The influence of P2Y12 gene polymorphisms on clopidogrel therapy in patients after percutaneous coronary intervention.

Aim: We aimed to define the influence of P2Y12 polymorphisms (rs6801273, rs2046934, and rs6809699), diabetes, hypertension, obesity, hypercholesterolemia, statins intake, and smoking habit on clopidogrel therapy in patients undergoing percutaneous coronary intervention. Materials & methods: We used PCR-RFLP and PCR-ASO for P2Y12 genotype analysis. The effectiveness of the therapy was measured with the VerifyNow method and defined in platelet reactivity units. Results: Studied polymorphisms had no statistically significant influence on PRU before (PRU0) and 6 months (PRU6) after the procedure. H1/H1 diabetic carriers had significantly higher PRU6 values than patients without diabetes. Obese H1/H2 subjects had significantly lower PRU6 values than H1/H2 non-obese carriers. Conclusion: We found that obesity and diabetes may influence the long-term outcome of antiplatelet therapy.

Clopidogrel is a medicine that prevents platelets in the blood from clumping and blocking arteries. When the structure of the protein (e.g., P2Y12), responsible for response to clopidogrel is changed, we can observe less efficient therapy. Said changes can be caused for example by genetic polymorphisms, which are two or more variants of the same gene. This is why we wanted to check the impact of P2Y12 polymorphisms. We also wanted to check the impact of diabetes, high blood pressure, being overweight, high cholesterol blood level, cholesterol-reducing drugs, and smoking habits on clopidogrel treatment in patients after a procedure that unblocks blood vessels of the heart to restore its blood supply (percutaneous coronary intervention). We measured the efficacy of the treatment with platelet reactivity units (PRU). Studying polymorphisms had no impact on treatment efficacy before (PRU0) and 6 months (PRU6) after the medical procedure. We found that diabetes can cause higher platelet reactivity after 6 months of therapy. We noticed that being overweight may also be important, as obese patients had lower platelet reactivity values.

The Expression and Prognostic Significance of VEGF and CXCR4 in Gastric Cancer: Correlation with Angiogenesis, Lymphangiogenesis and Progression.

The cellular response to hypoxia includes the expression of hypoxia-inducible factor-1 (HIF-1) and its target genes: vascular endothelial growth factor (VEGF) and CXC chemokine receptor 4 (CXCR4). The aim of this study was to investigate the expression and prognostic significance of VEGF and CXCR4, which are responsible for angiogenesis and progression in gastric cancer. Twenty-eight gastric cancer patients were analyzed. The mRNA expression was examined in primary tumors and corresponding normal gastric mucosa by RT-PCR. The protein level was examined by immunohistochemistry staining. The high expression of VEGF and CXCR4 was found in 71.0 and 64.0% of tumors, respectively. The mean levels of VEGF and CXCR4 were upregulated in primary tumors compared to normal mucosa (p = 0.0007, p = 0.0052). A correlation between VEGF expression and tumor invasion (p = 0.0216) and stage (p = 0.0181) was found. CXCR4 expression correlated with lymph node metastases (p = 0.0237) and stage (p = 0.0054). The VEGF expression correlated with microvessel density (MVD) (p = 0.0491). The overall 3-year survival rate was 46.4% and correlated negatively with high CXCR4 mRNA expression (p = 0.0089). VEGF and CXCR4 play an important role in tumor progression. Their overexpression correlates with a bad prognosis and may improve high-risk patient selection, and these patients may obtain additional survival benefits if treated more aggressively.

The influence of non-thrombotic iliac vein stenosis on clinical course and recurrence of primary varicose veins.

INTRODUCTION: To investigate the influence of iliac vein stenosis on clinical course and recurrence of primary varicose veins after surgeryMaterials and methods: Thirty-three patients with primary varicose veins qualified for great saphenous vein stripping were analysed. The stenosis of common (CIV) and external (EIV) iliac vein was measured by IVUS and defined in three categories as minimal lumen area <90 mm2 for CIV and <75 mm2 for EIV, minimal lumen diameter <10 mm for CIV and <7.5 mm for EIV and area reduction >50%. The patients were assessed clinically and by Duplex ultrasound 48 to 72 months after the procedure. Any recurrence, the recurrence in the saphenofemoral junction (SFJ), change in Venous Clinical Severity Score ( VCSS), were analyzed in relation to the stenosis in the CIV and EIV. RESULTS: The follow-up was completed in 27 patients. Any recurrence and the recurrence in the SFJ were observed in 70% and 18.5% of patients, respectively. There were no statistically significant differences in any recurrence, the recurrence in the SFJ and VCSS in relation to CIV and EIV stenosis in any category. CONCLUSIONS: Iliac vein stenosis does not influence the clinical course and recurrence of primary varicose veins after surgery.

Midterm Results of the Treatment of Penetrating Abdominal Aortic or Iliac Artery Ulcer with the BeGraft Balloon-Expandable Covered Stent-A Single-Center Experience.

BACKGROUND: The purpose of the study was to evaluate the feasibility and efficacy of the novel BeGraft covered stent for the treatment of abdominal penetrating aortic ulcer (PAU) or penetrating ulcers of the iliac arteries (PUIAs). METHODS: This was a single-center observational study, which included 24 consecutive patients who underwent endovascular surgery due to abdominal PAU or PUIA between June 2017 and September 2019. Demographics of patients, lesion characteristics, diameter and length of the BeGraft stents, and postoperative events were prospectively collected and retrospectively analyzed. Follow-up examinations were performed at 1, 6, 12, and 24 months with clinical and hemodynamic evaluation. Outcome measures included technical success, perioperative complications, and stent patency. RESULTS: A total of 24 patients (13 men and 11 women), with a median age of 67 years (range, 42-81 years), were analyzed. Among them, 20 patients were symptomatic, and 4 patients underwent elective surgery because of the size of PAU. A total of 54 BeGraft stents (26 aortic and 28 peripheral) were successfully delivered and deployed to cover 13 aortic and 13 common iliac artery ulcer lesions. The technical success rate was 100%. The average procedural time was 53.8 ± 12.8 min. Complications included one case of the access-site pseudoaneurysm, which was successfully treated by thrombin injection. During a median follow-up of 20.5 months (range, 6-33 months), all stents remained patent, without endoleak or ulcer recurrence. CONCLUSIONS: BeGraft stents used during endovascular treatment of abdominal PAU and PUIA lesions are associated with favorable outcomes regarding technical success and patency. The primary use of BeGraft covered stents provides a valid option for patients with abdominal PAU. Long-term follow-up is required to confirm these promising results.

The Proliferation and Differentiation of Adipose-Derived Stem Cells in Neovascularization and Angiogenesis.

Neovascularization and angiogenesis are vital processes in the repair of damaged tissue, creating new blood vessel networks and increasing oxygen and nutrient supply for regeneration. The importance of Adipose-derived Mesenchymal Stem Cells (ASCs) contained in the adipose tissue surrounding blood vessel networks to these processes remains unknown and the exact mechanisms responsible for directing adipogenic cell fate remain to be discovered. As adipose tissue contains a heterogenous population of partially differentiated cells of adipocyte lineage; tissue repair, angiogenesis and neovascularization may be closely linked to the function of ASCs in a complex relationship. This review aims to investigate the link between ASCs and angiogenesis/neovascularization, with references to current studies. The molecular mechanisms of these processes, as well as ASC differentiation and proliferation are described in detail. ASCs may differentiate into endothelial cells during neovascularization; however, recent clinical trials have suggested that ASCs may also stimulate angiogenesis and neovascularization indirectly through the release of paracrine factors.

The intravascular ultrasound morphometry of iliac veins in subjects without severe chronic venous insufficiency and its implications for treatment indications and stent size selection.

OBJECTIVES: The purpose of this study is to report the intravascular ultrasound morphometry of iliac veins and its relation to demographic and anthropometric factors in subjects without chronic venous insufficiency. METHODS: Thirty-three patients, without chronic venous insufficiency - qualified to great saphenous vein stripping due to unilateral, primary varicose veins - participated in the study. During the surgery, left and right external iliac veins, common iliac veins and inferior vena cava were interrogated with intravascular ultrasound. The morphometric analysis included measurement of a cross-sectional area at normal, non-stenosed vein segments (ref-CSA) and at the point of the most prominent narrowing (minimal lumen area (MLA)). Based on these measurements, a percentage of stenosis (S%) and calculated lumen diameter of interrogated veins were determined according to the following formulas, S% = (ref-CSA-MLA)/ref-CSA × 100 and CLD = 2 × âˆš(ref-CSA/π), respectively. RESULTS: Median ref-CSA, S% and calculated lumen diameter were 265.3 mm2, 45.8% and 18.4 mm for inferior vena cava; 193.9 mm2, 62.4% and 15.7 mm for left common iliac veins; 166.9 mm2, 35.7% and 14.2 mm for right common iliac veins; 136.5 mm2, 48.0% and 12.8 mm for left external iliac veins and 140.9 mm2, 46.3% and 13.5 mm for right external iliac veins. There were statistically significant differences between left and right common iliac veins ref-CSA, common iliac veins S% and common iliac veins calculated lumen diameter (p = 0.03, p < 0 and p = 0.03, respectively). The S% of left external iliac veins was greater in women 52.2 versus 37.2% in men (p = 0.04). Neither age nor anthropometric factors had any influence on the calculated lumen diameter of the analysed veins. A negative correlation between the left common iliac veins S% and the age was observed (p = 0.03). CONCLUSIONS: In adult subjects, the calculated lumen diameter of the common iliac veins is greater on the left side and is not influenced by age and body size. Common iliac vein stenosis occurs more frequently on the left side, decreases with age and tends to be more frequent in women.

Alterations in Exercise-Induced Plasma Adenosine Triphosphate Concentration in Highly Trained Athletes in a One-Year Training Cycle.

This study aimed to assess the effect of training loads on plasma adenosine triphosphate responsiveness in highly trained athletes in a 1 y cycle. Highly trained futsal players (11 men, age range 20-31 y), endurance athletes (11 men, age range 18-31 y), sprinters (11 men, age range 21-30 y), and control group (11 men, age range 22-34 y) were examined across four characteristic training phases in response to an incremental treadmill test until exhaustion. A considerably higher exercise and post-exercise plasma adenosine triphosphate concentrations were observed in consecutive training phases in highly trained athletes, with the highest values reached after the competitive period. No differences in plasma adenosine triphosphate concentrations were found in the control group during the 1 y cycle. Sprinters showed a higher absolute and net increase in plasma adenosine triphosphate concentration by 60-114% during exercise in consecutive training phases than futsal players (63-101%) and endurance athletes (64-95%). In this study, we demonstrated that exercise-induced adenosine triphosphate concentration significantly changes in highly trained athletes over an annual training cycle. The obtained results showed that high-intensity but not low- to moderate-intensity training leads to an increased adenosine triphosphate response to exercise, suggesting an important role of ATP for vascular plasticity.

Clinical pharmacokinetics of clopidogrel and its metabolites in patients with cardiovascular diseases.

BACKGROUND AND OBJECTIVE: Approximately 5-40 % of patients treated with clopidogrel do not display an adequate antiplatelet response. Clopidogrel resistance may be caused by insufficient drug absorption or impaired metabolic activation of the drug. The aim of this study was to evaluate the pharmacokinetics of clopidogrel and its metabolites in plasma samples from patients treated with high and low doses of clopidogrel, to obtain a possible explanation for antiplatelet resistance. METHODS: The study included patients receiving either a single 300 mg loading dose of clopidogrel (n = 17) or a 75 mg dose (n = 45) for at least 7 days before sample collection. The concentrations of clopidogrel and its metabolites-the inactive H3 and the pharmacologically active H4 isomers of the thiol metabolite and the inactive carboxylic acid metabolite-in plasma samples (stabilized with 2-bromo-3'-methoxyacetophenone) from three patients after 300 mg and from 41 patients after 75 mg of the drug were determined using a validated high-performance liquid chromatography method with tandem mass spectrometry. The non-stabilized samples from the remaining patients were analysed using a validated capillary electrophoresis method. The calculated concentrations were used to determine the pharmacokinetic parameters of the analytes. The pharmacodynamic response to clopidogrel treatment, expressed as adenosine diphosphate-induced platelet aggregation, was measured using a Multiplate analyser. RESULTS: The pharmacokinetic parameter values for the H3 and H4 isomers determined in the studied group of patients treated with clopidogrel 75 mg (maximum plasma concentration [C max] 5.29 ± 5.54 and 7.13 ± 6.32 ng/mL for H3 and H4, respectively; area under the plasma concentration-time curve from time zero to time t [AUC t ] 7.37 ± 6.71 and 11.30 ± 9.58 ng·h/mL for H3 and H4, respectively) were lower than those reported in healthy volunteers, according to the literature data. Platelet aggregation measured with a Multiplate analyser ranged between 37 and 747 AU·min. A significant correlation was found between the C max of the active H4 isomer and platelet aggregation (p = 0.025). CONCLUSION: The C max of the active H4 isomer and platelet aggregation measured by the Multiplate analyser may serve as markers of the patient response to clopidogrel therapy.

Assessment of the level of difficulty of four techniques of endovenous thermal ablation of the great saphenous vein and the echogenicity of the tip of the working device in vivo.

BACKGROUND: To compare the level of difficulty of four techniques of endovenous thermal ablation (EVTA) of the great saphenous vein and the echogenicity of the tip of the working device in vivo. METHODS: Sixty patients qualified to the EVTA of the great saphenous vein were randomly assigned to treatment with an 810-nm axial diode laser [endovenous laser ablation (EVLA) 810] with two different delivery systems: 4-F introducer, 0.018" guidewire, 22-G needle (EVLA810-1) and 4-F introducer, 0.035" guidewire, 19-G needle (EVLA810-2); a 1470-nm radial diode laser (EVLA1470); or radiofrequency ablation (RFA; ClosureFAST). The level of difficulty of four stages of the procedure-cannulation of a vein, advancement of the working part to the saphenofemoral junction (SFJ), visualization of a tip of the working part at SFJ, and difficulty of performing the ablation and delivering the planned linear energy density-was subjectively assessed. An objective comparison of visibility of working parts in ultrasonography was performed with analysis of grayscale median. RESULTS: The cannulation of a distal segment of the obliterated vein was the most difficult in EVLA810-1, P = 0.015. The delivery of a working part to the SFJ was the least problematic in RFA and EVLA1470, P = 0.024. The visualization of the working tip at the SFJ was the most difficult in RFA, P = 0.028. The application of desired amount of energy was the easiest in RFA, P = 0.038. The EVLA1470 presented the best echogenicity. CONCLUSIONS: Although all the examined techniques have advantages and disadvantages, EVTA with the 1470-nm diode laser with radial optic fiber seems to be the easiest.

CCL2 -2518 A/G single nucleotide polymorphism as a risk factor for breast cancer.

The contribution of the CCL2 -2518 A>G (rs 1024611) polymorphism in the occurrence and progression of various cancers has been found to be discordant. We studied the prevalence of the CCL2 -2518 A>G polymorphism in patients with breast cancer (n = 160) and controls (n = 323) in a sample of the Polish population. There were no significant differences in CCL2 -2518 A>G genotypes between patients with breast tumors and controls. Odds ratio (OR) for patients bearing the GG genotype was 1.481 (95% CI = 0.7711-2.845, P = 0.2358), and OR of the GG and AG genotypes was 0.7269 (95% CI = 0.4967-1.064, P = 0.1002). There was also no significant distinction in the prevalence of alleles between patients and healthy individuals. OR for the CCL2 -2518 G allele frequency was 0.8903 (95% CI = 0.6611-1.199, P = 0.4441). Analysis of the association between tumor size, lymph node metastases, histological grade, and distribution of genotypes and alleles for the CCL2 -2518 A>G polymorphism also did not show significant differences. Our results did not show association of the CCL2 -2518 A>G polymorphism with breast cancer occurrence and clinical characteristics in a sample of the Polish cohort.

Distribution of CCND1 A870G polymorphism in patients with advanced uterine cervical carcinoma.

We examined the distribution of the CCND1 A870G (rs9344) polymorphic variant in patients with cervical cancer (n = 129) and healthy individuals (n = 288) in a sample of a Polish cohort. We showed that patients with advanced cervical cancer bearing the CCND1 A/A and A/G genotypes displayed a 1.811-fold increased risk of cervical cancer (95% CI = 1.150-2.852, p = 0.0098). We also found a significantly higher frequency of the CCND1 870A allele in patients with cancer than in controls, p = 0.0116. Our investigation confirmed that the CCND1 870A gene variant may be a genetic risk factor in the incidence of advanced cervical cancer.

CXCL12-3' G801A polymorphism is not a risk factor for breast cancer.

The CXCL12-3' G801A transition (rs1801157) has been associated with the incidence of breast cancer. However, the contribution of CXCL12-3' G801A polymorphisms in breast cancer development and progression has been controversial. Therefore, we examined the incidence of CXCL12-3' G801A polymorphic variants in patients with breast cancer (n = 193) and controls (n = 199) in a Polish cohort. We observed a trend of slightly increased presence of CXCL12-3' AA and GA genotypes and CXCL12-3'A allele frequency in patients with breast cancer compared with healthy individuals. However, these differences between cases and controls were not statistically significant. Odds ratio (OR) for patients with breast cancer and the CXCL12-3' A/A genotype was 1.898 (95% confidence interval [CI] = 0.6242-5.770, p = 0.2866) and OR of the CXCL12-3' A/A and A/G genotypes was 1.229 (95% CI = 0.8082-1.868, p = 0.3395). OR for the CXCL12-3'A allele frequency was 1.249 (95% CI = 0.8716-1.789, p = 0.2352). Our investigation did not support the CXCL12-3'A gene variant as a risk factor for breast cancer incidence in a sample of the Polish population.

HER2 codon 655 polymorphism is associated with advanced uterine cervical carcinoma.

OBJECTIVES: It has been suggested that overexpression of HER2 in advanced cervical tumors can be considered an independent predictor of poor patient outcome. DESIGN AND METHODS: Employing PCR-RFLPs, we examined the distribution of HER2 Ile655Val (rs 1136201) genotypes and alleles in patients with advanced cervical cancer (n=109) and controls (n=220). RESULTS: Odds ratio (OR) for patients with advanced cervical cancer with the HER2 Val/Val homozygous or Val/Ile heterozygous state was 1.778 (95% CI=1.117-2.830, p=0.0176). We also observed an association of the HER2 Val/Val genotype with advanced cervical cancer in the patient group OR=3.706 (95% CI=1.061-12.950, p=0.0459). However, we did not find a significant association between the distribution of genotypes or alleles and cancer characteristics for the HER2 Ile655Val polymorphism. CONCLUSIONS: Our results indicate that the HER2 655Val variant may be associated with the incidence of advanced cervical cancer.

Polymorphic variants of folate metabolism genes and the risk of laryngeal cancer.

Carcinogenesis may result from abnormal methylation of cancer-related genes regulatory sequence. Though, the polymorphic variants of genes encoding enzymes of folate and methionine metabolism may have an effect on DNA methylation. Using PCR-RFLPs, we examined the polymorphism distribution of genes encoding methionine synthase (MTR); 5,10-methylenetetrahydrofolate dehydrogenase, 5,10-methenyltetrahydrofolate cyclohydrolase and 10-formyltetrahydrofolate synthetase (MTHFD1); and methylenetetrahydrofolate reductase (MTHFR) in patients with larynx cancer (n = 131) and controls (n = 250). Patients with MTR 2756AG or GG genotypes displayed a 1.856 -fold increased risk of larynx cancer (95% CI = 1.1860-2.903, P = 0.0076). However, we did not observe an increased risk for the homozygous GG genotype OR = 1.960 (95% CI = 0.6722-5.713, P = 0.2535). Moreover, we did not observe statistical differences in distribution of MTHFR 677C>T, 1298A>C and MTHFD1 1958G>A allele and genotype frequencies in patients and controls. Our findings confirm the significance of the role of the methyl cycle in etiopathogenesis of laryngeal cancer.

SDF1-3' a gene polymorphism is associated with laryngeal cancer.

The SDF1-3' G801A (rs 1801157) polymorphism is associated with increased risk of various types of cancers, including those of the neck and head. Using PCR-RFLPs, we investigated the distribution of SDF1-3' G801A genotypes in patients with laryngeal cancer (n = 118) and controls (n = 250) in Poland. We found that patients with SDF1-3' A/A and G/A genotypes exhibit a 1.863-fold increased risk of laryngeal cancer (95% CI = 1.177-2.949, p = 0.0086). However, there was no significant increase in risk for the homozygous SDF1-3' A/A genotype OR = 3.235 (95% CI = 0.5330-19.633, p = 0.3329). We also did not observe a significant association between tumor characteristics and prevalence of alleles or genotypes for the SDF1-3' G801A polymorphism. Our findings suggest that the SDF1-3'A variant may be associated with an increased risk of laryngeal cancer.

[Mutations of tumor suppressor gemne TP53 in tobacco smoke-associated tumors]. / Mutacje supresorowego genu przeciwnowotworowego TP53w nowotworach tytonio-zaleznych.

Tumour suppressor gene TP53 is a subject of frequent lesions and mutations in a majority of cancer types that is followed by its dysfunction in regulation of cell proliferation, apoptosin and DNA repair. Mutation profile reflects the presence of mutagen-vulnerable sites (including tobacco smoke carcinogens) in its structure. A number of mutations in tobacco smoke-associated cancers are higher than in other types. Particularly, G > T mutation is recognized a signature to benzo(a)pyrene exposure. Further, a mutation profile is dependent on cancer anatomic localization and histological type. There were put forward suggestions concerning estimation of cancer risk and disease prognosis basing of TP53 gene status and expression. The protocols of gene therapy involving TP53 gene are still not satisfactory.

Mutations within protein kinase R-binding domain of NS5A protein of hepatitis C virus (HCV) and specificity of HCV antibodies in pretreatment sera of HCV-chronically infected patients and their effect on the result of treatment.

We analyzed protein kinase R (PKR)-binding domain sequences of hepatitis C virus (HCV) NS5A protein and the profile of HCV-specific antibodies from pretreatment sera of HCV-chronically infected patients. Results were compared with clinical data to verify their influence on the course and result of therapy. Of 9 patients enrolled in a 12-month treatment with pegylated interferon alpha (PEG-IFN-alpha) plus ribavirin (RBV), 6 patients responded to therapy, as assessed by the lack of HCV RNA in their sera, and 3 did not. Among 8 HCV-1b-infected patients, those who responded did not have significantly more mutations in the IFN sensitivity determining region (ISDR) compared to non-responders (P = 0.637). Similarly, in the remaining 26-amino acid region of the PKR-binding domain, behind ISDR, the number of mutations did not differ significantly between the two groups (P = 0.796). A correlation was found between the presence of envelope 2 (E2)-specific antibodies and the result of treatment (P = 0.048). This pilot study indicates that mutations in the PKR-binding domain of HCV genotype 1b do not correlate with outcome of PEG-IFN-alpha/RBV therapy. However, the presence of E2-specific antibodies in the pretreatment sera of HCV-chronically infected individuals could serve as a prognostic marker predicting the result of treatment, before its initiation.