Prognostic value of tumour tissue ANG-1 expression and Ang-1 concentration in patients with non-small-cell lung cancer.

Tumor cells stimulate local angiogenesis, resulting in their further multiplication and spread. Angiogenesis is a multifaceted process in which angiopoietins parti- cipate. Angiopoietin-1 (Ang-1) through its receptor Tie2 stimulates endothelial cell survival and the maintenance of the endothelial barrier. These phenomena can support tumour growth by promoting angiogenesis. On the other hand, overproduction of Ang-1 triggers endothelium stability and can lead to angiogenesis inhibition. Because of the ambiguous role of Ang-1, we decided to determine its clinical significance in patients with resectable NSCLC. In a group of 47 patients, tumours and the adjacent non-cancerous tissues were assessed for ANG-1 mRNA expression (using Q-RT-PCR analysis) and Ang-1 concentration (by enzyme-linked immunosorbent assay) together with clinical parameters and the five-year survival rate. ANG-1 expression and Ang-1 concentration were higher in tumour-free tissue, showing no differences between histological types of NSCLC, clinical stage or grading and seemed not to determine the five-year survival. ANG-1 expression and Ang-1 concentration in tumour and tumour-free tissues in patients with NSCLC seem not to be useful as factors supporting either diagnostics or prognosis.

Activity of Antioxidant Enzymes in the Tumor and Adjacent Noncancerous Tissues of Non-Small-Cell Lung Cancer.

Lung tissue is directly exposed to high oxygen pressure, as well as increased endogenous and exogenous oxidative stress. Reactive oxygen species (ROS) generated in these conditions play an important role in the initiation and promotion of neoplastic growth. In response to oxidative stress, the antioxidant activity increases and minimizes ROS-induced injury in experimental systems. The aim of the present study was to evaluate the activity of antioxidant enzymes, such as superoxide dismutase (SOD; isoforms: Cu/ZnSOD and MnSOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (GR), and glutathione S-transferase (GST), along with the concentration of malondialdehyde (MDA) in tumor and adjacent noncancerous tissues of two histological types of NSCLC, i.e., adenocarcinoma and squamous cell carcinoma, collected from 53 individuals with surgically resectable NSCLC. MDA concentration was similar in tumors compared with adjacent noncancerous tissues. Tumor cells had low MnSOD activity, usually low Cu/ZnSOD activity, and almost always low catalase activity compared with those of the corresponding tumor-free lung tissues. Activities of GSH-related enzymes were significantly higher in tumor tissues, irrespective of the histological type of cancer. This pattern of antioxidant enzymes activity could possibly be the way by which tumor cells protect themselves against increased oxidative stress.

Visfatin serum concentration and hepatic mRNA expression in chronic hepatitis C.

AIM OF THE STUDY: Chronic hepatitis C (CHC) is a viral disease with metabolic disturbances involved in its pathogenesis. Adipokines may influence the inflammatory response and contribute to development of metabolic abnormalities in CHC. Visfatin exerts immunomodulatory and insulin-mimetic effects. The aim was to measure visfatin serum concentrations and its mRNA hepatic expression in non-obese CHC patients and to assess the relationships with metabolic and histological parameters. MATERIAL AND METHODS: In a group of 63 non-obese CHC patients (29 M/34 F) infected with genotype 1b aged 46.6 ±14.6 years, body mass index (BMI) 24.8 ±3.0 kg/m2, serum visfatin levels and its mRNA hepatic expression were examined and the subsequent associations with metabolic and histopathological features were assessed. RESULTS: Serum visfatin levels were significantly higher in CHC patients compared to controls (22.7 ±5.7 vs. 17.8 ±1.5 ng/ml, p < 0.001). There was no difference in serum visfatin and its mRNA hepatic expression regardless of sex, BMI, insulin sensitivity and lipids concentrations. There was no mutual correlation between serum visfatin and visfatin mRNA hepatic expression. Hepatic visfatin mRNA levels but not visfatin serum levels were higher in patients with steatosis (1.35 ±0.75 vs. 0.98 ±0.34, p = 0.009). CONCLUSIONS: Serum visfatin levels may reflect its involvement in chronic inflammatory processes accompanying HCV infection. Increased visfatin mRNA hepatic expression in patients with steatosis seems to be a compensatory mechanism enabling hepatocytes to survive metabolic abnormalities resulting from virus-related lipid droplet deposition prerequisite to HCV replication.

Clinical and prognostic value of hTERT mRNA expression in patients with non-small-cell lung cancer.

Telomerase, undetectable in normal somatic cells, plays a critical role in carcinogenesis of the majority of human tumors including lung carcinoma. The aim of our study was to determine human telomerase reverse transcriptase (hTERT) mRNA expression in patients with non-small cell lung cancer (NSCLC) in order to estimate its usefulness as diagnostic and/or prognostic factor. hTERT expression was analyzed in a group of 12 females and 28 males with NSCLC using Quantitative Real-Time Polymerase Chain Reaction (QRT-PCR method) in cancerous and non-cancerous lung tissues. Results were analyzed according to clinical data and one-, two-, and five-year survival rates. hTERT expression in the cancerous tissue was significantly higher than in the lung parenchyma free from neoplasm infiltration (p<0.05). There was no significant association between hTERT expression in the tumor tissue and age, gender, grading or clinical stage. A significant difference in hTERT expression between two types of histopathological patterns (adenocarcinoma and squamous cell carcinoma) was detected (p=0.01). No association between hTERT expression in NSCLC specimens and survival rates was found. hTERT mRNA detection by QRT-PCR in tumor and corresponding cancer-free tissues can be used as a diagnostic marker in patients with NSCLC, but seems not to be a prognostic factor.

Hepatocyte growth factor and epidermal growth factor activity during later stages of rat liver regeneration upon interferon α-2b influence.

INTRODUCTION: Liver regeneration is a complex, highly coordinated process which can be disturbed by the impact of the anti-proliferative interferon α activity. In the model of partial hepatectomy (PH) in the rat the expression of HGF and EGF genes and their molecules' tissue concentrations were analyzed in the later stages of liver regeneration (48-120 h). MATERIAL AND METHODS: 40 three-month-old male Wistar rats were randomized to groups of 20 animals each. The rats of the study group (IFN/H) were injected subcutaneously with IFNα-2b, while the control group was injected with 0.5 ml of 0.9% NaCl (NaCl/H). In the liver tissue samples obtained during hepatectomy and autopsy (regenerating liver mass) the expression of HGF and EGF genes was estimated with the Q-PCR method and the analysis of HGF and EGF molecule concentrations in tissue homogenates was conducted with the ELISA method. RESULTS: HGF but not EGF expression was significantly higher at 48 h after PH, while EGF expression was higher in normal than in regenerating liver tissue at 120 h. The analyses of correlations between expression of HGF and EGF in regenerating liver tissue, both normal and upon IFNα-2b influence, together with correlations between those factors genes' expression and HGF and EGF tissue concentrations in analyzed samples, showed no significant differences. CONCLUSIONS: HGF and EGF are not significantly involved in regulation of later stages of rat liver regeneration. IFNα-2b does not impact expression of their genes or the presence of these growth factor molecules in regenerating liver tissue.

Telomerase activity in non-small cell lung cancer.

INTRODUCTION: High telomerase activity has been detected in the majority of malignant neoplasms including lung cancer. The purpose of the study was to attempt to use telomerase activity as a prognostic factor in patients with non-small cell lung cancer (NSCLC). MATERIAL AND METHODS: Telomerase activity was analyzed in 47 tissue specimens taken from patients with NSCLC. The control group consisted of 30 specimens of non-cancerous lung parenchyma. Telomerase activity was measured by means of the telomeric repeat amplification protocol (TRAP). RESULTS: Telomerase activity in the neoplastic tissue was significantly higher than in the lung parenchyma that was free from neoplastic infiltration. There was no significant association between telomerase activity and age, gender, tobacco smoking, histological type of the tumor, or staging (pTNM). No association was found between the level of telomerase activity in NSCLC specimens and the two-year survival rate of patients (p = 0.326). A higher level of telomerase activity in poorly differentiated tumors (G3) as compared to moderately differentiated tumors (G2) was detected (p = 0.008). A positive association was identified between telomerase activity in pulmonary parenchyma free from tumor infiltration and the presence of leukocyte infiltration (p = 0.0001). CONCLUSIONS: No association was found between the level of telomerase activity in NSCLC specimens and the two-year survival rate of patients. The study has revealed a positive association between telomerase activity and the grade of differentiation (G) in NSCLC.

Omentin serum concentration and hepatic expression in chronic hepatitis C patients - together or apart?

Chronic hepatitis C (CHC) is accompanied by numerous metabolic disorders, partially associated with altered adipokine system regulation. Omentin (intelectin-1) is a novel adipokine known to play a pivotal role in metabolic regulation in CHC. In a group of 63 CHC patients (29 men/34 women) infected with genotype 1b, aged 6.6 ± 14.6 years, serum omentin levels and its gene expression in liver tissue were examined and their association with metabolic and histopathological features was assessed. Serum omentin levels were significantly higher in CHC patients compared to controls (p < 0.001), regardless of sex, body mass index (BMI), insulin sensitivity and lipid concentrations. There was no correlation between serum omentin and omentin hepatic expression. Neither parameter was associated with any histological features. Serum omentin in non-obese CHC patients seems not to be related to metabolic disorders or liver pathology. Omentin hepatic expression shows no relationship with either serum omentin levels or histopathological features. This suggests different mechanisms regulating circulating omentin concentration and omentin hepatic expression in CHC.

Prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks collected from southern Poland.

In 2008-2011 ticks were collected from southern Poland. Out of 6336 individuals collected and identified as Ixodes ricinus, 768 (2 larvae, 84 nymphs, 417 females, 265 males) were included in molecular study. The aim of this study was to investigate the prevalence and types of genospecies of Borrelia burgdorferi sensu lato in ticks. The polymerase chain reaction (PCR) was applied to detect the presence of pathogens in ticks. Subsequently the amplified DNA was digested with TasI enzyme. The infection rate was 15% (116) of examined ticks. PCR-RFLP analysis allowed distinguishing three genospecies of B. burgdorferi s.l.: B. burgdorferi sensu stricto, B. afzelii, and B. garinii. RFLP analyses of 116 positive samples revealed 96 (83%) monoinfections and 13 (11%) coinfections, whereas unidentified genospecies were present in 7 (6%) of positive samples. In the case of monoinfections, B. burgdorferi s.s. was the predominant species of pathogen in infected ticks - 61.4%. Other genospecies: B. garinii and B. afzelii were detected in 22.9% and 15.6% of the samples, respectively. To sum up, 15 % of ticks were infected by B. burgdorferi s.l which increases the risk of human infections in the recreational areas of southern Poland. Furthermore, there is a need to increase public awareness and implement more preventive measures concerning Lyme disease.

Hepatic chemerin and chemokine-like receptor 1 expression in patients with chronic hepatitis C.

INTRODUCTION: Chemerin seems to be involved in pathogenesis of chronic hepatitis C (CHC). Hepatic expressions of chemerin and its receptor, chemokine receptor-like 1 (CMKLR1), in CHC have not been studied so far. AIM: To evaluate chemerin and CMKLR1 hepatic expression together with serum chemerin concentration in CHC patients and to assess their relationship with metabolic and histopathological abnormalities. METHODS: The study included 63 nonobese CHC patients. Transcription of chemerin and CMKLR1 was assessed by quantitative real-time PCR, while serum chemerin was assessed by enzyme-linked immunosorbent assay. RESULTS: Expression of chemerin and CMKLR1 was present in the liver of all CHC patients regardless of sex or age. This expression was not associated with necroinflammatory activity and steatosis grade, fibrosis stage, and metabolic abnormalities. There was a negative association between serum chemerin and chemerin hepatic expression (r = (-0.41), P = 0.006). CONCLUSION: The study for the first time confirmed a marked expression of chemerin and CMKLR1 in the liver of CHC patients. The study was performed using the homogenates of human liver tissue, so it is not possible to define whether hepatocytes or other cell types which are abundantly represented in the liver constitute the main source of chemerin and CMKLR1 mRNA.

The activity of antioxidant enzymes in colorectal adenocarcinoma and corresponding normal mucosa.

Oxidative stress is one of several factors which contribute to the development of colorectal carcinogenesis. The aim of the study was an assessment of the activity of antioxidant enzymes in tumour and corresponding normal distal mucosa in a group of patients with colorectal adenocarcinoma. Samples of tumour and corresponding normal mucosa were obtained during a resection of colorectal cancer from 47 patients aged between 26 and 82 years. The average distance of corresponding normal distal mucosa from the tumour was 4.49 cm. Activities of antioxidant enzymes: superoxide dismutase (SOD), glutathione reductase (GR), glutathione peroxidase (GPx), glutathione S-transferase (GST), and catalase (CAT) were measured in tissue homogenates. The patients were grouped according to the tumour stage (Duke's staging), grading, localization, and size of tumour, as well as age and sex. Statistical analysis was performed. The activity of SOD and GPx was considerably increased, while the activity of GST decreased significantly in tumour as compared with normal mucosa. GR activity in colorectal cancer was evidently higher in tumours of proximal location compared with the distal ones. The distance of corresponding normal distal mucosa from the tumour was analyzed and related to all assayed parameters. A decreased GST activity was observed in corresponding normal mucosa more than 5 cm distant from the tumour in patients with CD Duke's stage. The higher activity of superoxide dismutase and glutathione peroxidase in tumour compared to corresponding normal mucosa could indicate higher oxidative stress in colorectal adenocarcinoma cells.

[The concentration of HSP 27 in tumour and adjacent distal mucosa in colorectal adenocarcinoma]. / Stezenie bialka HSP 27 w guzie oraz marginesie dystalnym w gruczolakoraku jelita grubego.

OBJECTIVE: Heat shock proteins (HSPs) play a very important role in eukaryotic and prokaryotic cells. They are produced in cells constitutively or induced under stress conditions. During carcinogenesis, HSPs have been reported to show alteration of their expression levels. AIM OF THE STUDY: The aim of the study was assessment of the concentration of HSP 27 in tumour and adjacent distal normal mucosa in colorectal adenocarcinomas. MATERIAL AND METHODS: Samples of tumour tissues and adjacent distal normal colon tissues were obtained during resection operations from 47 patients aged between 26-82. Average distance of adjacent distal normal mucosa from the tumour was 4.49 cm. In tissues homogenate total protein level and concentration of HSP 27 were assayed. The patients were grouped according to clinical classification (Duke's staging), grade of differentiation, localization, and size of tumour, as well as age and sex. Relation between adjacent distal normal mucosa collected in different distances from the tumour and compared to concentration of HSP 27 was analyzed. Statistical analysis was performed. RESULTS: The concentration of HSP 27 was significantly increased in adjacent distal normal tissues compared with tumour tissues. Moreover, the concentration of HSP 27 was increased in adjacent normal tissues in patients with CD Duke's staging compared to patients with AB Duke's staging. No differences in concentration of HSP 27 between tumour and normal tissues compared with grade of differentiation, localization, and size oftumour, as well as age and sex were observed. No relation between adjacent distal normal mucosa collected in different distances from the tumour and compared to concentration of HSP 27 was observed. CONCLUSIONS: The higher concentration of HSP 27 in adjacent distal normal mucosa can be a response to stress factors related with metabolic changes of cancer tissues or can be a protective function to stress factors of the tumour.

TNF-alpha expression in gastric mucosa of individuals infected with different virulent Helicobacter pylori strains.

BACKGROUND: Helicobacter pylori colonizes the human gastric mucosa, leading to chronic superficial gastritis and in some cases to peptic ulceration, gastric adenocarcinoma, or gastric lymphoma. It has been postulated that the clinical outcome depends on differences in H. pylori strain virulence as well as on individual factors of the host. Thus, we aimed to assess the relation between H. pylori cagA/vacA genotypes and the TNF-alpha gene expression in gastric mucosa specimens from patients with chronic gastritis. MATERIAL/METHODS: This study was conducted with gastric mucosa samples obtained during gastroendoscopy from 43 H. pylori-infected individuals with chronic gastritis. The presence of ureA and cagA genes and vacA allele combinations were analyzed in isolated DNA by the polymerase chain reaction method. Isolates of RNA were used for cDNA synthesis by reverse transcription. Synthesized cDNA was used to determine the TNF-alpha gene expression level by quantitative real-time polymerase chain reaction assay. RESULTS: The cagA gene was detected in 67.44% of H. pylori strains. Five vacA alleles in the tested H. pylori strains were detected: s1a/m1 (18.60%), s1a/m2 (23.26%), s1a/m3 (18.60%), s1b/m2 (6.98%), and s2m2 (32.56%). There were no significant differences in TNF-alpha gene expression in strains expressing cagA versus those not expressing this gene, and no significant differences among strains with different vacA alleles. CONCLUSIONS: TNF-alpha gene expression in the gastric mucosa of H. pylori-infected patients appears to be independent of H. pylori cagA/vacA genotype.

Methylation of the MGMT and p16 genes in sporadic colorectal carcinoma and corresponding normal colonic mucosa.

BACKGROUND: Colorectal cancer, one of the most aggressive cancers, occurs with a high incidence in most countries. The usual treatment is surgery and subsequent chemotherapy and radiotherapy. Cancer development and progression is dictated by series of alterations in genes such as tumor suppressor genes, DNA repair genes, oncogenes and others. In colorectal carcinogenesis disturbances different from mutations called an epigenetic regulation are also taken into consideration. Epigenetics is defined as a modifications of the genome, heritable during cell division, which do not involve a change in the DNA sequence. In our study we analyzed methylation of CpG islands in the MGMT and p16 genes in sporadic colorectal cancers and normal corresponding colonic mucosa. MATERIAL/METHODS: Fresh tissue samples were obtained from 68 patients (age of 23 to 81 years) with primary colorectal adenocarcinoma and corresponding normal tissues. We used methylation-specific polymerase chain reaction (MSP) for analysis of the methylation status of MGMT and p16. RESULTS: Methylation of MGMT and p16 was detected in 59% and 53% of tumors, respectively. In corresponding normal colonic mucosa methylation of MGMT was detected in 20% and p16 in 18%. The normal colon mucosa obtained from younger patients (age of <65 years) showed less methylation frequency as compared with the normal mucosa from the older ones (age of >65 years). CONCLUSIONS: The older age and female gender are generally associated with higher methylation levels for most CpG islands in normal colonic mucosa. These results indicate that MGMT and/or p16 aberrant methylation may play an important role in colorectal cancer.

Cellular toxicity of oxycholesterols.

Oxycholesterols (OS) are formed from cholesterol or its immediate precursors by enzymatic or free radical action in vivo, or they may be derived from food. OS exhibit a wide spectrum of biological activities. In OS cytotoxicity, several mechanisms seem to be involved: e.g. inhibition of HMG-CoA reductase activity, antiproliferative action, apoptosis induction, replacement of cholesterol by OS in membranes followed by changes in cellular membrane structure and functionality, and immune system functions alteration. Furthermore, OS may be mutagenic and carcinogenic and may serve as intracellular signaling or regulatory molecules. Here we review OS cellular activities with special attention to the cytotoxic action in vivo and in vitro using experimental models.

[Presence of spirochetes of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks in the recreational area of Tarnowskie Gory and Zabrze districts in 2001-2003]. / Obecnosc kretków Borrelia burgdorferi sensu lato u kleszczy Ixodes ricinus na terenach rekreacyjnych okolic Tarnowskich Gor i Zabrzaw latach 2001-2003.

Borrelia burgdorferi sensu lato is an aethiological factor of borreliosis. Borrelia spirochete is transmitted to humans by ticks. The aim of the study was to estimate the spirochetes B. burgdorferi s.1. infection in Ixodes ricinus ticks by using PCR analysis of the flagellin gene fragment. The study was carried out in 2001-2003 in recreational areas of the Silesian Region in Poland (Tarnowskie G6ry and Zabrze administrative districts). In the 233 ticks analysed the DNA of B. burgdorferi s.l. was detected in 33 (14,2%) cases. Further comparison of spirochetes infection percentages in ticks during the three consecutive vegetation seasons was made. The investigation of B. burgdorferi s.l. detection in tick populations of recreational and endemic areas is important for the evaluation of the risk of infection.

[Molecular analysis of methicillin-resistant staphylococci isolated from hospital patients and in the out-patient clinic]. / Molekularna analiza metycylinoopornych gronkowców izolowanych od chorych hospitalizowanych i ambulatoryjnych.

In this study, a molecular analysis of the methicillin-resistant coagulase-negative staphylococci strains was performed. The obtained results of the biochemical and drug resistant pattern investigations were insufficient to assess the relationship between the strains. Therefore genotyping by the restriction fragment length polymorphism analysis (PCR-RFLP) method was performed. Analyzed strains characterized presence of the mecA gene-PCR products. The PCR products were digested with DraI and TasI, and the fragments separated by agarose gel electrophoresis. Typing of the methicillin-resistant gene using PCR-RFLP showed that all MRCNS strains possess an identical restriction pattern of the mecA gene. This identical restriction pattern of the mecA gene in investigated strains may suggest an easy transfer of this gene between different staphylococci species and lead to the spreading of methicillin-resistant among hospital strains. Furthermore performing the comparison of different phenotype and genotype methods has shown that the PCR-RFLP method is quick and reliable, enabling the detection and estimation of the relationship between MRCNS strains.

[Prevalence of Borrelia burgdorferi sensu lato in ticks, Ixodes ricinus in Tarnowskie Góry district]. / Czekstosc wystepowania Borrelia burgdorferi sensu lato u kleszczy Ixodes ricinus z okolic Tarnowskich Gór.

Borrelia burgdorferi is an aetiological factor of borreliosis (Lyme disease). The main vectors of Borrelia burgdorferi are larvae, nymphs, and females of Ixodes ricinus. The aim of this paper was to analyse infection parameters of Borrelia burgdorferi in a selected populations of Ixodes ricinus. The study was conducted in Tarnowskie Góry administrative district (Krupski Mlyn, Zielona, Lubliniec, Tarnowskie Góry, Swierklaniec, Tworóg, and Zbroslawice). A total of 85 ticks were collected with a piece of cloth dragged over the vegetation. The Borrelia burgdorferi infection was confirmed with a PCR method, using flagellin protein gene DNA amplification. DNA of Borrelia burgderferi sensu lato was present in 14 ticks, which constituted 16.5% of the population studied. The percentage of infected females and nymphs was 26.8%, 22.2%, and 5.6%, respectively. A high prevalence of the pathogen (50%) was in ticks revealed in the recreation areas and the community forest in Swierklaniec.