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Decreases of apolipoprotein B-100 and A-I concentrations and induction of haptoglobin and serum amyloid A in nonfed calves.

Reduced feed intake near parturition is suggested to be one of the major causal factors for the development of fatty liver in cows, and nonfeeding has been used as an experimental model for fatty liver. In cows with fatty liver, concentrations of lipoprotein lipids and proteins are decreased. In addition, the acute-phase protein haptoglobin is induced. The purpose of the present study was to examine whether the decrease of lipoprotein concentrations and the induction of acute-phase proteins were similarly reproduced by non-feeding. Holstein female calves (n=5) were nonfed for 3 days and thereafter refed. Serum concentrations of nonesterified fatty acids and beta-hydroxybutyric acid were initially increased by the nonfeeding, and followed by decreases in concentrations of cholesteryl esters, phospholipids, apolipoprotein (apo) B-100 and apoA-I. The apoC-III concentration was not distinctly decreased. Haptoglobin and serum amyloid A were induced during the nonfeeding and refeeding process. Haptoglobin was distributed in different proportions in the high-density lipoprotein, very high-density lipoprotein and the lipoprotein-deficient fractions, whereas almost all serum amyloid A was associated with the high-density lipoprotein fraction. These results suggest that the decreases in lipoprotein concentrations and induction of acute-phase proteins found in cows with fatty liver and those with fatty liver-related diseases such as ketosis are primarily due to the reduced feed intake near parturition.

3-Hydroxybutyric Acid↗

Dynamics in serum of the inflammatory markers serum amyloid A (SAA), haptoglobin, fibrinogen and alpha2-globulins during induced noninfectious arthritis in the horse.

Despite the importance of noninfectious joint diseases in equine medicine, little is known about the acute phase response which may be elicited if the local inflammatory process of noninfectious arthritis is sufficiently strong, Therefore the aim of this study was to monitor the systemic inflammatory response during experimentally-induced noninfectious arthritis by studying the dynamics in serum of the acute phase proteins serum amyloid A (SAA), haptoglobin, fibrinogen and alpha2-globulins. Twenty-four Standardbred horses, age 3-7 years, found healthy on thorough clinical, radiological, haematological and serum biochemical examination, were injected aseptically into the right midcarpal joint with amphotericin B. Blood samples were drawn before induction of arthritis (0 h), and at 8, 16, 24, 36 and 48 h postinduction and then on Days 3, 4, 5 and 15 postinduction. All horses developed lameness with joint effusion and joint heat as well as increased respiratory rate, heart rate and body temperature. The lameness started to decline after 24-36 h and, in most animals, systemic signs disappeared on Day 2 postinjection. The concentration of the acute phase proteins increased following induction of arthritis. The SAA concentrations were higher than baseline concentrations from 16 h postinduction and were maximal at 36-48 h (227 times baseline concentration). The haptoglobin concentrations were higher than baseline concentrations from 24 h and were maximal at 48-96 h (1.14 times baseline concentration). The maximal concentrations of fibrinogen were seen between 36-72 h postinjection and increased on average 0.87 times from baseline concentrations. The fibrinogen concentrations were higher than baseline concentrations from 24 h postinjection. Alpha2-globulins concentrations showed a minor increase and increased 0.55 times from baseline concentrations. The markers had returned to baseline concentrations by Day 15. Our results demonstrate that amphotericin B-induced arthritis in a single joint gives rise to a systemic acute phase response measurable as increased concentrations in serum SAA, haptoglobin, fibrinogen and alpha2-globulins during the first 2 weeks of the condition and, thereby, that such an increase need not be indicative of infectious arthritis. Further research should be aimed at determining whether chronic noninfectious arthritis in the horse gives rise to increased acute phase protein concentrations in serum.

Acute-Phase Proteins↗

A simple and rapid method for purification of rat haptoglobin for production of antiserum.

Plasma from rats with acute inflammatory response was fractionated on Blue Sepharose CL 6B, to separate haptoglobin from albumin and lipoproteins. Affinity chromatography on Blue Sepharose proved to be a convenient method for crude fractionation of plasma. Pure haptoglobin was obtained by the subsequent affinity chromatography on a rabbit-haemoglobin Sepharose column. Minor amounts of rabbit haemoglobin co-eluted from the haemoglobin Sepharose column but did not influence the monospecificity of the antiserum raised in rabbits. By use of the antiserum in single radial immunodiffusion, the concentration of haptoglobin in plasma from normal rats was measured to be 0.5 g/l.

Acute-Phase Reaction↗

[Behavior of prealbumin, retinol-binding protein, transferrin and haptoglobin in the postoperative and post-traumatic phase].

The plasma proteins pre-albumin, retinol-binding protein, transferrin, and haptoglobin were determined in 3 different groups of surgical patients (8 patients after inguinal or incisional hernia operation, 12 patients with peritonitis due to insufficient anastomoses, 17 patients with skull-brain injuries) according to the radial immune diffusion method. The decrease in concentration of nutrition-dependent plasma proteins, pre-albumin, retinol-binding protein and transferrin, varied depending on the extent of postoperative or posttraumatic catabolism and respective nutritional regimen. The increase in concentration of the acute phase protein haptoglobin was not significantly dependent on the severity of trauma. There was, however, a further haptoglobin increase of significance due to postoperative peritonitis complication.

Haptoglobins↗

Detection of annexin I and IV and haptoglobin in bronchoalveolar lavage fluid from calves experimentally inoculated with Pasteurella haemolytica.

OBJECTIVE: To determine whether annexins or haptoglobin could be detected in bronchoalveolar lavage (BAL) fluid specimens obtained from calves experimentally inoculated with Pasteurella haemolytica. ANIMALS: Twelve 2- to 3-month-old male Holstein calves. PROCEDURE: Pasteurella haemolytica was inoculated into the right lung lobes of each of 6 calves. Six other calves received vehicle alone and were used as control calves. Specimens of BAL fluid were obtained from 3 control and 3 inoculated calves 1 day after inoculation and from the other calves 2 days after inoculation. The amount of annexins I, II, IV, and VI, and haptoglobin in BAL fluid specimens was examined by use of immunoblot analysis. RESULTS: Annexins I and IV were detected in BAL fluid specimens obtained from the right lung lobes of each of the inoculated calves, but annexins II and VI were not. Annexin I also was found in BAL fluid specimens obtained from the left lung lobes of each inoculated calf and from left and right lung lobes of the control calves. By comparison, detection of annexin IV was essentially limited to the right lung lobes of inoculated calves. Haptoglobin was detected in some, but not all, BAL fluid specimens from the right lung lobes of inoculated calves, and its detection in BAL fluid was associated with serum proteins such as albumin. CONCLUSIONS AND CLINICAL RELEVANCE: Annexin IV was detected most specifically in response to inoculation of P haemolytica. This protein could be used as a marker for inflammatory pulmonary disease caused by P haemolytica.

Animals↗

Simple PCR detection of haptoglobin gene deletion in anhaptoglobinemic patients with antihaptoglobin antibody that causes anaphylactic transfusion reactions.

Two anhaptoglobinemic patients showing anaphylactic transfusion reactions by antihaptoglobin antibody were found. Southern blot analysis indicated that 2 patients were homozygous for the deleted allele of the haptoglobin gene (Hp(del)) as reported previously. We have identified the junction region of the deletion from genomic DNA of 1 patient using cassette-mediated polymerase chain reaction (PCR). Then, the deleted region from the 5' breakpoint to the promoter region of the Hp was amplified from genomic DNA of a control individual using PCR. DNA sequence analysis of these regions indicated that the 5' breakpoint of the Hp(del) allele was located 5. 2 kilobase (kb) upstream of exon 1 of the Hp and the 3' breakpoint was positioned between 52 and 53 base pair (bp) upstream of exon 5 of the haptoglobin-related gene. There was no significant homology between the DNA sequences flanking the 5' and 3' breakpoints, except for a 2-bp (TG) identity. To examine the gene frequency, we have developed a simple PCR method to detect the gene deletion. We found 8, 16, and 17 Hp(del) alleles in 157 Koreans, 523 Japanese, and in 284 Chinese, respectively, but did not find the Hp(del) in 101 Africans or in 100 European-Africans. The incidence of individuals homozygous for the Hp(del) allele was therefore expected to be 1/4000 in Japanese, 1/1500 in Koreans, and 1/1000 in Chinese. This incidence is higher than that of IgA deficiency in Japanese. More attention should be paid on haptoglobin deficiency and antihaptoglobin antibody as the cause of transfusion-related anaphylactic reactions in Asian populations. (Blood. 2000;95:1138-1143)

5' Untranslated Regions↗

Haptoglobin: function and polymorphism.

Haptoglobin is an acute phase protein capable of binding haemoglobin, thus preventing iron loss and renal damage. Haptoglobin also acts as an antioxidant, has antibacterial activity and plays a role in modulating many aspects of the acute phase response. There are 3 major haptoglobin phenotypes--Hp(1-1), Hp(2-1) and Hp(2-2). Possession of a particular phenotype has been associated with a variety of common disorders (e.g. cardiovascular disease, autoimmune disorders, malignancy), a fact which can only be explained by the idea that possession of a particular phenotype offers some protection against the development of these disorders. Knowledge of phenotype could therefore aid in the prognosis of disease and allow treatment to be better tailored to suit an individuals' needs.

Autoimmune Diseases↗

[Features of disruption of certain components of carbohydrate metabolism in a combination of pulmonary tuberculosis and diabetes mellitus in people with haptoglobin phenotypes].

Sugar level in blood, the activity of lactate dehydrogenase (LDH), glucose-6-phosphate dehydrogenase (G-6-PDH), 2,3-BPG content, HbA1C and the phenotype of haptoglobin were studied in 180 patients with lung tuberculosis and diabetes mellitus. The increased (2-4.2-fold) blood sugar level was found in 77.2% patients. It was accompanied by decreased activity of LDH (by 1.3-1.7 times), G-6-PDH (by 15-45% in 87% patients). In patients with various haptoglobin phenotypes the content of HbA1C and 2.3-BPG was increased by 1.5-1.7 and 2-3 times, respectively. Clear differences in the studied parameters were found in patients with various phenotypes of haptoglobin (Hp). The most serious impairments of the studied parameters of carbohydrate metabolism were found in untreated patients with homozygote Hp phenotypes 2-2 and 1-1. Alterations found in the present study can be used for evaluating the depth of impairments of the carbohydrate metabolism in patients with combination of lung tuberculosis and diabetes mellitus.

Carbohydrate Metabolism↗

The application of immunoblotting to the phenotyping of haptoglobin.

An immunoblotting method for phenotyping haptoglobin in serum and bloodstains has been developed. Haptoglobin isoproteins were separated by polyacrylamide gradient gel electrophoresis and then transferred to nitrocellulose by electroblotting. The use of 1 mm gels facilitated more rapid and effective transfer than conventional 3 mm thick gels. Nitrocellulose blots were developed by double antibody enzyme immunoassay. The detection limit for serum and bloodstains was improved 16 times compared to conventional staining using O-tolidine. The method could detect haptoglobin phenotypes from 0.001 microliter of whole blood. This detection limit is approximately 8 times lower than that of group specific-component analysis by immunoblotting.

Animals↗

Serum haptoglobin concentration as an indicator of weight gain in pigs.

Serum haptoglobin concentration was investigated as an indicator of weight gain in commercially-reared pigs. The serum haptoglobin concentrations and weights of 40 pigs were monitored on a weekly basis, from weaning to 13 weeks of age. All data were ranked based on the week 13 weights, and divided into high, middle and low weight gain groups. By the fourth week of the study, when the pigs were seven weeks old, serum haptoglobin concentrations could be used to differentiate pigs that would have a high weight gain at the end of the study from those which would have a low weight gain.

Analysis of Variance↗

Pretreatment serum levels of C-reactive protein, alpha 1-antitrypsin, haptoglobin, alpha 1-acid glycoprotein and tissue polypeptide antigen in cervical carcinoma.

In order to evaluate the potentially additive information of some acute phase reactants to that provided by a general tumour marker, pretreatment concentrations of C-reactive protein, alpha 1-antitrypsin, haptoglobin, alpha 1-acid glycoprotein and tissue polypeptide antigen were determined in serum from healthy women, patients with dysplasia/or carcinoma in situ and patients with primary cervical carcinoma. Specificity varied from 95-100% and sensitivity from 16-29%. A correlation with clinical stage was found for all analytes except for alpha 1-antitrypsin. The latter was the most frequently elevated analyte in early Stages (11/43 in Stage Ib/IIa) and uniquely elevated in 7 cancer patients. Although tissue polypeptide antigen predominantly signaled in advanced stages, 3 women in early stages had elevated tissue polypeptide antigen levels. One of these women died and she was also the only woman with raised alpha 1-antitrypsin who died. It is discussed whether elevated tissue polypeptide levels might represent an unfavourable sign for the individual and if alpha 1-antitrypsin is a favourable sign in early stages of cervical carcinoma. C-reactive protein results were obscured in early stages of disease by the presence of intercurrent illness and the results were regarded as inconclusive. Haptoglobin and alpha 1-acid glycoprotein concentrations provided no additional information to serum alpha 1-antitrypsin levels. However, haptoglobin was elevated in 64% (36/56) of the women with dysplasia/carcinoma in situ of the cervix uteri.

Acute-Phase Proteins↗

[Effect of ACTH on the excretion of acid glycosaminoglycans in urine and on the content of haptoglobin in blood serum of rats in acute aseptic inflammation].

In aseptic inflammation an increase in excretion of acid glucosoaminoglycans with urine of rats was an earlier evidence for destructive processes in the main substance of connective tissues than an increase in content of haptoglobin in blood serum. The increase in content of haptoglobin in blood serum was probably caused by beginning reparative processes in the centre of impairment. The centre of inflammation, caused by subcutaneous administration of turpentine simultaneously with repeated injections of high doses of ACTH, was maintained in the rats longer than in animals, which were not treated with ACTH. In the case the most distinct increase in excretion of glycosoaminoglycans with urine was observed; content of haptoglobin was also increased in blood serum.

Acute Disease↗

Haptoglobin typing in canine bloods.

Haptoglobin typing by vertical electrophoresis in a discontinuous polyacrylamide gel was conducted on 47 dog blood samples, of which 19 were from Doberman pinschers, 20 from German shepherds, and 8 from pit bullterriers. Two phenotypes were common in the three breeds and could not be used to differentiate between them. Canine haptoglobin phenotypes were, however, sufficiently different from those of humans to warrant using haptoglobin typing as a method for determining the origin of bloodstains.

Animals↗

[The effect of haptoglobin on prostaglandin H synthase from ram vesicular glands].

It was shown that the ability of sheep and horse haptoglobins differing in their immunological properties to inhibit PGH synthetase is about the same. It was found that haptoglobin inhibits the PGH synthetase-catalyzed enzymatic reaction, the inhibiting effect being non-competitive with respect to the electron donor, adrenaline. The degree of PGH synthetase inhibition by haptoglobin depends on the glycoprotein concentration, incubation time and enzyme activity.

Animals↗

Serum haptoglobin level in patients with malignant lymphoma and its clinical significance.

Serum haptoglobin level and its phenotypes were studied in 208 normal people and 176 malignant lymphoma patients consisting of 112 Non-Hodgkin's Lymphoma (NHL) and 64 Hodgkin's Disease (HD) at Tata Memorial Hospital, Bombay and were correlated with the clinical stages of the lymphomas. Haptoglobin (Hp) levels were also estimated in the patients at different intervals during their chemotherapy. Hp level was found significantly increased to 147.83 +/- 8.67 in NHL and 173.57 +/- 13.31 in HD as compared to 67.46 +/- 2.53 in the controls. The phenotypic and gene frequencies in the lymphoma patients did not differ significantly from those of the controls. All the clinical stages, except stage I, of NHL and HD revealed significant rise in Hp level when compared with the controls. When different clinical stages of NHL and HD were compared separately, HD showed significant rise in HP level over the NHL only in Stage IV (p less than 0.05). Significant reduction in Hp level was observed when the disease regressed due to chemotherapy but it again increased with the relapse of the disease. Thus, the study of serum haptoglobin levels at intervals in lymphoma patients would help to understand the regression or relapse of the disease and also the prognosis.

Haptoglobins↗

Haptoglobin and proteinase inhibitors in the blood serum of women with inflammatory, benign and neoplastic lesions of the ovary.

Blood sera of women with myomas of the uterus, benign or malignant neoplasms of the ovary and with inflammatory conditions were examined by determining the haptoglobin concentration and functional activities of: alpha 2-macroglobulin, alpha 1-proteinase inhibitor and alpha-cysteine proteinase inhibitor. In the women with ovarian cancer, as compared with the control group, a significant increase of haptoglobin, alpha 1-proteinase inhibitor and alpha-cysteine proteinase inhibitor was found (p less than 0.001). On the contrary bioactivity of alpha 2-macroglobulin was depressed (p = 0.01). The observed increase of alpha-cysteine proteinase inhibitor activity in the groups with ovarian tumors and ovarian cysts was not related to the inflammatory state as estimated by the haptoglobin concentration.

Adult↗

Evaluation of serum haptoglobin levels in patients with adnexal masses.

Over a four-year period, serum haptoglobin was measured in 152 patients with adnexal masses in an attempt to discriminate between benign and malignant disease. When serum haptoglobin exceeded 150 mg/dl, malignancy could be identified with a sensitivity of 83% and a specificity of 84%. While the predictive value of a positive test is only 66%, the predictive value of a negative test is 93%, and this inexpensive test may be clinically useful. Stage, grade, and size of tumor did not correlate with serum haptoglobin levels.

Adnexa Uteri↗

A population genetic survey of the haptoglobin polymorphism in Melanesians by DNA analysis.

We have determined the haptoglobin (Hp) genotypes of 831 Melanesians from Vanuatu, Papua New Guinea, and New Caledonia by Southern blot analysis of DNA extracted from umbilical cord and peripheral blood samples. There was complete agreement between these genotypes and the protein phenotype in cases where both were determined, and genotyping was possible in cases where no serum haptoglobins were measurable. Subtyping of Hp1 alleles using restriction enzymes showed that Melanesians, like Mongoloids and Australian Aboriginals, have only the Hp1S allele. Three cases of Hp Johnson were found in Vanuatu, and further restriction mapping supported a partial gene triplication model for the structure of this variant. We also report a new common BclI restriction enzyme polymorphism upstream of the Hp1 gene. The advantages of using DNA for haptoglobin typing are discussed.

Alleles↗