[Neurological complications and outcomes of bacterial meningitis in children]. / Nevrologicheskie oslozhneniya i iskhody bakterial'nykh meningitov u detei.

AIM: To analyze neurological complications and outcomes of bacterial meningitis (BM) in children. MATERIAL AND METHODS: Fifty-five patients with BM, aged from 2 months to 12 years, were examined. Bacteriological study, clinical and biochemical blood tests and blood serology and cerebrospinal fluid (CSF) tests as well as serum molecular-genetic study were performed. Neuroimaging methods (neurosonography, computed (X-ray) tomography and brain magnetic resonance imaging) were used. RESULTS AND СONCLUSION: A key role of generalized meningococcal infection in the development of BM in children was confirmed. Brain edema was an early and life-threatening complication of BM. It was found in 9% of the patients with meningococcal infection, 7.3% with pneumococcal meningitis and 3.6% with haemophilus meningitis. Changes in the brain structure were not found in 80% of the patients, in 20%, the residual stage of BM was characterized by cerebral destructive/proliferative or atrophic changes of different severity. The dissociation between clinical and neuroimaging parameters and poor outcomes of BM, related to the complicated premorbid state, concomitant somatic/neurological pathology, BM severity, late laboratory diagnosis and untimely etiotropic antibacterial treatment, were identified in 65.5%.

[Neurological presentations of bacterial meningitis in children: current possibilities of diagnosis and treatment]. / Nevrologicheskie proyavleniya bakterial'nykh meningitov u detei: sovremennye vozmozhnosti diagnostiki i lecheniya.

AIM: To improve clinical/laboratory diagnosis of bacterial meningitis (BM) in children. MATERIAL AND METHODS: Fifty-seven patients, aged from 4 months to 12 years, with BM were examined. Bacteriological study of the mucus from the nasopharynx, sterility tests of blood and cerebrospinal fluid (CNF), clinical and biochemical tests, serological study of paired samples of plasma - indirect hemagglutination test, latex agglutination assay, immunological study of plasma and CNF with the determination of interferon -α, -ß and -γ, interferon procalcitonin and neopterin with ELISA were performed at admission and after CNF remediation. Molecular-genetic study of the blood serum and CNF using PCR, neurosonography, computed tomography and MRI of the brain were done as well. RESULTS AND CONCLUSION: Most of the patients had generalized meningococcus infection (64,9%). Hib-meningitis, pneumococcus meningitis and other forms were less frequent. Characteristics of neurological presentations of BM depending on the etiology, somatic complications and time from the manifestation of symptoms were described. Clinical/biochemical criteria of BM and correlations between procalcitonin, neopterin and interleukin-8 in the CNF and blood of the patients and disease severity as well as between the level of inflammatory reactions in the CNF and the BM form were established. These results are important for the development of informative biomarkers for the prognosis of BM course and outcome.

[Preimplantation genetic diagnosis and monogenic inherited eye diseases]. / Preimplantacní genetická diagnostika a dedicná onemocnení oka.

OBJECTIVE: Preimplantation genetic diagnosis (PGD) is an established application of genetic testing in the context of in vitro fertilization. PGD is an alternative method to prenatal diagnosis which aims to prevent the transmission of an inherited disorder to the progeny by implanting only embryos that do not carry genetic predisposition for a particular disease. The aim of this study is to provide an overview of eye disorders for which PGD has been carried out. METHODS: The European literature search focused on best practices, ethical issues, risks and results of PGD for inherited eye disorders. RESULTS: PGD is performed for a number of ocular disorders; a prerequisite for its application is however, the knowledge of a disease-causing mutation(s). The main advantage of this method is that the couple is not exposed to a decision of whether or not to undergo an abortion. Qualified counselling must be provided prior to the PGD in order to completely understand the risk of disability in any child conceived, consequences of disease manifestation, and advantages as well as limitations of this method. In the group of non-syndromic eye diseases and diseases in which ocular findings dominate, PGD has been performed in European countries for aniridia, choroideremia, congenital fibrosis of extraocular muscles, Leber congenital amaurosis, ocular albinism, retinitis pigmentosa, X-linked retinoschisis, Stargardt disease, blepharophimosis-ptosis-inverse epicanthus syndrome and retinoblastoma. Sexing for X-linked or mitochondrial diseases has been carried out for blue cone monochromatism, choroideremia, familial exudative vitreoretinopathy, Leber hereditary optic neuropathy, macular dystrophy (not further specified), Norrie disease, X-linked congenital stationary night blindness, X-linked retinoschisis and nystagmus (not further specified). CONCLUSION: In recent years, there has been an increase in potential to use PGD. The spectrum of diseases for this method has widened to include severe inherited eye diseases.Key words: preimplantation genetic diagnosis; monogenic eye diseases; in vitro fertilization.

Preimplantation genetic diagnosis of X-linked diseases examined by indirect linkage analysis.

BACKGROUND: Many centers of assisted reproduction in the Czech Republic offer preimplantation genetic diagnosis with fluorescent in situ hybridization (FISH) to couples requiring preimplantation genetic diagnosis (PGD) of X-linked diseases. However, this process results in discarding all male embryos and is not able to distinguish a carrier or healthy female embryo in X-linked recessive disorders. OBJECTIVES: The main aim of this study was to summarize a six-year period of PGD of X-linked monogenic diseases using indirect linkage analysis. METHODS AND RESULTS: We wanted to accentuate the advantage indirect analysis of PGD using multiple displacement amplification (MDA) followed by short tandem repeat (STR) analysis. We present forty-six PGD cycles, including pre-case haplotyping (PGH) panel, for fifteen X-linked diseases. Embryo transfer was made thirty-eight times and gravidity was confirmed in thirteen female probands with a success rate of pregnancy calculated at 42 %. CONCLUSIONS: PGD procedure using MDA amplification followed by STR analysis provides help in identifying genetic defects within embryos prior to implantation. The reliability of the method was also supported by high pregnancy rate compared to other publications, which commonly achieved a 30-35 % success rate (Tab. 2, Fig. 1, Ref. 33).

[Implementation of arrays in first trimester prenatal diagnosis]. / Implementace array vysetrení do prenatální diagnostiky v I. trimestru.

OBJECTIVE: Array technology in chorionic villus sampling (CVS) - analysis of clinical benefit and a proposal of a more effective 1st trimester genetic testing policy. DESIGN: Retrospective study. SETTING: Gennet, Center of Medical Genetics and Reproductive Medicine, Prague. MATERIAL AND METHODS: Total of 913 CVS were performed at Gennet between 2010-2014. All 913 samples were tested by QF-PCR rapid test for aneuploidy of chromosomes 13, 18, 21, X and Y and karyotyping following standard long term culture. Microarray analysis (Illumina HumanCytoSNP12 v2.1) was performed on 179 samples with normal result from both - QF-PCR and karyotyping. RESULTS: At 229 samples the common chromosomal aneuploidy was detected using rapid QF-PCR (25% from 911 successful rapid tests). Conventional karyotyping revealed 239 unbalanced chromosome aberrations (27% from 897 successful cultivations). 227/239 (95%) positive karyotypes confirmed QF-PCR finding of common aneuploidies. 10 unbalanced chromosome aberrations were not covered by rapid QF-PCR test. Microarray analysis of samples with normal result from both- QF-PCR and karyotyping- revealed 13 clinically relevant chromosome aberrations (7.5%). CONCLUSION: New policy for chorionic villi testing at Gennet was established. Based on evaluation of the results of karyotyping, array and QF-PCR and analysis of published data we decided to replace karyotyping by microarray analysis in all cases of foetuses with normal results from QF-PCR. More effective detection of pathological and clinically relevant chromosome aberrations in examined foetuses is expected.

[Palivizumab: four seasons in Russia].

In 2010, the Russian Federation (RF) registered palivizumab--innovative drug, based on monoclonal antibodies for passive immunization of seasonal respiratory syncytial virus (RSV) infection in children of disease severe progress risk group, which include primarily premature infants, children with bronchopulmonary dysplasia and hemodynamically significant congenital heart disease. Currently, palivizumab is included in the list of recommended medicines and medical care standards of different countries, including Russia. In the review the results of Russian research on the progress of RSV infection, its epidemiology and immunization experience gained over the 2010-2014 period are summarized in relation to the foreign data. During the four epidemic seasons palivizumab immunization covered more than 3,200 children of severe RSV infection risk group with a progressive annual increase in the number of patients who received the drug. Geography of palivizumab immunization is also greatly expanded in our country during this time. If during the first two seasons measures of immunization were taken mainly in Moscow and St. Petersburg, at the present time, thirty one territorial entities of the Russian Federation have the experience in the drug application. Analysis of the results of RSV infection immunization (made in several regions) confirms the high clinical efficacy and palivizumab safety already demonstrated in international studies. In addition, the analysis presents the potential to improve the efficiency of the integrated RSV infection immunization programs, realizing in the establishment of high-risk child group register, adequate counseling for parents, as well as the development of the routing of patients and coordination of interaction between different health institutions during the immunization.

[Application of SNP array method in prenatal diagnosis]. / Aplikace metody SNP array v prenatální diagnostice.

OBJECTIVES: SNP array (array method using Single Nucleotide Polymorphisms) enables to detect cytogenetically undetectable submicroscopic alterations (microdeletions, microduplications), which could be also causative for ultrasonographic anomalies of fetus. This article describes the principle, advantages, disadvantages and application possibilities of the SNP array method in prenatal diagnosis. The ten month experience with SNP array use in prenatal diagnosis is presented. DESIGN: Prospective study. SETTINGS: Gennet, Prague. MATERIAL AND METHODS: During the period from April 2010 to January 2011 we performed 110 SNP array analyses of fetal DNA: 14 chorionic villi samples (CVS), 88 amniotic fluid samples (AMC), 1 cord blood sample and 7 miscarriage samples. Laboratory tests were carried out on DNA from both cultured and uncultured fetal cells. Examinations were performed in fetuses with sonographic abnormal findings having normal karyotype. In addition 14 fetal cytogenetic abnormalities were solved. SNP array analysis was performed using Illumina InfiniumHD HumanCytoSNP-12 chip. All data were analysed by Illumina KaryoStudio and GenomeStudio software. RESULTS: SNP array analysis was performed in 108 fetuses (only 2 examination failures, 1.8%). In total, we detected CNV (copy number variation) in 29 samples (29/108 = 27%). 15% (16/108) of fetuses with abnormal ultrasound findings were found to carry clinically relevant CNV. Probably benign CNVs were found in 8 samples (8/108 = 7%) and in additional 5 CNVs parental samples have not been analysed yet. Excluding karyotypically abnormal cases clinically relevant CNVs were found in 10% of fetuses (9/94). In all cases with de novo chromosomal aberration the clinical relevancy was clarified (imbalances in 50%). CONCLUSION: Our data suggest that SNP array analysis is a relevant and useful technique in prenatal diagnosis.

[Impact of herpesvirus infections on the level of proinflammatory cytokines in premature neonatal infants].

The aim of this study was to estimate the frequency of HSV and/or CMV among the pathogens causing intrauterine infections (IUI) and to investigate their impact on the level of proinflammatory cytokines in premature neonatal infants. Examinations were performed in 3 neonatal groups: 1) premature neonates with clinical manifestations of IUI; 2) those without IUI; 3) full-term newborns. In group 1, viral (HSV and/or CMV) and bacterial infections were detectable with the same frequency. Quantitative analysis of plasma IL-6 and IL-8 levels and the induced production of these cytokines by blood cells in vitro showed that in Group 1 neonates, IL-6 and IL-8 concentrations were substantially higher and the induced production of these cytokines was lowerthan those in Group 3. The detection of HSV and/or CMV markers in premature newborn infants was attended by a statistically significant rise in plasma IL-6 levels; the identification of the opportunistic bacterial microflora correlated with the higher concentration of IL-8. In Group 1, wiferon produced an immunomodulatory effect, by lowering IL-8 concentrations to the level observed in Group 3.

[A quantitative analysis of the development of the auditory centers of the medulla oblongata in the hen]. / Kolichestvennyi analiz razvitiia slukhovykh tsentrov prodolgovatogo mozga kuritsy.

Development of three auditory nuclei in medulla oblongata was studied in the chicken (Gallus gallus). Quantitative description of the volume growth of the magnocellular nucleus neurons and linear growth of the auditory nuclei is given from the formation of differentiated neuroblasts (Day 9 of embryogenesis) until the 180th day of postembryonic development. Growth of the embryos was described according to the Bertalanffy's model. Growth of the neurons proceeded at the highest rate at the rostral level of magnocellular nucleus (mN = 1.204 microns/day) and the lowest rate of the caudal level (mN = 1.045 microns/day). The curve of linear growth of the auditory nuclei is similar with the growth curve of the neurons and follows the Bertalanffy's equations. The magnocellular nucleus is characterized by the fastest growth (mN = 75.6 microns/day) and the angular nucleus by the slowest growth (mN = 52.0 microns/day). The growth of the neurons and auditory nuclei decreases before hatching.

[Ionol phosphorus organic analogs and stabilization of levorin]. / Ispol'zovaniephosphoroorganicheskikh analogov ionola dlia stabilizatsii levorina.

Stability of levorin isolated and purified with the use of ionole phosphorus organic analogs having in their structure phosphate, phosphonate and phosphonite groups was studied. The compound having in its structure (in addition to tertiary butyl substitutes) the phosphonite group with the P-H bond increasing the compound antioxidant property was shown to have the highest stabilizing effect. The results of the study made it possible to recommend the use of the space complicated phenols with the structure fragments of the P-H bond type as antioxidants in production of levorin.

[Water-soluble phosphorus-containing forms of ionol in experimental treatment of thermal burns]. / Vodorastvorimye fosforsoderzhashchie formy ionola v éksperimental'noi terapii termicheskikh ozhogov.

The authors studied the membrane-stabilizing effect of two water-soluble preparations of antioxidants, analogues of ionol, in thermal burns. It was found that both preparations increase somewhat to a different degree the resistance of the red cell membranes to peroxide compounds and reduce hemolysis which is very characteristic of the first minutes and hours after the thermal trauma. The preparations alter the electrical properties of the red cells and affect the activity of phospholipases in blood plasma.

[Monoclonal antibodies to alpha-endorphin effective in immunohistochemistry and immunoblotting]. / Monoklonal'nye antitela k al'fa-éndorfinu, éffektivnye v immunogistokhimii i immunoblottinge.

Four mouse monoclonal antibodies (E11, A8, F8, H5) to alpha-endorphin have been produced. The antibodies bind 12.5, 20.6, 9.6, 6.6% of 125I-beta-endorphin and 35.5, 15.1, 12.8, 12.2% of 125I-gamma-endorphin; the binding of 125I-alpha-endorphin being taken for 100%. The binding of antibodies E11, A8, F8 and H5 to 125I-alpha-endorphin was 50% inhibited by unlabeled ligand in concentrations 5, 50, 30 and 35 nM respectively. Using tissue sections of rat pituitary it was shown that antibody E11 can be used for the localization of endorphin producing cells by immunofluorescence. The antibodies F8 and H5 effectively detected endorphin precursor proopiomelanocortin by immunoblotting.