The effects of monocyte-conditioned medium and interleukin 1 on the synthesis of collagenous and non-collagenous proteins by mouse bone and human bone cells in vitro

Cultural adherent human mononuclear cells produce factor(s) which stimulate the release of calcium from new-born mouse calvaria in organ culture. This stimulation of bone resorption is accompanied by an inhibition of the incorporation of [³H]proline into collagen which is independent of increased prostaglandin production by the bone. When human osteoblast-like cells are treated with conditioned medium from human mononuclear cells, collagen accounts for a decreased proportion of the protein synthesised. This effect on matrix synthesis is not accompanied by an inhibitory action of the monocyte-conditioned medium preparations on net cell proliferation. In human osteoblast-like cell cultures, partially purified human interleukin 1 also inhibits the production of the bone-specific protein osteocalcin in a dose-dependent fashion. These observations are consistent with the hypothesis that products of human monocytes similar to, or identical with, human interleukin 1 may be important regulators of bone metabolism and may contribute to the bone loss seen in diseases such as chronic rheumatoid arthritis.     

Class I-like MHC molecules expressed by baboon placental syncytiotrophoblast

Human placental villous trophoblast is known to be unreactive with W6/32 and other monoclonal antibodies recognizing monomorphic determinants of human class I MHC heavy chains, whereas extravillous cytotrophoblast in the placental bed is W6/32-reactive by immunohistology. We have now demonstrated, in contrast, that syncytiotrophoblast is the only cellular component of baboon early placental villous tissue which is reactive with any of these antibodies. Radioimmunoprecipitation of detergent-solubilized baboon placental membrane preparations, and subsequent SDS-PAGE, has shown the W6/32-reactive component to have an m.w. of 41,000 and to be associated with beta 2-microglobulin, whereas baboon peripheral lymphocytes express 45,000 m.w. W6/32-reactive antigens comparable with the HLA-A,B,C heavy chains of human lymphocytes.     

Antithrombin Glasgow, 393 Arg to His: a P1 reactive site variant with increased heparin affinity but no thrombin inhibitory activity

Antithrombin Glasgow is a hereditary abnormal antithrombin that has lost thrombin inhibitory activity. It was isolated from the plasma of a 41-year-old male with a history of thrombotic events. Antithrombin Glasgow was purified from plasma using heparin-Sepharose chromatography at pH 7.4 eluting with increasing concentrations of NaCl. The normal protein eluted with 0.9 mol/l NaCl and Glasgow with 1.05 mol/l NaCl. Electrophoresis in agarose at pH 8.6 showed the variant to migrate more anodally than normal. The C-terminal small fragment resulting from catalytic cleavage with elastase between P3 and P4 of the reactive loop was isolated and sequenced. This showed the replacement of the arginine at residue 3 by a histidine. This is residue 393 in the intact molecule. The findings suggest that heparin, on binding, interacts indirectly with the reactive centre region of antithrombin.     

Use of the true absorption coefficient as a measure of bioavailability of radiocaesium in ruminants

Limitations of existing methods to describe the bioavailability of dietary radionuclides to ruminants (the transfer coefficient and apparent absorption coefficient) have led to the alternative suggestion of using the true absorption coefficient (A _t). Various approaches to estimatingA _t for radiocaesium, involving the intravenous administration of a second isotope, are presented and discussed with reference to results from studies in which a range of radiocaesium sources were examined in sheep. Although estimates ofA _t differed between the sources, they were reasonably consistent between measurement techniques. Those methods which involved the estimation of endogenous faecal excretion of radiocaesium could be used with previously contaminated animals and did not require continuous administrations of radiocaesium isotopes, but gave unreliable results for sources of low bioavailability. Methods based on estimating the turnover rate of dietary radiocaesium through blood plasma were sufficiently sensitive to measureA _t for the range of sources studied. However, they require previously uncontaminated animals and continuous administration of both isotopes for approximately 7 days. Bioavailability is more effectively measured asA _t than as the transfer or apparent absorption coefficients sinceA _t does not incorporate factors relating to the metabolism of radiocaesium in the tissues of the animal. The results of these studies show that differences in transfer coefficients between sheep and cattle and between sheep of differing ages are not due to variation in absorption across the gut. The potential for applying these approaches to other radioactive elements is discussed.     

Independent cerebral vasoconstrictive effects of hyperoxia and accompanying arterial hypocapnia at 1 ATA.

Breathing 100% O2 at 1 atmosphere absolute (ATA) is known to be associated with a decrease in cerebral blood flow (CBF). It is also accompanied by a fall in arterial Pco2 leading to uncertainty as to whether the cerebral vasoconstriction is totally or only in part caused by arterial hypocapnia. We tested the hypothesis that the increase in arterial Po2 while O2 was breathed at 1.0 ATA decreases CBF independently of a concurrent fall in arterial Pco2. CBF was measured in seven healthy men aged 21-62 yr by using noninvasive continuous arterial spin-labeled-perfusion MRI. The tracer in this technique, magnetically labeled protons in blood, has a half-life of seconds, allowing repetitive measurements over short time frames without contamination. CBF and arterial blood gases were measured while breathing air, 100% O2, and 4 and 6% CO2 in air and O2 backgrounds. Arterial Po2 increased from 91.7 +/- 6.8 Torr in air to 576.7 +/- 18.9 Torr in O2. Arterial Pco2 fell from 43.3 +/- 1.8 Torr in air to 40.2 +/- 3.3 Torr in O2. CBF-arterial Pco2 response curves for the air and hyperoxic runs were nearly parallel and separated by a distance representing a 28.7-32.6% decrement in CBF. Regression analysis confirmed the independent cerebral vasoconstrictive effect of increased arterial Po2. The present results also demonstrate that the magnitude of this effect at 1.0 ATA is greater than previously measured.     

Insulin-secreting beta-cells possess specific receptors for interleukin-1 beta

The effect of the cytokine interleukin-1 beta on the insulin secretory responsiveness of single beta-cells (HIT-T15) was investigated. In the short-term, IL-1 beta induced a dosage-dependent stimulation of insulin release. In contrast, in the long-term, IL-1 beta, inhibited both basal and secretagogue-stimulated insulin secretion. We also demonstrate the simultaneous presence of specific high and low affinity binding sites for IL-1 beta on beta-cells. IL-1 beta, which has been implicated in the pathogenesis of insulin-dependent diabetes, may therefore mediate its opposing effects on beta-cells through a specific plasma membrane receptor.     

Polypeptide cross-linking in chloroplast membranes

Washed chloroplast membranes from romaine lettuce leaves were treated with the cross-linking reagent dimethyladipimidate (DMA) for various periods of time and subsequently analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Comparative examination of the electrophoretic profiles from control and treated membranes revealed that the light-harvesting chlorophyll-protein complex (LHCPC) was readily cross-linked to yield “dimers” and “oligomers” of higher molecular weight. Two polypeptides, of 25 and 23 kilodaltons, previously identified as two subunits of the LHCPC, were the major cross-linked species; other peptides were also cross-linked, but to a much lesser extent. These results suggest that cross-linking of chloroplast membranes with DMA, under our conditions, occurs primarily among the components of the LHCPC. We also measured the photosystem II activity in control and DMA-treated chloroplasts and found no impairment of this photochemical activity in the cross-linked chloroplasts as compared with controls.