Circulating matrix metaloproteinases as biomarkers in colorectalcancer.

The aim of this research is to offer comprehensive point of view related to perspective tumor markers called matrix metaloproteinases and their natural tissue inhibitors. Those markers are potentially useable mainly in postoperative follow-up in patients with colorectal cancer.

Can the One-Step Nucleic Acid Amplification method of lymph nodes examination make the staging of pulmonary tumours more precise?

INTRODUCTION: The aim of this article is to compare the sensitivity of detecting micrometastases in hilar and mediastinal lymph nodes in case of primary (non-small cell) and secondary (metastases of colorectal carcinoma) pulmonary tumours using standard histopathological examination with haematoxylin-eosin staining, immunohistochemistry examination with Anti-Cytokeratin 19 antibody and examination based on the One-Step Nucleic Acid Amplification method. METHOD: During radical surgical treatment of primary non-small cell lung carcinoma and pulmonary metastases of colorectal carcinoma, hilar and mediastinal lymph nodes of 100 patients enrolled in the study in the period from 2015 to 2017 were extracted based on a standard classification. These lymph nodes were subsequently divided along the longitudinal axis into 4 identical parts where part one and three on the left were intended for examination based on the One-Step Nucleic Acid Amplification method, whereas parts two and four were subjected to histopathological examination. In evaluating the respective parts of the nodes by histological examination, the nodes were first examined by a standard procedure that involves haematoxylin-eosin staining, followed by immunohistochemistry examination with Anti-Cytokeratin 19 antibody. The One-Step Nucleic Acid Amplification method was performed in the kit supplied by Sysmex (Kobe, Japan) and is based on the detection of cytokeratin 19 mRNA (messenger ribonucleic acid) by reverse transcription coupled with isothermal amplification. RESULTS: A total of 1,426 lymph nodes of the patients enrolled in the study were extracted and examined using the above mentioned methodology. In 78 patients (78%), identical results were obtained using haematoxylin-eosin staining, immunohistochemistry with Anti-Cytokeratin 19 and One-Step Nucleic Acid Amplification. Micrometastases in the lymph nodes using the One-Step Nucleic Acid Amplification method in the absence of the other methods were proven in 16 patients (16%). Only in 3 cases (3%), the examination by haematoxylin-eosin staining, or immunohistochemistry with Anti-Cytokeratin 19, was positive while One-Step Nucleic Acid Amplification was negative. The results obtained by immunohistochemistry with Anti-Cytokeratin 19 antibody were practically the same as those obtained by haematoxylin-eosin staining (97%). CONCLUSION: The results of the study have demonstrated a higher percentage of metastases detected in hilar and mediastinal lymph nodes if the One-Step Nucleic Acid Amplification method of examination was used compared to haematoxylin-eosin staining and immunohistochemistry with Anti-Cytokeratin 19 antibody (upstaging in 16%). This shows that the examination of lymph nodes using the One-Step Nucleic Acid Amplification method can have a certain potential to make the pulmonary tumours staging more accurate. On the other hand, immunohistochemistry with Anti-Cytokeratin 19 antibody seems to be not so useful. However, it is necessary to prove this hypothesis in follow-up studies, or where applicable, in a larger cohort of patients. Another task is to ascertain, by careful patient monitoring, the influence of the micrometastases detected in their lymph nodes using the One-Step Nucleic Acid Amplification method on these patients' follow-up. Key words: lung cancer - lymph nodes - H&E - IHC CK19 - OSNA assay.

[One-Step Nucleic Acid Amplification method - what is the future of sentinel lymph node management?] / Metoda One-Step Nucleic Acid Amplification - jaká je budoucnost zpracování sentinelových uzlin?

OBJECTIVE: The article summarizes current possibilities of usage of the One-Step Nucleic Acid Amplification method (OSNA) in the perioperative management of sentinel lymph nodes in oncologic surgery. The principle of this method is the detection of cytokeratin 19 (CK19) in the lymphatic tissue as a marker of the metastatic spread. DESIGN: Review article. SETTINGS: Department of Obstetrics and Gynaecology, University Hospital Pilsen, Faculty of Medicine in Pilsen, Charles University, Prague; Department of Biology, Faculty of Medicine in Pilsen, Charles University, Prague; Department of Immunochemistry, University Hospital Pilsen, Faculty of Medicine in Pilsen, Charles University, Prague; Sikl´s Department of Pathology, University Hospital Pilsen, Faculty of Medicine in Pilsen, Charles University, Prague. METHODS: The review of the literature published until the end of April 2017 available on the PubMed database was performed. The official abbreviation OSNA and the full name of the method One-Step Nucleic Acid Amplification was used for search in this database. CONCLUSION: The usage of the OSNA method with the detection of CK 19 in the sentinel lymph nodes as a marker of metastatic spread to the lymphatic tissue currently represents an acceptable form of perioperative sentinel lymph node management in patients with breast and colorectal cancer. Until now published data are pointing towards possible successful application of this method in sentinel lymph node management in patients with some other malignancies, such as thyroid carcinoma, gastric cancer, uterus cancer and head and neck cancer. More data is needed to establish this method also in those neoplasms.

Prognostic role of serum C-reactive protein in patients with advanced-stage NSCLC treated with pemetrexed.

Pemetrexed is an intravenously administered antifolate cytostatic agent targeting several folate-dependent enzymatic pathways, widely used in the treatment of patients with advanced non-small cell lung cancer (NSCLC). It has been previously demonstrated that the superiority of pemetrexed is limited to patients with non-squamous histology. Aside from the non-squamous histology, there is still no available molecular biomarker predicting treatment efficacy of pemetrexed-based chemotherapy. The aim of our retrospective study was to evaluate the association of baseline serum levels of C-reactive protein (CRP) with outcomes in a large cohort of patients with non-squamous NSCLC treated with pemetrexed. Clinical data of 325 patients were analysed. Serum samples were collected within one week before the initiation of treatment. The median progression-free (PFS) and overall survival (OS) for patients with high CRP was 2.1 and 9.5 compared to 4.2 and 20.5 months for those with normal CRP (p=0.002 and p<0.001, respectively). The multivariable Cox proportional hazards model revealed that serum CRP (HR=1.46, p=0.002) was significantly associated with PFS and also with OS (HR=1.95, p<0.001). In conclusion, the study results suggest that pretreatment serum CRP is associated with poor outcome of non-squamous NSCLC patients treated with pemetrexed.

Serum albumin is a strong predictor of survival in patients with advanced-stage non-small cell lung cancer treated with erlotinib.

Molecular targeted therapy based on tyrosine kinase inhibitors (TKI), directed at epidermal growth factor receptor (EGFR) is one of the novel effective agents in management of advanced-stage of Non Small Cell Lung cancer (NSCLC). However several candidate predictors have been extensively studied, apart from activating EGFR gene mutations, no reliable biochemical or molecular predictors of response to erlotinib have been validated. The aim of our retrospective study was to evaluate the association of baseline serum albumin with outcomes in a large cohort of patients with advanced-stage NSCLC treated with erlotinib. Clinical data of 457 patients with locally-advanced (III B) or metastatic stage (IV) NSCLC treated with erlotinib were analysed. Serum samples were collected and the measurement was performed one day before the initiation of erlotinib treatment. Before the treatment initiation, low albumin was (<35 g/l) measured in 37 (8.1%) patients and normal albumin (≥ 35 g/l) was measured in 420 (91.9%). The median PFS and OS for patients with low serum albumin was 0.9 and 1.9 months compared to 1.9 and 11.4 months for patients with normal serum albumin (p=0.001 and p<0.001). The multivariate Cox proportional hazards model revealed that EGFR mutation status (HR=2.50; CI: 1.59-3.92; p<0.001) and pretreatment serum albumin (HR=1.73; CI: 1.21-2.47; p=0.003) were significant independent predictive factors for PFS, whereas EGFR mutation status (HR=3.14; CI: 1.70-5.81; p<0.001), stage (HR=1.48; CI: 1.09-2.02; p=0.013), ECOG PS (HR=1.77; CI: 1.37-2.29; p<0.001) and pretreatment serum albumin (HR=4.60; CI: 2.98-7.10; p<0.001) were significant independent predictive factors for OS. In conclusion, the results of present retrospective study indicate that pretreatment hypoalbuminemia is associated with poor outcome of NSCLC patients treated with erlotinib. Based on these results, measuement of serum albumin is an objective laboratory method feasible for estimation of prognosis of patients with advanced-stage NSCLC.

Desphospho-uncarboxylated matrix Gla protein is associated with increased aortic stiffness in a general population.

Matrix Gla protein (MGP), a natural inhibitor of calcification, strongly correlates with the extent of coronary calcification. Vitamin K is the essential cofactor for the activation of MGP. The nonphosphorylated-uncarboxylated isoform of MGP (dp-ucMGP) reflects the status of this vitamin. We investigated whether there is an association between dp-ucMGP and stiffness of elastic and muscular-type large arteries in a random sample from the general population. In a cross-sectional design, we analyzed 1087 subjects from the Czech post-MONICA study. Aortic and femoro-popliteal pulse wave velocities (PWVs) were measured using a Sphygmocor device. Dp-ucMGP concentrations were assessed in freshly frozen samples by enzyme-linked immunosorbent assay methods using the InaKtif MGP iSYS pre-commercial kit developed by IDS and VitaK. Aortic PWV significantly (P<0.0001) increased across the dp-ucMGP quartiles. After adjustment for all potential confounders, aortic PWV independently correlated with dp-ucMGP (with beta coefficient (s.d.) 11.61 (5.38) and P-value=0.031). In a categorized manner, subjects in the top quartile of dp-ucMGP (⩾ 671 pmol l(-1)) had a higher risk of elevated aortic PWV, with corresponding adjusted odds ratio (95% confidence interval) 1.73 (1.17-2.5). In contrast, no relation between dp-ucMGP and femoro-popliteal PWV was found. In conclusion, increased dp-ucMGP, which is a circulating biomarker of vitamin K status and vascular calcification, is independently associated with aortic stiffness, but not with stiffness of distal muscular-type arteries.

Isocitrate dehydrogenase-1 mutations as prognostic biomarker in glioblastoma multiforme patients in West Bohemia.

INTRODUCTION: Glioblastoma multiforme (GBM) is the most malignant primary brain tumor in adults. Recent whole-genome studies revealed novel GBM prognostic biomarkers such as mutations in metabolic enzyme IDH-isocitrate dehydrogenases (IDH1 and IDH2). The distinctive mutation IDH1 R132H was uncovered to be a strong prognostic biomarker for glioma patients. We investigated the prognostic role of IDH1 R132H mutation in GBM patients in West Bohemia. METHODS: The IDH1 R132H mutation was assessed by the RT-PCR in the tumor samples from 45 GBM patients treated in the Faculty Hospital in Pilsen and was correlated with the progression free and overall survival. RESULTS: The IDH1 R132H mutation was identified in 20 from 44 GBM tumor samples (45.4%). The majority of mutated tumors were secondary GBMs (16 in 18, 89.9%). Low frequency of IDH1 mutations was observed in primary GBMs (4 in 26, 15.3%). Patients with IDH R132H mutation had longer PFS, 136 versus 51 days (P < 0.021, Wilcoxon), and OS, 270 versus 130 days (P < 0.024, Wilcoxon test). SUMMARY: The prognostic value of IDH1 R132H mutation in GBM patients was verified. Patients with mutation had significantly longer PFS and OS than patients with wild-type IDH1 and suffered more likely from secondary GBMs.

[Determination of anti-Müllerian hormone in women]. / Stanovení anti-Müllerova hormonu u zen.

Anti-Müllerian hormone (AMH) is a glycoprotein and belongs to the TGF-ß growth factors family. Our review describes the method of AMH determination in serum and follicular fluid. The reference values and changes in AMH levels during a womans life are also discussed. In addition, it is also presented the relationship between AMH, obesity, smoking and use of hormonal contraceptives. The focus of the work is the importance of the determination of AMH in clinical practice. In assisted reproduction has become its determination one of the tools to detect ovarian reserve. It helps not only predict reduced response to stimulation with gonadotropins but also the risk of the ovarian hyperstimulation syndrome. Benefits of the ovarian reserve detection using AMH serum levels are discussed in comparison with the antral follicle count (AFC) determined by ultrasound. Several clinical indications of AMH determination are mentioned in the next section. These are primarily the polycystic ovary syndrome (PCOS), which is a great challenge not only for the AMH testing, but there is an open space for further interdisciplinary cooperation. Endometriosis has no direct effect on ovarian reserve and AMH levels in serum. AMH is very sensitive tumor marker in the diagnostics and monitoring of ovarian granulosa cells tumors. Treatment of cancer disease burdens entire body, including healthy cells. Ovarian follicles are very sensitive to chemotherapy and radiation. AMH is a good predictor of ovarian reserve damage during radio- and chemotherapy.

[HE4 a biomarker of ovarian cancer]. / HE4 - biomarker ovariálního karcinomu.

OBJECTIVE: Verification of the importance of determination of HE4 and calculation of ROMA index for increasing the efficiency of diagnosis of ovarian cancer in a population of Czech women. DESIGN: Prospective study. SETTING: Department of Gynaecology and Obstetrics, Faculty Hospital in Pilsen. METHODS: In the period from 06/24/2010 to 12/01/2011 was at the Department of Gynaecology and Obstetrics, University Hospital Pilsen examined 552 patients with abnormalities in the pelvis. Patients were divided into two groups. There were 30 women with histologically confirmed malignant ovarian tumors. Another 522 women had benign findings. According to the levels of FSH were women in both groups divided into premenopausal and postmenopausal. At all women were measured CA 125, HE4 and FSH. HE4 and CA125 were determined using the chemiluminescent device Architect 1000 (Abbott, USA), FSH chemiluminescent method on the device DXI 800 (Beckman Coulter, USA). At all premenopausal women was calculated ROMA1 index and at all postmenopausal women ROMA2 index. SAS statistical software 9.2 were used for all statistical calculations. RESULTS: The highest diagnostic efficiency was achieved by a combination of HE4 and CA125 markers with the calculation ROMA2 index for postmenopausal women. In determining of menopausal status according to the values of FSH cut-off for menopause 40 IU/L and cut-off at 26.4% for ROMA2 reaches ROMA2 sensitivity of 92.3%, specificity of 88.5% and PV- of 99.3%. If we reduce the cut-off for laboratory diagnosis of menopause using FSH at 22 IU/L, and cut-off for ROMA2 was 26.3% reaches ROMA2 sensitivity of 95.2%, specificity of 87.8% and PV- of 99.5%. CONCLUSION: HE4 in combination with CA125 and current ROMA index calculation is a suitable methodology to improve the detection of ovarian cancer.

[Vitamin D and neurological diseases]. / Vitamin D a neurologická onemocnení

We provide an overview of the association between vitamin D and some neurological diseases where the correlation has repeatedly been described. The majority of literature refers to cerebrovascular diseases, followed by multiple sclerosis and cognitive disorders. Vitamin D hypovitaminosis might be associated with the diseases directly or it might contribute to the disease risk factors (typically in cerebrovascular events). Vitamin D hypovitaminosis may also play a role in patients with residual functional involvement due to a neurological disorder (movement disorders, lack of self-sufficiency) and worsen functional status owing to muscle weakness, instability and falls.

[Will vitamin D become a new antidiabetic?]. / Stane se vitamin D novým antidiabetikem?

Based on experience from experimental and human studies, vitamin D can be considered an important factor lowering the risk of diabetes mellitus type 1 and 2. The mechanism consists in the direct influence of vitamin D via nuclear receptors on genes coding proteins associated with normal function of B cells of Langerhans islands and genes coding proteins ensuring normal function of the immune system. There is also an indirect influence via regulation of homeostasis of calcium. Clinical observation and cross-sectional studies show an inverse relationship between vitamin D deficiency and appearance of diabetes mellitus type 2. A major deficit of vitamin D in diabetes mellitus type 2 appears to be an independent factor able to predict an increased risk of future cardiovascular mortality. Deficiency in early periods of life was shown to precede autoimmune diabetes mellitus type 1 in an experiment as well as in humans. Prevention of vitamin D deficiency in early as well as later periods of life is a basic pre-requisites of successful preventive measures against diabetes mellitus type 1. Explicit evidence for the significance of the correction of vitamin D deficiency for improvement of metabolic control in diabetics is still missing mainly due to a low number of intervention, placebo-controlled and randomized trials. On the other side, intervention studies often showed positive influence on the conditions accompanying diabetes, such as systolic hypertension or endothelial dysfunction. Unlike in diabetics, the intervention trials showed positive results in non-diabetics with high risk of type 2 diabetes and impaired fasting glycaemia or insulin resistance. One can conclude that existing knowledge already indicate that maintaining the level of 25-hydroxyvitamin D > 30 ng/ml during the year without seasonal variations will be have multiple real as well as potential health benefits. A great promise for clinical practice are the structural analogs of vitamin D tested experimentally, which maintain the influence on the immune system, effect of insulin and B cells function, but have suppressed influence on bone and calcium metabolism.

Cytokeratin serum biomarkers in patients with colorectal cancer.

BACKGROUND: Circulating cytokeratins have shown to be important for management of patients with lung cancer. Here we investigated their role for differential diagnosis, therapy monitoring and prognosis in colorectal cancer (CRC). PATIENTS AND METHODS: Pretherapeutic levels of cytokeratin-19 fragments (CYFRA 21-1), carcino-embryonic antigen (CEA) and cancer antigen (CA) 19-9 were measured in 42 patients with CRC, 45 with benign colorectal diseases and 51 healthy controls. Furthermore, courses of CYFRA 21-1, tissue polypeptide antigen (TPA), tissue polypeptide specific antigen (TPS), M30-antigen, CEA and CA 19-9 were analyzed in prospectively collected sera of 15 patients with CRC during primary chemotherapy and were correlated with therapy response and overall survival (OS). RESULTS: Similar to CEA and CA 19-9, CYFRA 21-1 was significantly elevated in serum from patients with CRC (median 2.1 ng/ml) as compared with healthy (1.2 ng/ml; p<0.0001) and benign gastrointestinal controls (1.7 ng/ml; p=0.0178) and showed stage dependency in CRC (p=0.0118). CYFRA 21-1 correlated with CEA in benign diseases and CRC but not with CA 19-9. The best discrimination between healthy controls and patients with CRC was achieved by combination of CYFRA 21-1 and CA 19-9 (area under the curve; AUC=86.7%), while the combination of CEA and CA 19-9 discriminated best between benign diseases and CRC (AUC=73.9%). In CRC patients during primary chemotherapy, levels of cytokeratins CYFRA 21-1, TPA, TPS, CEA and CA 19-9 tended to be higher in patients with poor response to therapy and with poor prognosis. CONCLUSION: Cytokeratins are elevated in patients with CRC and show some association with response to primary therapy and prognosis.

Matrix metalloproteinases and their inhibitors in correlation to proliferative and classical tumour markers during surgical therapy of colorectal liver metastases.

OBJECTIVES: Classical and proliferative tumour markers and matrix metalloproteinases and their tissue inhibitors reflect the features of malignancy and are useful in prediction of prognosis in patients with colorectal liver metastases. There is very limited information about their physiological functions during regeneration and healing of liver parenchyma after any type of liver surgery for malignancy. METHODS: The presented study included the patients, who underwent following surgical procedures for CLM, benign liver lesions and inguinal hernias: Group A: 22 patients with inguinal hernias, Group B: 26 patients with benign liver lesions, Group C: 30 patients with colorectal liver metastases (CLM) who were treated by radiofrequency ablation, Group D: 41 patients with CLM who underwent a radical surgical therapy - resection, and Group E: 22 patients with inoperable CLM who underwent an explorative laparotomy without any surgical procedure. RESULTS: The preoperative and postoperative serum levels of CEA, CA 19-9, TK, TPA, TPS, MMP-2, MMP-9, TIMP-1, and TIMP-2 were statistically analyzed and compared within the groups to estimate the influence of a surgical procedure type. These results reflect the influence of surgical procedure on the serum levels of studied tumour markers during operation. CONCLUSIONS: It was the first description using these types of comparison to all metalloproteinases, their inhibitors, and proliferative and classical tumour markers. It could help us to estimate the critical relations of these tumour markers in prognoses of disease free survival or overall survival in patients after a surgical procedure for CLM (Tab. 5, Ref. 26).

[Can tumor markers predict outcomes of portal vein branch embolization in patients with primary inoperable liver tumors?]. / Mohou nádorové markery predpovídat výsledek embolizace vetve portální zíly u nemocných s primárne inoperabilním nádorem jater?

INTRODUCTION: Portal vein embolization (PVE) is one of the options to increase the number of resecable cases in patients with primary inoperable liver tumors. However, insufficient growth of liver parenchyma or postoperative tumor progression remains problematic in PVE procedures. Generally, tumor markers are of significance in patient postoperative monitoring for the disease recurrence. The aim of this study is to assess the potential of tumor markers in predicting PVE outcomes. METHOD: The study group included 43 subjects with primary or secondary tumors, in whom serum alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), thymidine kinase (TK), tissue polypetide antigen (TPS) and MonoTotal levels were assessed 28 days following PVE. The liver parenchyma growth or tumor progression were assessed based on computer tomography. RESULTS: Sufficient liver parenchyma hypertrophy was recorded in 27 (62.8 %) patients with subsequent liver resection. Insufficient post-PVE liver parenchyma growth was recorded in 5 (11.6 %) patients and tumor progression was recorded in 11 (25.6 %) subjects. The following tests were considered significant predictive tumor markers of PVE outcomes: serum levels of CEA, TPA, Mono Total prior to PVE, and serum levels of TK, TPA, Mono Total within 28 days following PVE. CONCLUSION: Tumor markers may be significant in predicting PVE outcomes in patients with primary inoperable liver tumors. However, in order to make final conclusions on their clinical significance, larger patient group studies should be performed.

[Plasmatic levels of proinflammatory cytokines in abdominal aortic aneurysms]. / Plazmatické hladiny prozánetlivých cytokinu u aneuryzmatu abdominální aorty.

INTRODUCTION: Inflammation within the abdominal aortic wall is generally considered a very significant ethiopathogenic factor in the development of abdominal aortic aneurysms. Proinflammatory cytokines are important mediators of inflammation within the abdominal aortic wall. AIM: The aim of the study was to research, whether plasmatic levels of certain proinflammatory cytokines, which can commonly be evaluated (TNFalpha, IL-1, -2, -6 a -8), play a significant role in the development of AAA. METHOD: The prospective non-randomized study included 345 patients with AAAs. The patients were assigned to 5 subgroups based on their symptoms and AAA diameters. The first subgroup included patients with symptomatic AAAs, including AAA ruptures (N = 69), the second subgroup included subjects with asymptomatic AAAs (N = 276) with AAA diameters up to 5 cm (N = 72), the third subgroup included 5 cm (N = 72), the fourth included 5-8 cm (N = 192) and the fifth subgroup included subjects with AAA diameters of more than 8 cm (N = 81). The mean age of patients was 74.1 +/- 7.8 years (56-84 y.o.a.). The male to female ratio was 5:1. The control group included 30 healthy volunteer subjects of similar age and male to female rates, who had no clinical signs of arterial disorders. Plasmatic levels of cytokines were evaluated from venous blood samples using ELISA (Bender, Austria) testing. Statistical assessment of the results was performed using ANOVA and Wilcoxon tests with Spearman's correlation. P values < 0.05 were considered significant. RESULTS: Plasmatic concentrations of proinflammatory cytokines were found to be statistically significantly higher in patients with AAAs compared to those in healthy volunteers. Plasmatic IL8 levels were significantly decreasing proportionally to decreasing AAA diameters (p < 0.05). TNFalpha levels were found to be significantly low in symptomatic patients with AAA ruptures (p < 0.05). CONCLUSION: The study confirmed the significance of proinflammatory cytokines levels monitoring in AAA patients. The authors showed that, for instance IL8 activity and to a certain extent TNFalpha activity, is the highest in small and developing AAAs. These findings would be significant for customized medication therapy aimed at blocking the effects of these factors on the inflammatory process within the AAA wall.

Signaling molecules for early detection of adverse interactions during mechanical ventilation in animal models.

AIM: The early identification of adverse interactions during mechanical ventilation, investigated by multiplexed immunoanalysis. MATERIALS AND METHODS: Twenty piglets (average age 7 weeks, weight 23 kg) were intubated and divided into groups: A, spontaneously breathing; B, protectively ventilated; C, ventilated with injurious strategy; D, ventilated with lung disability. At the 1st hour (time-1) and 12th hour (time-2) of the study, brain natriuretic peptide (BNP), intercellular cell adhesion molecules (ICAM-1), vascular cell adhesion molecules (VCAM-1), tumor necrosis factor-alpha (TNF-alpha), and interleukin-6 (Il-6) were analyzed in the blood. RESULTS: The injurious ventilated group C exhibited an increase in both cell adhesion molecules (p<0.01), TNF-alpha and BNP (p<0.05) at time-1, and at time-2 further increases (p<0.05). In group D, an increase in ICAM-1 and BNP (p<0.05) at time-1, and increases in Il-6 and ICAM-1 (p<0.05) at time-2, with notable decreases in urine output were observed. Overall, the lung damage correlated with TNF-alpha (r=0.904), Il-6 (r=0.740), and ICAM-1 (r=0.756) levels. CONCLUSION: All five monitored molecules quickly and reliably signaled adverse interactions.

[IGF1 and tumor markers in different breast cancer stages]. / IGF1 a nádorové markery ve stadiích nádoru prsu.

INTRODUCTION: In our work we asked ourselves whether it would be possible to use growth factors for a quick orientation in the clinical status of patients prior to biopsy and histological examination. MATERIAL AND METHODS: Our patient group included 82 patients with breast cancer. Serum samples were collected preoperatively. Histological examination findings were available for each patient. Our set was divided into three groups based on the disease stage. The values of analytes in different tumor stages were statistically evaluated and statistical comparisons of Stage I and II, and then of Stage II and III were performed. RESULTS: Tumor markers CEA, CA 15-3, TK, TPA-M and MonoTotal correlate with the disease severity. Serum levels of the growth factor IGFI negatively correlated with the severity of cancer. There was aa statistically significant increase in the EGF growth factor serum levels between Stage I and II. No statistically significant differences between Stage I vs. II and Stage II vs. III were detected when HGF and VEGF growth factor serum levels were assessed. CONCLUSION: The growth factor EGF is one of the candidates to become a tumor growth marker in early disease stages. The IGFI, HGF and VEGF growth factors can not be used for quick and correct orientation in the clinical condition of patients in the early stages of tumor growth.

Diffuse alveolar damage due to inappropriate strategy of mechanical ventilation in an experimental porcine model.

BACKGROUND: The aim of the study was to determine how tidal volumes may affect the lung and haemodynamics during mechanical ventilation. MATERIALS AND METHODS: With the approval of the Ethics Committee, the study included a total of 24 healthy piglets, average weight 30 kg (range 28-33 kg). The animals were ventilated for 90 minutes under general anaesthesia with two different tidal volume strategies and allocated into three groups. Group A, animals were healthy controls, Group B, animals comprised 8 piglets with an abdominal aortic aneurysm and ventilated with a low tidal volume strategy (VT(exp) 7 ml/kg). Group C comprised 8 animals seven days after kidney transplantation, ventilated with a high tidal volume strategy (VT(exp) 12 ml/kg). Changes in lung mechanics and hemodynamics were assessed at 30th and 90th minutes. Lung tissue samples were examined histologically. RESULTS: Protective ventilation in Group A and B did not confer any haemodynamic and lung mechanic differences. Significant differences were only found in Group C at 90 minutes for increased preload of both heart ventricles (CVP; t-test 4.07, p<0.01 and PAoP; t-test 8.43, p<0.01), pulmonary vascular resistance (t-test 3.11, p<0.05), and decreased expiratory tidal volume (t-test 6.07, p<0.01), dynamic lung compliance (t-test 3.83, p<0.01) and cardiac output (t-test 2.07, p<0.01). Diffuse alveolar damage was detected histologically. CONCLUSION: Mechanical ventilation at high tidal volumes reaching 12 ml/kg caused functional changes in the lungs, diffuse alveolar damage and reduction of cardiac output within 90 minutes.

Importance of miR-20a expression in prostate cancer tissue.

BACKGROUND: MicroRNAs (miRNAs), which are endogenously expressed regulatory noncoding RNAs, have an altered expression in tumor tissues. MiRNAs regulate cancer-related processes such as cell growth and tissue differentiation, and therefore, might function as oncogenes or tumor-suppressor genes. The aim of our study was to assess the expression of mir-20a, let-7a, miR-15a and miR-16 in prostate cancer (PCa) and benign prostatic hyperplasia (BPH) tissue and to investigate the relation between the expression of miRNAs and the clinicopathological features of PCa. PATIENTS AND METHODS: The study group comprised 138 patients: 85 patients with BPH and 53 patients with PCa. The total RNA was isolated from the tissue specimen core and miRNA expressions were quantified using a real-time RT-PCR method (TaqMan MicroRNA Assays). U6snRNA was used for the normalization of the miRNA expression. RESULTS: miR-20a expression was significantly higher in the group of patients with a Gleason score of 7-10 in comparison with the group of patients with a Gleason score of 0-6 (p=0.0082). We found no statistical differences in the miRNA expressions (mir-20a, let-7a, miR-15a and miR-16) in the PCa tissue samples in comparison with the BPH tissue samples. CONCLUSION: Our result shows that the more dedifferentiated PCa cells have a higher expression of miR-20a and this supports the oncogenic role of miR-20a in PCa carcinogenesis. The evaluation of miRNA expression could yield new information about PCa pathogenesis.

Differential display code 3 (DD3/PCA3) in prostate cancer diagnosis.

BACKGROUND: Early diagnosis of prostate cancer (PCa) in an organ-confined stage following radical treatment is the only potential curative approach in PCa. Prostatic-specific antigen (PSA) is very helpful in early diagnosis, but the main disadvantage is that it has a low positive predictive value in the range of the grey zone of 2.5-10 ng/mL, which results in a high number of needless biopsies. For this reason, new tests with better parameters are needed. One promising test is that for differential display code 3 (DD3(PCA3)), which is a prostate-specific non-coding mRNA that is highly overexpressed in prostate tumor cells. The aim of the present study was to evaluate the potential of DD3(PCA3) for mRNA in PCa diagnosis. PATIENTS AND METHODS: A total of 186 patients were examined. In a group of patients with suspected PCa, one tissue specimen core was collected for testing DD3(PCA3) expression. According to the histological verification there were 100 patients with benign prostatic hyperplasia, 12 patients with prostatic intraepithelial neoplasia and 74 patients with PCa. The total RNA was isolated and DD3(PCA3) and PSA expressions were quantified using quantitative RT real-time PCR method. The DD3(PCA3)/PSA mRNA ratio was determined for all groups. RESULTS: It was found that the levels of the mRNA expression of DD3(PCA3) were significantly higher (p<0.045) in patients with PCa than in patients with benign prostatic hyperplasia. No statistically significant differences in levels of mRNA expression of DD3(PCA3) between patients with organ-confined and those with advanced or metastatic disease, nor according to Gleason score, were found. CONCLUSION: DD3(PCA3) appears to be a promising marker for early detection of PCa and also for differential diagnosis between patients with benign prostate hyperplasia and those with PCa.