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Study design:Experimental, controlled study.Objective:To examine the effects of whole-body vibration (WBV) on bone mass and trabecular bone microstructure (TBMS) during the early stage in juvenile rats with spinal cord injury (SCI).Setting:Studied at the Kio University in Japan.Methods:Thirty-four 8-week-old male Wistar rats were divided into 3 groups: the SCI group, the sham-operation group (SHAM) and the SCI+WBV group. WBV started on the 8th day after SCI. After 1 or 2 weeks of WBV treatment, measurements of tissue mineral density, trabecular bone mineral content (BMC) and parameters of TBMS were obtained by scanning the proximal tibias with x-ray micro-computed tomography. Serum levels of osteocalcin (OC) and of tartrate-resistant acid phosphatase 5b (TRACP 5b) were measured with ELISA.Results:BMC, volume bone mineral density, bone volume (BV), BV fraction (BV/tissue volume) and connectivity density (Conn.D) of TBMS parameters were significantly higher in SCI+WBV rats than in SCI rats after 2-week WBV. The BMC and BV/TV of bone mass index correlated well with Conn.D, suggesting the preservation of Conn.D. induced by WBV. SCI+WBV rats showed a decrease in serum OC after 1-week WBV, but a quick recovery from that after 2-week WBV. There was no difference in serum TRACP 5b among the 3 groups throughout the experimental period.Conclusion:WBV treatment could attenuate the bone deterioration that occurs during the early stage in juvenile rats with SCI. In a clinic, this early WBV intervention may be an effective rehabilitation modality for preventing bone fragility in SCI patients.

Background: Protein C inhibitor (PCI) plays a role in multiple biological processes including fertilization, coagulation, fibrinolysis and kinin systems. Objectives: We hypothesized that PCI participates in the pathogenesis of pulmonary hypertension. To demonstrate this, we compared the development of pulmonary hypertension in mice overexpressing PCI in the lung with wild‐type (WT) mice. Pulmonary hypertension was induced by s.c. injection of 600 mg kg−1of monocrotaline weekly for 8 weeks. Results: Right ventricular arterial pressure was significantly increased in monocrotaline‐treated WT mice compared with that in monocrotaline‐treated transgenic mice. Bronchoalveolar lavage fluid (BALF) levels of thrombin–antithrombin complex, monocyte chemoattractant protein‐1 and platelet‐derived growth factor, and the plasma level of tumor necrosis factor‐αwere significantly increased in monocrotaline‐treated WT mice as compared with monocrotaline‐treated PCI transgenic mice. Increased level of PCI‐thrombin complex was detected in BALF from monocrotaline‐treated PCI transgenic mice as compared with saline‐treated PCI transgenic mice. Conclusions: This study showed that increased expression of PCI in the lung is protective against monocrotaline‐induced pulmonary hypertension, suggesting a potential beneficial effect of PCI for the therapy of this disease.

The effect of 24,25-dihydroxyvitamin D3 [24,25(OH)2D3] on 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] metabolism was examined in rats fed on a low-calcium diet. These rats exhibit hypocalcaemia, high urinary cyclic AMP excretion, a markedly elevated serum 1,25(OH)2D concentration and low serum concentrations of both 24,25(OH)2D and 25(OH)D. When the rats are treated orally with 1, 5 or 10 micrograms of 24,25(OH)2D3/100 g every day, there is a dramatic decrease in serum 1,25(OH)2D concentration in a dose-dependent manner concomitant with an increase in serum 24,25(OH)2D concentration. Serum calcium concentration and urinary cyclic AMP excretion are not significantly affected by the 24,25(OH)2D3 treatment, which suggests that parathyroid function is not affected by the 24,25(OH)2D3 treatment. The 25(OH)D3 1 alpha-hydroxylase activity measured in kidney homogenates is markedly elevated in rats on a low-calcium diet but is not affected by any doses of 24,25(OH)2D3. In contrast, recovery of intravenously injected [3H]1,25(OH)2D3 in the serum is decreased in 24,25(OH)2D3-treated rats. Furthermore, when [3H]1,25(OH)2D3 is incubated in vitro with kidney or intestinal homogenates of 24,25(OH)2D3-treated rats there is a decrease in the recovery of radioactivity in the total lipid extract as well as in the 1,25(OH)2D3 fraction along with an increase in the recovery of radioactivity in the water-soluble phase. These results are consistent with the possibility that 24,25(OH)2D3 has an effect on 1,25(OH)2D3 metabolism, namely that of enhancing the degradation of 1,25(OH)2D3. However, because a considerable proportion of the injected 24,25(OH)2D3 is expected to be converted into 1,24,25(OH)3D3 by renal 1 alpha-hydroxylase in 24,25(OH)2D3-treated rats, at least a part of the decrease in serum 1,25(OH)2D concentration may be due to a competitive inhibition by 24,25(OH)2D3 of the synthesis of 1,25(OH)2D3 from 25(OH)D3. Thus the physiological importance of the role of 24,25(OH)2D3 in regulating the serum 1,25(OH)2D concentration as well as the mechanism and metabolic pathway of degradation of 1,25(OH)2D3 remain to be clarified.

25-Hydroxy-24-oxovitamin D3 (25(OH)24-oxo-D3), a metabolite of 25-hydroxyvitamin D3, has been chemically synthesized. The ultraviolet, mass, infrared, and proton nuclear magnetic resonance spectra of the 25(OH)24-oxo-D3 were identical with those of the natural product isolated from chick kidney incubates. The oxo compound showed biological activity similar to 24,25-dihydroxyvitamin D3 (24,25(OH)2D3) in vitamin D-deficient chicks in enhancing intestinal calcium transport and bone calcium mobilization activities. Although 25(OH)24-oxo-D3 partially restored the impaired eggshell weights of Japanese quails fed a vitamin D-deficient diet, it was much less potent than 25-hydroxyvitamin D3 or 1 alpha, 25-dihydroxyvitamin D3. In addition, there was no effect on the calcification of medullary bone. When 25(OH)24-oxo[3H]D3 was incubated with kidney homogenates from vitamin D-deficient chicks, it was metabolized to [3H]1 alpha, 24,25-trihydroxyvitamin D3 and a metabolite which was eluted in a region between authentic 24R,25(OH)2D3 and 1 alpha, 25-dihydroxyvitamin D3 on high pressure liquid chromatography. In the incubates of kidney homogenates from vitamin D-supplemented chicks, those metabolites were not detected. In vitamin D-supplemented chicks, the recovery of radioactivity in the chloroform phase extracted by the method of Bligh and Dyer was only 50%, while that in vitamin D-deficient chicks was 87%. Moreover, the radioactivity eluted in the 25(OH)24-oxo-D3 fraction from vitamin D-supplemented chicks was only one-fifth of that from vitamin D-deficient birds. The present results indicate that the 24-oxidation of 24,25(OH)2D3 may be a route of inactivation of vitamin D3.

Human promyelocytic leukaemia cells (HL-60) can be induced to differentiate into mature granulocytes in vitro by 1 alpha,25-dihydroxycholecalciferol [1 alpha,25(OH)2D3], the active form of cholecalciferol. The differentiation-associated properties, such as phagocytosis and C3 rosette formation, were induced by as little as 0.12 nM-1 alpha,25(OH)2D3, and, at 12 nM, about half of the cells exhibited differentiation on day 3 of incubation. Concomitantly the viable cell number was decreased to less than half of the control. Among various derivatives of cholecalciferol examined, 1 alpha,25(OH)2D3 and 1 alpha,24R-dihydroxycholecalciferol were the most potent in inducing differentiation, followed successively by 1 alpha,24S-dihydroxycholecalciferol, 1 alpha-hydroxycholecalciferol, 25-hydroxycholecalciferol and 24R,25-dihydroxycholecalciferol. A cytosol protein specifically bound to 1 alpha,25 (OH)2D3 was found in HL-60 cells. Its physical properties closely resembled those found in such target tissues as intestine and parathyroid glands. 1 alpha,25(OH)2D3 bound to the cytosol receptor was transferred quantitatively to the chromatin fraction. The specificity of various derivatives of cholecalciferol in inducing differentiation was well correlated with that of their association with the cytosol receptor. These results are compatible with the hypothesis that the active form of cholecalciferol induces differentiation of human myeloid leukaemia cells by a mechanism similar to that proposed for the classical concept of steroid hormone action.

The effect of cholecalciferol and its metabolites on ornithine decarboxylase activity was investigated in the duodenal mucosa of vitamin D-deficient chicks. The duodenal ornithine decarboxylase activity decreased in animals fed a vitamin D-deficient diet and its retarded activity was increased dose-dependently by a single injection of cholecalciferol. Among various metabolites of cholecalciferol tested, 1 alpha, 25-dihydroxycholecalciferol [ 1 alpha, 25 (OH)2D3] was the most potent stimulator. Stimulation of the enzyme activity was detected as early as 2h after intravenous administration of 1 alpha, 25 (OH)2D3 and a maximal value was attained at 6 h. The maximal value was 27 times higher than the control. In addition, treatment with 1 alpha 25 (OH)2D3 affected the duodenal content of polyamines. The content of putrescine increased to a value of three times that of the control 6 h after the hormone administration. The spermidine content did not change appreciably. The enhancement of duodenal ornithine decarboxylase activity by 1 alpha, 25 (OH)2D3 occurred in parallel with the enhancement of calcium absorption, which was first detected 3 h after the hormone administration. The enhancement appeared to be tissue-specific. It was observed in every intestinal segment, but was highest in the duodenum. Enzyme activity in other tissues was not influenced appreciably by 1 alpha, 25 (OH)2D3. These results clearly indicate that the duodenal biosynthesis of polyamines is regulated by 1 alpha, 25 (OH)2D3, suggesting the possibility that duodenal ornithine decarboxylase may be involved in the calcium absorption mechanism.

Using four cultured cell models representing liver, keratinocyte, and osteoblast, we have demonstrated that the vitamin D analog, 22-oxacalcitriol is degraded into a variety of hydroxylated and side chain truncated metabolites. Four of these metabolic products have been rigorously identified by high pressure liquid chromatography, diode array spectrophotometry, and gas chromatography-mass spectrometry analysis as 24-hydroxylated and 26-hydroxylated derivatives as well as the cleaved molecules, hexanor-1alpha,20-dihydroxyvitamin D3 and hexanor-20-oxo-1alpha-hydroxyvitamin D3. Comparison with chemically synthesized standards has revealed the stereochemistry of the biological products. Although differences exist in the amounts of products formed with the different cell types, it is apparent that 22-oxacalcitriol is subject to metabolism by both vitamin D-inducible and noninducible enzymes. Time course studies suggest that the truncated 20-alcohol is derived from a side chain hydroxylated molecule via a hemiacetal intermediate and the 20-oxo derivative is likely formed from the 20-alcohol. Biological activity measurements of the metabolites identified in our studies are consistent with the view that these are catabolites and that the biological activity of 22-oxacalcitriol is due to the parent compound. These results are also consistent with recent findings of others that the biliary excretory form of 22-oxacalcitriol is a glucuronide ester of the truncated 20-alcohol.

We examined whether 1α-hydroxyvitamin D 3 (1α(OH)D 3 ) is metabolized into 1α,25-dihydroxyvitamin D 3 (1α,25(OH) 2 D 3 ) in bone. Northern blot analysis indicated that the expression of vitamin D 3 25-hydroxylase mRNA was highest in the liver, followed by the duodenum, calvaria, lung, kidney, skin and long bone, and lowest in the spleen. Of the bone cell fractions isolated from fetal mouse calvaria by a sequential enzymatic digestion, fraction 3, which consisted of mostly osteoblastic cells, showed the highest expression of vitamin D 3 25-hydroxylase mRNA. When either cultured bone cells of fraction 3 or mouse calvaria were incubated with [ 3 H]-1α(OH)D 3 , a radioactive peak which comigrated at the same position as authentic 1α,25(OH) 2 D 3 was found on an HPLC chromatogram. The radioactive fraction obtained from the conditioned media of fetal mouse calvaria was tentatively identified as 1α,25(OH) 2 D 3 by cochromatography with authentic 1α,25(OH) 2 D 3 on three different HPLC systems and a thermal isomerization analysis. These results indicate that 1α,(OH)D 3 is hydroxylated at the 25-position in bones, resulting in the local synthesis of 1α,25(OH) 2 D 3 from 1α(OH)D 3 in the skeletal tissues.

1 alpha,25-Dihydroxyvitamin D3 [1 alpha,25-(OH)2D3] induces fusion of murine alveolar macrophages. This effect was observed in growth medium containing 5% human serum but not in the medium with 5% fetal bovine serum. Unlike 1 alpha,25-(OH)2D3, bacterial lipopolysaccharides (LPS) did not induce fusion of alveolar macrophages. However, both 1 alpha,25-(OH)2D3 and LPS activated alveolar macrophages, as measured by glucose consumption, increase in Fc receptors, and induction of cytotoxicity. The number of Fc receptors on the surface of multinucleated giant cells induced by 1 alpha,25-(OH)2D3 was much smaller than that on the surface of mononuclear macrophages treated with the hormone. These results indicate that 1 alpha,25-(OH)2D3 induces both fusion and activation of alveolar macrophages, whereas LPS elicits activation only.

Abstract: The behavior of aluminum compounds in animals was made clear by using non-destructive neutron activation analysis instead of radiotracer method. Basic aluminum sucrose sulfate (BASS) and dried aluminum hydroxide gel (DAHG) were orally administered to rats. The residual BASS and DAHG on gastric walls of rats were determined by neutron activation analysis. It was obtained the following results that BASS was found at the longer time in the stomach. For reason, it is seemed that BASS is hydrolyzed in gastric juice. And it could not find the aluminum in liver, kidney and spleen.

We performed dosing experiments to evaluate the bone mass increasing action of a novel, synthetic vitamin D derivative, 2β-(3-hydroxypropoxy)-1α, 25(OH)2D3 (ED-71), in normal and estrogen-deficient rats. The first experiment consisted of 31 Sprague-Dawley rats, 28 weeks of age. The second experiment consisted of 44 animals who were ovariectomized (OVX) or sham operated at the age of 12 weeks. ED-71 was given twice a week for the duration of 12 weeks. At the end of the experiments, serum chemistries were examined and lumbar vertebrae were assessed histomorphometrically. Serum alkaline-phosphatase levels tended to decrease by ED-71 administration in the first experiment and their elevated values after ovariectomy were also depressed by ED-71 in the second experiment. Serum osteocalcin levels, however, increased by the agent. In the first experiment, cancellous bone volume (BV/TV) increased dose dependently. Bone formation rates (BFR/BS) also increased. In the second experiment, BV/TV significantly decreased by ovariectomy and it increased in ED-71-treated groups, but not in 1α-(OH)D3-treated group. BFR/BS increased by ED-71. Activation frequency did not decreased by ED-71 in either experiment. These data clearly demonstrated that ED-71 administration was capable of increasing the bone mass by stimulating bone formation in normal and estrogen-deficient rats.

Peroxisome proliferators (e.g. clofibric acid) and thyroid hormone play an important role in the metabolism of lipids. These effectors display their action through their own nuclear receptors, peroxisome proliferator-activated receptor (PPAR) and thyroid hormone receptor (TR). PPAR and TR are ligand-dependent, DNA binding, trans-acting transcriptional factors belonging to the erbA-related nuclear receptor superfamily. The present study focused on the convergence of the effectors on the peroxisome proliferator response element (PPRE). Transcriptional activation induced by PPAR through a PPRE was significantly suppressed by cotransfection of TR in transient transfection assays. The inhibition, however, was not affected by adding 3,5,3'-triiodo-L-thyronine (T3). Furthermore, the inhibition was not observed in cells cotransfected with retinoic acid receptor or vitamin D3 receptor. The inhibitory action by TR was lost by introducing a mutation in the DNA binding domain of TR, indicating that competition for DNA binding is involved in the molecular basis of this functional interaction. Gel shift assays revealed that TRs, expressed in insect cells, specifically bound to the 32P-labeled PPRE as heterodimers with the retinoid X receptor (RXR). Both PPAR and TR bind to PPRE, although only PPAR mediates transcriptional activation via PPRE. TR.RXR heterodimers are potential competitors with PPAR.RXR for binding to PPREs. It is concluded that PPAR-mediated gene expression is negatively controlled by TR at the level of PPAR binding to PPRE. We report here the novel action of thyroid hormone receptor in controlling gene expression through PPREs.

Changes in the amount of cytosolic 3,5,3′-tri-iodo-l-thyronine (T3)-binding protein (CTBP) and its activator during administration of l-thyroxine (T4) to thyroidectomized rats were investigated. Thyroidectomy decreased the amount of CTBP in the kidney, whereas the activator was not significantly modified by thyroidectomy. The activator was increased by administration of T4to thyroidectomized rats. The amount of CTBP was also increased by administration of T4. The activator increased the maximal binding capacity (MBC) without changes in the affinity constant for T3binding in CTBP. A T4-induced increase in MBC in cytosol inhibited nuclear T3binding in vitroby competition of T3binding between CTBP and the nuclear receptor.These results suggest that thyroid hormone increases the capacity for cytosolic T3binding through increasing the amount of CTBP and its activator, and that these increases play a role in regulating the amount of T3that binds to its nuclear receptor.Journal of Endocrinology(1989) 123,99–104

The present study was undertaken to clarify the receptor distribution and the pharmacokinetics of 3H-1α(OH)D3, and 3H-1α,25(OH)2D3for comparison. Receptor autoradiography was used after intravenous injection to 3-day-old neonatal rats and radioassay-HPLC after oral application to young adult rats. Corresponding results were obtained from both receptor autoradiography and radioassay. After 3H-1α(OH)D3administration, uptake was delayed but sustained over a long period of time and the concentration of silver grains (autoradiography) or recovered 3H-1α,25(OH)2D3(radioassay) peaked at a lower level. After 3H-1α,25(OH)2D3administration, osteoblast nuclear, whole bone uptake and retention of radiolabeled compound were relatively rapid and short in duration. Nuclear uptake in osteoblasts after administration of 3H-1α(OH)D3was abolished in competition studies with 10-fold unlabeled 1α,25(OH)2D3. These results indicate that 1α(OH)D3continuously supplies osteoblasts with converted 1α,25(OH)2D3and would not spread to the cells because of the low binding affinity of the receptor. Accordingly, 1α(OH)D3appears to have some therapeutic properties different from 1α,25(OH)2D3because of a relatively slow and sustained accumulation of the receptor and less Cmax(pharmacokinetics) compared with 1α,25(OH)2D3.

Cytosolic NADPH-dependent 3,5,3′-triiodo-L-thyronine (T3)-binding protein (CTBP) purified from rat kidney was further characterized in its T3binding and its interaction with nuclei. Pretreatment of the CTBP with NADP induced dithiothreitol (DTT)-dependent T3binding. The DTT-dependent T3binding was increased by NADP in a concentration-dependent manner, and the maximal binding was obtained by 0.1 microM NADP. Higher concentrations of NADP (more than 0.1 µM), however, reduced T3binding. NAD also induced DTT-dependent T3binding, but was very low compared to that induced by NADP. NADPH and NADH did not produce DTT-dependent T3binding. This NADP-activated, DTT-dependent T3binding was characterized as follows: Kafor T3binding was 1.8 × 109M−1, and the maximal binding capacity was 15,000 pmol/mg of protein in the CTBP activated by 0.1 µMNADP. The molecular weight of the CTBP was 58,000 (4.7 S).

The NADPH-dependent cytosolic 3,5,3′-triiodo-L-thyronine(T3)-binding protein (CTBP) has been purified over 30,000-fold from rat kidney by using charcoal extraction, Mono Q-Sepharose, Blue Sepharose CL-6B, and Sephacryl S-200 column chromatography. Purified CTBP had a sedimentation coefficient of 4.7 S, Stokes radius of 32.5Å, and calculated molecular weight of 58,000. The apparently homogeneous protein consisted of a single polypeptide chain with Mrof 58,000 as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Scatchard analysis of T3binding showed that NADPH increases maximal binding capacity without changes in the affinity constant (Ka= 2.43 × 109M−1). Double reciprocal analysis of NADPH and binding capacity gave maximal binding capacity of 16,400 pmol/mg of CTBP, Mr= 58,000. The order of affinity of iodothyronine analogues to purified CTBP was as follows: L-T3= D-T3> triiodothyroacetic acid >L-thyroxine. [125I]T3bound to purified CTBP spontaneously dissociated from CTBP at 20 °C (t½= 22 min) in the absence of NADPH, whereas the dissociation was not observed in the presence of NADPH. The optimal pH for T3binding was 7.2–7.5 Na+, K+, Ca2+, and Mg2+(0-200 mM) did not influence T3binding to CTBP. The purified CTBP did not bind to DNA and was not adsorbed to concanavalin A-Sepharose.

Extensive fusion was induced in mouse alveolar macrophages by treatment with conditioned media obtained from spleen cell cultures treated with 15 micrograms of phytohemagglutinin or concanavalin A per ml or with 12 nM 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3]. The fusion rate was 80-90% on day 3. In addition, 1 alpha,25(OH)2D3 added directly to alveolar macrophages induced fusion of about 35% of the cells on day 3, whereas direct addition of phytohemagglutinin and concanavalin A did not enhance fusion at all. When conditioned media from spleen cell or T cell cultures treated with 12 nM 1 alpha,25(OH)2D3 were applied to a Sephadex G-100 column, a fusion factor (Mr 37,000-70,000) could be separated from 1 alpha,25(OH)2D3. 1 alpha,25(OH)2D3 induced fusion at 0.012-120 nM in a dose-dependent manner both by direct action and by spleen cell-mediated indirect action, but the fusion rate was always much greater in the latter than in the former at each concentration of the vitamin. Of the vitamin D3 derivatives tested, 1 alpha,25(OH)2D3 was the most potent, followed successively by 1 alpha,24R,25-trihydroxyvitamin D3, 1 alpha-hydroxyvitamin D3, 25-hydroxyvitamin D3, and 24R,25-dihydroxyvitamin D3. These results clearly indicate that 1 alpha,25(OH)2D3 induces fusion of mouse alveolar macrophages by both a direct and an indirect mechanism, the latter mediated by spleen cells, probably by T cells.

Syngeneic SL mice inoculated with murine myeloid leukemia cells (M1) all died of leukemia within 30 days. Treatment three times a week with 12.5-50 pmol per mouse of either 1 alpha,25-dihydroxyvitamin D3 [1 alpha,25(OH)2D3], the active form of vitamin D3, or its synthetic analog, 1 alpha-hydroxyvitamin D3 [1 alpha(OH)D3], considerably prolonged the survival time of mice inoculated with M1 cells. 1 alpha(OH)D3 was more effective than 1 alpha,25(OH)2D3 in increasing the survival time of the mice. 1 alpha(OH)D3 also increased the survival time of nude mice inoculated with M1 cells. The 1 alpha(OH)[3H]D3 administered intraperitoneally to tumor-bearing mice was converted very rapidly to 1 alpha,25(OH)2-[3H]D3. The chronic administration of 25 pmol of 1 alpha(OH)D3 to tumor-bearing mice for 30 days caused no appreciable hypercalcemia. These results indicate clearly that 1 alpha,25(OH)2D3 is effective not only in inducing differentiation of M1 cells in vitro, as previously reported [Abe, E., Miyaura, C., Sakagami, H., Takeda, M., Konno, K., Yamazaki, T., Yoshiki, S. & Suda, T. (1981) Proc. Natl. Acad. Sci. USA 78, 4990-4994], but also in prolonging the survival time of mice inoculated with M1 cells.

In order to study whether peripheral action of thyroid hormones is altered in insulin deficiency and to elucidate the biological consequences of alteration of the cytosolic 3,5,3′-tri-iodo-l-thyronine (T3) binding protein (CTBP), we measured malic enzyme, T3-responsive nuclear nprotein, CTBP and nuclear thyroid hormone receptor in the liver and kidney of streptozotocin (STZ)-induced diabetic rats that were treated with or without insulin and/or a receptor-saturating dose of T3. The following results were obtained. 1. Induction of malic enzyme by T3was apparently diminished in diabetic rats. However, supplementary injection of insulin enabled previously given T3to take effect in diabetic rats. 2. T3-responsiveness of other hepatic proteins (nprotein and CTBP) was not altered by insulin in diabetic rats. 3. The level of nprotein was increased by insulin in diabetic rats in vivoand in perfused rat liver, indicating that the hepatic nprotein is a novel insulin-responsive protein. T3and insulin increased the level of nprotein non-synergistically in diabetic rat liver. 4. Hepatic nuclear receptor levels were not altered in diabetic rats. 5. Hepatic CTBP levels were decreased in diabetic rats. This was not due to the toxic effect of STZ. Low CTBP level was only partially increased by insulin after 30 days of diabetic period. Renal CTBP levels were not altered in diabetic rats with or without insulin treatment. These results indicate that reduction of CTBP did not influence the hepatic response to a receptor-saturating dose of T3, although CTBP may regulate the nuclear T3transport, and that fundamental action of a receptor-saturating dose of T3was not attenuated in diabetic rat liver.Journal of Endocrinology(1994) 143, 55–63

Human thyroid hormone receptor (c-erb A protein) produced by Escherichia coliexpression vector plasmid was purified sequentially using polyethylenimine precipitation of DNA, hydroxylapatite column chromatography, ammonium sulphate precipitation, Sephacryl S-300 gel filtration and mono Q-Sepharose column chromatography. These column procedures resulted in 41.3-fold purification of 3,5,3′-tri-iodo-l-thyronine (T3) binding activity over the initial E. coliextract. Purified protein as well as crude preparation showed high-affinity binding to T3. The c-erb A protein enriched by column purification was further purified by electroelution after electrophoresis. Rabbit antibody against the c-erb A protein was prepared and used for the Western blotting analysis. The antibody recognized c-erb A protein but not the bacterial proteins in crude E. coliextract. When partially purified rat hepatic nuclear thyroid hormone receptor was analysed, a 56kDa receptor was specifically recognized by the antibody.

The localization of synapsin I in the rat adrenal medulla was studied using the light- and electronmicroscopic immunohistochemistry. By light microscopy, many dot-like reaction products for synapsin I were recognized to be distributed throughout the medullary tissue. The immunoelectron microscopy clearly revealed that gold particles for synapsin I accumulated in abundance in the nerve terminals forming synapses with the chromaffin cell, while the particles were not localized in the chromaffin cells at all. In the nerve terminal, the gold particles were localized exclusively in the region occupied by synaptic vesicles except for the region just beneath the presynaptic plasma membrane. The synaptic vesicles were frequently linked with the adjacent ones by filamentous structures implicated in synapsin I. It is concluded morphologically that synapsin I is a highly-specific protein for the genuine neuron, and is not detected even in the chromaffin cell which originates from the neural crest.

The effects of vitamin D metabolites on alkaline phosphatase [ALPase; orthophosphoric-monoester phosphohydrolase (alkaline optimum), EC 3.1.3.1] activity, a marker of terminal differentiation, in chondrocyte cultures and growth plates in vivo were examined. In cultures of pelleted rabbit growth-plate chondrocytes, 1,25-dihydroxycholecalciferol (1,25-dihydroxyvitamin D3) increased the contents of DNA and macromolecules containing uronic acid (proteoglycans). It also decreased ALPase activity with an ED50 of less than 1 nM. Other vitamin D3 metabolites, such as 24,25-dihydroxycholecalciferol and 25-hydroxycholecalciferol, had little effect on these biochemical parameters. In rachitic growth plates, the uronic acid content was half that in normal growth plates, whereas ALPase activity was 2.5 times that in normal growth plates. Administration of 1,25-dihydroxycholecalciferol at a low dose (0.1 micrograms per kg of body weight) to rachitic rats increased the uronic acid content 1.4-fold and decreased ALPase activity by 40%. This compound, like 24,25-dihydroxycholecalciferol (10 micrograms per kg of body weight), increased the calcium level of the blood. However, administration of 24,25-dihydroxycholecalciferol had little effect on the uronic acid and ALPase contents in growth plates. These observations suggest that 1,25-dihydroxycholecalciferol is a bioactive form of vitamin D that plays an important role in the control of chondrocyte terminal differentiation.

The effects of histone subfractions on rat liver thyroid hormone receptor–DNA interaction were examined using an in-vitro DNA-cellulose binding assay. H1 histones bound to DNA showed reversible and potent inhibition of receptor–DNA binding without affecting receptor–hormone binding. Poly-lysine, bovine serum albumin, ovalbumin and cytochrome cdid not alter receptor–DNA binding. H1 histone subfractions (calf thymus lysine-rich histone (CTL)-1, CTL-2 and CTL-3) showed potent inhibition of receptor–DNA binding indistinguishable from each other. The quantity of H1 histone subfractions bound to DNA was the same. Although each subfraction has different functional properties, inhibition of receptor–DNA binding was a common feature of all the H1 histone subfractions, which is important for the non-random distribution of the receptor in chromatin.Binding of the receptor to core histones was investigated; it was found to bind to core histones more potently than to other proteins (H1 histone, ovalbumin and cytochrome c). Among core histone subfractions, H4 histone bound to the receptor most potently and is the candidate to be one of the acceptor sites of the receptor in chromatin.Journal of Endocrinology(1989) 121,337–341

An aqueous two-phase partitioning study of partially purified nuclear thyroid hormone receptor from rat liver was performed. Stability of 3,5,3′-tri-iodo-l-thyronine (T3)–receptor complex and T3-binding activity in the presence of dextran or polyethylene glycol were assessed in order to determine the amount of occupied or unoccupied receptors in each phase. Partition coefficients were calculated as the ratio of receptor concentration in the upper polyethylene glycol-rich phase H2O and that in the lower dextranrich phase H2O. The partition coefficient was a sensitive function of the salt at pH above 6·1 and below 5·1. The salt had no effect on the partition coefficient at pH around 5·6. These results suggest that the isoelectric point of the thyroid hormone receptor is about 5·6, confirming previous determinations using isoelectric focusing. The partition coefficient of the receptor decreased upon T3binding, regardless of the salt composition. In contrast, the partition coefficient of thyroxine-binding globulin increased upon T3binding. Free T3preferentially partitioned into the upper polyethylene glycol-rich phase and gave a partition coefficient higher than 1·0. These results strongly suggest that the decrease in the partition coefficient of the receptor upon hormone binding reflects conformational changes or changes in electrostatic properties of the receptor upon hormone binding. Such an alteration may be involved in biological activation of the receptor upon hormone binding.J. Endocr. (1988) 119,431–437

Summary Pharmacokinetic studies with orally administered 14C-oxyfedrine were carried out in rats and also in men.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

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ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.

The pharmacokinetics of cefotaxime were studied in 13 patients on haemodialysis or continuous ambulatory peritoneal dialysis (CAPD). One gram of cefotaxime was administered intravenously or intraperitoneally. In the haemodialysis patients, the serum concentration after 4 h was 9.4 mg/l and the T½ was 1.31 h. The 6-h value and T½ in the CAPD (intravenous) patients were 4.5 mg/1 and 1.59 hours, while the peak value in the dialysate was 11.6 mg/1. In the CAPD (intraperitoneal) patients, the cefotaxime concentration in the dialysate was 157 mg/l after 6 h and the peak value in the serum was 9.1 mg/l. Since cefotaxime is easily removed from the blood by both the haemodialysis and CAPD methods, accumulation of the drug in patients undergoing dialysis is unlikely to occur.

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