Nitric oxide prevents IL-1beta and IFN-gamma-inducing factor (IL-18) release from macrophages by inhibiting caspase-1 (IL-1beta-converting enzyme)

Procytokine processing by caspase-1 is required for the maturation and release of IL-1beta and IFN-gamma-inducing factor (IGIF) (or IL-18) from activated macrophages (Mphi). Nitric oxide (NO) has emerged as a potent inhibitor of cysteine proteases. Here, we tested the hypothesis that NO regulates cytokine release by inhibiting IL-1beta-converting enzyme (ICE) or caspase-1 activity. Activated RAW264.7 cells released four to five times more IL-1beta, but not TNF-alpha, in the presence of the NO synthase inhibitor N(G)-monomethyl-L-arginine. Stimulated peritoneal Mphi from wild-type mice (inducible NO synthase (iNOS)+/+) also released more IL-1beta if exposed to N(G)-monomethyl-L-arginine, whereas Mphi from iNOS knockout mice (iNOS-/-) did not. Inhibition of NO synthesis in stimulated RAW264.7 cells also resulted in a threefold increase in intracellular caspase-1 activity. The NO donor S-nitroso-N-acetyl-DL-penicillamine inhibited caspase-1 activity in cells as well as the activity of purified recombinant caspase-1 and also prevented the cleavage of pro-IL-1beta and pro-IGIF by recombinant caspase-1. The inhibition of caspase-1 by NO was reversible by the addition of DTT, which is consistent with S-nitrosylation as the mechanism of caspase-1 inhibition. An in vivo role for the regulation of caspase-1 by NO was established in iNOS knockout animals, which exhibited significantly higher plasma levels of IL-1beta and IFN-gamma than their wild-type counterparts at 10 h following LPS injection. Taken together, these data indicate that NO suppresses IL-1beta and IGIF processing by inhibiting caspase-1 activity, providing evidence for a unique role for induced NO in regulating IL-1beta and IGIF release.     

Estimation of changes in single muscle fibre diameter in different solutions by diffraction studies

Studies of the effects of varying tonicity on the contraction of muscle fibres requires a knowledge of the corresponding changes in fibre diameters, if the observed mechanical changes are to be attributed to various factors, including that of the interfilamental spacing. Hitherto, measurements of such changes in fibre diameters involved tedious measurements of the diameters of the fibres at numerous points along its length. We now describe a technique whereby the change in fibre diameter can be deduced from measurements of the corresponding intensity ratio of laser diffraction lines when the fibre is bathed in solutions of differing tonicities.     

Precipitated immune complexes of IgM induce the generation of reactive oxygen species by rabbit polymorphonuclear leucocytes

We report that immune complexes of IgM (ICIgM) antibodies and ovalbumin in the form of a precipitate from the equivalence zone induce the generation of reactive oxygen species by rabbit blood polymorphonuclear leucocytes (PMN), as measured by the chemiluminescence (CL) production in the presence of luminol. The kinetics of CL generation induced by ICIgM is quite different from that induced by precipitated immune complexes of IgG (ICIgG): the maximum rate of CL production for ICIgM occurs around 14 min, whereas for ICIgG it occurs about 5 min after incubation with the cells. Also the triggering of the process requires a higher concentration of ICIgM than of ICIgG. Evidence is presented that these effects are not mediated by interaction of the antigen (ovalbumin) with the cell, since immune precipitates of ovalbumin and the F(ab')2 fragment had no effect. Our observations that precipitated ICIgM can also be an effective stimulus for CL generation and thus for O2- production reveal a new functional capability of PMN. These results may have implications for the understanding of the participation of ICIgM (as well as of ICIgG) in inflammatory reactions mediated by PMN in immune complex diseases, and in the mechanisms of defense against microbes and other non-self agents.     

Prospective controlled randomized study of in vitro fertilization versus intracytoplasmic sperm injection in the treatment of tubal factor infertility with normal semen parameters

We report a case of aortitis syndrome, in which carotid ultrasonography was a useful approach for the diagnosis. A 21-year-old woman was admitted to our hospital for persistent fever. No specific physical findings or laboratory abnormalities were observed except high fever and marked increase of erythrocyte sedimentation rate and c reactive protein. Since clinical trial of antibiotics and antituberculosis agents resulted in no effectiveness, prednisolone was started, but the effect was limited. After that, she complained of the neck pain, and vascular murmur became apparently audible at the pain site. Ultrasonography of the carotid artery revealed the smooth lumen and homogeneous, non-hyperechoic intimal thickening. Then aortography confirmed the diagnosis of aortitis syndrome. Cyclophosphamide combined with steroid therapy diminished the disease activity. In this case, carotid ultrasonography gave us the important information to enforce the aortography. If ultrasonography of the carotid artery has been popularized for aortitis syndrome, and its findings have been standardized, non-invasive diagnosis of this disease will be taken a step forward.