The effect of warfarin administration on platelet aggregation.

BACKGROUND: The aim of this study was to assess the effect of anticoagulation treatment on platelet aggregation. METHODS: The study group consisted of 24 patients on long-term warfarin therapy without any antiaggregation therapy. Platelet aggregation was measured using VerifyNow with arachidonic acid (AA) as an inducer in 23 patients and with light transmission aggregometry (LTA) in 19 patients using four different agonists. All patients had their international normalized ratio (INR) checked regularly. RESULTS: The mean INR value was 2.07 (SD 0.6). The average aggregation measured by VerifyNow was found to be 637.5 (SD 36.6) aspirin reaction units. The values of average aggregability in LTA were 73.3 % (SD 4.5 %), 73.2 % (SD 6 %) and 72.1 % (SD 4.8 %) in case of aggregation induced by AA, ADP, and collagen, respectively. Epinephrine­induced aggregability was 65.3 % (SD 14.7 %). Regression analysis between INR and values of collagen- or epinephrine­induced aggregability (r = 0.654 and 0.575) was found statistically significant (p = 0.004 and 0.016); every increase in INR by 0,1 brings about an increase in collagen- and epinephrine­induced aggregation values by 1.5 and 4, respectively. CONCLUSION: Administration of warfarin does not produce a significant reduction in platelet aggregation. On the contrary, prolonged INR evokes a mild increase in aggregation induced by collagen or epinephrine (Tab. 2, Fig. 3, Ref. 32). Text in PDF www.elis.sk Keywords: platelet aggregation, anticoagulation, warfarin, platelet function tests, chronic ischemic heart disease.

The use of estradiol metered-dose transdermal spray in clinical practice.

OBJECTIVE: The aim of this study was to evaluate the use of novel estradiol metered-dose transdermal spray (EMDTS) in the treatment of acute climacteric syndrome. METHODS: A multicenter open-label trial was conducted with a 24-week intervention. EMDTS 1.53 mg was given to symptomatic menopausal women. The Menopause Rating Scale (MRS) was used to assess the climacteric syndrome severity. The Friedman non-parametric test and a post-hoc test with Bonferroni correction were used for statistical evaluation. RESULTS: A total of 132 women were enrolled in 20 centers, of whom 123 (93.2%) completed the study. The average age of patients was 53.8 years (37-65 years). The study was discontinued by 6.8% of women. The patients were checked at the beginning of the study, and after 12 and 24 weeks. There was a statistically significant drop (p < 0.001) in MRS values both after 12 and 24 weeks of therapy. The average MRS values improved by 66.2% between the first and the third visits. The most significant improvement was manifested in patients with initial moderate climacteric syndrome (70.9%). CONCLUSION: This study confirms that application of EMDTS offers a novel treatment option for climacteric symptoms.

Comparison of the epidemiological patterns of Lyme borreliosis and tick-borne encephalitis in the Czech Republic in 2007-2016.

BACKGROUND: Lyme borreliosis (LB) and tick-borne encephalitis (TBE) are two vector-borne diseases transmitted by ticks. Both diseases are endemic and have been reported in all regions of the Czech Republic including urban agglomerations, but in varying proportions. Because of the natural conditions in the border areas, the risk of infection is also high for travelers from the neighboring countries such as Austria, Germany, Poland, and Slovakia. MATERIALS AND METHODS: To gain more information on the epidemiology of LB and TBE in the last decade, we analyzed national surveillance data from 2007 to 2016. RESULTS: Incidence of LB in the Czech Republic was 37.3/100,000 population and year (27.6 - 46.1/100,000). Incidence of TBE incidence was 5.7/100,000 (3.4-8.2/100,000) and declined although the trend was not significant (p = 0.155). Difference between the incidences of LB and TBE was increasing in time. Overall male-to-female ratio was 0.84 and 1.51 for LB and TBE, respectively. The age-specific incidences of both infections have a typical two-peak shape, with the first peak in the age group 5-9 years for LB and 15-19 years for TBE. The second peak for both LB and TBE is in the age group 55-64 years. TBE poses a considerable risk to children < 15 years. Among 39,074 cases of LB, the most common clinical manifestations were erythema migrants 62.1% and Lyme neuroborreliosis 25.1%. All 5969 TBE cases manifested itself by affecting nervous system, namely meningo-encephalitis 47.9%, meningoencephalomyelitis 21.8% and meningitis 19.8%. CONCLUSIONS: The data evidence the high chance risk of infection with LB and TBE in the Czech Republic. The incidence of both infections shows a bimodal distribution during the year. LB cases are five to six times as frequent as TBE cases. Over the last years, the incidence of LB has remained roughly stable while TBE has shown a downward trend. The present study is unique in allowing the comparison of the incidence rates of LB and TBE over time and space.

Melanoma antigens are biomarkers for ipilimumab response.

BACKGROUND: Novel immunotherapy modalities significantly improve survival of patients with metastatic melanoma. However, CTLA-4-blocking monoclonal antibody ipilimumab is effective only in a small proportion of patients. Biomarkers for prediction of treatment response are indispensably needed. OBJECTIVE: To determine the utility of multimarker detection of circulating melanoma cells as prognostic and pharmacodynamic biomarker in patients with metastatic melanoma treated with ipilimumab. METHODS: Patients (n = 62) with metastatic melanoma in unresectable stage III or metastatic stage IV treated with ipilimumab were recruited prospectively. The values of four melanoma markers on circulating cells Melan-A, gp100, MAGE-3 and melanoma inhibitory antigen prior to the treatment and within the therapy were compared to the data collected at baseline - after the melanoma surgery. RESULTS: The immunotherapy pretreatment marker level was found to be prognostic of overall survival; lower levels were linked to longer survival time. Moreover, longitudinal follow-up of melanoma markers in patients treated with ipilimumab correlates with therapy response. A decline of marker levels by >30% at week 6 (in 83% of the responding subjects) to week 9 (in all responders) of ipilimumab administration was associated with response to therapy. Elevation of the tumour markers during the treatment precedes clinical progression and gives an early warning of treatment failure. CONCLUSION: Melanoma circulating cells hold potential as predictive and pharmacodynamic biomarker of immunotherapy.

The occurrence of Ixodes ricinus ticks and important tick-borne pathogens in areas with high tick-borne encephalitis prevalence in different altitudinal levels of the Czech Republic Part II. Ixodes ricinus ticks and genospecies of Borrelia burgdorferi sensu lato complex.

STUDY OBJECTIVE: Three years long research study (2011-2013) on population density of Ixodes ricinus and the infection rate of the pathogens that they transmit was conducted in four topographically distant areas in the Czech Republic. In the previous decade (2001-2010) thirteen loci with increased incidence of tick borne encephalitis cases were defined, suggesting the permanent interaction of human population with ticks and indicating the landmarks for study of the presence of other tick borne pathogens. The work program included the identification of existing spectrum of spirochetes from Borrelia burgdorferi sensu lato complex and the conditions of their occurrence and distribution. MATERIAL AND METHODS: In the areas of the Ústí nad Labem Region, Olomouc Region, South Bohemian Region, and Highlands Region, 600 m2 plots were selected in the local optimal I. ricinus habitats where tick flagging was performed every year in the spring-summer and autumn seasons of the tick questing activity. Collected adult ticks (1369 males and 1404 females) were individually screened for B. burgdorferi s. l. spirochets. RESULTS: Spirochetes from B. burgdorferi s.l. complex were detected in all 13 studies sites in all altitudes from 280 to 1030 meters a. s. l. The total rate of infection was determined as 11.4% (males 10.4%, females 12.4%) with range limits from 1.4% (Ústí nad Labem in 2011) to 19.7% (South Bohemian Region, 2012).Genospecies were detected in various proportions and in different combinations: Borrelia afzelii, B. garinii, B. burgdorferi s. s., B. bavariensis, B. bissettii, B. valaisiana, B. spielmanii and B. lusitaniae. The three-year observation justifies the assumption that the regional differences in infectivity of I. ricinus are based on the character of the local biocenosis of the respective region. The dynamics of its seasonal changes, conditioned by climatic factors, determines the annual differences. CONCLUSION: Three of the medically most important Borrelia species formed a core group among all detected genospecies. B. afzelii was a dominated one (115 detections), followed by B. garinii (100) and by B. burgdorferi s.s. (19). Other genospecies were detected sporadically. However, the detection of B. bissettii should be emphasized due to the recently proven pathogenic effects of this genospecies and yet little-known sporadic expansion in the Czech Republic. The medical importance and distribution of other sporadically occurred genospecies is also discussed.Key words: Ixodes ricinus - Borrelia afzelii - B. garinii - B. burgdorferi s. s. - B. bavariensis - B. valaisiana - B. spielmanii - B. lusitaniae - B. bissettii - distribution - altitude - season - medical importance.

The occurrence of Ixodes ricinus ticks and important tick-borne pathogens in areas with high tick-borne encephalitis prevalence in different altitudinal levels of the Czech Republic Part I. Ixodes ricinus ticks and tick-borne encephalitis virus.

STUDY OBJECTIVE: The aim of the three-year study (2011-2013) was to monitor population density of Ixodes ricinus ticks and its infection rate with the tick-borne encephalitis virus in areas with a high incidence of tick-borne encephalitis as reported in the previous decade 2001-2010. Such a comprehensive and long-term study based on existing epidemiolo-gical findings has not previously been conducted in Europe. MATERIAL AND METHODS: In the areas of the Ústí nad Labem Region, Olomouc Region, South Bohemian Region, and Highlands Region, 600 m2 plots were selected in the local optimal I. ricinus habitats where tick flagging was performed every year in the spring-summer and autumn seasons of the questing activity. In total, 18,721 I. ricinus ticks (1448 females, 1425 males, and 15,848 nymphs) were collected and investigated. RESULTS AND CONCLUSION: The results have shown that the differences in the infection rate of I. ricinus observed between regions are driven by variation in the density of the local I. ricinus populations which is influenced by the characteris-tics of the whole local biocenosis. The overall prevalence estimate of TBE virus in Ixodes ricinus ticks at the altitudes below 600 m a.s.l. was 0.096 % (95% CI 0.055-0.156) for nymphs, and 0.477 % (95% CI 0.272-0.773) for adults. The dynamics of the seasonal variation in I. ricinus populations, depending primarily on the climatic factors, are behind the interyear differences in the infection rate of ticks and, consequently, in the epidemiological situation of tick-borne encephalitis. The nymph to adult ratio was 5.5 on average but showed great interregional variability (from 10.3 in the Ústí nad Labem Region to 1.8 in the Highlands Region). It might be used in the future as one of the indicators of the composition of the local I. ricinus population and of the level of the circulation of tick-borne pathogens in zoonotic sphere and also for use in the health risk assessment in a given area. Despite the permanent expansion of ticks and tick-borne pathogens in higher altitudes the high risk limit for human infection with tick-borne encephalitis is 600 m a.s.l. in the Czech Republic.

[Immune System in Patients with Head and Neck Carcinoma]. / Imunitní systém u pacientu s nádory hlavy a krku.

The insight into the biological nature of head and neck squamous cell carcinoma has evolved significantly in the last few years. Tobacco use and alcohol consumption are proven risk factors of head and neck squamous cell carcinoma. Cancer patients possessing such a tumor are generally elderly, mostly in fifth or sixth decade of life. In addition, significant association of head and neck squamous cell carcinoma with infection by human papillomavirus (HPV) was proven. HPV is now considered to be one of the most important risk factors, particularly for oropharyngeal carcinoma. HPV  positive tumors of oropharynx are associated with significantly better prognosis. Experimental and clinical data indicate that HPV positive and HPV negative tumors can be considered as two different entities and it has not been clarified which factors are crucial for better prognosis of HPV  positive tumors. The character of immune reaction, which contributes to distinct prognosis, may be one of the important factors. This review summarizes current knowledge concerning various aspects of antitumor immune responses in HPV  positive and HPV  negative tumors. Recent studies have shown that a broad repertoire of tumor infiltrating HPV specific T-cells is detectable in almost all patients with HPV positive tumors. Despite this, there is a development of tumor, which may be facilitated by abnormalities in antigen processing, T-cell dysfunction or prevalence of immunosuppressive cells. Nonetheless, the immunologic profile of HPV positive vs. HPV negative head and neck squamous cell carcinoma is associated with better outcome, and HPV  specific immune response is suggested to play an essential role in the better response to conventional therapy of HPV  positive patients. We also discuss HPV specific antitumor immunotherapy approaches, which are now tested in clinical trials.

[Vulvar melanoma]. / Melanom vulvy.

OBJECTIVE: Malignant melanoma of the vulva is the second commonest vulval malignancy. This article will focus on three cases of vulvar melanoma which have been solved surgically with wide excision, sentinel lymph node biopsy and radical lymphadenectomy. DESIGN: Case report. SETTING: Department of Dermatovenerology 3rd Faculty of Medicine, Charles University, Faculty Hospital Královské Vinohrady, Prague. METHODS: Biopsy of the lesion creates a reliable diagnostic procedure. More frequently digital dermoscopy is used for the precise primary diagnostics and follow-up of pigment vulvar lesions. RESULTS: Vulvar melanoma has been variously estimated to account for between 3.6 and 10% of malignant vulvar neoplasms. An epidemiologic study revealed ratio of vulvar to skin melanoma 1:71. CONCLUSION: New diagnostic methods such as digital dermoscopy or sentinel node biopsy bringing ever greater progress in precise diagnosis of patients with vulvar melanoma.

[K-ras mutational status and tumour-infiltrating lymphocytes in human colon cancer: state of the art and future perspectives]. / K-ras mutace a nádory infiltrující lymfocyty u karcinomu kolon, soucasnost a výhledy.

INTRODUCTION: Nowadays, the prognosis of newly diagnosed colorectal cancer patients relies mostly on the tumour-node-metastasis (TNM) classification which is also a determining criterion for the indication of adjuvant oncological treatment. Currently, new prognostic and predictive biomarkers are sought after in order to more precisely define prognosis and better predict the benefits of adjuvant treatment in colorectal cancer. Besides several molecular biomarkers, such as mutations in the proto-oncogene K-ras, analyses of tumour-infiltrating lymphocytes have shown promising prognostic value. The aim of the study is to examine the correlations between K-ras mutational status and tumour-infiltrating immune cells in colon cancer patients with respect to colon cancer recurrence. MATERIAL AND METHODS: Formalin-fixed paraffin-embedded specimens were obtained from 44 patients with surgically resected colon cancer (R0 resection) treated between 2004 and 2009. K-ras mutational status was detected using PCR amplification of exon 1 followed by direct sequencing and K-ras StripAssay. Tumour-infiltrating immune cells were detected by immunofluorescence staining using monoclonal antibodies against CD3, CD8, FoxP3, CD1a and DC-LAMP. RESULTS: All 44 patients in our cohort underwent radical resection of colon cancer. In 16 patients the tumour relapsed (36.4%). K-ras mutations were found in 45.5% (n=20) of the primary carcinomas: 65% in codon 12 and 35% in codon 13. Although codon 13 K-ras mutations were associated with disease relapse, they were present in both disease-free and relapsed patients. However, disease-free and relapsed patients differed markedly in their patterns of tumour-infiltrating immune cells. There was a trend towards decreased density of tumour-infiltrating lymphocytes within the group of relapsed patients. In addition, relapsed patients with codon 13 mutations had markedly lower levels of tumour-infiltrating mature DC-LAMP+ dendritic cells and higher frequency of CD1a+ cells compared to disease-free patients. CONCLUSION: Colon cancer patients with low levels of tumour-infiltrating lymphocytes, a high CD1a+/DC-LAMP+ tumour-infiltrating DC ratio and a K-ras mutation in codon 13 are at a high risk of disease recurrence.

Comparison of yeast (Candida maltosa) and bacterial (Rhodococcus erythropolis) phenol hydroxylase activity and its properties in the phenolic compounds biodegradation.

Aromatic contaminants of the environment, to which belongs phenol and its derivatives, are toxic and in most of the cases hard to degrade. Removal of these pollutants by biological, gentle and effective way, depends on specific environmental conditions in the locality and on the biodegradation potential of the used microbial population. Closer characterization of the biodegradation and enzyme mechanisms is therefore an essential assumption of the successful implementation of microbes. This paper is focused on comparison of the biodegradation activity between the soil yeast Candida maltosa and bacteria Rhodococcus erythropolis towards various aromatics connected with determination of the first enzyme of the phenol biodegradation pathway: phenol hydroxylase (PH). The effect of substrate type, substrate concentration, growth phase of the microorganisms and presence of humic acids in the cultivation medium, on phenol biodegradation and PH activity are discussed.