[MicroRNA Analysis for Extramedullary Multiple Myeloma Relapse]. / Analýza mikroRNA u extramedulárního relapsu mnohocetného myelomu.

INTRODUCTION AND AIMS: Multiple myeloma (MM) is the second most common hematooncological disease. Patient survival has been greatly improved by the introduction of new drugs into clinical practice, but survival is negatively affected by the so-called extramedullary relapse (EM), caused by the loss of plasma cell dependence on the bone marrow microenvironment and their migration out of the bone marrow. The nature and causes of this process are currently unclear. MicroRNAs (miRNAs) are short, non-coding RNA molecules involved in many physiological and pathological processes. Their significance in the pathogenesis of MM has been demonstrated by several studies. We assume that they are also involved in the development of the EM. The aim of this study was to analyze different miRNA expression between MM and EM patients. MATERIAL AND METHODS: Using next generation sequencing, we analyzed 39 samples of bone marrow cells from MM patients at diagnosis and 9 bone marrow plasma samples of EM patients. RESULTS: In total, 2,278 miRNA were sequenced, but only 658 miRNAs were analyzed as they were expressed in all samples and had at least 20 reads. Expression data were generated using the Chimira tool from fastq data. All sequences were mapped using miRBase v20. Further analyses were performed using the R/Bioconductor package. The Bayesian procedure was used for normalization of expression. P values were adjusted using the Benjamini-Hochberg method. Analysis found 10 miRNA (p < 0.0005) that are statistically significantly expressed in EM vs. MM patients - these are miR-26a-5p, miR-26b-5p, miR-30e-5p, miR-424-3p, miR-503-5p, miR-767-5p, miR-105-5p, miR-5695-5p, miR-450b-5p and miR-92b-3p. These miRNAs will be further verified by qPCR method on a larger set of MM and EM patients. CONCLUSION: Our pilot study has shown that there are differentially expressed miRNAs between MM and EM patients.Key words: multiple myeloma - microRNA - carcinogenesis - next generation sequencing The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papersThis work was supported by grant MZ CR AZV 17- 29343A. Submitted: 17. 3. 2018Accepted: 20. 3. 2018.

[Urinary MicroRNAs as Potential Biomarkers of Bladder Cancer]. / Mocové mikroRNA jako potenciální biomarkery karcinomu mocového mechýre.

BACKGROUND: Currently, there are no urinary-based tumour markers with sufficient sensitivity and specificity to replace cystoscopy in the detection of bladder cancer (BCA). Urinary microRNAs are emerging as clinically useful class of biomarkers for early and non-invasive detection of urologic malignancies. PATIENTS AND METHODS: In this study, 155 patients with BCA and 83 healthy controls were enrolled. Expression profiles of urinary miRNAs were obtained using Affymetrix miRNA microarrays and candidate miRNAs further validated in independent cohort using specific TaqMan assays and quantitative real-time polymerase chain reaction method. RESULTS: Whole-genome profiling identified miRNA signature with significantly different concentrations in urine of BCA compared to controls (p < 0.01). In the independent validation phase of the study, three miRNAs were confirmed to have significantly higher levels in urine of patients with BCA in comparison with control groups (p < 0.0001). In addition, we observed significant decrease in two miRNAs (p < 0.01) concentrations in the urinary samples collected 3 months after surgery compared to pre-operative samples. CONCLUSION: We identified and validated miRNAs to have significantly higher concentrations in urine of patients with BCA in comparison with controls. Our data have shown that urinary miRNAs could serve as sensitive and specific biomarkers enabling non-invasive detection of BCA.Key words: urinary microRNAs - biomarkers - bladder cancer The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. This study was supported by Ministry of Health of the Czech Republic, grant No. 15-31071A. All rights reserved.Submitted: 19. 3. 2018Accepted: 20. 3. 2018.

Low-burden TP53 mutations in chronic phase of myeloproliferative neoplasms: association with age, hydroxyurea administration, disease type and JAK2 mutational status.

The multistep process of TP53 mutation expansion during myeloproliferative neoplasm (MPN) transformation into acute myeloid leukemia (AML) has been documented retrospectively. It is currently unknown how common TP53 mutations with low variant allele frequency (VAF) are, whether they are linked to hydroxyurea (HU) cytoreduction, and what disease progression risk they carry. Using ultra-deep next-generation sequencing, we examined 254 MPN patients treated with HU, interferon alpha-2a or anagrelide and 85 untreated patients. We found TP53 mutations in 50 cases (0.2-16.3% VAF), regardless of disease subtype, driver gene status and cytoreduction. Both therapy and TP53 mutations were strongly associated with older age. Over-time analysis showed that the mutations may be undetectable at diagnosis and slowly increase during disease course. Although three patients with TP53 mutations progressed to TP53-mutated or TP53-wild-type AML, we did not observe a significant age-independent impact on overall survival during the follow-up. Further, we showed that complete p53 inactivation alone led to neither blast transformation nor HU resistance. Altogether, we revealed patient's age as the strongest factor affecting low-burden TP53 mutation incidence in MPN and found no significant age-independent association between TP53 mutations and hydroxyurea. Mutations may persist at low levels for years without an immediate risk of progression.

Prognostic significance of miR-23b in combination with P-gp, MRP and LRP/MVP expression in non-small cell lung cancer.

Recently, miR-23b has emerged as a promising new cancer biomarker but its role in lung cancer has not been established yet. Patients still do not respond well to available treatments, probably due to expression of multidrug resistance (MDR) proteins, such as P-gp, MRP and LRP/MVP. The aim of this study was to determine the role of miR-23b in non-small cell lung cancer (NSCLC) and its relationship to the patient outcome together with MDR transporter proteins. We immunohistochemically evaluated expression of P-gp, MRP and LRP/MVP and quantified the relative levels of miR-23b in 62 NSCLC patients´ samples. The prognostic significance of miR-23b and MDR proteins was tested by Kaplan-Meier and Cox-regression analysis. Our results showed that miR-23b is mostly downregulated in NSCLC samples (57/62) and that its upregulation in tumors is connected with longer progression-free survival (PFS; P = 0.065) and overall survival (OS; P = 0.048). The Cox proportional hazard model revealed that the risk of death or relapse in NSCLC patients with miR-23b downregulation increases together with LRP/MVP expression and both risks decrease with miR-23b upregulation (HRPFS = 4.342, PPFS = 0.022; HROS = 4.408, POS = 0.015). Our findings indicate that miR-23b, especially in combination with LRP/MVP expression, might serve as a suitable prognostic biomarker for NSCLC patients.

Detailed analysis of therapy-driven clonal evolution of TP53 mutations in chronic lymphocytic leukemia.

In chronic lymphocytic leukemia (CLL), the worst prognosis is associated with TP53 defects with the affected patients being potentially directed to alternative treatment. Therapy administration was shown to drive the selection of new TP53 mutations in CLL. Using ultra-deep next-generation sequencing (NGS), we performed a detailed analysis of TP53 mutations' clonal evolution. We retrospectively analyzed samples that were assessed as TP53-wild-type (wt) by FASAY from 20 patients with a new TP53 mutation detected in relapse and 40 patients remaining TP53-wt in relapse. Minor TP53-mutated subclones were disclosed in 18/20 patients experiencing later mutation selection, while only one minor-clone mutation was observed in those patients remaining TP53-wt (n=40). We documented that (i) minor TP53 mutations may be present before therapy and may occur in any relapse; (ii) the majority of TP53-mutated minor clones expand to dominant clone under the selective pressure of chemotherapy, while persistence of minor-clone mutations is rare; (iii) multiple minor-clone TP53 mutations are common and may simultaneously expand. In conclusion, patients with minor-clone TP53 mutations carry a high risk of mutation selection by therapy. Deep sequencing can shift TP53 mutation identification to a period before therapy administration, which might be of particular importance for clinical trials.

Diffuse large B-cell lymphoma: the history, current view and new perspectives.

The basic principles of lymphoma classification(s) in general have been widely evolving in a course of decades of years wiht the use of contemporary resources and recent cutting edges in hematooncology on a clinical, morphological and molecular level bring new possibilities not only in improvements of diagnostic and prognostic algorithms and also bear new opportunities in so called targeted and tailored strategies of lymphoma therapy. The pathogenesis and biologic behavior of lymphoproliferations and even lymphomas should be studied in a context of lymphocytic and (neoplastic) lymphoid stage and chronologic development. In a current more complex insight into lymphoproliferations we would like to describe huge heterogeneity of diffuse large B-cell lymphoma in relationship to mandatory WHO classification since 2008 and the next development of knowledge in this field with potential new influence on an advancement of both classification and therapy.

The incidence of metabolic syndrome in obese Czech children: the importance of early detection of insulin resistance using homeostatic indexes HOMA-IR and QUICKI.

Common alimentary obesity frequently occurs on a polygenic basis as a typical lifestyle disorder in the developed countries. It is associated with characteristic complex metabolic changes, which are the cornerstones for future metabolic syndrome development. The aims of our study were 1) to determine the incidence of metabolic syndrome (based on the diagnostic criteria defined by the International Diabetes Federation for children and adolescents) in Czech obese children, 2) to evaluate the incidence of insulin resistance according to HOMA-IR and QUICKI homeostatic indexes in obese children with and without metabolic syndrome, and 3) to consider the diagnostic value of these indexes for the early detection of metabolic syndrome in obese children. We therefore performed anthropometric and laboratory examinations to determine the incidence of metabolic syndrome and insulin resistance in the group of 274 children with obesity (128 boys and 146 girls) aged 9-17 years. Metabolic syndrome was found in 102 subjects (37 %). On the other hand, the presence of insulin resistance according to QUICKI <0.357 was identified in 86 % and according to HOMA-IR >3.16 in 53 % of obese subjects. This HOMA-IR limit was exceeded by 70 % children in the MS(+) group, but only by 43 % children in the MS(-) group (p<0.0001). However, a relatively high incidence of insulin resistance in obese children without metabolic syndrome raises a question whether the existing diagnostic criteria do not falsely exclude some cases of metabolic syndrome. On the basis of our results we suggest to pay a preventive attention also to obese children with insulin resistance even if they do not fulfill the actual diagnostic criteria for metabolic syndrome.

[Evolution of the technique of arthroscopic reinsertion of the rotator cuff. Our experience from the years 1998 to 2008]. / Vývoj operacní techniky artroskopické reinzerce manzety rotátoru: nase zkusenosti z let 1998-2008.

PURPOSE OF THE STUDY: A rotator cuff tear is a relatively frequent cause of pain and restricted motion of the shoulder. Some orthopaedists believe that any attempt at rotator cuff reconstruction will fail. The aim of this paper is to present our experience with arthroscopic reconstruction of rotator cuff tears. MATERIAL AND METHODS: Between January 1998 and December 2008, 319 patients with an early diagnosis of rotator cuff rupture were treated. The group included 67 women and 252 men, with an average age of 37 years (range, 24 to 71 years) at the time of surgery. The patients indicated for arthroscopic reconstruction had to show free motion of the shoulder, had a full thickness tear up to 3 cm in size in the sagittal plane and a Patte stage 2 tear in the frontal plane at the maximum. The outcome of surgery was evaluated at one year of follow-up and included the patient's self-assessment, modified UCLA score and incidence of complications. The probability of failure was calculated as an odds ratio of an implant failure to failure of the other implants and the probability of repeat surgery in a given implant was calculated as a relative risk in relation to the other implants. RESULTS: The average operative time was 52 minutes (range, 25 to 85); the average UCLA score increased from 10 to 31 points (p<0.00001). An excellent or a good result was achieved in 80% of the patients. Rotator cuff reconstruction failed in 32 patients (11%), of whom 22 (7.6%) underwent revision surgery. The failure was due to migration of rotator cuff anchors or thread failure in 14 patients (14/32; 44%). The GII anchors showed the highest risk of failure, with the odds ratio of 5.55 (95 % CI, 2.22 to 13.84) for mechanical failure of the method and a relative risk of revision surgery of 7.62 (95% CI, 2.86 to 20.27). For comparison, the RC anchors had the odds ratio for mechanical failure equal to 0.55 (95 % CI, 0.25 to 1.24) and the relative risk of repeat surgery equal to 0.41 (95% CI, 0.12 to 1.43). In addition, 18 complications were recorded. The frequency of deep wound infection was 0.7% (2/319). Six patients (2.1%) required repeat surgery for symptomatic bursitis and adhesive capsulitis. DISCUSSION: A recent meta-analysis has found no significant difference between the results of surgical rotator cuff reconstruction and its conservative treatment. We do not support this view but present here evidence that, when certain conditions are fulfilled, arthroscopic reconstruction can produce a very good clinical outcome. CONCLUSIONS: The arthroscopic reconstruction of a rotator cuff tears results in a marked relief of pain and improved joint function. An ideal candidate for this treatment should show passive free motion at the shoulder joint, no clinical signs of bursitis, and mobilisable tendon stumps of the torn rotator cuff. In addition, these patients should be highly motivated for post-operative rehabilitation. A suture device was most effective in rotator cuff repair. For good fixation into the bone it is recommended to use special implants that have a minimal risk of dislodgement or anchor thread failure.

[Cementless Plasmacup-Bicontact total hip arthroplasty. Results of a minimum of ten-year follow-up]. / TEP kycle Plasmacup-Bicontact: výsledky s minimálne 10letým sledováním.

PURPOSE OF THE STUDY: The aim of the study was to analyse the long-term clinical and radiographic results of total hip arthroplasty (THA) with the Bicontact-Plasmacup prosthesis, MATERIAL AND METHODS: In this retrospective observational study, 34 consecutive patients undergoing Bicontact-Plasmacup THA between August 1998 and July 2000 were evaluated. The group included 15 women and 19 men with an average age of 56 years (30 to 67 years). The mean ± SD of follow-up was 132 ± 9 months (122 to 149 months). Patients' satisfaction with the treatment outcome and selected clinical and radiographic features were evaluated. The data were analysed using the concordance test, Spearman's correlation coefficient and chi-square test. RESULTS: At the final follow-up, 94% of the interviewed patients reported satisfaction with the surgery outcome. The mean Harris score was 93 (67 to 98) points, with excellent or very good results in 91% of the patients. Compared with the post-operative radiographs, the mean ± SD value for vertical migration of the acetabular cup was 2.43 ± 3.21 mm (0 to 12 mm) and that for horizontal migration was 0.35 ± 1.0 mm (0 to 4.3 mm). The mean stem subsidence was by 3.87 mm (range, 0.36 to 21.11 mm; SD 4.03). At the final follow-up, all implants showed radiographic stability and absence of radiolucent lines. Early acetabular osteolysis was detected in two patients (5.9 %). In 19 patients (56%) a change in bone architecture, similar to alterations due to weight-bearing adaptation, was seen around the cup following surgery. The stress shielding effect of the femoral component was recorded in 79%, bypass phenomenon in 41 % and pedestal formation under the apex of the stem in 18% of the patients. The mean rate of polyethylene wear measured by the Dorr and Wan method was 0.106 mm per year (range, 0.00 to 0.267 mm) or it was 0.078 mm per year (0.00 to 0.19 mm) when assessed by a modification of the method described by Kang et al. DISCUSSION: The clinical and radiographic findings presented here are in accordance with the relevant literature data. This implant design is associated with a relatively infrequent osteolysis around the cup probably due to a low wear rate of polyethylene and a stable polyethylene liner-metal cup interface. Based on this work and some other studies it can be concluded that a post-operative migration of several millimetres is compatible with a long-term, stable cementless THA. CONCLUSIONS: The Bicontact-Plasmacup THA meets the National Institute for Health and Clinical Excellence (NICE) criteria for a very good implant (ten-year survival rate of over 90%). At 10 years after surgery, however, the first signs of complaints about surgery outcome // dissatisfaction with surgery outcomes were recorded. This may, at least partly, be related to aseptic loosening which is one of the symptoms of particle disease.

[Experiences and first outcome of posterior ankle arthroscopy]. / Artroskopie zadního hlezna - nase zkusenosti a první výsledky.

PURPOSE OF THE STUDY: Many hind foot problems can be treated by endoscopic procedures. It is essential to identify the best candidates for this treatment and also to assess the risk associated with these surgeries. Here we present our first experience with posterior ankle arthroscopy. MATERIAL AND METHODS: Between September 2007 and September 2009, we performed 21 arthroscopic procedures on the posterior ankle (one-stage combined anterior and posterior surgery, 11x; posterior procedure, 7x; two-stage anterior and posterior surgery, 3x). The group included 21 patients, 18 men and three women, with an average age of 36 years (20 to 64) at the time of surgery. The average follow-up was 27 months (16 to 38). Both posterolateral and posteromedial approaches were used, with major landmarks being the processus posterior tali, posterior talofibular ligament and long flexor tendon of the big toe. Surgery was performed by experienced specialists. Outcome evaluation was based on the AOFAS and Kitaoka clinical rating scales and the complication rate. RESULTS: In our group, the diagnoses treated by arthroscopic surgery were as follows: arthrofibrosis (16x; 76 %), hypertrophic processus posterior tali (13x; 62 %), and loose intra-articular bodies (8x; 38 %). All patients reported improvement in clinical condition after surgery. The differences between the mean pre- and post-operative values were 24 points for the AOFAS score (58 versus 82; p<0.00015) and 23 points for the Kitaoka score (58 versus 81; p < 0.00015). The most effective outcomes were achieved in the patients operated on for loose bodies or hypertrophic processus posterior tali. The worst result was found in the patient with advanced arthritis of the ankle that had to undergo arthrodesis 13 months following arthroscopic treatment. There were eight complications in five patients (24 %), including excessive bleeding in five cases, temporary loss of sensation in two, and a damaged tendon of the long flexor of the big toe. DISCUSSION: Arthroscopic surgery has recently been advocated by many authors. The majority of them have reported good results and an acceptable risk of complications. Almost all use either the posteromedial or the lateral approach, which are anatomically safe procedures. Surgery is followed by a thorough rehabilitation programme that allows most patients to resume their normal daily activities in 8 weeks. Our results were not so good because we mostly treated post-traumatic conditions, with advanced ankle arthritis in several patients. CONCLUSIONS: Arthroscopy is an effective method for treating both intra- and extra-articular pathologies in the posterior ankle. The best effect of treatment was recorded in posterior tibiotalar impingement syndrome or after extraction of loose intra-articular bodies. In advanced ankle arthritis, on the other hand, the use of arthroscopic treatment should be carefully considered. The risk of complications is high and the procedure requires good knowledge of anatomy and high surgical skills.

[Synovial fluid from aseptically failed total hip or knee arthroplasty is not toxic to osteoblasts]. / Výpotky asepticky selhávajících TEP kyclí a kolen nejsou toxické pro osteoblasty.

PURPOSE OF THE STUDY: A failure of total hip or knee artroplasty is associated with an increased production of joint fluid. This contains wear particles and host cells and proteins, and is assumed to be involved in the pathogenesis of aseptic loosening and periprosthetic osteolysis. This study investigated the effect of synovial fluid from patients with aseptically failed joint prostheses on osteoblast cultures. MATERIAL AND METHODS: Synovial fluid samples were obtained from patients with failed total joint prostheses (TJP; n=36) and from control patient groups (n = 16) involving cases without TJP and osteoarthritis, without TJP but with osteoarthritis, and with stable TJP. The samples were treated in the standard manner and then cultured with the SaOS-2 cell line which shows the characteristics and behaviour of osteoblasts. Each fluid sample was also examined for the content of proteins, cells and selected cytokines (IL-1ß, TNF-α, IL-6, RANKL and OPG detected by ELISA). We tested the hypothesis assuming that the fluids from failed joints would show higher cytotoxicity to osteoblast culture and we also expected higher levels of IL-1ß, TNF-α, IL-6, and RANKL in patients with TJP failure and/ or with more severe bone loss. The statistical methods used included the Kruskal-Wallis ANOVA and Mann-Whitney U test. RESULTS: The fluids from failed TJPs showed the highest RANKL and the lowest OPG levels resulting in the highest RANKL/OPG ratio. However, there was no evidence suggesting that the joint fluids from failed TJPs would be more toxic to osteoblast culture than the fluids from control groups. In addition, no correlation was found between the fluid levels of molecules promoting inflammation and osteoclastic activity and the extent of bone loss in the hip (in terms of Saleh's classification) or the knee (AORI classification). In fact, the fluids from failed TJPs had higher protein levels in comparison with the controls, but the difference was not significant. DISCUSSION: The finding of high RANKL levels and low OPG concentrations is in agreement with the theory of aseptic loosening and periprosthetic osteolysis. The other cytokines, particularly TNF-α and IL-1ß, were found in low levels. This can be explained by the stage of particle disease at which the samples were taken for ELISA analysis. It is probable that the level of signal molecules reflects osteolytic process activity and is therefore not constant. The reason for no correlation found between cytokine levels and the extent of bone loss may also lie in the use of therapeutic classifications of bone defects that is apparently less sensitive to the biological activity of aseptic loosening and/or periprosthetic osteolysis. CONCLUSIONS: Synovial fluids from failed total hip or knee joint prostheses are not toxic to osteoblast cultures. Cytotoxicity indicators and levels of pro-inflammatory and pro-osteoclastic cytokines (IL-1ß, TNF-α, IL-6, RANKL and OPG) do not correlate well with the extent of periprosthetic bone loss. Key words: total joint replacement, arthroplasty, aseptic loosening, periprosthetic osteolysis, joint fluid, SaOS-2 cell line, cytotoxicity, cytokines, RANKL, OPG.

[Detection of minimal residual disease and its significance for establishing prognoses in patients with laparoscopic resections of colorectal carcinomas]. / Detekcia minimálnej reziduálnej choroby a jej význam pre prognózu pacientov s laparoskopickou resekciou kolorektálneho karcinómu.

AIM: The aim of the study was to detect minimal residual disease (MRD) in bone marrow samples, portal and peripheral blood samples collected from colorectal carcinoma (CRC) patients, and to assess the results in relation with clinical stages of the disorder and to evaluate potential correlation between the MRD presence and the disease relapse and overall patient survival rates. MATERIAL AND METHODS: The study included patients with primary CRCs indicated for laparoscopic resections. From September 21, 2006 to December 31, 2008, the authors selected 159 subjects with median age of 56. 126 patients (79.25%) were operated for CRC stage I-III, 33 patients (20.75%) had CRC stage IV. Six samples were collected in each subject to detect the MRD presence (systemic venous blood and bone marrow at the beginning of the procedure, venous blood from the mesenteric bed, systemic venous blood after the resection procedure, systemic venous blood and bone marrow one month after the procedure), as well as samples of the tumor tissue. Real-time RT-PCR method was use to detect the MRD. RESULTS: The study confirmed correlation between MRD positivity in preoperative bone marrow samples and the disease stage (p < 0.035). It showed correlation between findings in preoperatively collected systemic venous blood samples and in mesenteric venous blood samples (p < 0.003), correlation between findings in systemic venous blood samples collected after the resections procedures and in systemic venous blood samples one month after the procedure (p < 0.015), as well as correlation between findings in preoperative systemic venous blood samples and findings in systemic venous blood samples collected after the procedures (p < 10(-5)). The authors found out that the surgical procedure affected the MRD presence in systemic venous blood samples in primary negative patients (p < 0.025). During the study period, the authors revealed no statistically significant correlation between the MRD findings in stage I-III patients and their disease-free survival (p < 0.59). Considering the above results, possible direct correlation between positive MRD findings in systemic venous blood samples, which were collected preoperatively in CRC stage I-III patients, and reduced survival time is expected (p < 0.075). During the study period, the overall survival time was significantly reduced in stage I-III patients with positive findings in postoperative systemic venous blood samples, compared to that in negative patients (p < 0.004). CONCLUSION: The data suggest certain correlations between the MRD findings and the disease prognosis. The authors continue to collect further samples and assess the outcomes in order to enlarge the patient study group and the data, and plan to evaluate the outcomes in a 5-year to 10- year follow up period.

[Minimal residual disease in pancreatic cancer--our first experiences]. / Minimální reziduální choroba u karcinomu pankreatu--nase prvotní zkusenosti.

INTRODUCTION: Pancreatic cancer is one of the most aggressive malignity with the statistically shown an upward trend and a very poor prognosis. The causes we follow up in the local recurrence and in the early dissemination, either through hematogenic or lymphogenic way. Conventional methods are not able to capture these cells, just the modern molecular-biological methods make possible to fix the so-called minimal residual disease, that is the presence of isolated tumor cells in the patient's body. MATERIAL AND METHODS: The study included 52 patients operated on the Clinic of Surgery I. in the University Hospital Olomouc for pancreatic cancer in different stages. QRT-PCR method was determined expression of hTERT, EGFR 1 and CEA both in peripheral blood, portal blood, bone marrow, peritoneal lavage and the tumor itself. RESULTS AND CONCLUSION: The results of this pilot study demonstrated a high sensitivity and specificity of the PCR method for detection of circulating tumor cells in patients with pancreatic cancer, extending this methodology, we are able to provide prognostic value of minimal residual disease and its significance for the indication of radical surgery for pancreatic cancer.

Increased lipid peroxidation and abnormal fatty acid profiles in seminal and blood plasma of normozoospermic males from infertile couples.

BACKGROUND: Reactive oxygen species (ROS) are essential for sperm function. However, excessive ROS production can impair sperm function and might be a factor contributing to male infertility. METHODS: We investigated the levels of arachidonic acid (AA) and docosahexaenoic acid (DHA) as well as lipid peroxidation, as represented by thiobarbituric acid reactive species (TBARS), in blood and seminal plasma of 38 normozoospermic males from infertile couples (NSI-males), compared with that of 17 fertile volunteers (FV-males). RESULTS: TBARS levels in blood and seminal plasma were higher in NSI-males than in FV-males (P < 0.0002, P < 0.0003, respectively), as were AA levels (P < 0.0003, P < 0.00004, respectively). On the contrary, the blood and seminal plasma levels of DHA were lower in NSI-males than in FV-males (P < 0.02 and P < 0.05, respectively). The AA/DHA ratios in blood and seminal plasma were higher in NSI-males than in FV-males (P < 0.003, P < 0.0007, respectively). Significant correlations between seminal and blood plasma levels of TBARS (P < 0.0001, r = 0.548), AA (P < 0.0001, r = 0.571) and DHA (P < 0.0001, r = 0.506) were found. CONCLUSIONS: Our data provide new insight into lipid metabolism in male infertility and indicate that systemic oxidative stress resulting in increased lipid peroxidation and an altered fatty acid profile may be, at least in part, responsible for infertility even in normozoospermic males.

[Comparison of therapeutic strategies for hip and knee prosthetic joint infection]. / Porovnání lécebných postupu pouzívaných v terapii infekce kloubních náhrad kycle a kolena.

PURPOSE OF THE STUDY: Prosthetic joint infection (PJI) is a feared complication of total joint arthroplasty. Several strategies have been developed to treat it. The purpose of our study was to compare therapeutic strategies for PJI treatment published in the literature. MATERIAL AND METHODS: We retrieved around 5,000 documents concerning this topic, published between January 1960 and November 2006, from the databases MEDLINE and EMBASE. Using a two-phase selection, 382 relevant articles were chosen, and full texts were obtained for 302 of them (79%). However, only 77 of them fulfilled the criteria for inclusion in our study. A total of 645 hips and 1,145 knees could be analyzed. A comparison was made of the following procedures: two-stage surgery, one- stage surgery, implant removal and long-term antibiotic therapy in hip PJI; two-stage surgery, debridement, arthrodesis and long-term antibiotic therapy in knee PJI. The criteria used to evaluate successful PJI treatment included the rate of recur- rent infection (primary outcome) and the rate of additional surgery (secondary outcome). The capability index (c), relative risk (RR), odds ratio and "number needed to treat" (NNT) were calculated for both outcomes under study. RESULTS: 1. The lowest rate of recurrent PJI was reported for two-stage reimplantation (hips, 7.4%; knees, 11%), then came one-stage reimplantation for hip PJI (9.2%) and arthrodesis for knee PJI (15.6%). 2. The lowest relative risk for the primary outcome of this study was found in two-stage reimplantation (RR=0.62 and 0.32 for hip and knee PJI, respectively, knee arthrodesis (RR=0.78) and one-stage hip reimplantation (RR=1.07). 3. The NNT was negative for two-stage reimplantation (NNT = -21.6 and -4.3 for hip and knee PJI, respectively). 4. Compared to long-term antibiotic therapy, knee debridement had a higher risk of recurrent infection (RR, 4.72 versus 1.20) as well as the risk of additional surgery (RR, 4.41 versus 1.31). However, when the capability index and NNT were used, knee debridement achieved better primary outcomes than antibiotic therapy (c=0.0317 versus 0.0000) and NNT (2.2 versus 26.3). 5. Relative risk for additional surgery was the lowest in two-stage reimplantation (RR=0.47 and 0.36 for hip and knee PJI, respectively), and the highest in long-term antibiotic therapy for hip PJI (RR=6.47). DISCUSSION: The resultant ranking of therapeutic approaches to PJI, as found in our study, is not surprising; except for seemingly bet- ter outcomes achieved by long-term antibiotic therapy compared to surgical debridement in knee PJI. This, among others, shows the need to use more comprehensible statistical instruments, such as the capability index, that could provide a more reliable evaluation of therapeutic interventions. CONCLUSIONS: A two-stage protocol for PJI treatment had the lowest risk for both PJI recurrence and need for additional surgery in comparison with all other strategies developed for PJI therapy. Therefore, it should be accepted as the method of choice. One-stage hip reimplantation is a less reliable approach in which it is inevitable to strictly respect the indication criteria. Other methods are either less reliable or associated with a high risk of repeat surgeries, or provide functionally unacceptable outcomes.

[Comparison of reactive oxygen species production in neat semen and washed spermatozoa]. / Porovnání produkce reaktivních kyslíkových cástic v nativním ejakulátu a v suspenzi spermií.

OBJECTIVE: To determine Reactive Oxygen Species (ROS) production in neat semen and spermatozoa suspension using chemiluminescence and to examine correlation between both methods. SUBJECT: Prospective laboratory study. SETTING: Department of Obstetric and Gynecology, University Hospital, Olomouc. METHODS: The study included fertile volunteers (FV, n = 17), men from infertile couples (NM, n = 19) and men with idiopathic infertility (NMI, n = 15). ROS levels were determined by the same method in neat and washed semen samples. RESULTS: The ROS production in neat semen was lower than that in spermatozoa suspension. There was no significant diference in ROS production between volunteers and males from infertile couples. There was a significant correlation between log ROS in neat semen and in spermatozoa suspension in studied groups (FV r = 0.85, p = 1.5 x 10(-5); NM r = 0.76, p < 2 x 10(-4); NMI r = 0.75, p < 1.5 x 10(-3)). CONCLUSIONS: Measurement of ROS in neat semen is simpler, faster and better reflecting the actual level of oxidative stress than the same measurement in spermatozoa suspension. The implementation of this method can complement the algorithm of diagnostics and treatment of male infertility and be helpful in selection of patients for antioxidant or antibiotic treatment.

[Study of the eosinophilic inflammation activity in bronchial asthma]. / Moznosti sledování aktivity eozinofilního zánetu u asthma bronchiale.

BACKGROUND: Bronchial asthma is a chronic inflammatory disease of the respiratory tract where variety of cells plays a role, particularly mast cells, eosinophils (Eo), and T lymphocytes. At present, there is no clear-cut clinical or laboratory parameter to monitor the activity of this disease. Our study was designed to examine and compare serum eosinophilic cationic protein (S-ECP) levels, plasma ET-1 (P-ET) levels and percentage of eosinophils with CD44 (EoCD44) in paediatric asthmatic patients. METHODS AND RESULTS: In our study, a group of 97 atopic children with persisting mild asthma, had a detailed analysis of their personal history. In addition, S-ECP, P-ET, EoCD44, eosinophil blood count (Eo) and serum levels of IgE(S-IgE) in peripheral blood were determined. Subsequently, children were treated with montelukast (singular), a leukotriene receptor antagonist for a period of three weeks (montelukast tablets in a dose of 5 mg once a day). A second S-ECP, P-ET, EoCD44 were determined in the interval of 3 months from the first collection. In 97 asthmatic children a correlation between P-ET and EoCD44 (p=0.002; r= -0.5) were found. CONCLUSIONS: Our follow-up study surprisingly confirmed a correlation between P-ET and EoCD44. The lower percentage of EoCD44 in peripheral blood in asthmatic children is due to Eo inflammation activity and attests the massive Eo invasion into the airways. The determination of combination - S-ECP, P-ET, EoCD44 - provides an indirect evidence of the multiple features of Eo inflammation.

The use of oscillometry as a measure of airway responsiveness in asthmatic children after histamine and methacholine bronchoprovocation with dosimeter-MedicAid and DeVilbiss nebulizers.

To explore the use of oscillometry as a measure of airway responsiveness, 69 asthmatic children underwent histamine and methacholine bronchoprovocation using dosimeter-MedicAid (Jaeger Co.; Germany) and DeVilbiss nebulizers (DeVilbiss, Bornemouth; England). The mean increase in R5 resistance in challenge testing measured after methacholine with the dosimeter-MedicAid nebulizer was 77.14% compared with 65.05% using histamine. Using the dosimeter-DeVilbiss nebulizer, the mean increases in R5 resistance following methacholine and histamine testing were 57.50% and 59.36%, respectively. The resistance R5 over R20 significantly correlated with forced expiratory volume in 1 second (FEV1). The MedicAid produced a more aggressive challenge than the DeVilbliss nebulizer. Oscillometry can be used to monitor the level of airway hyperresponsiveness following bronchoprovocation tests.