Increasing signal-to-noise ratio in over-determined Mueller matrices.

This work investigates how the signal-to-noise ratio (SNR) of an over-determined Mueller matrix can be improved by changing the method of calculation. Specifically, our investigation focused on comparing SNRs achieved using the vector methodology from the field of partial Mueller polarimetry, and the matrix methodology. We use experimentally derived measurements from an investigation into the time-varying signal produced by the Mueller matrix of an electro-optic Bismuth Silicon Oxide (BSO) crystal undergoing cyclical impact of a Helium plasma ionisation wave. Our findings show that the vector methodology is superior to the matrix methodology, with a maximum SNR of 7.54 versus 4.97. We put forth that the superiority of the vector methodology is due to its greater flexibility, which results in the Mueller matrix being calculated with better condition matrices, and higher levels of SNR in the intensity measurements used for calculation.

Optimizing Mueller polarimetry in noisy systems through over-determination.

Mueller polarimetry measurements are increasingly being used to image highly dynamic and short-lived phenomena such as plasma discharges. For phenomena such as these, exposure times below 1 µs must be used. Unfortunately, these low exposure times significantly reduce the signal-to-noise ratio, making accurate and consistent measurements difficult. To overcome this limitation, we investigated increasing the number of Stokes vectors produced from a polarization state analyzer and polarization state generator, a process known as over-determination. To conduct our analysis, we used results from physical experiments using Stokes vectors generated by liquid crystal variable retarders. These results were then verified using data from simulations. First, we conclude that increasing the degree of over-determination is a simple and effective way of dealing with this noise; however, we also convey that choosing the best scheme is not an entirely trivial process. Second, we demonstrate that over-determination gives rise to hitherto inaccessible information that allows for the quantification of statistical noise and, crucially, the pinpointing of the origin of systematic error, a highly beneficial process that has been lacking until now.

Effect of blend moisture and extrusion temperature on physical properties of everlasting pea-wheat extrudates.

The effect of everlasting pea in combination with wheat on physical properties and microstructure of extrudates were studied. The share of everlasting pea (Lathyrus sativus) was variable, at 35, 50 and 65 %, respectively. The everlasting pea-wheat mixtures were moistened to the required level (18, 21, and 24 %), homogenized, conditioned and extruded in twin-screw extruder with counter-rotating conical screws. All of the obtained extrudates were characterised by a slow degree of radial expansion and high specific density. The Pearson correlation analysis indicated a statistically significant linear Pearson correlation (p < 0.05) between chemical compositions of the blends and physical properties of the extrudates. The expansion ratio increased as the concentration of the fibers and proteins increased, while specific density and hardness decreased. Inverse relationship was observed for crude fat. The microstructure of the extrudates was determined by both the moisture of the blend and the process temperature. The differences observed in the size, number of air cells and in the cell wall shapes and thickness indicate possibilities of the modification of physical properties of everlasting pea-wheat extrudates. The extrudates produced from everlasting pea-wheat blends (50:50) at higher barrel temperature (110/140/180/170/130 °C) were characterised by more numerous air cells of smaller diameters. Increasing moisture content of extruded blends results in extrudates with a higher porosity. No significant effect was shown in the chemical compositions on the level of metal contamination in the extrudates. The application of a counter-rotating twin-screw extrusion-cooker in the study permitted the production of compact, hard everlasting pea-wheat extrudates for use in vegetarian lunch dishes.

Safety of intravenous thrombolysis for acute ischaemic stroke including concomitant neoplastic disease sufferers - experience from Poland.

BACKGROUND: Ischaemic stroke (IS), brain haemorrhage and cerebral venous thrombosis can occur as an early and late complication of cancer in the clinical course. Cancer patients are at increased risk for stroke from direct and indirect effects of their malignancy. AIMS: The aim of our study was to evaluate the relationship between neoplastic disease and the long-term outcome, mortality and the presence of haemorrhagic complications in patients with acute IS treated with i.v. thrombolysis. METHODS: We retrospectively evaluated the demographic and clinical data of 495 Caucasian patients with acute IS and 40 patients with IS and concomitant neoplastic disease who were consecutively treated from 2006 to 2013 in two experienced stroke centres. RESULTS: In analysed group, there were 7.8% of patients with cancer [50.0% male, mean age 72.3 ± 9.3; National Institutes of Health Stroke Scale - 13 (range 9.5-17)]. Cancer was diagnosed before i.v.-thrombolysis in 28 (70.0%) patients. After 3 months of follow up, 60% of patients were independent (mRS 0-2) compared with the group of patients without cancer - 55% (p = 0.54), 17.5% died (18.4%; p = 0.89), 12.4% suffered haemorrhagic transformation (HT) (17.6%; p = 0.41) and 2.5% experienced SICH (4.4%; p = 0.56, respectively). Other clinical complications were not found. A multivariate analysis showed no impact of neoplastic disease on unfavourable outcomes [modified Rankin scale 3-6)] after 3 months (p = 0.15). CONCLUSION: Intravenous thrombolysis performed in Caucasian stroke patients with past or current neoplastic diseases, but not in the course of chemo- and radiotherapy, can be a safe and effective method of treatment. In making decision on the thrombolytic treatment, the risk of bleeding complications and the life expectancy should be assessed.

The efficacy and safety of intravenous thrombolysis with alteplase in the treatment of ischaemic stroke in a rural hospital.

BACKGROUND AND PURPOSE: Ongoing evaluation of the results of thrombolytic therapy in patients with ischaemic stroke (IS) in regions with different health care organization is absolutely crucial for making this method of treatment safer and efficient. The aim of this study was to analyse the efficacy and safety of treatment with intravenous alteplase in patients with acute IS in a rural hospital. MATERIAL AND METHODS: Between 2006 and 2011, 1392 pa-tients with IS were treated (including 200 patients treated with alteplase; 14.37%). In patients treated with alteplase, we analysed the influence of several variables on the functional status after 3 months according to the modified Rankin Scale (mRS), case-fatality rate during 3 months after onset and symptomatic intracerebral haemorrhage (SICH). RESULTS: In the studied population, good outcome (mRS 0-2) at 3 months was related to younger age (p = 0.001), male sex (p = 0.02) and low scores (< 15 points) on the National Institutes of Health Stroke Scale (NIHSS) (p < 0.0001). Deaths within 3 months were related to older age (p = 0.027), female sex (p = 0.004), severity of stroke measured by NIHSS score (p < 0.0001) and presence of radiological signs of previous stroke in baseline computed tomography (CT)(p = 0.002). Patients with SICH had higher mean age (p = 0.014) and higher severity of neurological deficit measured on the NIHSS scale (p = 0.03). CONCLUSIONS: The indications for intravenous thrombolysis in patients with IS should be strictly analysed so that the treatment is effective and safe especially in older patients, patients with greater severity of neurological symptoms and patients with old post-stroke lesions in baseline CT.

Note: Measuring breakdown characteristics during the hot re-ignition of high intensity discharge lamps using high frequency alternating current voltage.

To investigate the cold and hot re-ignition properties of High Intensity Discharge (HID) lamps in more detail an automated setup was designed in such a way that HID lamps of various sizes and under different background pressures can be tested. The HID lamps are ignited with a ramped sinusoidal voltage signal with frequencies between 60 and 220 kHz and with amplitude up to 7.5 kV. Some initial results of voltage and current measurements on a commercially available HID lamp during hot and cold re-ignition are presented.

Quality of life in thalassemia: a comparison of SF-36 results from the thalassemia longitudinal cohort to reported literature and the US norms.

Thalassemia is a chronic, inherited blood disorder, which, in its most severe form, causes life-threatening anemia. Advances in treatment have led to increased life expectancy however the need for chronic blood transfusions and chelation therapy remains a significant burden for patients. Our study compared health related quality of life (HRQOL) from the Thalassemia Clinical Research Network's (TCRNs) Thalassemia Longitudinal Cohort (TLC) study to US norms and assessed association with clinical variables. There were 264 patients over age 14 who completed the Medical Outcomes Study 36-Item Short Form Health Survey version 2 (SF36v2) baseline assessment. When compared to US norms, TLC patients had statistically significant (P < 0.05) worse HRQOL on five of the eight subscales (physical functioning, role-physical, general health, social functioning, and role-emotional) and on both summary scales (physical component summary and mental component summary). Women, older patients, and those with more disease complications and side effects from chelation reported lower HRQOL. In general, adolescents and adults with thalassemia report worse HRQOL than the US population, despite contemporary therapy. The SF-36 should become a standard instrument for assessing HRQOL in thalassemia to determine predictors of low HRQOL which may be better addressed by a multidisciplinary team.

Self-evaluation in dialysis patients.

OBJECTIVE: The aim of this study is to present the results of a self-reported evaluation of the psychoemotional status by dialysis patients. The level of self-esteem influences the emotions felt, both positive and negative, which in turn may determine the adherence to treatment instructions, and which certainly is reflected in the somatic condition. MATERIAL AND METHODS: The study was a randomized controlled trial using a sample of 102 fully informed and consenting patients with end-stage renal disease and 102 people from the general population. The survey instrument used was a Self-Esteem Inventory. RESULTS: The results show that there were differences between the dialysis patients and the general population concerning Physical Self-Esteem and Acting/Task Self-Esteem. The patients scored significantly lower than the healthy subjects lower on both subscales. No differences were noted between the two groups of subjects regarding of Social Self-Esteem and Emotional Self-Esteem. The results also show that the level of self-esteem in dialyzed patients under the age 50 years was higher than in those above 50 regarding the sociability, sense of humor, memory, and the sense of being accepted by others. CONCLUSION: We conclude that there are differences in the self-reported level of self-esteem between dialyzed patients and the general population. The patients' age also factors in the self-reported assessment.

Sequential translocation of tyrosine kinases Lyn and Syk to the activated Fcgamma receptors during phagocytosis.

Tyrosine phosphorylation of numerous proteins is one of the earliest events detectable during Fcgamma receptor-mediated phagocytosis. We demonstrate that IgG-coated particles associated with the surface of macrophages are enriched with numerous tyrosine-phosphorylated proteins. During particle internalization the proteins are still associated with particles but their phosphorylation is reduced. Lyn kinase is phosphorylated both at particle binding and internalization steps. The phosphorylated Syk kinase is the major kinase associated with engulfed particles. Imnunofluorescent studies confirm spatial and temporal distribution of Lyn and Syk kinases at different stages of phagocytosis. Our data indicate that ligation of Fcgamma receptors activates Lyn followed by Syk kinase and in the result multimolecular complex of the kinases and several accompanying tyrosine phosphorylated proteins with Fcgamma receptors is organized leading to local reorganization of actin-based skeleton and particle uptake.

The clustered Fcgamma receptor II is recruited to Lyn-containing membrane domains and undergoes phosphorylation in a cholesterol-dependent manner.

Phosphorylation of clustered Fcgamma receptor II (FcgammaRII) by Src family tyrosine kinases is the earliest event in the receptor signaling cascade. However, the molecular mechanisms for the interaction between FcgammaRII and these kinases are not elucidated. To asses this problem we isolated high molecular weight complexes of cross-linked FcgammaRII from non-ionic detergent lysates of U937 monocytic cells. CD55, a glycosylphosphatidylinositol-anchored protein, a ganglioside GM1 and Lyn, a Src family tyrosine kinase, were also located in these complexes. Gradient centrifugation demonstrated that the complexes containing cross-linked FcgammaRII displayed a low buoyant density. The FcgammaRII present in the complexes underwent tyrosine phosphorylation. Cross-linked FcgammaRII and Lyn occupied common 100-200 nm detergent-resistant membrane fragments, as demonstrated by immunoprecipitation and microscopy studies. Pretreatment of the cells with beta-cyclodextrin, a cholesterol acceptor, depleted membrane cholesterol and released CD55, GM1 and Lyn from the detergent-resistant complexes. In parallel, the association of Lyn with cross-linked FcgammaRII was disrupted and phosphorylation of the receptor inhibited. Reincorporation of cholesterol evoked the relocation of Lyn into the detergent-resistant membrane fraction and restored both Lyn association with cross-linked FcgammaRII and tyrosine phosphorylation of the receptor. Our data demonstrate that cholesterol-enriched membrane rafts can facilitate tyrosine phosphorylation of clustered FcgammaRII by Lyn kinase.

The role of cholesterol and sphigomyelin in tyrosine phosphorylation of proteins and capping of Fcgamma receptor II.

Cross-linking of cell surface receptors by multivalent ligands, e.g. by antibodies, evokes their clustering -- patching. Subsequently, these clusters can be translocated by the acto-myosin machinery toward one pole of the cell and assembly cap. Patching of FcgammaRII in U937 cells correlates with tyrosine phosphorylation of several proteins while cap assembly correlates with their dephosphorylation. To study the mechanism of activation of tyrosine kinases during FcgammaRII activation we disturbed the organization of the putative plasma membrane microdomains by depletion of membrane cholesterol and sphingomyelin. Cholesterol was removed with the use of beta-cyclodextrin while sphingomyelin was decomposed by exogenous sphingomyelinase. Cyclodextrin at 5-10 mM removed about 70% of cholesterol from the cells and abolished the assembly of FcgammaRII caps thereby arresting the receptors at the patching stage. Similarly, 70 mU/ml sphingomyelinase inhibited cap formation by 60%. Cholesterol and sphingomyelin depletion also suppressed the tyrosine phosphorylation of proteins which accompanied cross-linking of FcgammaRII. The observations indicate that cholesterol and sphingomyelin can control the interactions of tyrosine kinases with clustered FcgammaRII.

Engagement of spectrin and actin in capping of FcgammaRII revealed by studies on permeabilized U937 cells.

Plasma membrane receptors can undergo translocation in the plane of plasma membrane after binding of polyvalent ligands. Ligand/receptor clusters, named patches, can collect into a polar cap, presumably due to their association with the submembrane actin-based cytoskeleton. We found that the assembly of Fcgamma receptor II caps in human monocytic U937 cells was accompanied by the accumulation of spectrin and actin in the cap region. Permeabilization of cells with streptolysin O rendered capping sensitive to inhibition by phalloidin, an actin filament stabilizing agent. A rabbit antibody directed against the chicken erythrocyte alpha-subunit of spectrin, an actin- and membrane-binding protein, also blocked the capping in a dose dependent manner. The inhibition reached approximately 50% after 20 minutes of cell treatment with the antibody. Anti-alpha-spectrin targeted specifically its submembrane antigen, in contrast to unspecific antibodies which remained dispersed in the cell interior and had no influence on the cap assembly. Our results indicate an active engagement of spectrin and actin filaments in the capping of Fcgamma receptor II.

Signaling pathways in phagocytosis.

Phagocytosis is an uptake of large particles governed by the actin-based cytoskeleton. Binding of particles to specific cell surface receptors is the first step of phagocytosis. In higher Eucaryota, the receptors able to mediate phagocytosis are expressed almost exclusively in macrophages, neutrophils, and monocytes, conferring immunodefence properties to these cells. Receptor clustering is thought to occur upon particle binding, that in turn generates a phagocytic signal. Several pathways of phagocytic signal transduction have been identified, including the activation of tyrosine kinases and (or) serine/threonine kinase C in pivotal roles. Kinase activation leads to phosphorylation of the receptors and other proteins, recruited at the sites of phagocytosis. Monomeric GTPases of the Rho and ARF families are likely to be engaged downstream of activated receptors. The GTPases, in cooperation with phosphatidylinositol 4-phosphate 5-kinase and phosphatidylinositol 3-kinase lipid modifying enzymes, can modulate locally the assembly of the submembranous actin filament system leading to particle internalization.

Tyrosine phosphorylation/dephosphorylation controls capping of Fcgamma receptor II in U937 cells.

In the capping of cell-surface receptors two stages can be distinguished: 1) clustering of the receptors (patching) induced by cross-linking with specific antibodies and 2) subsequent assembly of patches into a cap which is driven by the actin-based cytoskeleton. We found that patching of Fcgamma receptor II in U937 cells was correlated with tyrosine phosphorylation of certain proteins, most prominently those of 130, 110, 75 and 28 kDa. The phosphotyrosine-bearing proteins were accumulated at the receptor patches. Formation of the receptor caps was coincident with dephosphorylation of these proteins. Inhibition of protein tyrosine kinases with herbimycin A and genistein attenuated the protein tyrosine hyperphosphorylation and blocked capping in a dose-dependent manner. Phenylarsine oxide and pervanadate, inhibitors of protein tyrosine phosphatases, also suppressed capping of Fcgamma receptor II in a concentration-dependent fashion. Simultaneously, tyrosine hyperphosphorylation of proteins occurred. In the presence of the tyrosine kinase and phosphatase inhibitors the receptors were arrested at the patching stage. In contrast, okadaic acid, a serine/threonine phosphatase blocker, did not affect assembly of the receptor caps. The inhibitory effect of phenylarsine oxide was rapidly reversed by dithiols, 2,3-dimercapto-1-propanoldithiol and dithiotreitol, and was coincident with dephosphorylation of protein tyrosine residues. Extensive washing of pervanadate-exposed cells also resulted in progressive restoration of the cap assembly. Using streptolysin O-permeabilized cells we confirmed regulatory function played by dephosphorylation of tyrosine residues in capping of Fcgamma receptor II. Exogenous phosphatases, applied to permeabilized cells in which activity of endogenous tyrosine phosphatases was blocked, evoked dephosphorylation of protein tyrosine residues that was accompanied by recovery of capping ability in the cells.

Tyrosine phosphorylation and Fcgamma receptor-mediated phagocytosis.

Phagocytosis of IgG-opsonized particulate material in hematopoietic cells is mediated by Fcgamma receptors (FcgammaRs). Interaction of the receptors with Fc domains of IgG triggers transduction of phagocytic signal in which a key role is played by phosphorylation of tyrosine residues of the receptors. These residues are arranged into a specific motif (immunoreceptor tyrosine-based activation motif; ITAM) which is located either in the cytoplasmic part of FcgammaRIIA or in gamma chains associated with FcgammaRI and FcgammaRIIIA. The conserved tyrosine residues are phosphorylated by, and associate with, tyrosine kinases of Src and Syk families. Coordinated action of these components initiates numerous intracellular events leading finally to local rearrangement of the actin-based cytoskeleton and internalization of the particles.

Syk kinase, tyrosine-phosphorylated proteins and actin filaments accumulate at forming phagosomes during Fcgamma receptor-mediated phagocytosis.

Phagocytosis mediated by Fcgamma receptors (FcgammaRs) is thought to be regulated by a cascade of tyrosine phosphorylation events that finally leads to the rearrangement of submembranous actin-based cytoskeleton and internalization of particles. Suggestions concerning the functional relationship between protein tyrosine kinases, their substrates, and actin filament reorganization prompted us to determine cellular distribution of these elements during uptake of IgG-coated particles in murine thio-macrophages. We found that the onset of uptake of the particles was accompanied by tyrosine phosphorylation of several proteins, among which 90, 50, 40, 30, and 25 kDa polypeptides were distinguished. In most of the proteins the tyrosine hyperphosphorylation persisted up to 3 min of the uptake; however, kinetics of the phosphorylation of individual proteins varied. Immunofluorescence data showed that the phosphotyrosine-bearing proteins were localized in regions of the particle uptake, being concentrated at phagocytic cups and nascent phagosomes. The local enrichment in tyrosine phosphorylated proteins was correlated with accumulation of actin filaments at these early stages of phagosome formation. During phagosome maturation, both tyrosine phosphorylated proteins and microfilaments disappeared from the periphagosomal regions. Syk, one of the tyrosine kinases, was translocated to the regions where FcgammaR-mediated phagocytosis had started. On the contrary, no enrichment in phosphatidylinositol 3-kinase was detected in these places.

Local accumulation of alpha-spectrin-related protein under plasma membrane during capping and phagocytosis in Acanthamoeba.

During capping and phagocytosis the interaction between cluster cell surface receptors and the submembraneous actin-based skeleton may be mediated by spectrin-like proteins. To test this possibility we examined the localization of an alpha-spectrin immunoanalogue, that had been previously identified in whole extracts of Acanthamoeba, during capping of Con A receptors and during phagocytosis of Con A-coated yeast. During capping alpha-spectrin and filamentous actin co-migrated with the Con A receptors and accumulated in the region of cap formation, as demonstrated by double immunofluorescence studies. Immunoelectron microscopy revealed submembraneous location of alpha-spectrin in cells exposed to Con A, both at the time of initial cross-linking and during accumulation of alpha-spectrin in the region of the cap. Phagocytosis studies showed that alpha-spectrin and actin filaments were concentrated around phagocytic cups that enclosed ConA-coated yeast upon internalization. The proteins also surrounded nascent phagosomes present in the vicinity of the plasma membrane but were absent at the later time point of phagosome maturation. These data demonstrate a correlation between clustering of cell surface receptors and submembraneous localization of alpha-spectrin, suggesting an involvement of spectrin-like proteins in mediating the interaction of receptor clusters with the actin cytoskeleton.

Interaction of annexins IV and VI with phosphatidylserine in the presence of Ca2+: monolayer and proteolytic study.

Annexins, Ca2+- and phospholipid-binding proteins are known to bind to artificial and biological membranes in a calcium-dependent manner. However, the precise mechanism of the annexin-membrane interactions still remains to be studied in detail. In this paper we describe the results of studies on the interactions of the annexin/Ca complexes with phospholipids, obtained by the Wilhelmy balance method of assessing the surface pressure of a phospholipid monolayer. We show that the annexin IV/Ca as well as annexin VI/Ca complexes significantly reduce the surface pressure of a phosphatidylserine monolayer, when its initial value is close to collapse pressure. The effect is highly specific for monolayers composed of phosphatidylserine and strongly sensitive to pH and ionic strength. The most pronounced changes have been observed at pH 7.0-7.5, at a protein/Ca molar ratio of 1:2 for annexin IV and 1:4 for annexin VI. In the presence of sodium chloride at concentrations exceeding 400mM this effect was almost completely abolished. The obtained results point to the mainly electrostatic character of the annexin/phosphatidylserine interactions. In addition, using large multilamellar lipid vesicles and serine proteases, we demonstrate that annexins, when bound in a ternary complex with phospholipids and calcium ions, are partially protected against proteolysis. Our observation that annexin molecules, complexed with calcium ions, are protected against proteolytic attack in the presence of PS liposomes does not have to be necessarily explained in terms of partial penetration of protein within the membrane bilayer.

Phagocytosis occurs in Acanthamoeba castellanii after electroporation.

Acanthamoeba cells treated with an electric discharge were porated and their cytoplasm became accessible to exogenous molecules. Over a broad range of electric field densities low molecular weight markers (trypan blue, ruthenium red), normally unable to penetrate a plasma membrane, gained access to cytoplasm of 80-90% of the cells. Macromolecules (albumin-FITC and IgG-FITC) penetrated into 63-86% of the cells when electroporation was carried out over the range of 1500V/25 microF-400V/250 microF. Pulse labeling with fluorescent markers evidenced that even 3 hrs. after an electric pulse the plasma membrane was still permeable to exogenous fluorescent probes. Following this stage, the pores were gradually closed. The cells electroporated at 400 V/250 microF were able to ingest yeast particles. The uptake of the particles seems to be an active process since it was inhibited by azide and phalloidin. Therefore, the electroporation of Acanthamoeba makes possible the introduction of macromolecules into the cells and subsequent analysis of their effect on active motile processes such as phagocytosis. This should greatly facilitate characterization of the mechanisms by which such processes do occur.