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Seasonal variations of blood haptoglobin level of brown bears in Japan.

Haptoglobin (Hp), a hemoglobin-binding protein, is known as an acute phase protein increasing in blood during inflammation in most mammals. On the basis of our previous studies on purification and characterization of bear Hp (Comp. Biochem. Physiol. 110B, 785-789, 1995), in this study, we developed an immunoassay method to measure serum Hp level in bear, and measured the concentration of Hp in blood samples collected from 84 reared and 25 wild brown bears in Hokkaido, Japan. The mean serum Hp concentration was 0.94 +/- 0.25 mg/ml in wild bears, which is nearly equal to those reported in other species. In reared bears, the Hp concentration was apparently higher (3.82 +/- 0.29 mg/ml), although total protein and albumin concentrations were nearly equal in the two groups. A significant seasonal variation of serum Hp, low in spring and high in autumn and winter, was found in reared bears. Possible factors participating in the seasonal variation were discussed with special references to hibernation.

Animals↗

Parallel changes in the blood levels of abnormally-fucosylated haptoglobin and alpha 1,3 fucosyltransferase in relationship to tumour burden: more evidence for a disturbance of fucose metabolism in cancer.

Levels of an abnormally-fucosylated form of the serum glycoprotein, haptoglobin (FHp) and an enzyme, alpha 1,3 fucosyltransferase (FT) have been measured in blood specimens from women with carcinoma of the ovary or breast who are undergoing chemotherapy. The levels of FHp and FT increased if the women had progressive disease and decreased if they showed complete response to therapy. The statistical correlation between the blood concentrations of these two substances is very strong (P less than 0.0001, chi 2 test). These results and recent studies of fucosyltransferases and cell adhesion molecules from other laboratories, suggest that there are important changes in fucose metabolism in cancer which are worthy of further investigation.

Biomarkers, Tumor↗

Increased fucosylation and other carbohydrate changes in haptoglobin in ovarian cancer.

Cancer sera have high levels of an abnormal form of haptoglobin (Hp) that can be extracted from blood using the fucose-specific lectin, Lotus tetragonolobus. In order to investigate the carbohydrate abnormality that is responsible for this effect, the monosaccharide composition of Hp has been measured in Hp preparations isolated from blood samples of healthy women and women with ovarian cancer. The fucose content (g/100 g protein) of Hp was considerably elevated (7-fold) in ovarian cancer; whereas the concentrations of the other sugars were not significantly increased. There were significant increases in the galactose and N-acetylglucosamine contents of the 'cancer' Hp if the levels were expressed relative to the mannose content. This latter finding suggests that there are more oligosaccharide branches on the 'cancer' Hp. The results indicate that Hp is extracted by lotus lectin from ovarian cancer sera because of the increased fucose levels and provide further support for a disturbance in fucose metabolism in cancer.

Aged↗

Depleted serum haptoglobin in acute bovine trypanosomiasis.

Haemoglobin-binding protein (Haptoglobin, Hp) was assayed in the sera of control and zebu calves with acute T. vivax infection. Hp in control Zebu calves ranged between 123.5 and 236.5 mg dl-1 while Hp in the infected calves was too low to measure or absent. It was suggested that absence of Hp in the infected calves may be as a result of gradual intravascular haemolysis and removal of the Hp-Hb complex by the reticuloendothelial system.

Acute Disease↗

Structure and evolution of artiodactyla haptoglobins.

1. Artiodactyla haptoglobins (Hps), goat, sheep and cattle (family Bovidae), and pig (family Suidae) were structurally characterized. 2. The polymeric Hp systems of goat, sheep and cattle were similar to the polymeric human Hp system, while the monomeric system of pig was more comparable to the monomeric human form. 3. All members of the Artiodactyla (family Bovidae) examined exhibited a large polypeptide subunit, comparable to that of the beta subunit of human Hp. 4. In addition, a small subunit, similar in molecular weight to the human alpha 2 subunit, was demonstrated. Pig Hp was shown to have two subunits, one slightly larger than the human beta subunit and the other intermediate in size to the human alpha 1 and alpha 2 subunits. 5. Immunoelectrophoretic and immunodiffusion studies indicated complete cross reactivity among the polymeric Artiodactyla Hps. 6. The polymeric Hps do not, however, cross react with the monomeric pig Hp.

Animals↗

Catabolism of avian and mammalian haptoglobins and their complexes with haemoglobin in chicken.

The following 125I-labelled preparations of human and chicken haptoglobins (Hph, Hpc) and their complexes with human and chicken haemoglobins (Hph-Hbh, Hpc-Hbc, Hph-Hbc) were injected intravenously to chickens. Hpc exhibited long half-life time (t1/2 = 12.9 hr), whereas the homologous Hpc-Hbc complex disappeared from circulation faster (t1/2 = 3.1 hr). This suggested that the Hpc-Hbc complex was internalized by the receptor-mediated process in the avian liver. Mammalian Hph-Hbh complex was eliminated from chicken circulation rather fast (t1/2 = 0.5 hr) in contrast to "mixed" Hph-Hbc complex which was characterized by the relatively long half-life time (t1/2 = 4.4 hr). Hph-Hbc complex displayed 1-2 sites for binding an anti-Hph antibody i.e. less than Hph-Hbh or Hph-Hbc complexes. The Hph binding with Hbc seems to be weaker than with mammalian haemoglobins.

Animals↗

Plasma haptoglobin in cattle (Bos taurus) exists as polymers in association with albumin.

1. Plasma haptoglobin (Hp) in cattle (Bos taurus) has a molecular mass so large that it is virtually unable to penetrate 4% polyacrylamide gels and is excluded from gel filtration media with an upper exclusion limit of greater than 1000 kDa. 2. In most species, apart from ruminants, Hp has a molecular mass of 100 kDa, consisting of two subunits of 40 kDa and two subunits of 9 kDa, although in a few species, such as man, a genetic variant of Hp forms polymers of higher mass. 3. Bovine Hp was purified by (i) salt precipitation, gel filtration and ion exchange chromatography and (ii) by affinity chromatography and gel filtration, with final purification by (iii) preparative electrophoresis. 4. Bovine Hp could only penetrate sodium dodecyl sulphate polyacrylamide gels following denaturation with mercaptoethanol, when it was shown to be composed of subunits of 40 and 16 kDa, but a protein of 67 kDa was consistently present. 5. Western blotting identified the 67 kDa protein as bovine serum albumin. 6. The polymeric form of Hp found in bovine plasma is formed by association of 40 and 16 kDa subunits with albumin.

Animals↗

Serum haptoglobin appearance during neonatal period is associated with acid phosphatase (ACP1) phenotype.

Erythrocyte acid phosphatase (ACP1) is a polymorphic enzyme found in many tissues and acts in vivo as a flavin-mononucleotide phosphatase. We have recently observed a relation between this enzyme and length of gestation. The present study shows that the pattern of appearance of serum haptoglobin during the neonatal period is associated with ACP1 phenotype suggesting some important function of this polymorphic enzyme in human development.

Acid Phosphatase↗

Ninety-five percent probability of paternity with HLA, ABO and haptoglobins.

Using HLA tissue typing as the basic genetic test along with ABO and haptoglobin typing, it is generally possible either to exclude a putative father conclusively or, in nonexclusion cases, to assign a greater than 95% probability of paternity. In 22 of 53 nonexclusion cases, the putative father had a probability of paternity above 99% based on these genetic tests.

ABO Blood-Group System↗

Haptoglobin typing of human bloodstains using a specific DNA probe.

The stability of DNA in human bloodstains and various post mortem tissues has been investigated. High molecular weight (HMW) DNA was usually recovered from dried bloodstains, even those up to a few years old, but very rapid degradation was found to occur post mortem in the liver, pancreas, spleen and kidney. Other tissues such as the heart, thyroid and skeletal muscle were found to give a reasonable yield of HMW DNA during the first few days after death. The feasibility of using DNA extracted from forensic bloodstain specimens for the detection of DNA polymorphisms was explored using a human haptoglobin (Hp) alpha chain specific probe. Using HindIII and XbaI digests the Hp genotypes Hp2, Hp1F and Hp1S were distinguished by Southern blot analysis in DNA prepared from 1 cm2 bloodstains up to 15-18 months old.

Autoradiography↗

Haptoglobin typing from fatty tissues after extraction with non-ionic detergents.

Of the seven non-ionic detergents tested, Span 40, Tween 20 and Triton X-45 were suitable for extraction of haptoglobin (Hp) from fatty tissues. The extracted samples were subjected to electrophoresis and subsequently stained with o-tolidine. Enzyme-immunoassay and Western blotting (electrophoretic) techniques were used successfully to determine the Hp phenotypes from minute quantities of tissue samples after extractions with non-ionic detergents.

Adipose Tissue↗

Tyrosine aminotransferase and chymotrypsinogen B are linked to haptoglobin on human chromosome 16q: comparison of genetic and physical distances.

The loci for haptoglobin (HP) and tyrosine aminotransferase (TAT) are known to reside at 16q22. Chymotrypsinogen B (CTRB), which is syntenic with TAT and HP on mouse chromosome 8, has also been assigned to human chromosome 16 but has not been mapped regionally. A linkage analysis was carried out in 13 informative families using RFLPs for these three markers. For CTRB, two TaqI RFLPs with a polymorphism information content of 0.60 derived from haplotype frequencies are described. The most likely order of loci, deduced from triple informative crosses, and their map distances, obtained by pair-wise linkage analysis, are HP-7 cM-TAT-9 cM-CTRB. By pulsed-field gel electrophoresis, a physical map covering more than 2000 kb was constructed. A maximum physical distance of about 700 kb was obtained for HP and TAT, which contrasts with the genetic distance of 7 cM (approximate confidence limits 2-18 cM). CTRB is at least 800 kb away from these two markers.

Amino Acid Metabolism, Inborn Errors↗

Acute phase-dependent changes in the binding of rat liver nucleoproteins to the cytokine response element of the rat haptoglobin gene.

Hormones released during the acute phase reaction promote the transcriptional activation of the haptoglobin (Hp) gene and a consequent increase of Hp protein synthesis in the liver. The mechanisms underlying the alterations of basal transcription rates of eukaryotic genes are assumed to result from modulations of the binding affinities between nucleoproteins and specific DNA sequences in the enhancer and promoter elements. In order to characterize the changes in the interaction of nucleoproteins with the promoter that accompany the induction of the Hp gene, nuclear extracts from normal and inflamed livers were probed with hormone responsive element (HRE) of the rat Hp gene by gel mobility shift and Southwestern assays. Each of the three cis-acting sequences of the HRE, elements A, B, and C, recognized a distinct set of proteins. Together they conferred an additional level of specificity to the protein binding sites of the entire ABC-element. These sites were recognized by proteins in liver nuclear extracts isolated from both control and treated rats. The differences in the gel shift and Southwestern patterns of the corresponding DNA-protein complexes suggested that transcriptional activation of the Hp gene relied on changes in the concentrations and/or functional modifications of preexisting proteins rather than on the induction of new trans-acting factors.

Acute-Phase Proteins↗

Dual inhibition of cyclooxygenase and lipoxygenase by human haptoglobin: its polymorphism and relation to hemoglobin binding.

Haptoglobin (Hp) binds hemoglobin (Hb) specifically and stoichiometrically. Since Hb stimulates prostaglandin (PG biosynthesis), we investigated if Hp effects arachidonic acid (AA) metabolism. The results showed that Hp (50-250 microg protein) inhibited the biosynthesis of PGs via cyclooxygenase (COX) and 12-HETE via lipoxygenase pathway in human platelets. Additional evidence was obtained by the loss of Hp inhibitory activity upon removal of Hp by affinity chromatography on hemoglobin sepharose and by inhibition of AA or bradykinin-induced bronchoconstriction in the guinea pig. Hb reduced the inhibitory effect of Hp in a concentration-related manner such that all its inhibitory activity was lost when completely bound by Hb. Of the three Hp phenotypes, Hp 1-1 showed maximum binding capacity to Hb indicating its greater protective role. These findings implicate Hp in the regulation of COX and lipoxygenase pathways and show Hp involvement in the body's endogenous defense system against inflammation. This indicates that mammals have dual defense system, i.e., a specific immune system and non-specific Hp defense system.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

Elevated plasma concentrations of haptoglobin in European brown bears during hibernation.

Haptoglobin (Hp), a hemoglobin-binding protein, is known as an acute phase protein and increases during the acute phase of inflammation in most mammals. We reported previously in brown bears that the mean Hp concentrations were higher in blood samples obtained in winter than those in spring. To examine a possible relation of the seasonal variations of Hp to hibernation, in the present study, we measured the plasma concentrations of Hp as well as some other acute phase proteins (alpha(2)-macroglobulin, alpha(1)-antitrypsin, C-reactive protein) in 6 European brown bears (Ursus arctos), from which blood samples were obtained at 5-6 different months of year including February, the time of hibernation. The Hp concentrations showed clear seasonal variations, being highest in February. The alpha(2)-macroglobulin concentrations also showed a similar but much smaller rise in February, but those of alpha(1)-antitrypsin and C-reactive protein did not show any seasonal variations. Our results suggest that the seasonal variation of plasma Hp concentration in brown bears is associated with a hibernation-specific mechanism more than that of acute phase response.

Animals↗

Acute-phase related binding ability of p53 for the hormone response element of the haptoglobin gene in adult rats.

Interaction between transcription factor p53 and the hormone response element (HRE) of the haptoglobin (Hp) gene in adult rat liver was studied. We detected a sequence homologous to the p53 consensus DNA-binding site in the regulatory promoter element of the Hp gene. DNA-affinity chromatography, followed by Western immunoblot analysis with an antibody to p53 indicated that components of the nuclear extract possessed the same antigen determinants as p53. While p53 was identified in both control and acute-phase (AP) samples, DNA-binding affinity for the Hp gene HRE was detected only in the nuclear extract prepared from rats undergoing the AP response. Whether either as an inducible or as a constitutive transcription factor, p53 could be involved in the transcriptional regulation of the Hp gene in adult rat liver.

Acute-Phase Reaction↗

Temporal changes in concentrations of serum amyloid-A and haptoglobin and their associations with weight gain in neonatal reindeer calves.

Age-related changes in serum concentrations of two acute phase proteins (APPs), haptoglobin (Hp) and serum amyloid-A (SAA) were investigated in newborn reindeer calves. Repeated blood samples were obtained from 51 reindeer calves at ages 0-32 days (2-4 samples from each calf). An increase of SAA concentrations was observed during the first 2 weeks of life. However, by the end of the observation period, SAA concentrations had decreased to levels below those of the first week. Serum Hp concentrations increased throughout the observation period. SAA concentrations in the second week had a negative association with weight gain during the entire study period (4 months). These time-related changes in APP concentrations suggest that these proteins have a role in the defence and adaptation mechanisms of newborn reindeer calves. Possible reasons for these changes include the presence of APP mediators in the colostrum, exposure to environmental pathogens after birth and age-related changes in hepatic synthesis of APP.

Age Factors↗