Anti-human Hemoglobin Research Articles

How Long to Abstain from Eating Red Meat before Fecal Occult Blood Tests

Brief Reports1 September 1990How Long to Abstain from Eating Red Meat before Fecal Occult Blood TestsElliot J. Feinberg, MD, William M. Steinberg, MD, Bonnie L. Banks, BA, James P. Henry, BSElliot J. Feinberg, MDSearch for more papers by this author, William M. Steinberg, MDSearch for more papers by this author, Bonnie L. Banks, BASearch for more papers by this author, James P. Henry, BSSearch for more papers by this authorAuthor, Article, and Disclosure Informationhttps://doi.org/10.7326/0003-4819-113-5-403 SectionsAboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinkedInRedditEmail ExcerptHemoccult's long recognized use for colon cancer screening (1, 2) has recently been challenged by newer fecal occult blood tests with reputed advantages in sensitivity and specificity.The HemoQuant quantitatively measures the concentration of heme-derived porphyrins (3, 4). The Hemoccult SENSA, a modified guaiac-based test, reportedly detects lower levels of hemoglobin, thereby increasing sensitivity (5). The HemeSelect, using an antihuman hemoglobin antibody, is reported to be highly sensitive and specific (6, 7). Although dietary constituents, especially red meat, can increase the false-positive rate of the Hemoccult (1, 2, 8, 9), Hemoccult SENSA (5), and HemoQuant (4) tests, the comparative duration...

Liver and Kidney Injury After Administration of Hemoglobin Cross-Linked with Bis(3,5-Dibromosalicyl) Fumarate

Human hemoglobin cross-linked between the alpha chains with bis (3,5-dibromosalicyl) fumarate (DBBF-Hb) was exchange transfused in swine and the histomorphologic changes were evaluated. Following exchange, animals were euthanized and tissues were taken for light and electron microscopy at 7.5 hours and days 1, 4, 7, and 15. Consistent hepatocellular and renal epithelial cell changes were seen. Hepatic injury, evident at 7.5 hours as cellular vacuolization, progressed to necrosis and acute inflammatory cell infiltration by days 1 and 4, was resolving by 7 days and was completely resolved by day 15. Cytochemical stains for iron and hemoglobin revealed positive material in Kupffer cells, endothelial cells, and necrotic hepatocytes. Rabbit anti-human hemoglobin antibody staining revealed immunoreactive material diffusely present at days 1 and 4 and limited to solitary hepatocytes by day 15. Kidney injury began as proximal tubular epithelial vacuolization and intraluminal casts progressing to tubular necrosis by 24 hours, with resolution by day 15. Iron and hemoglobin stains demonstrated these materials in the early lesions. Immunocytochemistries demonstrated human hemoglobin that remained as late as day 15. Electron microscopy revealed degeneration and regeneration of epithelial cells. The renal lesions were consistent with hemoglobinuria. The liver lesion was less well defined but was self limited.

第15回日本総合健診医学会学術大会講演抄録 抗ヒトーヘモグロビン抗体を用いた便潜血反応の基礎的検討およびドック受診者における成績について

第15回日本総合健診医学会学術大会講演抄録 抗ヒトーヘモグロビン抗体を用いた便潜血反応の基礎的検討およびドック受診者における成績について

Discrepancy in the expression of autoantibodies in healthy aged individuals

Sera from 50 healthy old subjects and from 51 young controls were tested by ELISA assays for a panel of autoantibodies, including IgM RF, anti-DNA, anti-F(ab′) 2, anti-thyroglobulin, anti-human albumin, anti-human hemoglobin, anti-secretory component from human IgA, and anti-gliadin. In vitro production of anti-DNA as well as anti-F(ab′) 2 antibodies were measured after stimulation of PBMC by pokeweed mitogen (PWM) in 12 healthy elderly subjects and 11 young controls. Sera from elderly donors contained threefold higher amounts of IgM RF than young controls ( P < 0.001). On the contrary, the levels of anti-DNA as well as anti-F(ab′) 2 antibodies were similar in both groups ( P < 0.3 for the two determinations). Anti-DNA and anti-F(ab′) 2 antibodies were also measured in supernates of PWM-stimulated glass nonadherent PBMC cultures from both old and young healthy donors without finding any significant difference between the two groups. Additional ELISA tests were also performed in both elderly and young control sera to detect antibodies against six other different antigens. No significant difference was found in the percentages of positive sera between the two groups. This discrepancy in production of IgM RF compared to other autoantibodies in healthy elderly subjects does not provide support for a general increase of autoantibodies with aging. The biological significance of an increase in IgM RF with aging remains to be determined.

抗ヒトヘモグロビン抗体を用いた免疫学的便潜血反応の検討

消化管出血の検索のため従来から行われている化学的便潜血反応には,ヒトヘモグロビン以外の物質と非特異的反応をおこすという欠点がある.この問題を解決するため抗ヒトヘモグロビン抗体を作製し,二次元両者拡散法(Ouchterlony法)を用いて免疫学的便潜血反応を臨床的に検討した.非消化管疾患107例では9.3%,消化管疾患150例をは48.7%,大腸癌60例では75.0%に陽性を示し,健常者30例では全例で陰性であった.Ouchterlony法は消化管疾患や大腸癌では,化学的便潜血反応であるシオノギスライドB法とほぼ同様の陽性率を示したが,非消化管疾患および健常者では明らかに低率を示し,偽陽性率の改善が可能であった.従って本法は消化管疾患による出血,特に大腸癌からの出血のスクリーニング法として有用と考えられた.

Synthesis of adult-type hemoglobin in human erythremia cell line

KMOE -2/05 cells, derived from a patient with acute erythremia, became benzidine-positive after the addition of cytosine arabinoside (CA). Radioimmunoassays using antihuman hemoglobin antibodies revealed an elevated amount of hemoglobin in the CA-exposed cells over that in the control cells (without CA). Isoelectric focusing of the CA-exposed cell lysate formed benzidine-positive foci in the positions of human adult Hb (HbA) and human fetal Hb (HbF). To determine the types of globin synthesized in the CA-exposed cells, globin chains internally labeled with [3H] leucine were purified by carboxy-methyl (CM)-Sephadex column chromatography, immunoadsorption by Sepharose-coupled antihuman Hb antibodies and Sephadex G-100. The labeled globin chains were finally separated by CM-cellulose chromatography in urea. Two distinct peaks of radioactivity were shown in exactly the same fractions as carrier human globin alpha- and beta-chains. These observations indicate that these KMOE -2/05 cells synthesize HbA.

Synthesis of Adult-Type Hemoglobin in Human Erythremia Cell Line

KMOE -2/05 cells, derived from a patient with acute erythremia, became benzidine-positive after the addition of cytosine arabinoside (CA). Radioimmunoassays using antihuman hemoglobin antibodies revealed an elevated amount of hemoglobin in the CA-exposed cells over that in the control cells (without CA). Isoelectric focusing of the CA-exposed cell lysate formed benzidine-positive foci in the positions of human adult Hb (HbA) and human fetal Hb (HbF). To determine the types of globin synthesized in the CA-exposed cells, globin chains internally labeled with [3H] leucine were purified by carboxy-methyl (CM)-Sephadex column chromatography, immunoadsorption by Sepharose-coupled antihuman Hb antibodies and Sephadex G-100. The labeled globin chains were finally separated by CM-cellulose chromatography in urea. Two distinct peaks of radioactivity were shown in exactly the same fractions as carrier human globin alpha- and beta-chains. These observations indicate that these KMOE -2/05 cells synthesize HbA.

The Identification of Human Bloodstains by Means of Thin-Layer Immunoassay: A Preliminary Report

The application of thin-layer immunoassay to the problem of human bloodstain identification is described. The differentiation of human, dog and rabbit bloodstains has been achieved by this method with the use of anti-human haemoglobin serum.

The Identification of Human Bloodstains by Means of a Micro-Thin-Layer Immunoassay Procedure

A method for the identification of human bloodstains using a micro-thin-layer immunoassay (TIA) procedure is presented. The equivalent of approximately 0.01 micro /leter of human blood may be detected using this method with anti-human hemoglobin. Blind studies and stability studies indicate that the method is sufficiently specific and sensitive to be of forensic science value.

Hindrance in the simultaneous binding of two distinctly separate antibody populations to the hemoglobin antigen

Two subpopulations of goat anti-human hemoglobin a 1 (HbA 1) Fab' monomers were isolated. One binds at the α subunit of hemoglobin and not at the β subunit. The other binds only at the β subunit of hemoglobin. The total number of antigenic sites on hemoglobin bound by the two Fab' subpopulations exceeds the number of antigenic sites bound by an equimolar mixture of the two populations. Also, the amounts of each Fab' subpopulation needed to reach equivalence point when titrating HbA 1 already bound to the other Fab' were 25% less than the amounts of Fab' needed to reach equivalence point when titrating HbA 1 alone. These results indicate that part of the Fab' present in each of the two subpopulations interfere with one another in binding antigen. These particular Fab' monomers bind at the α and β subunits of HbA 1 either at adjacent but independent determinant sites or at nonadjacent sites, the conformations of which are affected by the binding of Fab' at the other antigenic site.

The fate of cytoplasmic vanadium. Implications on (NA,K)-ATPase inhibition.

The fate of vanadate (+5 oxidation state of vanadium) taken up by the red cell was studied using EPR spectroscopy. The appearance of an EPR signal indicated that most of the cytoplasmic vanadate is reduced to the +4 oxidation state with axial symmetry characteristic of vanadyl ions. The signal at 23 degrees C was characteristic of an immobilized system indicating that the vanadyl ions in the cytoplasm are associated with a large molecule. [48V]Vanadium eluted with hemoglobin when the lysate from Na3[48V[O4-treated red cells was passed through a Sephadex G-100 column and rabbit anti-human hemoglobin serum caused a hemoglobin-specific precipitation of 48V when added to the red cell lysate. Both results indicate that hemoglobin is the protein which binds cytoplasmic vanadyl ions. However, neither sodium vanadate nor vanadyl sulfate bound to purified hemoglobin in vitro. Finally, transient kinetics of vanadyl sulfate interaction with the sodium-and potassium-stimulated adenosine triphosphatase showed that the +4 oxidation state of vanadium is less effective than the +5 oxidation state in inhibiting this enzyme. These results indicate that oxidation-reduction reactions in the cytoplasm are capable of relieving vanadate inhibition of cation transport.

Properties of goat anti-human hemoglobin antibodies fractionated on subunit affinity columns

The antibodies to human hemoglobin A, (HbA 1) in the sera of two goats were fractionated on the basis of their ability or inability to bind to the isolated α and β subunits of the hemoglobin. The fractionation was carried out by a series of affinity chromatography steps and the characterization of the binding of the antibody populations thus obtained to HbA 1, α and β revealed some predominating characteristics common to the two goat sera. In roughly proportional yields, the anti-HbA 1 in the sera were fractionated into three major catergories: antibodies which bind only to HbA 1 and β chains, antibodies which bind only to HbA 1 and α chains and antibodies which bind to HbA 1 and both α and β chains. One goat also had an antibody population that would bind HbA 1 but neither α nor β. There was, in both sera, a larger proportion of α binding control to β binding antibodies and of α−ß non-cross-reacting compared to α−ß cross-reacting antibodies. Upon consideration of the amino acid sequence and three-dimensional structures of both the hemoglobin antigen and the host's own hemoglobin with respect to sequence differences and the exposure and clustering of such differences, the existence and proportions of the diffrent antibody populations can be explained.

The inhibition of the primary interaction between 1 2 5I-labeled human hemoglobin and rabbit anti-human hemoglobin sera: A sensitive radioimmunoassay technique

A rapid and sensitive radioimmunoassay technique for hemoglobin based upon a modification of the Farr test is described. Since hemoglobin is soluble in ammonium sulfate whereas antibody is precipitated, the amount of free and antibody-bound radiolabeled human hemoglobin can be quantitatively measured after ammonium sulfate precpitation. Nanogram quantities of hemoglobin can be detected by the addition of unlabeled human hemoglobin to the antiserum before the labeled hemoglobin, thus establshing a competition reaction between labeled and unlabeled antigen for antibody. If sufficient quantities of unlabeled human hemoglobin are present, all of the antibody combining sites become saturated and the primary interaction between 1 2 5I-labeled hemoglobin and antibody is completely inhibited. The method can be employed to discriminate various normal human hemoglobins, non-human primate hemoglobins as well as sickle-cell hemoglobin (HbS). Other possible applications of the technique are discussed.

The use of anti-human haemoglobin in forensic serology.

An assessment of a commercial anti-human haemoglobin reagent has been conducted under experimental conditions and in forensic casework examinations. The immunoglobulin preparation used was shown to react with human haemoglobin and baboon haemoglobin in similar fashion. The sensitivity of the reagent is adequate to establish the presence of human haemoglobin in human bloodstains. The use of anti-human haemoglobin reagents in medicolegal work is advocated.

The immunological identification of foetal haemoglobin in bloodstains in infanticide and associated crimes.

An anti-human foetal haemoglobin reagent has been assessed for the identification of foetal haemoglobin in bloodstains encountered in forensic serology. The results obtained experimentally on known material have been augmented in a ‘blind’ survey involving stains made from 38 separate cord blood samples. These results establish that this reagent when used in conjunction with anti-human haemoglobin reagent can successfully differentiate cord blood stains from those of adult blood. The methods described represent an improvement on those previously available for the identification of foetal haemoglobin in bloodstains.

Pigmenturia from conga drumming. Hemoglobinuria and myoglobinuria.

A young man developed pigmenturia after conga drumming. The pigmented urine contained myoglobin, identified by Ouchterlony immunodiffusion with rabbit anti-human myoglobin immune serum, and hemoglobin, identified by Ouchterlony immunodiffusion with goat anti-human hemoglobin immune serum and by polyacrylamide gel electrophoresis of the hemoglobin-haptoglobin complex. Concentrations of myoglobin and hemoglobin in the patient's urine were approximately 17 µg/ml and 2 mg/ml, respectively. The urine findings, a minimal rise in the serum creatine phosphokinase and glutamic-oxalacetic transaminase levels, a marked fall in serum haptoglobin, and a marked rise in serum hemoglobin during a planned drumming session suggest that myoglobinuria and hemoglobinuria may be caused by the exercise and severe hand trauma associated with conga drumming.

Immunochemical and physicochemical properties of haptoglobin, abtihemoglobin antibody, and their complexes with hemoglobin

The immunochemical properties of haptoglobin which binds specifically with hemoglobin were compared with those of antihemoglobin A 1 antibody. Hemoglobin-haptoglobin shows neither precipitation reaction nor complement fixation reaction under conditions in which hemoglobin-antihemoglobin antibody shows these immunological reactions. From these observations, the absence of lattive formation between hemoglobin and haptoglobin was suggested. Antihuman hemoglobin antibody binds deoxygenated human hemoglobin and hemoglobin-haptoglobin complex, both of which human haptoglobin does not. On the other hand, although human haptoglobin binds hemoglobins of horse, rabbit, and frog, antihuman hemoglobin antibody does not. And from the data on circular dichroism spectra of hemoglobin-haptoglobin complex and hemoglobin-antihemoglobin antibody complex, the differences in environmental structure around the heme between hemoglobin-haptoglobin complex and hemoglobin-antihemoglobin antibody complex are clarified which may have some connection with the differences in immunochemical reactions between these two complexes.