Fatty acid acylated Fab-fragments of antibodies to neurospecific proteins as carriers for neuroleptic targeted delivery in brain

AbstractA method for targeted delivery of neuroleptics from blood in brain based on using Fab-fragments of antibodies to antigens of brain glia cells (acid gliofibrillar antigen and α2-glycoprotein) is suggested. The essence of the technique is that the molecule of neuroleptic (trifluoperazine) is conjugated with Fab-fragments of these antibodies. The conjugate thus obtained is modified by stearoylchloride in the system of Aerosol OT reversed micelles in octane. The study of the distribution of 125I-labelled conjugates in the rat organism after intracordial introduction is performed. On the contrary to the nonmodified conjugates and conjugate, containing fatty acylated Fab-fragments of antibodies, nonspecific to the rat brain, the conjugate of trifluoperazine with stearoylated Fab-fragments of antibodies to neurospecific antigens accumulate in brain tissues. The drastic increase of the neuroleptic activity of trifluoperazine resulting from its coupling with stearoylated Fab-fragments of antiglial antibodies is observed

Cytokines and neuron-specific proteins in pediatric viral encephalitis and convulsive syndrome. I. Viral encephalitis

Convulsive syndrome in children is manifested in the three forms: febrile convulsions in acute infections, symptomatic convulsions during acute neuroinfection, as well as onset of epilepsy requiring careful differentiation to prescribe adequate therapy. A threat of convulsive syndrome spreads beyond complications related to ongoing infection, because its development is associated with the risk of emerging symptomatic epilepsy in the future. Postencephalitic epilepsy developing in children within the first years after viral encephalitis has been specifically highlighted. A necessity to identify groups at risk of developing epilepsy gave a momentum to seek out for biomarkers of epileptogenesis reflecting the features of systemic and local inflammatory process in the central nervous system during the immune response to infection. Cytokines mainly mediating inflammation are currently examined being studied as candidate biomarkers of the risk of epilepsy. On the other hand, neuron-specific proteins known as inflammation biomarkers identified in various diseases of the central nervous system are being investigated to reveal brain cell injury in neuroinfections and epilepsy. Here we review publications assessing a potential to use inflammation biomarkers (cytokines and neuron-specific proteins) to diagnose and monitor pediatric neurological diseases associated with convulsive syndrome. The first part of the review describes the results of determining the inflammation biomarkers in the blood and cerebrospinal fluid during acute viral encephalitis/encephalopathy associated with various neurotropic viruses (herpes viruses, flaviviruses, enteroviruses). A significance of diverse biomarkers in predicting an outcome and long-term disease consequences are discussed

Subsets of cerebrospinal fluid lymphocytes in acute pediatric respiratory viral infection with meningeal syndrome

Current urgency of studying the intrathecal cellular immune response to infections of central nervous system is determined by limited knowledge on existing data about mechanisms of the brain immune protection in normal and diseased state. Implication of multi-colour flow cytometry in clinical laboratory diagnostics allowed to perform detailed studies of biological liquors, including cerebrospinal fluid (CSF). Currently, however, there are only scarce data on the lymphocyte subpopulations in CSF. Appropriate reference values remain a challenging issue. A study of CSF lymphocyte pool in absence of definite results at previous examination may be a potential way to resolve this problem. These clinical conditions include acute respiratory viral infections (ARVI), presenting with pseudomeningitidis (meningism) syndrome. The aim of this work was to characterize the subsets of lymphocytes from CSF of the children with ARVI with the meningism symptoms in order to get basic (control) values for diagnostics of inflammatory brain diseases. We have studied subpopulation composition of the CSF lymphocytes form in 27 children with ARVI complicated by the meningism (pseudomeningitidis) by means of flow cytometry using FACSCalibur analyzer with BD MultiTEST IMK Kit reagents. The data evaluation was performed with FlowJo software. We have studied relative contents of the main subsets, i.e., total Т cells (CD3+); Т helpers (CD3+CD4+Th); cytotoxic T cells (CD3+CD8+CTL); natural killers (СD3-CD16+CD56+NK); В cells (CD3-CD19+), and minor lymphocyte subpopulations: double-positive (DP) (CD3+CD4+CD8+); double-negative (DN) (CD3+CD4-CD8-) T cells; NKT (СD3+CD16+CD56+); CD3+CD8bright, CD3+CD8dim, CD3-CD8+NK. Statistical evaluation was carried out with standard GraphPad Prism 5 software. Among the main lymphocyte populations in CSF, T cell were predominant (96.2%), as well as their subpopulations, i.e., CD4Th (53.4%), and CD8+CTL (28.2%), with low amounts of NK (2.2%) and B cells (0.7%). The mean relative content of minor subpopulations (DN or DP T cells, and NKT cells) was, respectively, 5.3, 4.0, and 9%. Age dependence was revealed for the contents of major and minor lymphocyte subsets. With advancing age of the children, the relative numbers of CD3+ and CD4+Тh cells in CSF increase, as well as CD4/CD8 ratio, associated with decreased share of NK cells, like as DN and CD3+CD8dimТ cells. The results obtained are reflect some features of lymphocyte pool in CSF of the children without inflammatory process in CNS. Thus, they may be referred as control values (inflammation-free brain disorders) when studying immune pathogenesis of neuroinfections and other inflammatory diseases of CNS in the children from different age groups

Diseño e implementación de práctica para el Laboratorio de Comunicaciones 4

Describe una serie de prácticas de Laboratorio utilizando MATLAB y el dsPIC30F4013, para el Laboratorio de Comunicaciones 4 de la carrera de Ingeniería en Electrónic

Major and minor lymphocytes subpopulations in peripheral blood and cerebrospinal fluid of children with meningitis

Introduction. The analysis of current publications indicates at our insufficient understanding of subpopulation composition of lymphocytes in peripheral blood and cerebrospinal fluid (CSF) during pediatric neuroinfectious diseases. It has been found that the main lymphocyte populations are divided into many small (minor) subpopulations.The purpose of this research was to assess percentage of major and minor blood and CSF lymphocyte subsets in children with aseptic viral meningitis (AM) or bacterial purulent meningitis (BM).Materials and methods. Phenotyping of blood and CSF lymphocytes of children aged from 4 months to 17 years diagnosed with AM (n = 86) and BM (n = 39) was carried out by using flow cytometry. As a comparison group, we analyzed peripheral blood and CSF samples collected from children with acute respiratory viral infections (ARVIs) associated with syndrome of meningism (n = 27). There was evaluated percentage of the major cell subpopulations (CD3+ T-lymphocytes, T-helpers — CD3+CD4+ Th, cytotoxic T-lymphocytes — CD3+CD8+ CTL, natural killer cells — CD3-CD16+CD56+ NK, B-cells — CD3-CD19+), as well as minor lymphocyte subsets (double positive (DP) (CD3+CD4+CD8+), double negative (DN) (CD3+CD4-CD8-) T-cells, NKT (CD3+CD16+CD56+), CD3-CD8+ NK, CD3+CD8dim and CD3+CD8 8bright).Results. It was found that the acute period of BM and AM vs. the comparison group (ARVI) was characterized by significant differences in the blood and CSF composition of major and minor lymphocyte subsets. In particular, blood T-cells, Th, CTL, NK, NKT, DN, CD3-CD8+ NK, CD3+CD8bright and CD3+CD8dim dominated in parallel with significantly lowered B-cell frequency in AM vs. BM. In the CSF of children with AM, T-cells and Th prevailed, whereas count of B-cells and CD3-CD8+ NK was lower compared to those in BM. In addition, further differences were revealed in CSF and blood cell subset composition depending on nosological entity, while maintaining differences in some major and minor lymphocyte subpopulations lacked in the comparison group. Calculating the CSF/blood ratio for the major and minor lymphocyte subsets uncovered the prevalence for the majority of cell subpopulations (the coefficients ranged from 1.2 to 16.4) in the CSF of the comparison group (ARVI), except B-cells, NK and CD3-CD8+ NK (coefficients ranged from 0.07 to 0.31). AM and BM were featured with various changes in the CSF/blood ratio found for most of the studied subpopulations in the acute period as well as the recovery phase highlighted with characteristic traits for each nosological form.Conclusion. The data obtained indicate about finding specific features in the activation of systemic and intrathecal immune response during viral and bacterial meningitis in children, which may be used as an additional differential diagnostic criterion

Defect growth in multilayer chromium nitride/niobium nitride coatings produced by combined high power impulse magnetron sputtering and unbalance magnetron sputtering technique

In recent years, high power impulse magnetron sputtering (HIPIMS) has caught the attention of users due to its ability to produce dense coatings. However, microscopic studies have shown that HIPIMS deposited coatings can suffer from some surface imperfections even though the overall number of defects can be significantly lower compared to, for example, arc deposited coatings of similar thicknesses. Defects can degrade the coating performance thus any kind of defect is undesirable. To better understand the nature of these imperfections and the science of their formation, a series of Chromium Nitride/Niobium Nitride (CrN/NbN) coatings were deposited using HIPIMS technique combined with unbalanced magnetron sputtering (UBM) by varying deposition times (t = 15 to 120 minutes). All other deposition parameters were kept constant in order to deposit these coatings with a consistent deposition rate and stoichiometry. In addition, coatings were deposited using pure UBM technique to compare the defects generated by these two different physical vapour deposition approaches. High-resolution scanning electron microscopy images revealed that HIPIMS/UBM and pure UBM CrN/NbN coatings have similar types of defects which could be categorised as: nodular, open void, cone-like and pinhole. Interestingly, there was no evidence of droplet formation in HIPIMS/UBM deposited coatings. The defect density calculation indicated that the defect density of HIPIMS/UBM coatings increased (from 0.48 to 3.18%) with the coating thickness. A coating produced in a relatively clean chamber had a lower defect density. Potentiodynamic polarisation experiments showed that the fluctuation in corrosion currents in HIPIMS/UBM coatings reduced with the coating thickness. This indicated that though visible on the surface, most of these defects did not penetrate thorough the whole thickness of the coating