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The thyroid plasma membrane contains a Ca(2+)-regulated NADPH-dependent H2O2-generating system which provides H2O2 for the peroxidase-catalysed biosynthesis of thyroid hormones. The electron transfer from NADPH to O2 catalysed by this system was studied by using diphenyleneiodonium (DPI), an inhibitor of flavo- and haemo-proteins. The prosthetic group of the H2O2 generator was removed by incubation with 5 mM CHAPS at 40 degrees C, and an active holoenzyme was reconstituted with FAD, but not with FMN. The H2O2-generating system also had an intrinsic Ca(2+)-dependent NADPH:ferricyanide reductase activity which is probably linked to its flavodehydrogenase component (or domain). Both activities, H2O2 production and ferricyanide reductase activity, were inhibited by DPI, with similar K1/2 (2.5 nmol/mg of protein). DPI only inhibited a system reduced with NADPH in the presence of Ca2+. NADPH could not be replaced by NADP+, NADH or sodium dithionite, suggesting the need for specific mild reduction of a redox centre in a particular conformation. Ferricyanide protected both activities against inhibition by DPI; the NADPH:ferricyanide reductase activity was completely protected at a ferricyanide concentration 20 times lower than that needed to protect the H2O2 formation, implying at least two target sites for DPI. One might be the flavodehydrogenase component; the other was beyond, on the entity which transfers the electrons to O2. This second site has not been identified.

In order to define the optimal dosage of a low molecular weight enoxaparine (Lovenox) in the prevention of clotting in extracorporeal circulation during hemodialysis, a multicentre trial was conducted in 72 patients dialysed in seven hemodialysis units. During three weeks, these patients received as antithrombic treatment a single injection of enoxaparine at the beginning of the session. The initial dose fixed by previous data concerning dialysis with high hemorrhagic risks patients was 0.5 mg/kg (50 U1 Anti-Xa/kg). According to the evaluation of thrombotic manifestations during a 4 hour dialysis, the dosage was progressively increased if necessary for each patient. For 41% patients, the initial dose of 0.5 mg/kg was maintained along the whole study; 59% patients needed higher dose, between 0.6 and 0.9 mg/kg. The mean dose for the whole patient population at the end of the study was 0.62 +/- 0.16 mg/kg. No complication nor side effect was noted. The influence of blood flow, nature of dialysis membrane, level of hematocrit was studied. In conclusion, 0.5 mg/kg of enoxaparine can prevent thrombotic manifestations in almost half of chronic hemodialysed patients with good results. Further studies could precise the place of personal or technical parameters in the choice of the optimal dose for each patient.

We describe a case of late perforation of the superior vena cava by a hemodialysis catheter inserted via the left internal jugular vein. This resulted in extravasation of blood. Malposition of the catheter was confirmed by CT scan. Removal of the catheter resulted in rapid resolution of symptoms. Confirmation of correct placement of central venous catheter must be obtained by chest X-ray contrast study.

Ceco-colonic myoelectrical activity was investigated in rats pretreated for 23 weeks by sennosides (10 or 40 mg/kg/day), Na-picosulfate (2.5 or 10 mg/kg/day) or laxative vehicle (control). On the last week of treatment the animals were equipped with Nichrome electrodes on the cecum, the proximal and distal colon. In comparison with controls, sennoside or Na-picosulfate treatment did not induce any significant (p > 0.05) change in the duration of long spike bursts (LSB) which are associated with phasic contractions. On the last 2 days of treatment the frequency of LSB for 2 h before and 2 h after laxative administration, as well as for 30 min after a 3-gram meal was not significantly (p > 0.05) different in control and treated animals. Similarly, on the first 2 days, as well as on days 13 and 14, after the end of treatment, no significant (p > 0.05) difference in the LSB frequency appeared between control and treated animals, in the fasted state or after a 3-gram meal. It is concluded that long-term treatment with sennosides or Na-picosulfate does not induce chronic changes in colonic motility in rats.

The effects of the trichotecene mycotoxin, deoxynivalenol, on gastric emptying and intestinal propulsion in mice and rats and gastrointestinal myoelectrical activity in rats were investigated. Gastric emptying and intestinal transit were evaluated after gavage with a milk meal containing a marker (51CrO4Na2) and radio-activity was counted in the stomach and 10 segments of the small intestine. The myoelectrical activity of the antrum, duodenum and jejunum was assessed by implanting electrodes for long-term electromyographic recordings. Deoxynivalenol given orally (50-1000 micrograms/kg) but not i.c.v. (5 micrograms/kg) 10 min before the test meal inhibited gastric emptying in a dose-related manner. Intestinal propulsion was reduced for the highest dose (1000 micrograms/kg) only. The inhibition of gastric emptying induced by deoxynivalenol was antagonized by ondansetron and granisetron given s.c. (50 micrograms/kg) but not by ondansetron i.c.v. (10 micrograms/kg). Metoclopramide, domperidone (1 mg/kg s.c.), methysergide, ritanserin and cisapride (2 mg/kg s.c.) did not modify the deoxynivalenol-induced inhibition of gastric emptying. In rats, gavage with a 2.5-ml milk meal increased the frequency of antral spike bursts from 1.9 +/- 0.9/min in the fasted state to 4.7 +/- 0.4/min and disrupted intestinal migrating motor complexes for 84.9 +/- 10.8 min. Oral administration of deoxynivalenol (50-100 micrograms/kg) 10 min before the meal did not modify the frequency of antral spike bursts but induced migrating motor complexes on the small intestine after the meal. This effect was reversed by ondansetron (10 micrograms/kg s.c.). It was concluded that, in rodents, deoxynivalenol inhibits gastric emptying by inducing intestinal migrating motor complexes through a peripheral action at the serotonin-3 receptors.

NADPH oxidation catalyzed by horseradish peroxidase is considerably increased by scopoletin and superoxide dismutase. These effects were used to develop a method for measuring H2O2 in a horseradish peroxidase, superoxide dismutase, and scopoletin system by measuring the NADPH oxidation rate. The optimal concentration of each reactant was determined. H2O2 could be detected and measured when it was present free in the medium or when it was produced by an H2O2-generating system, such as glucose-glucose oxidase or NADPH oxidase from thyroid plasma membranes. H2O2 was measured either by taking aliquots of the incubation medium or by placing NADPH directly in the medium and following the kinetics of NADPH oxidation. This latter approach required smaller amounts of biological material. In contrast to other methods, the H2O2 which is measured is regenerated. This method is 10 times more sensitive than the standard scopoletin method for H2O2 measurement and will detect a H2O2 production rate as low as 0.2 nmol per hour. The method is particularly suitable for biological systems in which small quantities of biological material are available.

The NADPH-dependent H2O2-generating system in a pig thyroid particulate fraction requires micromolar concentrations of Ca2+ for activity. The H2O2 generator could be Ca(2+)-desensitized (i.e. made fully active in the absence of Ca2+) by limited proteolysis with alpha-chymotrypsin or by treatment with ZnCl2. The Zn2+ effect was temperature- and dose-dependent with an apparent half-maximum concentration of 0.15 mM at 40 degrees C. Ca2+ desensitization was not reversed by adding the Zn2+ chelators, 1,10-phenanthroline and EGTA, but about one-third of the Ca(2+)-sensitivity was recovered after addition of 10 mM-dithiothreitol. The proteolysed enzyme and the Zn(2+)-treated enzyme had different Km values for NADPH. The Zn2+ effect did not seem to involve proteolysis or membrane fusion. These results indicate that Ca2+ regulation occurs via an autoinhibitory domain or inhibitory protein component of the H2O2-generator system. Its inhibitory effect may be removed by proteolysis or conformational changes, making the catalytic site accessible to the substrate NADPH and/or enabling electrons to be transferred from NADPH to O2.

A new case of pulmonary hemosiderosis with coeliac disease is reported. This is an extremely rare combination of which only nine instances have been published over the last 20 years. Three of the reported cases occurred in children. Apart from a marked predominance of males, the combination has no specific features. Firm evidence of a causal relationship between the two diseases is lacking but treatment with a gluten-free diet alone apparently had beneficial effects on the lung disease in two patients. Three pathogenic hypotheses are discussed herein: deposition of circulating immune complexes involving food allergens on the basement membrane of alveolar capillaries; reaction between antireticulin antibodies and an alveolar basement membrane antigen; or effect of adenovirus 12, a potential causative factor for celiac disease. Patients with idiopathic pulmonary hemosiderosis should routinely have tests for gluten intolerance, for instance a lactulose-mannitol intestinal permeability test. Lastly, other concomitant food allergies are reported.

Objective: To evaluate Diabeto, a computer-assisted diet education system., Research Design and Methods: One hundred five patients with insulin-dependent diabetes mellitus (IDDM) or non-insulin-dependent diabetes mellitus (NIDDM) were divided into two randomized groups to participate in the evaluation of Diabeto. With free access through Minitel, the French public videotex network, Diabeto helps diabetic patients self-monitor their diets and balance their meals with personalized counseling., Results: During the first 6-mo study, group A (54 patients) used Diabeto, whereas group B (51 patients) were control subjects. For the second 6-mo study, group B used the system. Evaluation was based on patients' dietetic knowledge, dietary habits, and metabolic balance., Conclusions: Diabeto led to a significant improvement of dietetic, knowledge in group A (P less than 0.0005) and also to improved dietary habits; decreased caloric intake in patients initially overeating (P less than 0.05), increase of dietary carbohydrate from 39.7 +/- 0.7 to 42.9 +/- 0.9% in patients with an initial intake less than 45% carbohydrate, and decrease of fat intake from 41.9 +/- 0.9 to 37.4 +/- 1.1% in patients with an initial intake of greater than 35% fat (P less than 0.0005). In the second study, in addition to similar improvements to those observed in the first study, HbA1 decreased from 11.0 +/- 0.4 to 9.9 +/- 0.4% (P less than 0.005) and fructosamine from 5.00 +/- 0.17 to 4.57 +/- 0.17% (P less than 0.001). Diabeto appears to be an effective therapeutic tool in the control of metabolic diseases.

The mechanism of NADPH oxidation catalyzed by horse-radish peroxidase (HRP) and 2,4-diacetyl-[2H]heme-substituted horse-radish peroxidase (DHRP) was studied. The roles of the different H2O2/peroxidase compounds were examined by spectral studies. The oxidized NADPH species were identified using the superoxide dismutase effect and by measuring the stoichiometry between NADPH oxidized and H2O2 used. In the presence of a mediating molecule, like scopoletin, both enzymes acted via a similar mechanism, producing only NADP degrees, which in turn reacted with O2 producing O2-. Consequently H2O2 was completely regenerated in the presence of superoxide dismutase and partially regenerated in its absence. In the absence of a mediating molecule, the H2O2 complex of both enzymes (compound I) catalysed NADPH oxidation by single-electron transfer, producing NADP degrees; compound II of these enzymes catalyzed NADPH oxidation more slowly by a direct two-electron transfer, producing NADPH+. There were difference between HRP and DHRP. HRP compound II was produced by the oxidation of 1 mol NADPH/mole compound I, while DHRP compound II was formed by the spontaneous conversion of compound I to compound II. The NADPH oxidation catalyzed by DHRP compound I did not lead to the formation of compound II. When H2O2 was produced slowly by the glucose/glucose-oxidase system, compound II was never formed and a pure O2- adduct of DHRP (compound III) accumulated.

The interaction of a fluorescent duplex DNA oligomer with the Klenow fragment of DNA polymerase I from Escherichia coli has been studied in solution by using time-resolved fluorescence spectroscopy. An aminonaphthalenesulfonate (dansyl) fluorescent probe was linked by a propyl chain to a C5-modified uridine base located at a specific site in the primer strand of the DNA oligomer. The fluorescent oligomer bound tightly to the Klenow fragment (KD = 7.9 nM), and the probe's position within the DNA-protein complex was varied by stepwise elongation of the primer strand upon addition of the appropriate deoxynucleoside triphosphates. The decay of the total fluorescence intensity and the polarization anisotropy were measured with a picosecond laser and a time-correlated single photon counting system. The fluorescence lifetimes, the correlation time for internal rotation, and the angular range of internal rotation varied according to the probe's position within the DNA-protein complex. These results showed that five or six bases of the primer strand upstream of the 3' terminus were in contact with the protein and that within this contact region there were differences in the degree of solvent accessibility and the closeness of contact. Further, a minor binding mode of the DNA-protein complex was identified, on the basis of heterogeneity of the probe environment observed when the probe was positioned seven bases upstream from the primer 3' terminus, which resulted in a distinctive "dip and rise" in the anisotropy decay. Experiments with an epoxy-terminated DNA oligomer and a site-directed mutant protein established that the labeled DNA was binding at the polymerase active site (major form) and at the spatially distinct 3'----5' exonuclease active site (minor form). The abundance of each of these distinct binding modes of the DNA-protein complex was estimated under solution conditions by analyzing the anisotropy decay of the dansyl probe. About 12% of the labeled DNA was bound at the 3'----5' exonuclease site. This method should be useful for investigating the editing mechanism of this important enzyme.

The thyroid plasma membrane contains a Ca2(+)-regulated NADPH-dependent H2O2 generating system which provides H2O2 for the thyroid peroxidase-catalyzed biosynthesis of thyroid hormones. The plasma membrane fraction contains a Ca2(+)-independent cytochrome c reductase activity which is not inhibited by superoxide dismutase. But it is not known whether H2O2 is produced directly from molecular oxygen (O2) or formed via dismutation of super-oxide anion (O2-). Indirect evidence from electron scavenger studies indicate that the H2O2 generating system does not liberate O2-, but studies using the modified peroxidase, diacetyldeuteroheme horseradish peroxidase, to detect O2- indicate that H2O2 is provided via the dismutation of O2-. The present results provide indirect evidence that the cytochrome c reductase activity is not a component of the NADPH-dependent H2O2 generator, since it was removed by washing the plasma membranes with 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonic acid without affecting H2O2 generation. Spectral studies with diacetyldeuteroheme-substituted horseradish peroxidase showed that the thyroid NADPH-dependent H2O2 generator does not catalyze superoxide anion formation. The O2- adduct compound (compound III) was formed but was completely inhibited by catalase, indicating that the initial product was H2O2. The rate of NADPH oxidation also increased in the presence of diacetylheme peroxidase. This increase was blocked by catalase and was greatly enhanced by superoxide dismutase. The O2- adduct compound (compound III) was produced in the presence of NADPH when glucose-glucose oxidase (which does not produce O2-) was used as the H2O2 generator. NADPH oxidation occurred simultaneously and was enhanced by superoxide dismutase. We conclude that O2- formation occurs in the presence of an H2O2 generator, diacetylheme peroxidase and NADPH, but that it is not the primary product of the H2O2 generator. We suggest that O2- formation results from oxidation of NADPH, catalyzed by the diacetylheme peroxidase compound I, producing NADP degree, which in turn reacts with O2 to give O2-.

The reduction of 2,6-dichloroindophenol (DCIP) by direct interaction with NADPH was studied. The results indicate that reduction proceeds via a direct electron transfer from NADPH to DCIP, with no oxygen consumption, and a rate constant of k = 4.69 M-1.s-1. The reduced DCIP can rapidly transfer its electrons to potassium ferricyanide (K3Fe(CN)6) or ferricytochrome c, but not to nitro blue tetrazolium. Superoxide dismutase inhibits DCIP reduction in an oxygen-dependent manner by favoring the reoxidation of the reduced DCIP. We therefore conclude DCIP is not suitable for detecting O2- when the nucleotides NADH or NADPH are present.

Phospholipids and cholesterol combine with a protein fraction (IgA and an acid polypeptide) in bile to form the bile lipoprotein complex. We wished to determine whether lysosomes participated only on IgA secretion or if their secretory role also involved the lipid components of the bile complex. This aspect was studied with a single acute injection of chloroquine, a lysosomotropic drug. The results show that a nonnegligible quantity of IgA travels through the lysosomes. In addition, phospholipid and cholesterol levels undergo a significant (P less than 0.05) decrease 1 hr after injection before increasing to normal levels. In contrast to the total inhibition of protein secretion (beta-glucuronidase, acid phosphatase), a transitory decrease of the secretion of bile lipids takes place that suggest secretory mechanisms involving organelles other than lysosomes.

Twenty-one male Wistar rats, 13 weeks old, were fed ad libitum hyperlipidic diets (28% fats) loaded with cholesterol (1.2%) for 5 weeks. One group of 11 rats was fed saturated fats (diet group "S") and another group of 10 rats was fed polyunsaturated fats (diet group "PU"). On the day they were sacrificed 10 of the rats were injected intravenously with 1 mg of filipin. Contrary to the rats in diet group "PU," the rats in diet group "S" treated with filipin presented certain characteristics that were not found in the nontreated group: They provided evidence of biliary cholestasis accompanied by a decline in the level of secretion of bile salts and phospholipids into bile. The concentrations of both free and esterified cholesterol in plasma fell and the amount of (esterified) hepatic cholesterol rose, although there was no change due to the filipin in the amounts of hepatic phospholipids. Explanatory hypotheses for these phenomena were considered, first, at the site of plasma membranes where filipin binds selectively to the cholesterol in the membrane, causing a disruption which probably disturbs the absorbance of circulating lipoproteins, especially that of hepatocyte cells, particularly in diet group "PU." Second, the effects of filipin on subcellular membranes seem to disturb the secretion of lipids and lipoproteins into bile and plasma, especially in diet group "S." Last, at the intracellular level, filipin appears to have a blocking effect on the organelles involved in biliary lipid secretion. The activity of certain enzymes such as cholesterol esterase may also be blocked, particularly in diet group "S," which would explain the accumulation of esterified cholesterol in liver.

Hog thyroid plasma membrane preparations containing a Ca2+-regulated NADPH-dependent H2O2-generating system were studied. The Ca2+-dependent reductase activities of ferricytochrome c, 2,6-dichloroindophenol, nitroblue tetrazolium, and potassium ferricyanide were tested and the effect of these scavengers on H2O2 formation, NADPH oxidation and O2 consumption were measured, with the following results. 1. Thyroid plasma membrane Ca2+-independent cytochrome c reduction was not catalyzed by the NADPH-dependent H2O2-generating system. This activity was superoxide-dismutase-insensitive. 2. Of the three other electron scavengers tested, only K3Fe(CN)6 was clearly, but partially reduced in a Ca2+-dependent manner. 3. Though the NADPH-dependent reduction of nitroblue tetrazolium was very low and superoxide-dismutase-insensitive, nitroblue tetrazolium inhibited O2 consumption, H2O2 formation and NADPH oxidation, indicating that nitroblue tetrazolium inhibits the H2O2-generating system. We conclude that the thyroid plasma membrane H2O2-generating system does not or liberate O2- and that Ca2+ controls the first step (NADPH oxidation) of the H2O2-generating system.

Studies of the serological development after 26 renal transplants confirm the high frequency of antibody rises not only against the herpes virus group, but also against other virus groups such as measles, Coxsackie B viruses. These antibody rises correlate with febrile episodes and hepatic dysfunction in which CMV is the most often involved. However, the frequency of antibody rises against various viral antigens without any clinical event to suggest viral etiology; the lack of concomitant virus isolation (except the herpes group), as well as the ocurrence of simultaneous antibody rises against several viruses, all suggest that some of these various antibody rises observed may be related to immunological dysfunction rather than to virus infection.

A survey of 4,452 pregnant women taken to find hepatitis B surface antigen revealed 28 asymptomatic chronic carriers. At birth, 16 of 17 infants studied were negative for HBsAg, whereas anti-HBc was present in all patients at a titer similar to that of the mother. Twelve children were followed for 6 to 18 months. In four of them, HBsAg remained negative and anti-HBc titers progressively decreased and were undetectable when tested after the age of 6 months. In eight infants, HBsAg became positive after an average time of 48 days. Elimination of HBsAg occurred in seven infants; five of them had clinical and biological manifestations of mild hepatitis. Sequential determinations of total complement and C components in three patients of the latter group showed dperession of complement at the time of appearance of clinical manifestations. In the patient who became an HBsAg carrier as well as in three infants who remained HBsAg negative, no decrease in complement titers was observed. These results indicate that vertical transmission from carrier mothers can occur in a low prevalence area and that neonatally infected children are capable of active elimination of HBV.

We have isolated substances of molecular weight ranging between 350 and 2,000 daltons from ultrafiltrates of 3 patients treated by maintenance haemodialysis for chronic renal failure. Such substances might have a role in the genesis of uraemic toxicity. We have chiefly studied their carbohydrate content. Material was fractionated according to a procedure previously used to urine in healthy controls. Consecutive ion exchange, charcoal Celite and paper chromatography lead to the isolation and purification of oligosaccharides, glycopeptides, glucuronoconjugates and peptides. The different classes of carbohydrate material present in dialysis fluids from uraemic patients are close to those formed in normal urines. All the oligosaccharides in renal failure urine had have identified in normal urine. In a previous studies we have demonstrated that the levels of glucuronoconjugates are higher in the blood of uraemic patients. The glucuronoconjugates and their aglycones could have a toxic effect but a great part of them is removed by hemodialysis.

Solubilization of the thyroid particulate-associated NADPH-dependent H2O2 generating system has been tested with different detergents; (3-(3-cholamidopropyl)-dimethylammonio)1-propane sulfonate (CHAPS) was found to be the best of the six detergents tested. The ratio of H2O2 generation to NADPH oxidation was similar for CHAPS extract and native particulate material. CHAPS was also the only detergent able to preserve the Ca++-sensitivity of the NADPH oxidase. Solubilization of this enzyme allowed the determination of some of its characteristics: specificity for divalent cations, apparent Km for NADPH, optimum pH and sensitivity to SH- reagents.

Differences in mouse hepatitis virus 3 (MHV3) sensitivity among mouse strains are mainly determined by H-2-related and -nonrelated genetic factors. Reciprocal chimerism was therefore established between two H-2a compatible pairs of strains that differ widely in their susceptibility to MHV3: a) A/J and B10.A, respectively resistant and highly susceptible; b) A/J and A/Sn, respectively resistant and semisusceptible. Chimeric mice were challenged with 100 LD50 of MHV3, 30 or 90 days after X-irradiation (900 R) and bone marrow reconstitution. Results showed that sensitivity of recipients was similar either to that of the recipient strain or to that of the donor strain when chimeric mice were tested 30 or 90 days, respectively, after reconstitution. In addition, no paralysis occurred in surviving animals. These data indicate, therefore, that resistance or susceptibility to MHV3 is expressed intrinsically in some population(s) of hematopoietic-derived cells, which is radioresistant and has a life span of more than 30 days and less than 90 days. Additional experiments showed that X-irradiated A/J recipients reconstituted with A/J bone-marrow cells were protected against MHV3 challenge with spleen cells, with a mixture of spleen cell populations or of adherent spleen cells and thymocytes originating from A/J donors. Transfer of protection to recipients by using similar cell populations provided by semisusceptible A/Sn donors required the administration of five times more cells. Results suggest that two complementary mechanisms are required to confer resistance to MHV3: a) a gene(s) for resistance that may operate at the level of macrophages, and b) cells capable of mounting an efficient immune response. The reduced efficiency of A/Sn spleen cells suggests that semisusceptibility to MHV3 may be related to partial quantitative or functional immune defect.

The regulating process of cholesterol in the liver was studied in relation to its exogenous contribution in the rats fed high-fat (28%) high-cholesterol (1.2%) diets rich in saturated (S) fat (lard) or polyunsaturated (PU) fat (corn oil). Accordingly, the fate of 14C free cholesterol originating from high- or low-density lipoproteins (LDL) was examined in the biliary, hepatic and plasmatic lipids, as well as the activity of two key enzymes in the metabolism of lipoproteins: lipoprotein lipase (LPL) and lecithin cholesterol acyl transferase (LCAT). The LPL activity increased in the S diet, in comparison to the PU diet or to a low fat (6%) control (C) diet and the LCAT activity increased but not significantly in the PU diet. In bile the secretion of 14C-cholesterol and 14C-bile salts originating from 14C-cholesterol-HDL increased in the S diet compared to the PU diet and a C diet [previous results]. S and PU diets increased to the same extent the hepatic storage of 14C-esterified cholesterol originating from LDL, compared to the C diet. This cholesterol would contribute to a greater extent to the hepatic synthesis of the lipoproteins destined for the plasma in the case of the S diet than that of PU diet. These results may be explained by the adaptation of hepatic acyl cholesterol acyl transferase and cholesterolesterase to both high-fat-diet enzymes acting simultaneously on the two free and esterified cholesterol compartments. It resulted in an important redistribution of the cholesterol of these two compartments between plasma, bile and liver.

Complement (C) and C components were studied in 18 children with severe viral hepatitis. Results were compared to similar determinations performed in 8 patients with toxic hepatitis related to the ingestion of Amanita Phalloïdes. Hemolytic activities and serum concentrations of C and C components were markedly decreased (less than 2 SD) in both groups of patients. Sequential determinations of C components showed different patterns according to the outcome: either a rapid return to normal in patients who healed or persistence of low levels in patients who died. Such a pattern was similar to that of the prothrombin time. In patients with viral hepatitis a rapid disappearance of C4 and C3 was observed following fresh plasma or fresh blood transfusion. In addition, C4 cleavage was shown in plasma of 8/8 patients. These data suggest that the complement depression observed in fulminant viral hepatitis is related to both consumption and deficient synthesis.

A thyroid particulate fraction contains an NADPH-dependent H2O2-generating enzyme which requires Ca2+ for activity. A Chaps solubilized extract of the thyroid particulate fraction partially purified by DEAE chromatography did not show a dependence on Ca2+ for activity. Preincubation of the particulate fraction with Ca2+ yielded a preparation insensitive to Ca2+. The non-particulate fraction obtained after incubation of the particles in the presence of Ca2+ was able to inhibit, in the presence of EGTA, the Ca2+-desensitized particulate fraction and the enzyme isolated on DEAE. It is concluded that the reversible Ca2+ activation of the NADPH-dependent H2O2 generation was modulated in porcine thyroid tissue by (a) calcium-releasable inhibitor protein(s).

In order to study the relationship between bile cholesterol and free cholesterol carried by high and low density lipoproteins (HDL and LDL), 10 male Wistar rats, 11 weeks old and fed with a standard diet were divided into 3 groups which received an intravenous infusion (jugular vein) of either LDL, HDL or liposomes. Liposomes were used for comparison because they are assimilated by hepatocytes, but are not recognized by specific receptors. HDL isolated from rat sera were labeled with [14C]cholesterol by molecular exchange and LDL were labeled by exchange with [14C]cholesterol incorporated into phosphatidyl choline/cholesterol liposomes. The peaks of radioactivity appeared in bile 30 min after the HDL or liposome injection and after 210 min for the LDL injection. The kinetic behavior of the cholesterol carried by the liposomes was quite similar to that of cholesterol carried by HDL. Cholesterol carried by HDL was metabolized in bile salts faster than that carried by LDL: cholesterol-HDL or cholesterol-liposomes contributed to the same extent to the secretion of bile cholesterol (15 and 11%, respectively, of the injected dose), LDL (20% of the injected dose). However, the main part of [14C]cholesterol from HDL, LDL or liposomes was metabolized in bile salts. Thus, cholesterol from an exogenous source seemed to be used mainly as a substrate for bile salts. Our study revealed a difference between the hepatic metabolism of HDL, liposomes and LDL in the rat: the kinetic difference between the secretions of the radioactive compounds in bile may be explained by differences in assimilation, intracellular pathways or bile secretion.