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ABSTRACT: The aim of this study was to determine the association of bleeding as a complication of warfarin therapy with polymorphism of CYP2C9 gene (alleles 1, 2 and 3). The CYP2C9 is the main enzyme for warfarin metabolism. Study included 181 patients receiving warfarin for at least one month. Allele 1 of CYP2C9 gene (in 94.5%) and genotype *1/*1 (57.5%) prevailed. Allele 3 was found in 12.7% patients. Bleeding side-effects occurred in 18 patients (10%). Patients with allele *1 needed significantly higher maintenance warfarin dose (p=0.011). Those with allele *3 had significantly lower maintenance warfarin dose (p=0.005) and higher prothrombin time (PT) at induction (p=0.034). Bleeding occurred significantly more often in those with lower maintenance warfarin dose (p=0.017). Patients with allele *3 had increased risk of bleeding, with marginal significance (p=0.05). Polymorphism of CYP2C9 could determine dose of warfarin therapy and thus it could be related to the risk of bleeding complications. Allele *3 carriers need lower warfarin dose. Therefore, initially reduced warfarin induction dose in allele *3 carriers could avoid more prolonged PT and decrease the risk of bleeding complication.
Collegium antropologicum 07/2008; 32(2):557-64. · 0.61 Impact Factor
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ABSTRACT: The aim of the study was to investigate the association between methylenetetrahydrofolate (MTHFR) genotypes and levels of homocysteine (Hcy), folate, vitamin B12 and lipids as well as the association between apolipoprotein E (apo E) genotypes and levels of lipids in a Croatian healthy control group and a group of patients with > 70% carotid stenosis (CS). The study included 98 Croats, 38 patients with > 70% carotid stenosis and 60 age- and sex-matched controls. The MTHFR and apo E genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), Hcy by enzyme immunoassay, vitamins by immunochemiluminiscence, and lipids by spectrophotometric method. There was no difference between control subjects and CS patients in the distribution of C677T MTHFR genotypes (p=0. 786) and alleles (p=0.904), however, differences in the frequencies of apo E genotypes (p=0.012) and alleles (p=0.029) were statistically significant. The odds ratio for apo E 3/4 genotype was 3.93 (95% CI 1.23-12.61). Hyperhomocysteinemia (> or =15 micromol/L) was found in 11% of CS patients and 5% of control subjects. Total cholesterol, triglycerides, vitamin B12 and folate were statistically different in "all MTHFR genotypes" (p<0.001, p<0.01, p=0.044 and p=0.036, respectively), and in TC/TT (p<0.001, p=0.003, p=0.030 and p=0.032, respectively) groups. The levels of total cholesterol, LDL cholesterol and triglycerides in the apo E 3/3, and total cholesterol in the apo E 3/4 group yielded statistical difference. An association was found of apo E 3/4 genotype but not of MTHFR genotypes with the risk of CS. MTHFR and apo E affect blood lipid levels, which was statistically confirmed. An association was also recorded between hyperhomocysteinemia and patients with CS. Vitamin status in CS showed a statistically verified association with TC/TT MTHFR genotype. In the group of patients with TC/TT MTHFR genotype, lower vitamin B12 and higher folate values were recorded. The results of multiple logistic analysis showed that there was no statistical significance of Hcy levels (OR 2.403, p=0.334) or conventional vascular risk factors such as smoking habit (OR 0.505, p=0.149), age (OR 1.048, p=0.087) or sex (OR 2.037, p=0.112) in predicting CS.
Collegium antropologicum 01/2007; 30(4):871-8. · 0.61 Impact Factor
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Biochemia Medica 01/2007; 17(2):242-246. · 1.34 Impact Factor
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ABSTRACT: In this study, we investigated the role of glutathione S-transferase P1 (GSTP1) polymorphisms in the pathogenesis of Alzheimer's disease (AD). We genotyped the GSTP1 polymorphisms in exon 5 (A313G) and exon 6 (C341T) by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 56 Croatian patients with AD and 231 controls. Distributions and frequencies of GSTP1 genetic variants were not statistically different between AD patients and healthy controls. Higher frequencies of the mutant genotypes were observed in AD patients (13% for both A313G and C341T) when compared with control subjects (7% for A313G and 8% for C341T), but association of GSTP1 GG (OR 2.057, 95% CI 0.796-5.315, p=0.094) and TT (OR 1.691, 95% CI 0.669-4.270, p=0.514) genotypes with an increased risk of AD was not confirmed by statistical analysis. The frequencies of GSTP1 alleles (A, B, C, D) did not significantly differ between AD patients and controls and they were indicated as follows: 52.7%, 15.2%, 12.5% and 19.6% for AD cases and 58.4%, 14.1%, 14.1% and 13.4% for controls. The estimation of the GSTP1 haplotype distribution showed that GSTP1*A/GSTP1*B and GSTP1*A/GSTP1*C haplotypes were less frequent, while GSTP1*B/GSTP1*B and GSTP1*C/GSTP1*D haplotypes were more frequent in AD patients than in controls. In conclusion, the involvement of GSTP1 alleles in individual susceptibility to AD was not confirmed as statistically significant in the tested Croatian Caucasian population. A possible role of GSTP1 in the complex etiopathogenesis of AD is further discussed, based on observed differences in haplotype distribution and higher frequencies of mutant genotypes in AD patients.
Clinical Chemistry and Laboratory Medicine 04/2004; 42(3):334-9. · 2.15 Impact Factor
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ABSTRACT: CYP2C9, an isoform of the cytochrome P450 enzyme, is involved in the metabolism of most of the drugs of choice for the treatment of thromboembolic disorders. Functional polymorphism is associated with two variant alleles (alleles *2 and *3) encoding CYP2C9 enzymes with a potentially different catalytic activity. The aim of the study was to determine the frequency of the CYP2C9 polymorphism in a representative sample of the Croatian population (n = 177) and to assess the association between the CYP2C9 polymorphism and the warfarin dose in patients with thromboembolism (n = 181). The CYP2C9 genotype was determined by polymerase chain reaction-restriction fragment length poymorphism (PCR-RFLP). According to the CYP2C9 genotype distribution, 31.2% of the healthy subjects were identified with a heterozygous or homozygous CYP2C9 variant genotype. The frequency of 2C9*2 and 2C9*3 alleles was 12.4% and 3.7%, respectively. There was no gender-related genotype or allele difference. In thromboembolism patients, the frequency of CYP2C9 alleles *2 and *3 was 17.4% and 6.6%, respectively, and did not differ significantly from the control group. Almost half (42.5%) of the patients carried at least one variant CYP2C9 genotype. The allele difference between patient subgroups receiving warfarin doses lower and higher than the optimal warfarin dose (4.1 mg/day) was significant (p = 0.027), especially for allele 2C9*3 (p = 0.019; OR 3.250, 95%, CI 1.263-8.413). Comparison of the warfarin dose between patients with different genotypes yielded a significant dose difference between the patients with wild-type genotype and those with variant genotypes (Kruskall-Wallis, chi2 = 9.745, p = 0.008). The results of the association of each of five genotype combinations with the warfarin maintenance dose revealed it to be significantly related to the genotype (Kruskall-Wallis, chi2 = 12.854, p = 0.025). Expressed as percentage of the mean dose in patients with wild-type alleles, the mean warfarin maintenance dose was 92% in 2C9*2 heterozygotes, 74% in 2C*3 heterozygotes, 61% in 2C9*2 homozygotes, 34% in 2C9*3 homozygotes and 63% in compound heterozygotes for 2C9*2 and 2C9*3. Although the mean maintenance dose in homozygous *2/*2 and compound *2/*3 genotype patients was markedly lower (mean 2.66 mg and 2.75 mg, respectively, vs. 4.37 mg), statistical analysis yielded no significance because of the small number of patients carrying these genotypes. A significantly lower maintenance dose was observed in the subgroup of heterozygous *1/*3 genotype patients (p = 0.022). These preliminary results suggest a significant association of the CYP2C9 polymorphism with the warfarin dose and underline the importance of pre-therapeutic genotyping to identify the subjects likely to develop undesirable drug effects.
Clinical Chemistry and Laboratory Medicine 02/2004; 42(1):72-8. · 2.15 Impact Factor
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ABSTRACT: The diagnosis of acute myocardial infarction (AMI) and unstable angina is based on typical clinical signs, electrocardiogram (ECG) changes, and serial measurements of characteristic serum enzymes, especially troponins. Today, newer biochemical markers are used in assessing these conditions, as well as minor myocardial damage (MMD).
The aim of this study was to determine the existence of MMD by the analysis of new biochemical markers including cardiac troponin T (c-TnT), troponin I (c-TnI), CK-MB activity (CK-MBact), CK-MB mass (CK-MBmass), a conventional marker of total creatine kinase (CK), and 12-lead ECG monitoring in two groups of critically ill patients admitted to the intensive care unit (ICU). Group 1 (n=52) consisted of the patients with heart failure, unexplainable hypotension, chronic obstructive pulmonary disease in acute exacerbation, acute severe pancreatitis, sepsis, pulmonary embolism, diabetes with complications, liver cirrhosis, and tachycardia >120/min who suffered chest discomfort without evident ECG signs of AMI. Group 2 consisted of patients (n=73) with acute gastrointestinal bleeding, poisoning, and miscellaneous conditions without chest discomfort or ECG signs of AMI.
Of the 52 critically ill patients, 21.2% were positive for troponin T and 11.5% for troponin I. Group 1 patients showed a prevalence of elevated total CK (28.8%), CK-MBact (7.7%), and CK-MBmass (32.7%). In group 2, positive troponin I was found in 19.7% of patients, troponin T was negative (0.00%), with elevated total CK (23.9%), CKMBact (7.0%), and CK-MBmass (2.3%). The mean concentrations of c-TnT, c-TnI, total CK, CK-MBact, CK-MBmass were higher in group 1 than in group 2, however, without statistical significance. The best positive correlation was recorded between CK-MBact and CK-MBmass (r=0.63). The c-TnI showed best discrimination and accuracy in the assessment of MMD under the ROC curves surface (0.84), while the accuracy was low for all other markers analyzed. Discrimination and accuracy were moderate for all markers analyzed.
CK-MBact (92.3%) showed the highest specificity, followed by c-TnI (88.5%) and c-TnT (75.6%). The sensitivity was low for all markers analyzed. Concerning specificity, CK-MBact proved to be the best biologic marker for the assessment of MMD, followed by c-TnI and c-TnT. Correct clinical assessment according to marker accuracy and discrimination can be achieved by use of c-TnI, however, with a moderate degree of accuracy and discrimination for the detection of MMD in critically ill patients admitted to ICU.
Acta medica Croatica: c̆asopis Hravatske akademije medicinskih znanosti 01/2004; 58(5):381-8.
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ABSTRACT: The aim of this study was to investigate the frequency of C677T methylenetetrahydrofolate reductase (MTHFR) mutation in healthy Croatian volunteers and in patients with atherosclerosis.
The C677T MTHFR gene mutation was determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 640 subjects, residents of the Zagreb city or Zagreb surroundings. Control group (n=298) was healthy blood donors. Patients (n=342) were divided into two groups of those with coronary heart disease, CAD (n=247) and those with >60% carotid stenosis, CS (n=95).
CC genotype was recorded in 45% of healthy volunteers and 46% of patients (46.3% with CS and 46.2% with CAD). TC genotype was found in 49% of healthy volunteers and 45% of patients (46.3% with CS and 44.9% with CAD). There was no significant difference (p>0.05) from the control group in the genotype or allele frequency either for the overall group of patients with atherosclerosis or for the patient subgroups.
The preliminary study of MTHFR polymorphism in control subjects and cardiovascular disease/carotid stenosis patients revealed that in Croats there was a low frequency of TT genotype (6% in controls vs. 9% in patients) and T allele (31% for cases and controls). Additionally, our results did not show significantly higher frequency of MTHFR mutation in CAD and CS studied groups.
Clinica Chimica Acta 09/2003; 335(1-2):95-100. · 2.54 Impact Factor
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Biochemia Medica (biochemia-medica@hdmb.hr); Vol.16 No.1.
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ABSTRACT: Introduction: A case of venous thromboembolism (VTE) in a 14-year-old boy who developed deep venous thrombosis of the right leg followed by pulmonary embolism complicated with pulmonary infarction is described in this article. Aim: To present results of laboratory diagnosis suggesting the possible cause of VTE occurrence in this patient. Materials and methods: Laboratory diagnosis of VTE included determination of antiphospholipid antibodies: lupus anticoagulant (LAC) and anticardiolipin antibodies (ACA) IgG and IgM classes, inhibitors of coagulation (antithrombin, protein C, protein S) and fibrinolysis (plasminogen activator inhibitor-1 (PAI-1)), determination of activated protein C resistance (APCR) and molecular diagnosis of thrombophilic factors: genotyping polymorphisms in the gene for PAI-1 and methylenetetrahydrofolate reductase (MTHFR), mutation G20210A for coagulation factor II (prothrombin) and mutation G1691A for coagulation factor V (factor V Leiden). Results: The following antiphospholipid antibodies were demonstrated in the patient: lupus anticoagulant (LAC ratios 3.0 and 2.76) and anticardiolipin antibodies of IgG class (104.7 and 103.7 GPL-U/mL) on two independent measurements nine weeks apart. These results, along with clinical presentation, suggested the presence of antiphospholipid syndrome. Molecular diagnosis confirmed polymorphism in the gene for PAI-1 (genotype 4G/5G), which was in accordance with elevated concentration of PAI-1 measured in plasma (4.9 IU/mL). Conclusion: In this patient, laboratory diagnosis of thrombophilia revealed antiphospholipid syndrome as a risk factor for VTE. Although PAI-1 polymorphism is not considered as an independent risk factor for VTE, it is possible that in patients with established thrombophilic factor such as antiphospholipid syndrome this polymorphism may play a role in VTE occurrence. Infection and prolonged bed rest that preceded thrombosis could have been additional risk factors for VTE occurrence in this patient. The reported case supports the current concepts according to which VTE occurrence most frequently results from interaction of genetic and acquired risk factors.
Biochemia Medica (biochemia-medica@hdmb.hr); Vol.16 No.2.
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ABSTRACT: Background: The aim of this study was to investigate the frequency of C677T methylenetetrahydrofolate reductase (MTHFR) mutation in healthy Croatian volunteers and in patients with atherosclerosis. Methods: The C677T MTHFR gene mutation was determined by polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) in 640 subjects, residents of the Zagreb city or Zagreb surroundings. Control group (n=298) was healthy blood donors. Patients (n=342) were divided into two groups of those with coronary heart disease, CAD (n=247) and those with >60% carotid stenosis, CS (n=95). Results: CC genotype was recorded in 45% of healthy volunteers and 46% of patients (46.3% with CS and 46.2% with CAD). TC genotype was found in 49% of healthy volunteers and 45% of patients (46.3% with CS and 44.9% with CAD). There was no significant difference (p>0.05) from the control group in the genotype or allele frequency either for the overall group of patients with atherosclerosis or for the patient subgroups. Conclusion: The preliminary study of MTHFR polymorphism in control subjects and cardiovascular disease/carotid stenosis patients revealed that in Croats there was a low frequency of TT genotype (6% in controls vs. 9% in patients) and T allele (31% for cases and controls). Additionally, our results did not show significantly higher frequency of MTHFR mutation in CAD and CS studied groups.
Clinica Chimica Acta.
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Biochemia Medica (biochemia-medica@hdmb.hr); Vol.18 No.2.
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ABSTRACT: Severe intrahepatic cholestasis associated with alpha-1-antitrypsin deficiency was confirmed in a newborn. Geno-typing and phenotyping of the newborn and his parents revealed PiZZ genotype in the newborn, PiSZ genotype in his mother and PiMZ in his father. PiSZ genotype is a very rare AAT genotype which was detected in our 7-years of genotyping and 11 years of phenotyping for the first time.
Biochemia Medica (biochemia-medica@hdmb.hr); Vol.17 No.2.
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ABSTRACT: The aim of this study was to determine the association of bleeding as a complication of warfarin therapy with polymorphism of CYP2C9 gene (alleles 1, 2 and 3). The CYP2C9 is the main enzyme for warfarin metabolism. Study included 181 patients receiving warfarin for at least one month. Allele 1 of CYP2C9 gene (in 94.5%) and genotype *1/*1 (57.5%) prevailed. Allele 3 was found in 12.7% patients. Bleeding side-effects occurred in 18 patients (10%). Patients with allele *1 needed significantly higher maintenance warfarin dose (p=0.011). Those with allele *3 had significantly lower maintenance warfarin dose (p=0.005) and higher prothrombin time (PT) at induction (p=0.034). Bleeding occurred significantly more often in those with lower maintenance warfarin dose (p=0.017). Patients with allele *3 had increased risk of bleeding, with marginal significance (p=0.05). Polymorphism of CYP2C9 could determine dose of warfarin therapy and thus it could be related to the risk of bleeding complications. Allele *3 carriers need lower warfarin dose. Therefore, initially reduced warfarin induction dose in allele *3 carriers could avoid more prolonged PT and decrease the risk of bleeding complication.
Collegium Antropologicum (croantro@inantro.hr ); Vol.32 No.2.
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ABSTRACT: Aim: We aimed to evaluate Accu Chek Compact Plus blood glucometer by comparing its accuracy and precision with laboratory reference system. Methods: Original Olympus Glucose reagent on Olympus AU 2700 analyzer served as a reference method. The difference between Olympus and Accu Chek glucose concentrations was tested by paired t-test. Glucometer accuracy was evaluated using: bias (%), Passing-Bablock regression, Bland-Altman and Clarke Error Grid analysis. Intra-assay glucometer precision was examined by 10 consecutive measurements at three glucose levels. Results: The average bias of Accu Chek was -6.6%; Passing-Bablock analysis revealed that there was no significant deviation from linearity; as of Bland-Altman analysis more than 95% of our values lied between mean ± 1.96 SD. The results of the Clarke Error Grid analysis were 100% in zone A of the error grid. Conclusions: Accu Chek Compact Plus blood glucometer has a good accuracy and precision compared with our laboratory referent-sistem and may be used for daily blood glucose self monitoring.
Biochemia Medica (biochemia-medica@hdmb.hr); Vol.18 No.3.
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ABSTRACT: The aim of the study was to investigate the association between methylenetetrahydrofolate (MTHFR) genotypes and levels of homocysteine (Hcy), folate, vitamin B12 and lipids as well as the association between apolipoprotein E (apo E) genotypes and levels of lipids in a Croatian healthy control group and a group of patients with >70% carotid stenosis (CS). The study included 98 Croats, 38 patients with >70% carotid stenosis and 60 age- and sex-matched controls. The MTHFR and apo E genotypes were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), Hcy by enzyme immunoassay, vitamins by immunochemiluminiscence, and lipids by spectrophotometric method. There was no difference between control subjects and CS patients in the distribution of C677T MTHFR genotypes (p=0.786) and alleles (p=0.904), however, differences in the frequencies of apo E genotypes (p=0.012) and alleles (p=0.029) were statistically significant. The odds ratio for apo E 3/4 genotype was 3.93 (95% CI 1.23–12.61). Hyperhomocysteinemia (15 mol/L) was found in 11% of CS patients and 5% of control subjects. Total cholesterol, triglycerides, vitamin B12 and folate were statistically different in »all MTHFR genotypes« (p0.001, p0.01, p=0.044 and p=0.036, respectively), and in TC/TT (p0.001, p=0.003, p=0.030 and p=0.032, respectively) groups. The levels of total cholesterol, LDL cholesterol and triglycerides in the apo E 3/3, and total cholesterol in the apo E 3/4 group yielded statistical difference. An association was found of apo E 3/4 genotype but not of MTHFR genotypes with the risk of CS. MTHFR and apo E affect blood lipid levels, which was statistically confirmed. An association was also recorded between hyperhomocysteinemia and patients with CS. Vitamin status in CS showed a statistically verified association with TC/TT MTHFR genotype. In the group of patients with TC/TT MTHFR genotype, lower vitamin B12 and higher folate values were recorded. The results of multiple logistic analysis showed that there was no statistical significance of Hcy levels (OR 2.403, p=0.334) or conventional vascular risk factors such as smoking habit (OR 0.505, p=0.149), age (OR 1.048, p=0.087) or sex (OR 2.037, p=0.112) in predicting CS.
Collegium Antropologicum (croantro@inantro.hr ); Vol.30 No.4.
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ABSTRACT: Background: Inherited bisalbuminemia is rare, mostly benign state, which has so far been described in several pathological conditions. Two genetic variants have already been described in two Croatian families. Herein we report a new case of bisalbuminemia in a Croatian male patient with sarcoidosis. Methods: The patient was referred by a general practitioner to our hospital for a diagnostic work up for suspected sarcoidosis. Serum capillary protein electrophoresis was performed with automated capillary electrophoresis system. Results: Based on laboratory and pathologic findings, diagnosis of sarcoidosis was confirmed. Serum electrophoresis revealed two distinct albumin bands. Additional albumin variant was slow-migrating. The slow and a normal band made up for 0.48 and 0.51 of the total albumin, respectively. The patient's sister was also bisalbuminemic. Conclusions: This is a new case of inherited bisalbuminemia in a patient with sarcoidosis. Whether bisalbuminemia has any relevance to sarcoidosis, remains to be elucidated. Clinical chemists and clinicians should be alerted by every case of either inherited or acquired albumin variant, because it could provide insight into the protein evolution as well as the physical, chemical and molecular characteristics of albumin.
Biochemia Medica (biochemia-medica@hdmb.hr); Vol.19 No.1.
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ABSTRACT: Introduction: Laboratories accredited according to ISO 15189 standard which have two or more diWerent analyzers or procedures for examinations, should define mechanism for comparison of results. The objective of this study was to present the model for comparison of emergency tests on two different types of automated general biochemistry analyzers. Materials and methods: General biochemistry emergency tests were measured in serum and urine on two automated analyzers Olympus AU2700 and Olympus AU640 (Olympus, Hamburg, Germany) during 60 day period. Olympus AU2700 was used as the reference analyzer and Olympus AU640 as a backup system. Control samples on two levels were used for result comparison. Results: Average 60-day period bias was within acceptance criteria for all analyzes included in this model of comparability assessment. Only low control concentrations for some tests were exceeding allowable biases in less than 2% cases and high level controls were never exceeding allowable biases. Conclusions: This model of results comparison ensures continuous analytical quality of laboratory reports and improves overall efficiency of laboratory services.
Biochemia Medica (biochemia-medica@hdmb.hr); Vol.19 No.3.
