Giant cell granuloma of the lung.

In a 50-year-old man without bronchopulmonary symptoms a round mass lesion close to the hilum of the right lung was detected in a routine chest x-ray and confirmed by computed tomography. Histological examination of two biopsy specimens did not result in a definitive diagnosis. Therefore thoracotomy with enucleation of the focus was performed. The histological picture of the lesion is characterized by connective tissue proliferation, multinucleated giant cells, ossification, localised hemorrhage, deposits of hemosiderin and foci of foam cells. The findings are interpreted as a giant cell granuloma of the lung.

Are Z-Arg-Gly-Phe-Phe-Leu-MNA and Z-Arg-Gly-Phe-Phe-Pro-MNA suitable substrates for the demonstration of cathepsin D activity?

The suitability of Z-Arg-Gly-Phe-Phe-Leu-MNA and Z-Arg-Gly-Phe-Phe-Pro-MNA for the assessment of cathepsin D activity was tested in biochemical and histochemical experiments. Substrates were dissolved in dimethylformamide and used at 0.1-0.5 mM in various buffers over a pH range of 3.5-7.4. Homogenates of various rat organs and isolated purified enzymes [cathepsin D from bovine spleen, dipeptidyl peptidase (DPP) IV from porcine kidney and rat lung] were used as enzyme sources. Pepstatin, di-isopropylfluorophosphate (DFP), p-chloromercuribenzoate, o-phenanthroline and a series of DPP IV inhibitors were used in inhibitor experiments. At pH 3.5 and 5.0, substrates were used in a two-step postcoupling procedure with aminopeptidase M and dipeptidyl peptidase IV as auxiliary enzymes and Fast Blue BB as coupling agent. Results were compared with those obtained with haemoglobin. Above pH 5.0 substrates were used in a one-step postcoupling procedure. Cryostat sections of snap-frozen or cold aldehyde-fixed tissue pieces of various rat organs and biopsies of human jejunal mucosa were used in histochemical experiments. As in biochemical tests a two-step procedure was used in the pH range 3.5-5.0, but Fast Blue B was used in the second step for the simultaneous coupling. Above pH 5.0 a one-step simultaneous azo coupling procedure was used with Fast Blue B as coupling agent. At pH 3.5 the hydrolysis rate of both synthetic substrates was about 100x lower than that of haemoglobin when cathepsin D from bovine spleen was used.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytoplasmic vacuolation of pancreatic beta cells of rats after oral administration of a derivative of isoquinoline.

beta cells in islets of Langerhans were studied in Sprague-Dawley rats dosed by gavage with 0 (control), 75, 150, 250 or 300 mg/kg body wt/day S-H 966 BS [1-(1-oxido-4-thiomorpholino)-3-(1-piperazinyl)], an isoquinoline derivative. All doses caused a significant and dose-dependent increase in serum glucose (diabetes mellitus). At 250 mg/kg, degranulation of beta cells was discovered after 1 day and vacuole formation after 2 days. Ultrastructural alterations compared well with that seen after treatment with cyproheptadine and other structurally related compounds. The vacuolation of beta cells was fully developed following 6 weeks of daily treatment, when a dose-dependent elevation of blood glucose was first observed. The effects were more severe in males than in females. Lesions were reversible within 6 weeks except at 300 mg/kg in males.

The influence of castration on cell proliferation in the adrenal cortex of female rats.

Proliferation kinetics in the zones of the adrenal cortex of female rats after castration are reported. Following ovariectomy a wave of increased cell proliferation was observed in the zona glomerulosa and zona fasciculata after 1 and 2 days respectively. Increased cell proliferation in the zona reticularis was only slight. Cell proliferation returned to the level in control animals at 4 weeks. It is assumed that the regulative process following ovariectomy takes place directly in the adrenal cortex but it is possible that steroid metabolism in the liver is of importance. No relationship between the 3H-TdR labelling index (LI) and the mitotic index (MI) was observed.

Autoradiographic investigation of cell proliferation in the adrenal cortex of castrated female rats under the influence of oestradiol.

Ovariectomy and subsequent treatment with ovarian hormone produces a temporary increase in DNA-synthesizing cells in the zona glomerulosa of the adrenal cortex.

Induction of rat alpha 2-macroglobulin in vivo and in hepatocyte primary cultures: synergistic action of glucocorticoids and a Kupffer cell-derived factor.

Turpentine injection into rats elicits enhanced secretion of acute phase proteins including alpha 2-macroglobulin (alpha 2M). Hypophysectomized rats, however, do not respond in this way unless dexamethasone is given together with turpentine. On the other hand, dexamethasone injection alone did not result in an induction of alpha 2M synthesis. When a medium of Kupffer cell cultures was added to hepatocytes, a dose-dependent stimulation of alpha 2M synthesis of up to 4-fold after 10-12 h was observed. However, the presence of low concentrations (10(-9)M) of dexamethasone was essential for the stimulatory effect. We conclude that the acute phase induction of alpha 2M in hepatocytes requires the synergistic action of glucocorticoids and a non-dialysable factor secreted by Kupffer cells.

Comparative studies on the circadian rhythm of corticosterone, lipid and cholesterol levels in adrenals and blood of rats.

The time-dependent relationship of corticosterone, lipids and cholesterol over a 48-hr period was studied in the adrenals and blood of rats. In addition an attempt was made to determine whether there was a reciprocal dependence among these compounds and also a correlation between corticosterone and cholesterol in the adrenals and blood. Corticosterone and cholesterol exhibit a circadian rhythm in the adrenals and blood. The same is true for lipids in the serum. A reciprocal dependence between the compounds in the adrenals and blood could not be demonstrated. Only the time-dependent processes of the corticosterone content in the adrenals and plasma are well correlated with each other. High levels of these steroids in the adrenals are associated with high levels of these steroids in the plasma and vice versa. An inverse correlation between corticosterone and cholesterol exists in the adrenals and in blood. Maximal levels of corticosterone correspond to minimal levels of cholesterol and vice versa.

Ultrastructural and histochemical aspects of prostatic secretion in dog.

The reaction of canine prostatic epithelium was studied ultrastructurally and histochemically (acid phosphatase, zinc) in mature Beagle-dogs 15 minutes after intravenous application of pilocarpine (1 mg/kg body weight). In the normal prostate we found only merocrine secretion with exocytosis of secretory granules. Exocytosis persisted after stimulation with pilocarpine, but was quantitatively greatly increased. Apocrine or holocrine types of secretion were neither found under resting conditions nor under extreme functional stress of the glandular epithelium. Histochemically a distinct decrease of the activity of acid phosphatase and of zinc in the apical part of the epithelium has developed, which corresponds to the ultrastructurally obvious loss of secretory granules.

Autoradiographic studies of the diurnal rhythm of the cell proliferation in the adrenal cortex of the rat.

With the aid of the autoradiographic technique we examined the question of diurnal or circadian rhythm of the cell proliferation as well as the mitotic rate in the three zones of the adrenal cortex of the rat. To evaluate the diurnal rhythm it is necessary to run experiments within 24-hour intervals. In order to examine observed variations as to their reproducibility we prolonged our investigations over a period of 72 hours. The proliferation activity as well as the mitotic rate in the zona glomerulosa show no diurnal rhythm. This is not so in the zona fasciculata. Here we find a correlated circadian rhythm for both parameters.

Contribution to the phosphate-induced nephropathy in the dog. Comparative light and electron microscopic investigations on the proximal tubule after oral application of K2HPO4, Na2HPO4, KCl and NaCl.

20 male beagle-dogs divided into 4 groups were given equimolar amounts of K2HPO4, Na2HPO4, KCl and NaCl daily by gavage over a period of 9 or 22 weeks. A further collective of 5 dogs served as the control. After terminating the experiment the animals were killed and the kidneys were investigated by light- and electron microscopy. No renal damage developed in dogs treated with KCl or NaCl. In those treated with K2HPO4 or Na2HPO4 nephrocalcinosis with disseminated atrophy of the proximal tubule occurred. The following processes in the proximal tubulus could be analysed and pathogenetically classified: the dedifferentiated tubulus epithelial cells undergo advancing autodigestion by multiplication, enlargement and fusion of autophagic vacuoles and autophagolysosomes. After extrusion of the lysosomal bodies into the tubular lumen and the degradation of the intratubular detritus, residues consisting of completely empty and collapsed basement membrane tubes remain. The significance of these findings for clinical diagnostics in relation to the urine is discussed. It is emphasized that, in the case of proven tubulopathy with a negative protein finding and an absence of epithelial cells and casts in the urine, it can be assumed that the protein and epithelial cells were digested intratubularly by lysosomal enzymes.

[Changes in total beta-glucuronidase activity and DNA content of various tissues and organs of newborn rats after prenatal administration of glucocorticoid]. / Veränderungen von beta-Glucuronidase-Gesamtaktivität und DNS-Gesamtgehalt verschiedener Gewebe und Organe neugeborener Ratten nach pränataler Glucocorticoid-Verabreichung.

Quantitative biochemical analyses of the beta-glucuronidase total activity and the total DNA-content carried out on selected parenchymal and mesenchymal organs of 455 Wistar inbred rats (strain Chbb:THOM (SPF) from 56 litters (untreated or treated with 1 mg betamethasone/kg b.w. s.c. on both the 18th and 19th days of pregnancy, applied to the mother animal) led to the following results: The DNA-content of lung, liver, kidney, skin and rib cartilage declined continuously both in the controls and in the treated group during the postnatal observation period (from the 1st to the 15th day p.p.). The prenatal glucocorticoid administration led to a decline in the DNA-content in comparison to the untreated controls, especially evident in the first postnatal days. The beta-glucuronidase activity of lung, liver and kidney showed an increase in the postnatal development phase to a maximum occurring organ-dependently on different days with a decrease afterwards. Due to glucocorticoid-pretreatment of the mother animals the newborn rats showed significantly higher enzyme values within the first postnatal days than did the untreated controls. Between the 6th and 9th day (depending on the organ) the same beta-glucuronidase activities could be found on pretreated and untreated animals. For the further ages tested in this study (till the 15th day of postnatal development) the enzyme activity of the experimentally treated litters was lower than that of the untreated controls. In connection with related findings, especially regarding morphological changes in the lung, the induction of beta-glucuronidase as well as the decrease in the DNA-content due to prenatal glucocorticoid pretreatment are discussed in terms of a maturation acceleration.

[Animal studies on the tolerability of AR-L 115 BS (author's transl)]. / Tierexperimentelle Untersuchung zur Verträglichkeit von AR-L 115 BS.

Acute and subacute toxicity studies as well as reproduction-toxicologic investigations were carried out on the positive-inotropic substance 2[(2-methoxy-4-methylsulfinyl)phenyl]-1H-imidazo[4,5-b]pyridine (AR-L 115 BS) in various species. No undesirable side-effects or organ damage were observed in baboons after oral application of 2.5, 10 and 30 mg/kg AR-L 115 BS for 13 weeks. Beagle dogs, female miniature pigs and rats had brown-coloured urine after the substance. In miniature pigs and rats this was only observed at high dosage (mini-pig: 10 and 30 mg/kg; rat: 200 mg/kg). Changes in the ECG could be seen in the dose range of 20--40 mg/kg (dog i.v.), 30 mg/kg (dog p.o) and 200 mg/kg (rat p.o). Non-inflammatory mitral valve alterations (i.v. and p.o) as well as spot- and ring-shaped endocardial scleroses underneath the aortic valves in the region of the left cardiac outflow tract (30 mg/kg, p.o.) were found especially in female beagle dogs in the same dose range. The alterations are regarded as being of haemodynamic origin in view of the increased myocardial contractility caused by AR-L 115 BS. Phonocardiographic findings support this view. Besides, several dogs (30 mg/kg) and rats (200 mg/kg) showed myocardial scars. All male miniature pigs which had received 30 mg/kg AR-L 115 BS demonstrated an increase in heart and mitral valve weights. The results of the reproduction-toxicologic investigations gave no indication of an embryotoxic or mutagenic effect for the substance AR-L 115 BS. After comparing the results it can be concluded that the dog is the most sensitive of the species under investigation and that the results obtained in this species should not be generalised. Biochemical tests showed very similar metabolic patterns for AR-L 115 BS in baboons and man. From this it can be deduced that the results obtained in baboons are most likely to be transferable to man.

[Potassium hydrogen phosphate induced nephropathy in the dog. I. Pathogenesis of tubular atrophy (author's transl)]. / Die Kaliumhydrogenphosphat-induzierte Nephropathie des Hundes. I. Pathogenese der Tubulusatrophie.

A nephropathy with severe tubular atrophy was observed in Beagle dogs after oral administration of K2HPO4 for 14 or 38 weeks. We describe the complete lysosomal degradation of atrophying tubular epithelial cells. During two experiments of 14 and 38 weeks duration, respectively, a total of 15 Beagle dogs received 0.8 g K2HPO4/kg body weight daily with their food. All dogs were examined clinically at regular intervals. Renal biopsies were taken in the fourth week from beagles of the 14-week study. Results were compared with those of control dogs. At the end of the experiments the animals were killed and necropsies done. Different stains and histochemical reactions were applied to paraffin sections of the kidneys. Acid phosphatase and beta-glucuronidase were found on cryostat sections. Kidneys fixed by perfusion of five Beagles from the 38-week study and three Beagles of the 14-week study, and from five control dogs, were examined electron microscopically. Ultrahistochemically, acid phosphatase was demonstrated. Clinically, the dogs in both experiments vomited, were cachectic, and had elevated creatinine and blood urea nitrogen. Morphologically, qualitatively identical changes were seen, but the renal damage was most marked at 38 weeks. There were disseminated tubular atrophy (usually of the proximal tubules), focal scar tissue and nephrocalcinosis. The following pathogenesis was established for the lesions of the proximal tubule: Tubular atrophy begins with loss of differentiation of epithelial cells. Enzyme histochemistry, ultrahistochemistry and electron microscopy show an increase in autophagic vacuoles and autophagolysosomes. The lysosomal bodies showing fusion enclose large parts of the cytoplasm as the process continues. Complete lysosomal degradation of epithelial cells and extrusion of large lysosomes into the tubular lumen follow. After complete enzymatic digestion of the intratubular detritus, the residue is empty, convoluted and collapsed tubular basement membrane. Atrophic tubular epithelial cells have many organelle-free zones at their base, which contain fine filamentous material resembling that of the basement membrane. The degradation process described here may explain why clinically the urinary sediment contains few cylinders and epithelial cells and why proteinuria decreases significantly toward the end of the experiment. So far, it is not clear whether the tubular basement membrane is synthesized by the tubular cells, by fibroblasts or by both cell types. The presence of basement membrane-like material in tubular epithelial cells and in parietal epithelial cells of the glomerulus favors the view that epithelial cells produce the basement membranes and that increased production of basement membrane-like material is a sign of loss of differentiation.

[Potassium hydrogen phosphate induced nephropathy in the dog. II. Glomerular alterations (author's transl)]. / Die Kaliumhydrogenphosphat-induzierte Nephropathie des Hundes. II. Glomeruläre Veränderungen.

A disseminated atrophy of the proximal tubule accompanies K2HPO4-induced nephropathy in dogs. These pathologic processes cause glomerular changes that pass through different inflammatory stages and terminate in glomerular sclerosis. Experimental animals, design of the experiment and methods have been described [15]. During the 14-week study we determined the amount of urine (24 hours), protein (mg/dl), protein excretion (mg protein/24 hr) and the macro- and microprotein fraction in the urine by SDS-polyacrylamide gel electrophoresis. Clinical examinations were at 15, 66, and 85 days. Beagle dogs treated with 0.8 g K2HPO4/kg body weight developed significant glomerular selective and unselective protienuria. During the experiment the macroproteins in the urine decreased markedly, and at the last examination (day 85) glomerular proteinuria was no longer detectable by electrophoresis. Morphologically, there were only slight glomerular changes in the biopsy material taken at four weeks. Widespread lesions at 14 and 38 weeks were dilatation of Bowman's space, thickening of the basement membrane, increase in mesangial matrix, interposition of non-argentophilic mesangial matrix into the glomerular basement membrane, protein deposits in the mesangium and parietal basement membrane, formation of crescents, shrinkage of the glomeruli with collapse of glomerular tufts, and finally glomerular sclerosis. The parietal epithelial cells contained cytoplasmic areas that were free of organelles and contained microfilamentous and fine-granular material. These areas were close to the capsular basement membrane. Bundles of filaments within parietal epithelial cells had contact with the basement membrane, thus resembling hemidesmosomes. The sequelae of tubular atrophy are retention of glomerular filtrate and dilatation of Bowman's space, followed by compression and shrinkage of the glomerular tufts, and inflammatory processes within the glomerulus. The latter may be characterized as mesangio-sclerosing, mesangio-proliferative, membrano-proliferative, and extra-capillary glomerulonephritis. The decrease of urinary protein excretion towards the end of the experiment may be related to intratubular lysosomal digestion of cellular and amorphous components.

[Experimental investigations of the content of glycogen in the adrenal gland of the rat after application of dexamethasone, ACTH and a combination of both (author's transl)]. / Experimentelle Untersuchungen über den Glykogengehalt der Rattennebenniere nach Gabe von Dexamethason, ACTH sowie der Kombination Dexamethason/ACTH.

The influence of dexamethasone, ACTH and the combination of both on the glycogen content of the rat adrenal gland was studied. Dexamethasone induces a transient increase of the adrenal glycogen content in a time and dose-dependent manner. The application of ACTH has no effect on the glycogen content. Surprisingly, the increase of the glycogen content induced by dexamethasone is completely blocked by the simultaneous application of ACTH.