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PubMed1999

Aluminum Polyphosphate Nanoparticles: Preparation, Particle Size Determination, and Microchemistry.

Admixture of aluminum nitrate, sodium polyphosphate, and ammonium hydroxide solutions yields stable dispersions of hydrated aluminum polyphosphate particles within a broad reagent concentration range. These particles are formed by liquid-liquid phase separation, for which a phase diagram was calculated using suitable models for concentrated electrolyte solutions. Particle effective diameters range from a few nanometers to many hundreds and are fractionated by centrifugation. Particle electrophoretic mobility is very low and the hydration degree is high ( approximately 80% v/v). Dry nanoparticles (1- to 15-nm diameter as observed by TEM) as well as particle aggregates are obtained by lyophilization. Element (P, Al, and Na) mapping by ESI-TEM shows that particle aggregates have a core-and-shell morphology, with a higher content of P in the aggregate core and a higher Na content at the outer shell. Copyright 1999 Academic Press.

Journal Article↗
PubMed1982

Aging, senile dementia, and the intralaminar microchemistry of cerebral cortex.

We compared the microchemical architecture or right frontal isocortex from patients with senile dementia and age-matched and younger controls. Neuronal connectivity within deep lamina of the cortical column (Brodmann area 9) tended to decline in normal aging and was profoundly depleted in senile dementia. In both aging and senile dementia, there was a significant 20% loss of total cells (neurons and glia) in cortical layers III to VI. In senile dementia, marked diminution of total ganglioside sialic acid per neuron and galactocerebroside per cell in the lower lamina far exceeded alterations associated with aging itself. This structural loss may imply deafferentation of the cortex, owing to loss of projections from subcortical areas such as nucleus basalis. Selective vulnerability of axodendritic arborization of neurons in lower lamina may be correlated to the impaired cognitive functions of senile dementia.

Adult↗
PubMed1991

Bedside diagnostic blood testing. Its accuracy, rapidity, and utility in blood conservation.

OBJECTIVE: --Bedside diagnostic testing utilizing microchemistry instruments potentially offers physicians the opportunity to evaluate urgent blood test results rapidly and reliably using only drops (250 microL) of whole blood. The use of microchemistry technology may also represent an essential component of a blood conservation program in acutely ill patients. We tested the hypothesis that a microchemistry instrument would have important advantages in the intensive care unit, including rapid turnaround time, decreased iatrogenic blood loss, and the provision of accurate analyte results. DESIGN: --One-year prospective, random-sample comparative study. SETTING: --A tertiary, acute care, 1000-bed university teaching hospital and a 450-bed university-affiliated community hospital. PATIENTS: --Blood samples from 850 patients were analyzed. RESULTS: --There were strong correlations (for pH, R2 = .97; PCO2, R2 = .97; PO2, R2 = .99; sodium, R2 = .83; potassium, R2 = .94; chloride, R2 = .90; glucose, R2 = .98; and hematocrit, R2 = .92) when paired samples were analyzed simultaneously in the laboratory comparing the microchemistry instrument and currently accepted clinical laboratory instruments. There were significant correlations (for pH, R2 = .91; PCO2, R2 = .94; PO2, R2 = .97; sodium, R2 = .91; potassium, R2 = .86; chloride, R2 = .91; ionized calcium, R2 = .82; glucose, R2 = .96; and hematocrit, R2 = .75) but increased variability occurred in some analyte results when samples were first analyzed in the intensive care unit (using the microchemistry instrument) and then transported to the laboratory in the routine fashion for "stat" determinations. CONCLUSIONS: --The multichannel microchemistry instrument provided accurate analyte determinations when compared with accepted clinical laboratory instruments. Significant savings of time and blood were also realized by the use of a microchemistry instrument in the intensive care unit. Bedside microchemistry may provide clinically important advantages in emergency departments, operating rooms, and neonatal, pediatric, coronary, adult medical and surgical, trauma, and burn intensive care units.

Blood Chemical Analysis↗
PubMed1988

Quantitative enzyme histochemistry in the brain.

Two main groups of quantitative methods are used in the brain to relate enzymatic processes to cellular structures, i.e. the methods of microchemistry and microscopic histochemistry. Microchemistry tries to quantify enzyme activities in very small brain regions by miniaturizing biochemical methods, whereas microscopic histochemistry applies staining procedures to tissue sections, preserving the structural relationship that is present in situ and giving topological information on the distribution of enzymes which is indispensable in structural heterogeneous tissue as is the brain. The present review deals preferentially with microscopic methods and, in particular, with scanning microphotometry (image plane scanning). Using this technique two measuring procedures can be applied for the quantification of enzyme activities, i.e. end-point and kinetic (continuous monitoring) measurements which are described in detail. Methods for the microphotometric demonstration of certain important dehydrogenases (isocitrate dehydrogenases, succinate dehydrogenase, NAD-linked malate dehydrogenase, glutamate dehydrogenase and glycerol 3-phosphate dehydrogenase), of cytochrome c oxidase, hexokinase and acetylcholinesterase are presented. These methods were adapted for giving optimal demonstration of enzyme activities in the rat hippocampus. The examples are given to illustrate the aptitude and possibilities of this technique in the quantification of enzymes in the complex matrix of the brain.

Brain↗
PubMed1981

The tadpole-shaped structures in human non-necrotizing granulomas.

We scrutinized the secondary lysosomal vesicle of the epitheloid cell from 9 patients whose biopsy specimens show multiple non-necrotizing granulomas by light microscopy (LM). In 4 of the 9 patients, we found tadpole-shaped structures (TSS) approximately tenfold larger than the size of the mycobacteriophage by transmission electron microscopy (TEM). In addition, we used a plasma etching method on epon-embedded tissue and successfully demonstrated the stereoscopic appearance of the TSS by scanning electron microscopy (SEM). The identified TSS were further analyzed with an X-ray energy dispersive spectrometer for their microchemistry. The TSS appeared to be integral structures by TEM and SEM and did not contain any nonbiologic elements when analyzed with the X-ray energy dispersive spectrometer. Their location as well as their morphologic features and microchemistry suggested that the TSS are a microorganism and related to the formation of the granulomas in our 4 patients.

Adult↗
PubMed2003

Optical detection in microscopic domains. 3. Confocal analysis of fluorescent amphiphilic molecules.

This series of papers demonstrates the feasibility of novel optical methodologies for microchemistry and analysis in aqueous samples of nano-, pico-, and femtoliter in volume. Not a conventional glass cuvette, but water-immiscible organic liquid, is used as the container for microscopic sample droplets in this approach. In part 1, absorption spectra of excellent quality were obtained and used for analysis from samples as small as a few tens of a micrometer in diameter. In part 2, an inert fluorescence marker as an internal standard was employed for indirectly detecting absorbing but nonfluorescent reagents in microsamples, employing inner filter effects. In this part 3, a third modality, confocal fluorescence microscopy, is added to the techniques being examined. A clearly visible emission ring emanating from an amphiphilic molecule, doxorubicin, at the sample boundary is demonstrated for the first time in "optically sliced" microdroplets. Relative intensity of this ring with respect to sample bulk can be used to study adsorption phenomena at liquid-liquid interfaces with proper calibrations for bulk and boundary. Quantitative separation of these two domains, a precondition to such calibrations, is also discussed.

Absorption↗
PubMedDate not supplied

[Scanning electron microscopy (SEM) and microanalysis (EDS) applied to the study of biomaterials for dental use. 6].

The study considers the use of scanning electron microscopy (SEM) and EDS microanalysis applied to the study of numerous mineral-based biomaterials of common use in odontostomatology. The products studied are the following: reabsorbable Dac Blu, non reabsorbable Dac Blu, non reabsorbable atomized Dac Blu, non rabsorbable fine Dac Blu, reabsorbable Biocoral 450, Calcitite 2040-12, Orthogel, Apagen, BTF 65, Calcitite 4060-2, Osprogel, Bio-Oss, Biostite, Ospro-vit, Merck Hydroxyapatite. By means of SEM it was possible to study the morphology and the microchemistry of the various biomaterials so as to have information about their physical and chemical characteristics, such as the crystalline form, the crystalline aggregations, the space dimensional distribution of the pores and check the possibilities of composition variability. All of these factors are fundamental to evaluate the functional biocompatibility of a biomaterial, once that its performance in a biological environment is known.

Biocompatible Materials↗
PubMedDate not supplied

High-precision continuous-flow isotope ratio mass spectrometry.

Although high-precision isotope determinations are routine in many areas of natural science, the instrument principles for their measurements have remained remarkably unchanged for four decades. The introduction of continuous-flow techniques to isotope ratio mass spectrometry (IRMS) instrumentation has precipitated a rapid expansion in capabilities for high-precision measurement of C, N, O, S, and H isotopes in the 1990s. Elemental analyzers, based on the flash combustion of solid organic samples, are interfaced to IRMS to facilitate routine C and N isotopic analysis of unprocessed samples. Gas/liquid equilibrators have automated O and H isotopic analysis of water in untreated aqueous fluids as complex as urine. Automated cryogenic concentrators permit analysis at part-per-million concentrations in environmental samples. Capillary gas chromatography interfaced to IRMS via on-line microchemistry facilitates compound-specific isotope analysis (CSIA) for purified organic analytes of 1 nmol of C, N, or O. GC-based CSIA for hydrogen and liquid chromatography-based interfaces to IRMS have both been demonstrated, and continuing progress promises to bring these advances to routine use. Automated position-specific isotope analysis (PSIA) using noncatalytic pyrolysis has been shown to produce fragments without appreciable carbon scrambling or major isotopic fractionation, and shows great promise for intramolecular isotope ratio analysis. Finally, IRMS notation and useful elementary isotopic relationships derived from the fundamental mass balance equation are presented.

Animals↗
PubMed2002

Interactions between MC3T3-E1 cells and textured Ti6Al4V surfaces.

This paper presents the results of an experimental study of the interactions between MC3T3-E1 (mouse calvarian) cells and textured Ti6Al4V surfaces, including surfaces produced by laser microgrooving; blasting with alumina particles; and polishing. The multiscale interactions between MC3T3-E1 cells and these textured surfaces are studied using a combination of optical scanning transmission electron microscopy and atomic force microscopy. The potential cytotoxic effects of microchemistry on cell-surface interactions also are considered in studies of cell spreading and orientation over 9-day periods. These studies show that cells on microgrooved Ti6Al4V geometries that are 8 or 12 microm deep undergo contact guidance and limited cell spreading. Similar contact guidance is observed on the surfaces of diamond-polished surfaces on which nanoscale grooves are formed due to the scratching that occurs during polishing. In contrast, random cell orientations are observed on alumina-blasted Ti6Al4V surfaces. The possible effects of surface topography are discussed for scar-tissue formation and improved cell-surface integration.

Alloys↗
PubMed1981

The use of a hidden metal-capture reagent for the measurement of Na+--K+-ATPase activity: a new concept in cytochemistry.

The original lead-trapping method for demonstrating Na+--K+-ATPase activity was discredited because of the effect that lead ions can have on the substrate and on the enzyme. Current methods, that measure this activity by the related K+-dependent phosphatase activity, do not appear to measure activity that is known, from microchemistry, to occur in proximal convoluted tubules. The disadvantages of using lead appear to have been overcome by the use of a new reagent in which the lead is complexed with ammonium citrate ions; phosphate, liberated enzymatically, successfully competes with these ions. The activities of total ATPase and of the ouabain sensitive Na+--K+-ATPase have been measured in three regions of the nephron in the guinea-pig and in the rat. The relative activities found, by this method, in the different regions of the latter, appear to be comparable with results found by others, using microchemical methods applied to isolated regions of the nephron.

Animals↗
PubMed1991

Assessment of lysosomal function by quantitative histochemical and cytochemical methods.

Quantitative histochemistry and cytochemistry enables a direct link to be made between metabolic functions such as the activity of lysosomal enzymes and the morphology of a tissue or a type of cell. Several approaches exist such as microchemistry based on (bio)chemical analysis of a single cell or a small piece of tissue dissected from a freeze-dried section. This technique has been routinely used for prenatal diagnosis of inherited enzyme defects and especially of lysosomal storage diseases. Other approaches are cytofluorometry or cytophotometry, which are based on the principle that a fluorescent or coloured final reaction product is precipitated at the site of the enzyme. The amount of final reaction product is analysed per cell or per unit volume of tissue using either a microscope cytofluorometer or flow cytometer for fluorescence measurements or an image analysing system or scanning and integrating cytophotometer for absorbance measurements. In principle, fluorescence methods are to be preferred over chromogenic methods because they are more sensitive and enable multiparameter analysis. However, only a limited number of fluorogenic methods are at hand that give a final reaction product which is sufficiently water-insoluble to guarantee good localisation. The best results have been obtained with methods based on naphthol AS-TR derivatives and with methods for the demonstration of protease activity using methoxynaphthylamine derivatives as substrates and 5'-nitrosalicylaldehyde as coupling reagent. Chromogenic methods are far better with respect to localisation properties and, therefore, most commonly used for quantitative histochemical analysis of lysosomal enzyme activities.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytophotometry↗
PubMed1977

Pancreatic islets subjected to different concentrations of glucose in vitro. A study with special regard to mitochondrial changes.

Isolated pancreatic islets of mice and gerbils were cultured for 6 days at low (2mM) or high (20mM) concentrations of glucose after which they were studied using qualitative and quantitative electron microscopy, histo- and microchemistry, and X-ray microanalysis. Compared with the islets cultured at high glucose, those subjected to low glucose exhibited enhanced succinate dehydrogenase activity, a decreased content of adenosine triphosphate, and an increased volume of B-cell mitochondria which often were rounded or oval.

Adenosine Triphosphate↗
PubMed1999

Versican in human fetal skin development.

The extracellular matrix of human fetal skin differs substantially from that of adult skin. Fetal skin contains sparse amounts of fibrillar collagen enmeshed in a highly hydrated amorphous matrix composed of hyaluronan and sulfated proteoglycans. Both fetal and adult skin contain two major interstitial proteoglycans that are extracted by chaotrophic agents and detergents. These are the large chondroitin sulfate proteoglycan versican and the small dermatan sulfate proteoglycan decorin. For this study, proteoglycans extracted from fetal and adult skin were compared on Western blots to determine the relative amounts of versican. Decorin present in the same samples provided an internal standard for these studies. Fetal skin differed from adult skin in that it contained a significantly higher proportion of versican than did adult skin. Immunohistochemical studies compared early-fetal with mid-fetal skin and found that versican was a significant component of the interstitial extracellular matrix at both of these stages of skin development. However, by the mid-fetal period, interstitial versican became restricted to the upper half of the dermis, although versican also continued to be highly expressed around hair follicles, glands, and vasculature in the lower half of the dermis. Fetal skin extracts differed from an adult skin extract by the presence of a 66-kDa protein immunologically related to versican and by the absence of a 17-kDa core protein of a proteoglycan related to decorin. Both of these molecular species may represent degradation products of their respective proteoglycans. Monoclonal antibodies which detect epitopes in native chondroitin sulfate glycosaminoglycan chains recognized versican extracted from fetal skin. However, the tissue distribution of these antigens did not entirely conform to that for versican core protein, suggesting that versican in different regions of the skin may be substituted with glycosaminoglycan chains with different microchemistries. The results of these studies indicate that human fetal skin is structurally different from adult skin in terms of both the distribution and the composition of the large, aggregating chondroitin sulfate proteoglycan versican.

Adult↗