalna transdukcija. Većina membranskih proteina je glikozilovana i N-glikani imaju važnu ulogu u formiranju trodimenzionalne strukture membranskih proteina, kao i u ispoljavanju njihove funkcije. Receptori za insulin (IR) i faktor rasta sličan insulinu tip 1 (IGF1R) su transmembranske tirozin-kinaze sa visokim stepenom homologije (u nekim domenima čak i 80%). Familija IGF receptora, pored IR i IGF1R, uključuje i receptor za faktore rasta slične insulinu tip 2 (IGF2R). Sva tri receptora (IR, IGF1R i IGF2R) su glikozilovani i obilno prisutni u placenti, gde imaju važne uloge u njenom razvoju i funkcionisanju.Placenta raste i razvija se da bi ispunila različite potrebe fetusa, pa se struktura i funkcija placente menjaju tokom gestacije. Znajući da su proteini odgovorni za biološke funkcije placente pretpostavljeno je da tokom gestacije može doći i do promene u sadržaju različitih tipova N-glikana prisutnih na membranskim proteinima. U ovoj tezi analizirani su tipovi N-glikana koji se mogu naći u sastavu membranskih glikoproteina, odnosno membranski N-glikom proteina humane placente. Ispitana je podložnost membranskog N-glikoma individualnim varijacijama i uticaju starosti majke/trudnice, kao i uticaj gestacije na membranski N-glikom. Znajući da je izmenjena glikozilacija često povezana sa izmenjenom funkcijom, kao i da izmenjena funkcija jednog ili više proteina može biti uzrok bolesti, ispitane su potencijalne promene membranskog N-glikoma u patološkim trudnoćama (kod preeklampsije) i trudnoćama komplikovanim patologijom majki (dijabetes). Uporedo je ispitan i uticaj starosti, gestacije i patologije na tip i zastupljenost različitih N-glikana koji ulaze u sastav receptora IGF sistema. Cilj je bio da se ispita da li promene na ukupnim membranskim proteinima (na N-glikomu) prate promene N-glikana prisutnih na pojedinačnim membranskim glikoproteinima važnim za rast i funkcionisanje placente.

The interaction of polyphenolic molecules with human serum albumin (HSA) could lead to changes in the reactivity of the HSA Cys34 thiol group (HSA-SH). The influences of enterolactone (EL) and enterodiol (ED) binding on HSA-SH reactivity in fatty acid (FA)-free HSA, and in HSA with bound stearic acid (S) in S/HSA molar ratios of 1 : 1 and 4 : 1, were investigated by the determination of the pseudo first order rate constants (k') for the thiol reaction with 5,5'-dithiobis-(2-nitrobenzoic acid). The binding affinities and binding sites of EL and ED were also determined, using fluorescence measurements of the intrinsic fluorescence of Trp214 and diazepam (binding site marker). EL and ED binding to HSA increased the reactivity of HSA-SH in all assayed HSA-enterolignan complexes by 9.1-33.1%. The strongest effects were obtained for FA-free HSA-enterolignan complexes. S modulated/reduced the effect of EL on HSA-SH reactivity, while its influence on the effect of ED was negligible. The binding of enterolignans to HSA was investigated: the binding constants were the highest for FA-free HSA (EL: 11.64 x 10(4) M-1 and ED: 5.59 x 10(4) M-1 at 37 degrees C) and the lowest for S/HSA 4 : 1-enterolignan complexes (EL: 2.43 x 10(4) M-1 and ED: 1.92 x 10(4) M-1). When the S/HSA ratio was increased, the binding affinities and number of binding sites for EL and ED were decreased. At the same time, a high correlation between binding constants and increased Cys34 reactivity was found (r = 0.974). Competitive experiments using diazepam indicated that the binding of ED and of EL was located in the hydrophobic pocket of site II in HSA. Overall, it is evident that stearic acid could modulate the enterolignan effects on HSA-SH reactivity as well as their binding to HSA. This finding could be important for pharmacokinetics and the expression of enterolignan antioxidant effects in vivo after an intake of lignan rich food.

Objective: The objective of the present study was to examine the external validity of an FFQ designed to estimate dietary vitamin D intake compared with a plasma biomarker and three repeated 24 h dietary recalls in women of reproductive age in Serbia, where there is no exposure to food fortified with vitamin D. The method of triads was applied. Design: In a cross-sectional study, 422 women completed the Women and Reproductive Health FFQ (WRH-FFQ) during the winter months. From a representative subgroup (n 44), three 24 h dietary recalls and anthropometric parameters were collected as well as a fasting blood sample for vitamin D biomarker analyses. Correlation coefficients were calculated between each of the dietary methods. Validity coefficients, as a correlation between the measured and estimated 'true' exposure, were calculated using the method of triads. Bland-Altman plots were also constructed. Setting: Three major universities in Serbia. Subjects: Healthy young women (n 422) aged 18-35 years. Results: The WRH-FFQ estimate of vitamin D intake for all participants was 4.0 (SD 3.3) mu g/d and 3.1 (SD 2.3) mu g/d for the subgroup. Bland-Altman plots for these intakes showed high agreement. Validity coefficients for the FFQ, 24 h recall and biomarker were. rho(QI) = 0.847 (95 % CI 0.564, 0.928), rho(RI) = 0.810 (95 % CI 0.537, 0.997) and rho(BI) = 0.499 (95 % CI 0.190, 0.840), while the correlation coefficients were 0.686, 0.422 and 0.404. Conclusions: The FFQ applied in the present study is a valid tool for assessing dietary vitamin D intake in women living in Serbia, a region without mandatory vitamin D food fortification.

Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states.

The interaction of polyphenolic molecules with human serum albumin (HSA) could lead to changes in the reactivity of the HSA Cys34 thiol group (HSA-SH). The influences of enterolactone (EL) and enterodiol (ED) binding on HSA-SH reactivity in fatty acid (FA)-free HSA, and in HSA with bound stearic acid (S) in S/HSA molar ratios of 1 : 1 and 4 : 1, were investigated by the determination of the pseudo first order rate constants (k') for the thiol reaction with 5,5'-dithiobis-(2-nitrobenzoic acid). The binding affinities and binding sites of EL and ED were also determined, using fluorescence measurements of the intrinsic fluorescence of Trp214 and diazepam (binding site marker). EL and ED binding to HSA increased the reactivity of HSA-SH in all assayed HSA-enterolignan complexes by 9.1-33.1%. The strongest effects were obtained for FA-free HSA-enterolignan complexes. S modulated/reduced the effect of EL on HSA-SH reactivity, while its influence on the effect of ED was negligible. The binding of enterolignans to HSA was investigated: the binding constants were the highest for FA-free HSA (EL: 11.64 x 10(4) M-1 and ED: 5.59 x 10(4) M-1 at 37 degrees C) and the lowest for S/HSA 4 : 1-enterolignan complexes (EL: 2.43 x 10(4) M-1 and ED: 1.92 x 10(4) M-1). When the S/HSA ratio was increased, the binding affinities and number of binding sites for EL and ED were decreased. At the same time, a high correlation between binding constants and increased Cys34 reactivity was found (r = 0.974). Competitive experiments using diazepam indicated that the binding of ED and of EL was located in the hydrophobic pocket of site II in HSA. Overall, it is evident that stearic acid could modulate the enterolignan effects on HSA-SH reactivity as well as their binding to HSA. This finding could be important for pharmacokinetics and the expression of enterolignan antioxidant effects in vivo after an intake of lignan rich food.

Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states. (C) 2016 Elsevier B.V. All rights reserved.

This paper describes the results achieved by high school seniors on an item which involves translation of the equation P = kT into a corresponding pictorial external representation. The majority of students (the classes of 2011, 2012 and 2013) did not give the correct answer to the multiple choice part of the translation item. They chose pictorial representations of the other gas laws (P = k/V, or V = kT) instead. Failure to choose the correct answer was surprising considering that the symbol for volume was absent which should have been the key clue. Through the analysis of students' explanations (the classes of 2011 and 2012) and interviews (the class of 2013) we considered the reasoning applied by students who chose the correct answer or distractors for the multiple choice part of the item. Among the students who answered correctly there were explanations which contained misconceptions. Several factors that lead to the unsuccessful translation between external representations have been discovered. Students interpreted the change in one quantity based on the notation for the change in another one because of deep rooted cognitive schemas about changing two quantities (volume and pressure, pressure and temperature, temperature and volume), without consideration that for such changes to be valid for gases all three quantities need to be considered for a certain amount of substance. Those cognitive schemas interfered with mathematical reasoning, i.e. students possessed limited understanding of the equations.

Objective: The objective of the present study was to examine the external validity of an FFQ designed to estimate dietary vitamin D intake compared with a plasma biomarker and three repeated 24 h dietary recalls in women of reproductive age in Serbia, where there is no exposure to food fortified with vitamin D. The method of triads was applied. Design: In a cross-sectional study, 422 women completed the Women and Reproductive Health FFQ (WRH-FFQ) during the winter months. From a representative subgroup (n 44), three 24 h dietary recalls and anthropometric parameters were collected as well as a fasting blood sample for vitamin D biomarker analyses. Correlation coefficients were calculated between each of the dietary methods. Validity coefficients, as a correlation between the measured and estimated 'true' exposure, were calculated using the method of triads. Bland-Altman plots were also constructed. Setting: Three major universities in Serbia. Subjects: Healthy young women (n 422) aged 18-35 years. Results: The WRH-FFQ estimate of vitamin D intake for all participants was 4.0 (SD 3.3) mu g/d and 3.1 (SD 2.3) mu g/d for the subgroup. Bland-Altman plots for these intakes showed high agreement. Validity coefficients for the FFQ, 24 h recall and biomarker were. rho(QI) = 0.847 (95 % CI 0.564, 0.928), rho(RI) = 0.810 (95 % CI 0.537, 0.997) and rho(BI) = 0.499 (95 % CI 0.190, 0.840), while the correlation coefficients were 0.686, 0.422 and 0.404. Conclusions: The FFQ applied in the present study is a valid tool for assessing dietary vitamin D intake in women living in Serbia, a region without mandatory vitamin D food fortification.

The interaction of polyphenolic molecules with human serum albumin (HSA) could lead to changes in the reactivity of the HSA Cys34 thiol group (HSA-SH). The influences of enterolactone (EL) and enterodiol (ED) binding on HSA-SH reactivity in fatty acid (FA)-free HSA, and in HSA with bound stearic acid (S) in S/HSA molar ratios of 1 : 1 and 4 : 1, were investigated by the determination of the pseudo first order rate constants (k') for the thiol reaction with 5,5'-dithiobis-(2-nitrobenzoic acid). The binding affinities and binding sites of EL and ED were also determined, using fluorescence measurements of the intrinsic fluorescence of Trp214 and diazepam (binding site marker). EL and ED binding to HSA increased the reactivity of HSA-SH in all assayed HSA-enterolignan complexes by 9.1-33.1%. The strongest effects were obtained for FA-free HSA-enterolignan complexes. S modulated/reduced the effect of EL on HSA-SH reactivity, while its influence on the effect of ED was negligible. The binding of enterolignans to HSA was investigated: the binding constants were the highest for FA-free HSA (EL: 11.64 x 10(4) M-1 and ED: 5.59 x 10(4) M-1 at 37 degrees C) and the lowest for S/HSA 4 : 1-enterolignan complexes (EL: 2.43 x 10(4) M-1 and ED: 1.92 x 10(4) M-1). When the S/HSA ratio was increased, the binding affinities and number of binding sites for EL and ED were decreased. At the same time, a high correlation between binding constants and increased Cys34 reactivity was found (r = 0.974). Competitive experiments using diazepam indicated that the binding of ED and of EL was located in the hydrophobic pocket of site II in HSA. Overall, it is evident that stearic acid could modulate the enterolignan effects on HSA-SH reactivity as well as their binding to HSA. This finding could be important for pharmacokinetics and the expression of enterolignan antioxidant effects in vivo after an intake of lignan rich food.

Objective: The objective of the present study was to examine the external validity of an FFQ designed to estimate dietary vitamin D intake compared with a plasma biomarker and three repeated 24 h dietary recalls in women of reproductive age in Serbia, where there is no exposure to food fortified with vitamin D. The method of triads was applied. Design: In a cross-sectional study, 422 women completed the Women and Reproductive Health FFQ (WRH-FFQ) during the winter months. From a representative subgroup (n 44), three 24 h dietary recalls and anthropometric parameters were collected as well as a fasting blood sample for vitamin D biomarker analyses. Correlation coefficients were calculated between each of the dietary methods. Validity coefficients, as a correlation between the measured and estimated 'true' exposure, were calculated using the method of triads. Bland-Altman plots were also constructed. Setting: Three major universities in Serbia. Subjects: Healthy young women (n 422) aged 18-35 years. Results: The WRH-FFQ estimate of vitamin D intake for all participants was 4.0 (SD 3.3) mu g/d and 3.1 (SD 2.3) mu g/d for the subgroup. Bland-Altman plots for these intakes showed high agreement. Validity coefficients for the FFQ, 24 h recall and biomarker were. rho(QI) = 0.847 (95 % CI 0.564, 0.928), rho(RI) = 0.810 (95 % CI 0.537, 0.997) and rho(BI) = 0.499 (95 % CI 0.190, 0.840), while the correlation coefficients were 0.686, 0.422 and 0.404. Conclusions: The FFQ applied in the present study is a valid tool for assessing dietary vitamin D intake in women living in Serbia, a region without mandatory vitamin D food fortification.

The interaction of polyphenolic molecules with human serum albumin (HSA) could lead to changes in the reactivity of the HSA Cys34 thiol group (HSA-SH). The influences of enterolactone (EL) and enterodiol (ED) binding on HSA-SH reactivity in fatty acid (FA)-free HSA, and in HSA with bound stearic acid (S) in S/HSA molar ratios of 1 : 1 and 4 : 1, were investigated by the determination of the pseudo first order rate constants (k') for the thiol reaction with 5,5'-dithiobis-(2-nitrobenzoic acid). The binding affinities and binding sites of EL and ED were also determined, using fluorescence measurements of the intrinsic fluorescence of Trp214 and diazepam (binding site marker). EL and ED binding to HSA increased the reactivity of HSA-SH in all assayed HSA-enterolignan complexes by 9.1-33.1%. The strongest effects were obtained for FA-free HSA-enterolignan complexes. S modulated/reduced the effect of EL on HSA-SH reactivity, while its influence on the effect of ED was negligible. The binding of enterolignans to HSA was investigated: the binding constants were the highest for FA-free HSA (EL: 11.64 x 10(4) M-1 and ED: 5.59 x 10(4) M-1 at 37 degrees C) and the lowest for S/HSA 4 : 1-enterolignan complexes (EL: 2.43 x 10(4) M-1 and ED: 1.92 x 10(4) M-1). When the S/HSA ratio was increased, the binding affinities and number of binding sites for EL and ED were decreased. At the same time, a high correlation between binding constants and increased Cys34 reactivity was found (r = 0.974). Competitive experiments using diazepam indicated that the binding of ED and of EL was located in the hydrophobic pocket of site II in HSA. Overall, it is evident that stearic acid could modulate the enterolignan effects on HSA-SH reactivity as well as their binding to HSA. This finding could be important for pharmacokinetics and the expression of enterolignan antioxidant effects in vivo after an intake of lignan rich food.

Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states.