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Purpose: To examine the removal of pegylated liposomal doxorubicin (PLD) during plasmafiltration (PF) and determine whether the drug could be withheld prior to its organ distribution responsible for mucocutaneous toxicity., Methods: Six patients suffering from platinum-resistant ovarian cancer were treated with a 1-h IV infusion 50 mg/m(2) of PLD/cycle-for three cycles q4w. Over 44 (46)-47(49) h postinfusion, five patients (14 cycles in total) underwent PF using a cascade PF method consisted of plasma separation by centrifugation and plasma treatment using filtration based one volume of plasma treatment, i.e., 3.18 L (±0.6 L) and plasma flow 1.0 L/h (0.91-1.48 L/h). Doxorubicin concentration in blood was monitored by a high-performance liquid chromatography method for 116 h postinfusion. Pharmacokinetic parameters determined from plasma concentration included volume of distribution, total body clearance, half-life of elimination, and area under the plasma concentration versus time. The amount of doxorubicin in the body eliminated by the patient and via extracorporeal treatment was evaluated. Toxicity was tested using CTCAE v4.0., Results: The efficacy of PF and early responses to PLD/PF combination strategy were as follows: over 44(46) h postinfusion considered necessary for target distribution of PLD to tumor, patients eliminated 46 % (35-56 %) of the dose administered. Over 44(46)-47(49) h postinfusion, a single one-volume plasma filtration removed 40 % (22-45 %) (Mi5) of the remaining doxorubicin amount in the body. Total fraction eliminated attained 81 % (75-86 %). The most common treatment-related adverse events (grade 1-2) such as nausea (4/14 cycles-28 %) and vomiting (3/14 cycles-21 %) appeared during 44 h postinfusion. Hematological toxicity-anemia (5/14 cycles-35 %) was reported after cycle II termination. Symptoms of PPE-like syndrome (grade 1-2) appeared in one patient concomitantly with thrombophlebitis and malignant effusion. In this study, only one adverse reaction (1/14-7 %) as short-term malaise and nausea was reported by the investigator as probably related to PF., Conclusion: A single one-volume PF does remove a clinically important amount of doxorubicin in a kinetic targeting approach. There were no serious signs of drug toxicity and/or PF-related adverse events. Kinetically guided therapy with pegylated liposomal doxorubicin combined with PF may be a useful tool to the higher efficacy and tolerability of therapy with PLD.

Establishing translocated populations is a common process to preserve and maintain genetic diversity of threatened species. In 2001, three translocated populations of noble crayfish (Astacus astacus) were established in the Czech Republic, founded by either adult or juvenile individuals from three particular source populations. We assessed genetic diversity at seven microsatellite loci after one decade (assumed three generations) from establishment. Although the translocated populations exhibited a slight but non-significant reduction in genetic diversity (A R = 2.2-5.0; H O = 0.11-0.31), the most striking result was generally very low genetic diversity in source populations (A R = 3.0-5.3; H O = 0.15-0.38). Similarly, a high degree of inbreeding (F IS = 0.36-0.60) demonstrates the nature of source populations, already affected by isolation and small size. In spite of that, based on the results of this study, the establishment of new translocated noble crayfish populations was successful, since there is no significant decline in genetic variability and all populations are still viable. Although source populations did not exhibit high genetic diversity, their distinctiveness makes them possible to use for conservation purposes. Continued monitoring is necessary to track the long-term progress of the translocation program, including other parameters describing the state of the population, such as the occurrence and frequency of diseases or morphological changes.

Introduction: High incidence of colorectal cancer in the Czech Republic is an actual and demographically significant health issue. Half of all of the patients is older than 70 years. Both surgical and non-surgical treatment options in this group of patients depend on factors that are difficult to measure only by current oncological and anesthesiological classifications (cTcNcM, ASA). The objective of this paper is to measure the impact of age on the use of various treatment modalities within the protocol and their results, and also to suggest alternative options for therapy tolerance assessment., Methods: Analysis of data over a five-year period from the NOR database prepared by the Institute of Biostatistics and Analyses, Masaryk University., Results: In all parameters a difference was demonstrated between patients below the age of 70 and those above the age of 70 years. Older patients were disadvantaged. Only 11.2% of patients younger than 70 years were not treated, whereas 25.2% over the age of 70 years were not treated., Conclusion: A complex geriatric examination could improve the indication process in various treatment modalities, including surgery., Key Words: colorectal cancer - elderly - treatment - geriatric assesment.

Introduction: Liver metastases are diagnosed in 60% of patients with colorectal cancer, both at the time of diagnosis or later in the course of their management. Surgical treatment is the sole potentially curable method with 5-year overall survival of approximately 50%. However, only less than 20% of patients underwent liver resection. A joint effort of medical oncologists and surgeons is to increase the numbers of resected patients. The "liver first approach" (LFA) is one of approaches aimed at increasing resecability. The authors present their preliminary results using this method., Methods: 102 patients were included in the multicentre study supported by the grant IGA NT 13660 Evaluation of quality of multimodal treatment for patients with colorectal cancer liver metastases - conducted at the Central Military Hospital between September 2012 and January 2015. We used LFA in 12 patients (12%). Patients were indicated for liver resection based on good response to neoadjuvant systemic therapy. Multiple bilobar liver involvement (>4 metastases) was present in 11 cases and one large solitary metastasis in the right liver lobe in one case. The primary tumor was located in the rectum in 9 patients, in the rectosigmoid in 3 patients; 3 patients had a colostomy. Others showed no signs of bowel obstruction., Results: We have performed R0 resections in 11 cases, and two-stage hepatectomy with portal vein embolisation was indicated 3 times (in one case we did not finish the second stage due to quick progression after PVE). We performed major resections 7 times, along with sever extraanatomic resections, incl. 11 RFA (6 times in combination with major resections). Perioperative mortality was 0%, morbidity 33% (Dindo-Clavien >2). Ten patients underwent adjuvant chemotherapy, in 7 cases including radiotherapy of the small pelvis due to a local advanced primary tumor. Resection of the primary tumor was done in 7 patients (58%). Two patients died recently because of disease progression (17%); progression was observed in 6 patients (50%)., Conclusion: We deem the LFA suitable especially for patients with metastatic rectal tumors where adjuvant systemic therapy can be combined with radiotherapy. The timing of the resection of the primary tumor still remains a question: it is necessary to rule out potential recurrence of liver metastases, which affected more than 50% of the patients. The benefit of LFA must be confimed by randomised studies.

Lipoprotein apheresis (LA) is an effective treatment method the patients with severe hypercholesterolemia, resistant to the standard therapy. LA is an extracorporeal elimination technique, which specifically removes low density lipoprotein (LDL) cholesterol from the circulation. At present, lipoprotein apheresis, combined with high-dose statin and ezetimibe therapy, is the best available means of treating patients with homozygous and statin refractory heterozygous familial hypercholesterolaemia (FH). However, the extent of cholesterol-lowering achieved is often insufficient to meet the targets set by current guidelines. The recent advent of new classes of lipid-lowering agents provides new hope that the latter objective may now be achievable. These compounds act either by reducing low density lipoprotein (LDL) cholesterol production by inhibiting apolipoprotein B synthesis with an antisense oligonucleotide (mipomersen), or by inhibiting microsomal triglyceride transfer protein (lomitapid), or by enhancing LDL catabolism via monoclonal antibody-mediated inhibition of the activity of proprotein convertase subtilisin/kexin 9 (PCSK9-alirocumab, evolocumab etc). The promising is the combination of LDL-apheresis with new drugs, namely for its potential to further decrease of LDL-cholesterol between apheresis. Depending on the outcome of current trials, it seems likely that these compounds, used alone or combined with lipoprotein apheresis, will markedly improve the management of refractory FH.

Background: Careful interpretation of discordant results in high-sensitivity troponin measurements is necessary in cases of suspect immunoassay interferences. We describe several procedures taken in a case of a polymorbid patient with chest pain, without clear evidence of myocardial necrosis and with increased high-sensitivity cardiac troponin T (hs-cTnT). We checked the Vafaie's algorithm for the evaluation of suspect interference in troponin measurements., Methods: We conducted a case report analysis, additional measurements, a dilution test and pretreatment of plasma with blocking agents., Results: Concentration of hs-cTnT (99 th percentile of "healthy" population 14 ng/L) increased from 120.1 ng/L to 280.4 ng/L during an 8-month period and decreased to 216.3 ng/L during the following month with repeatedly negative troponin I (TnI), hs-cTnI, myoglobin and creatine kinase MB (CK-MB). Suspected false positivity of hs-cTnT was further confirmed by treatment of plasma with an antiheterophile blocking agent (hs-cTnT before treatment 280.4 ng/L, after 16.53/16.23 ng/L). This outcome was further confirmed by the manufacturer's experiments., Conclusions: The false-positive results of hs-cTnT were caused by the presence of extremely rare high molecular weight protein, presumably IgM, most likely HAMA (human anti-mouse antibody). Only the pre-treatment of plasma with a blocking agent provided a reliable indication of the interference. Cooperation among clinicians, laboratory personnel and the manufacturer is essential., (Copyright © 2015 Elsevier B.V. All rights reserved.)

One of the generally recognized factors contributing to the initiation and maintenance of atrial fibrillation (AF) is structural remodeling of the myocardium that affects both atrial cardiomyocytes as well as interstitium. The goal of this study was to characterize morphologically and functionally interstitium of atria in patients with AF or in sinus rhythm (SR) who were indicated to heart surgery. Patient population consisted of 46 subjects (19 with long-term persistent AF, and 27 in SR) undergoing coronary bypass or valve surgery. Peroperative bioptic samples of the left and the right atria were examined using immunohistochemistry to visualize and quantify collagen I, collagen III, elastin, desmin, smooth muscle actin, endothelium and Vascular Endothelial Growth Factor (VEGF). The content of interstitial elastin, collagen I, and collagen III in atrial tissue was similar in AF and SR groups. However, the right atrium was more than twofold more abundant in elastin as compared with the left atrium and similar difference was found for collagen I and III. The right atrium showed also higher VEGF expression and lower microvascular density as compared to the left atrium. No significant changes in atrial extracellular matrix fiber content, microvascular density and angiogenic signaling, attributable to AF, were found in this cohort of patients with structural heart disease. This finding suggests that interstitial fibrosis and other morphological changes in atrial tissue are rather linked to structural heart disease than to AF per se. Significant regional differences in interstitial structure between right and left atrium is a novel observation that deserves further investigation.

Cherax (Astaconephrops) gherardii n. sp. is a moderate burrowing crayfish endemic to the Ajamaru Lakes of West Papua, Indonesia. This species is one of the crayfish species from this region that are exploited for ornamental purposes. Its commonly used commercial name in the pet trade is "Rainbow Crayfish" or "Blue Moon Crayfish", and its native name is "udang kuku biru". The new species is genetically and morphologically similar to Cherax boesemani, however, both species may be easily distinguished morphologically or by using sequence divergence, which is substantial for considering C. gherardii n. sp. to be a valid species.

Ultrastructure of spermatozoa of redclaw Cherax quadricarinatus and yabby Cherax destructor were described and compared. The acrosome complex and nucleus are located at the anterior and posterior region of the spermatozoon, respectively. The acrosome is a complex vesicle divided into two parts: the main body of the acrosome appears as a dense cup-shaped structure in longitudinal sagittal view, with the subacrosome zone occupying the central area of the vesicle. The acrosome is larger in C. quadricarinatus (width 2.37±0.27μm, length 1.31±0.23μm) than in C. destructor (width 1.80±0.27μm, length 1.01±0.15μm). There was no significant difference in L:W ratios of the studied species. The subacrosome zone in both species consists of two areas of different electron density. The nucleus is substantially decondensed and irregular in shape, with elaborate extended processes. The examined species exhibited a well-conserved structure of crayfish spermatozoon, similar to those of Cherax cainii and Cherax albidus. Small acrosome size, the absence of radial arms, and an extracellular capsule seem to be the morphological features that mostly distinguish Cherax from the Astacidae and Cambaridae., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Aim. To evaluate the long-term effect of rheohemapheresis (RHF) treatment of age-related macular degeneration (AMD) on photoreceptor IS/OS junction status. Methods. In our study, we followed 24 patients with dry AMD and drusenoid retinal pigment epithelium detachment (DPED) for a period of more than 2.5 years. Twelve patients (22 eyes) were treated by RHF and 12 controls (18 eyes) were randomized. The treated group underwent 8 RHF standardized procedures. We evaluated best-corrected visual acuity, IS/OS junction status (SD OCT), and macular function (multifocal electroretinography) at baseline and at 2.5-year follow-up. Results. RHF caused a decrease of whole-blood viscosity/plasma viscosity at about 15/12%. BCVA of treated patients increased insignificantly (P = 0.187) from median 74.0 letters (56.2 to 81.3 letters) to median 79.0 letters (57.3 to 83.4 letters), but it decreased significantly from 74.0 letters (25.2 to 82.6 letters) to 72.5 letters (23.4 to 83.1 letters) in the control group (P = 0.041). The mfERG responses in the region of eccentricity between 1.8° and 7° were significantly higher in treated patients (P = 0.04). Conclusions. RHF contributed to sparing of photoreceptor IS/OS junction integrity in the fovea, which is assumed to be a predictive factor for preservation of visual acuity.

Ovarian cancer is the fifth most common malignancy in the world's female population and with the highest lethality index among gynecological tumors. The prognosis of metastatic disease is usually poor, especially in platinum-resistant cases. There are several options for the treatment of metastatic disease resistant to platinum derivates (e.g. paclitaxel, topotecan and pegylated liposomal doxorubicin), all of which are considered equipotent. Pegylated liposomal doxorubicin (PLD) is a liposomal form of the anthracycline antibiotic doxorubicin. It is characterized by more convenient pharmacokinetics and a different toxicity profile. Cardiotoxicity, the major adverse effect of conventional doxorubicin, is reduced in PLD as well as hematotoxicity, alopecia, nausea and vomiting. Skin toxicity and mucositis, however, emerge as serious issues since they represent dose and schedule-limiting toxicities. The pharmacokinetics of PLD (prolonged biological half-life and preferential distribution into tumor tissue) provide new possibilities to address these toxicity issues. The extracorporeal elimination of circulating liposomes after PLD saturation in the tumor tissue represents a novel and potent strategy to diminish drug toxicity. This article intends to review PLD characteristics and the importance of extracorporeal elimination to enhance treatment tolerance and benefits.

Lipid apheresis (extracorporeal lipoprotein elimination) is administered to patients with familial hypercholesterolemia who fail to respond to standard therapy. The nature of the treatment process raises the suspicion that it decreases not only cholesterol but also antioxidants. A group of 12 patients (average age 47±17 y, 4 homozygous and 8 heterozygous individuals) with familial hypercholesterolemia treated by LDL-apheresis or rheohaemapheresis for 3-12 y was included in the study. In addition to cholesterol and triacylglycerol levels, vitamin E and vitamin A and also other markers of antioxidant activity were investigated. Nevertheless, the most important determined parameter was the vitamin E/cholesterol ratio in serum and lipoproteins. The results indicate that both extracorporeal elimination methods are effective and suitable ways to treat severe familial hypercholesterolemia, as the LDL fraction of cholesterol decreased by approximately 77% and 66% following LDL-apheresis and rheohaemapheresis, respectively. In addition, the serum vitamin E decreased by 54% and 57% and the decrease of the serum vitamin A was approximately 20%. However, the main marker of antioxidant capacity, vitamin E/cholesterol ratio, in the serum, VLDL and LDL significantly increased. The increase of vitamin E levels in the erythrocyte membranes of 2% following LDL-apheresis and a significant increase of 4% following rheohaemapheresis were confirmed. The presented results indicate that LDL-apheresis and rheohaemapheresis can be considered to be safe procedures according to the antioxidant capacity of the serum, VLDL and LDL lipoprotein fractions and the erythrocyte membrane.

LDL-apheresis is an extracorporeal elimination technique, which specifically removes LDL-cholesterol from the circulation. There are six methods for the selective LDL-cholesterol removal these days. The main indications for LDL-apheresis are the diagnosis of homozygous familial hypercholesterolemia, heterozygous familial hypercholesterolemia which is refractory the standard care and intolerance of routine care, and also patients with lipoprotein(a) increase resistant to the farmacotherapy. There is still debate which LDL-cholesterolemia is indication for LDL-apheresis therapy, and the recommendation differs among various countries. Despite large randomized trials are missing, there are several good quality studies to conclude, that the beneficial cardiovascular effects of LDL-apheresis in severe hypercholesterolemia are important and beneficial.

Assessment of the treatment procedures and their results is increasingly important in current medicine. The emphasis is put on an effective use of the health technologies (HTA). Unlike randomised studies, which involve strictly selected groups of patients who meet inclusion and exclusion criterias, the multicentre clinical registries provide a real-life picture of the treatment safety and effectiveness. Well prepared registries involve both research database and a friendly user interface enabling collection of parametric and easily analyzable data. Although there are some technological aspects aiming to ensure a maximum quality of entered data, cooperation with the users and data managers is essential. Such a registry, otherwise meaningful, must provide answers to previously defined medical hypotheses. Regular feedback to users (so called benchmarking or reporting) is considered to be of key importance. The Czech TAVI Registry (CTR) is a good example of reaching all of the above defined criterias. This registry contains data of approximately 95 % of all transcatheter aortic valve implantations (TAVI) performed in the Czech Republic. It is based on a general system aimed at the design of clinical trials, namely the TrialDB2 (a database system for clinical registries developed by the Institute of Biostatistics and Analyses at the Masaryk University (IBA MU). CTR has been run as an English-language version under the auspices of the Czech Society of Cardiology and represents one of the top-quality registries maintained by IBA MU. This paper presents the currently available database systems and some reports from this particular registry.

Introduction: Patients with serious aortic stenoses treated by conservative therapy have significantly worse life expectancy. Besides the surgical aortic valve replacement (AVR) as the gold standard of therapy, the transcatheter aortic valve implantation (TAVI) is indicated in patients at very high risk or who are contraindicated to AVR. The role of learning-curve in TAVI has to be established., Aim: Assessment of the results of consecutive TAVI procedures based on the experience of the team., Methods: 58 high-risk consecutive patients with the average age of 82.2 years were divided into 3 groups based on the TAVI order ( 20., 21.-40., 41.-58.). After the Edwards SAPIEN implantation via transfemoral or transapical approaches, all patients were followed for minimum 30 days. Data from the national registry (Czech TAVI Registry) were used for the retrospective analysis. Comparison of the groups was done by using the Kruskal-Wallis test, Mann-Whitney U test and χ2 test., Results: Significantly shorter procedural time (p < 0.001), hospitalization (p = 0.033) and a lower amount of contrast medium (p < 0.001) was observed during the time. There was no difference in the rate of clinical complications at 30 days., Conclusions: Increasing experience of the TAVI implantation team is associated with significantly shorter procedural time, hospitalization and a lower amount of contrast medium. Overall very good clinical results during 30 days were not affected by the team experience.

In January 2014, a programme of personalised invitations was launched in the Czech Republic, with the objective of inviting insured persons to cancer screening programmes; namely breast cancer screening and cervical cancer screening in women, and colorectal cancer screening both in women and men. This programme aims at strengthening the current cancer prevention programmes, and to increase the currently inadequate participation of the target population in these programmes; therefore, personalised invitations are sent to citizens who have not participated in these programmes for several years and therefore at risk of developing a serious disease. The project is coordinated by the Czech Ministry of Health in cooperation with the expert medical societies involved (gynaecology, gastroenterology, gastrointestinal oncology, diagnostic radiology, general practice), representatives of health care payers, and other experts nominated by the Minister of Health. All health care payers invite their clients (insured persons) to preventive check-ups, covering all examinations needed. The project has been realised with the assistance of financial resources from EU funds. This article describes the methodology of personalised invitations which has been implemented nationwide, its data background, and the first results of the project in the first half of 2014, when almost 1.3 million Czech citizens were invited.

Background: Rheohemapheresis (RHF) is a method that can stop the activity of the dry form of age-related macular degeneration (AMD). The pathophysiologic mechanisms are not well understood, and the effects of the RHF procedures extend beyond the time of the individual procedures., Patients and Methods: We present the data for 46 patients with AMD treated with a series of 8 rheohemapheretic procedures. Blood count parameters were measured before the first and the last procedures. The clinical effect was judged by changes in the drusenoid pigment epithelium detachment (DPED) area before and after the rheopheretic sessions., Results: Rheopheresis caused a decrease in hemoglobin (P<0.001), a decrease in leukocytes (P<0.034), and an increase in platelets (P<0.005). We found a negative correlation between the amount of platelets and their volume (P<0.001, Pearson correlation coefficient: -0.509). We identified the platelet/MPV ratio as a good predictor of the clinical outcome. Patients with a platelet/MPV ratio greater than 21.5 (before the last rheopheresis) had a significantly better outcome (P=0.003, sensitivity of 76.9% and specificity of 80%)., Conclusion: Several basic blood count parameters after RHF can be concluded to significantly change, with some of those changes correlating with the clinical results (reduction of the DPED area).

Czech cancer screening programmes feature a comprehensive multimodal information system which covers all the levels of assessment needed - population-based monitoring (Czech National Cancer Registry), monitoring of results in the diagnostic databases of centres, as well as the quantification of coverage and outputs of primary care according to the administrative data of health care payers. A system of personalised invitations to cancer screening programmes was launched in 2014, based on a stand-alone component of the information system which makes it possible to identify eligible clients in health care payers databases. The system was fully standardised and uniformly implemented in all health insurance companies; its functionalities also involve both continuous and retrospective assessment of the results of personalised invitation. The legislative framework in force will have to be applied and implemented for a more comprehensive and integrated employment of all involved data sources, i.e. cancer registries, screening registries, and administrative data. The system must be able to analyse de-identified individual records on clients participation in screening programmes, and thus to ensure an adequate analysis of performance indicators in compliance with international recommendations.

Background: As a part of the development of a new prospective payment model for radiotherapy we analyzed data on costs of care provided by three comprehensive cancer centers in the Czech Republic. Our aim was to find a combination of variables (predictors) which could be used to sort hospitalization cases into groups according to their costs, with each group having the same reimbursement rate. We tested four variables as possible predictors -  number of fractions, stage of disease, radiotherapy technique and diagnostic group., Methods: We analyzed 7,440 hospitalization cases treated in three comprehensive cancer centers from 2007 to 2011. We acquired data from the I COP database developed by Institute of Biostatistics and Analyses of Masaryk University in cooperation with oncology centers that contains records from the National Oncological Registry along with data supplied by healthcare providers to insurance companies for the purpose of retrospective reimbursement., Results: When comparing the four variables mentioned above we found that number of fractions and radiotherapy technique were much stronger predictors than the other two variables. Stage of disease did not prove to be a relevant indicator of cost distinction. There were significant differences in costs among diagnostic groups but these were mostly driven by the technique of radiotherapy and the number of fractions. Within the diagnostic groups, the distribution of costs was too heterogeneous for the purpose of the new payment model., Conclusion: The combination of number of fractions and radiotherapy technique appears to be the most appropriate cost predictors to be involved in the prospective payment model proposal. Further analysis is planned to test the predictive value of intention of radiotherapy in order to determine differences in costs between palliative and curative treatment.

Purpose. Determining long-term effects of rheohaemapheresis on the dry form of age-related macular degeneration. Methods. This study evaluates 19 patients, average age of 67.6 years, treated with rheohaemapheresis and 18 patients, average age of 72.8 years, comprising the control group. Minimum follow up period was 3.5 years. Each treated patient received a series of 8 sessions of rheohaemapheresis of 1.5 plasma volumes within 10 weeks. We measured the drusenoid pigment epithelium detachment (DPED), best-corrected visual acuity (BCVA), electroretinography (ERG), and rheological parameters. Results. In the treatment group, the baseline BCVA was 0.74 (0.36-1.0) 95% CI and BCVA after 3.5 years was 0.79 (0.41-1.0) 95% CI (P = 0.726). In the control group, the baseline BCVA was 0.71 (0.15-1.0) 95% CI and BCVA after 3.5 years decreased to 0.7 (0.32-0.87) 95% CI (P = 0.031). Baseline DPED was 6.78 ± 3.79 mm(2); after 3.5 years, it decreased to 4.13 ± 3.84 mm(2) (P < 0.001). In the control group, the baseline DPED was 4.09 ± 3.48 mm(2); after 3.5 years, it increased to 6.69 ± 4.2 mm(2) (P = 0.001). We noted increasing levels of positive wave peaking at 50 milliseconds (P50) after treatment (P = 0.022) and a stable amplitude of photopic responses of treated patients. Conclusion. Over the long term, rheohaemapheresis reduced the DPED, improved the function of photoreceptors, and prevented the decline of BCVA.

Introduction: Autologous stem cell transplantation (ASCT) became standard of care for patients with multiple myeloma (MM) under the age of 65 years. We routinely perform ASCT for newly diagnosed MM since 1996 in our department., Patients and Methods: We retrospectively analyzed all 285 transplants in 185 patients done for MM from January 1996 till December 2010. We analyzed overall survival (OS) and progression-free survival (PFS) regarding conditioning, stage, complete or very good partial remission (CR, VGPR) achievement, renal impairment, single vs. double transplant., Results: Estimated 10-years survival of the whole set of patients is 39% (median survival 95 months). Patients with renal impairment show same OS as those without (p = 0.22). Patients show similar overall survival and event free survival regardless of type of transplant. We observed better outcome in terms of overall survival in patients treated with new drugs (p = 0.0014). Reaching CR or VGPR was not translated into better OS (p = 0.30) and EFS (p = 0.10). Also stage of the disease and whether single or double transplant was used did not make any significant difference in the outcome., Conclusion: Stem cell transplantation greatly improved outcome of patients with MM. Poor outcome of allogeneic transplantation in our group of patients is related to high transplant related mortality (20% vs. 0%) and unexpected high relapse rate. There is a trend towards better survival, when new drugs are incorporated at any time in the course of the disease. This fact supports hypothesis that use of these drugs with ASCT should translate into better long-term outcome.

Cell mobilization, a process that influences circulation, margination, and finally, homing play key roles in the regeneration processes mediated by stem cells. Recent studies as well as prior studies from our group indicate an important role of the spleen in hematopoietic reconstitution, but to date the role of the spleen in hematopoietic reconstitution has been unclear and it has not been precisely documented in ablated animals. Therefore, we undertook the present study to define more closely the role of the spleen in hematopoietic reconstitution in lethally irradiated mice. After transplantation of irradiated mice with lacZ+ -marked lin- / CD117+ bone marrow cells, we compared splenectomized mice (T(S), splenectomy performed prior to irradiation) to nonsplenectomized, irradiated mice (T(N)) as well as to normal (unirradiated) mice. Impaired hematopoietic reconstitution was observed in T(S) mice. Splenectomy markedly altered the distribution of hematopoietic stem cells, as demonstrated by fluorescence-activated cell sorting analysis of endogenous CD117+ cells in the thymus and bone marrow of recipients. Cell engraftment was demonstrated by histochemical and polymerase chain reaction analyses of recipient tissues. These experiments demonstrated that in T(S) animals, transplanted hematopoietic stem cells mobilized to extravascular tissues, particularly the gastrointestinal tract. The number of donor cells in recipient tissues continued to increase for 30 days after transplantation with the highest numbers observed in the T(S) group. DNA marking analysis led to the conclusion that engrafted cells were not only integrated into recipient tissues but were also capable of performing complex cellular processes, including proliferation and repair. Our results are consistent with the novel possibility that cellular repair markedly affects stem cell regenerative functions and that repair is markedly influenced by the integrity and presence of organs not directly involved in specific tissue regeneration processes, particularly the spleen.

Background: The aim of our study was to compare healthcare-related costs of treatment with XELOX and FOLFOX4 chemotherapeutic regimens in patients with colorectal cancer. We have evaluated costs claimed to the health insurance by the hospital administering these cancer therapies. This study is a pilot project utilising the new I-COP database developed by the Institute of Biostatistics and Analyses of the Masaryk University in Brno, Czech Republic., Patients and Methods: First, we estimated the costs based on current prices of procedures, medication, and materials from public sources. Using the I-COP database, we then carried out a matched-pair comparison of 26 patients treated with FOLFOX4 or XELOX for colorectal cancer. We evaluated a period of three months of therapy (i.e. 6 cycles of FOLFOX4 or 4 cycles of XELOX). Statistical analysis was done using the Wilcoxon matched pairs test., Results: The estimated cost for three months of therapy was 148,288 Czech crowns (CZK) for FOLFOX4 (including CZK 101,064 for chemotherapy drugs) and CZK 123,756 for XELOX. The overall costs claimed to the insurance companies were CZK 160,158 and CZK 151,176 for FOLFOX4 and XELOX, respectively (p = 0.221). The XELOX regimen had significantly higher costs for chemotherapy drugs (CZK 131,705 versus 114,531, p = 0.023) whereas other costs were lower than those for FOLFOX4., Conclusions: FOLFOX4 and XELOX regimens can be considered as equivalent in terms of costs claimed by the hospital administering cancer treatment.

Introduction: Toxic epidermal necrolysis (TEN) is a rare, life-threatening disease with a high mortality rate that is linked to drug toxicity. There is a lack of data about the underlying pathophysiologic mechanisms and treatment options. The only widely accepted treatment of TEN is withdrawal of the offending drug followed by supportive care. The potential roles of corticosteroids, intravenous immunoglobulin (IVIG) and plasmapheresis (TPE) remain controversial., Aims: We present four patients with severe TEN (all with >80% involvement of body surface) who were treated with TPE following unsuccessful treatment with corticosteroids/IVIG., Methods: TPE was performed using a COBE Spectra blood cell separator. ACD-A was used as anticoagulant fluid and the target-washed plasma volume was one body volume. Plasma was replaced by a 5% solution of human albumin + Ringer's lactate., Results: The mean number of TPE sessions was 5.25 ± 2.22 (range 3-8). Drugs were implicated as an etiologic agent in each case. TPE led to prompt improvement of acute condition and general health as well as halting of disease progression. Additionally, the restoration of the epithelium began in all four patients., Conclusion: Plasmapheresis should be considered as an alternative treatment modality for patients with the most severe form of TEN if initial treatment with other agents, including corticosteroids and/or IVIG, fails. Drugs were suspected to be the cause of TEN in all four cases., (Copyright © 2012 Wiley Periodicals, Inc.)

To date, rheological treatment is the only chance to control the advanced dry form of age-related macular degeneration and arrest its progression to legal blindness. Rheohaemapheresis can change the main rheological parameters, blood and plasma viscosity, as well as change erythrocyte aggregability, improve erythrocyte flexibility and lead to substantial improvement when other methods of therapy fail. In this study, we describe changes in the levels of rheological efficacy indicators after rheohaemapheresis and their clinical significance in the dry form of age-related macular degeneration (AMD). Seventy-two patients with AMD were randomised; 34 controls, and 38 patients were treated with rheohaemapheresis (separator Cobe Spectra + Evaflux filter). After the procedures, α2-macroglobulin levels decreased by approximately 58%, fibrinogen by approximately 65%, IgM by approximately 67%, LDL cholesterol by approximately 71%, apolipoprotein B by approximately 65%, and lipoprotein (a) by approximately 42%. These decreases correspond with a decrease in blood and plasma viscosity (14/12%), clinical improvement (arrest of disease progression, even visual improvement in some cases), and heretofore-unreported improvement (even reattachment) of drusen retinal pigment epithelium detachment. Our modification of rheohaemapheresis is safe (5.4% of patients experienced clinically insignificant side effects).

Background: The current treatment of hereditary hemochromatosis (HH) consists of performing periodic whole blood phlebotomies. Erythrocytapheresis (EA) can remove up to three times more red blood cells per single procedure and could thus have a clinical benefit. A prospective study of 30 consecutive cases of HH were included in a periodic EA program., Methods and Patients: EA were performed using a discontinuous flow cell separators. The protocol consisted of a bimonthly EA until normalization of the serum ferritin was reached. The aim was to reduce the total erythrocyte volume by 25-35%, eventually, to adjust the amount so that hematocrit would not drop below 0.25., Results: 530 +/- 101 ml of erythrocytes were removed (median 517, range 116-761 ml). Iron depletion (ferritin < 20 microg/l) was achieved in all patients after a mean 6.9 +/- 7.6 months, median 5 months, range 1-36 months and a mean 14 EA sessions. The procedures were well tolerated and there were no severe side-effects., Conclusions: We conclude that HH patients treated with EA achieved iron depletion quickly under good conditions of tolerance. The efficacy, speed, tolerability, and more favorable schedule of an EA program facilitate treatment of HH.

Purpose: To evaluate the influence of haemorheopheresis on anatomical and functional findings in patients with soft-drusen maculopathy., Methods: We investigated 29 eyes (16 patients) and randomized 25 eyes (16 controls) with soft-drusen maculopathy [soft, confluent and reticular drusen, drusenoid retinal pigment epithelium detachment (RPED)]. Each patient received a series of eight haemorheophereses (cascade filtration of 1.5 plasma volume) within 10 weeks. The patients were followed up using Early Treatment Diabetic Retinopathy Study (ETDRS) charts, optical coherence tomography, fluorescein angiography, electroretinography and measurements of pulsed ocular blood flow., Results: After the procedures, there was a substantial reduction in rheologically active substances [lipoproteins, α2-macroglobulin, immunoglobulin M (IgM), fibrinogen], plasma and blood viscosity. At the 1.5-year follow-up, we noticed soft drusen absorption; reattachment of drusenoid RPED and stabilization or improvement of visual acuity occurred in 72% of patients in comparison to only 39% of patients in the control group. Full-field electroretinograms showed significantly higher scotopic activity of treated patients in comparison with the control group, and mainly insignificant differences in photopic activity between both groups. Despite the significant increase of activity in the paramacular retina in treated patients, the differences in amplitudes of multifocal electroretinography (mfERG) average responses were insignificant between groups., Conclusion: Haemorheopheresis seems to be capable of changing the activity of promoters of the natural course of soft-drusen maculopathy, its development and progression. Visual acuity and electrical activity of the retina can be stabilized or even improved. The therapy has been shown to be effective and safe., (© 2010 The Authors. Journal compilation © 2010 Acta Ophthalmol.)

Atorvastatin belongs to the group of lipid-lowering drugs known as statins. They significantly reduce the levels of total cholesterol, low-density cholesterol and plasma triglycerides therefore they are widely used in the treatment of hypercholesterolemia. Recently developed methods for the determination of atorvastatin and its metabolites in plasma used SPE (solid phase extraction) or LLE (liquid-liquid extraction) as the sample preparation step. However, both procedures are quite time-consuming and need relatively high volume of solvent/sample, which is impractical for the routine analyses of many biological samples. The aim of this work was to develop and validate more suitable sample preparation method for the determination of atorvastatin and its metabolites in biological samples using MEPS (microextraction by packed sorbent). The optimal conditions of MEPS extraction were using C8 sorbent and only 50 μl of the sample. The analytes were eluted by 100 μl of the mixture of acetonitrile:0.1 M ammonium acetate pH 4.5 (95:5, v:v). The analytical method was validated and demonstrated good linearity (r(2)>0.9990), recovery (89-115%) and intra-day precision (RSD<10%). Total time of the sample preparation was three times shorter (7 min) compared to SPE. The volume of sample was twenty times lower and the volume of solvents about ten times lower compared to SPE. Combination of fast MEPS method together with quick UHPLC-MS/MS was used for the determination of atorvastatin and its two metabolites in serum obtained from patients with familiar hypercholesterolemia., (Copyright © 2011 Elsevier B.V. All rights reserved.)

In this report, we describe the homing of hematopoietic stem cells (HSCs) to non-hematopoietic tissues in lethally irradiated (9Gy) hybrid mice transplanted intravenously with lin(-)/CD117(+) bone marrow cells from ROSA26 mice. The numbers of CFU-GM in spleen of irradiated transplanted mice were well above the levels found in non-irradiated group as early as day 8 after transplant. On 12th day regeneration of lymphocytes was observed, an increase in granulocytes was detected as late as on 33rd day. Transplanted cells containing lacZ gene were detected in recipient mice by histochemistry and their location in the thymus, liver, stomach and ileum was followed during 33days post-transplantation. On 8 and 33days post-transplantation, we found massive presence of donor (lacZ(+)) cells in the thymic cortex. Hematopoietic stem cell transplantation led not only to recovery of hematopoietic and lymphoid tissues but also facilitated recovery of the small intestinal mucosa, which was significantly damaged by ionizing radiation.

Introduction: Alcohol and caffeine is intermittently or regularly used by majority of people. These drugs may have acute or chronic effects on patients with traumatic brain injury. Alcohol intoxication increases cerebral blood flow from 8 to 24%. Caffeine decreases cerebral blood flow from 10 to 20%. These facts create a theoretical hypothesis that the decrease of CBF may reduce incranial pressure. The aim of this study was to investigate the effect of caffeine on intracranial pressure in rats following traumatic brain injury., Methods: Ten Wistar male rats underwent weight drop closed head injury. The second stage of the experiment was done 12 to 18 hours later. Intracranial pressure, respiration, heart rate and the mean arterial pressure was monitored. Intraperitoneal injection of caffeine (20 mg/kg) was administered and repetitive measurement of vital signs and intracranial pressure was done., Results: The baseline ICP after head injury was 8.5 +/- 2.9 mm Hg. Ten minutes after intraperitoneal caffeine administration ICP dropped to 7.6 +/- 3.1 mm Hg (p < 0.05). This represents a 11% decrease from baseline value. Mean arterial pressure, respiration and heart rate were stable., Conclusion: Intracranial pressure decrease of 11% from baseline value. This drop was compatible with our hypothesis. The exact dosage of caffeine, its form and rate, should be more precisely specified in further studies.

Introduction: Hemaphereses are sophisticated procedures performed for many indications even in severely ill patients. Many authors consider quite necessary to register as many details as possible of treatments with therapeutic apheresis. WAA meets the requirement to compare data with centers not performing apheresis for the same diagnosis. In Czech Republic hemaphereses are used in a broad spectrum of indications. Since the year 2004 data on hemaphereses done in Czech Republic have been registered. In this paper we present a survey of our to date recordings., Patients and Methods: Data of performed therapeutic hemaphereses (plasma exchange, erythrocytapheresis, leukapheresis, thrombocytapheresis, photopheresis, immunopheresis, and rheopheresis) have been entered in WAA registry with many details. We have been evaluated 1289 procedures in 216 patients done in our two centers (center I, center II)., Results: Center I registered 129 procedures in 41 patients, center II 1260 procedures in 175 patients. The patients are divided according to centers specialization (center II has registered 12 long-term treated patients with LDL-apheresis; mean time of therapy 7.1 years and a median of 34 procedures/patient. Side-effects registered in center I and center II were 3.1% and 5.6% of the procedures, respectively. Most frequent side-effects were citrate toxicity, neurovegetative lability, problems with venous access and hypotension. All were easy to treat, no serious events or death occurred., Conclusions: In Czech Republic hemaphereses are performed in a broad scale of indications and now it is nearly 5 years that data are registered in our two university centers. In 2004 we entered WAA registry because it meets the requirement to compare data with centers that do not perform aphereses for the same diagnosis. This comparison would certainly improve efficacy of the hemapheresis therapy even further. To enter WAA registry is easy, at no expense and without any problems.

Simvastatin and atorvastatin belong to the group of hypolipidemic drugs, more exactly to the second generation of inhibitors of microsomal 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase. They induce a significant reduction in total cholesterol, low-density lipoprotein cholesterol and plasma triglycerides, therefore they are widely used in the treatment of hypercholesterolemia even of its severe form-familiar hypercholesterolemia. Simvastatin and atorvastatin as the most widely used statins in clinical treatment and their hydroxy-acid/lactone forms were determined by means of UPLC in connection with triple quadrupole mass spectrometer. Deuterium labeled reference standard compounds were used as internal standards for the quantitation. Separation was performed on Acquity BEH C18 (100 mm x 2.1 mm, 1.7 microm) using gradient elution by mobile phase containing acetonitrile and ammonium acetate pH 4.0, which is convenient in order to prevent interconversion of analytes. ESI in positive mode was used for the ionization of all compounds. Two SRM (selected reaction monitoring) transitions were carefully optimized for each analyte in order to get high sensitivity and selectivity. SPE on Discovery DSC-18 was used as a sample preparation step. Intra-day precision was generally within 10% RSD, while inter-day precision within 15% RSD. Method accuracy expressed as recovery ranged from 75 to 100%. The method was validated with the sensitivity reaching LOQ 0.08-5.46 nmol/l and LOD 0.01-1.80 nmol/l in biological samples. Atorvastatin, simvastatin, its metabolites and hydroxy-acid/lactone forms were monitored in human serum and in lipoprotein fractions (LDL, HDL and VLDL) at patients with end stage renal diseases.

Objectives: The purpose of this study was to assess whether low serum levels of fetuin-A are potential biochemical predictor of early and/or late survival in chronic hemodialysis (HD) patients., Design and Methods: We measured serum levels of fetuin-A in 67 patients on chronic HD, and correlated it to 3, 12, and 24 months mortality., Results: Cumulative death rate was 7%, 19%, and 37% deaths at 3, 12, and 24 months. Serum fetuin-A was significantly lower in 3 months and 12 months non-survivals (p<0.001), but not in 24 months non-survivals. Kaplan-Meier analyses based on fetuin-A tertiles showed statistically significantly increased probability of death up to 12 months of follow-up for decreasing fetuin-A concentrations (p<0.008)., Conclusions: Fetuin-A as a circulating inhibitor of vascular calcification was significant predictor of early mortality in chronic HD patients but did not appear as a fair marker for later survival.

Introduction: The age related macular degeneration (ARMD) is the most common cause of practical blindness in the population older than 60 years of age. Usually it starts as a dry form with soft drusen.The progression of the dry form is difficult to control-promising seems to be the influence of rheologic factors in the choroid and the retina and thus improvement of their blood flow., Aim: By means of haemorheopheresis to improve blood rheology and thus micocirculation in the choroid and retina. The authors describe the development of the dry form of the ARMD in the sensitive stage with soft drusen and the eventual presence of retinal pigment epithelium detachment (RPED) after haemorheopheresis. The treated patients were compared with a control group (natural course of the ARMD) with the same number of patients., Methods: In a prospective, randomized study, comprising of 16 patients with the dry form of ARMD were treated with haemorheopheresis (cascade filtration) and 16 patients in the control group. Haemorheopheresis treatment comprised of 8 cycles in 10 weeks. Plasma obtained by separator was filtered by the Evaflux 4A (Japan) separator, where, the defined spectrum of high molecular proteins was captured. This lead to a decrease in blood and plasma viscosity and thus to improvement of rheologic conditions of the retina., Results: Results show a positive influence on the absorption of soft drusen, reduction of the retinal pigment epithelium detachment, and the possibility to improve the visual acuity of the treated patients., Conclusion: Comparing to the worsening of clinical signs and visual functions in natural course of the ARMD development, the rheopheresis as well tolerated method may cause the retardation or stop of the unfavorable course of the disease.

Rheological conditions basically influence tissue perfusion, oxygen and nutrient supply, tissue regeneration and in its consequence the course of pathological processes, especially in microcirculation and partially even in macrocirculation. Haemorheotherapy has shown to be successful in several indications (critical disorders in microcirculation) when other methods of therapy have failed. In this study we describe the changes of effectivity indicators in haemorheopheresis treatment and their clinical importance in cases of eye microcirculation, statoacoustic apparatus and disorders of the lower extremities. We treated with haemorheopheresis (separator Cobe.Spetra + Evaflux filter) 24 patients (16 patients with age related macular degeneration, 5 with critical peripheral arterial foot disease, and 3 with acute hearing loss). After the procedures alpha2-macroglobulin decreases by about 58%, fibrinogen by about 69%, IgM by about 61%, LDL-cholesterol by about 77%, apolipoprotein B by about 76% and lipoprotein(a) by about 63%. It corresponds with a decrease in blood and plasma viscosity and clinical improvement in the observed patients, i.e. visual improvement, acceleration of tissue defect healing and improvement in hearing. We noticed 7.1% clinically insignificant side-effects. The method of haemorheopheresis was safe. It led to improvement in patients' clinical condition in the above mentioned indications when other methods of treatment failed.

The aim of this work is to arbitrate the incidence of side effects and tolerability of long lasting LDL-apheresis in familial hyperlipoproteinemia. 1200 procedures were performed and the last 463 of them were evaluated. An immunoadsorption method of LDL-apheresis was used (continuous blood cell separator Cobe Spectra; secondary device: automated adsorption-desorption ADA, Medicap; absorption columns: Lipopak). As a whole, 6.26% adverse events were found and subsequently resolved by standard symptomatic therapy. Vaso-vagal reactions (symptoms of neurovegetative lability) were the most common adverse effects, presented as malaise, weakness, slight and short-term drop in blood pressure or other general signs. They were all well controlled by symptomatic therapy. We conclude that LDL-apheresis in the hands of experienced personnel is a safe procedure. An acceptable procedure duration limit, balancing the possibility to achieve a targeted cholesterol level while still maintaining an acceptable patient tolerance, was confirmed to be 4 hours.

Metabolic abnormalities frequently associated with type 2 diabetes, feature besides endothelial dysfunction a novel factor of low cholesterol absorption and high cholesterol synthesis. We hypothesized an association between endothelial dysfunction and disturbances in cholesterol turnover, predisposing advanced atherosclerosis in patients with type 2 diabetes. We studied 75 patients: 30 with type 2 diabetes, 30 non-diabetic subjects with a history of cardiovascular disease, and 15 healthy subjects. Plasma sterols, soluble adhesion molecules sCD14, sCD40 Ligand, monocyte chemo-attractant protein-1 (MCP-1), sE- and sP-selectins were measured with and without atorvastatin therapy. The diabetic patients showed significantly higher levels of lathosterol and lower levels of sitosterol and campesterol. Non-diabetic subjects showed no significant differences in non-cholesterol based sterols. Plasma levels of hsCRP, sE- and sP-selectins and MCP-1 were significantly increased in patients with diabetes. The plasma levels of sCD40L correlate significantly with clinical parameters of BMI and glycaemia, and the plasma levels of sP-selectin correlate significantly with parameters of glycaemia and HbA(1c). The diabetic patients without statin therapy showed a significant correlation of sE-selectin with the marker of cholesterol absorption-sitosterol and sitosterol/cholesterol ratio. The diabetic patients without statin therapy showed a significant inverse correlation of sP-selectin with the marker of cholesterol synthesis-lathosterol. Both relationships disappeared with statin treatment. We conclude that endothelial dysfunction in obese patients with type 2 diabetes mellitus and cardiovascular disease associates with obesity and glycaemic control. The relation of parameters of endothelial dysfunction (such as sP-selectin and sE-selectin) with cholesterol synthesis and absorption may be influenced by reverse cholesterol transport.

Group of 152 patients (investigated before autologous transplantation) and 35 healthy donors for allogeneic transplantation was examined for the risk of infection transmission that can be associated with the infusion of cryopreserved peripheral blood progenitor cells to the patient and/or cross-contamination of stored grafts. No laboratory signs of active infection were found in 22 donors (63 %) and in 91 patients (60%). The most common was active infection by herpes viruses--50 cases in patients, 21 cases in donors; hepatitis B was found in only two cases. The rate of clinically unsuspected (but dangerous) infections in donors and patients thus remains relatively high in spite of the fact that the system of donor search and the whole transplantation procedure have improved in the last years. The system of safety assurance is extremely important and the whole palette of preventive tests according to EBMT (European Blood and Marrow Transplantation Group) and ISHAGE (International Society for Hemotherapy and Graft Engineering) is fully justified.

Background: Molecular biology methods based on reverse transcription and polymerase chain reaction (RT-PCR) are able to detect the presence of BCR-ABL transcripts in chronic myeloid leukemia (CML). In this study we present our experience with monitoring of residual disease using real-time PCR with hybridization probes detection in patients treated with imatinib mesylate and in collected peripheral blood progenitor cells (PBPC)., Methods and Results: We measured the level of BCR-ABL transcripts in peripheral blood cells of 27 subjects before and in the course of the imatinib treatment. The median of relative quantity of BCR-ABL in the blood before imatinib therapy was 2.55%. The number of the transcripts in 23 imatinib-sensitive subjects decreased to 0.02% in 6 months. After 12 months of the treatment the BCR-ABL median was 0.005%. Subsequent levels fluctuated between values below the detection limit (DL, 0.001%) and 0.005%. Three patients were primarily resistant to imatinib with the BCR-ABL range of 0.13%-11.7% during the treatment. One subject showed marks of molecular relapse after 18 months of the treatment. Only two of 16 filgrastim-stimulated patients had BCR-ABL levels in the blood and in collected PBPC below DL. In other subjects BCR-ABL transcripts were determined within the measurable range of RT-PCR., Conclusions: Taking into account prognostic importance, the measurement of BCR-ABL transcripts is an effective approach to monitoring of residual CML kinetics. Evaluation of BCR-ABL levels in collected PBPC can complete information on quality of the cells in potential autotransplants, and choose subsequent therapeutic protocols and patient prognosis.

A number of surprising observations have shown that stem cells, in suitable conditions, have the ability to produce a whole spectrum of cell types, regardless, whether these tissues are derived from the same germ layer or not. This phenomenon is called stem cell plasticity, which means that tissue-specific stem cells are mutually interchangeable. In our experiments, as a model, we used neural stem cells (NSCs) harvested from fetal (E14-15) neocortex and beta-galactosidase positive. In the first experiment we found that on days 12 and 30 after sub-lethal irradiation (LD 8.5 Gy) and (beta-galactosidase(+)) NSCs transplantation all mice survived, just as the group with bone marrow transplantation. Moreover, the bone marrow of mice transplanted NSCs contained the number of CFU-GM colonies with beta-galactosidase(+) cells which was as much as 50% higher. These differences were statistically significant, p<0.001. In the second experiment, we studied kinetics of (beta-galactosidase(+)) NSCs after their transplantation to sub-lethally irradiated mice. Histochemistry of tissues was performed on days 12 and 30 post-transplantation, and beta-galactosidase(+) cells were detected with the help of histochemical examination of removed tissues (lung, liver, spleen, thymus, and skeletal muscle). In tissues removed on day 12 post-transplantation, we found a significantly higher number of beta-galactosidase(+) cells in the spleen and thymus on day 30. While we presumed the presence beta-galactosidase(+) cells in the spleen, as spleen and reticuloendothelial system represent an important retaining system for different cell types, the presence of beta-galactosidase(+) cells in the thymus was rather surprising but very interesting. This indicates a certain mutual and close interconnection of transplanted stem cells and immune system in an adult organism. In the third experiment, we verified the mutual interchange of Sca-1 surface antigen in the bone marrow cells and NSCs before transplantation. Analysis of this antigen showed 24.8% Sca-1 positive cells among the bone marrow cells, while NSCs were Sca-1 negative. Our experiments show that NSCs share hemopoietic identity and may significantly influence the recovery of damaged hematopoiesis but do not have typical superficial markers as HSCs. This result is important for the determination of predictive factors for hemopoiesis recovery, for stem cell plasticity and for their use in the cell therapy.

Background: The interest in aspirin resistance has been increasing during the last few years, with researchers earnestly pursuing alternative anti-platelet therapies and devices for measuring platelet aggregation. The recent studies suggest that 5-45% of patients taking aspirin do not experience adequate anti-platelet effects., Methods and Results: There is scant evidence proving that aspirin resistance has some clinical consequences. To assess the prevalence of aspirin resistance in patients with ischemic heart disease (IHD) two independent methods were used: platelet aggregation in platelet rich plasma (PRP) after induction by propylgallate (CPG), and assessment of platelet function by PFA 100 method. The study population consisted of 424 patients treated for IHD on the 2nd Dept. of Medicine, Teaching Hospital, Hradec Kralove. The age, gender, diagnosis and effect of therapy were characterized in this group of the patients. Daily ASA dose was 100 mg. We used two methods to monitor ASA treatment efficacy: a) thrombocyte aggregation after induction by CPG, b) the assessment of platelet function by PFA 100 method. a) Of the patients studied by CPG platelet aggregation, 51 (12.1%) pts were resistant to ASA dose 100 mg/day, and 32 (7.6%) pts remained resistant even after increasig the dose to 200 mg/day. In 80 (20%) pts, the daily ASA dose of less than 100 mg was sufficient to inhibit platelet function. b) Although the PFA-100 system is not able to detect the difference between low and high ASA dose, we found 53 (15.2%) patients aspirin resistant using this method., Conclusions: In the patients with IHD treated with 100 mg of ASA per day, our study has shown that the prevalence of aspirin resistance was 12.1% using CPG method, and 15.2% using PFA-100 method. Aspirin resistance was dose dependent. Prevalence of ASA resistance in patients treated with 200 mg of ASA per day was only 7.6% using the CPG method.

The surgical anatomy of the temporal branch of the facial nerve was studied in the anatomical laboratory. The temporal branch divides into an anterior, middle (frontal), and a posterior ramus after it pierces the parotid fascia. The anterior ramus innervates orbicularis oculi and corrugator supercilii muscles; the middle branch is for the ipsilateral frontalis muscle. The posterior branch innervates the anterior and superior auricular and tragus muscles. Below the zygomatic arch, the temporal branch of the facial nerve is located in the subcutaneous tissue. Above the arch, it continues in the subgaleal space with the superficial temporal fascia deeply. The terminal twigs of the temporal branch penetrate the galea to reach their target muscles that are all located superficial to the galea. There is a significant variability in the course of the temporal branch of the facial nerve. Occasionally, the terminal twigs of the middle ramus may penetrate superficial layer of superficial temporal fascia and run in the intrafascial fat pad before entering the frontalis muscle. There are four available operative techniques in this anatomical location. The subgaleal dissection of a temporofrontal scalp flap is associated with a high incidence of postoperative palsy of the temporal branch of the facial nerve and cosmetically bothersome results. Reflecting the scalp and temporalis muscle together as a single layer is the safest procedure. Unfortunately, this technique can not be used for the transzygomatic approaches and the bulky temporalis muscle may compromise basal exposure in the pterional route. Third technique was described and propagated by Yasargil. He proposed a subgaleal dissection up to the anterior one-fourth of the temporalis muscle where the dissection has to be deepened between the two layers of the superficial temporal fascia (in the interfascial fat pad). This approach may also infrequently injure the temporal branch in case of anatomical variation. The last available operative technique raises the superficial temporal fascia together with the scalp.

The role of adhesive selectin molecules in the process of atherogenesis is an open question. These molecules are known as markers of atherosclerosis activity, however, only some biological mechanisms are known up to now. In this study we examined the levels of soluble forms of E-, P-selectin and monocyte chemotactic protein (MCP-1) in the process of extracorporeal cholesterol elimination by LDL-apheresis. We measured the levels of sE-, sP-selectin and MCP-1 in the plasma before and after LDL-apheresis and in the washout solution from immunoabsorption columns Lipopak. Eighty measurements were performed repeatedly in 6 patients with severe familial hypercholesterolemia (FH) on long-term LDL-apheresis treatment. Before the procedure P-selectin levels were 204+/-179 ng/ml, E-selectin 32.1+/-33.7 ng/ml, MCP-1 323.8+/-121 pg/l, whereas after the procedure we found P-selectin levels 131.6+/-34 ng/ml, E-selectin 33.1+/-51 ng/ml, and MCP-1 200.4+/-15 pg/l. Levels of P-selectin were increased in the blood of patients with FH in spite of long-term intensive extracorporeal LDL-elimination, documenting thus the activity of atherosclerosis. The levels of P-selectin and MCP-1 decreased significantly after the hypolidemic procedure and could be used as another marker showing the effectivity of the extracorporeal LDL-cholesterol elimination (immediately after the procedure), and, after further verification, may serve as a marker for controlling the therapy efficacy.

We report here the case report of patient with empyema of the thorax and sternal osteomyelitis, which undertool removal of ectopic mediastinal parathyroid adenoma with omentoplasty of sternal defect. Osteomyelitis was cured; though empyema persisted for several months and it's healing was complicated with central venous catheter sepsis.