[show abstract][hide abstract] ABSTRACT: To assess lipid profile and the genotype distribution of lipoprotein lipase gene polymorphism at Pvu II polymorphic site within the intron between exons 6 and 7 in patients with hypertriglyceridemia.
Pvu II polymorphism was determined in 116 hypertriglyceridemic patients and 50 normolipidemic controls from Zagreb, Croatia. DNA was extracted from peripheral blood mononuclear cells. Polymerase chain reaction was used for amplification of 6th intron, which was then restricted with Pvu II-restriction endonuclease. Serum lipid and lipoprotein fractions were determined by standard enzymatic methods. Cholesterol concentrations in HDL subfractions, HDL2 and HDL3, were determined after precipitation with polyethyleneglycol. Apolipoproteins (apo) A-I and B were determined by immunonephelometry.
Triglycerides showed a positive correlation with total cholesterol (r=0.222, 95% CI=0.041-0.389, p=0.017) and inverse correlation with HDL-cholesterol (r= -0.278, 95% CI= -0.449 to -0.088, p=0.005), especially with HDL3-cholesterol (r= -0.333, 95% CI= -0.497 to -0.147, p=0.001). The respective frequencies for genotypes /, +/, and +/+ were 22, 58, and 36 in the patient group, and 17, 17, and 16 in the control group. Serum triglycerides in the patient group, expressed as median in mmol/L, were 3.30 (range, 2.60-10.90), 3.60 (range, 2.50-21.50), and 3.99 (range, 2.50-15.56), respectively. Serum concentration of triglycerides differed significantly between the +/+ and / genotype (p=0.043).
There is an association between genetic variation at the locus for lipoprotein lipase and high serum triglyceride levels. This might prove useful in the detection of individuals susceptible to the development of hypertriglyceridemia, as well as a marker in the analysis of this genetic defect in patient families.
Croatian Medical Journal 11/2001; 42(5):517-22. · 1.25 Impact Factor
[show abstract][hide abstract] ABSTRACT: Today, approximately 15% of couples have reduced fertility. In most cases the reason is male infertility, usually of genetic origin. Thus, in the context of research in genes involved in reproduction and sex determination, genetic defects in gametogenesis are being extensively studied. The most frequent pathogenic causes of male infertility are Y chromosomal microdeletions and obstructive azoospermia due to congenital absence of the vas deferens (CAVD) in the presence of mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. We have investigated the most common CFTR gene alterations in Croatian men with CAVD, using Roche research prototype assays. Results revealed that the 5T variant was present in 27% of the subjects. The F508 deletion was found in 21% of the subjects. It was the most frequent mutation, although its incidence was much lower than among patients with cystic fibrosis. The prevalence of microdeletions in the azoospermia factor region (AZF) of the Y chromosome in Croatia was 4.5%. This is the first report of Y microdeletions in the Croatian population. Genetic counseling of all couples with the diagnosis of male infertility is recommended before intrauterine insemination, in vitro fertilization, and intracytoplasmic sperm injection, and should also include AZF and CFTR genotyping. Couples requesting assisted reproductive treatment should be offered molecular analysis of the CFTR gene, if male infertility due to obstructive azoospermia is the underlying cause. Also, men with severe oligozoospermia or non-obstructive azoospermia seeking assisted reproductive treatment should be screened for deletions in the Y chromosome.
Croatian Medical Journal 09/2001; 42(4):416-20. · 1.25 Impact Factor
[show abstract][hide abstract] ABSTRACT: Hearing loss in patients with X-linked agammaglobulinemia is often attributed to recurrent infections. However, recent genetic studies suggest a different etiology in some patients. We present three unrelated patients, 6, 9, and 14 years of age, with large deletions of the terminal portion of the Bruton tyrosine kinase (Btk) gene extending 4.2-19 kb beyond the 3' end of the gene. The DNA immediately downstream of the 3' end of Btk contains the deafness-dystonia protein gene (DDP). Mutations in this gene have recently been shown to underlie the Mohr-Tranebjaerg syndrome, which is characterized by sensorineural deafness, dystonia, and mental deficiency. Besides the immunodeficiency, our patients exhibited progressive sensorineural deafness. The clue to an associated hearing problem was delayed development of speech in one patient and post-lingual deafness noticed between the age of 3-4 years in the other two. These patients have not yet exhibited significant associated neurologic deficits.
Pediatric Allergy and Immunology 05/2001; 12(2):107-11. · 3.38 Impact Factor
[show abstract][hide abstract] ABSTRACT: This study investigated the frequency of angiotensin-converting enzyme (ACE) genotypes, concentrations of total cholesterol (T-C), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), lipoprotein Lp (a), Established Risk Factor (ERF) ratio (total cholesterol/HDL-C), apolipoproteins A-I, A-II, apoBand apoE in 134 menopausal women aged 49.62 +/- 4.83 on oral hormone replacement therapy (HRT) (2 mg 17 beta estradiol plus 1 mg norethisterone acetate/day), during (mean +/- SD) 15.77 +/- 9.94 months. ACE genotypes of 134 menopausal women showed DD genotype in 48 (36%), ID genotype in 59 (44%), and II genotype in 27 (20%) women, with the mean body mass index (BMI) (kg/m2) of 26.34 +/- 4.02, systolic blood pressure (mm Hg) of 145.71 +/- 23.32, diastolic blood pressure of 95.28 +/- 12.88, pulse rate of 77.76 +/- 13.81, positive family history of myocardial infarction (MI) (23%) and stroke (22%); 26% were smokers and 6% consumed alcohol regularly. The mean levels of TC (mmol/l) were 5.72 +/- 1.25, TG (mmol/L) 1.63 +/- 0.82, HDL-C (mmol/L) 1.15 +/- 0.29, LDL-C (mmol/L) 3.98 +/- 1.31, lipoprotein Lp(a) (g/L) 0.16 +/- 0.24, ERF ratio 5.35 +/- 1.90, apolipoproteins (g/L): A-I 1.83 +/- 0.39, A-II 0.57 +/- 0.12, apoB 0.92 +/- 0.31, and apoE 0.08 +/- 0.04. The highest mean levels of T-C 5.89 +/- 1.40, TG 1.67 +/- 0.96, LDL-C 4.15 +/- 1.60, lipoprotein Lp(a) 0.19 +/- 0.25) apoB 0.95 +/- 0.32 and ERF ratio 5.46 +/- 2.24 were found in ID genotype, while in DD genotype HDL-C 1.11 +/- 0.28 and apo A-I 1.78 +/- 0.34 were lowest. In II genotype, the levels of apo A-II 0.56 +/- 0.11 were lowest and of apoE 0.09 +/- 0.05 highest. According to DD, ID and II genotypes and lipid, lipoprotein Lp(a), ERF ratio and apolipoprotein concentrations, there were no statistically significant differences between groups. ERF ratio in DD genotype showed a positive correlation with TG (r = 0.59) and LDL-C (r = 0.57), a slight positive correlation with apoB (r = 0.40), and a strong negative correlation with HDL-C (r = -0.73). ERF in ID genotype showed a strong negative correlation with HDL-C (r = -0.73), strong positive correlation with TG (r = 0.70), and T-C (r = 0.58), and slight positive correlation with LDL-C (r = 0.36) and alcohol abuse (r = 0.34). In II genotype, ERF ratio showed a strong positive correlation with LDL-C (r = 0.73), T-C (r = 0.70) and apoE (r = 0.58), slight positive correlation with apoB (r = 0.46) and TG (r = 0.36), and negative correlation with HDL-C (r = -0.54). Matrix correlation of DD genotypes showed the highest positive correlation between T-C and LDL-C (r = 0.91) and apoE (r = 0.45), and negative correlation between HDL-C and ERF ratio (r = 77), and LDL-C and ERF ratio (r = 0.55). In ID genotype, T-C showed a strong positive correlation between LDL-C (r = 0.75) and ERF ratio (r = 0.63), TG and ERF ratio (r = 0.73), and negative with HDL-C (r = 0.53). In genotype II, T-C showed a strong positive correlation between LDL-C (r = 0.96), ERF ratio (r = 0.71), apoB (r = 0.66) and apoE (r = 0.46). LDL-C correlated positively with ERF ratio (r = 0.72), apoB (r = 0.61) and apoE (r = 0.48). These findings indicated the frequency of ACE genotypes to differ within the group of menopausal women. Analysis of ACE genotypes showed ID genotype to be most common among menopausal women. This result indicated their intermediate risk of coronary heart disease (CHD) and myocardial infarction (MI). It has been well established that an increased risk of MI is associated with high frequency of DD genotype, and a low risk with high frequencies of II genotype. In addition to ACE polymorphism analysis, assessment of lipid, apolipoprotein, and lipoprotein Lp(a) concentrations, and of ERF ratio provides further important parameters for better understanding of the risk factors for CDH in women. In the present study, assessment of the genetic, metabolic and environmental markers pointed to an intermediate risk of CHD in menopausal women on HRT, although the mechanism underlying the disease is not clear and well understood yet.
Acta medica Croatica: c̆asopis Hravatske akademije medicinskih znanosti 02/2001; 55(4-5):161-7.
[show abstract][hide abstract] ABSTRACT: The renin-angiotensin-aldosterone system (RAAS) has been considered one of the probable pathophysiologic mechanisms involved in disease progression. Genetic polymorphism of the RAAS has been associated with the clinical course of renal disease. One of the genetic polymorphisms is a deletion or insertion of a 287 base pair fragment in intron 16 of the angiotensin-converting enzyme (ACE) gene. It is known that ACE gene polymorphism is present in humans and that it is associated with an increased risk of cardiovascular diseases, renal disease progression and sarcoidosis. In this study, the potential significance of ACE gene polymorphism in patients with systemic lupus erythematosus (SLE) was investigated. ACE gene polymorphism was determined in 18 patients with SLE and in 21 healthy volunteers as a control group. The mean age of patients was 38.5 years. All patients had a mean follow-up of 30.7 +/- 20.2 months (range 5-95 months). ACE genotypes were determined by the method of polymerase chain reaction. Proteinuria and creatinine were also followed. The frequency of DD, ID and II genotypes was 50%, 28% and 22% in SLE patients and 25%, 50% and 25% in healthy controls, respectively. DD genotype was more common in SLE patients than in the control group. The patients with II genotype had lower proteinuria and creatinine level than those with DD genotype (p < 0.05). The time to disease remission was shorter in patients with II genotype (p < 0.05). Study results indicated an increased frequency of D allele in SLE patients. The increased ACE activity in these patients pointed to the need of further studies of ACE gene polymorphism in SLE.
Acta medica Croatica: c̆asopis Hravatske akademije medicinskih znanosti 01/2001; 55(2):73-6.
[show abstract][hide abstract] ABSTRACT: We report a large genomic deletion of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, viz., a deletion that is frequently observed in Central and Eastern Europe. The mutation, termed CFTRdele2,3(21 kb), deletes 21,080 bp spanning introns 1-3 of the CFTR gene. Transcript analyses have revealed that this deletion results in the loss of exons 2 and 3 in epithelial CFTR mRNA, thereby producing a premature termination signal within exon 4. In order to develop a simple polymerase chain reaction assay for this allele, we defined the end-points of the deletion at the DNA sequence level. We next screened for this mutation in a representative set of European and European-derived populations. Some 197 CF patients, including seven homozygotes, bearing this mutation have been identified during the course of our study. Clinical evaluation of CFTRdele2,3(21 kb) homozygotes and a comparison of compound heterozygotes for (F508/CFTRdele2,3(21 kb) with pairwise-matched (F508 homozygotes indicate that this deletion represents a severe mutation associated with pancreatic insufficiency and early age at diagnosis. Current data show that the mutation is particularly common in Czech (6.4% of all CF chromosomes), Russian (5.2%), Belorussian (3.3%), Austrian (2.6%), German (1.5%), Polish (1.5%), Slovenian (1.5%), Ukrainian (1.2%), and Slovak patients (1.1%). It has also been found in Lithuania, Latvia, Macedonia and Greece and has sporadically been observed in Canada, USA, France, Spain, Turkey, and UK, but not in CF patients from Bulgaria, Croatia, Romania or Serbia. Haplotype analysis has identified the same extragenic CF-haplotype XV-2c/KM.19 "A" and the same infrequent intragenic microsatellite haplotype 16-33-13 (IVS8CA-IVS17bTA-IVS17bCA) in all examined CFTRdele2,3(21 kb) chromosomes, suggesting a common origin for this deletion. We conclude that the 21-kb deletion is a frequent and severe CF mutation in populations of Eastern- and Western-Slavic descent.
Human Genetics 02/2000; 106(3):259-268. · 4.63 Impact Factor
[show abstract][hide abstract] ABSTRACT: Objective: We analyzed correlation between ACE gene polymorphism and adaptive hypertrophic response of LV with regard to BP in competitive oarsmen.Design and methods: ACE gene polymorphism was determined using PCR method in 52 oarsmen with mean age 26 years (range 18–37) and in age, sex and ethnically matched control group of 34 persons. The LV mass was determined echocardiographically and calculated after Devereux. BP was measured using both mercury sphingomanometer and Ambulatory BP Monitor SpaceLabs 90207.Results: The distribution of ACE gene polymorphism was: in competitive oarsmen 16 (47.1%) II, 12 (35.3%) ID, 12 DD (17.6%); in control group 8 (24%) II, 18 (52%) ID and 8 (24%) DD. The frequency of I allele was significantly greater in oarsmen (0.65 vs. 0.50; p 0.05), while in oarsmen with DD genotype systolic BP in nighttime was significantly greater compared to persons with II genotype (121.8 ± 7.4 vs. 114.0 ± 7.0; p 0.05). No changes in geometrical structure of LV regarding ACE genotype were established. However, remodeling of LV was associated with increased variability of systolic and diastolic BP.Conclusion: Observed greater frequency of I allele in competitive rowers could suggest that ACE gene polymorphism might contribute to the athletic performance. According to our results DD genotype and BP variability influences LV hypertrophic response in competitive athletes.
[show abstract][hide abstract] ABSTRACT: Objective: To test the hypothesis that polymorphism of angiotensin converting enzyme (ACE) gene I/D polymorphism is linked to the atheromatous renal artery stenosis.Design and methods: ACE gene polymorphism was determined using PCR method in 40 Caucasian patients with angiographically confirmed renal artery stenosis compared to 73 control subjects matched for age and gender from the same ethnic group. Presence of other risk factors (cholesterol, tryglicerides, serum glucose, smoking) was also analyzed as well as the severity of hypertension. History of cardiovascular or cerebrovascular disorders was also analyzed.Results: The distribution of ACE gene polymorphism was II 13 (32.5%), ID (40%) 16 DD 11 (27.5%); and controls II 18 (24.8%), ID 31 (42.4%), DD 24 (32.8%) (p > 0.05). Therefore, the frequency of D allele was not significantly different from the control group (p > 0.05). We failed to find significant difference in blood pressure values between particular genotypes (p > 0.05). There were also no differences in fasting serum glucose, cholesterol and trigliceryde levels between patients with DD genotype and others (p > 0.05). Also no association was found between ACE gene polymorphism and ECG findings and fundus examination (p > 0.05). However, although not statistically significant (p > 0.05), duration of hypertension was the shortest in patients with DD genotype (7.7 ± 3.8 vs. 11.3 ± 2.7 in II and vs. 9.6 ± 1.8 in ID).Conclusion: According to our results ACE gene polymorphism does not seem to have a predisposing role in the development of atheromatous renal artery stenosis. However, the observed shorter duration of hypertension in patients with DD genotype could implicate that it might have some influence in the progression of this disease.
[show abstract][hide abstract] ABSTRACT: Presentation of a prenatally diagnosed case of Werdnig-Hoffmann disease, the most severe type of spinal muscular atrophy.
DNA obtained from cultivated amniocytes was analyzed for deletions in the survival motor neuron gene and neuronal apoptosis inhibitory protein gene.
The fetus was diagnosed as an affected homozygote for deletions in exon 7 and exon 8 of the survival motor neuron gene. No deletions of exon 5 in the neuronal apoptosis inhibitory protein gene were found.
Direct DNA deletion analysis of the survival motor neuron gene and neuronal apoptosis inhibitory protein gene in affected families represents a highly reliable and fast method for prenatal diagnosis of Werdnig-Hoffmann disease.
Croatian Medical Journal 10/1999; 40(3):433-7. · 1.25 Impact Factor
[show abstract][hide abstract] ABSTRACT: Fetal echoic bowel can be a normal second trimester ultrasonographic finding which usually disappears by 20 weeks on serial sonograms. Recent studies have suggested a possible association of hyperechoic fetal bowel with chromosomopathies and cystic fibrosis. The aim of our study is to determine the incidence of chromosomopathies and cystic fibrosis mutations among the fetuses with isolated hyperechoic bowel.
Sixteen fetuses with isolated echoic bowel were detected: 13 fetuses < or =20 weeks gestation (group I) and 3 fetuses at 20-26 weeks gestation (group II). Cytogenetic studies were performed in all 16 cases and 11 families had deoxyribonucleic acid-based risk assessment for cystic fibrosis. The echogenity of bowel was that of surrounding bone.
Two cases of trisomy 21 and 1 case of trisomy 13 were detected (18.7%). The other ultrasonographic markers begin to appear after 21 weeks gestation in fetuses with trisomy 13. Two of 3 pregnant women with pathological karyotype were younger than 35 years. One of 11 cases (9%) was found to be a heterozygote carrier for deltaF508 mutation.
Isolated hyperechoic bowel in the second trimester was found to be associated with a significantly higher risk of fetal aneuploidy.
The Journal of Maternal-Fetal Medicine 01/1999; 8(2):44-7.
[show abstract][hide abstract] ABSTRACT: Childhood onset proximal spinal muscular atrophy presents with considerable clinical variability. This study included 14 Croatian children aged 11 days to 8 years with spinal muscular atrophy types I-III verified clinically and electromyoneurographically. DNA of affected children was screened for deletions of exons 7 and 8 of the survival motor neuron gene and for deletion of exon 5 of the neuronal apoptosis inhibitor protein gene. Motor nerve conduction velocity and compound muscle action potential amplitude were decreased in children with spinal muscular atrophy type I and II. Deletions of exons 7 and 8 of the survival motor neuron gene and of exon 5 of the neuronal apoptosis inhibitor protein gene in children with spinal muscular atrophy type I-II suggested existence of more genetic abnormalities as compared to type III. A decrease in compound muscle action potential amplitude and motor nerve conduction velocity in children with spinal muscular atrophy correlated with the disease severity, probably as a result of axonal degeneration. Phenotypic severity in children onset spinal muscular atrophy is directly correlated with the extent of survival motor neuron and neuronal apoptosis inhibitor protein exon deletions.
Clinical Chemistry and Laboratory Medicine 09/1998; 36(8):667-9. · 3.01 Impact Factor
[show abstract][hide abstract] ABSTRACT: Apo E genotypes and plasma metabolic risk factors (total cholesterol, triglycerides, HDL and LDL cholesterol, total/HDL cholesterol ratio, lipoprotein Lp (a), apolipoprotein A-I, A-II, apo B, and apo E) were determined in 134 healthy middle-aged (X +/- SD 49.62 +/- 4.83) women. The aim of this study was to investigate metabolic risk markers according to various apo E genotypes, and to evaluate a possible risk for coronary heart disease. The results revealed that the frequencies of apo E3/3 are the most frequent (46%), followed by E4/4 (2%), E3/4 (14%), E2/3 (14%), and E2/4 (2%) in the middle-aged women. Higher mean triglycerides, LDL-C and apo B levels were found with apo E3/4, and lower mean levels of HDL-C i.e. apo A-I than in other analyzed genotypes. Greater mean of total/HDL ratio and lower levels of apo A-II were seen with E2/4. Serum lipoprotein Lp (a) concentration was higher in women with genotypes E3/3. Apo E concentration was the lowest with genotypes E4/4, i.e. the highest with E2/3. Serum total cholesterol tended to be higher in women with genotypes E4/4. Genotype E3/4 is connected with the highest concentrations of (X +/- SD) triglycerides (1.74 +/- 0.78), LDL (4.28 +/- 1.88), apo B (1.03 +/- 0.32) and with the lowest concentrations of HDL cholesterol (1.11 +/- 0.21) in the relation to the other analyzed genotypes. This group of women could possibly represent high risk women for CHD. Genotype E3/3 is associated with the highest concentration of independent genetic risk marker for CHD, lipoprotein Lp (a) (0.19 +/- 0.27). The genotype E4/4 has the highest concentration of total cholesterol (5.93 +/- 1.01), and has to be taken in account for risk evaluation in women. High level of apo E (0.11 +/- 0.05) and low level of apo A-I (1.80 +/- 0.44) were associated with E2/3 genotypes. The significance of E3/4 with the high total/HDL ratio (5.52 +/- 2.21) and low apo A-II (0.53 +/- 0.09) is important indicator, because total/HDL cholesterol ratio represents independent Established Risk Factor (ERF) for CHD. Apolipoprotein E genotypes as genetic markers and investigation of serum metabolic risk markers appear to be important in view for further evaluation of high risk women for CHD in our population.
Collegium antropologicum 07/1998; 22(1):149-55. · 0.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: Two genes, i.e. survival motor neuron (SMN) and neuronal apoptosis inhibitory protein (NAIP) have been mapped to the SMA region of chromosome 5q13. Both genes are frequently deleted or truncated in SMA patients. We have studied 26 patients with SMA types I-III, 29 first relatives, and 14 subjects with mild adult-onset type IV. DNA deletion genotypes were determined by PCR techniques amplifying exons 7 and 8 of SMN, and exon 5 of NAIP gene which distinguish SMN and NAIP telomeric copy from a non-pathogenic gene homologue as a centromeric copy. Results revealed the homozygous deletions of exon 7 and 8 of the SMN gene and exon 5 of the NAIP gene in 3/3 infants with SMA I and in 1/20 with SMA type II. Exons 7 and 8 of the SMN gene were homozygously deleted in 10/20 and only exon 7 in 6/20 children with SMA type II. The overall percentage of deletion cases observed was 77% in children with SMA types I-III. Adult patients with type IV SMA showed no homozygous deletion of exons 7, 8 and 5 of the SMN and NAIP genes. Also, all relatives had both a telomeric and centromeric SMN and NAIP copy. Deletion analysis of SMN and NAIP genes are a significant diagnostic tool, because there are clinical entities resembling SMA which most likely have another pathogenetic background.
Collegium antropologicum 01/1998; 21(2):487-92. · 0.61 Impact Factor
[show abstract][hide abstract] ABSTRACT: The dystrophin gene deletion in 53 Duchenne and 21 Becker muscular dystrophy (DMD/BMD) male patients was analyzed by DNA test using multiplex polymerize chain reaction (M-PCR) in Croatian population. The overall percentage of deletion cases observed was 50%; 61% (53/32) for DMD and 38% (21/8) for BMD. The number of deleted exons was variable, but generally DMD deletions involving single-exon 19, 44, 50, 51 and larger exon deletions 3-6, 4-12, 4-17, 8-13, 12-13, 12-19, 48-50, 50-51, 50-52, 51-52 were more frequent. Eight patients with BMD had deletions exon 45-47, 45-48, and exon 3. The results obtained in the present study showed location of breakpoints in the dystrophin gene, and pointed to variability of deletion patterns in Croatian population among different European populations.
Collegium antropologicum 07/1997; 21(1):151-6. · 0.61 Impact Factor