Laboratory analyses for evaluation of platelet disorders and platelet concentrates

High quality of platelet analytics requires specialized knowledge and skills. It was applied to analyze platelet activation and aggregation responses in a prospective controlled study of patients with Finnish type of amyloidosis. The 20 patients with AGel amyloidosis displayed a delayed and more profound platelet shape change than healthy siblings and healthy volunteers, which may be related to altered fragmentation of mutated gelsolin during platelet activation. Alterations in platelet shape change have not been reported in association with platelet disorders. In the rare Bernard-Soulier syndrome with Asn45Ser mutation of glycoprotein (GP) IX, the diagnostic defect in the expression of GPIb-IX-V complex was characterized in seven Finnish patients, also an internationally exceptionally large patient series. When measuring thrombopoietin in serial samples of amniotic fluid and cord blood of 15 pregnant women with confirmed or suspected fetal alloimmune thrombocytopenia, the lower limit of detection could be extended. The results approved that thrombopoietin is present already in amniotic fluid. The application of various non-invasive means for diagnosing thrombocytopenia (TP) revealed that techniques for estimating the proportion of young, i.e. large platelets, such as direct measurement of reticulated platelets and the mean platelet size, would be useful for evaluating platelet kinetics in a given patient. Due to different kinetics between thrombopoietin and increase of young platelets in circulation, these measurements may have most predictive value when measured from simultaneous samples. Platelet autoantibodies were present not only in isolated autoimmune TP but also in patients without TP where disappearance of platelets might be compensated by increased production. The autoantibodies may also persist after TP has been cured. Simultaneous demonstration of increased young platelets (or increased mean platelet volume) in peripheral blood and the presence of platelet associated IgG specificities to major glycoproteins (GPIb-IX and GPIIb-IIIa) may be considered diagnostic for autoimmune TP. Measurement of a soluble marker as a sign of thrombin activation and proceeding deterioration of platelet components was applied to analyze the alterations under several stress factors (storage, transportation and lack of continuous shaking under controlled conditions) of platelet products. The GPV measured as a soluble factor in platelet storage medium showed good correlation with an array of other measurements commonly applied in characterization of stored platelets. The benefits of measuring soluble analyte in a quantitative assay were evident.

Experimental studies on the wear of engine components

Wear studies of engine components of high-speed diesel engines running under various operating conditions are presented. Tests were conducted under controlled conditions over long periods. The results of the various tests are discussed and attempts have been made to examine the effects of engine operating variables and the quality of the lubricating oil on the wear of engine components.

Significant effects of Pgi genotype and body reserves on lifespan in the Glanville fritillary butterfly

Individuals with a particular variant of the gene phosphoglucose isomerase (Pgi ) have been shown to have superior dispersal capacity and fecundity in the Glanville fritillary butterfly (Melitaea cinxia), raising questions about the mechanisms that maintain polymorphism in this gene in the field. Here, we investigate how variation in the Pgi genotype affects female and male life history under controlled conditions. The most striking effect is the longer lifespan of genotypes with high dispersal capacity, especially in nonreproducing females. Butterflies use body reserves for somatic maintenance and reproduction, but
different resources (in thorax versus abdomen) are used under dissimilar conditions, with some interactions with the Pgi genotype. These results indicate life-history trade-offs that involve resource allocation and genotype!environment interactions, and these trade-offs are likely to contribute to the maintenance of Pgi polymorphism in the natural populations.

The interaction of glycoxylate with cysteine and its application to the assay of isocitritase and of transaminases involving glyoxylate

An interesting interaction between glyoxylate and cystein takes place in phosphate buffer (pH 7.0) to form a product which is resistant to hydrolysis at ordinary temperatures. The reaction product is broken up by acid hydrolysis at elevated temperatures under controlled conditions, giving a quantitive yield of glyoxylate. Other keto acids, such as α-ketoglutarate, pyruvate and oxaloacetate, do not interact with cysteine under similar conditions. Methods based on these findings are described for(a) direct estimation of other keto acids in the presence of glyoxylate, and (b) assay of isocitritase and glyoxylate transaminase.

Modeling of the sea surface for daylight imagery studies

Plastic-coated paper is shown to possess reflectivity characteristics quite similar to those of the surface of water. This correspondence has been used with a conversion factor to model a sea surface by means of plastic-coated paper. Such a paper model is then suitably illuminated and photographed, yielding physically simulated daylight imagery of the sea surface under controlled conditions. A simple example of sinusoidal surface simulation is described.

Enhancement of fatigue life of Ni-Ti-Fe shape memory alloys by thermal cycling

The effect of thermal cycling on the load-controlled tension-tension fatigue behavior of a Ni-Ti-Fe shape memory alloy (SMA) at room temperature was studied. Considerable strain accumulation was observed to occur in this alloy under both quasi-static and cyclic loading conditions. Though, in all cases, steady-state is reached within the first 50-100 cycles, the accumulated steady-state strain, epsilon(p.ss), is much smaller in thermally cycled alloy. As a result, the fatigue performance of them was found to be significantly enhanced vis-a-vis the as-solutionized alloy. Furthermore, under load-controlled conditions, the fatigue life of Ni-Ti-Fe alloys was found to be exclusively dependent on epsilon(p.ss). Observations made by profilometry and differential scanning calorimetry (DSC) indicate that the 200-500% enhancement in fatigue life of thermally cycled alloy is due to the homogeneous distribution of the accumulated fatigue strain. (C) 2010 Elsevier B.V. All rights reserved.

Immunochemical characterization of rapid and slowly released allergens from the pollen of Parthenium hysterophorus

Allergens from the pollen of Parthenium hysterophorus (American feverfew), responsible for high incidence of allergic rhinitis, were found by immunoprint analysis to be localized on the surface of the pollen grains. The allergens were released very rapidly when extracted in vitro. The allergenic activity of the rapid (10 s) and slowly (20 h) released pollen proteins was comparable by in vivo skin test and ELISA inhibition assay. The isoelectric focusing patterns of the rapid and slowly released proteins, were also identical. SDS-PAGE and Western blot analysis revealed that all the major pollen allergens with molecular weight 14, 28, 31, 37 and 45 kDa were eluted within 10 s of extraction. Periodate-Schiff staining showed that the 28, 31 and 45 kDa components of the pollen extract are glycoproteins. The pollen allergens released after different periods of extraction lost 75% of IgE binding activity when subjected to in situ sodium m-periodate oxidation under controlled conditions, while 80% of the allergenic activity was still retained after extensive proteolysis. Our results support the clinical observation of a rapid onset of symptoms of allergic rhinitis in patients sensitive to Parthenium pollen, mediated predominantly by glycoproteins.

Silver nanoparticles modified nanocapsules for ultrasonically activated drug delivery

Novel ultrasound-sensitive nanocapsules were designed via layer-by-layer assembly (LbL) of polyelectrolytes for remote activated release of biomolecules/drug. Nanocapsules embedded with silver nanoparticles in the walls were synthesized by alternate assembly of poly(allylamine hydrochloride) (PAH) and dextran sulfate (DS) on silica template followed by nanoparticle synthesis and subsequent template removal thus yielding nanocapsules. The silver NPs were synthesized in situ within the capsule walls under controlled conditions. The nanocapsules were found to be well dispersed and the silver NPs were evenly distributed within the shell. FITC-dextran permeated easily into the capsules containing silver NP's due to the pores generated during the formation of NP's. When the loaded nanocapsules were sonicated, the presence of the silver NPs in the shell structure led to rupturing of the shell into smaller fragments thus releasing the FITC-dextran. Such nanocapsules have the potential to be used as drug delivery vehicles and offer the scope for further development in the areas of modern medicine, material science, and biochemistry. (C) 2012 Elsevier B.V. All rights reserved.

Investigations of secondary organic aerosol formation in laboratory chambers

Secondary organic aerosol (SOA) is produced in the atmosphere by oxidation of volatile organic compounds. Laboratory chambers are used understand the formation mechanisms and evolution of SOA formed under controlled conditions. This thesis presents studies of SOA formed from anthropogenic and biogenic precursors and discusses the effects of chamber walls on suspended vapors and particles.

During a chamber experiment, suspended vapors and particles can interact with the chamber walls. Particle wall loss is relatively well-understood, but vapor wall losses have received little study. Vapor wall loss of 2,3-epoxy-1,4-butanediol (BEPOX) and glyoxal was identified, quantified, and found to depend on chamber age and relative humidity.

Particles reside in the atmosphere for a week or more and can evolve chemically during that time period, a process termed aging. Simulating aging in laboratory chambers has proven to be challenging. A protocol was developed to extend the duration of a chamber experiment to 36 h of oxidation and was used to evaluate aging of SOA produced from m-xylene. Total SOA mass concentration increased and then decreased with increasing photooxidation suggesting a transition from functionalization to fragmentation chemistry driven by photochemical processes. SOA oxidation, measured as the bulk particle elemental oxygen-to-carbon ratio and fraction of organic mass at m/z 44, increased continuously starting after 5 h of photooxidation.

The physical state and chemical composition of an organic aerosol affect the mixing of aerosol components and its interactions with condensing species. A laboratory chamber protocol was developed to evaluate the mixing of SOA produced sequentially from two different sources by heating the chamber to induce particle evaporation. Using this protocol, SOA produced from toluene was found to be less volatile than that produced from a-pinene. When the two types of SOA were formed sequentially, the evaporation behavior most closely represented that of SOA from the second parent hydrocarbon, suggesting that the structure of the mixed SOA particles resembles a core of SOA from the first precursor coated by a layer of SOA from the second precursor, indicative of limiting mixing.

Secondary Organic Aerosol Composition Studies Using Mass Spectrometry

Trace volatile organic compounds emitted by biogenic and anthropogenic sources into the atmosphere can undergo extensive photooxidation to form species with lower volatility. By equilibrium partitioning or reactive uptake, these compounds can nucleate into new aerosol particles or deposit onto already-existing particles to form secondary organic aerosol (SOA). SOA and other atmospheric particulate matter have measurable effects on global climate and public health, making understanding SOA formation a needed field of scientific inquiry. SOA formation can be done in a laboratory setting, using an environmental chamber; under these controlled conditions it is possible to generate SOA from a single parent compound and study the chemical composition of the gas and particle phases. By studying the SOA composition, it is possible to gain understanding of the chemical reactions that occur in the gas phase and particle phase, and identify potential heterogeneous processes that occur at the surface of SOA particles. In this thesis, mass spectrometric methods are used to identify qualitatively and qualitatively the chemical components of SOA derived from the photooxidation of important anthropogenic volatile organic compounds that are associated with gasoline and diesel fuels and industrial activity (C12 alkanes, toluene, and o-, m-, and p-cresols). The conditions under which SOA was generated in each system were varied to explore the effect of NO_x and inorganic seed composition on SOA chemical composition. The structure of the parent alkane was varied to investigate the effect on the functionalization and fragmentation of the resulting oxidation products. Relative humidity was varied in the alkane system as well to measure the effect of increased particle-phase water on condensed-phase reactions. In all systems, oligomeric species, resulting potentially from particle-phase and heterogeneous processes, were identified. Imines produced by reactions between (NH₄)₂SO₄ seed and carbonyl compounds were identified in all systems. Multigenerational photochemistry producing low- and extremely low-volatility organic compounds (LVOC and ELVOC) was reflected strongly in the particle-phase composition as well.

Aspects of the washout of salmonid eggs. 1. Observations on the physical characteristics of real and artificial eggs, their rate of fall in a water column and their pattern of settlement in an experimental channel

When salmonid redds are disrupted by spates, the displaced eggs will drift downstream. The mean distance of travel, the types of locations in which the eggs resettle and the depth of reburial of displaced eggs are not known. Investigation of these topics under field conditions presents considerable practical problems, though the use of artificial eggs might help to overcome some of them. Attempts to assess the similarities and/or differences in performance between real and artificial eggs are essential before artificial eggs can validly be used to simulate real eggs. The present report first compares the two types of egg in terms of their measurable physical characteristics (e.g. dimensions and density). The rate at which eggs fall in still water will relate to the rate at which they are likely to resettle in flowing water in the field. As the rate of fall will be influenced by a number of additional factors (e.g. shape and surface texture) which are not easily measured directly, the rates of fall of the two types of egg have been compared directly under controlled conditions. Finally, comparisons of the pattern of settlement of the two types of egg in flowing water in an experimental channel have been made. Although the work was primarily aimed at testing the value of artificial eggs as a simulation of real eggs, several side issues more directly concerned with the properties of real eggs and the likely distance of drift in natural streams have also been explored. This is the first of three reports made on this topic by the author in 1984.

Escapement of the Cape rock lobster (Jasus lalandii ) through the mesh and entrance of commercial traps

Metal-framed traps covered with polyethylene mesh used in the fishery for the South African Cape rock lobster (Jasus lalandii) incidentally capture large numbers of undersize (<75 mm CL) specimens. Air-exposure, handling, and release procedures affect captured rock lobsters and reduce the productivity of the stock, which is heavily fished. Optimally, traps should retain legalsize rock lobsters and allow sublegal animals to escape before traps are hauled. Escapement, based on lobster morphometric measurements, through meshes of 62 mm, 75 mm, and 100 mm was investigated theoretically under controlled conditions in an aquarium, and during field trials. SELECT models were used to model escapement, wherever appropriate. Size-selectivity curves based on the logistic model fitted the aquarium and field data better than asymmetrical Richards curves. The lobster length at 50% retention (L50) on the escapement curve for 100-mm mesh in the aquarium (75.5 mm CL) approximated the minimum legal size (75 mm CL); however estimates of L50 increased to 77.4 mm in field trials where trapentrances were sealed, and to 82.2 mm where trap-entrances were open. Therfore, rock lobsters that cannot escape through the mesh of sealed field traps do so through the trap entrance of open traps. By contrast, the wider selection range and lower L25 of field, compared to aquarium, trials (SR = 8.2 mm vs. 2.6 mm; L25 =73.4 mm vs. 74.1 mm), indicate that small lobsters that should be able to escape from 100-mm mesh traps do not always do so. Escapement from 62-mm mesh traps with open entrance funnels increased by 40−60% over sealed traps. The findings of this study with a known size distribution, are related to those of a recent indirect (comparative) study for the same species, and implications for trap surveys, commercial catch rates, and ghost fishing are discussed.

Utilization of frog waste

Commercial frog waste samples have been converted into meals by cooking at 0.7 kg/sq. cm for 30 min, draining off the stick water and drying the press cake either in the sun, tunnel dryer under controlled conditions or hot air oven. Yield of the meal varied between 18.6 to 21.5% of the fresh frog waste. Chemical analyses of the meals have shown that the meals prepared from frog waste conform to standards prescribed for fish meal and livestock feed and can therefore be used for supplementation of poultry/animal feed.

Fish tagging experiments: a prelude to an extensive tag-recovery programme on Lake Victoria

The present study is an early stage of this programme and examines several species of fishes under controlled conditions to delineate responses to tagging as a function of type of tag, species, size, and sex of fish, and position of tag placement. It is intimately related to another phase of research currently being conducted by the author on age and growth of several species important to the fishery of Lake Victoria (e.g. Tilapia spp., and several catfishes, Clarias mossambicus Peters and Bagrus docmac (Forskal). Data reported are both a reflection of growth studies and an attempt to achieve insight into tag loss, growth, and mortality that might be expected to occur in these species under lake conditions with reference to the above parameters.

Preliminary observations on cage culture of Tilapia esculenta Graham and Tilapia xillii (Gervais) in Lake Victoria waters, at the Freshwater Fisheries Institute, Nyegezi, Tanzania

Cage culture of Tilapia is not suggested as a substitute for any known techniques in fish culture, but as one of the various techniques of obtaining more fish under controlled conditions. This fact has been very well accepted in various countries. Whererever facilities exist, this line of fish culture should be vigorously explored as a possible avenue in increasing fish production. High density stocking, management under controlled conditions, easy technique of fabricating the cage at relatively low cost, having no demand on land area, absence of prolific and effective breeding and easy availability of fish when a person needs it are a few of the attractions of the technique. The studies indicate that it is desirable to have different meshes for the cages, such as, small meshed cages for rearing fry to fingerlings stages, and larger meshed cages for rearing fingerlings to table sized fishes. II' the meshes are small, the resistance will be more and less water wilt pass through. While feeding with powdered food material, because of brisk activity of feeding fish, a part of the feed appeared wasted. This can be easily overcome if we would resort to feeding fish with cheap pelleted feeds which will no doubt reduce wastage. Precaution has to be taken against damage of the net and thereby loss of fish and against poaching by unauthorised persons. In the present attempt has been demonstrated the possibility of utilizing locally available species of Tilapia for cage culture and obtaining moderately satisfactory growth rates.