Evaluation of ELISA procedures to detect von Willebrand Factor with monoclonal antibodies

The von Willebrand Factor is a multimeric glycoprotein responsible for the promotion of platelet adhesion and aggregation at sites of vascular injury, and for FVIII stabilization. Abnormalities on this protein are responsible for diverse types of von Willebrand Disease. In the present study, monoclonal antibodies against human von Willebrand Factor were developed as a means to improve von Willebrand Disease research and diagnosis. Monoclonal antibodies were tested for their ability to bind to purified and plasmatic von Willebrand Factor. Monoclonal antibodies vW22 and vW23 were found to bind only purified von Willebrand Factor, and monoclonal antibodies vW18 and vW21 were found to bind purified and plasmatic von Willebrand Factor. Antibodies vW18 and vW21 were used to perform a sandwich-enzyme-linked immunosorbent assay to detect and quantify von Willebrand Factor concentration in plasma samples from 143 coagulopathy patients and 12 healthy blood donors. The assay showed high performance, with strong correlation and agreement in results, when compared to electroimmunoassay (Rs = 0.843 and K = 0.691 with p<0.001) and a commercial ELISA (Rs = 0.930 and K = 0.819 with p<0.001). S-ELISA proved to be a useful tool in vWF quantification tests in Brazilian specialized laboratories as an alternative to imported tests.(Avaliação de procedimentos ELISA para detectar o Fator von Willebrand com anticorpos monoclonais). O Fator von Willebrand é uma glicoproteína multimérica responsável pela promoção da adesão e agregação de plaquetas nos locais de lesão vascular e pela estabilização do FVIII. Anormalidades na função ou estrutura desta proteína são responsáveis por diversos tipos de doença de von Willebrand. Para auxiliar na pesquisa e diagnóstico da doença de von Willebrand, este estudo desenvolveu anticorpos monoclonais contra fator von Willebrand humano. Os anticorpos monoclonais foram testados quanto à sua capacidade de se ligar ao fator von Willebrand purificado e plasmático. Os anticorpos monoclonais vW22 e vW23 ligaram-se somente ao fator von Willebrand purificado, e anticorpos monoclonais vW18 e vW21 ligaram-se tanto ao fator von Willebrand purificado como ao plasmático. Anticorpos vW18 e vW21 foram utilizados no desenvolvimento de um ELISA sandwich para detectar e quantificar a concentração de fator von Willebrand no plasma em uma amostra de 143 pacientes com coagulopatias e 12 doadores de sangue saudáveis. O teste mostrou alto desempenho, com forte correlação e concordância quando comparado com uma imunoeletrofose (Rs = 0,843 e K = 0,691 p <0,001) e um ELISA comercial (Rs = 0,930 e K = 0,819 p <0,001). O ELISA-S desenvolvido no presente trabalho mostrou um bom desempenho para ser usado como teste de quantificação vWF em laboratórios especializados no Brasil como alternativa para testes importados

Технология индивидуализации обучения иностранным языкам в средней школе

Цель работы: разработать приемы индивидуализации при обучении иностранному языку в средней школе и апробировать ее опытно-экспериментальным путем. Объектом исследования является процесс обучения иностранному языку в средней школе. Предмет исследования: приемы индивидуализации обучения иностранному языку в средней школе

Temporal and effort cost decision-making in healthy individuals with subclinical psychotic symptoms

The value people attribute to rewards is influenced both by the time and the effort required to obtain them. Impairments in these computations are described in patients with schizophrenia and appear associated with negative symptom severity. This study investigated whether deficits in temporal and effort cost computations can be observed in individuals with subclinical psychotic symptoms (PS) to determine if this dysfunction is already present in a potentially pre-psychotic period. Sixty participants, divided into three groups based on the severity of PS (high, medium and low), performed two temporal discounting tasks with food and money and a concurrent schedule task, in which the effort to obtain food increased over time. We observed that in high PS participants the discounting rate appeared linear and flatter than that exhibited by participants with medium and low PS, especially with food. In the concurrent task, compared to those with low PS, participants with high PS exerted tendentially less effort to obtain snacks only when the required effort was high. Participants exerting less effort in the higher effort condition were those with higher negative symptoms. These results suggest that aberrant temporal and effort cost computations might be present in individuals with subclinical PS and therefore could represent a vulnerability marker for psychosis

Size and Composition Distribution of Atmospheric Particles in Southern California

Continuous measurements of single particle size and chemical composition in the atmosphere are made using aerosol time-of-flight mass spectrometers (ATOFMS) operated alongside more conventional reference air sampling instruments at a network of three urban air monitoring sites in southern California. Electrical aerosol analyzers and optical particle counters are employed to acquire continuous particle size distribution data, and inertial impactor and bulk filter samples with 4-h resolution are taken for determination of particle size and chemical composition. Filter and impactor samples also are taken upwind of the air basin at Santa Catalina Island in order to characterize background air quality. The airborne particle size and composition distribution as measured by the cascade impactors at inland sites differ from that over the ocean principally due to depletion of sea salt particles accompanied by the addition of fine carbon-containing particles and secondary aerosol nitrate. Data from the ATOFMS systems create a continuous time series of sodium-, ammonium-, nitrate-, and carbon-containing particle counts that provide a high-resolution view of differences in particle composition as a function of location in the air basin. Results show that the characteristic peak in the Los Angeles area aerosol mass distribution in the 0.2−0.3-μm size range observed during the 1987 SCAQS experiments has been reduced, consistent with reductions in diesel soot and elemental carbon emissions since that time

Size and Composition Distribution of Atmospheric Particles in Southern California

Continuous measurements of single particle size and chemical composition in the atmosphere are made using aerosol time-of-flight mass spectrometers (ATOFMS) operated alongside more conventional reference air sampling instruments at a network of three urban air monitoring sites in southern California. Electrical aerosol analyzers and optical particle counters are employed to acquire continuous particle size distribution data, and inertial impactor and bulk filter samples with 4-h resolution are taken for determination of particle size and chemical composition. Filter and impactor samples also are taken upwind of the air basin at Santa Catalina Island in order to characterize background air quality. The airborne particle size and composition distribution as measured by the cascade impactors at inland sites differ from that over the ocean principally due to depletion of sea salt particles accompanied by the addition of fine carbon-containing particles and secondary aerosol nitrate. Data from the ATOFMS systems create a continuous time series of sodium-, ammonium-, nitrate-, and carbon-containing particle counts that provide a high-resolution view of differences in particle composition as a function of location in the air basin. Results show that the characteristic peak in the Los Angeles area aerosol mass distribution in the 0.2−0.3-μm size range observed during the 1987 SCAQS experiments has been reduced, consistent with reductions in diesel soot and elemental carbon emissions since that time

Cellular magnesium acquisition : an anomaly in embryonic cation homeostasis

Author Posting. © The Author(s), 2007. This is the author's version of the work. It is posted here by permission of Elsevier B.V. for personal use, not for redistribution. The definitive version was published in Experimental and Molecular Pathology 83 (2007): 224-240, doi:10.1016/j.yexmp.2007.03.007.The intracellular dominance of magnesium ion makes clinical assessment difficult despite the critical role of Mg++ in many key functions of cells and enzymes. There is general consensus that serum Mg++ levels are not representative of the growing number of conditions for which magnesium is known to be important. There is no consensus method or sample source for testing for clinical purposes. High intracellular Mg++ in vertebrate embryos results in part from interactions of cations which influence cell membrane transport systems. These are functionally competent from the earliest stages, at least transiently held over from the unfertilized ovum. Kinetic studies with radiotracer cations, osmolar variations, media lacking one or more of the four biological cations, Na+, Mg++, K+, and Ca++, and metabolic poison 0.05 mEq/L NaF, demonstrated: (1) all four cations influence the behavior of the others, and (2) energy is required for uptake and efflux on different time scales, some against gradient. Na+ uptake is energy dependent against an efflux gradient. The rate of K+ loss is equal with or without fluoride, suggesting a lack of an energy requirement at these stages. Ca++ efflux took twice as long in the presence of fluoride, likely due in part to intracellular binding. Mg++ is anomalous in that early teleost vertebrate embryos have an intracellular content exceeding the surrounding sea water, an isolated unaffected yolk compartment, and a clear requirement for energy for both uptake and efflux. The physiological, pathological, and therapeutic roles of magnesium are poorly understood. This will change: (1) when 28Mg is once again generally available at a reasonable cost for both basic research and clinical assessment, and (2) when serum or plasma levels are determined simultaneously with intracellular values, preferably as part of complete four cation profiles. Atomic absorption spectrophotometry, energy-dispersive x-ray analysis, and inductively coupled plasma emission spectroscopy on sublingual mucosal and peripheral blood samples are potential methods of value for coordinated assessments.AEC Grant No. 134

The Youngest Victims: Children and Youth Affected by War

In 1989, the United Nation Convention on the Rights of the Child declared, “[state parties] shall take all feasible measures to ensure protection and care of children who are affected by an armed conflict.” In addition to attempting to secure the welfare of children in armed conflict, the Convention went on to ban the recruitment and deployment of children during armed conflict. Despite the vast majority of sovereign nations signing and ratifying this agreement, this treaty, unfortunately, has not prevented children and youth from witnessing, becoming victims of, or participating in political, ethnic, religious, and cultural violence across the past three decades. This chapter offers an “ecological perspective” on the psychosocial consequences of exposure to the trauma of war-related violence and social disruption

AD51B in Familial Breast Cancer

Common variation on 14q24.1, close to RAD51B, has been associated with breast cancer: rs999737 and rs2588809 with the risk of female breast cancer and rs1314913 with the risk of male breast cancer. The aim of this study was to investigate the role of RAD51B variants in breast cancer predisposition, particularly in the context of familial breast cancer in Finland. We sequenced the coding region of RAD51B in 168 Finnish breast cancer patients from the Helsinki region for identification of possible recurrent founder mutations. In addition, we studied the known rs999737, rs2588809, and rs1314913 SNPs and RAD51B haplotypes in 44,791 breast cancer cases and 43,583 controls from 40 studies participating in the Breast Cancer Association Consortium (BCAC) that were genotyped on a custom chip (iCOGS). We identified one putatively pathogenic missense mutation c.541C>T among the Finnish cancer patients and subsequently genotyped the mutation in additional breast cancer cases (n = 5259) and population controls (n = 3586) from Finland and Belarus. No significant association with breast cancer risk was seen in the meta-analysis of the Finnish datasets or in the large BCAC dataset. The association with previously identified risk variants rs999737, rs2588809, and rs1314913 was replicated among all breast cancer cases and also among familial cases in the BCAC dataset. The most significant association was observed for the haplotype carrying the risk-alleles of all the three SNPs both among all cases (odds ratio (OR): 1.15, 95% confidence interval (CI): 1.11–1.19, P = 8.88 x 10−16) and among familial cases (OR: 1.24, 95% CI: 1.16–1.32, P = 6.19 x 10−11), compared to the haplotype with the respective protective alleles. Our results suggest that loss-of-function mutations in RAD51B are rare, but common variation at the RAD51B region is significantly associated with familial breast cancer risk