Production and Characterization of Peptide Antibodies to the C-Terminal of Frameshifted Calreticulin Associated with Myeloproliferative Diseases

Myeloproliferative Neoplasms (MPNs) constitute a group of rare blood cancers that are characterized by mutations in bone marrow stem cells leading to the overproduction of erythrocytes, leukocytes, and thrombocytes. Mutations in calreticulin (CRT) genes may initiate MPNs, causing a novel variable polybasic stretch terminating in a common C-terminal sequence in the frameshifted CRT (CRTfs) proteins. Peptide antibodies to the mutated C-terminal are important reagents for research in the molecular mechanisms of MPNs and for the development of new diagnostic assays and therapies. In this study, eight peptide antibodies targeting the C-terminal of CRTfs were produced and characterised by modified enzyme-linked immunosorbent assays using resin-bound peptides. The antibodies reacted to two epitopes: CREACLQGWTE for SSI-HYB 385-01, 385-02, 385-03, 385-04, 385-07, 385-08, and 385-09 and CLQGWT for SSI-HYB 385-06. For the majority of antibodies, the residues Cys¹, Trp⁹, and Glu¹¹ were essential for reactivity. SSI-HYB 385-06, with the highest affinity, recognised recombinant CRTfs produced in yeast and the MARIMO cell line expressing CRTfs when examined in Western immunoblotting. Moreover, SSI-HYB 385-06 occasionally reacted to CRTfs from MPN patients when analysed by flow cytometry. The characterized antibodies may be used to understand the role of CRTfs in the pathogenesis of MPNs and to design and develop new diagnostic assays and therapeutic targets.     

A rapid interview protocol supporting patient-centered quality improvement: hearing the parent's voice in a pediatric cancer unit

BACKGROUND: The Institute of Medicine's 2001 report on quality delimits six dimensions of optimal care: safety, effectiveness, efficiency, timeliness, patient centeredness, and equity. In fall 2001 parents of pediatric cancer patients were interviewed to determine how well they thought these dimensions were addressed with respect to medication administration. Immediate goals were to identify system weaknesses and devise strategies to prevent future errors. A higher-order goal was to develop and demonstrate a model protocol for rapid-cycle interview assessments. METHODS: Hematology/oncology directors worked with a research expert to develop a semistructured interview protocol. After training, which included directed reading, oral instruction, and role-playing, a convenience sample of 20 English- and Spanish-speaking parents of inpatients was recruited. Parents were asked to characterize current medication administration practices and to describe problems that they had experienced or witnessed. Rapid content analysis techniques were used to identify issues of importance to the parents. FINDINGS: Parents' medication concerns centered on their children's comfort. Parents called for communication improvements, standardization of all nursing procedures and techniques, and a guide or an outline providing a clear understanding of what to expect when and from whom. Viewing these concerns in relation to the Institute of Medicine's quality domains allowed the department to frame an improvement action plan aligned with organizational and national priorities. IMPLICATIONS: With good supervision and limited focused training, inexperienced staff can successfully administer semistructured qualitative interviews and help analyze findings for rapid cycle improvement purposes. The protocol can be adapted for use in organizations interested in rapid qualitative assessments of patient and parent preferences.     

Surface topography and tribological properties of coatings prepared from microparticle size polyurethane dispersions studied by atomic force microscopy

The surface topography and mechanical properties of coatings prepared using large particle size polyurethane dispersions (PUD) are investigated using atomic force microscopy (AFM) imaging, AFM-based force measurements, and friction force microscopy. PUD coatings, which are prepared from dispersions containing particles of micron size, have surface roughness of 250–300 nm and waviness of 2.5–3 μm resulting from the particle size. The surface moduli of the PUD coatings are varied by tuning the ratio of hard-to-soft segmentation in the polyurethanes and are found to be between 40 and 100 MPa. The friction coefficient obtained in the study is found to be correlated with both the surface modulus of the coatings and the adhesion between the probe and the samples and is well in line with the perceived feel of an experienced human panel. The data are very well behaved and clearly show the utility of this technique in characterizing these types of surfaces.     

Catalytic wall reactor: Catalytic coatings of stainless steel by VOx/TiO2 and Co/SiO2 catalysts

Catalytic wall (structured) reactors and structured supports are suitable to study the catalytic properties of nanosized materials. The coating of metallic (aluminum and stainless steel) plates by thin layers of active phase is presented in two cases, VO_x/TiO₂ and Co/SiO₂, catalysts used in the oxidative dehydrogenation (ODH) of propane and in Fischer–Tropsch synthesis (FTS) of clean fuels, respectively. The preparation of coated plates and their characterisation by various methods of physicochemical analysis are described. Both chemical and physical methods were used for coating. VO_x/TiO₂ layers were obtained by grafting of Ti (on Al or stainless-steel plates) and V (on TiO₂) alkoxides and use of sol–gel media or suspension. A silica primer was deposited (on stainless-steel plate) by plasma-assisted chemical vapour deposition (PACVD) onto which Co oxide and silica were coprecipitated from sol–gel. The catalytic experiments in the respective reactions were carried out in special plate reactors and compared with those of catalytic powders. The study shows that the coating of a metallic substrate by a catalyst is not straightforward and requires specific studies dealing with both chemistry (chemical affinity between substrate and catalytic layers) and catalytic engineering (catalytic performance in taylor-made reactors).     

31P MAS NMR Spectroscopy of Hexachlorocyclotriphosphazene at Different Stages During Thermal Ring-Opening Polymerization

The basic concepts of solid-state nuclear magnetic resonance (NMR) spectroscopy are explained in a clear and concise manner. First the reader is introduced to the principles of NMR spectroscopy and techniques that are most commonly utilized today. Next, an overview of advanced magic-angle spinning (MAS) NMR experiments, which can provide valuable insight into molecular structure, connectivity, dynamics, domains, and phases of inorganic polymers, is presented. The final chapter discusses the major classes of inorganic polymers, their properties and applications. It addresses their characterization including, if any, the role of solution and solid-state NMR spectroscopy, and provides prospective on potential applications of these techniques for the given class of material.     

Kinetics of emitted mass—A study with three dry powder inhaler devices

The purpose of this study was to understand the kinetics of emptying of micronised salbutamol sulphate (SS) and lactohale 300 (LH300) under varying air flow rates (30–180 L min⁻¹) from three dry powder inhaler devices, Rotahaler^® (RH), Monodose Inhaler^® (MI) and Handihaler^® (HH). Aerosol concentration vs. time profiles, determined through real-time laser diffraction particle sizing, demonstrated that the majority of the powder from RH was emitted within one second while a more prolonged aerosolisation was observed from MI and HH. Peak aerosol concentrations were achieved more rapidly from RH compared to MI for both SS and LH300. Calculated cumulative emitted mass (CCEM) vs. time profiles were obtained from the aerosol concentration vs. time profiles and the emitted mass and the rate of CCEM (k_CCEM), estimated from a modelling approach, increased with increasing air flow rates. The k_CCEMvs. air flow rate profiles of SS and LH300 were significantly different at high air flow rates. The k_CCEM was highest from RH and lowest from MI. Differences in k_CCEM between the devices were related to capsule aperture size while the differences between the materials were due to the powder bed structure. This approach provided an understanding of the rate at which powder mass emptied from dry powder inhaler devices and was proposed to be a powerful development tool for the future powder inhalers.