A new set of reagents and related software used for NGS based classical and non-classical HLA typing showing evidence for a greater HLA haplotype diversity.

To evaluate the HLA typing performance of a new Long-Range PCR NGS set of reagents and its dedicated software, a panel of 41 reference homozygous cell lines from the International Histocompatibility Working Group (IHWG) and a panel of 376 volunteer bone marrow donors were analyzed for classical and non-classical HLA class I and class II genes. All results, except HLA-DPB1, were obtained without any ambiguities at the 3rd field level. Based on the high resolution performance of the reagents, a number of new alleles have been described not only for classical but also for non-classical HLA class I genes, leading to a more accurate haplotype definition. Linkage disequilibrium between HLA-A and HLA-G genes has been defined at 4th field level of resolution. Moreover, for the first time, HLA-DQA2 and DQB2 polymorphisms and their linkage disequilibrium with DQB1 were described.

Evidence for a higher resolution of HLA genotyping by a new NGS-based approach.

With more than 16,000 alleles identified, the human leucocyte antigen (HLA) system is one of the most polymorphic regions of the human genome. Regarding the crucial role of HLA compatibility in transplantation and especially in Hematopoietic Stem Cell Transplantation, identification of HLA polymorphisms at a high-resolution level is of major interest. Recently, NGS technology has been proposed which appears to be simpler and more informative than the classical molecular methods such as SSP, SSOr and SBT. In the present report, a new set of NGS reagents and the appropriate associated software for sequence analysis are described. Through different studies, the performances of the system are illustrated and demonstrate that the method herein described overcomes current limitations in performing high-resolution HLA typing in clinical laboratories.

Radon in groundwater and dose estimation for inhabitants in Spas of the Sudety Mountain area, Poland.

Studies of radon isotope (222)Rn activity concentration in underground water in the Sudety region were performed with the use of the liquid scintillation technique. Waters chosen for investigations were collected in 24 health resorts and towns of the Sudety area from 115 springs, wells and intakes. The (222)Rn activity concentration varied within the range from 4.2+/-0.4 to 1703+/-55 Bq/l. The annual effective doses due to the consumption of (222)Rn with water were calculated for 50 sources of underground spring water or tap water used for consumption. The results were within the range from 0.003 to 1.1 mSv/yr, assuming 0.5 l of tap water per day from which radon is not removed or 0.5 l of mineral spring water consumed daily. The contribution to the effective dose from the inhalation of radon during the daily usage of domestic water substantially increases its effective dose.

Natural radioactivity in underground water from the Outer Carpathians in Poland with the use of nuclear spectrometry techniques.

The investigations of natural radioactivity in underground mineral water and spring water in health resorts in the Outer Carpathians were performed. Samples from 40 water springs were collected 3-4 times over a period of 10 years (1997-2007). In order to obtain necessary data, two different nuclear spectrometry techniques were applied: WinSpectral alphabeta 1414 liquid scintillation counter from Wallac and alpha-spectrometer 7401 VR from Canberra-Packard, USA with the silicon surface barrier detector. The activity concentrations of (222)Rn in the investigated samples varied from below 1 to 50 Bq/l. For radium isotopes the concentrations were in a range from below 10 to 490 mBq/l for (226)Ra and from 29 to 397 mBq/l for (228)Ra. The highest concentrations for both radium isotopes were obtained for medicinal water Zuber III from Krynica spa. The activity concentrations for uranium isotopes varied from below 0.5 to 16 mBq/l for (238)U and from below 0.5 to 162 mBq/l for (234)U with the highest values obtained for water Zuber IV. The highest annual effective dose arising from mainly radium isotopes was obtained for Zuber III water and was equal to 75 microSv/yr. Additionally, the annual effective doses due to (222)Rn consumed with water were also estimated. The isotopic ratios between isotopes originating from the same decay chain ((234)U/(238)U, (226)Ra/(238)U) and from different radioactive decay chains ((226)Ra/(228)Ra) were determined. The correlations between different isotopes were presented.

Natural radioactivity and dose estimation in underground water from the Sudety Mountains in Poland.

Studies of natural radioactivity in spring water in Swieradów-Czerniawa Health Resort were performed with the use of nuclear and mass spectrometry techniques. The investigated region is situated in the Sudety Mountains, where uranium exploration was conducted in the early 1950s of the last century. Annual effective doses due to radionuclide intake were calculated for 4 out of 20 spring waters used for consumption by spa patients and inhabitants. The summed effective doses without 222Rn were of the range 0.4 microSv to 9.2 microSv, for patient for of a 20-day duration stay and from 1.3 microSv y(-1) to 26.7 microSv y(-1) for an inhabitant. The contribution of radon consumed with water raises these values to 209.4 microSv per 20 days and 608.3 microSv y(-1) for a patient and inhabitant, respectively.

Cerebrospinal fluid pulse pressure waveform analysis in hydrocephalic children.

Analysis of cerebrospinal fluid pulse pressure was reported to be useful in the assessment of the cerebrospinal pressure-volume compensation. The method for the estimation of high-frequency centroid (HFC) was modified and used to verify the correlation between HFC and other compensatory parameters investigated by means of the lumbar infusion test in 94 hydrocephalic children. The results confirm that in hydrocephalus HFC is positively correlated to cerebrospinal elasticity coefficient, but inversely to mean CSF pressure and pulse wave amplitude. It was also demonstrated that HFC decreased dynamically during the infusion test.

Identification of the cerebrospinal compensatory mechanisms via computer-controlled drainage of the cerebrospinal fluid.

The constant rate infusion test is still widely used for the diagnosis of hydrocephalus in children. The increased resistance to reabsorption of the cerebrospinal fluid (CSF) is considered the most important factor in the improvement seen after shunting. However, the classic infusion test has some disadvantages: the compensatory model is identified in the conditions of raised intracranial pressure, which may provoke some uncontrolled changes in cerebral blood volume in the mechanism of vasomotor response; also non-linear effects in reabsorption mechanisms may have a significant influence. The controlled drainage of CSF can be considered as a form of pressure-volume testing if the volume of the drained fluid is precisely measured. The main advantage of the method is the possibility of estimating unknown parameters in practically physiological conditions (with only slightly decreased pressure). This paper presents a new method for the identification of an electrical model of cerebrospinal volume compensation. The method has been described theoretically, verified on the simulator and introduced into clinical practice at the Medical Academy in Warsaw and the Children's Health Center in Miedzylesie.

Cerebrospinal compensation in hydrocephalic children.

One hundred and fifteen cases of hydrocephalus in children were analysed. Cerebrospinal compensatory reserve was assessed by a computerized, constant rate, lumbar infusion test. Head circumference and ventricular size were measured and a psychometric examination carried out. A classification of hydrocephalus based on resting cerebrospinal fluid pressure (CSFP) and resistance to cerebrospinal fluid outflow (RCSF) was introduced. Parameters of compensatory reserve were compared in atrophy (low CSFP, low RCSF), normal-pressure hydrocephalus (low CSFP, increased RCSF), non-communicating hydrocephalus (high CSFP, low RCSF) and acute hydrocephalus (high CSFP and increased RCSF). Significant differences were found between the factors describing compensatory ability in these groups. Sixty-two patients could be classified on the basis of resting CSFP and RCSF. Differentiation between the types of hydrocephalus was shown to be more accurate when all variables measured during the pressure-volume test were considered. The patterns of the time courses of CSFP during rate infusion tests in the different types of hydrocephalus are presented.

[Post-traumatic intracerebral hematoma]. / Pourazowe krwiaki sródmózgowe.

Fifty-five cases of post-traumatic intracerebral haematomas are analyzed, discussing the diagnostic value of such preliminary methods as plain skull films, EEG, echoencephalography. These methods, together with clinical findings make the diagnosis of intracerebral haematoma likely. The final diagnosis was based on carotid arteriography and computerized tomography of the brain, which provided additional information on traumatic brain damage. The diagnostic sensitivity of CT was higher. The considerable prognostic importance of the degree of consciousness disturbances and their duration is stressed. In the group of patients with lucidum intervallum the mortality was higher. Four patients were treated conservatively since CT demonstrated in them only small haematomas without displacement of the ventricles. The remaining patients were treated surgically removing the haematomas through craniotomy or craniectomy. In 54% of these cases improvement was obtained. The 33% mortality was moderate as compared with previous reports.