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Pinocytotic function of the membrane vesicles, or caveolae, in heart muscle cells.

The structure of membrane vesicles present in the sinus node, the atrium, and the ventricular cells of rabbits and mice was investigated. The results described in this study support the idea that membrane vesicles are not pinocytotic vesicles, but structural components of the membrane that increase the sarcolemmal surface area. That is the reason why we prefer to call them caveolae. Why these caveolae are much more abundant in sinus node cells and in Purkinje fibers, as shown by Mobley and Page (1972), than in working myocardium is still unknown.

Animals↗

[Practical staining method: T-OPA triple stain].

The following procedure has proven to be successful as routine trichrome stain on paraffin embedded material: 1) Mayer's hemalum for 10 min, followed by running tap water wash; 2) staining in 1% Orange G in 1% acqueous PTA for 5 min and rinsing a few seconds in distilled water; 3) Aniline blue 1% acqueous for 5 min, followed by few seconds distilled water wash. Dehidratation in ethanol, or by blotting followed by t-buthanol or 1:3 terpineol-xylene, clearing and mounting, completed the procedure.

Acridine Orange↗

[Evaluation of the accuracy of precipitation methods for HDL cholesterol determination].

Several methods for HDL-Cholesterol determination have been proposed; the over-floating cholesterol evaluation after non-HDL lipoproteins precipitation by polyanions with bivalent cations or neutral polymers is practical and not expensive, but it's accuracy and precision must be strictly control led. The Authors, after a review and a quality-check of the most used methods, conclude that the best accuracy for HDL-Cholesterol determination is provided by Dextransulphate-Mg-chloride and Phosphotungstate-Mg-chloride at pH 7,5.

Chemical Phenomena↗

Morphologic unity of Paneth cells.

Paneth cells occurring in intestinal and extraintestinal sites were studied using various light microscopic techniques, including standard hematoxylineosin (HE)-stained histologic sections; a number of histochemical reactions, including the phosphotungstic acid-hematoxylin (PTAH) method; and immunohistochemical labeling for the presence of lysozyme using the peroxidase-antiperoxidase (PAP) method. Cells, identified on HE as Paneth cells, uniformly stained with PTAH and PAP-lysozyme. Further, all of the cases demonstrating epithelial cell lysozyme showed eosinophilic cytoplasmic granules, typical of Paneth cells, on HE sections. Lysozyme was demonstrated in benign and malignant ovarian tumors, as well as in a variety of ectopic intestinal sites and benign and malignant intestinal tumors, supporting the concept that the elaboration of lysozyme is a nonspecific feature of intestinal-cell differentiation and is not a response to a specific stimulus.

Cystadenocarcinoma↗

Comparison of the Du Pont aca and Dow methods for determination of high-density lipoprotein cholesterol.

We compared the Du Pont aca (phosphotungstate-enzymic cholesterol) and the Dow (dextran sulfate/Mg2+-enzymic cholesterol) methods for the determination of high-density lipoprotein cholesterol (HDLC) and total cholesterol in serum from 113 patients. The aca results for both total cholesterol and HDLC were significantly greater (p less than 0.0001) than the Dow results, the aca method overestimating the HDLC concentration (mean recovery 107.2% in serum samples with values assigned by the Centers for Disease Control). The precision of the aca method for HDLC was essentially the same as that of the Dow method. Bilirubin (up to 0.17 g/L), hemoglobin (up to 4 g/L), and slight lipemia (triglycerides up to 5.4 g/L) did not interfere with the aca method.

Autoanalysis↗

Glycoproteins of bovine epididymal spermatozoa--a cytochemical study.

Modifications in bull sperm plasmamembrane during epididymal passage were investigated by the use of four different lectins: Concanavalin A (Con A); Ricinus communis I (RCA1); Wheat germ agglutinin (WGA); Ulex europaeus agglutinin I (UEA1). During sperm passage from caput to cauda epididymidis agglutination by RCA1 and WGA distinctly increased. Similar but somewhat less pronounced difference in the agglutinability was found for Con A. No agglutination was observed with UEA1. Ultrastructural examination of Con A binding sites on sperm plasma membrane with a Con A-horseradish peroxidase-gold technique (Con A-HRP-G) revealed a significant increase in the number of gold granules on the sperm tails during the epididymal passage of spermatozoa. No change in WGA-binding sites was observed between caput and cauda spermatozoa using a WGA-peroxidase method.

Agglutination↗

Precipitation of apo E-containing lipoproteins by precipitation reagents for apolipoprotein B.

We measured the solubility of apolipoprotein E (apo E) after precipitation, with heparin-Mn2+ or dextran sulfate-Mg2+, of lipoproteins containing apo B. Data from 46 randomly selected subjects suggest that apo E is readily precipitated by dextran sulfate-Mg2+, but that heparin-Mn2+ preferentially precipitates apo E associated with apo B-containing lipoproteins while leaving the apo E-containing fraction of high-density lipoproteins (HDL) in solution. In a more detailed analysis of three subjects, we measured the lipoprotein association of apo E by column chromatography on agarose beads, before and after its precipitation from plasma. This study confirmed the preferential solubility of apo E associated with HDL lipoproteins. Using plasma from two normolipidemic subjects, we maintained the heparin concentration at 1.30 g/L and varied the manganese concentration from 9.2 to 184 mmol/L. A 46 mmol/L concentration best separated apo E-containing HDL from apo B-containing lipoproteins. Thus, at these final concentrations, heparin-Mn2+ appears to precipitate the apo E associated with apo B-containing lipoproteins, leaving soluble most of the apo E associated with lipoproteins of HDL size.

Apolipoprotein A-I↗

Studies of the accuracy of measurements of serum high density lipoprotein cholesterol levels.

Titration of serum lipoproteins with three different precipitants--heparin-MnCl2 92 mmol.L-1, dextran sulfate-MgSO4, and phosphotungstate-MgCl2--revealed gross errors in the high density lipoprotein cholesterol (HDL-C) levels measured in some hyperlipidemic sera. Dilution studies of eight normal sera with the phosphotungstate method revealed that the most reliable estimate of the HDL-C level was the level measured in the least diluted aliquot that yielded a good precipitate. With a heparin-MnCl2 kit, serial dilution of a lipemic serum demonstrated a requirement for a blank correction. Establishing an accurate estimate of the HDL-C level sometimes requires knowledge of the titration curve of the lipoprotein precipitant being used, a blank correction, and data obtained from dilution studies (or studies with increased precipitant levels) on the serum in question.

Cholesterol, HDL↗

Quantification of high-density-lipoprotein cholesterol by precipitation with phosphotungstic acid/MgCl2.

We evaluated the use of a modified phosphotungstic acid/MgCl2 precipitation procedure for the precipitation of apolipoprotein B-containing lipoproteins. Precipitation of these lipoproteins [very-low- and low-density lipoproteins, and lipoprotein (a)] is complete, with negligible coprecipitation of high-density lipoprotein subfractions (HDL1, HDL2, HDL3), even in hypertriglyceridemic sera. In comparison with ultracentrifugation, the precipitation method yields, on the average, values that are 0.17 mmol/L lower for cholesterol values but almost identical for apolipoprotein A-I and phosphatidylcholine. Looking for delta 3,5-cholestadiene formed from cholesterol in the precipitation residue, we used "high-performance" liquid chromatography and "high-performance" thin-layer chromatography and found none.

Apolipoproteins↗

Microcystic meningioma. Clinicopathologic report of eight cases.

We report herein eight cases of microcystic meningiomas. This unusual variant was originally described by Masson, who labeled it "humid." Grossly, tumors were usually soft, with occasionally a glistening cut surface. Two tumors were cystic and three invaded bone structures. By light microscopy, tumor cells were stellate, with long processes delimitating clear spaces. Whorls were found in all tumors. Intracytoplasmic glycogen, fatty droplets, and phosphotungstic acid-hematoxylin-positive fibrils were sometimes seen. Tumor cells, however, were all negative with the peroxidase-antiperoxidase technique for glial fibrillary acidic protein. Mitoses, hemorrhages, and necrosis were absent. By electron microscopy, interdigitations of cytoplasmic processes and desmosomes were found. The large extracellular spaces were filled with a finely granular substance intermingled with fibrils and collagenic fibers. The clinical and morphological findings point toward a benign course. We also discuss the differential diagnosis and the pathogenesis.

Adolescent↗

Production of skeletal muscle elements by cell lines derived from neoplastic rat mammary epithelial stem cells.

Single-cell-cloned cell lines intermediate in morphology between the cuboidal epithelial and fully elongated myoepithelial-like cells have been isolated from the single-cell-cloned epithelial stem cell lines Rama 25 and Rama 37 originally obtained from dimethylbenz(a)anthracene-induced mammary tumors from Sprague-Dawley and Wistar-Furth rats, respectively. These are designated Rama 25-l1, Rama 25-l2, Rama 25-l4 (Sprague-Dawley) and Rama 50-55, Rama 59, and Rama 60 (Wistar-Furth), respectively. When growing as tumors in nude mice or syngeneic Wistar-Furth rats, respectively, many of the newly cloned cell lines give rise to spindle and giant, multinucleated cells which stain immunocytochemically with antisera to myoglobin and myosin and contain longitudinal fibrils, some of which contain phosphotungstic acid-hematoxylin-staining cross-striations. Ultrastructural analysis demonstrates the presence of A-, l-, and H-bands and Z-discs and the hexagonal arrangement of thick and thin filaments characteristic of skeletal muscle. Similar results are obtained with selected cloned cell lines growing on floating collagen gels in vitro. Thus, a developmentally committed mammary epithelial cell can give rise, under suitable conditions, to a well-differentiated mesenchymal lineage, that of skeletal muscle. It is suggested that such cells may be responsible for the generation of the well-differentiated mesenchymal elements seen in the mixed (epithelial and myoepithelial) tumors of glandular origin.

Animals↗

[Accuracy of HDL-cholesterol and HDL-apolipoprotein A-I determinations after phosphotungstate/MgCl2-precipitation of apolipoprotein B-containing lipoproteins (author's transl)].

With optimal concentrations of reagents a complete precipitation of apolipoprotein B-containing lipoproteins and only a small coprecipitation of HDL can be achieved using the phosphotungstate/MgCl2 precipitation method. The accuracy of the precipitation is not influenced by triglyceride concentrations less than 4.0 mmol/l. Similarly, in most hypertriglyceridaemic sera (greater than 4.0 mmol/l) a complete precipitation of apolipoprotein B-containing lipoproteins and a complete recovery of HDL can be observed. In addition to the HDL cholesterol determination, apolipoprotein A-I was determined by kinetic nephelometry. In hypertriglyceridaemic sera (greater than 4.56 mmol/l) precipitation of apolipoprotein B-containing lipoproteins must be carried out prior to the nephelometric determination of apolipoprotein A-I.

Aging↗

[The enzymatic analysis of sphingomyelin in HDL (author's transl)].

A simple method is described for the enzymatic determination of sphingomyelin in the apolipoprotein B-free supernatants prepared by precipitation of blood sera with phosphotungstate/MgCl2. The analysis is based on the enzymatic hydrolysis of sphingomyelin, by sphingomyelinase from B. cereus, into phosphorylcholine and N-acylsphingosine, and subsequent hydrolysis of phosphorylcholine by alkaline phosphatase. The choline formed is determined by choline kinase in an optical test. The results from this method were in good agreement with those obtained by the conventional chemical sphingomyelin determination. Furthermore, there was a good correlation between the sphingomyelin concentrations obtained from the HDL fractions isolated by ultracentrifugation (1.063-1.21 kg/l) and those obtained from the apolipoprotein B free supernatants after phosphotungstate/mgCl2 precipitation of sera.

Bacillus cereus↗