Plasma proteins immunologically related to inter-alpha-trypsin inhibitor.

SDS-polyacrylamide gel electrophoresis and immunoblot were applied to analysis of plasma proteins immunologically related to inter-alpha-trypsin inhibitor (ITI). In this system, anti-ITI sera were able to identify ITI and other components with an Mr near 120 kDa which would be degradation products of ITI by limited proteolysis. An anti-UTI (urinary trypsin-inhibitor) serum could detect, beside these derivatives, two minor components (Mr values near 90 and 60 kDa). Analysis of perchloric acid supernatants of plasma samples, using the same technic, induced visualization of a new component, similar to urinary trypsin inhibitor which could not be detected by direct analysis. This one was also characterized in a higher content in pathological samples (renal failure and infectious diseases).

Effect of Cleome arabica leaf extract, rutin and quercetin on soybean lipoxygenase activity and on generation of inflammatory eicosanoids by human neutrophils.

The effects of Cleome arabica leaf extract, rutin and quercetin on soybean lipoxygenase (Lox) activity and on calcium ionophore (A23187)-stimulated generation of the leukotriene B4 and prostaglandin E2 by human neutrophils were examined. The extract (25 microg/ml), rutin (25 microM) and quercetin (25 microM) inhibited LTB4 synthesis at all concentrations of A23187 used. The extract at 1-100 microg/ml and rutin at 1-100 microM inhibited LTB4 generation by neutrophils stimulated with 1 microM A23187 by about 50%. PGE2 production in response to different concentrations of A23187 was affected in a biphasic manner by the extract and rutin. Quercetin at 1-100 microM caused concentration-dependent inhibition of LTB4 and PGE2 production. The extract, rutin and quercetin caused concentration-dependent inhibition of soybean Lox activity. These results indicate that rutin, quercetin and an extract of C. arabica containing these compounds inhibit Lox activity, consequently decreasing LTB4 production. Thus, these compounds or extracts containing them may be beneficial for the treatment of inflammatory conditions, particularly those characterised by excessive leukotriene generation.

The effect of the glycosaminoglycan chain removal on some properties of the human urinary trypsin inhibitor.

The major urinary trypsin inhibitor UTI I is a proteoglycan. UTI c (Mr 26,000), produced by chrondroitin lyase digestion of UTI I, was isolated and characterized. About 90% of the glycosaminoglycan chain was removed by this treatment without proteolytic modification, as assessed by amino-acid composition and N-terminal sequence of UTI c. Its electrophoretic mobilities on alkaline and SDS-PAGE are identical with those of UTI II which occurs in urine during storage. To study the role of the glycosaminoglycan chain on the inhibitory properties of UTI I, UTI I and UTI c were compared using different proteinases as target enzymes. The inhibitory activity towards bovine trypsin and chymotrypsin as well as human granulocytic cathepsin G did not differ significantly. However, towards human granulocytic elastase, the equilibrium dissociation constant (Ki) is 5 times higher for UTI c than for UTI I. Weak inhibitory activities were measured on human plasmin, UTI c being more efficient than UTI I. The acid-stability of UTI I is not modified after chrondroitin lyase treatment. UTI I and UTI c are equally sensitive to trypsinolysis indicating that the covalently bound glycosaminoglycan chain does not play an important role for the stability of UTI I.

Effects of flavonols on the generation of superoxide anion radicals by xanthine oxidase and stimulated neutrophils.

The effects of three aglycon flavonols (myricetin, quercetin, and kaempferol) and the natural glycoside rutin on superoxide anion radical generating systems were investigated. Quercetin, myricetin, and kaempferol inhibited the formation of uric acid from xanthine by xanthine oxidase, while rutin was ineffective. The generation of superoxide anion radicals by this system was determined by either reduction of cytochrome c or Pholasin luminescence. A scavenging of superoxide was only observed for myricetin and to a small extent for rutin. All flavonols tested inhibited the Pholasin luminescence of fMet-Leu-Phe-stimulated neutrophils. Rutin influenced the oxidative burst of neutrophils in the same way as wortmannin and LY294002, two inhibitors of the phosphoinositide 3-kinase gamma. Indeed, rutin inhibited the activity of this enzyme, whereas the three other flavonols showed no effect. Thus, an inhibition of enzymes involved in signaling rather than a scavenging of superoxide anion radicals dominates in fMet-Leu-Phe-stimulated neutrophils exposed to flavonols.

[Introduction to a study of specific antibodies (IgG and IgM) detection with ELISA in the diagnosis of poliomyelitis]. / Introduction à une étude sur la détection des anticorps spécifiques (IgG et IgM) par ELISA dans le diagnostic de la poliomyélite.

Samples of single sera collected from 38 patients with different clinical diagnosis were studied in order to perform ELISA techniques with the purpose of detecting poliomyelitis IgG and IgM antibodies. The résults were compared through antibody titration by neutralization test. 21 pairs of sera from infants suffering from acute flaccid paralysis were studied by ELISA-IgM, ELISA-IgG and neutralization test. Stool samples were collected from 20 of the latter patient. Wild poliovirus type 1 was isolated in 8 cases. ELISA-IgM technique was positive in 14 cases. The true positive poliomyelitis diagnosis was based on the persistence of flaccid paralysis 60 days after the onset and on wild poliovirus isolation with significant increase in antibody level. 16 cases were classified as poliomyelitis, 2 cases as non poliomyelitic paralysis and 3 cases as undetermined. 16 out of the 18 well established diagnosis were in agreement (88.8%) with the detection or not of IgM antibodies by ELISA. The specificity of these IgM ELISA antibodies was examined by studying 11 cases of lymphocytic meningitis. Cross reaction in serological responses between polioviruses and coxsackieviruses was observed. These cross reactions should be evaluated by studying a greater number of cases. The poliovirus ELISA-IgM is a sensitive, economical and rapid method to be used in poliomyelitis diagnosis to complete the neutralizing test and virus isolation.