16S rRNA gene-based identification of cultured bacterial flora from host-seeking Ixodes ricinus, Dermacentor reticulatus and Haemaphysalis concinna ticks, vectors of vertebrate pathogens.

A total of 151 bacterial isolates were recovered from different developmental stages (larvae, nymphs and adults) of field-collected ticks (67 strains from Ixodes ricinus, 38 from Dermacentor reticulatus, 46 from Haemaphysalis concinna). Microorganisms were identified by means of 16S rRNA gene sequencing. Almost 87 % of the strains belonged to G(+) bacteria with predominantly occurring genera Bacillus and Paenibacillus. Other G(+) strains included Arthrobacter, Corynebacterium, Frigoribacterium, Kocuria, Microbacterium, Micrococcus, Plantibacter, Rhodococcus, Rothia, and Staphylococcus. G(-) strains occurred less frequently, comprising genera Advenella, Pseudomonas, Rahnella, Stenotrophomonas, and Xanthomonas. Several strains of medical importance were found, namely Advenella incenata, Corynebacterium aurimucosum, Microbacterium oxydans, M. schleiferi, Staphylococcus spp., and Stenotrophomonas maltophilia. Data on cultivable microbial diversity in Eurasian tick species D. reticulatus and H. concinna are given, along with the extension of present knowledge concerning bacterial flora of I. ricinus.

[The effect of preoperative administration of heparin on the heparin resistance development]. / Vliv predoperacního podání heparinu na vznik heparinové rezistence.

BACKGROUND: Heparin resistance is relatively frequent problem in cardio surgery. We were try to determine real occurrence heparin resistance before operation. AIM OF THE STUDY: Purpose of the project--to find the real frequency of heparin resistance in patients who will undergo a cardio surgical operation. To find out the dependence between the pre-operational application of heparin and the development of heparin resistance. METHODS: We recorded pre-operative administration heparin in patients. If the dose of heparin was 5 mg/kg and more then we insert patients to the group heparin resistant. RESULTS: In our collection was heparin resistance in 203 patients from 624, it was 32.5%. Test agreement relative frequency with 22% was throw out--p < 0.001--heparin resistance in our group statistically different from 22% heparin resistance was higher than hypothesis. Heparin before operation was administrate 181 patients, which make to 29%. For administration of heparin was hypothesis of independence thrown (p < 0.001). CONCLUSION: Results of our works confirmed statistically significant occurrence of heparin resistance in patients that was administration heparin pre-operative. Heparin resistance occurred against presumption 22% in 32.5% in our group. It is statistic significant difference.

Lactobacillus spp. associated with early childhood caries.

A group of 69 lactobacilli was isolated from caries lesions and root canals of early childhood caries (ECC) affected children treated in the Department of Pedodontics (Children's Teaching Hospital, Brno, Czech Republic). Biochemical and physiological properties of all strains were characterized by API 50 CH kit and conventional tube tests. The rep-PCR fingerprinting with the (GTG)(5) primer was used for genotypic grouping of the isolates. The (GTG)(5)-PCR fingerprinting grouped all analyzed strains into a few clusters in nearly full agreement with phenotype identification results and clarified the taxonomic position of 13 biochemically unidentified strains. In total, 20 strains of Lactobacillus fermentum, 17 L. rhamnosus, 14 L. casei/paracasei, 7 L. gasseri, 7 L. salivarius and 4 L. plantarum were identified. Mixtures of two or even three Lactobacillus spp. were isolated from a few root canal content samples. Results obtained by biotyping and (GTG)(5)-PCR were generally comparable except for L. gasseri strains that were not biochemically identified. The (GTG)(5)-PCR fingerprinting was shown to be quicker, easier to perform and more reliable than biotyping. Our results imply this molecular method as a good tool for screening and identification of Lactobacillus spp. inhabiting dental plaque.

Characterization of Aeromonas encheleia strains isolated from aquatic environments in the Czech Republic.

AIMS: Characterization and identification of Aeromonas strains isolated from surface and underground waters using phenotypic and genotyping methods. METHODS AND RESULTS: Biotyping using the ENTEROtest 24 kit and conventional biochemical and physiological tests assigned four strains to Aeromonas encheleia, whereas three isolates were identified as ambiguous Aeromonas bestiarum/Aeromonas caviae and one strain as Aeromonas eucrenophila/Aeromonas encheleia. Further characterization grouped the analysed strains together with Aer. encheleia CCM 4582(T) and assigned the analysed group as members of Aer. encheleia species using ribotyping, whole-cell protein analysis and ERIC-PCR fingerprinting. The results obtained were verified by DNA gyrase A subunit gene sequencing. All analysed isolates showed unique molecular patterns, except for isolates P 1769 and CCM 7407, which revealed the same EcoRI ribotype profile and proved to be identical strains. CONCLUSIONS: Our results imply that Aer. encheleia strains occur in unpolluted surface as well as in underground waters and demonstrate applied methods as suitable for their identification. SIGNIFICANCE AND IMPACT OF THE STUDY: To our best knowledge, this is the first report of the isolation and identification of Aer. encheleia in the Czech Republic.

HLA DRB1, DQB1 and insulin promoter VNTR polymorphisms: interactions and the association with adult-onset diabetes mellitus in Czech patients.

Both the human leucocyte antigen (HLA) DRB1 and the HLA DQB1 gene loci play a role in the development and progression of autoimmune diabetes mellitus (T1DM). Similarly, the insulin promoter variable number tandem repeats (INS-VNTR) polymorphism is also involved in the pathogenesis of diabetes mellitus (DM). We studied the association between each of these polymorphisms and DM diagnosed in patients older than age 35 years. Furthermore, we analysed possible interactions between HLA DRB1/DQB1 and INS-VNTR polymorphisms. Based on C-peptide and GADA levels we were able to distinguish three types of diabetes: T1DM, latent autoimmune diabetes in adults (LADA) and T2DM. INS-VNTR was genotyped indirectly by typing INS-23HphI A/T polymorphism. The genotype and allele frequencies of INS-23HphI did not differ between each of the diabetic groups and group of healthy subjects. We did, however, observe an association between the INS-23HphI alleles, genotypes and C-peptide secretion in all diabetic patients: A allele frequency was 86.2% in the C-peptide-negative group vs. 65.4% in the C-peptide-positive group (P(corr.) < 0.005); AA genotype was found to be 72.4% in the C-peptide-negative group vs. 42.6% in the C-peptide-positive groups (P(corr.) < 0.01). The HLA genotyping revealed a significantly higher frequency of HLA DRB1*03 allele in both T1DM and LADA groups when compared to healthy subjects: T1DM (25.7%) vs. control group (10.15%), odds ratio (OR) = 3.06, P < 0.05; LADA (27.6%) vs. control (10.15%), OR = 3.37, P < 0.01. The simultaneous presence of both HLA DRB1*04 and INS-23HphI AA genotype was detected in 37.5% of the T1DM group compared to only 9.2% of the healthy individuals group (OR = 5.9, P(corr.) < 0.007). We summarize that in the Central Bohemian population of the Czech Republic, the INS-23HphI A allele appears to be associated with a decrease in pancreatic beta cell secretory activity. HLA genotyping points to at least a partial difference in mechanism, which leads to T1DM and LADA development as well as a more diverse genetic predisposition in juvenile- and adult-onset diabetes. The simultaneous effect of HLA and INS-VNTR alleles/genotypes predispose individuals to an increased risk of diabetes development.

Ribotyping and biotyping of Staphylococcus epidermidis isolated from hospital environment.

The Staphylococcus strains acquired from scrapings from hospital environments were identified to the species level based on their biochemical properties. From the monitored sample the Staphylococcus epidermidis strains were selected for more accurate typing and tested on their virulence factor and ribotyped. The biotyping of S. epidermidis did not show any considerable intraspecific variation of these isolates and there were no atypical reactions, with the exception of three strains (out of 33). In contrast, the results of ribotyping showed greater heterogeneity of strains and unequivocally demonstrated the relation between the ribotype and the place of sample drawing. In addition to this fact, the found ribotypes repeat in the same environment in the long-term which suggests the occurrence and persistence of the same strains of conditionally pathogenic bacteria in hospital environment. We showed that ribotyping is a suitable method for precise and reliable detection of some coagulase-negative staphylococci.

KCNJ11 E23K polymorphism and diabetes mellitus with adult onset in Czech patients.

In this work, we studied the association of the E23K polymorphism of the Kir6.2 ATP-sensitive potassium channels in 212 Czech patients with diabetes mellitus who were diagnosed after the age of 35. Patients were classified into T1DM, LADA and T2DM groups based on C-peptide and GADA levels. Carriers of the predisposing Kir6.2 E23K K allele showed no increased risk of either type of diabetes mellitus development. On the other hand, we found a correlation between E23K SNP of the KCNJ11 gene and C-peptide levels, which may be considered a measure of pancreatic beta-cell activity, although this correlation was not statistically significant. In conclusion, we failed to confirm the Kir6.2 E23K as a genetic marker for T1DM, LADA and T2DM in the Central Bohemian population of the Czech Republic.

[Mechanisms influencing implantation of the embryo--the last-years' news]. / Mechanismy ovliviiující implantaci embrya--co je nového v posledních letech.

OBJECTIVE: To summarize recent knowledge concerning mechanisms which influence the implantation of embryo. DESIGN: Literature-based overview. SETTING: Institute for the Care of Mother and Child, Prague. SUBJECT OF THE STUDY: Factors influencing implantation of embryo in the uterus elicit increased interest due to study of unexplained failures of embryotransfer following the successful in vitro fertilization. Our article points to recent information about physiology and pathology of mechanisms controlling implantation, namely the factors of immunity (antibodies, cells, cytokines and other mediators) whose exact regulation on the feto-maternal interface is a crucial precondition of successful implantation. Also the genetics of early embryo, as well as the possibilities of modern endoscopic techniques offer new insight onto mechanisms of implantation. Recommendations for diagnostics and treatment of implantation failure are given in the end of the article.

[European Cataract Outcome Study--results of five years of participation]. / European Cataract Outcome Study--výsledky petileté úcasti.

GOAL: To inform about European Cataract Outcome Study and to publish results from the Department of Ophthalmology, Faculty Hospital in Hradec Králové achieved during the five years' participation. METHOD, GROUPS: European Cataract Outcome Study is an international multicentric study pursuing quality of cataract surgery in individual departments of European countries. Its goal is to monitor the development of new trends and to be helpful in setting standards using the collected data. The Study evaluates results of all surgeries performed during one month. Our department participated in it in the years 1998-2002. In the first part of the trial the basic demographic data as well as the course of the surgeries were followed, in the second part results of examination during the first six months after the surgery were collected. RESULTS: Operations are mostly performed by means of the facoemulsification method in local anesthesia (subconjunctival or parabulbar), the stay in hospital varies in individual years between 35.1% and 56%, the percentage of implanted foldable (soft) intraocular lenses rises every year, in 2002 up to 61.5% (average of the whole study was 78.2% in 2002), the percentage of posterior lens capsule rupture during the procedure during the five year period in our department descends (range 3.7% to 1.2%). In the referred groups, 40.4% to 50.5% of eyes suffered from still another disease. The best corrected visual acuity 0.6 or better after the surgery achieved in individual years 64% to 78%. The deviation from planed refraction is between 0.69 to 0.88 diopters. The induced postoperative astigmatism varied in followed periods from 0.49 to 0.69 cylindrical diopters. CONCLUSION: The results of the study shows that the quality of surgeries performed in the department is on the high level and is fully comparable with contemporary European standard.

[Evidence based medicine and cost-effectiveness analysis in ophthalmology]. / Evidence based medicine, cost-effectiveness analýza v oftalmologii.

GOAL: To make the reader familiar with the term evidence based medicine (EBM), to explain the principle of cost-effectiveness analysis (price-profit), and to show its usefulness to compare the effectiveness of different medical procedures. METHOD: Based on few examples, in this article the relevance and calculation of important parameters of cost-effectiveness analysis (CE), as utility value (UV), quality adjusted life years (QALY) is explained. In addition, calculation of UV and QALY for the cataract surgery, including its complications, is provided. RESULTS: According to this method, laser photocoagulation and cryocoagulation of the early stages of retinopathy of prematurity, treatment of amblyopia, cataract surgery of one or both eyes, from the vitreoretinal procedures the early vitrectomy in cases of hemophtalmus in proliferative diabetic retinopathy or grid laser photocoagulation in diabetic macular edema or worsening of the visual acuity due to the branch retinal vein occlusion belong to highly effective procedures. On the other hand, to the procedures with low cost effectiveness belongs the treating of the central retinal artery occlusion with anterior chamber paracentesis, as well as with CO2 inhalation, or photodynamic therapy in choroidal neovascularization in age-related macular degeneration with visual acuity of the better eye 20/200. CONCLUSION: Cost-effectiveness analysis is a new perspective method evaluating successfulness of medical procedure comparing the final effect with the financial costs. In evaluation of effectiveness of individual procedures, three main aspects are considered: subjective feeling of influence of the disease on the patient's life, objective results of clinical examination and financial costs of the procedure. According to this method, the cataract surgery, as well as procedures in the pediatric ophthalmology belong to the most effective surgical methods.