The metabolism of shikimate in the rat.

In the rat, shikimate was metabolized and excreted as hippurate, hexahydrohippurate, 3,4,5,6-tetrahydrohippurate, t-3,t-4-dihydroxycyclohexane-r-1-carboxylate and c-3,t-4-dihydroxycyclophexane-r-1-carboxylate, conjugates of catechol and CO2. The metabolism was entirely dependent on various initial microbial transformations in the gut, metabolite formation being suppressed in animals pretreated with antibiotics. Shikimate was not metabolized by mammalian tissues, and products of microbial metabolism were excreted either unchanged or after further biotransformation in the animal tissues.     

The metabolism of cyclohexanecarboxylate in the rat.

In the rat, cyclohexanecarboxylate was metabolized and excreted (mostly in the urine) as hippurate, hexahydrohippurate, 3,4,5,6-tetrahydrohippurate and benzoyl and cyclohexylcarbonyl beta-glucuronides. The pattern of metabolism is dose-dependent. With decreasing dose a progressive increase in the conversion into hippurate occurred. This was largely at the expense of glucuronide formation, although the proportions of hexahydro- and tetrahydro-hippurate were also decreased. The observed formation of hexahydrohippurate and 3,4,5,5-tetrahydrohippurate substantiates the proposed mechanism of aromatization of cyclohexanecarboxylate. It appears that these compounds arise via glycine conjugation of active intermediates in the aromatization process. Hexahydrohippurate and 3,4,5,6-tetrahydrohippurate may occur in the urine of rats as new mitabolities of shikimate, dependent for their formation on microbial metabolism.     

Lymphopoiesis in the rat. II. The effect of thymectomy

Lymphopoiesis with respect to recirculating and non-recirculating small lymphocytes was measured simultaneously in rats thymectomized as adults. Removal of the thymus at four to five weeks of age had a profound inhibitory effect upon the production of recirculating cells, whereas the formation of non-recirculating lymphocytes was only slightly depressed. Thymectomy had approximately the same impact of lymphopoiesis as thymectomy and exposure of the animal to a large dose of whole body X- and γ-irradiation. The latter finding, and the failure of a thoracic duct cell transfusion to augment lymphocyte production, accord with the view that the thymus is the principle intermediate source of recirculating small lymphocytes in the normal, unstimulated animal.     

The cytoarchitecture of the adrenal medulla in the rat.

The disposition of adrenaline and noradrenaline storing cells in the adrenal medulla has been studied in the rat. It has been demonstrated that no morphological or functional reason exists for the follicular designation of clumps of chromaffin cells in the medulla. No significant difference was found between the outer and inner zone as regards the frequency of NA cells by morphometric and statistical methods. The cytoarchitectural differences between adrenaline- and noradrenaline-storing cell arrangement are described. Special features of the interrelation between noradrenaline storing cells, cortical cells and the connective tissue framework are stressed. The light and electron microscopic appearance of two types of cortical cells, assumed to represent different states of functional activity is described.     

The metabolic origin of trigonelline in the rat.

A hypothesis of Mason & Kodicek [(1970) Biochem. J. 120, 515-521] that esterified nicotinic acid in niacytin from cereals is a precursor for trigonelline was investigated in rats. Single oral doses of niacytin resulted in the excretion of trigonelline in urine but only in rats that were niacin-deficient and were fed a cereal diet. These animals were found to have an abnormally permeable intestine, which allowed the uptake of molecules not usually absorbed. Orally administered synthetic [14C]nicotinoyl[3H]methylcellulose was shown to be absorbed by niacin-deficient rats on a cereal diet and [14C]trigonelline was excreted in urine. These data indicate that dietary cereal induces a permeability defect in the intestinal mucosa of niacin-deficient rats, which allows the uptake of macromolecular niacytin. The nicotinoyl pyridine nitrogen atom is then methylated and slow hydrolysis releases trigonelline from the macromolecule.     

The fate of extracellular glutathione in the rat.

When intravenously administered to rats, [U-14C]glycine-labelled GSSG, GSH and its analogue ophthalmic acid were rapidly removed from the blood. In perfusion studies with isolated liver, however, the compounds did not enter the liver tissue. Thus, uptake by this tissue is obviously not responsible for the removal of gamma-glutamyl tripeptides from the blood. Instead, rapid hydrolysis of the tripeptides was observed. The undegraded tripeptides were only detected in the blood immediately after administration. Within tissue the degradation product glycine accounted for all the radioactivity. After intravenous injection of the labelled tripeptides the radioactivity accumulated first in the kidney, as shown by autoradiographic studies and chemical analysis of different tissues. The hydrolysis of the gamma-glutamyl tripeptides decreased markedly after the renal arteries were clamped. These observations strongly suggest that renal tissue is the principal site of the degradation of the tripeptides. Inhibition studies and experiments with isolated renal tubules revealed that gamma-glutamyl transpeptidase catalyses the fast hydrolysis of the extracellular peptides. The results indicate that, when entering the extracellular space, glutathione and its analogues are completely hydrolysed and must be resynthesized after reuptake of the constituent amino acids. It is concluded that the degradation occurs mainly on the luminal surface of the renal brush-border membrane and that gamma-glutamyl transpeptidase is a glutathionase acting on extracellular glutathione.     

The radiosensitivity of rat thymocytes.

gamma-Irradiation in vitro apparently blocked a plasma-membrane associated, superoxide-producing, NADPH oxidase in rat thymocytes. Differential centrifugation of the mixed thymocytes indicated the smaller lymphocytes (approx. 6 microns diameter) to be the radiosensitive population. The oxidase system co-isolated in part with thymus nuclei and could be solubilized by detergent treatment [Bellavite, Jones, Cross, Papini & Rossi (1984) Biochem. J. 223, 639-648]. Endogenous NADPH was the rate-limiting component for superoxide formation in vitro. The level of NADPH was lowered by gamma-irradiation, an effect mimicked by GSSG in the presence of 50 microM-ZnCl2 to inhibit GSSG reductase. These findings are suggested as the metabolic basis for interphase death of small lymphocytes exposed to ionizing radiation.     

Os penis of the rat. V. The distal cartilage process

The morphogenesis and morphology of the distally positioned cartilage of the os penis, the processus cartilagineus, are described in rats aged from 1 to 100 days. Based on observations of metachromacy of the process stained with toluidine blue it was found that a processus cartilagineus only exists in the period between 35 and 50 days of age. Before 35 days, the structure consists of connective tissue proper, and after 50 days the cartilage starts to calcify partially. The present paper also initiates studies of experimentally caused alterations of the normal development of the processus cartilagineus by subjecting 35-day-old rats to castration, with subsequent sacrifice at 100 days. Castration at that age causes a complete interruption of normal development of the processus cartilagineus as the structure in 100-day-old castrated rats has distinct morphological characteristics in common with those of 35-day-old normal rats. The present paper, thus, confirms that normal development of the processus cartilagineus seems to be male-hormone-dependent.     

The regulation of the biosynthesis of ubiquinone in the rat.

The urinary excretion of p-hydroxybenzoate was not altered by ubiquinone feeding, but, although decreased considerably, was not eliminated in protein deficiency. The incorporation of p-hydroxy[U-14C]benzaldehyde into ubiquinone in vivo increased in cold-exposed and p-chlorophenoxyisobutyrate (clofibrate)-fed rats, and these changes were parallel with the changes in the incorporation of [2-14C]mevalonate under these conditions. Starvation, cholesterol feeding and cholic acid feeding resulted in the decreased incorporation of p-hydroxy[U-14C]benzaldehyde into ubiquinone, confirming the decreased ubiquinone synthesis. Feeding exogenous ubiquinone increased the hepatic ubiquinone concentration, but did not cause any decrease in the incorporation of p-hydroxy[U-14C]benzaldehyde into ubiquinone, indicating the absence of a feedback control.     

Os penis of the rat. IV. The proximal growth cartilage

Histomorphological and histochemical aspects of the proximal cartilage of os penis and its surrounding perichondrium in 60 rats aged between 1 and 100 days are described. Comparisons at 11-14 days with the mandibular condylar cartilage reveal a slight difference in their general morphological composition. The developmental changes which take place in the os penis cartilage reveal histomorphologic events, some of which may be brought into agreement with previous observations of patterns of transformations of the bone. Observations on an age-dependent morphological appearance of the area adjacent to the proximal surface of the cartilage suggest certain agreements between the mandibular angular cartilage and the os penis cartilage. The study of phosphomonoesterases in the os penis cartilage and its perichondrium reveals significant, unexplained differences in the distribution of alkaline phosphatase between this cartilage and the mandibular condylar cartilage.     

The number of acetylcholinesterase molecules in the rat megakaryocyte.

A megakaryocyte cell series from rat bone marrow has been examined by the isotopic di-isopropyl fluorophosphate (DFP) method for esterases. After complete reaction with 32P-DFP, the numbers of DFP-reacted molecules inindividual cells havebeen determined by beta trackauto-radiography. Previous work has shown the percentage of organophosphate-sensitive sites in these cells which can be taken as active centers of acetylcholinesterase (AChase). Combining these data, the absolute numbers of organophosphate-sensitive esterase molecules and AChase molecules per cell were determined. Histograms show a narrow spread of values within each of four size classes from megakaryoblast to fully mature megakaryocyte, but, with means increasing 4-fold through this series, approximately in proportion to cell volume. A rat megakaryoblast has 2 X 10(6) AChase molecules, and a megakaryocyte (of 48-micro diameter) has 7.6 X 10(6) molecules. The apparent turnover number of the enzyme for intracellular reaction with substrate is calculated and compared with turnover numbers available for other AChases.     

The biliary excretion of circulating asialoglycoproteins in the rat.

Following intravenous injection into the rat a small proportion (0.5 – 3.0%) of asialo α₁-acid glycoprotein, asialo fetuin, asialo CEA¹ and native CEA are excreted in an apparently unchanged form in the bile. The maximum excretion rate occurs one hour after injection in all cases. The possibility of a novel pathway for glycoprotein uptake by the liver is discussed.     

The distribution of glutathione in the rat liver lobule.

A quantitative cytochemical method was developed for measuring the GSH (reduced glutathione) content of hepatocytes in different regions of the rat liver lobule. Use of this method enabled us to show that GSH is not evenly distributed within the rat liver lobule. The hepatocytes located within 100 micrometer of the central vein contain much less GSH than do those in other regions of the rat liver lobule. We suggest that this partially explains the peculiar susceptibility of these cells to electrophilic attack by toxic metabolites formed via the microsomal cytochrome P-450 system.