The cDNA encoding inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP) was cloned from human liver cDNA libraries. Oligonucleotide primers of human liver cDNA for PCR were constructed from internal amino acid sequences obtained with proteolytic fragments of IHRP. The amplified cDNA served as a hybridization probe for the screening of human liver cDNA libraries. The cDNA of 2,977 bp contained an entire reading frame coding 930 amino acids. The N-terminal 28 residues corresponded to a signal peptide for secretion. The N-terminal 600 residues of the mature form exhibited considerable homology to those of ITI heavy chains, while the C-terminal 300 residues showed no homology with the heavy chains and low homology with ATP-dependent proteases. IHRP was readily cleaved into 85- and 35-kDa fragments when plasma was incubated at 37 degrees C. The cleaved site, Arg-Arg-Leu, was within a proline-rich region. Northern blot analysis of poly(A) RNAs from various human tissues only showed hybridization to liver RNA.

OBJECTIVE

In the present study, the involvement of the binding of IHRP (inter-alpha-trypsin inhibitor family heavy chain-related protein) and actin in phagocyte activity was investigated.

METHODS

The actin polymerization and the phagocytic activity of the polymorphonuclear (PMN) cells were studied in the presence of IHRP.

RESULTS

IHRP inhibited the polymerization of actin and the phagocytic activity of the PMN cells.

CONCLUSIONS

1) IHRP may bind to actin released from the damaged cells and suppress its toxic action by preventing the formation of actin fibril. 2) IHRP may bind to cell surface actin on PMN cells and inhibit their phagocytic activities. 3) From these results, IHRP may act as an anti-inflammatory protein.

Inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP), which has a sequence similarity to the heavy chains of the inter-alpha-trypsin inhibitor (ITI) family, is a novel glycoprotein found in human plasma. We prepared two clones (1A4 and 6E11) of anti-IHRP mouse monoclonal antibody. Both of them recognized the C-terminal 35-kDa fragment which was produced by plasma kallikrein-digestion of IHRP. We developed a sandwich ELISA for measurement of the plasma IHRP concentration with the monoclonal antibody coated microtiter plate and the anti-N-terminal 57-kDa fragment of IHRP rabbit polyclonal antibody (anti-GP57). We found that the average concentration of IHRP in the plasma of healthy donors was 101.3+/-31.8 microg/ml (average+/-S.D.). The IHRP concentration in the plasma of patients with inflammatory disorders was slightly increased (137.5+/-40.2 microg/ml: average+/-S.D.). Together with the previous data indicating the induction of the porcine mRNA, which is thought to be the species counterpart of human IHRP mRNA, in the liver after resuscitation from cardiogenic shock, we propose that IHRP is a member of the acute-phase protein family.

Inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP) is a novel glycoprotein isolated from human plasma. The cDNA encoding IHRP has recently been cloned from human liver cDNA libraries. We report the mapping of this gene (ITIHL1) by fluorescence in situ hybridization using a 2.5-kb cDNA fragment as a probe. ITIHL1 was localized to chromosome region 3p21->>p14 where the genes of heavy chain 1 and 3 of inter-alpha-trypsin inhibitor are located. This result, together with significant homology between the nucleotide sequences of ITIHL1 and the heavy chain genes, supports ITIHL1 as being a member of an evolutionary related gene family of ITI heavy chains. Northern blot analysis indicated that IHRP was predominantly synthesized in liver. From Southern blot analysis, it was tentatively concluded that ITIHL1 is a single copy gene.

Plasmapheresis with a dextran sulfate column is a treatment for patients with hypercholesteremia. When proteins bound to the column during the treatment were fractionated to prepare some known proteins, we found a 57 kDa glycoprotein designated GP57 which showed a new N-terminal amino acid sequence. Western-blot analysis of human plasma revealed that only a 120 kDa protein, GP120, reacted with anti-GP57 antibody. Since GP120 and GP57 had an identical N-terminal amino acid sequence, GP120 is probably the intact form of GP57. The isoelectric point of GP120 was 6.8. N-Glycanase treatment decreased the molecular weight of GP120 by 15 kDa. Neuraminidase and O-glycanase, however, did not affect the molecular weight. Amino acid sequence analyses of the lysylendopeptidase digest of GP120 revealed significant homology to the heavy chains of inter-alpha-trypsin inhibitor (ITI) family. Since GP120 showed no bikunin sequence, and chondroitinase treatment and alkaline treatment of GP120 did not affect its molecular weight, we concluded that GP120 was not a complex with bikunin. We designated GP120 as IHRP (ITI heavy chain-related protein).

A novel human plasma protein was found in the eluate from the dextran sulfate column, which was used for the treatment of the patients with hypercholesteremia to reduce plasma low density lipoprotein. The results of sequence analysis revealed that this protein was a homologue of heavy chains of inter-alpha-trypsin inhibitor (ITI) family, and it was termed IHRP (ITI family heavy chain related protein). IHRP was identified as an acute-phase protein in animals, and slightly increased concentrations in human plasmas were observed in the patients with inflammatory disorders. IHRP bound to actin and inhibited its polymerization, and IHRP suppressed the phagocytosis and chemotaxis of polymorphonuclear cells. These results suggest that IHRP may function as an anti-inflammatory protein. Plasma hyaluronan binding protein (PHBP) is a novel serine protease which was also found in human plasma. It is consisted of three epidermal growth factor domains, one kringle domain and one serine protease domain from its amino terminus. The amino acid sequence of PHBP is homologous to that of hepatocyte growth factor activator. Purified 75-kDa single chain pro-form of PHBP was auto-activated (auto-cleaved) to 50-kDa heavy chain and 25-kDa light chain, both of them are bridged by a disulfied bond. PHBP digested alpha-chain and beta-chain of fibrinogen to prevent coagulation and cleaved single chain urokinase type plasminogen activator (scuPA) to the active hetero dimer form (tcuPA). The auto-activation of PHBP was accelerated in the presence of dextran sulfate or phosphatidylethanolamine as well as factor XII of the coagulation system. C1 inhibitor of the complement system was identified as the main inhibitor of PHBP in human plasma. Partial hepatectomy and administration of carbon tetrachloride or galactosamine caused the conversion of pro-PHBP to the active form in mouse but administrations of turpentine and mercury chloride did not, suggesting the hepatic injury specific activation of PHBP. These results indicate that PHBP participates not only in the fibrinolytic system but also in the degradation cascade of extracellular matrix (ECM), i.e., PHBP activates scuPA to tcuPA, tcuPA activates matrix metalloproteases (MMPs) and activated MMPs degrade ECM for the tissue remodeling after hepatic injury.

The pig counterpart of human IHRP has been isolated from pig serum, and cDNA clones encoding this counterpart were isolated and characterized. The amino acid sequence of pig IHRP predicted from the nucleotide sequence of its cDNA shows reasonable homology to that of human IHRP. The nucleotide sequence of pig IHRP cDNA is identical to that of the mRNA partially determined to be the heavy chain of pig ITI [Buchman et al. (1990) Surgery 108, 560-566]; it is one of the major mRNAs induced in pig liver on cardiogenic shock. Thus we concluded that the reported mRNA should code for IHRP and not for the heavy chain of ITI. The pig IHRP also seems to be identical to pig-MAP, which was recently reported to be a major acute phase serum protein in pig [Gonzalez-Roman et al. (1995) FEBS Lett. 371, 227-230]. The results suggest that IHRP might be involved in acute phase reactions.

BACKGROUND

Ash tree (Fraxinus excelsior) is the main representative of the Oleaceae family in temperate zones. Diagnosis of ash pollen allergy is made difficult due to (1) an overlapping pollinization period with Betulaceae, (2) non-inclusion in current diagnostic assays, and (3) some cross- reactivity with minor allergens from Betulaceae. The aim of this study was to calibrate an ash pollen in-house reference preparation (IHRP) in allergic patients in order to produce standardized products for diagnosis and immunotherapy purposes.

METHODS

Ash pollen IHRP was extracted, ultrafiltered and freeze dried. Allergens in the extract were detected after 2-dimensional PAGE using specific sera and a monoclonal antibody. The Fra e 1 content of IHRP was evaluated by quantitative immunoprint. Forty-eight subjects from the North-East of France exhibiting clinical symptoms, a positive skin test and specific IgE levels>> or =class 2 to ash pollen were recruited. IgE immunoprints were performed to select patients sensitized to the ash Fra e 1 allergen as opposed to cross-reacting allergens. Serial 10-fold dilutions of the IHRP were tested by skin prick tests in order to determine the concentration inducing a geometrical mean wheal diameter of 7 mm, said to correspond to an index of reactivity (IR) of 100 per millilitre.

RESULTS

IgE-reactive molecules in IHRP comprise Fra e 1, Fra e 2, a 9-kDa molecule (presumably Fra e 3), as well as a doublet at 15 kDa and high molecular weight allergens. The 100 IR concentration of IHRP inducing a geometrical mean wheal diameter of 7 mm in 22 patients sensitized to Fra e 1 corresponds to the 1/126 (w/v) extraction ratio (i.e. 259 microg/ml of protein by Bradford) and contains 17 microg/ml of Fra e 1. The variability in total activity of 5 batches of standardized extracts was found to be significantly reduced when compared with 7 non-standardized extracts.

CONCLUSIONS

An ash pollen IHRP was defined and molecularly characterized. Its successful standardization at 100 IR/ml in patients specifically sensitized to Fra e 1 allowed a skin reactivity-based calibration in properly diagnosed patients. Such a standardized ash pollen extract is a reliable tool to support immunotherapy of ash pollen allergy.

Inter-alpha-trypsin inhibitor (ITI) family heavy chain-related protein (IHRP) is a novel human glycoprotein that shows significant homology in amino acid sequence to proteins of the ITI family heavy chains from human plasma. Three overlapping clones that encode the human inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP) gene (ITIHL1) were isolated and characterized. The IHRP gene spans 15 kb and is composed of 24 exons from 27 to 207 bp in size with consensus splice sites. The gene codes for the precursor of IHRP, which is similar to inter-alpha-trypsin inhibitor (ITI) family heavy chains. Two major transcription initiation sites were identified in the 5'-flanking region. They contain putative promoter elements, but no typical TATA box. Some exons of this gene showed significant similarities to those of the ITI-H1 gene in nucleotide length and in intron phasing. The tissue-specific transcription of this gene may be due to the presence of binding sites for the hepatocyte nuclear factors LF-A1, HNF-5, NF-IL6, and C/EBP. This gene was found to be localized very close to another unknown gene related to EST (GenBank accession #: R54643, R50663, R50563, H27139, and R54913).

In this paper, we give a systematic study to report several deep insights into the HOG, one of the most widely used features in the modern computer vision and image processing applications. We first show that, its magnitudes of gradient can be randomly projected with random matrix. To handle over-fitting, an integral histogram based on the differences of randomly selected blocks is proposed. The experiments show that both the random projection and integral histogram outperform the HOG feature obviously. Finally, the two ideas are combined into a new descriptor termed IHRP, which outperforms the HOG feature with less dimensions and higher speed.

1. In this study, electrocardiographic second lead tracings were recorded and analyzed online and offline in the conscious and spinalized Xenopus laevis under control conditions and after treatment with at least two effective doses of racemic diltiazem, nifedipine and racemic verapamil hydrochlorides. 2. Heart rates (HR) and intercycle heart rate power spectra (IHRPS) were calculated in the various experimental conditions. 3. Dose-dependent bradycardia was found in most cases, which was associated with dose-related increases in the overall dynamic range of the largest peak of HR variability (increased IHRPS), this being particularly marked at the higher nifedipine dose. The frequency position of the largest peak was unchanged in the spinalized preparation, as well as in the various treatments. 4. These findings may contribute toward a more complete pharmacodynamic definition of calcium channel blockers in their current therapeutic use.

Leprosy is a chronic dermato-neurological disease caused by Mycobacterium leprae infection. In 2016, more than 200,000 new cases of leprosy were detected around the world, representing the most frequent cause of infectious irreversible deformities and disabilities.

In the present work, we demonstrate a consistent procoagulant profile on 40 reactional and non-reactional multibacillary leprosy patients. A retrospective analysis in search of signs of coagulation abnormalities among 638 leprosy patients identified 35 leprosy patients (5.48%) which displayed a characteristic lipid-like clot formed between blood clot and serum during serum harvesting, herein named 'leprosum clot'. Most of these patients (n = 16, 45.7%) belonged to the lepromatous leprosy pole of the disease. In addition, formation of the leprosum clot was directly correlated with increased plasma levels of soluble tissue factor and von Willebrand factor. High performance thin layer chromatography demonstrated a high content of neutral lipids in the leprosum clot, and proteomic analysis demonstrated that the leprosum clot presented in these patients is highly enriched in fibrin. Remarkably, differential 2D-proteomics analysis between leprosum clots and control clots identified two proteins present only in leprosy patients clots: complement component 3 and 4 and inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP). In agreement with those observations we demonstrated that M. leprae induces hepatocytes release of IHRP in vitro.

We demonstrated that leprosy MB patients develop a procoagulant status due to high levels of plasmatic fibrinogen, anti-cardiolipin antibodies, von Willebrand factor and soluble tissue factor. We propose that some of these components, fibrinogen for example, presents potential as predictive biomarkers of leprosy reactions, generating tools for earlier diagnosis and treatment of these events.

A general monograph for allergen products has now been adopted by the European Pharmacopoeia Commission. It is based on the use of an In-House Reference Preparation (IHRP), which shall be used in the control of every production batch after appropriate characterization. This characterization will depend on the intended use of the product. When possible, the biological potency should be established by skin testing and expressed in biological units. The monograph is the first mandatory regulation in Europe for allergen products and will guarantee the consistent composition and potency of the products on the market.

This article analyzes intersectorial health-related policies (IHRP) based on a case study performed in 2008-2009 that mapped the social policies of the city of Piracicaba, State of Sao Paulo, Brazil. The research strategy comprised quantitative and qualitative methodologies and converging information sources. Legal and theoretical conceptual frameworks were applied to the Piracicaba study results and served as the basis for proposing a typology of IHRP. Three types of IHRP were identified: health policies where the health sector is coordinator but needs non-health sectors to succeed; policies with a sector other than health as coordinator, but which needs health sector collaboration to succeed; and thirdly, genuine intersectorial policies, not led by any one sector but by a specifically-appointed intersectorial coordinator. The authors contend that political commitment of local authorities alone may not be enough to promote efficient intersectorial social policies. Comprehension of different types of IHRP and their interface mechanisms may contribute to greater efficiency and coverage of social policies that affect health equity and its social determinants positively. In the final analysis,, this will lead to more equitable health outcomes.

To study the age-related changes of the sinus node function and the variations of influence of autonomic nervous system, pharmacologic total autonomic blockade (TAB) was conducted in 35 patients with symptomatic sinus bradycardia (21 men and 14 women, 50 +/- 21 years, mean +/- SD). Twenty-one patients [Group I, consisting of 14 patients younger than 60 years (group IY) nd 7 patients 60 years or older (group IE)] had a normal intrinsic heart rate (IHRo), and 14 patients had an abnormal IHRo [Group II, consisting of 8 patients younger than 60 years (group IIY) and 5 patients 60 years or older (group IIE)]. The basic cycle length was significantly longer in group II than in group I, suggesting that intrinsic sinus node function was more seriously deteriorated in group II in spite of the compensatory effect of autonomic regulation. In group II it was characteristic that the cycle length (CL) after atropine sulfate administration was longer than the CL of the predicted intrinsic heart rate (IHRp). Otherwise, some group II patients might be regarded as normal by atropine sulfate administration alone. Parasympathetic tone showed a negative correlation with age, and it was most enhanced in group IIY, suggesting that parasympathetic negative chronotropy was stronger in this group. In the course of propranolol administration, prolongation of CL was significantly larger in group IIE than in group IIY, but there was no age-related difference in group I. In group II, beta-adrenergic blockade with propranolol administration showed that sympathetic positive chronotropy was a critical compensatory factor around the upper limit of the CL of IHRp.(ABSTRACT TRUNCATED AT 250 WORDS)

BACKGROUND

Single-nucleotide polymorphisms (SNPs) are considered to be useful polymorphic markers for genetic studies of polygenic traits. Single-stranded conformational polymorphism (SSCP) analysis has been widely applied to detect SNPs, including point mutations in cancer and congenital diseases. In this study, we describe an application of the fluorescent labeling of PCR fragments using a fluorescent-adapted primer for SSCP analysis as a novel method.

METHODS

Single-nucleotide polymorphisms (SNPs) of the inter-alpha-trypsin inhibitor family heavy chain-related protein (IHRP) gene were analyzed using a fluorescence-adapted SSCP method. The method was constructed from two procedures: 1) a fluorescent labeling reaction of PCR fragments using fluorescence-adapted primers in a single tube, and 2) electrophoresis on a non-denaturing polyacrylamide gel.

RESULTS

This method was more economical and convenient than the single-stranded conformational polymorphism (SSCP) methods previously reported in the detection of the labeled fragments obtained. In this study, eight SNPs of the IHRP gene were detected by the fluorescence-adapted SSCP. One of the SNPs was a new SNP resulting in an amino acid substitution, while the other SNPs have already been reported in the public databases. Six SNPs of the IHRP were associated with two haplotypes.

CONCLUSIONS

The fluorescence-adapted SSCP was useful for detecting and genotyping SNPs.