Development of a recombinant NP protein based indirect ELISA for the detection of antibodies against Newcastle disease virus and differentiation of infected or recombinant vaccine immunized chickens.

Newcastle disease (ND) is a highly contagious and economically important disease in the poultry industry caused by avian avulavirus-1, historically known as Newcastle disease virus (NDV). Control of ND primarily relies on prophylactic vaccination of flocks, and many vaccines are available on the market, both conventional and more recently introduced new generation recombinant types. To assess the protection level achieved by vaccination ELISA tests are typically used, they also are to track an infection with field strains in non-vaccinated flocks. Special modifications of ELISA can be used as a screening tool to detect infection in flocks vaccinated with new generation vaccines. In this study, we have developed an ELISA test for the detection of antibodies against the nucleoprotein (NP) of NDV and for differentiation of chickens vaccinated with commercial and prototype in-house recombinant vector vaccines from those infected with field NDV strains. The NP gene of LaSota NDV strain expressed in a baculovirus vector was used as a coating antigen in the ELISA. The developed test was optimized, validated and compared to other serological tests. The sensitivity, specificity and accuracy of recombinant NP protein-based ELISA were respectively 96.1%, 96.3%, and 96.2%. Inter-rater (kappa) agreement between the NP-ELISA and the gold standard HI test was calculated to be 0.995. In our comparisons, commercially available ELISA tests revealed different specificities ranging from 95.5-100% and sensitivities at variance, ranging from 90.1 to 99.0%. A high level of maternally derived antibodies was measured in the serum of 1-day-old broilers in the NP-ELISA assay. These antibodies had disappeared and were undetected at 3, 5 and 6 weeks post-vaccination but birds became positive again at 2 weeks after control infection with a velogenic NDV strain. In SPF chickens, antibodies against NP protein were detected only after a challenge. The recombinant NP protein-based ELISA test is sensitive, specific and accurate when compared to the gold standard HI test and commercially available kits. Moreover, the method could be also used for the differentiation between vaccinated and infected birds.

Analysis of corpus callosum size depending on age and sex.

BACKGROUND: The aim of the study was to analyse changes in the size of the corpus callosum (CC) depending on age and sex and to establish the reference values of the morphometric indices of the CC in the Polish population. MATERIALS AND METHODS: The results of magnetic resonance studies of 1108 patients performed in the years 2010-2014 were analysed. Two independent radiologists evaluated cerebral images to exclude deviations from normal state. In patients divided according to sex and to 10 age groups, measurements of CC and brain dimensions were made and morphometric indices were calculated. RESULTS: The results of measurements related to the following parameters: lengths of longitudinal cross-section of CC (CD), CC thickness in the narrowest place - isthmus (EF), the largest linear dimension of the brain from the frontal pole to the occipital pole (AB), the longitudinal cross-section area of the CC (A1) and cerebral cross-section area (A2) as well as CD/AB and A1/A2 ratios are summarised in 7 figures and 3 tables. CONCLUSIONS: It was demonstrated, that in all age groups there are statistically significant differences in the values of the analysed parameters and ratios of CC size. It was indicated, that there are no statistically significant differences between men and women in the CD, EF, and A1 parameters related to CC size, and the profiles of variations of these parameters are very similar. It was proved that the- re are statistical differences between women and men in parameters/indicators concerning of the brain size.

Specific detection of GII-1 lineage of infectious bronchitis virus.

Infectious bronchitis virus (IBV) is a worldwide prevalent RNA virus that causes highly contagious and economically devastating disease in chicken. The virus exists in many different genetic forms which made the disease control very difficult. The present study describes the development and validation of TaqMan probe-based real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) targeting the S1 coding region of S gene characteristic for the GII-1 lineage (formerly the D1466-like variant) of IBV. These strains are quite different from other European IBV belonging to different lineages of the GI genotype. The developed method was 30-fold more sensitive than used so far for standard nested RT-PCR with detection limit of 56 RNA copies per reaction. The specificity of the assay was also evaluated with a panel of different poultry pathogens. Repeatability and reproducibility of the method was very high with coefficients of variation lower than 4%. One hundred and twenty-seven IBV-positive samples were tested by this method and GII-1 strains were detected in four of them (3·15%) which indicate a decrease in the GII-1 IBV prevalence in Poland. The assay was proven to be a valuable tool for rapid diagnosis of GII-1 lineage of IBV strains and moreover it enabled the monitoring of viral loads which can be used to assess disease progression. SIGNIFICANCE AND IMPACT OF THE STUDY: This study reports a TaqMan probe-based real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) for rapid and accurate identification of GII-1 lineage (formerly D1466-like variant) of infectious bronchitis virus (IBV). The assay revealed to be more sensitive than standard nested RT-PCR assay, previously used for this purpose. The developed assay has been tested on numerous field samples and revealed 3·15% prevalence of this lineage of IBV in Polish chicken population. Moreover, this new assay enables the assessment of viral load measurement which might be useful for epidemiology and pathogenesis studies.

[Spasticity and physical methods for controlling it]. / Spastycznosc i metody fizykalne jej zwalczania.

Spasticity is one of the greatest difficulties in patients with central nervous system injuries and diseases. Severe spasticity makes treatment, rehabilitation and care of patient very difficult and sometimes even impossible. It has been sought for many years for an objective method to evaluate the degree of spasticity, necessary to establish the results of treatment and rehabilitation. In this study we present subjective and objective methods of evaluating the spasticity in order to classify every patient to adequate therapeutic group. The authors present physical methods that not only contribute to control of spasticity together with pharmacotherapy and surgical treatment, but can be used alone. The big advantage of this therapy is a low invasiveness and the very few side effects.