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PubMed1988

The effects of amrinone, cyclophosphamide and anti-platelet serum on platelet production in the Gunn rat.

The effect of amrinone on platelet production was differentiated from that of a known bone-marrow cytotoxic agent (cyclophosphamide) and anti-platelet serum (APS). The rate of platelet production has been observed over a 4-day period in the Gunn rat using [75Se]selenomethionine cohort labelling of platelets following administration of either amrinone, 160 mg kg-1 day-1, cyclophosphamide, 30 mg kg-1 day-1 or APS, 0.75 ml. Platelet numbers were reduced by amrinone, cyclophosphamide and APS. The rate of platelet production was increased following APS and suppressed by cyclophosphamide, but the rate of platelet production when expressed as the selenomethionine incorporation in counts per minute (cpm) per 10(8) platelets appeared to be increased in amrinone-treated animals. When these values are expressed as radioactivity per unit platelet volume the difference between the control and the amrinone-treated group was reduced but the difference between the control, APS- and cyclophosphamide-treated groups remained unchanged. It is concluded that in the Gunn rat amrinone affects platelet production by producing fewer, larger platelets.

Amrinone↗
PubMed1996

Multiple forms of selenoprotein P in rat plasma.

SDS-PAGE of immunoaffinity-purified rat seleno-protein P demonstrates a major band at 57 kDa and a less intense band at 45 kDa, suggesting the existence of more than one form of the protein. Separate experiments were carried out in which plasma from rats administered 75Se and immunoaffinity-purified 75Se-labeled selenoprotein P were applied to a heparin-Sepharose column at pH 7. In each experiment three peaks of 75Se-labeled protein were eluted during a continuous gradient from pH 7 to pH 8.5. The remaining 75Se-labeled material was eluted as a single peak by 1 M NaCl. Phosphorimaging of the SDS-PAGE gel of the peaks revealed that the first peak (1a) contained two bands (45 and 57 kDa) while the three remaining peaks each contained one band (peak 1b, 45 kDa and peaks 2 and 3, 57 kDa). N-terminal sequencing of the first eight amino acids of each band revealed that all five have the same N-terminal amino acid sequence and therefore are forms of selenoprotein P. Staining using a digoxigenin-based staining procedure revealed that all five forms contain carbohydrate. This demonstrates that plasma-derived selenoprotein P can be separated into five forms based on SDS-PAGE migration and heparin-Sepharose affinity and that at least two isoforms of selenoprotein P exist based on the presence of 45- and 57-kDa forms. The five forms have been given designations based on their order of elution from the heparin-Sepharose column and their M(r) on SDS-PAGE: selenoprotein P57A (peak 1a, 57 kDa), selenoprotein P45A (peak 1a, 45 kDa), selenoprotein P45B (peak 1b, 45 kDa), selenoprotein P57B (peak 2, 57 kDa), and selenoprotein P57C (peak 3, 57 kDa).

Amino Acid Sequence↗
PubMed1998

Selenoproteins are expressed in fetal human osteoblast-like cells.

Selenoproteins are involved in mechanisms of cell differentiation and defense. We investigated the expression of glutathione peroxidases, as well as other selenoproteins, in fetal human osteoblasts (hFOB-cells). Using 75-selenium metabolic labelling of viable hFOB-cells, we identified several selenoproteins in cell lysates of about 45-80 kDa and in the migration range of 14 kDa to 24 kDa. Cells expressed low mRNA levels of both cellular glutathione peroxidase and plasma glutathione peroxidase mRNA as analysed by Southern analysis of RT-PCR products. Basal cellular glutathione peroxidase enzyme activity in hFOB-cells (19.7 nmol NADPH oxidised per min and microg protein) was further increased 2.5-fold by the addition of 100 nM sodium selenite to the culture medium for 3 days. Furthermore, expression of selenoprotein P mRNA was demonstrated by RT-PCR. hFOB-cells did not show activities of the selenoproteins type I or type II 5'-deiodinase. In summary, we identified cellular glutathione peroxidase, plasma glutathione peroxidase and selenoprotein P among of a panel of several 75-selenium labelled proteins in human fetal osteoblasts. The expression of selenoproteins like glutathione peroxidases in hFOB-cells represents a new system of osteoblast antioxidative defense that may be relevant for the protection against hydrogen peroxide produced by osteoclasts during bone remodelling.

Antioxidants↗
PubMed1999

Selenocysteine-containing thioredoxin reductase in C. elegans.

Mammalian thioredoxin reductases contain a TGA-encoded C-terminal penultimate selenocysteine (Sec) residue, and show little homology to bacterial, yeast, and plant thioredoxin reductases. Here we show that the nematode, Caenorhabditis elegans, contains two homologs related to the mammalian thioredoxin reductase family. The gene for one of these homologs contains a cysteine codon in place of TGA, and its product, designated TR-S, was previously suggested to function as thioredoxin reductase. The other gene contains TGA and its product is designated TR-Se. This Sec-containing thioredoxin reductase lacks a canonical Sec insertion sequence element in the 3'-untranslated area of the gene. TR-Se shows greater sequence similarity to mammalian thioredoxin reductase isozymes TR1 and TR2, whereas TR-S is more similar to TR3. TR-Se was identified as a thioredoxin reductase selenoprotein by labeling C. elegans with 75Se and characterizing the resulting 75Se-labeled protein by affinity and other column chromatography and gel-electrophoresis. TR-Se was expressed in Escherichia coli as a selenoprotein when a bacterial SECIS element was introduced downstream of the Sec TGA codon. The data show that TR-Se is the major naturally occurring selenoprotein in C. elegans, and suggest an important role for selenium and the thioredoxin system in this organism.

3' Untranslated Regions↗
PubMed1994

Gall-bladder and colonic retention of SeHCAT: a re-evaluation.

A number of suggested alternatives and emendations to the 7-day SeHCAT retention test have been compared with whole-body counting. It was found that correction for colonic retention is an unnecessary complication in patients with diarrhoea and that imaging either of the gall-bladder or of the distribution of activity in the intestines at 24 h does not add useful information to the standard 7-day retention measurement. Neither could the patterns of colonic uptake identified in patients following extensive ideal resection or radiotherapy be reproduced in patients with idiopathic diarrhoea. Sadly it must be concluded that neither of these shortened techniques is of clinical value in patients with intact small bowel and that there remains no reliable alternative to the 7-day Se HCAT retention test. The simplest technique the best.

Bile Acids and Salts↗
PubMed1978

Correlation of electron microscopic and secretory response of human parathyroid adenomas with different calcium concentrations in organ culture.

Twelve parathyroid chief cell adenomas from patients with primary hyperparathyroidism were incubated in a tissue culture system in the presence of different calcium concentrations and for various time periods. The endocrine response of the tissue was examined electron microscopically and radioimmunologically. After incubation in a medium of low calcium concentration the parathyroid adenomas showed ultrastructural signs of stimulation with proliferation of the hormone-synthesizing organelles. The development of the ultrastructural response could first be observed after four hours and increased up to several days. Radioimmunologically, an increase of the hormone secretion could be demonstrated. Converse results were obtained after incubation of the tumor tissue under suppressive culture conditions. To check for de-novo synthesis of the hormone released the tissue was incubated in a 75Se-methionine-containing medium. This resulted in radioactivity of the secreted parathyroid hormone, indicating de novo synthesis in our culture system. The biological potency of the released hormone was demonstrated by comparison of the PTH out of the medium with the international human MRC standard using two different radioassays.

Adenoma↗
PubMed1990

A black adrenocortical adenoma causing Cushing's syndrome not imaged by radiocholesterol scintigraphy.

In a 33-year-old female patient with left adrenal tumour and Cushing's syndrome, adrenocortical scintigraphy with radiocholesterol did not image the tumour nor the suppressed contralateral gland. Histology showed a black adrenocortical adenoma composed only of compact cells; there was no evidence of malignancy. This demonstrates that non-visualization of the adrenal glands in a patient with Cushing's syndrome is not invariably due to adrenal carcinoma. The literature on black adrenal adenomas causing Cushing's syndrome is reviewed.

Adenoma↗
PubMed1990

Radiolabeling of Necator americanus third stage larvae with 75Se-methionine.

Third stage larvae of Necator americanus were radiolabeled with 75Se-methionine by two methods. Larvae labeled in aqueous cultures contained 188 and 25 cpm per sheathed and ex-sheathed larva, respectively. Larvae labeled in coproculture incorporated 25 and 18 cpm per sheathed and ex-sheathed larva, respectively. All of the label was decayed in 5 days from larvae labeled in aqueous cultures, whereas appreciable amounts of radioactivity were still detectable at day 7 of chase period in coproculture labeled larvae.

Animals↗
PubMed1995

Uptake of 75-Selenium into the central nervous system of the rat.

These experiments have investigated selenium movement between blood and the CNS in anaesthetized rats. Each animal was anaesthetized and the left femoral blood vessels cannulated for blood withdrawal and solute infusion. Each rat received 75-Se as sodium selenite infused in normal saline and experiments lasted between 5 minutes and 5 hours during which blood samples were periodically taken. At termination, the CNS was removed, dissected and analysed with the plasma samples for 75-Se radioactivity by gamma-counting. Data were analyzed by multiple-time uptake analysis. Results showed unidirectional uptake of 75-Se into the CNS and some regional differences were found. On average the CNS influx rate constant (Kin) was about 7 +/- 1 x 10(-5) ml/min/g. This indicates that the 75-Se most likely entered the CNS in a protein-bound form.

Animals↗
PubMed1977

In vivo study of platelet kinetics by 75Se-methionine in different haematological disorders.

Labeling of platelets in vivo by 75Se-methionine was performed in premalignant and malignant haematological disorders for evaluation of the kinetics of platelet maturation. The "normal" platelet maturation time (time between the injection of the isotope and maximum radioactivity of separated blood platelets) in eight non-haematological patients showing normal platelet counts was 9.1 days. A shortening of platelet maturation time of 5-7 days was observed in three of four cases with panmyelopathy (high bone marrow cellularity), in three of four cases with malignant lymphatic disorders (multiple myeloma, chronic lymphocytic leukaemia, lymphosarcoma), and in two of four cases with myeloproliferative syndromes. No correlation to the peripheral platelet counts was observed. For explanation of the premature platelet release from the bone marrow a disturbance of the megakaryocyte maturation is suggested.

Adult↗
PubMed1992

Measurement of bile acid half-life using [75Se]HCAT in health and intestinal diseases. Comparison with [75Se]HCAT abdominal retention methods.

The diagnostic values in detecting terminal ileum dysfunction using [75Se]HCAT have been assessed for two different scintigraphic techniques in 58 subjects. The measurement of [75Se]HCAT T1/2 in the enterohepatic circulation by daily gallbladder scintigraphy showed 78% sensitivity, 96% specificity, 96% positive predictive value, and 78% negative predictive value at the optimal cutoff level of 2.0 days; lower--but not significantly different--figures were observed for [75Se]HCAT total abdominal retention four and seven days after isotope administration, at the optimal cutoff levels of 40% and 22%, respectively. [75Se]HCAT T1/2 was then evaluated in 60 patients with various intestinal diseases. Sixty-nine percent (9/13) of patients with diarrhea of obscure origin showed abnormal [75Se]HCAT T1/2. Bile acid malabsorption using [75Se]HCAT can be investigated by the noninvasive measurement of its enterohepatic T1/2 and may play a pathogenetic role in patients with diarrhea of obscure origin.

Adolescent↗