The Catabolism of Plasma Albumin in the Rabbit : Its rate and regulation

From I¹³¹-albumin studies and previously defined mathematical formulations, rates of breakdown were estimated for native plasma albumin in rabbits. These rates of catabolism per unit weight of animal were remarkably constant and were independent of variations in the steady state values of albumin concentration in the plasma. These results imply that, at least between animals, the breakdown of plasma albumin follows a kinetic process of approximately zero order. It seems plausible that the process operates similarly in individual animals, and hence that albumin is maintained at normal steady state levels in the healthy animal primarily by means of a regulated rate of synthesis.     

The Role of the Gut in Albumin Catabolism : I. Studies in the jejunoileectomized rabbit

I¹³¹-albumin metabolic studies were carried out in 5 rabbits before, 3 weeks after, and several months after removal of 70 to 90 per cent of the jejunum and ileum. A sixth animal was studied before and 11 weeks after a sham operation. During postoperative experiments, the animals were found to be in a highly unsteady state with large losses of albumin from the vascular compartment. Despite these losses, the plasma albumin concentration was maintained at a relatively constant level. No decrease in the albumin efflux occurred following nearly complete jejuno-ileectomy. The data suggest that albumin catabolism is a first order process.     

The Role of the Gut in Albumin Catabolism : II. Studies in enterectomized rabbits

Using I¹³¹-albumin tracer methods, albumin breakdown rates were estimated in 7 rabbits following extensive gastrointestinal resection, including 5 with nearly complete enterectomy, 1 with total gastroenterectomy, and 1 with gastrectomy only, and in 4 sham operated rabbits. Breakdown rates in the resected animals varied from 48 to 187 per cent of the corresponding controls, with an average of 96 per cent. It is concluded that no more than one-half, and probably much less, of albumin breakdown occurs in the gastrointestinal tract.     

Calculation of the Albumin Catabolic Rate in the Non-Steady State

Methods which are in current use for the calculation of the albumin breakdown rate apply only to the steady state animal. In this paper a simple but more general method based on analyses of I¹³¹-albumin tracer data is presented. It utilizes easily measured plasma specific activity and excretory data and is equally applicable to the steady and non-steady states.     

The Purity of Radioiodide-I131 Assessed by in Vivo and in Vitro Methods

Between 41 and 94% of the radioactivity of 24 preparations of I¹³¹ supplied without cysteine preservative was non-iodide on chromatographic analysis. Extraneous radio-activity was essentially absent from I¹³¹ supplied with cysteine. It was converted to iodide-I¹³¹ by 10^-3 M cysteine or iodide but not by incubation at pH 2. The average thyroid uptake of I¹³¹ containing extraneous radioactivity was significantly lower than the uptake of I¹³¹ free from non-iodide impurity in 16 human subjects measured under controlled conditions and in a random group of 669 patients. Incubation of samples of I¹³¹ containing non-iodide radioactivity with tyrosine and cupric chloride resulted in the non-enzymatic formation of monoiodotyrosine-I¹³¹ either in the presence or absence of thyroid homogenate. Enzymatic formation of monoiodotyrosine-I¹³¹ by thyroid homogenates could be demonstrated only when I¹³¹ free from extraneous activity was used.     

Residualizing and non-residualizing analogues of low-density lipoprotein as iodine-123 radiopharmaceuticals for imaging LDL catabolism

Low-density lipoprotein (LDL) labeled via direct iodination or via the radioiodinated residualizing moiety tyramine-cellobiose (TC) were compared in rabbits as potential ¹²³I radiopharmaceuticals for imaging sites of LDL catabolism. The tissue deposition of ¹³¹I-TC-LDL after 24 h as determined by dissection was in the major catabolic organs (liver, adrenals, spleen), and its plasma clearance was slower in rabbits with dietary hypercholesterolemia than in normals. ¹³¹I-LDL was unsuitable as a metabolic tracer due to redistribution of catabolites and/or loss of the label before protein degradation, which resulted in little accumulation of radioactivity in catabolic organs and high thyroid uptake. The plasma clearance half-time was similar (ca 22 h) for the two compounds in normal rabbits, but was increased to about 36 h for ¹³¹I-TC-LDL and decreased to approximately 9 h for ¹³¹I-LDL in hypercholesterolemic animals. The were similar with dynamic imaging of control and hypercholesterolemic rabbits using ¹²³I-labeled analogues. ¹²³I-TC-LDL rapidly localized in the liver, with low thyroid accumulation of radioactivity. The hepatic uptake of ¹²³I-LDL was about half that of ¹²³I-TC-LDL, and thyroid sequestration of radioactivity was significant for ¹²³I-LDL but not ¹²³I-TC-LDL. These data suggest that whereas the residualizing ¹²³I-TC-LDL has a pharmacokinetic profile representative of lipoprotein metabolism, the biodistribution of the activity from injected ¹²³I-LDL is complicated by processes other than protein degradation. The results are discussed with regard to nuclear medicine applications in evaluating lipoprotein catabolism in man.     

Art und Menge der Ausscheidung 14C-markierter Verbindungen bei Chlorella pyrenoidosa unter dem Einfluss des osmotischen Drucks der Nährlösung

Chlorella pyrenoidosa was labelled by ¹⁴CO₂ and the nature and amount of excreted organic compounds in nutrient media of different osmotic pressure were determined after a 24 h period. The total rate of excretion of organic bound ¹⁴C was about 4 μg ¹⁴C per mg harvested algal dry matter or 1% of the total ¹⁴C content of the algae at the beginning of the excretion period. The main compounds found in the excretions were unidentified substances with a molecular weight higher than 700, amino acids, organic acids and sugars. The osmotic pressure of the nutrient medium did not affect the total amount of the organic excretions. However, the excreted amounts of some specific compounds differed in respect to the osmotic conditions of the nutrient medium.     

Pharmacokinetics, Tissue Distribution and Excretion of Recombinant Human Parathyroid Hormone 1–84 in Animals

To study the plasma pharmacokinetics and accumulation of the recombinant human parathyroid hormone (rhPTH) (1–84) in rhesus monkeys, and the tissue distribution and excretion profiles of ¹²⁵I-rhPTH (1–84) in rats. The concentration of rhPTH (1–84) in plasma samples were determined by an enzyme immunoassay (EIA) method after subcutaneous and intravenous bolus injection. The tissue distribution and urinary, fecal and biliary excretion patterns of ¹²⁵I-rhPTH (1–84) were investigated by trichloroacetic acid (TCA) precipitation method. Following subcutaneous (sc) administration rhPTH (1–84) in rhesus monkeys, rhPTH (1–84) exhibited rapid absorption and elimination and had no accumulated tendency after successive sc administration. Following sc administration ¹²⁵I-rhPTH (1–84) in rats, the TCA-precipitated radioactivity was widely distributed and rapidly diminished in most tissues. Approximately 83.9 and 6.8 % of the total radioactivity was recovered in urine and feces by 72 h postdosing, respectively; whereas 4.1 % excreted into bile up to 24 h postdosing. The pharmacokinetics of rhPTH (1–84) complied with linear kinetics within the examined dose range following a single sc administration had no accumulated tendency following multiple sc administration in rhesus monkeys. The accumulation of ¹²⁵I-rhPTH (1–84) in tissues/organs examined, appeared to be low in rats. The major elimination route was by urinary excretion.     

The excretion of metabolites of cortisol and aldosterone in male patients on haemodialysis treatment

A single intravenous injection of ¹⁴C-cortisol and ³H-aldosterone was given to four male uraemic patients on haemodialysis (HD) treatment. The excretion of radioactivity was measured during two weeks in urine, HD fluid and faeces. In two patients, who were injected just before dialysis, large amounts of radioactivity were eliminated in the HD fluid (38 % and 56 % for ³H, 45 % and 57 % for ¹⁴C) and minor amounts were found in the urine (< 5 %); in the faeces respectively 32 % and 30 % of ³H and 18 % and 26 % of ¹⁴C were excreted. Two patients who were injected immediately after dialysis (and who also had a somewhat better kidney function) excreted larger amounts of radioactivity in the urine (10 % and 24 % for ³H, 13 % and 41 % for ¹⁴C) and in the faeces (44 % and 62 % for ³H, 29 % and 37 % for¹⁴C), while in the HD fluid respectively 18 % and 4 % of ³H and 30 % and 12 % of ¹⁴C was eliminated. The plasma radioactivity just before and just after dialysis showed a very good correlation (r = 0.96 to 0.99, p < 0.001) with the radioactivity eliminated in the first and last hour of HD treatment. Between HD treatments, the radioactivity in plasma did not change or decreased only very little. This finding suggests that metabolites of Cortisol and aldosterone to be excreted in the faeces, are very quickly removed from the circulation.     

Biliary and duodenal drainage for reducing the radiotoxic risk of antineoplastic 131I-hypericin in rat models

Necrosis targeting radiopharmaceutical ¹³¹I-hypericin (¹³¹I-Hyp) has been studied for the therapy of solid malignancies. However, serious side effects may be caused by its unwanted radioactivity after being metabolized by the liver and excreted via bile in the digestive tract. Thus the aim of this study was to investigate two kinds of bile draining for reducing them. Thirty-eight normal rats were intravenously injected with ¹³¹I-Hyp, 24 of which were subjected to the common bile duct (CBD) drainage for gamma counting of collected bile and tissues during 1–6, 7–12, 13–18, and 19–24 h (n = 6 each group), 12 of which were divided into two groups (n = 6 each group) for comparison of the drainage efficiency between CBD catheterization and duodenum intubation by collecting their bile at the first 4 h. Afterwards the 12 rats together with the last two rats which were not drained were scanned via single-photon emission computerized tomography/computed tomography (SPECT/CT) to check the differences. The images showed that almost no intestinal radioactivity can be found in those 12 drained rats while discernible radioactivity in the two undrained rats. The results also indicated that the most of the radioactivity was excreted from the bile within the first 12 h, accounting to 92% within 24 h. The radioactive metabolites in the small and large intestines peaked at 12 h and 18 h, respectively. No differences were found in those two ways of drainages. Thus bile drainage is highly recommended for the patients who were treated by ¹³¹I-Hyp if human being and rats have a similar excretion pattern. This strategy can be clinically achieved by using a nasobiliary or nasoduodenal drainage catheter.     

Extracellular Organic Carbon from Some Green Algae

A preliminary study of the extracellular organic carbon in cultures of Ankistro desmus falcatus, Scenedesmus quadricauda, Chlorella pyrenotdosa and Crucigenia tetrapedia have been performed. Carbon analyses, parallel to the increase in cell numbers, were made with an infrared analyzer after oxidation of the organic carbon to CO₂. During the phase of declining relative growth rate and the stationary phase of growth the species excreted organic carbon. For Scenedesmus and Ankistrodesmus an uptake period took place before the excretion period. Approximately 500 · 10⁶ cells/1 were required until excretion started for the former species. The highest figures for the organic carbon liberated by the four species ranged between 3 and 9 mg/l (approx. 8–23 mg organic matter/I). When distributed per cell the extracellular carbon corresponded to 1–3 μg · 10^?6 with a mean for the average excretion for the species tested of 6 · 10^?8μg/cell · day. For Scenedesmus the extracellular carbon was calculated to be 2–5 per cent of the cell-bound carbon.     

Biotransformation of 3-(3′,5′ -Dichlorophenyl)-5,5-dimethyl oxazolidine-2,4-dione

Using 3-(3′,5′-dichlorophenyl)-5,5-dimethyloxazolidine-2,4-dione labeled with ¹⁴C or ³H, absorption, excretion, and tissue distribution in male Wistar rats were studied, and metabolites excreted were identified. At the dosage rates of 100, 300, 1000 and 3000 mg/kg, the maximum excretion of orally administered radioactivity occurred within 24 hr. Increase in the dosage rate was paralleled by decrease in the proportion of urinary elimination. Essentially all the radioactivity was excreted in 2 weeks. DDOD level was generally low in most tissues. Adipose tissue contained higher radioactivity compared with others. Most of the urinary metabolites identified were characterized by hydroxylation at the 4′ position of the benzene ring moiety, and hydrolytic or oxidative modification of the oxazolidine ring portion.     

Effect of vitamin K on the excretion of cholesterol and degradation products in rats

The effects of vitamin K₁ and K₂ on the fecal excretion of the radioactivity from the rats given cholesterol-4^?14C have been studied.There was a significant increase in the radioactivity excretion by the administration of vitamin K, and 23.3–31.5% of the total radioactivity injected were excreted in a week in the feces of the rats administered vitamin K, while 16.3–17.6% in the control animals.The change was due entirely to the increase in fecal bile acid excretion but no significant change was observed in neutral sterol output. The implications of these findings were discussed.     

Excretion of estrone,estradiol, and progesterone in the urine and feces of the female cotton-top tamarin (Saguinus oedipus oedipus)

The excretion of three gonadal steroids was studied in the urine and feces of female cotton-top tamarins (Saguinus oedipus oedipus). Each steroid, ¹⁴C-estrone, ¹⁴C-estradiol, and ¹⁴C-progesterone, was injected into a separate female cotton-top tamarin. Urine and feces were collected at 8 hr intervals for 5 days on the three tamarins. Samples were analyzed to determine the proportion of free and conjugated steroids. Steroid excretion patterns were determined by sequential ether extraction, enzyme hydrolysis, and chromatography. Labeled estrone was excreted in a slow and continuous manner into the urine (57%) and feces (43%) with 90% of the steroid conjugated. The nonconjugated form had an elution profile identical to ³H estrone, but the conjugated portion was not completely hydrolyzed by enzyme. Labeled estradiol was excreted primarily in the urine (87%) and was released rapidly. Over 90% of the injected ¹⁴C-estradiol was excreted in urine as a conjugate, of which 41% was converted to an estrone conjugate and the remaining 59% was excreted as a polar estradiol conjugate. Labeled progesterone was excreted primarily in the feces (95%), 61% of which was free steroid. Four to six individual peaks of radioactivity were found when using celite chromatography and high performance liquid chromatography (HPLC), indicating that progesterone is metabolized into several urinary and fecal metabolites. One of these peaks matched ³H-progesterone and others may be pregnanediols, pregnanetriols, and 17-hydroxyprogesterone. These steroidal excretion patterns help explain the atypical hormonal patterns seen during the tamarin ovarian cycle.     

ALGAL EXCRETION AND BACTERIAL ASSIMILATION IN HOT SPRING ALGAL MATS1

Benthic algal-bacterial mats are present in the effluents of alkaline hot springs at temperatures between 50 and 73 C. The thin surface layer is composed of the unicellular blue-green alga Synechococcus lividus. Also present in the surface layer and forming thick, orange mats beneath it, are filamentous, phototrophic, gliding bacteria of the genus Chloroflexis, also capable of heterotrophic growth. The very low species diversity and the constancy of the hot spring environment, make these mats a good ecosystem for studying the transfer of nutrients from the algae to the bacteria. To determine whether the alga might supply organic materials to the bacterium, excretion by natural populations of S. lividus was studied in the field by means of short-term radioisotope experiments. Under optimal conditions for photosynthesis, between 3 and 12% of the total ¹⁴C fixed was excreted as ¹⁴C-labeled organic compounds. Variations in cell density at concentrations of S. lividus approximating those found in the mat had no effect on the percentage excretion. However, at cell densities below a threshold, level, the percentage excretion increased with diminishing cell density. Except at very low light intensities the percentage of fixed carbon excreted, was very similar for all light intensities tested. Excretion at temperatures approaching the upper limit for growth was not significantly different from the percentage excretion values observed at lower temperatures. ¹⁴C-labeled organic compounds excreted during algal photosynthesis could be subsequently assimilated by natural populations of the bacteria present in the mat.     

Thiolytic cleavage of the anti-tumour compound 4′-(9-acridinylamino)-methanesulphon-m-anisidine (m-AMSA,NSC 156 303) in blood

4′-(9-acridinylamino)methanesulphon-m-anisidide (m-AMSA), a compound with a broad spectrum of experimental anti-tumour activity, was found to have a short biological half-life in mice bearing L1210 leukaemia. The fate of m-AMSA [³H]-labelled in the acridine nucleus, was determined following injection into mice. There was rapid formation of covalent adducts with plasma proteins. Adducts were also formed in freshly isolated blood samples following incubation at 37°C, and were found to be highly fluorescent. The formation of adducts was accompanied by a decrease in the free thiol concentration in plasma, and the concomitant addition of radioactivity from [³H]acridine nuclei. Acid or alkaline hydrolysis of the plasma protein adduct liberated acridone, while digestion with a protease produced unstable fluorescent compounds. A comparison of the rates of acid hydrolysis of the adducts and of model compounds suggested that the adducts were produced as a result of nucleophilic attack at the C-9 position of m-AMSA by protein thiol groups. The side chain of m-AMSA was liberated as 4-amino-3-methoxymethanesulphonanilide. Several congeners of m-AMSA were shown to form similar or identical adducts both in vivo and in vitro, and at rates which correlated with their reactivity towards simple organic thiols.     

The use of formaldehyde-treated 131-I-albumin in the study of digestive vacuoles and some properties of these particles from mouse liver

The trichloroacetic acid-soluble radioactivity released during incubation of mouse liver particles containing intravenously injected formaldehyde-treated ¹³¹I-albumin consisted almost entirely of ¹³¹I-iodotyrosine. The material was shown to be excreted into the medium and was not due to disruption of the particles by acid. Triton X-100 or the absence of sucrose in the medium inhibited hydrolysis of the particle-associated labeled protein. This inhibition was due to disruption of the digestive vacuoles and dilution of the protein and cathepsins in the suspending medium. These results and other experimental evidence strongly suggest that the ¹³¹I-albumin-containing liver particles are digestive vacuoles. The results also establish that ¹³¹I-albumin may be used to study these vacuoles. High concentrations of sucrose (1 M) inhibited degradation of intraparticulate protein. However, 1 M salts inhibited only the rate of the digestion. Sucrose had an inhibitory effect on a crude cathepsin preparation, and salts stimulated the activity when ¹³¹I-albumin was used as substrate. The effect of high sucrose concentrations as an inhibitor of protein hydrolysis within digestive vacuoles was, therefore, most likely due principally to an inhibition of cathepsin activity within the vacuoles. The effect of salt was probably caused by a stimulation of both intra- and extra-particulate cathepsin activities, although 0.5–1.0 M KCl appeared to protect the particles.     

Analysis of elimination mechanisms of some 99mTc-complexes

The biological behaviour of complexes of ^99mTc with aminopolycarboxylic and aminocarbohydroxamic ligands EDTA (ethylenediaminetetraacetic acid), DTPA (diethylenetriaminepentaacetic acid), EDTAH (ethylenediaminetetraacetohydroxamic acid) and HIDAmH (N-2-hydroxyethyl-N-carboxymethyl-aminoacetohydroxamic acid) was studied in rabbits. The pharmacokinetic parameters determined in intact rabbits were compared with the results obtained in the study of renal and hepatic clearance of the complexes under study. Hepatobiliary excretion, which in [^99mTc]EDTA forms 20–30% of the total excreted amount, is of negligible magnitude in the other ^99mTc-complexes studied (<2%). Their renal clearance is not influenced by the inhibition of tubular secretion with probenecid. Binding to plasma proteins increases in the order [^99mTc]DTPA < [^99mTc]EDTA <[^99mTc]HIDAmH <[^99mTc]EDTAH and the elimination half-life increases in the same order. The value of renal clearance of the complexes studied related to inulin clearance correlates well with the fraction of the free drug in the plasma. In rabbits the complexes under study are excreted mainly by the mechanism of glomerular filtration in the kidney.     

ORGANIC COMPOUNDS RELEASED FROM A NATURAL POPULATION OF EPIBENTHIC BLUEGREEN ALGAE1

Extracellular release of dissolved organic compounds by the bluegreen algal community of a brackish marsh was studied using ¹⁴C techniques. Mannitol and trehalose were identified as the most commonly released compounds. The proportions of these two extracellular compounds varied in response to light intensity and the water potential of the environment. The presence of mannitol, in particular, suggests that excretion of organic compounds in natural situations is a function of osmotic adjustment.     

TRANSFER OF N2 AND CO2 FIXATION PRODUCTS FROM ANABAENA OSCILLARIOIDES TO ASSOCIATED BACTERIA DURING INORGANIC CARBON SUFFICIENCY AND DEFICIENCY1

Transfer of N₂ and CO₂ fixation products from the bloom forming blue-green alga, Anabaena oscillarioides Bory, to attached and free swimming bacteria is common during active growth of the former. Incubation with ¹⁵N₂ and ¹⁴CO₂ followed by size fractionation filtration reveals that: i) magnitudes of fixed N and C excretion, relative to N₂ and CO₂ fixation, are dictated by dissolved inorganic carbon (DIC) availability for A. oscillarioides photosynthetic production, ii) associated bacteria exhibit preferences for recently fixed excreted N compounds, iii) bacterial utilization of excreted N is independent of ambient light conditions, and iv) lag times between N₂ fixation and detectable bacterial assimilation of excreted fixed N compounds are ca. 1–2 h. Both ¹⁴NH₄Cl dilution and Hg(NH₃)₂ Cl₂ precipitation techniques indicate that NH₃ is a major excretion product from A. oscillarioides, particularly during DIC limited growth. Active N and C excretion and transfer to associated bacteria are features of viable A. oscillarioides filaments. Hence, transfer of these metabolites reflects complex mutualistic, and possibly symbiotic associations rather than solely signaling senescence.