The effect of low sulfate concentrations on the glycosaminoglycan synthesis in anatomically intact articular cartilage of the mouse

We have studied the effect of environmental sulfate concentration on the glycosaminoglycan synthesis of anatomically intact patellar cartilage of the mouse in vitro. Incubation of mouse patellae in medium with sulfate concentrations below 0.5 mM resulted in a diminished incorporation of sulfate but in unaltered incorporation of glucosamine. This suggested the synthesis of undersulfated glycosaminoglycans under these conditions. We characterized glycosaminoglycans synthesized at three different sulfate concentrations: a sulfate concentration physiological for the mouse (1.0 mM), a sulfate concentration in the range where sulfate incorporation was strongly diminished (0.1 mM), and an extremely low sulfate concentration (10 nM). Analysis of glycosaminoglycan disaccharides and DEAE anion chromatography of the glycosaminoglycans could not confirm the synthesis of undersulfated glycosaminoglycans at 0.1 mM. The chromatogram of glycosaminoglycans synthesized in medium containing 10 nM showed the presence of a very low sulfated glycosaminoglycan pool not observed at higher medium sulfate concentrations. Intermediately sulfated glycosaminoglycans were also synthesized during incubation with 10 nM sulfate. So, our data indicate that only very low sulfate concentrations in the medium lead to the synthesis of undersulfated glycosaminoglycans and that the sulfation mechanism of murine patellar cartilage chondrocytes does not seem to fit completely in an "all-or-nothing" pattern.     

Mitochondrial Antibodies in Primary Biliary Cirrhosis: II. The Complement Fixing Antigen as a Component of Mitochondrial Inner Membranes

The autoantibodies found in the sera of patients with primary biliary cirrhosis have been shown to react with a component of the mitochondrial inner membranes. Outer membranes were inactive. The purity of the inner and outer membrane fractions obtained by 2 different methods was assessed by electron microscopy and marker enzyme tests. Using negative-staining it was possible to visualize antibody binding to mitochondrial membranes. At high resolution it could be seen that the 90° particles on the cristal membranes were not involved in the reaction with antibody, but it was not possible to establish in the present studies the precise antigenic site upon the mitochondrial inner membranes.     

Human smooth pursuit during transient perturbations of predictable and unpredictable target movement

Summary The predictive component of human smooth pursuit was studied by perturbing sinusoidal target motion at unpredictable instants. The disturbances consisted of either a brief period of stabilization of the target on the fovea or a replacement of the sine by a ramp displacement for half a period. To minimize the effects of a possible change of the tracking strategy by the subject the transitions were masked and only the initial part of the response to the disturbance was analyzed. After stabilization on the fovea the eye oscillation continued at the frequency of the preceding target movement for about one half-cycle, whereupon the oscillation was rapidly damped. The mean unidirectional smooth eye acceleration was 70% of the mean unidirectional target acceleration prior to the stabilization. This suggests that during pursuit of a sinusoidal target movement about 75% of the oculomotor response is generated by predictive processes. When the sine was replaced by a ramp, starting at the velocity zero-crossing, the eye accelerated away from the target for ca. 180 ms irrespective of the frequency of prior tracking. In contrast, when the ramp started at the peak velocity of the sinusoidal target motion the eye accelerated away from the target for more than a quarter period. After foveal stabilization during pursuit of a pseudorandom stimulus, the eye continued to oscillate for less than one period at approximately the highest frequency present in the stimulus. The frequency characteristics of human smooth pursuit of predictable as well as unpredictable target motion were correctly simulated by a model, which derived its predictive properties from a lead element, tuned to the current frequency of the target motion.     

High levels of Lp(a) lipoprotein in a family ith cases of severe pre-eclampsia

We report a family with two cases of severe pre-eclampsia/eclampsia in which very high levels of Lp(a) lipoprotein were found. The serum level of Lp(a) lipoprotein is genetically determined and the Lp(a) apolipoprotein has a close homology to plasminogen. Very high levels of Lp(a) lipoprotein might interfere with the fibrinolytic/thrombolytic process in man. A previous report suggested that a high maternal serum Lp(a) lipoprotein level can cause fetal growth retardation, and it is proposed that very high levels might lead to increased deposition of fibrin in the uterine spiral arteries in pregnancy, which is central in the pathogenesis of pre-eclampsia. If confirmed, a very high Lp(a) lipoprotein level could be one risk factor for pre-eclampsia that is genetically determined.     

Role of interleukin 1 in antigen-induced exacerbations of murine arthritis.

The mechanism underlying the chronic and intermittent course of rheumatoid arthritis is not elucidated. In the present study, the role of interleukin 1 (IL-1) was investigated in exacerbations of antigen-induced arthritis in mice. A flare-up of smoldering inflammation (weeks 3 to 4 of antigen-induced arthritis) was inducible by injection of a small amount of methylated bovine serum albumin into the hypersensitive knee joint. Immunohistochemistry showed IL-1 expression in the synovial lining layer and in focal areas of the inflamed synovium during the flare-up. IL-1 was also measured in 1-hour culture supernatant of synovial tissue taken during the flare-up by a bioassay. The expression of both immunoreactive and bioactive IL-1 in the hypersensitive joint peaked around 6 hours after antigen (2 micrograms of methylated bovine serum albumin) injection and declined thereafter. Antigen rechallenge induced an acute joint swelling of the arthritic joint but not in the naive joint of the sensitized mouse, yet synovia of both joints produced IL-1 after antigen injection. Remarkably, a single intravenous injection of rabbit anti-IL-1 alpha and -beta antibodies 1 hour before antigen rechallenge neutralized IL-1 in the joint. Anti-IL-1 treatment significantly reduced the antigen-induced joint swelling (30 to 40%) but did not affect the profound influx of polymorphonuclear cells in the onset of the exacerbation. However, a profound relief of the inflammation (synovitis) was obtained by IL-1 blockade on day 4 of the exacerbation. Chondrocyte proteoglycan synthesis was markedly suppressed in the antigen-challenged naive knee joints suggesting that this was a direct IL-1 effect as the inflammation was insignificant. Anti-IL-1 treatment was able to maintain chondrocyte proteoglycan synthesis in the antigen-rechallenged joint, which was highly suppressed in the control group. Furthermore, the enhanced proteoglycan breakdown in the antigen-rechallenged joints was significantly decreased in the anti-IL-1 group. We concluded that IL-1 is an important mediator in exacerbations of murine arthritis, and amelioration of cartilage pathology was obtained with anti-IL-1 antibody treatment.     

Adoptive transfer of T lymphocytes to T-cell-depleted mice inhibits Escherichia coli translocation from the gastrointestinal tract.

Bacterial translocation is defined as the passage of viable bacteria from the gastrointestinal (GI) tract to extraintestinal sites, such as the mesenteric lymph node (MLN), spleen, liver, kidneys, and blood. Previously, we reported that depletion of CD4+ and/or CD8+ T cells promotes bacterial translocation from the GI tract to the MLN. In the present study, CD4+ and/or CD8+ T cells, harvested from donor mice, were adoptively transferred to mice previously depleted of T cells by thymectomy plus intraperitoneal injection of rat anti-mouse T-cell monoclonal antibodies. The adoptively transferred CD4+ and/or CD8+ T cells inhibited the translocation of Escherichia coli from the GI tract. Migration of the adoptively transferred T cells to the spleens and MLNs of the recipient mice was determined by utilizing Thy 1.1+ donor cells adoptively transferred into mice whose cells express the Thy 1.2 marker. These results provide further evidence of the importance of T cells in the host immune defense against bacterial translocation from the GI tract.