The staining properties of thiazolylazoalkylaminophenol derivatives in histochemical staining of cadmium

In the continuous research for a more sensitive chelator for cadmium, thiazolylazoalkylaminophenol derivatives were synthesized. The staining sensitivity for cadmium and molar absorptivity of their cadmium chelates were examined. Though the molar absorptivity of bromothiazolylazodiethylaminophenol(Br-TADA)-cadmium chelate demonstrated an order of 10(5), the staining sensitivity of Br-TADA was unexpectedly low. The lowering of the sensitivity of Br-TADA may be attributed to alkaline hydrolysis and a decrease in the stability constant of the cadmium chelate.     

Morphine sulphate induced histopathological and histochemical changes in the rat liver

In this study, the histopathological and histochemical changes due to chronic usage of morphine sulphate in liver were assessed in rats with both light and electron microscopes. Twenty male albino rats (Rattus norvegicus) (130-150 g) were included and divided into four groups. Normal saline (5 ml) was given orally as placebo in the control group (N = 5). Morphine groups (N = 5) received morphine orally at a single dose of 5 ml/kg/day for 10, 20 and 30 days (groups II, III and IV), respectively. Liver specimens from all groups were evaluated for histopathological and histochemical changes. Light microscopy revealed severe centrilobular congestion, portal fibrosis with bile ductal proliferation and an increased inflammatory infiltration and focal parenchymal necrosis. Histochemical study revealed a progressive depletion of general carbohydrates and an increase in total protein contents. These changes were confirmed at ultrastructural level, including the presence of accumulated lipid in the hepatocytes; deposits of a collagen-like fibrous material were seen in the space of Disse and a reduction in the number of endothelial cell fenestrations. Our findings pointed out the risk of increased lipid fibrosis and hepatic damage due to long-term use of morphine. Although opioids are reported to be effective in pain management, their toxic effects should be kept in mind during chronic usage.     

The histochemical demonstration of aniline hydroxylase activity in rat liver

Synopsis A procedure is described for the histochemical demonstration of aniline hydroxylase activity in cryostat sections of rat liver. Tissue sections are incubated in a medium containing aniline; thep-aminophenol formed as a result of enzymatic action is coupledin situ with Fast Blue RR. The staining reaction is found to be confined to the cytoplasm of the hepatocytes. Confirmatory tests for true enzymatic staining reaction include the incubation of sections in medium from which aniline is omitted, and under conditions of enzyme inhibibition. A method for the quantitation of the histochemical staining reaction is also described.The histochemical reactions have been investigated on rat livers subjected to conditions eliciting microsomal enzyme stimulation and inhibition, bothin vitro andin vivo. A close correlation was found between the staining reactions observed and the results of the quantitative histochemical method and the biochemical estimations of aniline hydroxylase activity in liver microsomal fractions obtained by differential centrifugation.     

Atrial specific granules of the rat heart: light microscopic staining and histochemical reactions.

An investigation was carried out to determine general staining and histochemical properties of rat atrial specific granules. It was found that these granules may be demonstrated using aldehyde fuchsin after pretreatments which involve oxidation or thiosulfation. This new way of demonstrating atrial granules is compared to other staining methods in terms of sensitivity and selectivity as well as to the nature of reactive groups that may be involved in the staining reactions. No lipid or carbohydrate were detected histochemically. Overall assessment of reactions suggests that atrial granules are a site of storage for a protein or polypeptide. Some of the tests indicate that these may contain tryptophan and sulfur-containing amino acids.     

Feulgen staining of rat liver fixed in different fixatives.

In the present study rat liver pieces fixed in 1) 10 per cent buffered neutral formalin, 2) 4 per cent glutaraldehyde, 3) Heidenhain's-Susa fixative and 4) Flemming's fluid, and following hydrolysis in 1-0 N HC1 at 60degreesC for varying time periods have been stained with the UV Feulgen procedure. The results of this study reveal that following hydrolysis for different time periods the tissue material fixed in formalin show the same staining pattern as those fixed in glutaraldehyde. The material fixed in Heidenhain's-Susa displays an intense Feulgen staining after two different times of hydrolysis, and that fixed in Flemming's fluid shows particular staining intensity for a prolonged time period thus indicating better preservation of DNA than in the materials fixed in the other three fixtatives.     

Phospholipid interactions in rat liver nuclear matrix

Rat liver nuclear matrix has been isolated by salt extraction and nuclease digestion of nuclei. Under the electron microscope, the matrix appears as a spongelike network joined by thinner fibrils. Biochemical analysis shows a high protein content and low amounts of nucleic acid and phospholipid. Treatment of the matrix with phospholipase C results in a release of most of the nucleic acid, and a disappearance of the fibrils, however the appearance of the matrix is largely unaffected. It seems likely that phospholipids are responsible for the hydrophobic interactions between nucleic acids and matrix fibrils. From $\text{in vitro}$ labelling studies the released DNA is more recently synthesised than the bulk material, however the matrix bound RNA appears to label less rapidly than total nuclear RNA.     

The relation of rat liver wet weight to dry weight

To assess a reliable relation between the dry and wet weight of rat livers, the water content of liver samples was determined by freeze drying. The ratio between wet and dry weight of the livers turned out as 3.33 +/- 0.3 for male and 3.28 +/- 0.24 for female rats. Thus for calculations a value of 3.3 can be used irrespective of the sex of rats.     

The relation of rat liver wet weight to dry weight

Summary To assess a reliable relation between the dry and wet weight of rat livers, the water content of liver samples was determined by freeze drying. The ratio between wet and dry weight of the livers turned out as 3.33±0.3 for male and 3.28±0.24 for female rats. Thus for calculations a value of 3.3 can be used irrespective of the sex of rats.Supported by grants from the Deutsche Forschungsgemeinschaft (Sa 127/8-4)     

The staining properties of halo- and alkyl-thiazolylazophenol derivatives in histochemical staining of cadmium

In the continuous research for a more sensitive chelator for cadmium, halothiazolylazodiethyaminophenol and alkylthiazolylazophenol derivatives were synthesized, and their staining sensitivity for cadmium chelates were examined with respect to a substituent group. 2-(4-Methyl- and 4,5-dimethyl-2-thiazolylazo)-5-diethylami-nonphenol showed low staining density in the 005-sections of cadmium, to date they showed the highest staining sensitivity for cadmium among the thiazolylazophenol analogs. The appropriate combination of substituent groups as based on their electron donative strength may be the decisive factor in determining the staining sensitivity of thiazolylazophenol analogs.     

Quantification of histochemical staining by color deconvolution

OBJECTIVE: To develop a flexible method of separation and quantification of immunohistochemical staining by means of color image analysis. STUDY DESIGN: An algorithm was developed to deconvolve the color information acquired with red-green-blue (RGB) cameras and to calculate the contribution of each of the applied stains based on stain-specific RGB absorption. The algorithm was tested using different combinations of diaminobenzidine, hematoxylin and eosin at different staining levels. RESULTS: Quantification of the different stains was not significantly influenced by the combination of multiple stains in a single sample. The color deconvolution algorithm resulted in comparable quantification independent of the stain combinations as long as the histochemical procedures did not influence the amount of stain in the sample due to bleaching because of stain solubility and saturation of staining was prevented. CONCLUSION: This image analysis algorithm provides a robust and flexible method for objective immunohistochemical analysis of samples stained with up to three different stains using a laboratory microscope, standard RGB camera setup and the public domain program NIH Image.     

Phagosome-lysosome interactions related to erythrophagocytosis in Kupffer cells of fetal rat liver

Kupffer cells of fetal rat liver were examined by ultrastructural cytochemical methods to reveal acid phosphatase (AcPase) activity in lysosomes. Elongated cisternae, 940-1150 A in width containing AcPase reaction product, were identified in these cells. These cisternae were sometimes in continuity with phagosomes containing engulfed erythrocytes. Observations suggest that such cisternae may partly encircle these phagosomes. The relationships of these cisternae to GERL (Golgi Endoplasmic Reticulum Lysosomes) is discussed.     

A histochemical study of liver enzymes involved in glycogen metabolism in the X-irradiated rat

Summary The liver enzymes responsible for the breakdown and synthesis of glycogen from glucose have been investigated cytochemically in rats exposed to 1200 rads of x-irradiation. It was found that significant changes occur in their activities and that amylophosphorylase and amylo-1,6-glucosidase (debranching enzyme), both of which are responsible for the conversion of glycogen to glucose, are markedly inhibited by radiation. A significant inhibition of the activity of 1,41,6 transglucosidase (branching enzyme) was also observed. In contrast, the activity of UDPG-glycogen transglucosylase, which is responsible for the in vivo synthesis of 1,4-polysaccharides, was found to be stimulated.