[Show abstract][Hide abstract] ABSTRACT: Introduction:
Standard pre-operative diagnosis of nodular goitre is not always conclusive. The decision about nodular goitre surgery is increasingly based on molecular methods. The aim of the study was to determine BRAF T1799A mutation and KRas proto-oncogene mutation, and the analysis of RASSF1A promoter methylation level in cytological material obtained from FNAB specimens of thyroid nodules.
Material and methods:
The study population consisted of 85 women and 12 men. The study material was genomic DNA isolated from peripheral blood and thyroid bioptates. Pyrosequencing was used for the evaluation of RASSF1 methylation level. KRas mutation was investigated with Sanger sequencing. BRAF mutation was analysed by standard methods of real-time amplification detection (real-time PCR) with the use of specific starters surrounding the mutated site.
A significant positive correlation was demonstrated between mean methylation of four CpG islands of RASSF1A gene and thyroid tumour volume and its largest diameter (p < 0.05). KRas mutation was not detected in any of the 97 patients. In 7/85 subjects (8.2%) BRAF mutation was observed. In 6/7 patients with BRAF mutation, FNAB of thyroid nodules confirmed a benign nature of the lesions; the material was non-diagnostic in one patient, and papillary thyroid cancer was diagnosed on the basis of postoperative histopathology assessment.
The results of genetic tests reported in our study indicate that the presence of BRAF mutation or higher RASSF1A methylation levels in FNAB cytology specimens of benign lesions may be useful in the assessment of oncological risk, while the evaluation of KRas proto-oncogene mutation is not a valuable test in pre-operative diagnosis of nodular goitre. (Endokrynol Pol 2015; 66 (5): 384-393).
Preview · Article · Oct 2015 · Endokrynologia Polska
[Show abstract][Hide abstract] ABSTRACT: The outcome of kidney allograft transplantation is associated with numerous donor-dependent and recipient-dependent immunological and non-immunological factors. Studies on genetic factors affecting the non-immunological aspects, like ageing of the kidney allograft and patient outcome are still lacking. The aim of this study was the analysis of relative telomere length (RTL; T/S ratio) in the biopsy specimens of the transplanted kidney allograft and its correlation with the delayed graft function (DGF), acute rejection (AR) and chronic allograft dysfunction (CAD).
The study enrolled 119 Caucasian kidney allograft recipients (64 M/55 F, mean age 47.32 ± 14.03; transplantation performed between 2001 and 2012). Organs were harvested from cadaveric donors (59.8 M/40.2 F, mean age 45.99 ± 14.62).
There were significant differences in RTL assessed in kidney allograft biopsy specimens collected 3-6 months after transplantation between patients with DGF and without DGF (181.8 ± 82.0 vs. 284.6 ± 149.6; p < 0.05) and in RTL of kidney allograft biopsy specimens collected 18-60 months after transplantation between patients with AR and without AR (188.1 ± 162.1 vs. 263.3 ± 134.7; p = 0.047). There were significant differences in RTL assessed in kidney allograft biopsy specimens collected 12-24 months after transplantation between patients with CAD and without CAD (168.0 ± 120.0 vs. 282.1 ± 158.4; p = 0.038).
Duration of dialysis before transplantation and PRA influence the kidney allograft ageing. Telomere length assessed in biopsy specimens collected in the peri-transplant period predicts the long-term kidney allograft function. Complications of kidney transplantation, like DGF, AR and CAD are linked with the telomere length and thus, graft ageing.
[Show abstract][Hide abstract] ABSTRACT: To assess the association between PTPN22 1858C>T gene polymorphism and susceptibility to, and clinical presentation of, systemic lupus erythematosus (SLE). Our study included 135 SLE patients (120 women and 15 men; mean age 45.1 years; mean course of disease from 0.5 to 31 years) and 201 healthy subjects. The PTPN22 1858C>T gene polymorphism was genotyped by polymerase chain reaction restriction fragment length polymorphism. A significantly higher incidence of genotype CT in patients with SLE (36.3 %) was found, compared with the control group (24.9 %). The frequencies of C1858 and T1858 alleles were 78.1 and 21.9 % in SLE patients and 86.1 and 13.9 % in controls, respectively. Significantly higher SLE susceptibility was observed in patients carrying at least one T allele (p = 0.009; OR 1.86; 95 % CI 0.14-3.05). Significant association of the PTPN22 T1858 allele (CT + TT vs.CC) and secondary antiphospholipid syndrome was observed (p = 0.049). In SLE patients carrying the T1858 allele, higher levels of antiphospholipid antibodies (anticardiolipin antibodies and/or lupus anticoagulant) were found (p = 0.030; OR 2.17; 95 % CI 1.07-4.44).
[Show abstract][Hide abstract] ABSTRACT: Preoperative diagnostic investigations of nodular goiter are based on two main examinations: ultrasonography of the thyroid gland and ultrasound-guided fine-needle aspiration biopsy. So far, FNAB has been the best method for the differentiation of nodules, but in some cases it fails to produce a conclusive diagnosis. Some of the biopsies do not provide enough material to establish the diagnosis, in some other biopsies cytological picture is inconclusive.
Determining the eligibility of thyroid focal lesions for surgery has been more and more often done with molecular methods. The most common genetic changes leading to the development of thyroid cancer include mutations, translocations and amplifications of genes, disturbances in gene methylation and dysregulation of microRNA. The mutations of Ras proto-oncogenes and BRAF gene as well as disturbances of DNA methylation in promoter regions of genes regulating cell cycle (e.g. hypermethylation of RASSF1A gene and TIMP-3 gene) play an important role in the process of neoplastic transformation of thyreocyte. The advances in molecular biology made it possible to investigate these genetic disturbances in DNA and/or RNA from peripheral blood, postoperative thyroid tissue material and cytology specimens obtained through fine-needle aspiration biopsy of focal lesions in the thyroid gland.
As it became possible to analyze the mutations and methylation of genes from cell material obtained through fine-needle aspiration biopsy, it would be beneficial to introduce the techniques of molecular biology in the pre-operative diagnosis of nodular goiter as a valuable method, complementary to ultrasonography and FNAB. The knowledge obtained from molecular studies might help to determine the frequency of follow-up investigations in patients with nodular goiter and to select patients potentially at risk of developing thyroid cancer, which would facilitate their qualification for earlier strumectomy.
[Show abstract][Hide abstract] ABSTRACT: The aim of our study was to examine NPHS1, NPHS2, WT1 and LAMB2 mutations, previously reported in two thirds of patients with nephrotic syndrome with onset before the age of one year old. Genomic DNA samples from Polish children (n=33) with Steroid-Resistant Nephrotic Syndrome (SRNS) due to focal segmental glomerulosclerosis (FSGS), manifesting before the age of 13 years old, underwent retrospective analysis of NPHS1, NPHS2, WT1 (exons 8, 9 and adjacent exon/intron boundaries) and LAMB2. No pathogenic NPHS1 or LAMB2 mutations were found in our FSGS cohort. SRNS-causing mutations of NPHS2 and WT1 were detected in 7 of 33 patients (21%), including those with nephrotic syndrome manifesting before one year old: five of seven patients. Four patients had homozygous c.413G>A (p.Arg138Gln) NPHS2 mutations; one subject was homozygous for c.868G>A (p.Val290Met) NPHS2. A phenotypic female had C>T transition at position +4 of the WT1 intron 9 (c.1432+4C>T) splice-donor site, and another phenotypic female was heterozygous for G>A transition at position +5 (c.1432+5G>A). Genotyping revealed a female genotypic gender (46, XX) for the first subject and male (46, XY) for the latter. In addition, one patient was heterozygous for c.104dup (p.Arg36Profs*34) NPHS2; two patients carried a c.686G>A (p.Arg229Gln) NPHS2 non-neutral variant. Results indicate possible clustering of causative NPHS2 mutations in FSGS-proven SRNS with onset before age one year old, and provide additional evidence that patients with childhood steroid-resistant nephrotic syndrome due to focal segmental glomerulosclerosis should first undergo analysis of NPHS2 coding sequence and WT1 exons 8 and 9 and surrounding exon/intron boundary sequences, followed by gender genotyping.
No preview · Article · May 2014 · Bosnian journal of basic medical sciences / Udruzenje basicnih mediciniskih znanosti = Association of Basic Medical Sciences
[Show abstract][Hide abstract] ABSTRACT: The role of adenosine as a cardioprotective agent is well known and recent experimental studies suggest that impairment of adenosine-related signal transduction contributes to the pathophysiology of chronic heart failure. The recent observation of an association between ADA, genetic polymorphism and coronary artery disease (CAD) prompted us to study the possible relevance of three intragenic polymorphic sites of the ADA gene (ADA1, ADA2 and ADA6).
136 non-diabetic patients with coronary artery disease and 246 healthy blood donors from the white Italian population of Central Italy and 129 non-diabetic patients with CAD and 204 newborns from the white Polish population were studied. ADA1, ADA2 and ADA6 genotypes were determined by DNA analysis. In males, the proportion of ADA1 *2 (P = 0.0001) and ADA2 *2 (P = 0.005) alleles is lower in CAD than in controls. In males, the haplotype distribution of the pairs ADA1-ADA2, ADA1-ADA6 and ADA2-ADA6 shows statistically significant differences between coronary artery disease and controls.
The present study suggests a complex association between ADA gene and coronary artery diseases. Besides the control of adenosine concentration due to deamination of adenosine, also other functions of the ADA gene could have a role in the susceptibility and/ or clinical course of coronary artery disease.
No preview · Article · Feb 2014 · Acta cardiologica
[Show abstract][Hide abstract] ABSTRACT: Background
This study analyzes the influence the of kidney donor hemostasis on the risk of complications in the kidney recipient after transplantation.
We enrolled 38 deceased kidney donors, of whom14 donors died from a physical injury and the others died from ischemic or bleeding central nervous system stroke. The donors were categorized into 2 subgroups. If the recipient’s postoperative period proceeded smoothly, the kidney donor was assigned to the uncomplicated donors (UD) group. If the recipient’s postoperative period was complicated, the donor was assigned to the complicated (CD) Group. The CD group of consisted of 9 donors who died from strokes or bleedings and 2 who died from physical injury. We examined the antithrombin (AT) protein C (PC), complexes of thrombin/antithrombin (TAT), fragments F1+2 of prothrombin (F1+2), plasminogen (Pl), complexes of plasmin/antiplasmin (PAP), and D-dimers (D-d).
In the CD group had decreased activity of AT, PC, and Pl and increased activity of F1+2, TAT, and D-d. The UD group had a higher level of PAP. The CD group had evidence of intensive blood coagulation, but the UD group had evidence of fibrinolysis. Fisher’s exact test revealed an increased risk in recipients who received a kidney from the CD group.
The hemostasis of the kidney donors had a correlation with the occurrence of some complications in the kidney recipients, especially complications connected with activation of blood coagulation. It seems that the activation of fibrinolysis could be positive prognostic factor, but this requires further investigations.
Full-text · Article · Dec 2013 · Medical science monitor: international medical journal of experimental and clinical research
[Show abstract][Hide abstract] ABSTRACT: Calmodulin II (CALM2) gene polymorphism might be responsible for the variation in the left ventricular mass amongst healthy individuals. The aim was to evaluate the correlation between left ventricular mass (LVM) and g.474955027G>A (rs7565161) polymorphism adjacent to the CALM2 gene.
Healthy Polish newborns (n = 206) were recruited. Two-dimensional M-mode echocardiography was used to assess LVM. Polymorphisms were determined by polymerase chain reaction-restriction fragment length polymorphism and sequencing analyses.
The carriers of the G allele of the CALM2 polymorphism had significantly higher left ventricular mass/weight (LVM/BW) values, when compared with newborns homozygous for the A allele (3.1 g/m2 versus 2.5 g/m2, Padjusted = 0.036). The AG genotype of CALM2 was associated with the highest values of LVM/BW, exhibiting a pattern of overdominance (2.9 g/kg versus 3.1 g/kg versus 2.5 g/kg, Padjusted = 0.037).
The results of this study suggest that G>A CALM2 polymorphism may account for subtle variation in LVM at birth.
Full-text · Article · Nov 2013 · International Journal of Genomics
[Show abstract][Hide abstract] ABSTRACT: In this publication would like to present the Polish Genetic Base of Totalitarianism Victims. On 28th September 2012, the Pomeranian Medical University in Szczecin and the Institute of National Remembrance-Commission for Prosecution of Crimes Against the Polish Nation signed an agreement on the Polish Genetic Base of Totalitarianism Victims (PBGOT, www.pbgot.pl). Our aim is to use the latest methods of forensic genetics to identify the victims of totalitarian systems. The project assumes obtaining proper reference material from the next-of-kin of victims of communist and Nazi crimes. Due to the time passage this has become the most important task for PBGOT. Work includes also securing and storing biological material collected during the exhumation of the victims, and carrying out the genetic identification process. The next task is collecting all available historical data concerning killed and missing people. All these procedures had been already applied in practice during the first stage of identifying victims of communists buried on Powązki Military Cemetery in Warsaw. At this cemetery hundreds of people were buried after they had been killed through military court orders during the years 1948-1956. This number includes most of the members of the Polish resistance movement. In 2012 and 2013 exhumations were carried out and resulted in finding remains of 194 people. A team of specialists in the fields of history, archeology, forensic anthropology, forensic medicine and genetics, working on several scientific units from all over Europe, takes part in this research. The main goal is to identify the victims and so far, remains of 16 exhumed people were identified.
[Show abstract][Hide abstract] ABSTRACT: Limited studies indicate a possible association of 5'-UTR thymidylate synthase enhancer region polymorphism and treatment outcome in patients medicated with 5-fluorouracil (5-FU). The study was designed to verify the relationship in patients with colorectal cancer (CRC), a Polish population that received 5-FU-based adjuvant chemotherapy. The study analyzed 145 Astler-Coller B2 and C CRC patients. Genotyping for a variable number of tandem repeats and G to C single-nucleotide polymorphism in the 5'-UTR of the thymidylate synthase (TS) gene was carried out. TS genotypes were classified into high expression (high TS) and low expression types (low TS). High TS was found in 22.8% of patients. The right-side tumors were more frequently associated with high TS than the left-side tumors (p=0.024). High TS was only found in 9.3% of rectal tumors, but in 29.7% of colon cancers (p=0.0042). Disease-free survival after 20 months (DFS 20) was longer in subjects with low TS than in high TS (p=0.043). Patients who underwent chemotherapy had longer DFS 20 in the low TS than in the high TS subgroup (p=0.051). The low TS was found to be an independent good prognostic factor for DFS 20 in the whole group as well as in the subgroup treated with chemotherapy (p=0.024 and p=0.034, respectively). Patients with low TS did not show any differences in DFS 20 whether they were treated with adjuvant chemotherapy or not. Proximal CRC tumors are characterized by higher TS expression genotypes than distal tumors, and are at significantly greater risk of early recurrence during the first 20 months after surgery.
No preview · Article · Aug 2013 · Genetic Testing and Molecular Biomarkers
[Show abstract][Hide abstract] ABSTRACT: Unlabelled:
The allele 1691A F5, conferring Factor V Leiden, is a common risk factor in venous thromboembolism. The frequency distribution for this allele in Western Europe has been well documented; but here data from Central, Eastern and South-Eastern Europe has been included. In order to assess the significance of the collated data, a chi-squared test was applied, and Tukey tests and z-tests with Bonferroni correction were compared.
A distribution with a North-Southeast band of high frequency of the 1691A F5 allele was discovered with a pocket including some Southern Slavic populations with low frequency. European countries/regions can be arbitrarily delimited into low (group 1, <2.8 %, mean 1.9 % 1691A F5 allele) or high (group 2, ≥2.8 %, mean 4.0 %) frequency groups, with many significant differences between groups, but only one intra-group difference (the Tukey test is suggested to be superior to the z-tests).
In Europe a North-Southeast band of 1691A F5 high frequency has been found, clarified by inclusion of data from Central, Eastern and South-Eastern Europe, which surrounds a pocket of low frequency in the Balkans which could possibly be explained by Slavic migration. There seem to be no indications of variation in environmental selection due to geographical location.
Full-text · Article · Aug 2013 · Journal of applied genetics
[Show abstract][Hide abstract] ABSTRACT: BACKGROUND: Atopic dermatitis (AD) is a common inflammatory skin disease. Previous studies have revealed shared genetic determinants among different inflammatory disorders, suggesting that markers associated with immune-related traits might also play a role in AD. OBJECTIVE: We sought to identify novel genetic risk factors for AD. METHODS: We examined the results of all genome-wide association studies from a public repository and selected 318 genetic markers that were significantly associated with any inflammatory trait. These markers were considered candidates and tested for association with AD in a 3-step approach including 7 study populations with 7130 patients with AD and 9253 control subjects. RESULTS: A functional amino acid change in the IL-6 receptor (IL-6R Asp358Ala; rs2228145) was significantly associated with AD (odds ratio [OR], 1.15; P = 5 × 10(-9)). Interestingly, investigation of 2 independent population-based birth cohorts showed that IL-6R 358Ala specifically predisposes to the persistent form of AD (ORpersistent AD = 1.22, P = .0008; ORtransient AD = 1.04, P = .54). This variant determines the balance between the classical membrane-bound versus soluble IL-6R signaling pathways. Carriers of 358Ala had increased serum levels of soluble IL-6R (P = 4 × 10(-14)), with homozygote carriers showing a 2-fold increase. Moreover, we demonstrate that soluble IL-6R levels were higher in patients with AD than in control subjects (46.0 vs 37.8 ng/mL, P = .001). Additional AD risk variants were identified in RAD50, RUNX3, and ERBB3. CONCLUSION: Our study supports the importance of genetic variants influencing inflammation in the etiology of AD. Moreover, we identified a functional genetic variant in IL6R influencing disease prognosis and specifically predisposing to persistent AD.
Full-text · Article · Apr 2013 · The Journal of allergy and clinical immunology
[Show abstract][Hide abstract] ABSTRACT: A-Kinase Anchoring Proteins (AKAPs) coordinate the specificity of protein kinase A signaling by localizing the kinase to subcellular sites. The 1936G (V646) AKAP10 allele has been associated in adults with low cholinergic/vagus nerve sensitivity, shortened PR intervals in ECG recording and in newborns with increased blood pressure and higher cholesterol cord blood concentration. The aim of the study was to answer the question of whether 1936A > G AKAP10 polymorphism is associated with the newborn electrocardiographic variables.
Electrocardiograms were recorded from 114 consecutive healthy Polish newborns (55 females, 59 males), born after 37 gestational weeks to healthy women with uncomplicated pregnancies. All recordings were made between 3(rd) and 7(th) day of life to avoid QT variability. The heart rate per minute and duration of PR, QRS, RR and QT intervals were usually measured. The ECGs were evaluated independently by three observers. At birth, cord blood of neonates was obtained for isolation of genomic DNA.
The distribution of anthropometric and electrocardiographic variables in our cohort approached normality (skewness < 2 for all variables). No significant differences in anthropometric variables and electrocardiographic traits with respect to AKAP10 genotype were found. Multiple regression analysis with adjustment for gender, gestational age and birth mass revealed that QTc interval in GG AKAP10 homozygotes was significantly longer, but in range, when compared with A alleles carriers (AA + AG, recessive mode of inheritance). No rhythm disturbances were observed.
Results demonstrate possible association between AKAP10 1936A > G variant and QTc interval in Polish newborns.
Full-text · Article · Mar 2013 · Archives of Medical Science
[Show abstract][Hide abstract] ABSTRACT: Introduction. Abdominal aorta aneurysm is a recognized cause of death for the European population. The identification of intra-operative risk factors for perioperative death is of crucial importance for the society. Material and methods. A prospective observational study was conducted in the Vascular Surgery Department of the Pomeranian Medical University in Szczecin, Poland. The study group consisted of 95 patients, diagnosed with abdominal aorta aneurysm in the sub-renal region scheduled for an operative procedure of straight vascular graft implantation. Patient qualification was fulfilled according to TASC criteria. The influence of preoperative factors, hemodynamic and metabolic parameters on the risk of death in the study population was analyzed. Postoperatively, observation was continued for 28 days. Results. In this study group an increase of lactate, potassium levels or pH decrease during the first minute post cross-clamp release were significant risk factors for perioperative death. The importance of metabolic parameters is a recognized risk factor, however their statistical significance within the first minute post cross-clamp release is in our opinion of crucial importance. Additionally, significantly higher mortality was reported among patients without epidural anaesthesia. No importance of other analyzed parameters was found. Conclusions. Epidural anaesthesia is an independent factor decreasing mortality in the early postoperative period in patients undergoing abdominal aorta aneurysm repair. A decrease of pH value and increase of lactate and K+ levels within the first minute after aortic cross-clamp release may be a valuable tool in identifying patients with an increased risk of perioperative death after abdominal aorta prosthesis implantation.
No preview · Article · Jan 2013 · Acta Angiologica