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Isolated albumin almost always contains polymerized forms which appear during preparation and storage of the protein. The proportion of polymerized forms reflects the degree of stability of the solution. The quantitative estimation of the polymers is usually performed by gel chromatography. In this work, the high resolution power of polyacrylamide gradient gel electrophoresis (Gradient PAGE) was used to separate the polymers present in the preparations of human serum albumin. The analysis of the different peaks obtained by gel chromatography allows to conclude that peak 1 contains aggregates and high polymers, peak 2 trimer and dimer and peak 3 the monomer of albumin. The aggregates of the peak 1 can be dissociated by SDS and correspond, in gradient PAGE, to the high polymers. By using gradient PAGE in the presence of SDS under the conditions described in this paper, it is possible to estimate the proportion of high polymers and aggregates present in albumin preparations. These results are similar to those obtained by chromatography followed by a protein assay but noticeably inferior to those resulting from measurements performed by absorbance at 280 nm.

Airway inflammation due to exposure to a wide variety of agents encountered in the workplace is a major cause of occupational asthma. This article reviews major examples of occupational asthma linked to airway inflammation, including their epidemiology, pathophysiology, and clinical course.

The relatives of a 82 years old female patients with a Waldenstrom Macroglobulinemia were submitted to clinical and hematological investigations. A diclonal gammapathy (IgG kappa and IgG lambda) was found in the sister's serum and was related to a myeloma. A noticiable Bence Jones Proteinuria (light chain of lambda type) was equally found in serum and urines of the propositus brother but without any clinical or hematological evidence of myeloma. The examination of two other sisters of these patients does not reveal any immunological of hematological disorder. This new observation underlines the interest of a systematical investigation in the siblings of the patients with monoclonal gammapathies.

Low density lipoprotein (LDL + VLDL) concentrations, were measured in 48 patients with multiple myelomatosis, or Waldenström's macroglobulinaemia (malignant monoclonal gammopathies) and in 42 patients with "asymptomatic" benign monoclonal gammopathies (M.G.). In patients with malignant M.G., the level of LDL plus VLDL was significantly lower than in patients with "asymptomatic" M.G. who exhibit a normal level.

Simultaneous occurrence of "myasthenia gravis" and monoclonal "asymptomatic" gammapathy has been observed in a 83 years old patient. Antistriated muscle antibodies could be detected at a level of 1/1,000 but no antiacetylcholine receptors antibodies. The antibody activity is located both in the monoclonal IgG kappa and in the polyclonal IgG. This case illustrates the frequent occurrence of several auto-immune disorders in a same patient.

Inter-alpha-trypsin inhibitor (I alpha I) has been purified from C.N.T.S. fraction III as starting material. The purification procedure includes D.E.A.E. cellulose chromatography and gel filtration on G 150 Sephadex in the presence of EDTA. The purified protein gives one precipitation line in immunoelectrophoresis against anti-whole human sérum. It reacts only with an anti I alpha I immune serum and possesses a strong antitryptic activity. When studied in starch or polyacrylamide gel electrophoresis 2 components are observed, each of them having the same antigenic structure and the same antitryptic activity as the crude preparation. The slower and less important component is dissociated by 0,1% SDS. The molecular weight estimation of I alpha I BY PAA/SDS is about 180,000. This result is not modified by the presence of 1% beta mercaptoethanol indicating that I alpha I consists of one polypeptide chain. Crude preparation reveals under the same electrophoretical conditions small amounts of low molecular weight components (135,000 52,000 and 26,000) which can be due to a proteolytic action on I alpha I. Indeed plasmin is able to produce such fragments having an antitryptic activity as shown by fibrin/polyacrylamide gel electrophoresis. The relationship between small molecular weight inhibitors of human serum and bronchial secretions and those obtained after degradation of I alpha I by plasmin is discussed.

A human monoclonal IgG anti-Rhesus(D) (H2D5D2) obtained after transformation of lymphoid cells by Epstein-Barr virus was purified from culture supernatant by affinity chromatography. Rabbits were immunized with the monoclonal anti-D. The rabbit antisera, after appropriate absorption, reacted only against the immunizing monoclonal anti-D. The antiidiotypic antibodies (anti-id ab) were purified and gave a complete inhibition of the monoclonal anti-D, whereas no inhibition was observed with monoclonal or polyclonal IgG or with sera containing high titres of anti-D. Polyclonal anti-D obtained from 118 immunized blood donors were coated on Rh-positive red cells. Agglutination by anti-id ab was observed in 4 cases (3.4%), indicating a cross-reactivity between the monoclonal anti-D and the polyclonal anti-D present in the sera of some immunized donors.

Circulating proalbumin in humans has been described in two distinct events: genetic variants of proalbumin related to mutations at the cleavage site, and normal proalbumin related to an abnormal cleaving enzyme system. We report a case of acquired proalbuminemia that appeared after an acute episode of hepatitis related to the delta agent, in a chronic carrier of hepatitis B virus. This component, not present in normal plasma, was identified as proalbumin by immunological methods. It was indistinguishable from the molecule normally present in hepatocytes as judged by electrophoretic mobility, limited susceptibility to tryptic digestion, and its inability to bind labeled Ni. We suggest that this release of proalbumin is related to the concurrent presence of both hepatitis B and delta virus in some of the infected hepatocytes.

The airway edema that develops in guinea pigs after exposure to toluene diisocyanate (TDI) requires the presence of polymorphonuclear leukocytes (PMN). To determine whether this airway edema is mediated by the release of hydrogen peroxide from PMN, we treated animals intravenously with catalase bound to polyethylene glycol and examined the extravasation of Evans blue dye into the tracheal wall after exposure to air or 3 ppm TDI for 1 h. Catalase (25,000, 100,000, and 300,000 IU/kg) caused a dose-dependent inhibition of the TDI-induced increase in dye extravasation. However, treatment with catalase, inactivated at the peroxide binding site with 3-aminotriazole, inhibited dye extravasation after exposure to TDI as effectively as the equimolar 100,000 IU/kg dose of active catalase. Injection of polyethylene glycol alone was without effect. Dose-dependent decreases in extravascular migration of PMN and in circulating PMN also were noted after catalase treatment. These results suggest that the catalase preparations used in these studies inhibited the PMN-dependent airway edema by an effect other than hydrogen peroxide scavenging. Examination of this and other commercially available catalase preparations revealed trace concentrations of endotoxin at levels that could be responsible for the observed effects on PMN function. Treatment of animals with doses of Escherichia coli endotoxin similar to those inadvertantly administered to the catalase-treated groups (0.1 ng/kg to 100 ng/kg, intravenously) inhibited TDI-induced extravasation of Evans blue dye in a dose-dependent manner. These results suggest that contaminating endotoxin may contribute to some of the protective effects of preparations of catalase observed in previous studies of vascular permeability.

An 85-year-old woman without personal history of haemorrhages was found to have qualitative and quantitative deficiency of Factor VIII persisting at least 6 months. Asymptomatic monoclonal IgG kappa gammopathy was also discovered in the same patient, together with a circulating inhibitor of ristocetin co-factor. The fact that the inhibitory effect was reduced after the patient's serum IgG's were bound to staphylococcal protein A suggests that the inhibitor belonged to that category of immunoglobulins, although the authors were unable to detect it after elution.

In order to clarify the actual nomenclature of the albumin allotypes in French and Italian populations we compared the samples collected in our laboratory to those kindly supplied by Dr. Porta (Italy) with reference to albumin variants classified in starch gel electrophoresis using three buffer systems by Weitkamp. The inherited human albumin variants can be classified on the basis of their relative mobilities on cellulose acetate electrophoresis compatively to human transferrin mobility. The relative mobility of each variant can be expressed by the following ratio: migration distance of the variant versus migration distance of the normal albumin where zero represents the transferrin mobility. Using three buffers system at pH 8.6, 5.0 and 6.9, it is possible to distinguish some albumin variants having a same mobility at alkaline pH and different mobilities at acidic pH. In the european area, eleven albumin variants are distinguishable on the basis of their relative mobilities at different pH: four Fast moving variants: Gent, Vanves (a new variant described here), Reading and CN/BL, and seven Slow moving variants: MI/MI Slow, GE/CT, SO/BS (or D type), Pollibauer, Gainesville, Roma and B type. Thirty-six sera from unrelated subjects with genetic bisalbuminemia were analyzed in our laboratory. Their distribution was as follows: B type (22), Pollibauer (9), SO/BS (2), Gainesville (1), Gent (1) and Vanves type (1). The frequency of bisalbuminemia was 0.35 per 1,000 in a population of 19,949 blood donors.

Pure alpha2M is prepared with fresh plasma as starting material, to prevent the interaction of alpha2M from proteolytic enzymes of plasma such as thrombin, plasmin and kallikrein. During the purification steps, polybrene and aprotin are used as inhibitors and plasminogen is absorbed onto bentonite. When alpha 2M is submitted to polyacrylamide gel electrophoresis (PAA) containing 0.1% SDS, a complete dissociation in two half-molecules of MW 380,000 occurs. When alpha2M is incubated in 1% SDS and 1% beta-mercaptoethanol as reducing agent, only one component of MW 190,000 is observed in PAA-SDS. This experiments show that the alpha2M molecule consist of two symetric halves of same MW (380,000) linked by non covalent bonds. Each two-half-molecules is made of two polypeptides chains MW 190,000 linked by disulfide bonds. Thus alpha2M molecule contains four polypeptides chains having a same MW. The same techniques were applied to the study of alaph2M proteinases complexes. Three different proteinases (plasmin, trypsin and papain) were used in these experiments. Trypsin and papain are commercialy available. Plasminogen was obtained by affinity chromatography and activated into plasmin by insoluble streptokinase fixed on PAB cellulose.

Sixty-one human myeloma proteins (HMP) from patients with multiple myeloma and Waldenström macroglobulinemia were tested for anti-idiotypic (Id) activity against autoantibodies to double-stranded DNA, small nuclear ribonucleoproteins, and human thyroglobulin (HTg), by competitive radioimmunoassays and enzyme immunoassays. An IgG1, lambda HMP from patient BEN with anti-Id activity against antibodies to HTg is reported. IgG1 BEN was not directed toward human Fc fragments and its activity was not related to allotypic determinants. IgG1 BEN molecules recognized Id determinants (idiotopes) on F(ab')2 anti-HTg fragments, but not idiotopes of F(ab')2 fragments of antibodies of other specificities. This observation supports the general significance of Id network interactions in regulation and diversification of immune responses in man.

We report two cases of mu-heavy-chain disease. Both patients were affected with a lymphoproliferative disease that shared several suggestive features with the previously reported cases of mu-chain disease: the presence of vacuolated plasma cells in bone marrow, a small amount of alpha 2 moving abnormal mu-chain protein, and urinary kappa Bence Jones protein in one case.

Acquired Von Willebrand syndrome is reported in four patients with monoclonal lgG : benign gammapathy in three cases, multiple myeloma in one case ; to our knowledge, this last association has not been previously reported.Coagulation abnormalities included a borderline bleeding time, a low platelet retention on glass beads, decreased levels of factor VIII coagulant activity (VIII: c), factor VIII related-antigen (VIII R: Ag) and ristocetin-induced agglutination cofactor (VIII R: Cof).The late clinical onset, the negative family history and the immunological abnormality suggest an acquired Von Willebrand syndrome. After cryoprecipitate infusion the patients did not show the expected rise and there was no secondary increment in factor VIII: c.Time-dependent inhibition of factor VIII R: Cof was found in one case only and was associated with qualitative abnormality of factor VIII R: Ag demonstrated by crossed-immunoelectrophoresis. It was not possible to interpretate this last test in the other cases, due to the very low level of factor VIII R: Ag. The inhibitor activity was neither found in the IgG fraction nor in the monoclonal IgG but disappeared after IgG adsorption on staphylococcal protein A.The factor VIII abnormalities might be related to the binding and/or destruction of factor VIII by a circulating antibody, or to the adsorption of this factor on the malignant lymphocytes.

Weitkamp et al. (1973) using starch gel electrophoresis in three different systems of buffers (pH 5, 5.6 and 6.9) distinguish at least 24 genetically determined variants. In the European area only seven variants have been observed. The study of 24 cases of bisalbuminemia observed in the french population by acetate electrophoresis at pH 8.6 and polyacrylamide agarose gel electrophoresis at pH 8.7 and pH 6.9 can distinguish three slower variants than the normal albumin. The most frequent variant is the B type observed in 16 cases. The other types are the Pollibauer type observed in 7 cases, and the Gainsville type in one case only. The incidence of these variants can be estimated to 0.7 p. 1000 individuals.

Monoclonal gammopathies (M.G.) are a group of disorders characterized by the proliferation of a single clone of plasma-cells that produce a Monoclonal Immunoglobulin (M-Ig). The presence of M-Ig can be demonstrated by electrophoresis in the patient' serum and identified by immunoelectrophoresis as a molecule containing two heavy chains of a single class and two light chains of a single type. In M.G. of malignant origin, free light chains (Bence Jones Proteins) can be evidenced in the concentrated urines. The fact that M.G. may be more frequent in certain families show the existence of a familial predisposition of this disease whereas its origin is still unknown. M.G. are often associated with malignant proliferation of B lymphocytes such as multiple myeloma, Waldenström's macroglobulinemia, heavy chain disease as well as some other lymphoproliferative disorders. However, in a certain number of cases, the malignant origin of the M.G. was not proved, because M-Ig can occur in the serum of people apparently in good health and without clinical or hematological features (asymptomatic "benign" M.G.). Asymptomatic benign M.G. have been detected in increasing numbers during the last decade due to use of cellulose acetate electrophoresis for the routine examination of patients or in the course of systemic screening in normal populations such as blood donors. At the present time, a malignant origin of M.G. cannot be proved in more than thirty per cent of case and these "asymptomatic M.G." must be follow-up by a yearly clinical, hematological and electrophoretical check-up in order to detect a possible malignant evolution. In other cases, M.G. can be associated with neoplasms of cells types not known to produce M-Ig, in cold chronic agglutinin disease, and during the course of some auto-immune disorders.

The atherogenic low density lipoproteins were evaluated using two turbidimetric methods: one based on the precipitation of LDL and VLDL by heparin-CaCl2 and the other on the precipitation of the VLDL by sodium dodecyl sulfate. In search of hyperlipoproteinemia, both tests were applied to a population of blood donors aged from 18 to 60 years. when both tests were negative, hyperlipoproteinemia can be excluded without quantitative evaluation of cholesterol and triglycerides levels. If the heparin-CaCl2 test is high and the SDS normal it can be concluded that it is a case of type IIa hyperlipidemia. When both test are high, further investigations are required. In the present study, corresponding to the screening of the sera from 2656 blood donors (including 1280 males and 1376 females), the high level of atherogenic lipoproteins is most frequently observed in the males (10 per cent for men and 2,5 per cent for women). This frequency increases with age.

Using agarose gel coated on GelBond film sheets, and using Coomassie blue stain followed by silver stain, a sensitive microassay has been developed for detecting small amounts of antigen-antibody precipitates, and for quantitating low concentrations of antigen. In order to obtain a high sensitivity, antigen-antibody ratios were adjusted imperatively close to the equivalence in double-diffusion, and for quantitative estimation, single radial immunodiffusion was performed, according to Mancini, by measuring circles at the end point. The use of a double staining procedure allows to detect as little as 8 micrograms/ml antigen by Ouchterlony and 200 ng/ml by Mancini technique. The sensitivity of the method is 100 times greater than classical techniques and other advantages such as the need for minimal amounts of unconcentrated samples, the absence of radioactive labelling, and the absence of interference due to a second or a third antibody coat, make this assay useful for analyzing and quantitating monoclonal antibodies obtained by hybridoma or B-cell immortalization.

Biological markers of HIV infection were studied in 17 asymptomatic HIV seropositive subjects in the 12 months preceding the onset of the disease. No single marker of HIV infection preceded the development of AIDS. Therefore, the clinical care of asymptomatic seropositive subjects should include a number of tests to evaluate HIV activity and immune suppression.

Serum from 408 subjects drawn at random from a population of 29,985 was tested by quantitative electrophoresis and, when a narrow band was detected, by immunoelectrophoresis. Six cases of monoclonal gammapathy (MG) were found, including four with IgMK and two with IgGK. The incidence of MG (1.47%) was not particularly high, but the proportion of IgM (66.67%) was abnormally elevated. This peculiarity does not seem due to the advanced age of the subjects tested (58 +/- 13 years) but might be the result of too much inbreeding until recent years.

The distribution in heavy chain classes and light chain types of M components were studied in 757 cases of monoclonal gammapathies. These gammapathies were classified according to clinical and hematological data in 439 myeloma (MM), 165 Waldenström's macroglobulinemia (WM) and 152 monoclonal gammapathies occurring in other conditions. The IgG/IgA ratio differs in myeloma and in non-myelomatous gammapathies (64% IgG versus 33% IgA in myeloma and 95% IgG versus 5% in absence of myeloma). Presence of free light chains in patient's urines (Bence Jones proteinuria) was detected in about 72% of cases of MM, 48% of WM and only in 9% in others gammapathies. Concerning the sex of patients, an equal repartition between males and females is observed in MM, whereas males predominate in WM. The age distribution of the patients shows that 74% of myeloma and 79% of WM were above sixty. In addition to the 757 cases under study, 10 sera with two M Components were characterized.