acid-phosphatase and copper-phthalocyanine

In this work, a novel sensor based on immobilised copper phthalocyanine, 2,9,16,23-tetracarboxylic acid-polyacrylamide (Cu(II)TC Pc-PAA) was developed for determination of acid phosphatase (ACP) levels in nanomolar quantities. Detection was based on the measurement of enzymatically generated phosphate, with initial studies focused on phosphate detection at a Cu(II)TC Pc-PAA modified screen-printed gold transducer. The sensor was characterised in relation to operational performance (pH, response time, stability, linearity, and sensitivity) and common anionic interferents (nitrate, sulphate, chloride, and perchlorate). The functionalised surface also facilitated rapid detection of the enzyme bi-product 2-naphthol over the range 5-3000 μM. Quantitation of ACP was demonstrated, realising a linear response range of 0.5-20 nM and LOD of 0.5 nM, which is within the clinical range for this prostate cancer biomarker.

Topics: Acid Phosphatase; Biomarkers, Tumor; Electrochemistry; Electrodes; Gold; Humans; Hydrogen-Ion Concentration; Indoles; Limit of Detection; Male; Organometallic Compounds; Perchlorates; Printing; Prostatic Neoplasms; Surface Properties; Time Factors; Transducers

The space between the oesophageal basal and prickle epithelial cells appears empty by standard ultrastructural preparative techniques. Fixation of human oesophageal biopsies with a variety of agents, including tannic acid, glutaraldehyde-lysine, cetylpyridinium chloride and Ruthenium Red shows that this space is filled with mucosubstances, some free, some attached to the cells as a glycocalyx. There is evidence that this material is secreted constitutively by the basal and prickle cells. This secretion may be changed or blocked by incubating oesophageal biopsies in the presence of colchicine or dinitrophenol. Incubation at 16 degrees C has the same effect. Absorption from the intercellular space may be followed using the fluid phase marker, horseradish peroxidase. Early endosomes may also be shown by their acid phosphatase activity. Incubation of biopsies at 20-22 degrees C allows early endosomes to accumulate material, but not pass it on the late endosomes.

Topics: Absorption; Acid Phosphatase; Adult; Aged; Aged, 80 and over; Biopsy; Epithelial Cells; Epithelium; Esophagus; Female; Glutaral; Glycoconjugates; Glycoproteins; Golgi Apparatus; Humans; Indoles; Lysine; Male; Middle Aged; Mucoproteins; Organometallic Compounds; Polysaccharides; Secretory Rate; Staining and Labeling; Tissue Fixation

The pulmonary intravascular macrophages (PIMs) of the ponies possess all the characteristics of a fully differentiated resident macrophage, which forms adhesive plaques with the capillary-endothelial cells. In addition, it has a unique surface coat which does not conform to the concept of a carbohydrate rich membrane bound glycocalyx generally associated with macrophages. We studied the responses of these cells especially in the context of its globular surface coat to multiple doses of MB intravenously: 0.2 ml/kg body copper tracer substance. The ponies were treated with three doses of MB intravenously: 0.2 ml/kg body weight on day 0, 0.1 ml/kg at 48 h and a 0.05 ml/kg at 96 h after the first dose. The examination of lung tissue at the ultrastructural level revealed internalization of MB following complexing with the surface coat globules along the entire cell surface. The cell surface was extensively ruffled in the form of several lamellipods. The MB-globule complex was internalized mostly at the coated pits whereas concurrent internalization of nascent globules of the coat took place at the ruffled surface. In the former, the internalized material was seen in small, medium and large phagolysosomes which stained intensely for acid phosphatase, whereas in latter case enzyme activity was entirely absent in all the nascent globules carrying endosomes. Simultaneously there was hypertrophy of the Golgi complex, which showed intense staining for acid phosphatase. Relative to the Golgi response, acid phosphatase-positive tubular lysosomes in close association with extensive bundles of microtubules were conspicuous, thus adding to the battery of cell organelles of secretory pathway.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Acid Phosphatase; Animals; Cell Communication; Cell Membrane; Dose-Response Relationship, Drug; Endothelium, Vascular; Female; Golgi Apparatus; Histocytochemistry; Horses; Indoles; Injections, Intravenous; Lysosomes; Macrophages, Alveolar; Male; Microscopy, Electron; Microtubules; Organometallic Compounds

We previously reported that the pulmonary intravascular macrophages (PIMs) of sheep, goat, and calf lung contained a heparin and a lipolytic lipase sensitive surface coat by using tannic acid as a component of paraformaldehyde-glutaraldehyde-based fixative. The implication of this sensitivity was that the surface coat was predominantly comprised of lipoprotein-like substance. In this study we report that monastral blue (MB) used as a vascular tracer interacted with the coat globules and lost its original particulate appearance. Its precise localization in the PIMs was in combination with altered macromolecules of the surface coat in the form of lipid droplets, which conformed to the conventional view of neutral lipids. In contrast, pigment particles examined in their native state resembled metallic particles as electron-dense elliptical rods. The lipid droplets were subsequently internalized through endocytic route and found their access into the lysosomal compartments of PIMs at the electron microscopic level. Lamellar bodies (LLBs) arose from the lysosomal matrix after the entry of lipid droplets in the secondary lysosomes. Acid phosphatase activity was located in secondary lysosomes as well as in endosomes. These observations suggest that coat granules of the PIMs acted as a carrier of exogenous MB particles to deliver the complex to the lysosomal compartment where partial digestion lead to the formation of lamellar bodies. The implications of MB (cationic dye) as a vascular tracer for studying phagocytic index of PIMs in the light of their coat and the rapid development of LLBs are discussed. It is proposed that MB by initially combining with the surface coat provokes mobilization of intracellular lipid pools. In this way metabolism of vasoactive lipid in the PIMs is stimulated to influence the dynamics of pulmonary circulation in the calves.

Topics: Acid Phosphatase; Animals; Blood Vessels; Cattle; Coloring Agents; Endocytosis; Histocytochemistry; Indoles; Kupffer Cells; Lysosomes; Macrophages; Male; Microscopy, Electron; Organometallic Compounds; Phagocytosis; Pulmonary Circulation; Surface Properties

The pulmonary intravascular macrophages (PIMs) have been described in several species of animals. This study demonstrates for the first time that the equine lung has PIMs as resident phagocytes in its microvasculature. Their salient features such as globular surface coat, structures of the endocytic pathway, and related cell organelles closely resemble those of the calf, goat, and sheep. The exquisite organization of the coat globules in the form of a linear chain was structurally similar to the lipolytic lipase and the heparin-sensitive globular coat from PIMs of calf, goat, and sheep. Monastral blue (MB) when employed as a tracer to assess the phagocytic properties of equine PIMs induced similar modification of the globules of the coat into lipid droplets, reminiscent of neutral lipids. Lipids droplets (modified coat globules) were delivered into acid phosphatase-positive endosomes and lysosomes. Concurrently, the unaltered globules of the coat, probably internalized via fluid-phase constitutive pinocytoses, followed a different endocytic pathway. Large-scale platelet uptake by the PIMs was observed with thrombocytopenia in MB-treated ponies. The possible significance of hypothetical LDL-coat and the endocytic organelles as equivalents of synthetic apparatus of vasoactive lipids in the PIMs of horse needs to be assessed in future studies.

Topics: Acid Phosphatase; Animals; Female; Horses; Indoles; Lung; Lysosomes; Macrophages, Alveolar; Male; Microscopy, Electron; Organometallic Compounds; Phagocytosis