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Introduction: Dual antiplatelet therapy (DAPT) of thienopyridines (ticagrelor or clopidogrel) and aspirin is recommended for patients with acute coronary syndromes for a period of 12 months after percutaneous coronary intervention (PCI) and stent implantation. Individual patients` genetic and non-genetic factors may determine bleeding and thrombosis during DAPT. As it was showed already, CYP4F2 rs2108622 may significantly affect antiplatelet treatment with clopidogrel. Our most recent data showed, that CYP4F2 rs3093135 TT variant carriers, users of ticagrelor, had lower platelet aggregation values than non-carriers. Aim: To determine the impact of CYP4F2 rs3093135 TT variant on bleeding and trombosis in patients treated with dual thienopyridine (ticagrelor or clopidogrel) and aspirin antiplatelet therapy after percutaneous coronary intervention (PCI) and stent implantation. Methods: This prospective study was carried out with the patients hospitalized with acute coronary syndromes at the Department of Cardiology of Hospital of Health Sciences, Lithuania between January 2013 till December 2016. All the patients received dual antiplatelet therapy (DAPT) with ticagrelor or clopidogrel and aspirin for at least of 6 months after PCI and stent implantation. From a total of (n=378) patients receiving DAPT, only the patients with CYP4F2 rs3093135 TT variant (n=33) were selected into this study. Bleeding was defined according to Bleeding Academic Research Consortium (BARC) classification. Results: From a total of 33 patients, carriers of CYP4F2 rs3093135 TT variant, 9 (27.3%) patients received ticagrelor and 24 (72.7%) were treated with clopidogrel. Only 2 patients who used ticagrelor had no bleeding events. Seven patients had type 1 or 2 bleeding according to BARC classification and one patient had a major type 3a gastrointestinal bleeding and required a blood tranfusion. The bleeding events were not documented in clop[...].

Kiekvieno vaisto sukeltas tiek farmakologinis (veiksmingumo), tiek toksikologinis poveikis kiekvienam pacientui yra skirtingas, todėl gana dažnai vaistų skyrimas ir vartojimas tampa labai komplikuotas. Prancūzijoje 3.2% hospitalizacijos atvejų yra sąlygoti vaistų. Tai sudaro 320 milijonų eurų sumą per metus. Genetiniai faktoriai, sąlygojantys vaistų farmakokinetiką ir farmakodinamiką, dalinai paaiškina skirtingą vaistų poveikį žmogui. Tyrimo tikslas: 1. Patikrinti galimybę gausinti serume ir plazmoje esančią DNR nauju būdu ir atlikti genetinius tyrimus. 2. Patvirtinti realaus laiko polimerazės grandininės reakcijos (PGR) metodiką aromatazės genui, panaudojus ląsteles, kurių šio geno raiška yra pakankama. Rezultatai: Patikrinta galimybė gausinti serume ir plazmoje esančią DNR, sekvenavus gautą DNR ir sekas palyginus su esančiomis duomenų bazėje : rezultatai teigiami. Nustatyta, kad, ekstrahuojant druskiniu metodu, gaunama daugiau DNR, bet ji blogesnės kokybės, nei naudojant QIAGEN kolonėles. Patvirtinta realaus laiko PGR metodika aromatazės genui, naudojant krūties vėžio ląsteles. There is much variability in the manner individuals respond to drugs, such that the management of some drugs is problematic. In France, the incidence of hospital admissions related to adverse drug reactions is estimated to be 3.2 %, at an annual cost of over 300 millions euros. Genetic factors affecting the pharmacokinetics and pharmacodynamics of drugs partly explain interindividual variability in drug response. Aim of experiment: 1. verify, if it is possible to amplify serum and plasma DNA using new method, and make a genetic research. 2. verify real time polymerase chain reaction to aromatase gene. Find cell line, whish have a sufficient expression of this gene. Results : We verified the possibility to amplify serum and plasma DNA using new method. We made the sequencing of DNA extracts and we compared results in data base : results are positives. It`s important, that QIAGEN extracts are cleanner than salt extracts, but there are few of DNA. We confirmed real time PCR method to aromatase gene, using breast cancer cells.

There is much variability in the manner individuals respond to drugs, such that the management of some drugs is problematic. In France, the incidence of hospital admissions related to adverse drug reactions is estimated to be 3.2 %, at an annual cost of over 300 millions euros. Genetic factors affecting the pharmacokinetics and pharmacodynamics of drugs partly explain interindividual variability in drug response. Aim of experiment: 1. verify, if it is possible to amplify serum and plasma DNA using new method, and make a genetic research. 2. verify real time polymerase chain reaction to aromatase gene. Find cell line, whish have a sufficient expression of this gene. Results : We verified the possibility to amplify serum and plasma DNA using new method. We made the sequencing of DNA extracts and we compared results in data base : results are positives. It`s important, that QIAGEN extracts are cleanner than salt extracts, but there are few of DNA. We confirmed real time PCR method to aromatase gene, using breast cancer cells.

Širdies ir kraujagyslių ligos (ŠKL) siejamos su ateroskleroze, kuri išsivysto dėl uždegiminių procesų kraujagyslėse bei endotelio funkcijos praradimo (disfunkcijos). Naujausių tyrimų duomenimis, antikoaguliantas rivaroksabanas ir širdies nepakankamumo gydymui skiriamas dviejų komponentų sakubitrilo / valsartano derinys greta savo tiesioginių indikacijų pasižymi ir endotelio funkciją gerinančiu poveikiu. Nėra aiškus šių vaistų veikimo mechanizmas. Remiantis ankstesniais Molekulinės kardiologijos laboratorijoje gautais tyrimų rezultatais, padaryta prielaida, jog fermento CYP4F2 aktyvumas gali būti susijęs su minėtų vaistų efektyvumu. Šio bandomojo tyrimo tikslas buvo ištirti rivaroksabano ir sakubitrilo metabolizmą, vertinant HUVEC ir HepG2 ląstelių metabolitus paveikus skirtingomis šių vaistų koncentracijomis bei įvertinti sakubitrilo / valsartano ir CYP4F2 inhibitoriaus poveikį fermento CYP4F2 aktyvumui endotelio ląstelių kultūrose. Tyrimui naudotos žmogaus virkštelės venos endotelio ląstelės (HUVEC) buvo kultivuojamos šešių šulinėlių plokštelėse. Ląstelės buvo paveiktos septyniomis skirtingomis rivaroksabano (0,1 μM, 0,2 μM, 0,5 μM, 1 μM, 2 μM, 5 μM, 10 μM) ir sakubitrilo / valsartano (0,05 μM, 0,1 μM, 0,25 μM, 0,5 μM, 1 μM, 2,5 μM, 5 μM) koncentracijomis. Rivaroksabano tirpalas paruoštas tirpinant vaistą jaučio serumo albumine. Sakubitrilo / valsartano tirpalai buvo paruošti ištirpinus vaistą vandeniniame 0,9 proc. NaCl tirpale. Sakubitrilo metabolitų nustatymas buvo atliktas HUVEC ląstelių lizatuose. Lizatai buvo gauti apdorojant ląsteles lizės buferiu. Ląstelių transfekcija buvo atlikta ląsteles paveikiant miRNR-liposomų kompleksu (1:1). Praėjus 24 valandoms, buvo paveikiama vaistu. Transfekcija hsa-miR-24-3p imitatoriumi buvo atlikta siekiant slopinti CYP4F2 geno, koduojančio CYP4F2 fermentą, raišką ir įvertinti šio fermento poveikį sakubitrilo metabolizmui. Metabolitai surinktose ląstelių terpėse buvo nustatyti naudojant skysčių chromatografijos-kvadrupolio praskriejimo laiko masių spektrometrijos (UPLC-QTOF-MS) metodą. Kiekybinis susidariusių junginių, esančių ląstelių terpėse ir HUVEC ląstelių lizatuose, įvertinimas buvo atliktas naudojant ultraefektyviosios skysčių chromatografijos-trigubo kvadrupolio masių spektrometrijos (UPLC-TQD-MS) metodą. Tyrimo metu buvo identifikuoti rivaroksabano M-1, M-2, M-5, M-8, M-10, M 11, M-18 metabolitai ir sakubitrilo / valsartano M1 metabolitas HUVEC ląstelėse paveiktose skirtingomis vaistų koncentracijomis. Nustatytas naujas junginys (Z)-dokos-13-enamidas, kurio koncentracija kito priklausomai nuo rivaroksabano koncentracijos. Gauti duomenys parodė, kad sakubitrilas nėra pilnai metabolizuojamas į aktyvųjį metabolitą (sakubitrilatą) M1, kai HUVEC ląstelės yra veikiamos didesne negu 0,1 μM vaisto koncentracija. CYP4F2 fermento koncentracija padidėjo 2,6 karto, HUVEC ląsteles paveikus 5 μM sakubitrilo / valsartano koncentracija, o CYP4F2 baltymo koncentracija HepG2 ląstelėse mažėjo didėjant vaisto koncentracijai. Hsa-miR-24-3p imitatorius slopino sakubitrilo biotransformaciją į aktyvųjį metabolitą M1 HUVEC ląstelėse., Cardiovascular disease (CVD) is associated with atherosclerosis, which develops due to inflammatory processes in the blood vessels and loss of endothelial function (dysfunction). Recent studies have shown that anticoagulant rivaroxaban and two-component combination of sacubitril / valsartan used for the treatment of heart failure may also have an endothelial-affecting activity in addition to their direct indications. The mechanism of action of these drugs is still not clear. Based on previous research carried out in the Laboratory of Molecular Cardiology, it has been suggested that the activity of CYP4F2 enzyme may be related to the efficacy of these drugs. The aim of this pilot study was to investigate the metabolism of rivaroxaban and sacubitril by evaluating the metabolites in HUVEC and HepG2 cells treated with different concentrations of drugs and to evaluate the effect of sacubitril / valsartan and inhibitor of CYP4F2 on the activity of CYP4F2 enzyme in endothelial cell cultures. The human umbilical vein endothelial cells (HUVEC) used in this study were cultivated in a six-well plate. Cells were treated with seven different concentrations of rivaroxaban (0,1 μM, 0,2 μM, 0,5 μM, 1 μM, 2 μM, 5 μM, 10 μM) and sacubitril / valsartan (0,05 μM, 0,1 μM, 0,25 μM, 0,5 μM, 1 μM, 2,5 μM, 5 μM). Rivaroxaban solution was prepared by dissolving the drug in bovine serum albumin. Sacubitril / valsartan solutions were obtained by dissolving the drug in aqueous 0.9 % NaCl solution. Screening for sacubitril metabolites was performed in HUVEC cell lysates. Lysates were obtained by treating cells with lysis buffer. Cell transfection was performed by treating the cells with a miRNA-liposome complex (1:1). Cells were treated with the drug after 24 hours. Transfection with hsa-miR-24-3p mimic was performed to suppress the expression of CYP4F2 gene encoding the CYP4F2 enzyme and to evaluate the effect of this enzyme on sacubitril metabolism. Metabolites in the collected cell media were identified using liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) method. Quantification of the formed compounds of in cell media and HUVEC cell lysates was performed by ultra-efficient liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQD-MS) technique. Metabolites M-1, M-2, M-5, M-8, M-10, M 11, M-18 of rivaroxaban and the M1 metabolite of sacubitril / valsartan have been identified in HUVEC cells exposed to different drugs concentrations. A new compound (Z)-doco-13-enamide was identified, which concentration varied with the concentration of rivaroxaban. The obtained data showed that sacubitril is not completely metabolized to the active metabolite M1 (sacubitrilat) when HUVECs are exposed to concentrations greater than 0.1 μM. CYP4F2 enzyme concentrations increase 2.6-fold with 5 μM sacubitril / valsartan treatment in HUVECs, and CYP4F2 protein concentration decreased in HepG2 cells with increasing drug concentrations. The hsa-miR-24-3p mimic inhibited the biotransformation of sacubitril to the active metabolite M1 in HUVECs.

The aim of this study – to determine and evaluate changes in the expression of potential target genes and the concentration of CYP4F2 protein in endothelial cell cultures after exposure to a new generation anticoagulant rivaroxaban and hsa-miR-24-3p mimic. Therefore, five study objectives were defined: 1. To determine changes in the expression of CYP4F2, HNF4α in endothelial cells after treatment with different concentrations of rivaroxaban. 2. To determine changes in the expression of CYP4F2, HNF4α in endothelial cells after treatment with hsa-miR-24-3p mimic. 3. To determine the concentration of CYP4F2 protein in endothelial cells after treatment with different concentrations of rivaroxaban. 4. To determine the concentration of CYP4F2 protein in endothelial cells after treatment with hsa-miR-24-3p mimic. 5. To determine changes in the expression of CYP4F2, HNF4α in endothelial cells after induction of HNF4α expression by glucocorticoid dexamethasone. The object of this study – changes in gene expression and concentration of CYP4F2 protein in vitro after exposure to rivaroxaban and hsa-miR-24-3p. Commercial human umbilical vein endothelial cell culture (HUVEC) was selected as a suitable model to study the selected object of research. In this study, endothelial cells were treated with different concentrations of rivaroxaban. Forward transfection with hsa-miR-24-3p mimic was also performed. Total RNA was extracted from prepared cell suspensions performing phenol/chloroform and glass fiber filter-based RNA purification. Complementary DNA was synthesized using a commercial kit. The expression level of target genes was determined by quantitative real-time polymerase chain reaction method using SYBR Green fluorescent dye. Relative expression results were calculated using ΔΔCt method. Concentration of CYP4F2 protein in cell lysates was determined using a commercial ELISA kit. Statistical analysis was performed using both IBM SPSS Statistics v.20.0 and GraphPad Prism v.6 software. Results showed that after treatment with rivaroxaban the expression of CYP4F2 gene in HUVECs decreased in (rivaroxaban) concentration-dependent manner as compared to control cells: treatments with 0.1, 0.2, 0.5, 1, 2, 5 and 10 μM rivaroxaban downregulated the expression of CYP4F2 by 1.44 (p = 0.046), 1.71 (p = 0.026), 1.89 (p = 0.028), 1.93 (p = 0.013), 2.05 (p = 0.004), 2.4 (p = 0.009), 2.72-fold (p = 0.009), respectively. The expression of HNF4α gene in treated cells was not detected. After transfection with hsa-miR-24-3p mimic the expression of CYP4F2 gene in HUVEC`s resulted in a 1.7-fold decrease (p = 0.002). The expression of HNF4α gene was not detected. After treatment with rivaroxaban the concentration of CYP4F2 protein in treated cells did not alter significantly. Transfection with hsa-miR-24-3p mimic resulted in a 6.4-fold decrease of CYP4F2 protein levels (p = 0.029) compared to negative control cells. After treatment with dexamethasone in order to induce HNF4α gene expression, the expression of HNF4α in treated cells was not detected. After treatment with 1 μM dexamethasone the expression of CYP4F2 gene in HUVECs resulted in a 1.96-fold decrease (p = 0.029) compared to control cells. In conclusion, the expression of CYP4F2 in HUVECs decreased after treatment with rivaroxaban (in a concentration-dependent manner), hsa-miR-24-3p mimic and dexamethasone. The concentration of CYP4F2 protein decreased due to an increase of hsa-miR-24-3p in endothelial cells. The expression of transcription factor HNF4α in endothelial cells was not determined.

Final Master‘s thesis by G. Jablonskytė, title „The cell culture research: metabolism of new generation anticoagulants“. Supervisor V. Tatarūnas; Laboratory of Molecular Cardiology at the Institute of Cardiology, Lithuanian University of Health Sciences. - Kaunas. The aim of the study: to determine the metabolites of rivaroxaban and new compounds, involved in the effect of rivaroxaban, and to investigate the influence of inhibitors of CYP4F2 enzyme on the metabolism of anticoagulant in endothelial cell cultures. Objectives: 1. To identify metabolites of rivaroxaban in the media of endothelial cells after the exposure of HUVECs to different drug concentrations. 2. To determine new compounds that are formed upon the exposure of HUVEC cells to different concentrations of rivaroxaban. 3. To determine the effect of different concentrations of rivaroxaban and inhibitors of CYP4F2 enzyme on the levels of CYP4F2 enzyme in endothelial cell lysates. 4. To determine CYP4F2 gene variants in endothelial cells. Object: commercial human umbilical vein endothelial cells (HUVECs). Methods: cell cultivation in flasks, genotyping by real-time polymerase chain reaction, methods of liquid chromatography and mass spectrometry, enzyme-linked immunosorbent assay ELISA. Results and conclusions: rivaroxaban and M-1, M-2, M-5, M-8, M-10, M-11, M-18 metabolites were identified from cultured HUVEC cells after exposure with rivaroxaban. In addition, it was determined that the concentration of a new compound, which previously has not been linked with the effect of rivaroxaban, decreased as the concentration of rivaroxaban increases. CYP4F2 levels decreased with increasing rivaroxaban concentrations. Pre-treatement with 0.5 μM of rivaroxaban solution decreased the levels of enzyme in cell cultures by 17.2 %, 1 μM rivaroxaban - by 81.3 %, and 2 μM of rivaroxaban by 85.5 %. Treatment with Hsa miR 24-3p reduced the levels of CYP4F2. Enzyme levels decreased by 11.9 % in HUVECs exposed to 120 nM hsa-miR-24-3p. The CYP4F2 rs2108622 C/C, rs1558139 G/G, rs2074902 T/T and the CYP4F2 rs3093135 A/T variants were detected in HUVEC`s cells.

Author: Rita Norvilaitė Title: The impact of clinical and genetic factors on platelet aggregation during clopidogrel or ticagrelor antiplatelet therapy in patients with acute coronary syndromes. The aim: To evaluate the impact of clinical and genetic factors on platelet aggregation (PA) during clopidogrel or ticagrelor antiplatelet therapy in patients with acute coronary syndromes. Objectives: To evaluate the impact of clinical and genetic factors on platelet aggregation of clopidogrel or ticagrelor in patients with acute coronary syndromes: 1) during induction of antiplatelet therapy and 2) after 4 weeks of antiplatelet therapy; 3) To compare the impact of clinical and genetic factors on platelet aggregation of clopidogrel or ticagrelor during induction and after 4 weeks of DAPT. Methods: Platelet aggregation, genotyping and analysis of arachidonic acid metabolites in blood samples were performed for all studied patients. Participants: Totally 467 patients with ACS after PCI and stent implantation during induction of DAPT were included in the study. Results: Lower PA values were determined in male patients users of clopidogrel or ticagrelor during induction of antiplatelet therapy after stent implantation. Higher PA values were determined in clopidogrel treated patients with diabetes during induction and in ticagrelor treated patients of 65 years old. Significantly lower PA values were observed in smoker users of clopidogrel. Concomitant amiodarone or metformin use had a significant impact on antiplatelet therapy in clopidogrel or ticagrelor users during induction of antiplatelet therapy. Significantly higher PA values were determined in concomitant ticagrelor and ACE inhibitor users or in clopidogrel and glimepiride users after 4 weeks of induction. The carriers of CYP2C19*1*2 variant who were prescribed clopidogrel or ticagrelor had significantly higher PA values than CYP2C19*1*1 variant carriers during induction and after 4 weeks of induction of antiplatelet therapy. Higher 6-trans-leukotriene B4 concentration was observed in clopidogrel but not in ticagrelor treated patients after 4 weeks of treatment. Conclusions: Patient gender, diabetes, smoking and concomitant use of metformin, amiodarone, ACE inhibitor, and glimepiride have a significant impact on the therapeutic effect of clopidogrel and ticagrelor antiplatelet therapy. The CYP2C19 variant has a significant effect on platelet reactivity during induction and after 4 weeks in clopidogrel or ticagrelor – treated patients. Regardless of CYP2C19, lower PA was determined in ticagrelor users than in clopidogrel users. Key words: clopidogrel metabolism, ticagrelor pharmacogenomics, CYP2C19.

Valproic acid (VPA) is one of the most widely used antiepileptic drugs in the world. It is also one of the most frequently used to treat epilepsy in children. Valproic acid dosages required to achieve therapeutic effect vary among patients. Therapeutic drug monitoring is usually used to monitor valproic acids plasma levels, however, therapeutic effect and the risk of side effects show a poor correlation with the concentration of valproic acid. The aim of this study was to determine the effect of clinical factors, genetic variations in UGT1A6 and CYP2C9 genes (and their frequencies) and valproic acid and it`s metabolite concentrations on the therapeutic dosage of valproic acid in children with epilepsy. In total n = 51 children (age median 9.5, range from 3 to 17 years of age) who were prescribed treatment with valproic acid in the Department of Neurology of the Hospital of Lithuanian University of Health Sciences „Kauno Klinikos“, were included in this study. Sanger sequencing was performed to determine genetic variations in UGT1A6. Real time PCR was used to determine genetic variations in CYP2C9. Patient blood plasma concentrations of valproic acid and it`s metabolites were determined with liquid chromatography - mass spectrometry. Results: Older patients and male patients had higher concentrations of valproic acid-glucuronide. Genetic changes in UGT1A6 and CYP2C9 had no effect on plasma levels of valproic acid and it`s metabolites. According to the results of this study, it is recommended to use a larger number of patients for future research: it may reveal the impact of CYP2C9 genetic changes on the metabolism of valproic acid. Also, the function of the kidneys should be evaluated to better determine the effects of age on valproic acid metabolism and therapeutic dosage.

The aim: The aim of this research is to set how (I) type of plasminogen activator inhibitor do the influence to the blood coagulation system activity. The objectives of the study 1) To identify PAI-1 4G / 5G genotype effect for the blood coagulation system activity, according to platelet aggregation, exposured by ADP, ADR indicators and INR. 2) To identify the total PAI-1 4G / 5G genotype and adjacent used antiaggregant aspirin effect on the blood coagulation system activity, according to platelets aggregation, exposured with ADP, ADR indicators and TNS. 3) To evaluate examined patients PAI-1 4G / 5G genotype frequency in men and women. Research methodology: During the test we examined the patients who after valve surgery were being treated with atiagreggants and anticoagulation drugs. The aim of our study was to evaluate the association of clinical and genetic factors with coronary artery occlusion in patients with myocardial infarction.All patients had the PAI-1 4G / 5G -675 (rs1799889) polymorphisms detection was achieved by using Sanger sequencing in a sample. Platelet aggregation was evaluated after exposure aggregation inducers of ADP and ADR. Evaluated the international normalized ratio - INR. Results: We found new biomarkers that have a significant impact on the blood coagulation system activity in patients who were in treatment with anticoagulants and antiplatelets after heart valve surgery. PAI-1 had a statistically significant effect on INR. Patients with 5G / 5G, had a higher INR 2.7 ± 1.29 compared with patients having a 4G / 5G INR 1.70 ± 0.69 (p = 0.003). However we determined a tendency (p = 0, 07) that patients with the 4G / 4G also had lower INR 1, 90 ± 0, 61 comparing with 5G / 5G. Conclusion: The results showed that higher INR indicators had 5G / 5G patients than the 4G / 5G or 4G / 4G patients. In addition used aspirin in this study had no significant effect to the blood coagulation system activity. The frequency of PAI-1 4G / 5G genotype in men and women was not significantly different.

The aim of the study: To show the influence of clinical and genetic factors on warfarin dosage during induction and long-term treatment in patients who followed heart valve surgery. The objectives of the study: 1. To determine the impact of amiodarone usage on warfarin dosage after heart valve surgery during induction and long-term treatment. 2. To evaluate the influence of CYPC9 and VKORC1 genetic polymorphism on warfarin dosage after heart valve surgery durint induction and long-term treatment. 3. To determine the impact of CYP4F2 rs1558139 gene polymorphism on warfarin dosage after heart valve surgery durint induction and long-term treatment. Objects ant methods: This study involved 91 patient who reached stable warfarin dosage (the therapeutic range of INR (from 2 to 3.5) was achieved and maintained for two days of treatment) during discharge from the hospital. Totally 72 patients, who used warfarin for at least 3 months, were included into the further study. Study involved 38 (52.8 %) men and 34 (47.2 %) women. DNA extraction, DNA quality assessment and real-time polymerase chain reaction (RT-PCR) were performed at the laboratory of Molecular Cardiology of LUHS.Prevalence of genotypes is presented in % (percent). Warfarin dosage is presented as mean ±SD. A p value

The main problem of this work – the expected antiplatelet effect is not always achieved and it depends on the patients` genotype. In order to achieve the required therapeutic effect, the genes, affecting the therapeutic effect of these drugs should be performed The aim of this study was to estimate the factors, impacting the resistance to antiplatelet therapy in patients following stent implantation (SI) or in patients after coronary artery bypass grafting (CABG) by using platelet aggregation, clinical and genetic data. Main tasks: to estimate the influence of clinical factors on the platelet aggregation (PA) during induction and long-term treatment in patients following SI, also, to find the impact of genotype and concomitant use of drugs on PA values in these patients; to find the distribution of genotypes in the patients who followed antiplatelet therapy after CABG or SI. Methods: Real-Time PCR was used to determine the CYP2C19*2 and CYP4F2 G1347A genotypes. Fisher exact test was used to compare categorical variables. Hardy Weinberg method was used to show the concordance of analyzed patients genotypes to a normal population genotype distribution. A p value

Objective: The purpose of this study was to assess platelet aggregation (PA) in the diabetic and in non-diabetic patients, also, to determine the impact of anthropometric, demographic factors and concomitant drug use on the PA in the presented groups of the patients. Methods: The study sample consisted of the (n=286) patients who were hospitalized due to ACS and were prescribed DAPT (aspirin and clopidogrel) at the Department of Cardiology of the hospital of LUHS Kauno Klinikos. Patients followed DAPT for at least 2 weeks from hospitalization. PA was assessed by using light transmittance aggregometry. Adenosine diphosphate (ADP), epinephrine (ADR), and arachidonic acid (ARA) were used to induce PA. The results were presented as the mean ± standard deviation (SD). The results were considered as statistically significant when p≤0.05. Results: PA induced by ADP (26.12 ± 7.76 %agr vs. 35.57 ± 14.11 %agr) and ADR (29.67 ± 12.73 %agr vs. 35.99 ± 17.08 %agr) was significantly higher in the diabetic patients. PA with ARA levels did not differ significantly in the presented patient groups. Significantly lower PA values were determined in the non-diabetic patients vs diabetic patients: younger than 60 years old (26.71 ± 8.01 %agr vs. 33.05 ± 13.72 %agr), older than 60 years of age (25.69 ± 7.58 %agr vs. 36.38 ± 14.25 %agr), males (25.47 ± 7.93 %agr vs. 35.12 ± 14.41 %agr) and females (28.02 ± 7.00 %agr vs. 36.45 ± 13.72 %agr), in the patients, according to their waist circumference (99 cm: 26.94 ± 8.71 %agr vs. 35.77 ± 14.38 %agr. PA was significantly higher in the diabetic patients compared to nondiabetic patients, regardless to the concomitant drug usage: ACE inhibitors, aspirin, furosemide/torasemide, ISMN, ivabradine, ranitidine, spironolactone or statins. Conclusions: DAPT with aspirin and clopidogrel was less effective in the diabetic patients’ sample. Diabetic patients had significantly higher PA levels than compared to the non-diabetic patients despite their age, gender, waist circumference or concomitant drug usage.

Objective and tasks. To evaluate and to determine whether CYP2C9 *2, *3, VKORC1 *2, *3 and CYP4F2 G1347A genotypes and clinical factors contribute to warfarin dosage in patients after heart valve surgeries. Methods. During the initiation of warfarin therapy, 175 patients, who had heart valve surgeries, were included into the study. During the long term treatment, totally 113 patients were analyzed. Patient anthropometric, clinical data (biochemical tests, used medications, addictions) and patient INR were obtained from the case histories. Genomic DNA was extracted from peripheral blood. The real – time polymerase chain reaction was used to detect the CYP2C9 * 2 (rs1799853) and *3 (rs1057910), VKORC1 *2 (rs9934438), VKORC1 *3 (rs7294) and CYP4F2 G1347A (rs2108622) genotypes. Statistical analysis. Results are presented as a mean and standard deviation (mean ± SD). A p≤0,05 was taken as statistically significant. Frequency of genotypes and alleles in patient’s sample is presented as % (percent). Results. The patients, users of amiodarone, required lower dosages of warfarin during treatment initiation, as compared to the non-users. During the long – term treatment period, the users of torasemide, calcium channel blockers and benzodiazepines, required lower warfarin dosages as compared to non-users of these drugs. During the long term treatment, patients with elevated creatinine or elevated alkaline phosphatase activity, required lower dosages of warfarin as compared to the patients with normal levels of these enzymes. Patients with such addictions (smoking and alcohol use) required higher dosages of warfarin. Besides the clinical factors, warfarin dosage was also influenced by these genetic variants (both at initiation of the warfarin therapy and during the long term treatment): CYP2C9*1*3 heterozygotes used lower warfarin dosages as compared to the wild-type homozygotes, VKORC1 C1173C carriers required higher dosages of warfarin than comparing to C/T and T/T genotype carriers, VKORC1 G3730G carriers required lower dosages of warfarin compared to A/A genotype carriers. The carriers of CYP4F2 A1347A required higher warfarin dosages, as compared to CYP4F2 G/G cariers, but only at initiation of warfarin therapy. Conclusions: Dosage of warfarin is influenced by such a clinical factors as the activity of plasma serum creatinine, the activity of alkaline phosphatase, medications - amiodarone, torasemide, calcium channel blockers, benzodiazepines, patient addictions (smoking, alcohol consumption). The genotypes CYP2C9 * 1 * 3, VKORC1 C1173T, VKORC1 G3730A and CYP4F2 G1347A are also significant in determining warfarin dosage.

Siauro terapinio indekso vaistų vartojimas ir farmakokinetikos tyrimų interpretavimas reikalauja specialių farmakokinetikos žinių. Mokslinės literatūros šaltiniuose teigiama, jog gydymo procese dalyvaujant klinikiniams vaistininkams vaistų vartojimas yra optimizuojamas. Lietuvoje panašios vaistininkų veiklos nėra, todėl tyrimo tikslas buvo įvertinti siauro terapinio indekso vaistų vartojimo racionalumą ir nustatyti klinikinės farmacijos paslaugos poreikį stacionarinio gydymo įstaigoje. Darbo uždaviniai: nustatyti ir įvertinti farmakoterapinių problemų, siejamų su siauro terapinio indekso vaistų (vankomicino, ciklosporino, digoksino, gentamicino) vartojimu, paplitimą tretinio lygio ligoninėje; kiekybiškai ir kokybiškai įvertinti nustatytų farmakoterapinių problemų rizikos veiksnius; įvertinti vankomicino farmakokinetikos stebėsenos praktiką ligoninės skyriuose; įvertinti farmacininko konsultacijos įtaką vankomicino farmakokinetikos stebėsenai. Tyrimui pasirinktų siauro terapinio indekso vaistų farmakokinetikos tyrimų analizė atskleidė klinikinės farmacijos paslaugos poreikį optimizuojant tokių vaistų vartojimą Lietuvos ligoninėse. Nustatytų farmakoterapinių problemų rizikos veiksnių analizė identifikavo praktikoje dar visuotinai nepripažintą padidintą inkstų klirensą ir jo reikšmingą įtaką vaistų veiksmingumui. The prescription of narrow therapeutic index drugs (NTID) and interpretation of pharmacokinetic measurements require special pharmacokinetic knowledge. Scientific publications report that active participation of clinical pharmacists in the treatment process results in optimization of the drugs use. In Lithuania, there are no similar services provided by pharmacists up till now. Aim of the study: to evaluate the rationality of the use of NTIDs and to determine the demand for clinical pharmacy services in hospitals. Objectives of the study: to identify and evaluate the prevalence of drug-related problems associated with the use of NTIDs (vancomycin, cyclosporine, digoxin, and gentamicin) in the tertiary-level hospital; to perform quantitative and qualitative analysis of risk factors for the identified drug-related problems; to evaluate the practice of pharmacokinetic vancomycin monitoring in hospital departments; to evaluate the influence of pharmacist intervention on pharma¬cokinetic vancomycin monitoring. The analysis of pharmacokinetic measurements of NTIDs selected for this study revealed the demand for clinical pharmacy services while optimizing NTIDs use in the Lithuanian hospitals. The analysis of risk factors associated with the identified drug-related problems identified augmented renal clearance, which is not widely acknowledged in practice, and its significant impact on the effectiveness of drugs.