RESUMEN
Morphometrical data on the intrahepatic arterial wall were studied over a wide range of vascular diameters (100-1708 microns) in one human liver. The liver tissue containing Araldite-filled blood vessels was embedded in the water-soluble glycolmethacrylate (GMA). The calculation of the correction factor for the morphometric data obtained from 3 microns sections is described. The influence of formaldehyde fixation on the volume of the liver proves to be negligible. During dehydration a linear shrinkage of about 3% occurs. After infiltration and embedding in GMA only 1% of this shrinkage remains. During the drying phase about 4.3% further linear stretching occurs. No significant "residual compression factor" was found. Araldite plays a significant role in the retention of the dimensions of the liver specimen during histologic processing, while the dimensional changes of the Araldite itself are negligible. A positive linear correlation was found between the media thickness and the radius of the vessel. The physiological consequences are discussed. It is concluded that in the morphometric analysis of the arterial wall it is essential to apply a standardized procedure in histologic processing and in the measuring of the inner vascular diameter. The advantages of our method, in which blood vessels are perfused with Araldite under physiological pressure, are discussed.
Asunto(s)
Arteria Hepática/anatomía & histología , Adulto , Resinas Epoxi , Humanos , Hígado/anatomía & histología , Masculino , Músculo Liso Vascular/anatomía & histología , Anhídridos Ftálicos , Análisis de RegresiónRESUMEN
This paper describes the morphometric changes liver samples undergo in the course of fixation, dehydration, infiltration and embedding in different mixtures of water-soluble plastics. The plastics used were: three different mixtures of glycolmethacrylate (GMA) and the commercially available material JB4. Buffered formaldehyde fixation did not produce significant morphometric changes in the liver specimens. Dehydration obviously affects the volume of the liver specimen (linear shrinkage about 9.3%). The dehydration is followed by an infiltration phase. During this phase a slight swelling (linear, 2-5%) occurs. The final polymerization of the plastic resulted in a further linear shrinkage of 1-2%. The influence of different technical factors on the stretching of sections of pure plastic and of plastic embedded liver appeared to be considerable. The difference in stretching between 2 and 3 micron sections has been studied. A significant influence of temperature upon section stretching was noted. Sections of all plastic mixtures stretched at a temperature of 293 K showed 3% more linear stretch than at a temperature of 333 K. Differences between the four plastic mixtures are discussed. Correction factors must be used in morphometrical and stereological investigations (see for review: Weibel, 1979). It was concluded that in the application of water-soluble plastics as embedding media it is essential to apply a standardized procedure, particularly in the cutting and stretching phase.
Asunto(s)
Hígado/citología , Animales , Técnicas Histológicas , Plásticos , Solubilidad , Porcinos , AguaRESUMEN
Male and female rats with two permanently indwelling intravenous catheters were infused for 2 hours with ovine prolactin. During equilibrium conditions the effects of intravenously injected L-DOPA and benzerazide (a blocker of dopa-decarboxylase) on steady state levels of ovine prolactin were measured. A dose of 4.5 mg L-DOPA per 100 gr body weight (b.w.) caused a transient increase of plasma ovine prolactin. A dose of 0.3 mg L-DOPA/100 gr b.w. had no effect, neither in males nor in females, while benzerazide (20 mg/100 gr b.w.) had only a slight effect. The experiments suggest that L-DOPA does not affect the peripheral uptake of prolactin from the plasma.
Asunto(s)
Levodopa/farmacología , Prolactina/sangre , Animales , Benserazida/farmacología , Femenino , Lactancia , Levodopa/administración & dosificación , Masculino , Embarazo , Prolactina/metabolismo , Ratas , OvinosRESUMEN
In this paper a technique is described, using Araldite CY 223 and hardener HY 2967 as injection material, for preparing corrosion casts or histological sections. The plastic has a viscosity (at 39-40 degrees C) similar to that of blood, a gelling time of approximately 17 min (at 40 degrees C), and an exothermic transition energy of delta H = 80.28 +/- 3.20 cal/gm. The influence of the plastic on the tissue is discussed. The histological sectioning of fixed tissue containing Araldite-filled blood vessels after embedding in 2-hydroxyethyl-methacrylate (GMA) is described. When using GMA in a modification of the mixtures of Ruddell (1967) and Sims (1974), methylbenzoate is recommended as an intermedium in order to obtain a more uniform infiltration and reproducible section thickness. At the same time methylbenzoate is recommended as a storing fluid. Sections of 2-3 micrometers afford satisfying morphologic and morphometric results. This method allows various arterial wall dimensions to be measured easily, and provides a suitable means to compare histometric values with SEM data derived from corrosion casts.
Asunto(s)
Vasos Sanguíneos/anatomía & histología , Hígado/irrigación sanguínea , Anhídridos Ftálicos , Plásticos , Fenómenos Químicos , Química , Resinas Epoxi , Estudios de Evaluación como Asunto , Humanos , Métodos , Microscopía Electrónica de Rastreo , Modelos AnatómicosRESUMEN
In a comparative study, the enzyme-linked immunosorbent assay, using peroxidase labeled anti-rat immunoglobulin M and immunoglobulin G, and the passive hemagglutination test were applied to determine the primary and secondary antibody response to lipopolysaccharide and tetanus toxoid in rats. In the enzyme-linked immunosorbent assay, the antigens were bound to the wells of polystyrene microplates, tetanus toxoid directly, and lipopolysaccharide after complexing it with methylated bovine serum albumin. After incubation with dilutions of the rat sera, the amount of antibody bound to the solid phase was quantified by means of peroxidase-labeled anti-immunoglobulin. The specificity of the enzyme immunoassay was tested by absorption of the sera with their respective antigens. The enzyme-linked immunosorbent assay proved to be more sensitive than the hemagglutination reaction, except when titers were determined during the secondary response to tetanus toxoid. Besides its specificity and sensitivity, the enzyme-linked immunosorbent assay is a convenient method for measuring both immunoglobulin M and immunoglobulin G antibodies. At low serum dilutions of lipopolysaccharide antisera, inhibition of the reaction in the enzyme-linked immunosorbent assay occurred. This phenomenon could be prevented by heating the sera at 56 degrees C for 30 min. Lipopolysaccharide was immunogenic in rats over an extremely wide dose range (from 10 pg to 1 mg); the optimal immunogenic dose of lipopolysaccharide for young adult rats was 0.1 to 1,000 mug when administered intravenously, and that of tetanus toxoid was 5 to 10 lines of flocculation, as determined by the Ramon flocculation test.
Asunto(s)
Anticuerpos Antibacterianos/análisis , Ensayo de Inmunoadsorción Enzimática , Pruebas de Hemaglutinación , Técnicas para Inmunoenzimas , Lipopolisacáridos/inmunología , Antitoxina Tetánica/análisis , Toxoide Tetánico/inmunología , Animales , Inmunoglobulina G/análisis , Inmunoglobulina M/análisis , Ratas , Factores de TiempoRESUMEN
The development of Barbus conchonius is described with special attention to the differentiation of the gut. Amine precursor uptake and decarboxylation (APUD) are present in enteroendocrine cells during development, whereas these processes are lacking in adult specimens. The first APUD cells originate on the fourth day of development in the anterior part of the gut and on the fifth day in the caudal areas. The APUD facility of the cells disappears within 2 days, and after the 6th day APUD cells can no longer be distinguished in the intestinal epithelium. The first APUD cells were obsserved when four types of enteroendocrine cells were recognized with the electron microscope. These enteroendocrine cells contain granules of different electron densities, and microtubules and cilia can be observed. Some enteroendocrine-like cells are found below the basement membrane of the intestinal epithelium, indicating a possible extra-endodermal origin. APUD cells, except melanoblasts, have not been found migrating from the neural crest in ventral direction. The origin of the enteroendocrine cells of B. conchonius is discussed.