Robert Milewski

Prognostic significance of DAPK and RASSF1A promoter hypermethylation in non-small cell lung cancer [NSCLC]

The epigenetic inactivation of tumor suppressor genes may play an important role in the development and progression of many cancer types, including lung cancer. Therefore, we investigated the association between the aberrant promoter methylation of 2 genes: the Death-Associated Protein Kinase (DAPK) and the Ras Association Domain Family 1A (RASSF1A) by using methylation-specific PCR, and the clinicopathological features and prognosis in 70 radically resected non-small cell lung cancers (NSCLCs). Hypermethylation of the DAPK and RASSF1A promoters was found in 24 (34%), and in 18 (26%) tumor DNA samples, respectively. Regarding different clinicopathological features of NSCLCs, the DAPK promoter methylation was more frequently observed in squamous cell carcinoma (46%) than in adenocarcinoma (25%) and large cell carcinoma (22%), but there were no significant statistical differences (p=0.3). On the other hand, a statistically significant trend was observed between the RASSF1A methylation and a histological type of tumor (p=0.06). 45% of adenocarcinoma tumors showed RASSF1A promoter methylation in comparison to 17% of squamous cell carcinomas and 22% of large cell carcinomas. When both markers were analyzed according to the tumor-node-metastasis (TNM) staging system, no statistically significant differences were observed between stage I, II and IIIa, and the DAPK (p=0.2) and RASSF1A methylation (p=0.1). In comparison, when stage I and II were grouped together and considered vs. stage IIIa, a significant association between RASSF1A methylation and the TNM was found (p=0.03). The group of patients with tumors showing DAPK promoter methylation had significantly poorer overall survival rates (p=0.02) than the patients with tumors that did not show DAPK promoter methylation. However, the association between the RASSF1A promoter methylation status and the overall survival rates was not statistically significant (p=0.48). In conclusion, this paper supports the importance of epigenetic gene regulation in lung cancer progression and prognosis.

Serum vascular endothelial growth factors C and D in patients with oesophageal cancer.

OBJECTIVE Lymph node metastasis is a characteristic of malignant cancers and is observed more frequently in oesophageal cancer than in other digestive tract cancers, making it one of the most important prognostic factors. Vascular endothelial growth factors C (VEGF-C) and D (VEGF-D) are important lymphangiogenic factors in human cancers and lymphangiogenesis is associated with lymph node metastasis. The aim of the study was to determine the correlation between pre-treatment serum levels of VEGF-C (sVEGF-C) and VEGF-D (sVEGF-D) and clinicopathologic features in patients with oesophageal cancer. METHODS Serum VEGF-C and sVEGF-D were measured by enzyme-linked immunoadsorbent assay (ELISA) on 149 patients with oesophageal cancer, 29 patients with benign oesophageal diseases and 30 healthy controls. RESULTS Serum VEGF-C and sVEGF-D levels were significantly higher in patients with oesophageal carcinoma than in the control group (p<0.001 and p=0.001, respectively) or in the benign oesophageal diseases group (p=0.04 and p=0.03, respectively). Subgroup analysis showed that lymph node metastasis (p=0.001), stage (p=0.001), tumour depth (p=0.006), resectability (p=0.002), tumour size (p=0.01), distant metastases (p=0.01) and histological grading (p=0.04) were correlated with an elevated level of sVEGF-C. Elevated levels of sVEGF-D were associated with tumour depth (p=0.002), stage (p=0.01) and lymph node metastasis (p=0.02). Among the patients (n=83) who underwent potentially curative surgery, the overall survival time (p=0.008) was shorter for patients with a high level (>8667 pg ml(-1)) of sVEGF-C than for those with a low level (<8667 pg ml(-1)), when the cut-off value was determined on the basis of the median value in oesophageal cancer patients. On univariate regression analysis, tumour size, tumour depth, stage, lymph node metastases, distant metastases, resectability and sVEGF-C were found to be significant prognostic factors. CONCLUSIONS These results suggest that pre-treatment levels of sVEGF-C and sVEGF-D reflect lymph node metastases and advanced stage of oesophageal cancer. Serum VEGF-C may be useful in predicting poor outcome for patients undergoing a potentially curative oesophagectomy.

Proton magnetic resonance spectroscopy measures related to short-term symptomatic outcome in chronic schizophrenia.

INTRODUCTION Proton magnetic resonance spectroscopy (¹H MRS) enables the evaluation of in vivo brain function. The purpose of the study was to compare ¹H MRS measurements in schizophrenic patients, who were clinical responders after short-term antipsychotic treatment, with non-responders and healthy controls. METHODS We investigated a group of 47 patients diagnosed with schizophrenia. Patients were examined twice--once after a period of at least 7 days without neuroleptics and the second time at least 4 weeks after therapy with stable doses of medication. The follow-up was available in 42 patients. Baseline MRS measurements of clinical responders were compared with non-responders and the group of healthy controls (N=26). We assessed the following metabolite ratios: NAA (N-acetylaspartate), Glx (complex of GABA, glutamine and glutamate), Cho (choline) and mI (myo-inositol) to creatinine (Cr) in the left frontal and temporal lobes and the thalamus. RESULTS Responders showed a significantly lower baseline frontal Glx/Cr level than non-responders. Both groups had a significantly lower NAA/Cr ratio in the frontal lobe than the controls, but only non-responders had a significantly lower NAA/Cr ratio in the thalamus. CONCLUSIONS Our results confirm the relationship between the glutamatergic system and pathophysiology of schizophrenia and suggest a significant value of ¹H MRS examination in the assessment of the future treatment effect.

Evaluation of the influence of ozonotherapy on the clinical parameters and MMP levels in patients with chronic and aggressive periodontitis

PURPOSE A comparison of the clinical status and salivary MMP levels after SRP alone or with ozonotherapy in patients with aggressive and chronic periodontitis. MATERIAL/METHODS The study was performed in 52 generally healthy subjects with chronic or aggressive periodontitis. Group CP-S consisted of 12 patients with chronic periodontitis, who underwent scaling and root planing (SRP). In group CP-O there were 25 patients with chronic periodontitis who additionaly to SRP underwent ozonotherapy. The same therapy was performed in group AP, containing 15 patients with aggressive periodontitis. Plaque index, approximal plaque index, bleeding on probing, sulcus bleeding index, probing pocket depth and clinical attachment loss were measured at baseline, at two weeks and two months post-therapy. The levels of MMP-1, MMP-8 and MMP-9 were estimated in non-stimulated saliva with an ELISA method. RESULTS All the clinical parameters assessed in the study groups were reduced after treatment. SRP with additional ozonotherapy provided an increase in MMP levels in patients with chronic periodontitis and a reduction in MMP levels in patients with aggressive periodontitis. CONCLUSIONS SRP followed by ozonotherapy does not lead to further improvement in clinical periodontal parameters in patients with AP and CP.

Urinary angiotensinogen as a marker of intrarenal angiotensin II activity in adolescents with primary hypertension

BackgroundExperimental and epidemiological studies have demonstrated that urinary angiotensinogen (AGT) is a novel biomarker for the intrarenal activity of the renin–angiotensin system in hypertension (HT). Several large-scale epidemiological studies have shown that an elevated serum uric acid (SUA) level is associated with HT. The aim of our study was to assess urinary AGT excretion and its correlation with SUA level, the lipid profile, and the body mass index (BMI) Z-score in hypertensive adolescents.MethodsParticipants were divided into two groups: (1) the group with confirmed HT consisting of 55 subjects with primary HT and (2) the reference (R) group consisting of 33 subjects with white-coat HT. A commercial enzyme-linked immunosorbent assay (ELISA) kit was used to determine urinary AGT concentration.ResultsThe urinary AGT/creatinine (cr.) ratio in subjects in the HT group was significantly higher than that in the reference group (p < 0.01) and showed a strong positive correlation with SUA (r = 0.47, p < 0.01). The relationship between the AGT/cr. ratio and SUA levels after controlling for age, gender and BMI Z-score continued to show a significant association.ConclusionsThe most obvious finding to emerge from this study is that in adolescents with primary HT, the increased urinary excretion of AGT correlated with hyperuricemia, although large, multicenter studies are needed to confirm this observation.

Thrombopoiesis in small for gestational age newborns.

Thrombocytopenia in small for gestational age (SGA) newborns may be due to placental vascular pathology, fetal consumptive coagulopathy and platelet destruction, local imbalance of thromboxane A2 causing placental vasoconstriction and platelet aggregation. Thrombopoiesis in SGA newborns is poorly recognized. In 61 SGA newborns we evaluated thrombocytopoiesis in relation to gender and the rate maturity expressed as <5th percentile and <10th percentile. Female newborns demonstrated higher thrombopoietin (TPO) level at 92.06 pg/ml than male newborns at 79.81 pg/ml. Newborns less developed <5th percentile, showed increased TPO level of 92.0 pg/ml in comparison to <10th percentile of 78.0 pg/ml. This observation is more pronounced in female newborns. Contrary to our expectations we did not find any statistically significant differences in the percentage of reticulated platelets (PLRET) and platelets count in relation to gender and <5th percentile or <10th percentile. We can postulate intrauterine hypoxia is responsible for the increase of erythropoietin and impairment of thrombopoiesis in SGA newborns.

Lymphatic vessel invasion detected by the endothelial lymphatic marker D2-40 (podoplanin) is predictive of regional lymph node status and an independent prognostic factor in patients with resected esophageal cancer.

The discovery of markers to lymphatic endothelial cells and the development of novel antibodies to these markers have brought increasing attention to the lymphatics and progress in the understanding of lymphangiogenesis and cancer metastasis. In this study, we investigate the presence of lymphatic vessel invasion (LVI) detected by D2-40 immunohistochemical staining in resected esophageal cancer and correlated with clinicopathologic data and patient survival. Sixty nine patients, who had a primary resection of esophageal cancer, were analyzed by univariate and multivariate logistic regression, and univariate and multivariate survival analysis. The total rate of LVI was 72% (50/69). Positive LVI was significantly correlated with lymph node metastasis (p < 0.001), tumor size (p < 0.001), histological grading (p = 0.017), tumor depth (p = 0.001), and stage (p < 0.001). Multivariate logistic analysis identified LVI (p = 0.036) as a predictor of regional lymph node metastasis. On univariate survival analysis, patients with LVI had a significantly shorter disease-free survival, cancer-specific survival and overall survival. Multivariate analysis proved that LVI diagnosed by D2-40 is an independent prognostic factor of both disease-free survival (p = 0.04) and overall survival (p = 0.032) in resected esophageal cancer. These results show that LVI assessment identifies patients at high risk for regional lymph node metastasis and that LVI is an independent prognostic factor in patients with esophageal cancer.

Changes in blood eosinophilia during omalizumab therapy as a predictor of asthma exacerbation.

Introduction Omalizumab is a monoclonal anti-immunoglobulin E antibody developed for the treatment of severe allergic asthma. The number of exacerbations used as a parameter of omalizumab therapy efficacy may be insufficient in many cases due to a relatively short time to first evaluation (16 weeks). Therefore, it is advisable to look for parameters of more prognostic value while continuing omalizumab therapy. Aim To evaluate usefulness of analysis of changes of blood eosinophilia after 16 weeks of omalizumab therapy as a predictor of asthma exacerbations. Material and methods The study was conducted on a group of 13 patients with severe persistent allergic asthma treated with omalizumab. Blood eosinophil counts were measured before and after 16 weeks of anti-IgE therapy. On the basis of percentage of eosinophilia decrease (> 50% or < 50% of the initial value), patients were divided into two groups. Analysis of the asthma exacerbation rate during 12 months and time to first exacerbation was performed. Results In the group with a high decrease in blood eosinophil counts (group 1) we showed a statistically significantly lower asthma exacerbation rate in 12 months compared with the group with a low decrease in blood eosinophil counts (group 2) (p = 0.02). We also observed the tendency to longer time to first asthma exacerbation in group 1 compared to group 2 (p = 0.06). Conclusions Our results showed that a decrease in blood eosinophilia during omalizumab therapy can be a predictor of asthma exacerbation. Evaluation of changes in blood eosinophil count should be taken into the consideration while estimating response to anti-IgE therapy in patients with severe allergic asthma.

Polymorphism of 9p21.3 Locus Is Associated with 5-Year Survival in High-Risk Patients with Myocardial Infarction

Objective The rs1333049, rs10757278 and rs4977574 are single nucleotide polymorphisms (SNPs) of chromosome 9p21 locus that are associated with prevalence of acute coronary syndromes (ACS). The rs1333049 SNP was also associated with cardiac outcome 6 months post ACS. No data concerning their association with long term prognosis after myocardial infarction is available. The aim of our study was to investigate the association of the 9p21.3 locus with 5-year overall mortality in patients with ST-elevation myocardial infarction (STEMI) treated invasively. Materials and Methods We performed a retrospective analysis of data collected prospectively in a registry of consecutive patients with STEMI treated with primary PCI. Genotyping was performed with a TaqMan method. The analyzed end-point was total 5-year mortality. Results The study group comprised 589 patients: 25.3% of females (n = 149), mean age 62.4±11.9 years, total 5-year mortality 16.6% (n = 98). When all the study group was analyzed, no significant differences in mortality were found between the genotypes. However, in high-risk patients (Grace risk score ≥155 points, n = 238), low-risk homozygotes had significantly better 5-year survival compared to other genotypes. The hazard ratio associated with high-risk genotype (high-risk homozygote or heterozygote) was: HR = 2.9 (95%CI 1.4–6.1) for the rs4977574 polymorphism, HR = 2.6 (1.25–5.3) for the rs1333049 one and HR = 2.35 (1.2–4.6) for the rs10757278 one (Cox proportional hazards model). Conclusions The 9p21.3 locus is associated with 5-year mortality in high-risk patients with STEMI. This finding, due to very high effect size, could potentially be applied into clinical practice, if appropriate methods are elaborated.

Dental caries in primary and permanent molars in 7-8-year-old schoolchildren evaluated with Caries Assessment Spectrum and Treatment (CAST) index

BackgroundNo reports on a caries pattern covering the full spectrum of the disease could be found in the literature. The aim of this study was to evaluate caries in primary and first permanent molars of 7-8-year-old Polish children by the Caries Assessment Spectrum and Treatment (CAST) index and to find whether there was any correlation between the caries stages in such teeth.MethodsThe study covered 284 7-8-year-old children from randomly selected schools in the Bialystok District, Poland. The prevalence of CAST categories was evaluated with regard to the first and second primary, and first permanent, molars. The Spearman’s rank correlation coefficient was used to explore the correlation of the distribution of CAST codes among the evaluated teeth. The level of statistical significance was established at p < 0.05. The intra-examiner reliability was determined by the unweighted kappa coefficient.ResultsWith regard to the permanent molars, caries was observed in 14.8% to 17.3% of the molar and most lesions were scored at the non-cavitation level. Caries in primary molars was most often recorded at the stage of cavitated dentine lesion. Teeth with pulpal involvement, sepsis and extracted due to caries were found to be more prevalent in first, and then in second primary molars. A strong correlation was found between the status of teeth from the right and left sides of the oral cavity. The correlation of the status of first and second primary teeth was stronger for the left than for the right side of the mouth, r was 0.627 and 0.472 in maxilla and 0.513 and 0.483 in mandible (p < 0.001), respectively. For the neighbouring primary and permanent molars the correlation was assessed to be weak. With regard to the teeth situated in opposite jaws the study revealed that the correlations were moderate - r between 0.33 and 0.49. The intra-examiner reliability was established at 0.96 for the primary dentition and at 0.878 for permanent molars.ConclusionThe strongest correlation found in the evaluated population concerned the distribution of caries in primary molars on the left side of the mouth. The study proved the usefulness of the CAST index in epidemiological surveys.