pNiXa, a Ni(2+)-binding protein in Xenopus oocytes and embryos, shows identity to Ep45, an estrogen-regulated hepatic serpin.

A Ni(2+)-binding protein (pNiXa, 45 kD, pI 8.5) discovered in Xenopus embryos, was isolated from oocytes. Based on amino acid sequences, pNiXa belongs to the serpin superfamily and shows identity to the cDNA sequence of Ep45, an estrogen-regulated hepatic serpin that contains an (HX)n-motif found in eukaryotic transcription factors. Nondenatured pNiXa, purified by Ni-affinity chromatography, inhibited bovine alpha-chymotrypsin. The presence of pNiXa in embryos when they are susceptible to Ni2+, the high avidity of pNiXa for Ni2+, and the (HX)n-motif point to pNiXa as a molecular target of Ni(2+)-teratogenesis.

[New pathogenetic aspects of the relations between viral and microbial infections and peripheral vasculopathies]. / Quelques nouveaux aspects pathogéniques des relations entre certaines infections à virus et inframicrobiennes et des vasculopathies périphériques.

The immunofluorescence technique using antisera against some viruses and inframicrobes allowed the detection of pathogens in altered vascular tissues (arteritis and phlebitis-phlebectasia). Pathomorphological aspects and some dehydrogenase activities in these patients were also investigated.

Toxicity to alveolar macrophages in rats following parenteral injection of nickel chloride.

Alveolar macrophages collected by pulmonary lavage from male Fischer-344 rats at intervals (1-72 hr) after NiCl2 injection (62-500 mumol/kg, sc) were tested by several techniques. Within 1 to 4 hr, the macrophages showed morphological and biochemical signs of activation (hypertrophy, ruffled plasma membrane, increased cyclic AMP concentration, and markedly diminished 5'-nucleotidase activity, assayed by concanavalin A inhibition). Functional impairment (reduced phagocytic activity) was first seen at 24 hr; lipid peroxidation (increased malondialdehyde concentration) was not detected until 48 hr. Dose- and time-related effects of NiCl2 on 5'-nucleotidase activity, phagocytic activity, malondialdehyde concentration, and nickel content of alveolar macrophages were observed 24 to 72 hr postinjection. Diminished cell viability occurred only at 72 hr after the highest dosage of NiCl2. In alveolar macrophages from 63NiCl2-treated rats, 63Ni was located primarily in the cytoplasm, based upon liquid scintillation counting and autoradiography; fractionations of macrophage cytosol by gel filtration chromatography showed that 63Ni was bound to several high- and low-molecular-weight constituents. This study demonstrates that sc administration of NiCl2 to rats caused nickel uptake into and activation of alveolar macrophages, followed by reduced phagocytic capacity. The alveolar macrophage was a cellular target for nickel toxicity following parenteral exposure to NiCl2.

Hepatic lipid peroxidation in CoCl2-treated rats, evidenced by elevated concentrations of thiobarbituric acid chromogens.

Parenteral administration of cobalt chloride (CoCl2) to male Fischer-344 rats caused hepatic lipid peroxidation, as evidenced by the thiobarbituric acid (TBA) reaction for malondialdehyde (MDA) and related chromogens in fresh liver homogenates. After a single sc injection of CoCl2 (300 mumol/kg, body wt.), hepatic concentrations of TBA-chromogens became significantly increased by 4 h and reached peak levels at 24-72 h post-injection. For example, in rats killed 48 h post-injection, hepatic TBA-chromogens (mean +/- SD, N = 7) averaged 607 +/- 141 nmol/g of tissue, wet wt., (P less than 0.01 versus 245 +/- 73 nmol/g in 19 controls). A dose-effect relationship was observed between CoCl2 dosages (100 to 450 mumol/kg, body wt.) and hepatic concentrations of TBA-chromogens in rats killed 24 h post-injection. This study indicates that hepatic lipid peroxidation develops as a consequence of acute CoCl2 toxicity in rats.

Acute thymic involution and increased lipoperoxides in thymus of nickel chloride-treated rats.

Acute thymic involution occurred in male, Fischer-344 rats following a single injection of nickel chloride (0.5 mmol/kg, sc). In nickel-treated rats, the mean thymic weight became significantly decreased at 24 h, continued to diminish at 48 h, and reached 24% of the control value at 72 h post-injection. The ratio of thymic weight to body weight (mg/g) decreased from 1.24 +/- 0.05 (SD) in control rats to 0.30 +/- 0.05 in nickel-treated rats at 72 h post-injection (p less than 0.01). Concentrations of thymic lipoperoxides were assayed by the thiobarbituric acid reaction, with HPLC separation and spectrophotometric quantitation of the malondialdehyde-thiobarbituric acid complex. The mean concentration of thymic lipoperoxides was unchanged at 24 h, increased 2-fold at 48 h, and reached 8-times the control value at 72 h after nickel chloride injection. On histological examination, sections of thymus from nickel-treated rats showed moderate to profound depletion of cortical lymphocytes at 72 h post-injection. Marked degenerative changes were noted in cortical lymphocytes, with pyknosis and karyorrhexis; swelling and vacuolation were evident in thymic reticular epithelial cells. The time-courses of the biochemical and histopathological responses suggest that the lipid peroxidation may be an end-result, rather than a cause, of thymic involution and injury to thymic lymphocytes in nickel-treated rats.

Lipoperoxides in plasma as measured by liquid-chromatographic separation of malondialdehyde-thiobarbituric acid adduct.

This assay of plasma lipoperoxides involves hydrolysis in dilute H3PO4 at 100 degrees C; complexation of malondialdehyde (MDA), a hydrolysis product, with thiobarbituric acid (TBA); methanol precipitation of plasma proteins; fractionation of the protein-free extract on a C18 column; and spectrophotometric quantification of the MDA-TBA adduct at 532 nm. The detection limit was 0.15 mumol of MDA per liter of plasma. Run-to-run precision (CV) averaged 8 to 13%. Analytical recovery of MDA after addition of tetraethoxypropane standards to 21 specimens of human or rat plasma averaged 98% (SD 7%). Lipoperoxide concentrations (as MDA) averaged 0.60 (SD 0.13) mumol/L in plasma specimens from 41 healthy persons and 1.4 (SD 0.3) mumol/L in plasma specimens from 12 control rats. Mean lipoperoxide concentrations were 1.5 to 2.3 times as great in plasma sampled from rats one to three days after subcutaneous administration of NiCl2 at dosages (250 to 750 mumol per kilogram body wt) previously shown to induce lipid peroxidation in lung, liver, and kidney.

[Possible relation between viruses and oromaxillofacial tumors. I. Demonstration of herpes antigens and anti-herpes antibodies]. / Relations possibles entres virus et tumeurs bucco-maxillo-faciales. Note I. Mise en évidence d'antigènes herpétiques et d'anticorps anti-herpétiques.

Herpes virus type 1 and 2 antigens were detected in cells originating from malignant tumors with oromaxillofacial localization. A higher incidence of type 1 antigens was noted, as compared to the type 2. Investigated antigens were detected more frequently in the tumors of ectodermal origin. Antiherpesvirus FC antibodies were found in 74.5% of the tumor bearing subjects, but only in 22.3% of the controls.

[Possible relation between viruses and oromaxillofacial tumors. IV. Presence of the herpes antigen and anti-herpes antibodies in patients with tumors of the parotid gland]. / Relations possibles entre virus et tumeurs bucco-maxillo-faciales. Note IV. Présence de l'antigène herpétique et des anticorps anti-herpétiques chez des malades porteurs de tumeurs de la glande parotide.

The presence of herpesvirus antigen and of antiherpesvirus antibodies was detected in 46% and in 36.3%, respectively, of the patients bearing parotid gland tumors. Very high titers of antiherpesvirus antibodies were found in 70.6% of the patients with nasopharyngeal carcinoma.

[Possible relation between viruses and oromaxillofacial tumors. III. Demonstration of the SV40 antigen and anti-SV40 antibodies in patients with tumors of the parotid gland]. / Relations possibles entre virus et tumeurs bucco-maxillo-faciales. Note III. Mise en évidence de l'antigène SV40 et des anticorps anti-SV40 chez des malades ayant des tumeurs de la glande parotide.

SV40 antigen was detected in 7 of 13 malignant tumors developed in the head and neck region. Specific complement fixing antibodies were found in all the patients with the SV40 antigen present in the parotid gland tumoral cells. Incidence of the anti-SV40 complement fixing antibodies in parotid gland tumor bearing patients was of 69.6%.

[Possible relation between viruses and oromaxillofacial tumors. II. Research on the presence of the SV40 antigen and specific antibodies in patients with oromaxillofacial tumors]. / Relations possibles entre virus et tumeurs bucco-maxillo-faciales. Note II. Recherches sur la présence de l'antigène SV40 et des anticorps spécifiques chez des malades porteurs de tumeurs bucco-maxillo-faciales.

SV40 antigen was detected on sections through malignant oromaxillofacial tumors in 47.8% of the cases, with a higher incidence among the epithelium originating tumors. Anti-SV40 CF antibodies were detected in 83.6% of the examined subjects with cancer, but only in 57.9% of the blood donors. The titers were quite low in general. The results are discussed from the point of view of the immunologic status of the patients.

[Possible relation between viruses and oromaxillofacial tumors. V. Demonstration of hemagglutination-inhibiting anti-BK virus antibodies in patients with tumors of the parotid gland]. / Relations possibles entre virus et tumeurs bucco-maxillo-faciales. Note V. Mise en évidence des anticorps inhibiteurs de l'hémagglutination anti-virus BK chez des malades porteurs de tumeurs de la glande parotide.

Anti-BK-virus hemagglutination inhibiting antibodies were revealed in 81.8% of the patients with parotid gland tumors. Results of the investigations conducted on oromaxillofacial tumors including the parotid gland ones are discussed from the point of view of the presence of viral antigens (herpes-, SV40 and BK-viruses) and of specific antibodies. Possible implication of the papova viruses in the etiopathogenesis of the parotid gland tumors in humans are also discussed.

Hepatic toxicity of nickel chloride in rats.

Enhanced lipid peroxidation was observed in livers of rats killed 24 hr after sc injection of nickel chloride (NiCl2) (750 mumol per kg), as evidenced by 13-fold increase of conjugated dienes in microsomal lipids and 4-fold increase of thiobarbituric acid (TBA) chromogens in hepatic cytosol. Histologic examination of livers from rats killed one to three days after NiCl2 injection (500 mumol per kg) showed microvesicular fatty metamorphosis, mild hydropic degeneration, and foci of inflammation. Microvesicular steatosis of hepatocytes was confirmed by electron microscopy. Dose-related increases of serum aspartate aminotransferase (ALT) activity (up to 7-fold vs controls) and alanine aminotransferase (ALT) activity (up to 3-fold vs controls) were observed 24 hr after injection of NiCl2 (125 to 750 mumol per kg); diminished serum alkaline phosphatase activity (up to 72 percent reduction vs controls) was seen at NiCl2 dosages from 375 to 750 mumol per kg. Diethyldithiocarbamate did not influence the effects of NiCl2 on TBA-chromogens in liver homogenates or on serum AST and ALT activities but acted synergistically with NiCl2 to diminish serum alkaline phosphatase activity and to increase serum bilirubin concentration. This study demonstrates that parenteral administration of NiCl2 to rats produces acute hepatic toxicity, with enhanced lipid peroxidation, microvesicular steatosis, and increased serum AST and ALT activities.

Increased lipid peroxidation in tissues of nickel chloride-treated rats.

Parenteral administration of nickel chloride (NiCl2) to rats enhanced lipid peroxidation in liver, kidney, and lung (but not in brain, heart, spleen, or testis), as measured by the thiobarbituric acid reaction for malondialdehyde (MDA) and related chromogens in fresh tissue homogenates. After sc injection of NiCl2 (0.75 mmol per kg body wt), MDA concentrations in liver and kidney became significantly increased by nine h and reached peak values at 48 h. For example, in nine rats killed 48 h after the NiCl2 injection, hepatic MDA concentrations averaged 2.5 +/- 1.0 mumol per g dry wt (P less than 0.001 versus 0.5 +/- 0.3 mumol per g in 30 controls). Dose-effect relationships for lipid peroxidation in liver and kidney were observed with NiCl2 dosages ranging from 0.12 to 0.75 mmol per kg, sc. Intrarenal administration of a carcinogenic nickel compound, nickel subsulfide (Ni3S2, 0.36 mmol per kg body wt), did not affect MDA concentrations in the injected kidneys of rats killed one to 20 days post-injection. The results of this study implicate lipid peroxidation as a molecular mechanism for cell injury in acute NiCl2 poisoning, but they do not furnish any evidence that lipid peroxidation is involved in the initiation of nickel carcinogenesis.

Effects of diethyldithiocarbamate and nickel chloride on glutathione and trace metal concentrations in rat liver.

Concentrations of reduced glutathione (GSH) and oxidized glutathione (GSSG) and 4 trace metals (Ni, Cu, Mn, Zn) were measured in livers from rats treated with sodium diethyldithiocarbamate (DDC, 0.67 or 1.33 mmol/kg, i.m.) and NiCl2 (0.25 or 0.50 mmol/kg, s.c.), singly or in combination. In rats treated with DDC or NiCl2, singly, hepatic GSH was diminished at 4 h and returned to control levels (or slightly above) at 17 h. In rats that received DDC plus NiCl2, hepatic GSH was not diminished at 4 h after increased 1.4-1.8-fold at 17 h. Hepatic GSSG was diminished at 4 h after NiCl2 treatment and returned to control values at 17 h; hepatic GSSG did not differ from control values at 4 h or 17 h after treatment with DDC, alone or combined with NiCl2. Hepatic Ni was below the detection limit (approximately 20 nmol/g) in control and DDC-treated rats; hepatic Ni was increased to 53 +/- 26 (S.D.) nmol/g at 17 h after treatment with NiCl2 alone, and was increased 6-fold (308 +/- 63 nmol/g) in rats that received Ni plus DDC. Under the same conditions, hepatic Zn was increased 33% or 41%, respectively, in rats that received NiCl2 or DDC, singly, and was not further increased by combined treatment; hepatic Cu and Mn concentrations were unaffected by NiCl2 or DDC, singly, but were diminished in rats that received NiCl2 and DDC. This study suggests: (a) that increased hepatic uptake of Ni is largely responsible for the synergistic induction of heme oxygenase activity in rats treated with NiCl2 and DDC; and (b) that increased hepatic uptake of Zn contributes to the induction of hepatic metallothionein by NiCl2 and DDC.