The hero of American and Polish nations: a molecular look at Thaddeus Kosciuszko's cause of death suggests a contribution of endocarditis caused by Cutibacterium acnes infection.

The remains of the heart tissue of Thaddeus Kosciuszko have been investigated as the possible cause of disease and death of the hero of Polish and American nations. Three specimens, DNA isolated from scrappings of wax surface, from the surface of a wooden plate, and from the linen cloth that have had contact with the object were subjected to nanosequencing. From the first two, among all reads identified, only one classified as Propionibacterium acnes (synonymous current name Cutibacterium acnes), had a purported clinical significance. The observed identity between the P. acnes sequences and reference was 89-90% consistent with the hypothesis that the identified reads represent the ancient P. acnes DNA (aDNA), which underwent fragmentation and sequence changes caused by its long-time presence in the environmental conditions conducive to degradation. We present a reasonable and entirely new hypothesis that the analyzed samples could reflect the presence of the bacteria in the original Kosciuszko's heart tissue and that the process of C. acnes infection was progressing inside the organ (endocarditis), not on its surface (pericarditis) leading to rapid deterioration of health and eventually death. We again point out that normal skin and mucosal membranes commensal, a causative agent of common skin acne, may be associated with various severe organ infections posing a threat to health and life.

Identification and In Silico Characterization of a Novel COLGALT2 Gene Variant in a Child with Mucosal Rectal Prolapse.

Rectal prolapse is influenced by many factors including connective tissue dysfunction. Currently, there is no data about genetic contribution in the etiology of this disorder. In this study, we performed trio whole-exome sequencing in an 11-year-old girl with mucosal rectal prolapse and her parents and sibling. Genetic testing revealed a novel heterozygous missense variant c.1406G>T; p.G469V in exon 11 of the COLGALT2 gene encoding the GLT25 D2 enzyme. Sanger sequencing confirmed this variant only in the patient while the mother, father and sister showed a wild-type sequence. The pathogenicity of the novel variant was predicted using 10 different in silico tools that classified it as pathogenic. Further, in silico prediction, according to Phyre2, Project HOPE, I-Mutant3.0 and MutPred2 showed that the missense variant can decrease protein stability and cause alterations in the physical properties of amino acids resulting in structural and functional changes of the GLT25D2 protein. In conclusion, the present study identifies a previously unknown missense mutation in the COLGALT2 gene that encodes the enzyme involved in collagen glycosylation. The clinical features observed in the patient and the results of in silico analysis suggest that the new genetic variant can be pathogenic.

Preliminary analysis of polymorphism of STR markers in the population of domestic cats (Felis catus) in Lower Silesia.

AIM OF THE STUDY: Genetic tests play a crucial role in the crime investigation process and often provide the strongest evidence for case resolution. Although the majority of genetic analyses in the field of criminalistics focus on the human DNA, genetic identification of animals is becoming an increasingly common procedure. Domestic animals, which live around people, may be silent witnesses and even victims of criminal activity. Their typically limited value as evidence in such cases could radically change thanks to the possibility of using animal biological material present at the crime scene. In addition to forensic medicine, genetic identification methods of this type may also become a valuable tool in many other areas of life. Recently, there has been an increase in public interest in verifying the pedigree of animals, investigating poaching and illegal shooting of animals, e.g. protected wildcats and lynx, as well as illegal trade in animals. The main aims of the studies reported in this paper were to assess the degree of polymorphism of the analyzed STR markers in feline genetic material, and to perform a preliminary evaluation of their suitability for developing an original feline genetic identification test. MATERIAL AND METHODS: The studies involved an analysis of genetic material samples obtained from a population consisting of 123 unrelated cats representing various domestic cat breeds, living in the Lower Silesia region. The material collected from individual cats in the form of blood drops or buccal swabs was subjected to an analysis of five STR markers forming a single multiplex assay (FCA742, FCA744, F124, FCA732, FCA749). RESULTS: The results obtained for each marker separately were analyzed statistically and, using the 2 test, the concordance of the study population with the Hardy-Weinberg principle was evaluated. CONCLUSIONS: The findings demonstrate a significant potential of the analyzed markers for the development of genetic identification tests.

First successful DNA isolation and profiling from bone using an approach that is non-destructive toward bone surface

Commonly used destructive DNA isolation techniques consist of pulverization which leads to the complete destruction of smaller bones or irreversible damage to larger bones through the cutting of extensive fragments. The procedure is totally unacceptable when handling bones which are valuable for anthropological or religious reasons, as museum exhibits or due to emotional factors. Since the Department's team has already resolved this problem in the case of human teeth (A. Pilecka), efforts have been undertaken to develop a similar non-destructive technique for the isolation of DNA from human bones. To the best knowledge of the authors of the report, the study has yielded the world's first STR profile derived from DNA isolated from human bones by a technique that is non-destructive toward the bone surface.

PPARs in Clinical Experimental Medicine after 35 Years of Worldwide Scientific Investigations and Medical Experiments.

This year marks the 35th anniversary of Professor Walter Wahli's discovery of the PPARs (Peroxisome Proliferator-Activated Receptors) family of nuclear hormone receptors. To mark the occasion, the editors of the scientific periodical Biomolecules decided to publish a special issue in his honor. This paper summarizes what is known about PPARs and shows how trends have changed and how research on PPARs has evolved. The article also highlights the importance of PPARs and what role they play in various diseases and ailments. The paper is in a mixed form; essentially it is a review article, but it has been enriched with the results of our experiments. The selection of works was subjective, as there are more than 200,000 publications in the PubMed database alone. First, all papers done on an animal model were discarded at the outset. What remained was still far too large to describe directly. Therefore, only papers that were outstanding, groundbreaking, or simply interesting were described and briefly commented on.

Validation of the minisequencing method for detection of G1691A (Leiden) factor V mutation.

INTRODUCTION: The factor V (FV) plays an important role in the coagulation process and belongs to the group of factors that significantly increases the risk of thrombophilia. Our study presents a novel, rapid method for the detection of FV (R506Q) mutation using minisequencing approach. MATERIAL AND METHODS: Samples of peripheral blood were obtained from 300 females of the Lower Silesian population. Minisequencing, as one of the polymerase chain reaction (PCR)-based methods, was used for detection the of FV (R506Q) point mutations. The allele restriction mutation system PCR (ARMS-PCR) verifying method was applied. RESULTS: By using minisequencing reaction we examined the FV genotypes in the female group who experienced at least one unexplained spontaneous miscarriage. The results of the ARMS-PCR, as a verifying test, were fully consistent with the results of the minisequencing technique. DISCUSSION: One of the many factors which may cause thrombophilia is the FV gene mutation R506Q. A full validation of the minisequencing method was carried out in order to apply this method to clinical tests. The validation shows that the minisequencing technique is highly precise and may be used in routine diagnostics of the FV R506Q mutation.

Epigenetic Findings in Twins with Esophageal Atresia.

Esophageal atresia (EA) is the most common malformation of the upper gastrointestinal tract. The estimated incidence of EA is 1 in 3500 births. EA is more frequently observed in boys and in twins. The exact cause of isolated EA remains unknown; a multifactorial etiology, including epigenetic gene expression modifications, is considered. The study included six pairs of twins (three pairs of monozygotic twins and three pairs of dizygotic twins) in which one child was born with EA as an isolated defect, while the other twin was healthy. DNA samples were obtained from the blood and esophageal tissue of the child with EA as well as from the blood of the healthy twin. The reduced representation bisulfite sequencing (RRBS) technique was employed for a whole-genome methylation analysis. The analyses focused on comparing the CpG island methylation profiles between patients with EA and their healthy siblings. Hypermethylation in the promoters of 219 genes and hypomethylation in the promoters of 78 genes were observed. A pathway enrichment analysis revealed the statistically significant differences in methylation profile of 10 hypermethylated genes in the Rho GTPase pathway, previously undescribed in the field of EA (ARHGAP36, ARHGAP4, ARHGAP6, ARHGEF6, ARHGEF9, FGD1, GDI1, MCF2, OCRL, and STARD8).

Assessment of three epigenotypes in colorectal cancer by combined bisulfite restriction analysis.

Recent investigations have demonstrated the clear heterogeneity of sporadic colorectal cancer (CRC) with regard to CpG island methylation. Two unsupervised cluster analyses revealed that CRCs form three distinct DNA methylation subsets, which are referred to as the high-, intermediate-, and low-methylation epigenotypes (HME, IME, and LME, respectively). A recent study by Yagi et al. found a fairly sensitive and specific identification of HME, IME, and LME using two marker panels analyzed by MALDI-TOF mass spectrometry (MassARRAY). However, the expensive equipment required for this method substantially increases the cost and complexity of the assay. In this article, we demonstrate the assessment of HME, IME, and LME in a group of 233 sporadic CRCs using seven markers proposed by Yagi et al. The DNA methylation of each marker was quantified using combined bisulfite restriction analysis (COBRA) and analyzed along with various genetic factors associated with CRC [the BRAF and KRAS mutations, MLH1 methylation and microsatellite instability (MSI)]. The baseline methylation of each marker was generated from pooled DNA isolated from 50 normal colon tissues. We demonstrate that the correlation of HME, IME, and LME epigenotyped by COBRA using different molecular classifiers is similar to that achieved by MassARRAY. Therefore, epigenotyping CRCs using COBRA is a simple, specific, and cost-effective method that has the potential to be widely used in CRC research.

Development of a procedure involving artifact examination to determine the species affiliation of its biological material, as illustrated by an attempt to demystify an alleged Nazi lampshade made from human skin.

Objectives: The main purpose of the study was to identify the species origin of the material from which the incriminating lampshade bought at a flea market had been made. Methods: The histological and molecular biology methods commonly used in forensic genetics were selected to achieve this goal. The DNA for the research was isolated using a QIAamp DNA Mini Kit (Qiagen) according to the manufacturer's protocol for tissues. The quantitative and qualitative evaluation of genetic material was carried out by the real-time PCR method with a Quantifiler Duo DNA Quantification Kit (Applied Biosystems). Specific genetic markers of mtDNA of cattle, equines, deer, wild boar, and sheep were selected to identify species. Results: Histological tests showed that the lampshade had been made from intestinal flaps. The DNA from sample tested positive for cattle. The test results dispelled the suspicion that the researched lampshade had been made from human skin.-hour journey. The second case is a 55-year-old male assaulted with double punches in his chest and declared dead on arrival at the hospital after 30 minutes. A medicolegal autopsy and thorough investigation, in both cases, revealed cardiac tamponade due to ventricular rupture with no underlying pathology. Conclusion: The proposed testing method can be used to verify the origin of the artifacts misleadingly described as made from human skin. To our knowledge, such artifacts can be found in museums and private collections. Further-more, it has been widely believed until now that human-skin products, mainly lampshades, were mass-produced in Nazi concentration camps, mainly in Buchenwald.

[Efficiency of three commercial kits dedicated to DNA and RNA isolation from various clinical and forensic materials using the Janus automated workstation]. / Wydajnosc trzech komercyjnych zestawów do izolacji DNA i RNA ze zróznicowanego materialu klinicznego i dowodowego, przy uzyciu automatycznej stacji Janus.

Isolation of genetic material is a crucial stage in molecular biology. Increasing needs for DNA analysis cause continuous improving of genetic material isolation methods toward higher accuracy and output. Automatization in molecular biology is widely seen, especially in clinical and forensic medicine. The objective of this research was optimization of methods for automatic nucleic acid isolation using the Janus automated workstation, Perkin Elmer. The efficiency and purity of isolated DNA was satisfactory. Despite numerous attempts at achieving automatic RNA isolation, we did not succeed in obtaining RNA working in other applications, such as RT-PCR or Real-Time PCR.

The influence of acute coronary syndrome on levels of clopidogrel active metabolite and platelet inhibition in patients with and without CYP2C19*2(681 G>A), *3(636 G>A) and ABCB1(C3435C> T) gene polymorphisms.

INTRODUCTION: Although ticagrelor and prasugrel remain the standard antiplatelet treatments in acute coronary syndrome (ACS), numerous patients still present with indications for clopidogrel use. AIM: We aimed to assess the levels of clopidogrel active metabolite and to evaluate the effect of the drug on platelet inhibition in patients with ACS as compared with those with stable coronary disease. Patients were assessed for the presence of the most common genetic polymorphisms that reduce the absorption (ABCB1) and activation (CYP2C19*2 and CYP2C19*3) of clopidogrel to exclude the effect of genetic variability on drug concentrations and activity. MATERIAL AND METHODS: This single-center, open-label, prospective study included 199 patients hospitalized due to ST-segment elevation myocardial infarction (STEMI) or non-STEMI (NSTEMI) in Killip class I-III, who underwent percutaneous coronary intervention. The control group included 22 patients with stable coronary artery disease. RESULTS: The mean (SD) levels of active clopidogrel were 17.1 (12.3) ng/ml in controls and 16.4 (12.0) ng/ml in the whole study group (p < 0.68). No differences were noted in clopidogrel levels between patients with STEMI and NSTEMI (mean (SD), 17.6 (2.3) ng/ml and 15.1 (11.5) ng/ml; p < 0.45) or between STEMI and NSTEMI groups and controls (p < 0.38 and p < 0.61, respectively). No effect of ABCB1 or CYP2C19 polymorphism was observed in the study subgroups. CONCLUSIONS: We concluded that ACS does not affect the levels of clopidogrel active metabolite or platelet inhibition in patients in Killip class I-III with or without CYP2C19 or ABCB1 gene polymorphisms.

Original and generic clopidogrel: A comparison of antiplatelet effects and active metabolite concentrations in patients without polymorphisms in the ABCB1 gene and the allele variants CYPC19*2 and *3.

BACKGROUND: Ticagrelor and prasugrel are widely used as antiplatelet therapy after coronary angioplasty. However, there is a group of patients with indications for clopidogrel treatment. This population includes patients with chronic or acute coronary syndrome who are treated invasively and have contraindications to the use of novel antiplatelet drugs due to antithrombotic treatment (particularly with non-vitamin K antagonist oral anticoagulants). A wide range of generic forms of clopidogrel are available on the market. However, it is unclear whether they are as effective as the originator drug. OBJECTIVES: In the current study, we aimed to assess the concentrations of the active metabolite of clopidogrel and its effect on platelet aggregation inhibition in patients receiving the originator drug in comparison with those receiving generic clopidogrel. MATERIAL AND METHODS: We enrolled 22 healthy individuals without polymorphisms in the ABCB1 gene and the allele variants CYPC19*2 and CYPC19*3. All participants received a loading dose of clopidogrel (600 mg), followed by a maintenance dose of 75 mg for the next 3 days. On day 3, blood samples were obtained 1 h after drug administration to assess active metabolite concentrations using liquid chromatography with tandem mass spectrometry. In each participant, platelet aggregation was assessed with light transmission aggregometry after 5-µmol/L and 10-µmol/L adenosine diphosphate (ADP) stimulation. Assays were performed for the originator clopidogrel and 2 different generic groups. RESULTS: The mean ± standard deviation (SD) concentrations of active clopidogrel did not differ between the originator drug and 2 generic products with clopidogrel (12.7±5 pg/µL compared to 13.0 ±4 pg/µL compared to 14.4 ±4 pg/µL). Platelet aggregation inhibition after stimulation with 5 µmol/L and 10 µmol/L ADP was similar for all preparations. CONCLUSIONS: In comparison with original clopidogrel, the use of its generic form does not affect the blood concentrations of the active metabolite or its antiplatelet effect.

Polymorphisms of the MTHFR gene in mothers of children with trisomy 21 (Down syndrome) in a Polish population.

BACKGROUND: Down syndrome (DS) is the most frequent cause of intellectual disability. In 95% of cases, it is caused by simple trisomy of chromosome 21 resulting from nondisjunction of chromosomes in meiotic division. Currently, the molecular and cellular mechanisms responsible for the phenomenon of nondisjunction are unknown. OBJECTIVES: To investigate the incidence of 5 single-nucleotide polymorphisms (SNPs) of the MTHFR gene in a population of Polish mothers who had given birth to children with trisomy 21 in comparison with a control group of women with healthy offspring. MATERIAL AND METHODS: The test material comprised venous blood collected from mothers who had given birth to a child with DS (study group, n = 130) as well as from women who had given birth to children without trisomy 21 (control group, n = 88). DNA was isolated using a kit manufactured by Qiagen. Amplification was carried out using a Qiagen Multiplex PCR Kit (Qiagen); genotyping was performed using SNaPshot Genotyping MasterMix (Applied Biosystems). RESULTS: No statistically significant differences were observed in the frequency of genotypes between the examined groups in terms of the polymorphisms of the MTHFR gene. CONCLUSIONS: In the Polish population studied, no relationship was found between the occurrence of particular genotypes of the MTHFR gene, i.e., 677CT, 1298AC, rs3737964, rs4846048, and rs1994798, in women and the birth of children with trisomy 21. The results contradict the validity of research on polymorphisms of the MTHFR gene as potential predisposing factors for the occurrence of trisomy 21 in children.

Distribution of polymorphisms in the CYP2C19 and ABCB1 genes among patients with acute coronary syndrome in Lower Silesian population.

BACKGROUND: Dual antiplatelet therapy (DAPT) with aspirin and clopidogrel administered to treat patients with acute coronary syndrome (ACS) is still being used. However, despite the proven efficacy of this treatment regimen, thromboembolic complications have been observed in some individuals. The reason for this phenomenon is linked to the so-called increased responsiveness of platelets despite high platelet resistance (HPR). A significant role in HPR is attributed to genetically determined differences in the absorption and activation of clopidogrel. OBJECTIVES: The aim of the study was to assess the incidence of polymorphisms of the ABCB1 and CYPC19 genes that encode proteins involved in the absorption and metabolism of clopidogrel. MATERIAL AND METHODS: The analysis was performed in 199 consecutive patients from Lower Silesian voivodeship (Poland) who underwent coronary angioplasty with stenting for ACS. The single nucleotide polymorphism of the CYP2C19 and ABCB1 genes was performed using a mini sequencing or restriction fragment length polymorphism method. RESULTS: The results of this study revealed the high incidence of patients who may be unresponsive to antiplatelet treatment due to genetic causes. The CYPC19*2 allele in the form of homozygote or mutation heterozygote appeared in 26.1% of the study population. ABCB1 (C3435C> T) polymorphism was associated with 84% of patients. The total incidence of allelic disorders of low drug absorption and metabolism reached 14.6%. CONCLUSIONS: The data obtained should prompt clinicians to use more recent antiplatelet agents (ticagrelor or prasugrel) first, instead of clopidogrel.

Mutation of the MYH7 gene in a child with hypertrophic cardiomyopathy and Wolff-Parkinson-White syndrome.

Familial hypertrophic cardiomyopathy (HCM) displays autosomal dominant inheritance with incomplete penetration of defective genes. Data concerning the familial occurrence of ventricular preexcitation, i.e. Wolff-Parkinson-White (WPW) syndrome, also indicate autosomal dominant inheritance. In the literature, only a gene mutation on chromosome 7q3 has been described in familial HCM coexisting with WPW syndrome to date. The present paper describes the case of a 7-year-old boy with HCM and coexisting WPW syndrome. On his chromosome 14, molecular diagnostics revealed a C 9123 mutation (arginine changed into cysteine in position 453) in exon 14 in a copy of the gene for beta-myosin heavy chain (MYH7). It is the first known case of mutation of the MYH7 gene in a child with both HCM and WPW. Since no linkage between MYH7 mutation and HCM with WPW syndrome has been reported to date, we cannot conclude whether the observed mutation is a common cause for both diseases, or this patient presents an incidental co-occurrence of HCM (caused by MYH7 mutation) and WPW syndrome.

[The Pro12Ala polymorphism of the peroxisome proliferator-activated receptor gamma and immunological processes in patients with type 2 diabetes and insulin resistance]. / Polimorfizm Pro12Ala genu receptora aktywowanego przez proliferatory peroksysomów gamma a procesy immunologiczne w cukrzycy typu 2 wiklanej insulinoopornoscia. Doniesienie wstepne.

INTRODUCTION: The peroxisome proliferator-activated receptor gamma (PPARgamma), a transcriptor factor, regulates immunological and metabolic processes, which are important for carbohydrate and lipid metabolism. Various polymorphic forms of PPARgamma may promote diabetes mellitus and diabetic complications. AIM OF THE WORK: The assessment of TNFalpha gene expression in peripheral blood monocytes, serum TNFalpha concentration and anti-GAD and ICA antibodies in relation to the polymorphism Pro12Ala in patients with 2 diabetes. PATIENTS AND METHODS: 58 patients with type 2 diabetes (average age 59.0 +/- 11 years) and 18 healthy people were examined. The Pro12Ala polymorphism of PPARy gene were assessed using mini-sequence technic SnaPshot [ABIPRISM-310]. The TNFalpha gene expression were estimated using real-time PCR [Applied Bio-systems]. The TNFalpha concentration [Quantikin Immunoassay, R&D Systems] and ICA and GAD antibodies [immunofluorescence method, DRG] were evaluated in venous blood. RESULTS: A heterozygotous genotype Pro12Ala was estimated in 32 patients and a homozygotous genotype Pro12Pro in 21. Only 6 patients were positive for GAD antibodies and only 6 patients for ICA antibodies. The TNFalpha concentration in serum and the TNFalpha gene expression in monocytes did not refer to the Pro12ala polymorphism of PPARy and neither to antibodies. CONCLUSION: 1) The TNFalpha concentration in serum and the TNFalpha gene expression in monocytes do not refer to the Pro12ala polymorphism of PPARgamma in patients with type 2 diabetes. 2) The Pro12Ala genotype do not influence autoimmunologic processes of diabetes.

Serotonin-Related Gene Variants in Patients with Irritable Bowel Syndrome and Depressive or Anxiety Disorders.

AIM: To assess the association of six polymorphisms in serotonin-related genes with depressive or anxiety disorders in patients with irritable bowel syndrome (IBS). METHODS: The lifetime prevalence of depressive and anxiety disorders was assessed in 95 IBS patients (85% women) using the Munich version of the Composite International Diagnostic Interview (CIDI). IBS was diagnosed according to the Rome III criteria. SCL6A4 HTTLPR polymorphism (rs4795541) was determined using PCR-based method. Single-nucleotide polymorphisms in HTR1A (rs6295), HTR2A (rs6313 and rs6311), HTR2C (rs6318), and TPH1 (rs1800532) were detected by minisequencing method. RESULTS: IBS patients with depressive disorders were characterized by higher frequency of 5-HTTLPR L allele in comparison to IBS patients with anxiety disorders. The lower frequency of 1438 A allele in HTR2A was found in IBS patients with depressive disorders in comparison to IBS patients without mental disorders. The lower G allele frequency in HTR2C rs6318 polymorphism among IBS patients with anxiety disorders was also observed. CONCLUSIONS: Our results provide further evidence for the involvement of SLC6A4 rs4795541 and HTR2A rs6311 polymorphisms in the pathophysiology of depressive disorders in IBS patients. The new findings indicate that HTR2C rs6318 polymorphism may be associated with the susceptibility to anxiety disorders in IBS patients.

Single nucleotide polymorphisms in the RET gene and their correlations with Hirschsprung disease phenotype.

Hirschsprung disease (HSCR) is a congenital, heterogeneous disorder, characterized by the absence of intestinal ganglion cells. Recent advances show that the RET gene is a major locus involved in the pathogenesis of HSCR. The aim of this study was to analyse if the HSCR phenotype in the Polish population is associated with the presence of polymorphisms in exons 2, 3, 7, 11, 13, 14 and 15 of the RET gene. Molecular results were compared with clinical and long-term follow-up data in 70 Polish patients with HSCR (84.3% with a short segment and 15.7% with a long segment of aganglionic gut). Single-nucleotide polymorphisms were analysed by using the minisequencing SNaPshot multiplex method. The 135G>A polymorphism in RET exon 2 was overrepresented in HSCR patients, compared with a healthy control group. Moreover, the 135G>A variant was shown to be associated with the severe HSCR phenotype. Two other polymorphisms, 2071G>A in exon 11 and 2712C>G in exon 15, were underrepresented in the patients. The results confirm that these RET polymorphisms play a role in the aetiology of HSCR.

The dependence of serum interleukin-6 level on PPAR-alpha polymorphism in men with coronary atherosclerosis.

BACKGROUND: The association between L162V polymorphism in the gene for peroxisome proliferator activated receptor (PPAR) alpha and the development of coronary heart disease was examined. METHODS: PPAR-alpha polymorphism was determined in 48 men with angiographically confirmed coronary atherosclerosis and in 51 healthy men. RESULTS: The frequency of the V allele of the L162V polymorphism was four times higher in men with atherosclerosis (0.25 in studied group and 0.06 in controls). The polymorphism was not associated with changes in body mass index, lipid pattern, serum adhesion molecules, or vasoactive agents concentrations. The effect of the polymorphism on the serum interleukin-6 level (IL-6) was observed (p<0.01). The serum IL-6 level was higher in homozygotic than in heterozygotic subjects (p<0.02). Multivariate regression analysis showed the existence of a relationship between simvastatin therapy and serum IL-6 level (r=0.83; p<0.05) in the homozygotic men. While in homozygotic patients with atherosclerosis a negative linear correlation between serum IL-6 and NO concentration was shown, in heterozygotic men positive correlations between IL-6 or HDL cholesterol and adhesion molecule levels were found. CONCLUSION: L162V polymorphism in the gene for PPAR-alpha seems to be associated with atherosclerosis through a mechanism including regulation of the IL-6 level.

[Retrospective assessment of the antidepressants tolerance in the group of patients with diagnosis of depression and different CYP2D6 genotype]. / Retrospektywna ocena tolerancji leczenia lekami przeciwdepresyjnymi u pacjentów z rozpoznaniem depresji, z róznym genotypem CYP2D6.

AIM: The majority of antidepressants undergo the oxidative biotransformation catalysed by cytochrome P-450, particularly by izoenzyme CYP2D6, whose activity is genetically determined. In many cases poor tolerance of antidepressants depends on CYP2D6 activity. The aim of the study was the evaluation of the relationship between the CYP2Dg genotype and the occurrence of side effects during antidepressive pharmacotherapy. METHOD: Eighty nine patients were included into study. During the last episode of depression all included patients were treated with antidepressants, whose metabolism is catalysed mainly by CYP2D6. Based on medical records and patient interview the occurrence of side effects was evaluated. The genetic material was isolated from the patients' saliva. Genotyping of CYP2D6 was performed using the PCR techniques. The most frequent inactive alleles in the Caucasian population, *3 and *4 were identified. Alleles that were not identified as *3 or *4 were stated as active allele *1. RESULTS: Based on retrospective analysis among patients treated with antidepressants during the last episode of depression 42 patients (47.2%) reported severe side effects. Comparing to the group of patients with wide type genotype (*1/*1), in the group with the genotype including at least one inactive allele, side effects occurred significantly more frequently. CONCLUSION: In this group, comparing to the group of patients with wide type genotype, severe side effects that required discontinuation of antidepressants also occurred significantly more frequently.