In-vitro antiplatelet effect of melatonin in healthy individuals and patients with type 2 diabetes mellitus.

PURPOSE: The incidence of acute myocardial infarctions (AMI) shows circadian variation typically peaking during morning hours with a decline at night. However, this variation does not occur in patients with diabetes mellitus (DM). The night's decline of AMI may be partially explained by melatonin-related platelet inhibition. Whether this effect is absent in diabetic patients is unknown. The aim was to study the effect of melatonin on in-vitro platelet aggregation in healthy individuals and patients with type 2 DM. METHODS: Platelet aggregation was measured in blood samples from healthy individuals (n = 15) and type 2 DM patients (n = 15) using multiple electrode aggregometry. Adenosine diphosphate (ADP), arachidonic acid (ASPI) and thrombin (TRAP) were used as agonists. Aggregability for each subject was tested after adding melatonin in two concentrations. RESULTS: In healthy individuals, melatonin inhibited platelet aggregation in both higher (10-5 M) and lower concentrations (10-9 M) induced by ADP, ASPI, and TRAP (p < 0.001, p = 0.002, p = 0.029, respectively). In DM patients, melatonin did not affect platelet aggregation in both concentrations induced by ADP, ASPI, and TRAP. Melatonin decreased platelet aggregation induced by ADP, ASPI, and TRAP significantly more in healthy individuals compared to patients with DM. (p = 0.005, p = 0.045 and p = 0.048, respectively). CONCLUSION: Platelet aggregation was inhibited by melatonin in healthy individuals. In-vitro antiplatelet effect of melatonin in type 2 DM patients is significantly attenuated.

Adipose-tissue-derived therapeutic cells in their natural environment as an autologous cell therapy strategy: the microtissue-stromal vascular fraction.

The prerequisite for a successful clinical use of autologous adipose-tissue-derived cells is the highest possible regenerative potential of the applied cell population, the stromal vascular fraction (SVF). Current isolation methods depend on high enzyme concentration, lysis buffer, long incubation steps and mechanical stress, resulting in single cell dissociation. The aim of the study was to limit cell manipulation and obtain a derivative comprising therapeutic cells (microtissue-SVF) without dissociation from their natural extracellular matrix, by employing a gentle good manufacturing practice (GMP)-grade isolation. The microtissue-SVF yielded larger numbers of viable cells as compared to the improved standard-SVF, both with low enzyme concentration and minimal dead cell content. It comprised stromal tissue compounds (collagen, glycosaminoglycans, fibroblasts), capillaries and vessel structures (CD31+, smooth muscle actin+). A broad range of cell types was identified by surface-marker characterisation, including mesenchymal, haematopoietic, pericytic, blood and lymphatic vascular and epithelial cells. Subpopulations such as supra-adventitial adipose-derived stromal/stem cells and endothelial progenitor cells were significantly more abundant in the microtissue-SVF, corroborated by significantly higher potency for angiogenic tube-like structure formation in vitro. The microtissue-SVF showed the characteristic phenotype and tri-lineage mesenchymal differentiation potential in vitro and an immunomodulatory and pro-angiogenic secretome. In vivo implantation of the microtissue-SVF combined with fat demonstrated successful graft integration in nude mice. The present study demonstrated a fast and gentle isolation by minor manipulation of liposuction material, achieving a therapeutically relevant cell population with high vascularisation potential and immunomodulatory properties still embedded in a fraction of its original matrix.

Fibrin-based delivery strategies for acute and chronic wound healing.

Fibrin, a natural hydrogel, is the end product of the physiological blood coagulation cascade and naturally involved in wound healing. Beyond its role in hemostasis, it acts as a local reservoir for growth factors and as a provisional matrix for invading cells that drive the regenerative process. Its unique intrinsic features do not only promote wound healing directly via modulation of cell behavior but it can also be fine-tuned to evolve into a delivery system for sustained release of therapeutic biomolecules, cells and gene vectors. To further augment tissue regeneration potential, current strategies exploit and modify the chemical and physical characteristics of fibrin to employ combined incorporation of several factors and their timed release. In this work we show advanced therapeutic approaches employing fibrin matrices in wound healing and cover the many possibilities fibrin offers to the field of regenerative medicine.

High serum carbonic anhydrase IX predicts shorter survival in head and neck cancer.

OBJECTIVES: The objective of the study was to investigate prognostic and predictive value of pretreatment soluble carbonic anhydrase IX (CA IX) blood serum concentration in patients with locally advanced head and neck cancer. BACKGROUND: Increased expression of CA IX in tumor tissues has been associated with treatment resistance and worth prognosis. Soluble form of CA IX, released from tumor cells, is detectable in blood serum and could be a convenient predictive factor of treatment effectiveness that would enable treatment individualization. METHODS: The prospective study evaluated 48 patients with locally advanced squamous cell carcinomas of head and neck, treated by radiotherapy or chemo-radiotherapy. Pretreatment soluble CA IX serum concentrations were examined using enzyme-linked immunosorbent assay. RESULTS: Soluble CA IX serum concentration failed to predict radiotherapy effectiveness in the studied patient population (p = 0.26). However, high CA IX serum concentrations have been associated with shorter overall survival (p = 0.035) CONCLUSION: High pretreatment CA IX serum concentration is a negative prognostic factor in locally advanced head and neck cancer patients (Tab. 1, Fig. 2, Ref. 23).

The relation between oxidative stress biomarkers and atrial fibrillation after pulmonary veins isolation.

INTRODUCTION: The current paradigm claims a link between oxidative stress and atrial fibrillation. The aim of our research was to study a relation between the percentage of time spent in atrial fibrillation (AF burden) and concentrations of oxidative stress biomarkers, before and after pulmonary veins isolation (PVI). METHODOLOGY: We included 19 patients (mean age 55±10years, 4 females and 15 males) with implanted loop recorders undergoing PVI. Plasmatic concentrations of advanced glycation end-products (AGEs), fructosamine, advanced oxidation protein products and thiobarbituric-acid reacting substances (TBARS) were measured and AF burden was recorded immediately before and 3months after the PVI. AF burden was also recorded 9months after the PVI. RESULTS: Post procedural AGEs concentration significantly negatively correlated with AF burden after 3months (ρ=-0.63; p<0.01) and 9months (ρ=-0.5; p=0.04), respectively as well as TBARS concentration significantly negatively correlated with AF burden after 9months (ρ=-0.61; p=0.01). CONCLUSION: Our study showed AGEs and TBARS to be potential predictors for AF burden after the PVI. We suppose that the more oxidative stress after the PVI is provoked, the more fibrotic tissue is produced. That means a better electrical isolation of pulmonary veins and consequently a lower AF burden.

Functional and circulatory renal changes in advanced heart failure.

OBJECTIVES: The aim of the study was to describe the relations between heart and renal functions and to investigate whether reduced glomerular filtration rate is influenced more by reduced perfusion or venous congestion. METHODS: A prospective cohort study of 101 patients (69 men, 32 women) with chronic heart failure aged 52 (49-54) (median, 95 % confidence interval) years. We analyzed the blood samples, parameters of echocardiography and right heart catheterization. RESULTS: Left ventricular ejection fraction correlated with the estimated glomerular filtration rate eGFR (r = 0.214, p = 0.036) in the whole sample of patients. We found a correlation between cardiac output and renal perfusion pressure in the whole sample (r = 0.232, p = 0.0225) and in patients with chronic heart failure (r = 0.254, p = 0.0278).In the whole sample the mean pulmonary artery pressure (PAP) correlated with the variables determining renal function: PAP and renal perfusion pressure (r = -0.345, p = 0.002),PAP and eGFR (r = -0.299, p = 0.009). In the other two studied groups these correlations were not significant. CONCLUSION: In the group of heart failure patients left and right ventricular functions were the main determinants for renal function. Current cardiac output or right atrial pressure as markers of renal perfusion were not associated with renal functions in advanced but stable heart failure patients with low burden of extracardiac comorbidities (Tab. 1, Fig. 4, Ref. 13).

[Relation between carbonic anhydrase IX serum level, hypoxia and radiation resistance of head and neck cancers]. / Vztah medzi sérovou hladinou karboanhydrázy IX, hypoxiou a rádiorezistenciou nádorov hlavy a krku.

BACKGROUND: Hypoxia of locally advanced head and neck cancers is one of the main causes of their radiation resistance that presents clinically as a persistence of residual tumor disease after radiation therapy. Therefore, detection of tumor hypoxia could be an important predictor of treatment efficacy. Carbonic anhydrase IX (CA IX) is a protein, coded by a homonymous gene, the expression of which increases in tumor tissues at hypoxic conditions. Hence, CA IX represents an endogenic marker of tumor hypoxia, identifiable in tumor tissues, and its soluble extracellular domain can also be detected in body fluids of the patient. The primary endpoint of this study was to explore whether a correlation exists between CA IX serum level and the residual tumor disease after therapy. The secondary endpoint was to find out how the serum concentration of CA IX changes during the course of fractionated radiation therapy. MATERIALS AND METHODS: The presented prospective monocentric clinical study evaluated a population of 30 patients with locally advanced squamous cell head and neck cancers, treated by radiation therapy or concurrent chemo radiation therapy with a curative intent. The serum concentration of the soluble form of CA IX was examined from a venous blood sample, using sandwich enzyme linked immunosorbent assay (ELISA). The blood samples were obtained before the treatment initiation, in the middle of radiation therapy, at the time of finishing radiation therapy and six weeks after the treatment completion. RESULTS: We found a substantial variability in the CA IX levels measured in the examined population, ranging 0- 1,696 pg/ ml. We found no significant changes in the mean value of CA IX concentration during the course of radiation therapy and after the treatment completion. In 11 patients (36.7%), the treatment resulted in complete remission of the disease. In these patients, lower average pretreatment levels of CA IX were noted when compared to patients with persistence of residual tumor disease (37.57 vs 77.47; p = 0.154). CONCLUSION: The results indicate that serum level of CA IX in patients with locally advanced head and neck cancers does not change significantly during the course of fractionated radiation therapy. The relation between CA IX serum level and residual tumor disease after radiation therapy requires verification on a larger population of patients.

Blood pressure impact on left ventricular geometry in chronic haemodialysis patients.

AIM: Left ventricular hypertrophy in chronic haemodialysis patients is multifactorial. Our aim was to evaluate retrospectively the relationship between 24-h blood pressure monitoring and geometry and function of left ventricle (LV). Patients a methods: We examined 50 patients (men/women 33/17) treated by chronic haemodialysis (>3 months) aged 57.5 years (53-63; median, interquartile range). We measured blood pressure during 24 hours in short interdialytic period using Spacelab monitor 90217. Echocardiography was provided in short interdialytic period. RESULTS: Left ventricular mass index significantly correlated with SBP (tau-b=0.21; p=0.030; 95%CI 0.01-0.42), DBP (tau-b=0.23; p=0.018; 95%CI 0.04-0.42) and MAP (tau-b =0.26; p=0.009; 95%CI 0.06-0.45). SBP, DBP, MAP and PP did add a significant information to the prediction of relative wall thickness. We did not find any relationship between BP and left ventricular ejection fraction, left ventricular enddiastolic diameter and left atrial size. CONCLUSION: We found out an important 24-hour blood pressure impact on left ventricular relative wall thickness and left ventricular mass index. Left ventricular ejection fraction, left ventricular enddiastolic diameter and left atrial size were not related to 24-hour blood pressure. We did not find a relationship between blood pressure and left ventricular enddiastolic diameter. From all diastolic parameters the strongest association was found between systolic blood pressure in all three phases and ratio of peak early to late diastolic filling velocity (Tab. 5, Ref. 19).

Short-term administration of alibernet red wine extract failed to affect blood pressure and to improve endothelial function in young normotensive and spontaneously hypertensive rats.

As wine polyphenols were shown to possess many positive effects in mammals, including improvement of vascular function, this study investigated the effect of the Slovak Alibernet red wine extract (AWE) on blood pressure and vascular function in young normotensive Wistar-Kyoto (WKY) and spontaneously hypertensive (SHR) rats. Six weeks old, male, WKY and SHR were treated with AWE for three weeks at the dose of 24.2 mg/kg/day. Blood pressure (BP), determined by tail-cuff plethysmography, was significantly elevated in SHR vs. WKY and AWE failed to affect it. Lipid peroxidation was evaluated by determination of thiobarbituric acid-reactive substances. Vascular function was assessed in rings of the femoral artery using Mulvany-Halpern's myograph. Maximal endothelium-dependent acetylcholine (ACh)-induced relaxation was reduced in control SHR vs. WKY rats by approximately 9.3 %, which was associated with a significant decrease of its NO-independent component. AWE failed to affect maximal ACh-induced relaxation, both its NO-dependent and independent components, compared to controls of the same genotype. AWE however reduced lipid peroxidation in the left ventricle of both WKY and SHR and in the liver of SHR. In conclusion, three-week administration of AWE failed to reduce BP and to improve endothelial function in the femoral arteries of both genotypes investigated.

Endothelial dysfunction in femoral artery of the hypertensive rats is nitric oxide independent.

This study examined nitric oxide (NO) production, oxidative load and endothelium-dependent relaxation (NO-dependent and NO-independent) in adult male borderline hypertensive (BHR) and spontaneously hypertensive (SHR) rats as compared to normotensive Wistar-Kyoto (WKY) rats. Systolic blood pressure (BP) was determined by tail-cuff. NO production was determined by conversion of [(3)H]-L-arginine. Conjugated dienes (CD) and concentrations of thiobarbituric acid-reactive substances (TBARS) were measured for assessment of oxidative load. Vascular function was investigated in rings of the femoral artery (FA) using a wire myograph. BP of WKY, BHR and SHR was 106+/-2, 143+/-3 and 191+/-3 mm Hg, respectively (p<0.01 for each). Significant left ventricle (LV) hypertrophy and elevated levels of CD and TBARS in the LV were present in BHR and SHR as compared to WKY. NO production was elevated significantly in the aorta of BHR and SHR vs. WKY as well as in the LV of SHR vs. WKY. Acetylcholine (ACh)-induced relaxation of the FA was reduced significantly in both BHR and SHR vs. WKY. The NO-dependent component of ACh-induced relaxation had increasing tendency in hypertensive groups and it correlated positively with BP. The NO-independent component of vasorelaxation was reduced significantly in BHR and SHR vs. WKY and it correlated negatively with BP. In conclusion, the results showed that endothelial dysfunction in the experimental model of borderline hypertensive and hypertensive rats is NO-independent. The results suggest that borderline hypertension represents a risk of other cardiovascular disorders which is qualitatively similar to that of fully developed hypertension.

[The p16INK4A mRNA transcripts estimation in cervical smear with different degrees of cervical dysplasia]. / Stanovenie expresie p16INK4A mRNA transkriptu v steroch z krcka maternice s rôznymi stupnami cervikálnej dysplázie.

OBJECTIVE: To determine the presence of HPV infection and expression level of p16INK4A mRNA transcripts in cervical smears as adjunct biomarker in detection of cervical intraepithelial neoplasia or cancer. DESIGN: Prospective pilot clinical study assessing clinical utility and validity of ddCt method for qPCR mRNA expression of p16ink4a in comparison to immunohistochemistry. SETTING: Department of Molecular Biology, Department of Obstetrics and Gynecology, Jessenius Medical Faculty, Commenius University, Martin, Slovak Republic. METHODS: Cervical smears (OC) from patients with different cervical lesions (L-SIL, H-SIL, SCA; n=45) and from healthy controls (n=45) were tested for the presence of HPV infection and p16INK4A mRNA transcripts using relative quantification (RQ). Results were compared to H&E and IHC histological findings from biopsies (conization, hysterectomy). RESULTS: HPV 16 was the most frequent finding (53.3%) in the group of subjects with cervical dysplasia. The p16INK4A mRNA expression analysis revealed the slightly reduced expression in L-SIL group, 4-fold increased expression in H-SIL and 10-fold increase in women with SCA when compared to controls. The p16INK4A mRNA expression in OC was present in 30% of L-SIL, 75% of H-SIL and 85.7% of SCA samples, respectively. The test overall sensitivity was 81.48% (95% CI: 61.92-93.7) and specificity 60% (95% CI: 26.24- 87.84) with PPV of 84.62% and NPV of 54.55%. The likelihood ratio (LR) in case of test positivity was 2.04 and for negativity 0.31. The diagnostic accuracy of p16INK4A expression by RQ method in OC smears for prediction of p16 positivity in cervical dysplasia was 66.7% for the L-SIL lesions, 59.5% for H-SIL lesions, and 100% for SCA (r=0.9897, p<0.0913) when compared to IHC p16 positive findings in surgically treated samples. CONCLUSION: The relative quantification is able to determine the level of p16INK4A mRNA transcripts in cervical smear cells with active carcinogenesis nearly at the same level as IHC staining. The advance of biopsy sparing over IHC is qualifying this diagnostic approach for useful candidate in selective management of women with cervical dysplasia looking for cervix preservation or avoiding the unnecessary overtreatment.

[Blood pressure changes in chronically haemodialysed patients]. / Zmeny krvného tlaku u chronicky hemodialyzovaných pacientov.

BACKGROUND: Poor blood pressure control in chronically haemodialysed patients leads to increased cardiovascular morbidity and mortality. Information on valid values of blood pressure during haemodialysis and out of office is very important in order to set up adequate treatment. AIM: To measure blood pressure during the haemodialysis and the subsequent 24-hour period using an ambulatory blood pressure monitoring (ABPM) in patients with normal blood pressure (BP) and patients with high normal BP and hypertension. Relationship between time-dependent blood pressure changes, ultrafiltration (UF) and interdialytic weight gain (IDWG) was analysed. PATIENTS AND METHODS: Fifty chronically haemodialysed (> 3 months) patients (males/females 33/18) aged 57.5 (53-63; median, interquartile interval) years were studied. Systolic and diastolic pressures (SP, DP) were measured during haemodialysis every hour (H0-H4) and over following 24 hours using Spacelab 90217 monitor. Pulse pressure (PP) values were calculated as a difference between SP and DP. The patients were stratified into two groups based on the cut-off-point calculated as the mean of two mean arterial pressure (MAP) values obtained at the beginning and after the first hour of HD: Group A (n = 25), MAP < 100 mm Hg; Group B (n = 25), MAP 100 mm Hg. Interdialytic weight gain was measured before HD (IDWG1) and after the ABPM (IDWG2); also ultrafiltration (UF) was obtained. The post-dialysis 24-h ABPM period was divided into eight 3-hour intervals (M1-8). RESULTS: During HD no significant change in SP, DP or PP was found in both group, but there was a significant difference (p = 0.01) between both groups in SP, DP and PP. Values of BP at the end of dialysis were in group A: SP 125 (120-130) mm Hg, DP 75 (60-80) mm Hg and PP 50 (40-60) mm Hg in group B: SP 150 (140-160) mm Hg, DP 80 (80-90) mm Hg a PP 60 (60-70) mm Hg. We did not find any influence of IDWG1 or IDWG2 on SP or DP in both groups. Relationship between UF 3 000 (2 500-4 300) ml and SP (Δ sTK -5 mm Hg) was confirmed only in group A (p = 0.04). In group A, we found a decrease in SP during the third and sixth 3-hour interval (p = 0.01; p = 0.02) including sleeping period, all compared to the end of HD (H4). In group B, such a decrease in SP was found only in the second sleep interval (p = 0.01) and in the sixth 3-hour interval (p = 0.03), all compared to the end of HD (H4). As to DP at the end of dialysis (H4) in group A, it differed only in the third 3-hour interval (p = 0.02), but not during the sleeping period. In group B, the decrease of DP compared to the end HD (H4) was recorded during the two sleep intervals (p = 0.01), and also in the sixth and seventh 3-hour intervals (p = 0.01; p = 0.03). In group A, PP was compared to the end of HD (PPH4) significantly decreased in the first 3-hour interval (p = 0.02) and in seventh and eight 3-hour interval (p = 0.03; p = 0.04). In group B, PP did not significant change from the end of HD. Difference in SP between both groups was maintained over the entire course of ABPM (p = 0.01). However, DP values in both groups were different in the first and third 3-hour intervals (p = 0.01) but in following intervals DP in group B decreased to the level of that in group A. There was no significant difference in the proportion of non-dippers and reverse dippers in both groups. CONCLUSION: Systolic, diastolic, mean arterial and pulse pressure pressures were not significantly changed during the haemodialysis in both groups. Relationship between ultrafiltration and systolic pressure was confirmed only in group A. No influence of interdialytic weight gain on blood pressure during 24 hours was seen in either group. Systolic pressure decreased in both groups during the nighttime compared to post-HD values, but diastolic pressure decreased only in group B. PP did not decrease during the night in any group. There was no significant difference in the proportion of non-dippers and reverse dippers in both groups.

[Assessment of vascular reactivity by wire myograph]. / Stanovenie cievnej reaktivity pomocou drôtikového myografu.

This paper describes the methodology and application of a wire myograph which has been used for the measurement of vascular reactivity. In an earlier years (pre-1970s) most of the information about the mechanical, morphological and pharmacological properties of vascular smooth muscle was confined only to larger arteries (mainly aorta). Whereas information about smaller arteries was purely inferred from perfusion experiments and histological examination. However, after mid-1970s Prof. Mulvany and Prof. Halpern developed and introduced an astonishing technique, a wire myography, to study the contractile responses of an isolated small resistance arteries (approximately 100-300 microm in internal diameter). This work describes some of the principles used in the investigation of the vessels, based on the use of the small vessel dual wire myograph. A dual myograph allows us simultaneous testing of two vessels. The technique allows segments of small arteries to be mounted as the ring preparations to the myograph chamber, and providing measurements of isometric responses. On the other hand, there are other techniques including an isobaric and isotonic mounting of arteries have been developed to date. The myograph has been used for the investigation of a variety of small and larger arteries and other tubular structures from a wide range of species. In the second part of this report we show an experimental example concerning measurement of endothelial functionality by technique described therein before.

Biophysical investigation on left ventricular myocytes in rats with experimentally induced diabetes.

Diabetes is a recognized risk factor of heart disease. The abnormalities related to a decreased heart performance probably arise at cellular and molecular levels already in the asymptomatic phase of diabetes. However, the early alterations initiating a sequence of events that culminates in the clinical signs have not been fully elucidated yet. This review deals with some biophysical methods applied to investigation of left ventricular myocytes in rats with streptozotocin diabetes, as well as our most important findings concerning diabetes-induced cell changes which cannot be captured by other techniques. The observed decrease in sarcolemmal membrane fluidity is causatively associated with increased glycation and glycoxidation. On the other hand, an increase in the mitochondrial membrane fluidity may be attributed to augmented energy transduction through the membranes. We reported for the first time concurrent measurements of membrane potential and dynamics, and respiratory chain activities in rat heart mitochondria, as well as calcium transients in the myocytes from diabetic hearts together with the assessed quantitative relationships among these variables. We were able to detect some significant alterations that may underlie myocyte dysfunction and subsequent remodeling of the heart. We suppose that not all these changes reflect mechanisms leading to pathology; some may represent adaptive and compensatory responses to diabetes.

No differences in outcome between radical surgical treatment (enucleation) and stereotactic radiosurgery in patients with posterior uveal melanoma.

To report the treatment outcome and possible survival difference between radical surgical treatment (enucleation) or stereotactic radiosurgery (SRS). LINAC stereotactic radiosurgery is an alternative treatment for posterior uveal melanoma used in Slovakia since 1999. The study analyzed patients treated for posterior uveal melanoma in the period 2001-2008. The aim of the study was to compare the relapse-free survival in the cohort of patients primarily treated with surgery (enucleation) or SRS. A total number of 84 patients were included, treatment was determined on a case-by-case basis. We reviewed the records of patients with ciliary body or choroidal melanoma treated by enucleation - 44 patients (52%) and SRS - 40 patients (48%). The therapeutic attitude was established on the basis of ophthalmoscopy, ultrasound (A, B mode), other ophthalmological findings, visual acuity, and general status of each patient. Volume of the tumor was calculated using the formula: "pi/6 x length x width x height" for each patient. All of the patients before decision to "conservative" attitude therapy underwent MRI examination. The therapeutic dose in SRS patients group was 35.0-38.0 Gy. The data were analyzed using Kaplan - Meier survival method for the differences in survival rates between the treatment groups, and afterwards by Cox s proportional hazard method with predictors involved. Among the baseline covariates evaluated, only age affected the prognosis for survival to a statistically important, however not significant degree. The risk of death among patients treated with enucleation relative to those treated with stereotaxy after adjustment for baseline characteristics of patients, age, and tumor volumes was not significant [1.82] (95% CI, 0.46 to 7.30; P = 0.396). The overall five-year survival rate for patients with posterior uveal melanoma was 72%. Treatment by either primary enucleation or SRS according to our results does not appear to influence the development of metastases in patients with uveal melanoma; the survival prognosis is essentially determined by the stage and character of the tumor. No survival difference attributable to stereotactic irradiation of uveal melanoma has been demonstrated in this retrospective study. A small difference is possible, but a clinically meaningful difference in mortality rates, whether from all causes or from metastatic melanoma, is unlikely. Treatment by either radical surgical attitude (enucleation) or "conservative" LINAC- SRS does not appear to influence the survival rate in patients with uveal melanoma.

Accurate normalization factor for wire myography of rat femoral artery.

Wire myograph is a device for the in vitro investigation of both, active and passive properties of arteries. Arteries from a variety of animal species, pathological states, and vascular beds were investigated using this method. We focus on the normalization procedure which is aimed to standardize experimental settings and, in part, to simulate physiological conditions. During normalization, it is determined the internal circumference of a vessel stretched to a tension that corresponds to the transmural pressure of 100 mm Hg (IC100). Once it is determined, the internal circumference is traditionally set to (0.9 IC100). However, this constant 0.9, called also the normalization factor (NF), was experimentally determined for rat small mesenteric arteries only. Therefore, the aim of our work was to show the influence of different NFs on the passive tension and reactivity of both, rat femoral arteries (FA) and the first branches of superior mesenteric arteries (MA). We found out that the maximal active wall tension of the FA was achieved at the NF value of 1.1, and that of the MA at 0.9. Considering the values of the active wall tension we suggest that higher reactivity and better signal-to-noise ratio in FA can be achieved when the NF is set at least to 1.0.

[Pre-operative care for cardiac surgery patients with cold antibody disorder, cryoglobulinaemia and cryofibrinogenemia]. / Péce a pacienty s nemocí chladovych protilátek, kryoglobulinemií a kryofibrinogenemií pred kardiochirurgickými zákroky.

We present an example of a patient with confirmed cold agglutinin disease who underwent cardiac surgery in hypothermia to illustrate a known fact that, when exposed to cold, cold agglutinins induce haemolysis of erythrocytes and that cryoglobulins and cryofibrinogens may, upon exposition to cold during a surgery under hypothermia, precipitate or gelify and thus increase plasma viscosity and damage microcirculation. Detailed immunological and haematological investigations in all patients awaiting cardiac surgery with a risk of developing hypothermia is not advantageous considering the low number of patients with clinical and laboratory signs of cold agglutinin disease, autoimmune haemolytic anaemia or paroxysmal cold haemoglobinuria and considering that these investigations, in addition, might not detect cryoglobulinaemia and cryofibrinogenemia. Identification of in-risk patients from the warning signs in the medical history, physical or basal laboratory testing who would subsequently undergo confirmatory investigations to verify the presence of these entities and define them accurately might be a potential solution to this clinical issue. Cardiac surgery strategy and peri-operative care should be tailored to the results of these investigations. Well-structured, practiced and functional cooperation between clinicians and laboratory personnel is a prerequisite for success in these circumstances.

Intramucosal bacteria in colon cancer and their elimination by probiotic strain Enterococcus faecium M-74 with organic selenium.

Intraepithelial bacteria were isolated by the gentamicin protection assay (GPA) from biopsy samples obtained at colonoscopy (colon cancer, n = 10 patients; colonic adenoma, n = 20; control group, n = 20; cancer patients without gastrointestinal tract GIT malignancy, n = 10). After a three-month administration of E. faecium M-74 to patients with positive GPA biopsies, 172 biopsy specimens from 60 patients were examined with the GPA. The number of biopsies with intracellular bacteria was significantly higher in adenoma and carcinoma group than in control group (26 vs. 10%; p = 0.004); in cancer patients without GIT malignancy the difference was nonsignificant. E. faecium M-74 was also administered to 5 patients with colonic adenoma; according to a control colonoscopy the number of biopsies with intracellular bacteria was significantly lower after probiotic administration (48 vs. 16%; p = 0.03). A striking prevalence of intraepithelial bacteria was also showed in patients with large bowel adenoma and carcinoma. The administration of probiotic strain M-74 can thus be considered to be an effective and promising method for elimination of pathogenic bacteria in the case of inflammatory bowel disease and colon cancer.