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PubMed2004

Study of thermal aggregation of oat globulin by laser light scattering.

The heat-induced aggregation of oat globulin was studied using size-exclusion chromatography (SEC) combined with on-line multiangle laser light scattering (MALLS) and quasi-elastic light scattering (QELS). The unheated oat globulin exists as a mixture of hexamer (>95%), trimer, and dimer forms of hexamer. The molecular weight of the hexamer was estimated by MALLS to be 330 000, close to that deduced from the genomic cloned data of the acidic and basic polypeptides of oat globulin. From QELS measurements, it can be predicted that the hexamer exists as two annular trimeric rings, with a diameter of 11.8 nm, placed on top of each other, forming an oblate cylinder with a height of about 8.5 nm. Upon heating at 100 degrees C, the oat globulin hexamers and trimers were dissociated into monomers. The heat-denatured monomers, probably assuming an extended structure, were associated to form small aggregates, which were further aggregated to high molecular weight complexes. Upon further heating (60 min), the soluble aggregates were associated to form insoluble aggregates. Aggregation of oat globulin occurred at a much faster rate at 110 degrees C. The results indicate that the SEC-MALLS-QELS system is suitable for studying thermal aggregation of food proteins.

Avena↗
PubMed2003

Simultaneous determination of human serum albumin, gamma-globulin, and glucose in a phosphate buffer solution by near-infrared spectroscopy with moving window partial least-squares regression.

Near-infrared (NIR) spectra in the 12,000-4,000 cm(-1) region were measured for phosphate buffer solutions containing human serum albumin (HSA), gamma-globulin, and glucose with various concentrations at 37 degrees C. Five levels of full factorial design were used to prepare a sample set consisting of 125 samples of three component mixtures. The concentration ranges of HSA, gamma-globulin and glucose were 0.00-6.00 g dl(-1), 0.00-4.00 g dl(-1) and 0.00-2.00 g dl(-1), respectively. The 125 sample data were split into two sets, the calibration set with 95 data and the prediction set with 30 data. The most informative spectral ranges of 4648-4323, 4647-4255 and 4912-4304 cm(-1) were selected by moving window partial least-squares regression (MWPLSR) for HSA, [gamma]-globulin, and glucose in the mixtures, respectively. For HSA, the correlation coefficient (R) of 0.9998 and the root mean square error of prediction (RMSEP) of 0.0289 g dl(-1) were obtained. For [gamma]-globulin, R of 0.9997 and RMSEP of 0.0252 g dl(-1) were obtained. The corresponding statistic values of glucose were 0.9997 and 0.0156 g dl(-1), respectively. These statistical values obtained by MWPLSR are highly significant and better than those calculated by using the regions reported in the literature. The results presented here show that MWPLSR can select the informative regions with a simple procedure and increase the power of NIR spectroscopy for simultaneous determination of the concentrations of HSA, [gamma]-globulin and glucose in the mixture systems.

Blood Glucose↗
PubMed1965

The disulphide bonds of human and rabbit gamma-globulins.

1. The reaction of the disulphide bonds of the predominant species of human and rabbit gamma-globulins (the 7s gamma-globulins) with sulphite was studied in the presence and absence of denaturing agents and heavy-metal reagents. 2. The total number of bonds reacting/mol. of mol.wt. 160000 was approx. 18 for human and 20 for rabbit gamma-globulin. 3. Six S.S bonds/mol. of human and 6.5 S.S bonds/mol. of rabbit gamma-globulin reacted with sulphite alone at pH6. These appeared to include all the interchain S.S bonds. 4. The number of free SH groups was less than 0.2/mol. of human and less than 0.3/mol. of rabbit gamma-globulin.

Journal Article↗
PubMedDate not supplied

Intravenous immune globulin for patients alloimmunized to random donor platelet transfusion.

Immune globulin, prepared as a chemically and enzymatically unmodified solution in 10 percent maltose at pH 4.25, was administered intravenously, at a dose of 0.4 g per kg per day for five consecutive days, to seven alloimmunized patients who had acute nonlymphocytic leukemia. All patients had an approximately threefold rise in IgG level. Five patients showed no change in lymphocytotoxic antibody (LCTAb) activity and no response to random donor platelets that were administered after the immune globulin. The activity of LCTAb either disappeared or diminished after immune globulin infusion in two patients, and they had borderline acceptable corrected count increments 1 hour after transfusion of pooled random-donor platelet transfusions. Because of the diminished LCTAb activity, the improvements in response to pooled random-donor platelet transfusions cannot be ascribed to the administration of immune globulin. High-dose intravenous immune globulin has not been shown to be effective in reversing the effects of alloimmunization.

Acute Disease↗
PubMed1983

Antithymocyte globulin treatment in patients with aplastic anemia: a prospective randomized trial.

We evaluated the efficacy of antithymocyte globulin for the treatment of moderate to severe aplastic anemia in a randomized controlled study. Eleven of 21 patients initially randomized to receive antithymocyte globulin (given intravenously on eight consecutive days) had sustained improvement in hematopoiesis within three months of treatment; none of 21 control patients who received supportive care alone improved (P = 0.0005). Six of 12 control patients who subsequently received antithymocyte globulin improved. Responders had gradual improvement in hematopoiesis, but none recovered completely normal peripheral-blood counts. The severity of bone-marrow failure, age, cause of aplastic anemia, and transfusion history had no apparent bearing on treatment outcome. The interval from diagnosis to antithymocyte globulin treatment correlated inversely with the chance of a treatment response, although this correlation was not statistically significant. These data indicate that antithymocyte globulin is effective in improving hematopoiesis in some patients with aplastic anemia.

Adolescent↗
PubMed1991

Treatment of aplastic anemia with antilymphocyte globulin and methylprednisolone with or without cyclosporine. The German Aplastic Anemia Study Group.

BACKGROUND AND METHODS: Immunosuppression is the most effective treatment for patients with aplastic anemia, except for bone marrow transplantation. The best results are achieved with antilymphocyte globulin or cyclosporine. Patients have been treated successfully with a combination of both agents, but there has been no controlled evaluation of its efficacy. We conducted a randomized, multicenter trial in 84 patients not eligible for bone marrow transplantation, comparing treatment with antilymphocyte globulin and methylprednisolone (41 patients--the control group) with antilymphocyte globulin, methylprednisolone, and cyclosporine (43 patients--the cyclosporine group). RESULTS: At three months significantly more patients in the cyclosporine group had a complete or partial remission in response to treatment than did patients in the control group (65 percent vs. 39 percent, P less than 0.03); this difference was confirmed at six months (70 percent vs. 46 percent, P less than 0.05). The superior results of the regimen including cyclosporine were most evident in the patients with severe or very severe aplastic anemia, whose response rate at six months was 65 percent, as compared with 31 percent of such patients in the control group (P less than 0.02). Granulocyte and hemoglobin levels became normal in most patients who responded, but platelet counts continued to be subnormal in 61 percent of the patients. Ten of 52 patients with responses (3 in the cyclosporine group and 7 in the control group) relapsed 4 to 37 months after treatment. The actuarial survival of all patients at 41 months is 64 percent in the cyclosporine group and 58 percent in the control group (P = 0.16); among the patients with severe or very severe disease, survival is 80 percent and 44 percent, respectively (P = 0.077). Cyclosporine had substantial but reversible side effects. CONCLUSIONS: Immunosuppressive treatment of aplastic anemia with antilymphocyte globulin, methylprednisolone, and cyclosporine appears to be more effective than a regimen of antilymphocyte globulin and methylprednisolone without cyclosporine and may thus represent a treatment of choice for patients who are not eligible for bone marrow transplantation.

Adult↗
PubMed1994

Hepatitis C infection in patients with primary hypogammaglobulinemia after treatment with contaminated immune globulin.

BACKGROUND: In Scandinavia many patients with primary hypogammaglobulinemia contracted non-A, non-B hepatitis after intravenous treatment with an immune globulin product that was later found to contain a non-A, non-B hepatitis virus. METHODS: We studied the prevalence and clinical course of hepatitis C virus (HCV) infection in a group of 55 Norwegian patients with primary hypogammaglobulinemia and investigated its association with the use of contaminated immune globulin. We used the polymerase chain reaction to detect HCV RNA and performed HCV genotyping. We also analyzed the responses to treatment with interferon. RESULTS: Of 20 patients who received the contaminated immune globulin, 17 were seropositive for HCV RNA: In addition, 1 of 35 patients not exposed to the contaminated immune globulin was HCV RNA--positive. HCV genotype V was found in all 12 patients for whom genotyping was performed, but 8 patients also had genotype II or III, or both. All HCV RNA--positive patients had abnormal results on biochemical liver tests. All liver-biopsy specimens (from 15 patients) were abnormal, with portal inflammation, bile-duct damage, and focal necrosis. In six patients there was cirrhosis. Two patients died of liver failure. In 4 of the 10 patients treated with interferon there were complete, though transient, biochemical responses, but the follow-up biopsy specimens showed evidence of histologic progression. The poorest responses to interferon were among the patients with multiple HCV genotypes. All but one patient remained positive for HCV RNA: CONCLUSIONS: In patients with primary hypogammaglobulinemia there was a high rate of HCV infection after treatment with contaminated immune globulin. In these immunocompromised patients HCV infection has a severe and rapidly progressive course, and responses to interferon are poor.

Adolescent↗
PubMed2006

Treatment of pemphigus vulgaris with rituximab and intravenous immune globulin.

BACKGROUND: Pemphigus vulgaris is a potentially fatal autoimmune mucocutaneous blistering disease. Conventional therapy consists of high-dose corticosteroids, immunosuppressive agents, and intravenous immune globulin. METHODS: We studied patients with refractory pemphigus vulgaris involving 30% or more of their body-surface area, three or more mucosal sites, or both who had inadequate responses to conventional therapy and intravenous immune globulin. We treated the patients with two cycles of rituximab (375 mg per square meter of body-surface area) once weekly for 3 weeks and intravenous immune globulin (2 g per kilogram of body weight) in the fourth week. This induction therapy was followed by a monthly infusion of rituximab and intravenous immune globulin for 4 consecutive months. Titers of serum antibodies against keratinocytes and numbers of peripheral-blood B cells were monitored. RESULTS: Of 11 patients, 9 had rapid resolution of lesions and a clinical remission lasting 22 to 37 months (mean, 31.1). All immunosuppressive therapy, including prednisone, could be discontinued before ending rituximab treatment in all patients. Two patients were treated with rituximab only during recurrences and had sustained remissions. Titers of IgG4 antikeratinocyte antibodies correlated with disease activity. Peripheral-blood B cells became undetectable shortly after initiating rituximab therapy but subsequently returned to normal values. Side effects that have been associated with rituximab were not observed, nor were infections. CONCLUSIONS: The combination of rituximab and intravenous immune globulin is effective in patients with refractory pemphigus vulgaris.

Adolescent↗
PubMed2004

Neutralizing antibodies to hepatitis C virus (HCV) in immune globulins derived from anti-HCV-positive plasma.

The role of humoral immunity in hepatitis C virus (HCV) infections is uncertain. Nevertheless, there is increasing evidence for neutralizing antibodies to HCV in the serum or plasma of chronically infected individuals. Immune globulins prepared by ethanol fractionation of plasma had long been considered safe until a commercial immune globulin product, Gammagard, prepared from plasma from which units containing anti-HCV had been excluded, transmitted HCV to recipients. Studies suggested that the exclusion might have removed neutralizing antibodies from the plasma and hence compromised the safety of the resulting immune globulins. In the present study, by using chimpanzees and a recently validated in vitro system based on neutralization of infectious HCV pseudoparticles, we found broadly reactive neutralizing and protective antibodies in experimental immune globulin preparations made from anti-HCV-positive donations. Neutralizing antibodies were also found in Gammagard lots made from unscreened plasma that did not transmit hepatitis C but not in Gammagard lots, which were prepared from anti-HCV-screened plasma, that did transmit hepatitis C. The results provide an explanation for the mechanism by which the safety of this product was compromised. Immune globulins made from anti-HCV-positive plasma and containing broadly reactive neutralizing antibodies may provide a method of preventing HCV infection.

Animals↗
PubMed1961

Studies on fluorescent antibody staining. I. Non-specific fluorescence with fluorescein-coupled sheep anti-rabbit globulins.

1. A study has been made of the non-specific fluorescent staining of splenic imprints treated with fluorescent sheep antibody globulins. 2. In tissue imprints made with the spleens of antigen-stimulated animals, no morphological distinction was evident between areas showing non-specific fluorescence and specific fluorescence. 3. Elimination of non-specific fluorescence was not achieved by any one, or any combination of the following: (a) conjugating only gamma globulins with fluorescein isothiocyanate; (b) removal of dialyzable fluorescent products on sephadex, followed by concentration through the use of pressure dialysis; (c) use of crystalline preparations of fluorescein isothiocyanate. 4. Individual preparations of fluorescent antibodies were separated by gradient elution chromatography on diethylaminoethyl (DEAE) cellulose into fractions possessing different numbers of fluorescein radicals per molecule of globulin. 5. The coupling ratio of 50 mg fluorescein isothiocyanate (FITC) per gm of protein, as commonly advocated, can not be recommended for the precise localization of antibody globulin in tissues owing to the capacity of the coupled products to give non-specific fluorescent staining. When crystalline preparations of FITC are used instead of the amorphous product at 50 mg/gm protein, far too high non-specific fluorescence results. 6. A fraction with bright specific fluorescence and no or negligible nonspecific fluorescence was obtained from each fluorescent antibody that was prepared by using 6 to 8 mg of crystalline fluorescein isothiocyanate per gm of globulin and was then subjected to DEAE-cellulose chromatography and gradient elution to eliminate the most highly coupled molecules.

Animals↗
PubMed1968

Cell to cell interaction in the immune response. IV. Site of action of antilymphocyte globulin.

In this series of papers it has been shown that the immune response of mice to sheep erythrocytes requires the participation of two classes of lymphoid cells. Thymus-derived cells initially react with antigen and then interact with another class of cells, the antibody-forming cell precursors, to cause their differentiation to antibody-forming cells. Antilymphocyte globulin depressed the ability of mice to respond to sheep erythrocytes. This effect was more marked when the antigen was injected intraperitoneally than intravenously, and occurred only when the antilymphocyte globulin was given before or simultaneously with antigen. Injection of thymus cells restored to near normal the ability to respond to an intravenous injection of sheep erythrocytes. Spleen cells from antilymphocyte globulin-treated mice gave a weak adoptive immune response in irradiated recipients. The addition of thymus cells however enabled a response similar to that given by normal spleen cells. When thymectomized irradiated recipients were used, normal spleen cells continued to give a higher response to a challenge of sheep erythrocytes at 2 and 4 wk postirradiation than did spleen cells from ALG-treated donors. This result is more consistent with the notion that thymus-derived target cells are eliminated, rather than temporarily inactivated, by antilymphocyte globulin. These findings suggest that, in vivo, antilymphocyte globulin acts selectively on the thymus-derived antigen-reactive cells.

Animals↗
PubMed1952

Experimental globulin glomerulonephritis in rabbits; morphological and functional changes.

Diffuse glomerulonephritis was induced in unilaterally nephrectomized albino rabbits by giving them two massive intravenous injections of purified bovine serum gamma globulin. Morphologically, these renal lesions were similar to those of human acute and subacute glomenilonephritis. Globulin nephritis in rabbits also closely resembles experimental nephritis induced by injection of other foreign proteins. The course of this experimental disease was characterized by abrupt onset of proteinuria, hematuria, elevation of serum globulin, and reduction of albumin. Uremia developed in some animals. The manifestations of nephritis during life coincided with the development of demonstrable antibodies to the injected globulin. In view of the similarities between experimental globulin nephritis in rabbits and human nephritis, together with common features with other experimental nephritis, it is suggested that all may have a common pathogenetic mechanism. It is concluded that quantitative antibody response may be of greater importance in the pathogenesis of nephritis than antigen specificity.

Animals↗
PubMed1990

A community-wide outbreak of hepatitis A in a religious community: impact of mass administration of immune globulin.

Community-wide outbreaks of hepatitis A are frequently prolonged and difficult to control. An extensive outbreak of hepatitis A in a religious community provided an opportunity to assess the effect of mass administration of immune globulin on the course of the outbreak. Between July 1, 1988 and May 30, 1989, 204 cases occurred among 3,500 residents (58/1,000), with persons aged 5-19 years having the highest attack rate. It was found that 89% of persons older than age 19, but no persons under age 20, had evidence of prior hepatitis A infection. During a 5-day campaign, immune globulin (0.02 ml/kg) was administered to 2,287 (65%) of the 3,500 residents. The cost of vaccine and syringes was less than $3,500. New cases among immune globulin recipients virtually stopped 2 weeks after the campaign, and the incidence of hepatitis in the community decreased from 9.6/week to 1.9/week. Among persons younger than age 20 years, the efficacy of immune globulin was 88.9% (95% confidence interval 77.9-94.5) for seven months. Although the authors cannot be sure that the outbreak will not recur, they believe that mass administration of immune globulin appears to have been partially effective at controlling this community-wide outbreak.

Adolescent↗
PubMed1974

Globulin-specific Proteolytic Activity in Germinating Pumpkin Seeds as Detected by a Fluorescence Assay Method.

The proteolytic activities of alpha-chymotrypsin, trypsin, pepsin, bromelain, and an extract from germinating pumpkin seeds (Cucurbita moschata) were determined by their ability to effect the release of 1-anilino-8-naphthalenesulfonate bound to internal hydrophobic sites in intact protein substrates. Casein, glyceraldehyde-3-P dehydrogenase, urease, catalase, pumpkin seed globulin, and bovine serum albumin enhanced the fluorescence of 1-anilino-8-naphthalenesulfonate sufficiently to be used as proteolytic substrates. Chymotrypsin, trypsin, pepsin, and bromelain exhibited activity against all or almost all of the protein substrates. The activity of 1 mug of alpha-chymotrypsin or trypsin and 100 ng of pepsin could be easily detected by this method of assay within 4 to 5 minutes depending upon the substrate. The enzyme extracted from 3-day germinated pumpkin seeds exhibited strong activity only against pumpkin seed globulin, weak activity against the globulins of squash and cucumber and casein, and no activity against the other protein substrates. Activity against pumpkin globulin was maximal at pH 7.4. When assayed by an increase in ninhydrin-positive products, the enzyme extract from pumpkin seeds also showed strong activity against pumpkin globulin and weak activity against casein. The 1-anilino-8-naphthalenesulfonate-fluorescence method was at least 20 times more sensitive than the ninhydrin method and was 10 to 20 times more rapid.

Journal Article↗
PubMed1983

Oat seed globulin: subunit characterization and demonstration of its synthesis as a precursor.

The predominant storage protein of oat (Avena sativa L.) seeds is a saline-soluble globulin with a mol wt of 320,000 which is composed of six large (M(r) = 35,000 to 40,000) and six small (M(r) = 20,000 to 25,000) subunits. Experiments were conducted to further describe the subunit polypeptides and to identify the initial translation products of globulin mRNAs. Approximately 20 large subunits and 10 small subunits were resolved by two-dimensional gel analysis. The large and small subunits had acidic and basic isoelectric points, respectively. Disulfide-linked complexes of one large and one small subunit were isolated by extraction in buffer lacking a reducing agent. The NH(2)-terminal sequence of the small subunits was homologous to a small subunit of soybean glycinin. Immunoprecipitation of in vitro translation products of poly(A)(+) RNA with anti-oat globulin sera yielded M(r) = 60,000 to 68,000 polypeptides. In vivo labeling of spikelets with radioactive amino acids resulted in high amounts of incorporation into polypeptides with M(r) = 65,000 to 68,000 which were immunoprecipitated with anti-globulin sera. These two results suggest oat globulin is synthesized as a higher mol wt precursor which is subsequently processed to yield the large and small subunit polypeptides.

Journal Article↗
PubMed1984

Quantitation of oat globulin by radioimmunoassay.

A radioimmunoassay was used to determine the globulin content of developing oat (Avena sativa L. var Coker 6622) seeds. The globulin content increased from 0.32 to 2.37 milligrams per seed from 2 to 21 days post anthesis. The amount of globulin did not increase from 21 to 30 days post anthesis. When the total protein of seeds harvested 21, 24, and 30 days post anthesis was measured by micro-Kjeldahl analysis, it was determined to be 3.15 milligrams per seed for each sampling date. When the amount of globulin per seed was compared to this value, the relative proportion of globulin to total seed protein was approximately 75%.

Journal Article↗
PubMed1987

Biochemistry of Fern Spore Germination: Globulin Storage Proteins in Matteuccia struthiopteris L.

Two globulin storage proteins have been identified in spores of the ostrich fern, Matteuccia struthiopteris (L.) Todaro. The two proteins comprise a significant amount of the total spore protein, are predominantly salt-soluble, and can be extracted by other solvents to a limited extent. The large 11.3 Svedberg unit (S) globulin is composed of five polypeptides with molecular weights of 21,000, 22,000, 24,000, 28,000 and 30,000. Each polypeptide has several isoelectric point (pI) variants between pH 5 and 7. The small 2.2S storage protein has a pI > 10.5 and is composed of at least two major polypeptides of 6,000 and 14,000 M(r). The amino acid composition of both storage proteins reveals that the 11.3S protein is particularly rich in aspartic and glutamic acid, while the 2.2S protein has few acidic amino acids. During imbibition and germination the globulin fraction declines rapidly, with a corresponding degradation of individual polypeptides of each protein. Polyclonal antibodies against each of the two proteins were produced and used for immunolocalization to determine the site of storage protein deposition within the quiescent spore. The proteins were sequestered in protein bodies of 2 to 10 micrometers, that are morphologically similar to those found in the seeds of flowering plants. The results suggest that spore globulins are biochemically similar to seed globulins, especially those found in some cruciferous seeds.

Journal Article↗
PubMed1983

Antithymocyte globulin treatment of severe aplastic anaemia.

Nineteen patients with severe aplastic anaemia were treated with antithymocyte globulin. Ten patients obtained remissions (transfusion independent, at least 45000 platelets and 2000 PMN/mm3) within 2-3 months and continue in remission 5-35 months after antithymocyte globulin. Ages of responders ranged from 17 to 71. Complications of antithymocyte globulin included arthralgias, rash, serum sickness, angioedema and fever. Two patients died during, two shortly after, and one 10 months after therapy. One patient with a previous remission following antithymocyte globulin relapsed and achieved a second remission with retreatment. Previous androgen therapy did not affect outcome since two of four patients with and eight of 15 patients without previous androgen therapy achieved remission with ATG. Treatment with antithymocyte globulin is a promising alternative to bone marrow transplantation in the treatment of severe aplastic anaemia.

Adolescent↗