Associations between the presence of specific antibodies to the West Nile Virus infection and candidate genes in Romanian horses from the Danube delta.

The West Nile virus (WNV) is a mosquito-borne flavivirus causing meningoencephalitis in humans and animals. Due to their particular susceptibility to WNV infection, horses serve as a sentinel species. In a population of Romanian semi-feral horses living in the Danube delta region, we have analyzed the distribution of candidate polymorphic genetic markers between anti WNV-IgG seropositive and seronegative horses. Thirty-six SNPs located in 28 immunity-related genes and 26 microsatellites located in the MHC and LY49 complex genomic regions were genotyped in 57 seropositive and 32 seronegative horses. The most significant association (pcorr < 0.0002) was found for genotypes composed of markers of the SLC11A1 and TLR4 genes. Markers of five other candidate genes (ADAM17, CXCR3, IL12A, MAVS, TNFA), along with 5 MHC class I and LY49-linked microsatellites were also associated with the WNV antibody status in this model horse population. The OAS1 gene, previously associated with WNV-induced clinical disease, was not associated with the presence of anti-WNV antibodies.

Genetic susceptibility to West Nile virus infection in Camargue horses.

West Nile virus (WNV) is a mosquito-borne zoonotic neurotropic virus capable to cause lethal meningoencephalitis (WNE) in infected hosts such as birds, horses, and humans. Due to their sensitivity, horses serve as sentinel species in areas at risk. We studied a population of Camargue horses living in Southern France in two zones with endemic WNV circulation where WNV outbreaks were recorded in 2000 and 2003-4. Two sets of microsatellite markers located in MHC and Ly49 genomic regions were genotyped as well as multiple SNPs in ten immunity-related candidate gene regions. Associations between genetic polymorphisms and resistance/susceptibility to WNE were tested. While single marker associations were weak, compound two-gene genotypes of SNPs located within the MAVS, NCR2 and IL-10 genes and microsatellites HMS082 and CZM013 were associated with susceptibility to WNE. Combinations of microsatellite markers CZM009, ABGe17402 and ABGe9019 were associated with simple seroconversion without clinical signs of WNE (resistance). In addition, a distribution of polymorphic markers between WNV-IgG seropositive horses and a control group of WNV-IgG seronegative horses was tested. One SNP in the OAS1 gene (NC_009151.3:g.21961328A>G) was significantly associated with the seropositive phenotype (pcorr = 0.023; OR = 40.5 CI (4.28; 383.26); RR = 8.18 CI (1.27; 52.89) in the Camargue breed. In compound genotypes, SNP markers for SLC11A1, MAVS, OAS1, TLR4, ADAM17 and NCR2 genes and ten microsatellites showed non-random distribution between seropositive and seronegative groups of horses. Further analysis of associated markers could contribute to our understanding of anti-WNV defense mechanisms in horses.

Toxic epidermal necrolysis data from the CELESTE multinational registry. Part I: Epidemiology and general microbiological characteristics.

INTRODUCTION: Toxic epidermal necrolysis (TEN) is a rare, life-threatening autoimmune disease predominantly manifested in the skin and mucous membranes. Today, infectious complications have the dominant share in mortality of TEN patients. Due to the nature of the therapy and administration of immunosuppressive medications, a wide range of potentially pathogenic microorganisms, which cause infectious complications in different compartments in these patients, is not surprising. MATERIAL AND METHODOLOGY: This is a multicentric study, which included all patients with TEN hospitalized between 2000-2015 in specialized centres in the Czech Republic and Slovakia. The total catchment area was over 12.5 million inhabitants. The actual implementation of the project was carried out using data obtained from the registry CELESTE (Central European LyEll Syndrome: Therapeutic Evaluation), when specific parameters relating to epidemiological indicators and infectious complications in patients with TEN were evaluated in the form of a retrospective analysis. RESULTS: In total, 39 patients with TEN were included in the study (12 patients died, mortality was 31%), who were hospitalized in the monitored period. The median age of patients in the group was 63 years (the range was 4-83 years, the mean was 51 years), the median of the exfoliated area was 70% TBSA (total body surface area) (range 30-100%, mean 67%). SCORTEN was calculated for 38 patients on the day of admission. Its median in all patients was 3 (range 1-6; mean 3). Any kind of infectious complication in the study group was recorded in 33 patients in total (85%). In total, 30 patients (77%) were infected with gram-positive cocci, 27 patients (69%) with gram-negative rods, and yeast cells or fibrous sponge were cultivated in 12 patients (31%). A total of 32 patients (82%) were found to have infectious complications in the exfoliated area, 15 patients (39%) had lower respiratory tract infections, 18 patients (46%) urinary tract infections and 15 patients (39%) an infection in the bloodstream. The most common potentially pathogenic microorganism isolated in our study group was coagulase neg. Staphylococcus, which caused infectious complications in 24 patients. Enterococcus faecalis/faecium (19 patients), Pseudomonas aeruginosa (17 patients), Staphylococcus aureus (11 patients) and Escherichia coli (11 patients) were other most frequently isolated micro-organisms. CONCLUSION: The published data were obtained from the unique registry of TEN patients in Central Europe. In the first part, we have succeeded in defining the basic epidemiological indicators in the group of patients anonymously included in this registry. The study clearly confirms that infectious complications currently play an essential role in TEN patients, often limiting the chances of survival. The study also shows a high prevalence of these complications in the period after 15days from the start of hospitalization, when most patients already have completely regenerated skin cover.

Toxic epidermal necrolysis data from the CELESTE multinational registry. Part II: Specific systemic and local risk factors for the development of infectious complications.

The aim of the study was to identify the most important systemic and local risk factors for the development of infectious complications in patients with toxic epidermal necrolysis (TEN). MATERIAL AND METHODOLOGY: This is a multicentric study that included all patients with TEN who were hospitalized between 2000-2015 in specialized centres in the Czech Republic and Slovakia. The total catchment area included a population of over 12.5 million inhabitants. The actual implementation of the project was carried out using data obtained from the CELESTE (Central European LyEll Syndrome: Therapeutic Evaluation) registry, wherein specific parameters related to epidemiological indicators and infectious complications in patients with TEN were evaluated as a retrospective analysis. RESULTS: A total of 38 patients (97%) of the group were treated with corticosteroids. The comparison of patients with different doses of corticosteroids did not exhibit a statistically significant effect of corticosteroid administration on the development of infectious complications (p=0.421). There was no effect of the extent of the exfoliated area on the development of infectious complications in this area. The average extent of the exfoliated area was 66% TBSA (total body surface area) in patients with reported infectious complications and 71% TBSA (p=0.675) in patients without infectious complications. In the case of the development of an infectious complication in the bloodstream (BSI), the increasing effect of the SCORTEN (SCORe of Toxic Epidermal Necrosis) value was monitored during hospitalization. Within 5days from the beginning of the hospitalization, the average SCORTEN value was 2.7 in 6 patients with BSI and 3.0 in 32 patients without BSI (p=0.588). In the period after the 15th day of hospitalization, 7 patients with BSI had an average SCORTEN value of 3.4, and 16 patients without BSI had an average SCORTEN value of 2.5 (p=0.079). In the case of low respiratory tract infection (LRTI), the effects of the necessity for artificial pulmonary ventilation and the presence of tracheostomy were monitored. The statistically significant effect of mechanical ventilation on the development of LRTI occurred only during the period of 11-15days from the beginning of the hospitalization (p=0.016). The effect of the tracheostomy on the development of LRTI was proven to be more significant. CONCLUSION: We did not find any statistically significant correlation between the nature of immunosuppressive therapy and the risk of developing infectious complications. We failed to identify statistically significant risk factors for the development of BSI. Mechanical ventilation and tracheostomy increase the likelihood of developing LRTIs in patients with TEN.

MICROMYCETES INFECTION IN PATIENTS WITH THERMAL TRAUMA.

GOAL: Determination of basic epidemiological parameters of burn patients with micromycetes infection. Identification of the most important micromycetes in burn patients. MATERIAL AND METHODS: Monocentre retrospective study enrolling all adult burn patients who were hospitalized between 2007 and 2015 and in whom micromycetes were isolated during hospitalization. ABSI index (Abbreviated Burn Severity Index) was used to evaluate severity of thermal trauma. Results were statistically analysed. RESULTS: There were 61 patients with thermal trauma identified in total during the period of observation, and there were yeast or fibrous fungi isolated. There were 37 males and 24 females (M:F ratio - 1.5:1) in this group. The average age of patients was 57.3 years (29 patients were aged up to 60 years, 32 patients were over the age of 60 years, inclusive). 6 patients died (lethality was 9.8%). The average extent of the burn area was 21.6% TBSA (median 14.0%). There were 90 strains of micromycetes cultured in total in these patients (79 yeasts, 11 fibrous fungi). Micromycetes were isolated from burn area in 30 patients, from the lower airways in 19 patients, from the urogenital area in 15 patients and from blood culture in 7 patients. Non-albicans Candida species were predominant among yeasts (60 strains); Candida albicans was isolated 16 times in total. Aspergillus fumigatus (4 isolations) and Fusarium species (2 isolations) were predominant species among fibrous fungi. CONCLUSION: We successfully identified the basic epidemiological parameters in burn patients with micromycetes infection, similarly to the most important yeasts and fibrous fungi causing infection in these patients.

Molecular cytogenetic analyses of hTERC (3q26) and MYC (8q24) genes amplifications in correlation with oncogenic human papillomavirus infection in Czech patients with cervical intraepithelial neoplasia and cervical carcinomas.

UNLABELLED: It is known that cervical cancer develop from precancerous intraepithelial neoplasia (CIN) which is characterized by series of genetic abnormalities. The progression of CIN to cervical carcinoma has been associated especially with the genomic integration of oncogenic human papilloma virus (HPV) and gain of the human telomerase RNA gene hTERC (3q26) and MYC (8q24). In this study, cytology specimens of cervical intraepithelial neoplasia and cervical carcinoma from 74 Czech women were analyzed using the triple-color Cervical FISH Probe Kit designed for identification of HPV infected cells and copy number aberration of the hTERC and MYC genes. HPV-positivity exhibited 70% of patients with premalignant lesions (CIN I - CIN III, carcinoma in situ), chromosomal changes were found in 53.3% of cases - MYC amplification had 33.3% of women with CIN I - CIN III and 50% with carcinoma in situ. Amplification of hTERC was detected in 16.7% of patient with CIN I, in 50% with CIN II, in 58.3% with CIN III and in 66.7% with carcinoma in situ. Based on HPV-positivity and the occurrence of chromosomal aberrations, patients were divided into high-, intermediate- and low-risk group. Among women with cervical carcinomas, HPV infection was detected in 90.1% of specimens and chromosomal aberrations were found in 87.5% of samples. Amplification of MYC gene was detected in 25% and hTERC gene in 62.5% of patients. According to the histopathological grade of tumors, MYC gene amplification occurred more frequently in specimens of spinocellular carcinoma than adenocarcinoma (p=0.029). We found no association between the frequency of cytogenetic lesions and the incidence of lymphangiogenesis or lymph node metastases in cervical carcinoma patients. Simultaneous hTERC and MYC genes amplification was significantly more frequent in samples of cervical carcinomas than in premalignant lesions (p=0.008).In a cohort of 26 patients with cervical carcinoma we used oligo-based GGH+SNP microarray technique for the high resolution mapping of copy number changes of hTERC and MYC genes. We found that recurrent gain of genetic material in chromosome 3q26 area carrying hTERC gene of size 43.6 Mb between 3q25.1-3qter and duplication of 3q were the most common genomic identifications of amplified gene. In MYC locus array-CGH profiling identified duplication of 8q and trisomy 8 as frequent genomic changes.Our work confirmed that in cervical carcinoma gains of hTERC and MYC genes are specific genomic changes associated with developing of malignant phenotype. We also showed that in premalignant stages HPV-FISH assay can be used as an effective diagnostic procedure to identify patients carrying highly risking HPV infection and chromosomal aberrations associated with this malignancy. KEYWORDS: cervical cancer, cervical dysplasia, HPV infection, hTERC amplification, MYC amplification, FISH, array-CGH.

ATM and TGFB1 genes polymorphisms in prediction of late complications of chemoradiotherapy in patients with locally advanced cervical cancer.

The purpose of our study was to evaluate a possible correlation between genetic polymorphisms in ATM and TGFB1 genes and late toxicity of chemoradiotherapy for locally advanced cervical cancer. Fifty five patients with FIGO stage IIB and higher without a disease recurrence with a mean follow up of 6 years were included. Late toxicity was assessed by EORTC/RTOG late toxicity criteria. Univariate and multivariate logistic regression model was used for statistical analysis. Degree of association between polymorphisms and late toxicity of chemotherapy was assessed on the basis of phi-coefficient (φ) as well. We did not find any association between 5557G>A polymorphism in the ATM gene or single TGFB1 polymorphisms and late toxicity. TGFB1 compound homozygosity (-1552delAGG, -509C>T, L10P) was a significant predictive factor of grade III-IV and any grade of complications in both univariate and multivariate logistic regression analyses and statistical significance of association between polymorphisms and late toxicity of chemoradiotherapy was confirmed also by the evaluation of phi-coefficient (φ). We conclude that haplotypes instead of single nucleotide polymorphic sites in the genes may better characterize the individual radiosensitivity.

Effect of timing of tracheostomy on changes in bacterial colonisation of the lower respiratory tract in burned children.

OBJECTIVE: The study aims to evaluate the impact of early and late tracheostomy on microbiological changes in the airways in severely burned children. MATERIALS AND METHODS: Early tracheostomy is sometimes performed within 3 days after the start of mechanical ventilation regular microbiological surveillance of the respiratory tract was done in all patients. From each sputum, tracheobronchial aspirate and bronchoalveolar lavage (BAL), a microscopic slide was made and the material was seeded in a culture medium. The standard culture media used for the growth of respiratory pathogens are blood agar, McConkey agar, VL agar and chocolate agar. The obtained values were statistically analysed. RESULTS: In the observed period, a total of 68 children underwent mechanical ventilation in our department. A total of 31 (45.59%) children had undergone surgical tracheostomy (18 patients with early tracheostomy and 13 patients with late tracheostomy). The most common bacterium isolated from the lower respiratory tract in patients with early and late tracheostomy was Acinetobacter baumannii (31.53% resp. 44.30% of all bacterial strains). In patients with early tracheostomy, the ratio of G+/G- during the 6-7th day of mechanical ventilation was 1.29:1 and during the 8-10th day, 1:1.43. In patients with late tracheostomy the G+/G- ratio was 1:2.25 and during the 8-10th day, 1:2.25. There was not any statistically significant deviation in the G+/G- ratio in patients with early and late tracheostomy in any of the monitored periods. CONCLUSION: The main reasons for performing early tracheostomy are: extent, localisation and depth of the burn. Difficult weaning in an uncooperative patient, failure of extubation with subsequent reintubation and other complications may be an indication for late tracheostomy. The study confirms that the use of appropriately indicated early tracheostomy provides a microbiological benefit for burned children.

[Epidemiology of critical burns in southern Moravia]. / Epidemiologie kritických popálenin na jizní Morave

UNLABELLED: The aim of the study was to define the principal and additional characteristics of critical burns and to evaluate the effect of an increasing Abbreviated Burn Severity Index (ABSI) on mortality in burn patients admitted to the Department of Burns and Reconstructive Surgery, University Hospital in Brno. MATERIAL AND METHODS: This retrospective monocentric study included all patients older than 18 years with critical burns involving more than 40% of the total body surface area (TBSA) who were admitted to the Department of Burns and Reconstructive Surgery, University Hospital in Brno, in the period from January 1, 2001 to December 31, 2010. The epidemiological data evaluated included: age, gender, size of burn injury, mechanism of injury, required surgical intervention, mortality, length of hospital stay and the presence of inhalation injury. The results were statistically analysed. RESULTS: A total of 2 479 adult patients were admitted to the Department in the period under study, and 112 (4.5%) of them had critical burns. In this patient group, the average age was 48.7 years, the average burned surface area was 62.8% TBSA, and the average length of hospital stay was 37.2 days. The male-to-female ratio was 2.39:1. Inhalation injury was diagnosed in 92 patients (82.1%). Of the 112 patients with burn injury, 59 died (52.7%). The lowest mortality rate was in the age category of 31 to 40 years (21.1%) and the highest rate in the patients aged over 71 years. No patient with the ABSI . 8 died while, amongst the patients with the ABSI .13, the mortality rate was 87.8%. DISCUSSION: No study dealing comprehensively with this topic in a patient group of this size has so far been published in the Czech Republic. Our results showed that the patients with critical burns accounted for about 5% of all patients with thermal trauma. The ABSI has proved to be a valuable and reliable predictor of survival in patients with critical burns. The extent of burned area and age were risk factors affecting mortality. On the other hand, the effect of inhalation injury on mortality, as a single risk factor, is disputable because this develops with an increasing burned surface area. CONCLUSIONS: The most important epidemiological data on patients with critical burns were evaluated. The study shows that although the number of patients annually admitted to hospital with severe burns is still high, it has nevertheless decreased recently.

[Genetic background of cisplatin induced ototoxicity]. / Genetické pozadí ototoxicity cisplatiny.

BACKGROUND: Cisplatin induced ototoxicity is a serious adverse effect of cisplatin therapy. Cisplatin induced ototoxicity shows significant interindividual variability. This variability is probably based on genetic background. Recent papers describe association of cisplatin ototoxicity with allelic variants of glutathion-S-transferase coding genes. PATIENTS AND METHODS: We have analyzed 55 patients treated with cisplatin therapy without any previous hearing impairment. Audiometric examination was performed before the start of cisplatin therapy and then before and after each cycle of cisplatin. DNA isolated from peripheral blood samples was used to analyze genetic polymorphisms of selected genes coding for glutathion-S-transferases. RESULTS: We have demonstrated association of early onset of cisplatin induced hearing impairment with absence of null allele of GSTT1 (p = 0.009). Both GSTM1 gene deletion and single nucleotide polymorphism in GSTP1 gene (rs1695) did not show any association with cisplatin induced ototoxicity. CONCLUSION: Early onset of cisplatin induced hearing impairment is more probable in persons with two functional alleles of GSTT1 gene.

[Femoral lengthening. A critical review of ten-year results]. / Kritické zpracování prodluzování femuru za posledních deset let.

PURPOSE OF THE STUDY: To evaluate the results of femoral lengthening in the patients treated from 2000 to 2009 in whom complete radiographic data were available and the lengthening procedure involved mainly the use of a Mephisto fixator. MATERIAL AND METHODS: A total of 28 femoral lengthening procedures were carried out in 26 patients,16 girls and 10 boys. The external fixator Mephisto was used in 24 cases; fixators Prospon and Orthofix in one and three cases, respectively. Fifteen patients were treated for congenital short femur, the rest had secondary femoral shortening due to following pathologies: distal femoral epiphysiolysis in five children, proximal femoral osteomyelitis in one child, avascular necrosis of the femoral head in one, diaphyseal femur fracture in one, enchondromatosis of the distal femur with growth plate destruction in one, and contralateral femur overgrowth following a fracture in one child. The average age at the beginning of treatment was 11 (range, 4-16) years. Complications were classified as mild, serious and critical. The results were statistically analysed using several statistical tests. RESULTS: The average parameter values for the group included: total femoral lengthening, 40.2.mm (SD±11.1); osteotomy index (OI), 41 % (SD±9.8); lengthening percentage (LP), 10.9 % (SD±3.8); lengthening index (LI), 14.5 (SD±3.5) days/cm; hea- ling index (HI), 52.6 (SD±20.1) days/cm; and consolidation index (CI), 93.3 (SD±40.0) days/cm. Mild complications were recorded in 11 (39.2 %), and serious and critical in eight patients (28.6 %). Fourteen patients (53.8 %) were free of any complications. Two complications were concurrently found in five patients (17.9 %). There was a statistically significant difference in the LP values related to the number of complications (p=0.019). No significant relationship was recorded on comparison of the HI value with the patient's age at the time of surgery (p=0.836) and patient's gender (p=0.546) (Mann- Whitney test). The relationship of the OI value to the HI value was non-significant (p=0.492), as was the relationship between the osteotomy technique (oscillating or Gigli saw osteotomy) and the occurrence of complications (p=1.000) (Fisher's exact test). Correlation between the LI and HI values was significant (p<0.001). DISCUSSION: The results of healing after lengthening, as assessed by the healing and the consolidation index, were in agreement with other authors' data. The lower number of complications, particularly fractures of bone regenerate, can be explained by the facts that, in our study, the lengthening percentage was lower and that the post-operative care was strictly observed, including dynamic axial loading which stimulates bone consolidation at the lengthened section, with adherence to the proof of three developed cortices. CONCLUSIONS: Our results did not confirm the assumption that slower lengthening will have a favourable effect on the healing index. Key words: femoral lengthening, external fixator, complications.

[Blood lipid changes at long-term antiretroviral treatment]. / Zmeny krevních lipidu pri dlouhodobé antiretrovirové lécbe.

UNLABELLED: The aim of study was to find the development trend of blood lipid concentration in a group of HIV positive patients treated by combination antiretroviral therapy (cART). We followed changes during the therapy and evaluated their aterogennic nature. METHODS: The group included 118 patients stepwise allocated to the AIDS Centre of the Faculty Hospital Brno, with the monitoring period being up to 1 month as the minimum and up to 17 years as the maximum. The patients were divided into cART treated patients and not treated patients. The following parameters were analysed: total cholesterol, triglycerides, HDL-cholesterol, apolipoprotein B, the total cholesterol/HDL-cholesterol index and non-HDL-cholesterol. RESULTS: Our group experienced a statistically significant increase of total cholesterol concentration already in the first months after cART initiation and this value continuously increased in the following years. The recommended target value for total cholesterol (5 mmol/l) was exceeded in the group of patients after 3-4 years of cART initiation. The triglyceride concentration showed a sudden increase already a few months after cART initiation, when the recommended optimum value of triglycerides (1.7 mmol/I) was exceeded. These changes had a further no statistic significance. The average triglyceride value was all around (slightly above) 1.7 mmol/l. Our group experienced a statistically significant increase of HDL-cholesterol concentration in the first two years after cART initiation. A statistically significant change of HDL-cholesterol concentration was not found in the following years. The average HDL-cholesterol value was above optimal value HDL-Ch > 1.0 mmol/l for men (except initial category). A statistically significant change of apolipoprotein B concentration was found after 3-4 years of cART treatment. However, the average apolipoprotein B value did not exceed the target value in any of the followed categories. No statistically significant changes of the total cholesterol/HDL-cholesterol index were found. The resulting value was under 5 in all the followed categories. Statistically significant changes of non-HDL-cholesterol were found in patients with cART already a few months after treatment initiation and its concentration continually increased. However, the recommended target value of non-HDL-cholesterol (3.8 mmol/l) was exceeded only in the category of patients treated 4 - 5 years. The development trend of CD4+ lymphocyte count and HIV-1 RNA copies means high active of cART from standpoind of immunoregeneration (CD4+ lymphocyte count) and viral suppression (HIV-1 RNA copies) even in the group of treated patient with the longest monitoring period. CONCLUSION: Monitoring of our group of HIV-positive patients treated by combination antiretroviral therapy revealed a statistically significant increase of blood lipid concentrations (inclusive of HDL-cholesterol) during the treatment. However, these changes do not have an unequivocally aterogennic nature even in the group of treated patient with the longest monitoring period.