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1.
Clin Radiol ; 73(11): 984.e11-984.e18, 2018 11.
Article in English | MEDLINE | ID: mdl-30072032

ABSTRACT

AIM: To estimate the morphological changes in the articular cartilage of the knees of patients with rheumatoid arthritis treated with biological disease-modifying anti-rheumatic drugs (bDMARDs). MATERIALS AND METHODS: Cartilage-specific magnetic resonance imaging (MRI) results, including T2 and T1ρ mapping of the femorotibial joint of 17 patients, were obtained before and 1 year after starting treatment with bDMARDs. Regions of interest were selected on the sagittal images of the cartilage of the medial and lateral femoral condyles (MFC, LFC) and the tibial plateau (MTP, LTP). Cartilage thickness, T2, and T1ρ were measured, and the correlations of their changes were evaluated. RESULTS: The mean changes in cartilage thickness tended to decrease in all four condyles, and the rate was significant in the MFC. T2 and T1ρ tended to increase, and T2 in the MFC significantly increased. Changes in cartilage thickness after 1 year showed a moderate correlation with the baseline T2 in the MFC as well as changes in T2 in the MTP. CONCLUSIONS: Decreasing cartilage thickness and matrix changes appeared in the MFC after 1 year of treatment with bDMARDs. Microstructural damage of the cartilage at baseline is a predictor for further cartilage damage in the knee joint, even if treatment with bDMARDs is effective.


Subject(s)
Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/diagnostic imaging , Cartilage, Articular/diagnostic imaging , Knee Joint/diagnostic imaging , Magnetic Resonance Imaging , Adolescent , Adult , Aged , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/pathology , Cartilage, Articular/drug effects , Cartilage, Articular/pathology , Female , Follow-Up Studies , Humans , Knee Joint/drug effects , Knee Joint/pathology , Magnetic Resonance Imaging/methods , Male , Middle Aged , Treatment Outcome , Young Adult
2.
Rev Sci Tech ; 37(2): 451-459, 2018 Aug.
Article in English | MEDLINE | ID: mdl-30747135

ABSTRACT

The effective control of dog rabies in Latin America is justifiably seen as a major success in the struggle to limit this devastating zoonosis. However, rabies remains a problem, due to the presence of the virus in bat populations throughout the region, including vampire bats. Vampire bats obtain nutrition exclusively through consuming blood by biting mammals and birds. This makes the species a highly efficient vector of the rabies virus, responsible for sporadic outbreaks of rabies in human populations and numerous cases of rabies in livestock. This, in turn, causes economic losses to the farming industry in countries throughout the region. For over four decades, efforts to control rabies have been directed at controlling the reservoir species and vaccinating cattle. However, this approach has not eliminated rabies in livestock. A major barrier to innovation in vampire rabies control is a lack of consistent surveillance to establish the extent of the problem. This precludes any calculation of its cost to the economy or the cost of potential solutions, such as vaccinating livestock. This paper outlines the problem of livestock rabies in Latin America and considers factors that influence the economic cost of potential solutions to this continuing challenge to human and livestock health.


Les résultats enregistrés en matière de lutte contre la rage canine en Amérique latine sont considérés avec raison comme un succès majeur sur la voie du contrôle de cette zoonose dévastatrice. Toutefois, la rage continue de poser un problème car le virus circule toujours parmi les populations de chauves-souris de la région, en particulier chez les chauves-souris hématophages. Ces dernières se nourrissent exclusivement de sang de mammifères ou d'oiseaux, en mordant leurs proies. Les espèces de chauves-souris hématophages sont donc des vecteurs extrêmement efficaces du virus de la rage et sont à l'origine de foyers sporadiques de rage dans les populations humaines ainsi que de nombreux cas de rage affectant le bétail. Cela occasionne des pertes économiques dans le secteur de l'élevage de tous les pays de la région. Depuis plus de quarante ans, les mesures appliquées pour lutter contre la rage ont essentiellement porté sur le contrôle des espèces réservoirs et sur la vaccination des bovins. Or, cette méthode n'a pas réussi à éliminer la rage chez le bétail. L'un des principaux obstacles à l'innovation en matière de lutte contre la rage chez les chauves-souris est qu'aucune surveillance n'est exercée de manière cohérente pour établir avec précision la portée du problème. Cela empêche d'évaluer le coût économique associé à la maladie ainsi que le coût des solutions envisageables, par exemple la vaccination du bétail. Les auteurs soulignent l'importance de la problématique de la rage chez les animaux d'élevage en Amérique latine et examinent les facteurs qui influent sur le coût économique des solutions à ce défi permanent pesant sur la santé humaine et celle du bétail.


El eficaz control de la rabia canina en América Latina se considera, con razón, un gran éxito en la lucha por contener esta devastadora zoonosis. La rabia, sin embargo, sigue planteando problemas por la presencia del virus en poblaciones de murciélagos de toda la región, en particular en murciélagos vampiro. Los vampiros se nutren exclusivamente de la sangre que sorben al morder a mamíferos y aves, lo que hace de ellos un vector sumamente eficaz del virus de la rabia, responsable de brotes esporádicos en poblaciones humanas y de numerosos casos de rabia bovina. Ello, a su vez, causa pérdidas económicas al sector pecuario de toda la región. Durante más de cuatro decenios, las medidas de lucha antirrábica han pasado esencialmente por el control de las especies que ejercen de reservorio y la vacunación de las reses. Esta lógica, sin embargo, no ha servido para acabar con la enfermedad en el ganado. Uno de los grandes obstáculos que impiden innovar en la lucha contra la rabia transmitida por los vampiros es la ausencia de una vigilancia sistemática que permita determinar el alcance del problema. Ello hace imposible calcular los costos que la enfermedad impone a la economía o el costo de eventuales soluciones, como la vacunación del ganado. Los autores exponen a grandes rasgos el problema de la rabia en el ganado latinoamericano y examinan los factores que influyen en el costo económico de las posibles soluciones a esta permanente amenaza para la salud de personas y animales.


Subject(s)
Cattle Diseases/pathology , Chiroptera , Disease Vectors , Rabies/veterinary , Animals , Caribbean Region/epidemiology , Cattle , Cattle Diseases/epidemiology , Cattle Diseases/virology , Humans , Latin America/epidemiology , Rabies/epidemiology , Rabies/pathology , Rabies virus , Zoonoses
4.
Bone Joint Res ; 5(7): 294-300, 2016 Jul.
Article in English | MEDLINE | ID: mdl-27421285

ABSTRACT

OBJECTIVES: The purpose of this study was to clarify the appearance of the reparative tissue on the articular surface and to analyse the properties of the reparative tissue after hemicallotasis osteotomy (HCO) using MRI T1ρ and T2 mapping. METHODS: Coronal T1ρ and T2 mapping and three-dimensional gradient-echo images were obtained from 20 subjects with medial knee osteoarthritis. We set the regions of interest (ROIs) on the full-thickness cartilage of the medial femoral condyle (MFC) and medial tibial plateau (MTP) of the knee and measured the cartilage thickness (mm) and T1ρ and T2 relaxation times (ms). Statistical analysis of time-dependent changes in the cartilage thickness and the T1ρ and T2 relaxation times was performed using one-way analysis of variance, and Scheffe's test was employed for post hoc multiple comparison. RESULTS: The cartilage-like repair tissue appeared on the cartilage surface of the medial compartment post-operatively, and the cartilage thickness showed a significant increase between the pre-operative and one-year post-operative time points (MFC; p = 0.003, MTP; p < 0.001). The T1ρ values of the cartilage-like repair tissue showed no difference over time, however, the T2 values showed a significant decrease between the pre-operative and one-year post-operative time points (MFC; p = 0.004, MTP; p = 0.040). CONCLUSION: This study clarified that the fibrocartilage-like repair tissue appeared on the articular surface of the medial compartment after HCO as evidenced by MRI T1ρ and T2 mapping.Cite this article: H. Nishioka, E. Nakamura, J. Hirose, N. Okamoto, S. Yamabe, H. Mizuta. MRI T1ρ and T2 mapping for the assessment of articular cartilage changes in patients with medial knee osteoarthritis after hemicallotasis osteotomy. Bone Joint Res 2016;5:294-300. DOI: 10.1302/2046-3758.57.BJR-2016-0057.R1.

7.
Haemophilia ; 21(5): 585-8, 2015 Sep.
Article in English | MEDLINE | ID: mdl-25708424

ABSTRACT

Combined thrombo-prophylaxis with mechanical and pharmacological methods is recommended in patients undergoing total knee or hip arthroplasty. As patients with 'untreated inherited bleeding disorders such as haemophilia' are at risk of bleeding, no prophylaxis has been prescribed for these patients. However, a retrospective study reported subclinical deep venous thrombosis (DVT) in 10% of patients with haemophilia undergoing major orthopaedic surgery. In this study, we aimed to evaluate the risk of DVT after total knee arthroplasty (TKA). We examined 38 TKA in 33 Japanese patients with haemophilia using ultrasonography. We did not detect DVT. The risk of DVT in patients with haemophilia after TKA may be lower than that in the general population. However, as patients with haemophilia progress in age, venous thromboembolism should be considered as a potential problem.


Subject(s)
Arthroplasty, Replacement, Knee/adverse effects , Asian People , Hemophilia A/complications , Venous Thrombosis/etiology , Adult , Aged , Factor VIII/metabolism , Humans , Middle Aged , Time Factors
8.
Osteoarthritis Cartilage ; 22(7): 1007-17, 2014 Jul.
Article in English | MEDLINE | ID: mdl-24795271

ABSTRACT

OBJECTIVE: When endoplasmic reticulum (ER) stress, i.e., the excessive accumulation of unfolded proteins in ER, endangers homeostasis, apoptosis is induced by C/EBP homologous protein (Chop). In osteoarthritis (OA) cartilage, Chop expression and apoptosis increase as degeneration progresses. We investigated the role of Chop in murine chondrocyte apoptosis and in the progression of cartilage degeneration. METHOD: We induced experimental OA in Chop-knockout (Chop(-/-)) mice by medial collateral ligament transection and meniscectomy and compared cartilage degeneration, apoptosis, and ER stress in Chop(-/-)- and wild-type (Chop(+/+)) mice. In our in vitro experiments we treated murine Chop(-/-) chondrocytes with the ER stress inducer tunicamycin (TM) and evaluated apoptosis, ER stress, and chondrocyte function. RESULTS: In vivo, the degree of ER stress was similar in Chop(-/-)- and Chop(+/+) mice. However, in Chop(-/-) mice apoptosis and cartilage degeneration were lower by 26.4% and 42.4% at 4 weeks, by 26.8% and 44.9% at 8 weeks, and by 26.9% and 32.3% at 12 weeks after surgery than Chop(+/+) mice, respectively. In vitro, the degree of ER stress induction by TM was similar in Chop(-/-)- and Chop(+/+) chondrocytes. On the other hand, apoptosis was 55.3% lower and the suppression of collagen type II and aggrecan mRNA was 21.0% and 23.3% less, and the increase of matrix metalloproteinase-13 mRNA was 20.0% less in Chop(-/-)- than Chop(+/+) chondrocytes. CONCLUSION: Our results indicate that Chop plays a direct role in chondrocyte apoptosis and that Chop-mediated apoptosis contributes to the progression of cartilage degeneration in mice.


Subject(s)
Apoptosis/physiology , Cartilage Diseases/pathology , Cartilage Diseases/physiopathology , Cartilage, Articular/pathology , Chondrocytes/pathology , Endoplasmic Reticulum Stress/physiology , Transcription Factor CHOP/physiology , Aggrecans/metabolism , Animals , Cartilage, Articular/physiopathology , Cells, Cultured , Chondrocytes/drug effects , Chondrocytes/metabolism , Collagen Type II/metabolism , Disease Models, Animal , Disease Progression , Endoplasmic Reticulum Stress/drug effects , Homeostasis/physiology , In Vitro Techniques , Matrix Metalloproteinase 13/metabolism , Mice , Mice, Inbred C57BL , Mice, Knockout , Transcription Factor CHOP/deficiency , Transcription Factor CHOP/genetics , Tunicamycin/pharmacology
9.
Bone Joint J ; 96-B(4): 467-72, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24692612

ABSTRACT

We report the long-term outcome of 33 patients (37 knees) who underwent proximal tibial open-wedge osteotomy with hemicallotasis (HCO) for medial osteoarthritis of the knee between 1995 and 2000. Among these, 29 patients with unilateral HCO were enrolled and 19 were available for review at a mean of 14.2 years (10 to 15.7) post-operatively. For these 19 patients, the mean Hospital for Special Surgery knee score was 60 (57 to 62) pre-operatively and 85 (82 to 87) at final follow-up (p < 0.001; paired t-test). The femorotibial angle and tibial inclination angle (IA) were measured at short-term follow-up, one to four years post-operatively, and showed no significant subsequent changes. The clinical scores and radiological measurements showed little change over time. One patient required conversion to total knee replacement during this time. These results suggest that the coronal angle achieved at operation is maintained at long-term follow up after HCO without alteration of the IA, providing a good long-term clinical outcome.


Subject(s)
Osteoarthritis, Knee/surgery , Osteogenesis, Distraction/methods , Osteotomy/methods , Tibia/surgery , Aged , Aged, 80 and over , External Fixators , Female , Follow-Up Studies , Humans , Knee Joint/diagnostic imaging , Knee Joint/pathology , Male , Middle Aged , Osteoarthritis, Knee/diagnostic imaging , Osteogenesis, Distraction/instrumentation , Prospective Studies , Radiography , Tibia/diagnostic imaging , Treatment Outcome
10.
Haemophilia ; 19(6): 951-5, 2013 Nov.
Article in English | MEDLINE | ID: mdl-23746133

ABSTRACT

Haemophilic arthropathy causes pain and a severely restricted range of motion, and results in a significant reduction in quality of life. When conservative treatments have failed, orthopaedic surgery is recommended for these patients with severe haemophilic arthropathy. However, surgery for haemophilia patients is challenging due to high complication rate such as infection, delayed wound healing and mortality. The aim of this study was to evaluate the incidence of early complications and identify preoperative risk factors of surgery for haemophilia patients. We report a series of haemophilia patients undergoing elective orthopaedic surgery between 2006 and 2012. During this period, 119 surgeries in 81 patients were prepared and 118 surgeries in 80 patients were actually performed. Four deep bacterial infections and four delayed wound healings occurred within 6 months postoperatively. One patient died preoperatively and four patients died postoperatively. Only the presence of inhibitor was a significant risk factor for infection. We found no risk factor related to delayed wound healing. Our data revealed alkaline phosphatase, albumin, platelet, alpha-fetoprotein, presence of ascites and child classification C as predictors of perioperative mortality following elective orthopaedic surgery. Our role is to identify potential patients who present with risk factors for complications and attempt to seek the best determination of treatment strategy for these people.


Subject(s)
Blood Coagulation Disorders, Inherited/complications , Joint Diseases/surgery , Adolescent , Adult , Aged , Bacterial Infections/etiology , Blood Coagulation Disorders, Inherited/mortality , Elective Surgical Procedures , Female , Humans , Japan , Joint Diseases/complications , Male , Middle Aged , Orthopedic Procedures , Postoperative Complications , Risk Factors , Splenomegaly/etiology , Wound Healing , Young Adult
11.
Arch Orthop Trauma Surg ; 128(12): 1447-52, 2008 Dec.
Article in English | MEDLINE | ID: mdl-18175132

ABSTRACT

INTRODUCTION: The Estimation of Physiologic Ability and Surgical Stress (E-PASS) scoring system is comprised of a preoperative risk score (PRS), a surgical stress score (SSS), and a comprehensive risk score (CRS) determined by both the PRS and SSS. E-PASS predicts the postoperative risk by quantifying the patient's reserve and surgical stress in general surgery. This study aims to evaluate the usefulness of this scoring system for the hospitalization outcomes in hip fracture. PATIENTS AND METHODS: A consecutive series of 419 elderly patients who underwent surgery with osteosynthesis or arthroplasty for hip fracture were prospectively assessed for the E-PASS scoring system, which was compared with their postoperative course. RESULTS: The postoperative morbidity and mortality rates in hospital increased linearly as the PRS and CRS increased, with significant correlation (rho = 0.2, P < 0.01) in both operations. The cost of hospital stay also related significantly to the SSS (r = 0.6, P < 0.0001) and CRS (r = 0.4, P < 0.0001). CONCLUSION: These results suggest that E-PASS may be useful for predicting postoperative risk and estimating medical expense for surgical cases with hip fracture.


Subject(s)
Fracture Fixation, Intramedullary/psychology , Fracture Healing/physiology , Health Status Indicators , Hip Fractures/surgery , Postoperative Complications/psychology , Adaptation, Physiological , Adaptation, Psychological , Age Factors , Aged , Aged, 80 and over , Cohort Studies , Female , Fracture Fixation, Intramedullary/methods , Fracture Fixation, Intramedullary/mortality , Geriatric Assessment , Hip Fractures/mortality , Hip Fractures/psychology , Hospital Costs , Humans , Length of Stay/economics , Male , Postoperative Complications/etiology , Postoperative Complications/mortality , Predictive Value of Tests , Probability , Prospective Studies , Recovery of Function , Risk Assessment , Statistics, Nonparametric , Stress, Physiological , Survival Analysis
12.
J Bone Joint Surg Br ; 89(5): 693-700, 2007 May.
Article in English | MEDLINE | ID: mdl-17540759

ABSTRACT

We have investigated in vitro the release kinetics and bioactivity of fibroblast growth factor-2 (FGF-2) released from a carrier of fibrin sealant. In order to evaluate the effects of the FGF-2 delivery mechanism on the repair of articular cartilage, full-thickness cylindrical defects, 5 mm in diameter and 4 mm in depth, which were too large to undergo spontaneous repair, were created in the femoral trochlea of rabbit knees. These defects were then filled with the sealant. Approximately 50% of the FGF-2 was released from the sealant within 24 hours while its original bioactivity was maintained. The implantation of the fibrin sealant incorporating FGF-2 successfully induced healing of the surface with hyaline cartilage and concomitant repair of the subchondral bone at eight weeks after the creation of the defect. Our findings suggest that this delivery method for FGF-2 may be useful for promoting regenerative repair of full-thickness defects of articular cartilage in humans.


Subject(s)
Cartilage, Articular/injuries , Drug Delivery Systems/methods , Fibrin Tissue Adhesive/administration & dosage , Fibroblast Growth Factor 2/administration & dosage , Wound Healing/drug effects , Animals , Cartilage, Articular/drug effects , Cartilage, Articular/pathology , Cartilage, Articular/physiology , Cell Proliferation/drug effects , Cells, Cultured , Collagen Type II/metabolism , Drug Evaluation, Preclinical , Fibrin Tissue Adhesive/therapeutic use , Fibroblast Growth Factor 2/pharmacokinetics , Fibroblast Growth Factor 2/therapeutic use , Fibroblasts/cytology , Fibroblasts/drug effects , Male , Mice , Mice, Inbred BALB C , Rabbits
13.
Bioresour Technol ; 84(3): 287-90, 2002 Sep.
Article in English | MEDLINE | ID: mdl-12118708

ABSTRACT

Pectin had a flocculating activity and its flocculating activities in various suspensions were investigated. Flocculating activity of pectin in a kaolin suspension was markedly stimulated by the addition of Al3+ and Fe3+ to the suspension. Optimum temperature for flocculating activity of pectin in the kaolin suspension was around 30 degrees C and high flocculating activity was obtained when 30 mg/l of pectin and 0.2 mM Fe3+ were added to the suspension. Other inorganic suspensions of activated carbon and acid clay were flocculated by pectin in the presence of Al3+ or Fe3+. Flocculation of organic suspensions such as cellulose and yeast by pectin occurred when 0.1-0.2 mM Fe3+ was present in the suspensions.


Subject(s)
Pectins/chemistry , Aluminum/analysis , Bioreactors , Biotechnology/methods , Cations , Dose-Response Relationship, Drug , Flocculation , Iron/analysis , Temperature
14.
Acta Orthop Scand ; 72(4): 404-10, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11580130

ABSTRACT

Oxygen free radicals (OFR) and neutrophils are potent sources of reperfusion injury. We compared the effect of EPC-K1, a new OFR scavenger, and neutrophil depletion on the reperfusion injury in skeletal muscle, using an ischemic revascularized hindlimb model in rats. Warm ischemia, produced by vascular pedicle clamping, was sustained for 4 h. After 24 h of reperfusion, muscle function and damage were evaluated in 4 groups: a sham operation group, a control study group, a group treated by EPC-K1 (EPC group), and a group that received nitrogen mustard to induce neutropenia (NM group). Both the EPC and NM groups had limited muscle damage compared to the control group. The EPC group preserved muscle function significantly better than the control group and the mean isometric tetanic tension in the EPC group appeared to be higher than that in the NM group. Furthermore, levels of lipid peroxides in muscle and serum, and muscle edema in the EPC group, were significantly lower than in the NM group. Histological examinations supported these results. These findings suggest that limiting OFR generation by EPC-K1 in the early phase of reoxygenation is more potent than depletion of neutrophils in reducing reperfusion injury.


Subject(s)
Ascorbic Acid/analogs & derivatives , Ascorbic Acid/therapeutic use , Disease Models, Animal , Free Radical Scavengers/therapeutic use , Muscle, Skeletal/injuries , Neutropenia/etiology , Reperfusion Injury/drug therapy , Reperfusion Injury/immunology , Vitamin E/analogs & derivatives , Vitamin E/therapeutic use , Animals , Ascorbic Acid/chemistry , Ascorbic Acid/immunology , Drug Evaluation, Preclinical , Free Radical Scavengers/chemistry , Free Radical Scavengers/immunology , Isometric Contraction/drug effects , Leukocyte Count , Lipid Peroxidation/drug effects , Male , Neutropenia/blood , Neutrophils/drug effects , Random Allocation , Rats , Rats, Inbred Lew , Reperfusion Injury/complications , Reperfusion Injury/metabolism , Vitamin E/chemistry , Vitamin E/immunology
15.
Biochemistry ; 40(39): 11860-5, 2001 Oct 02.
Article in English | MEDLINE | ID: mdl-11570886

ABSTRACT

Dipeptidyl peptidase III (DPP III) (EC 3.4.14.4), which has a HELLGH-E (residues 450-455, 508) motif as the zinc binding site, is classified as a zinc metallopeptidase. The zinc dissociation constants of the wild type, Leu(453)-deleted, and E508D mutant of DPP III at pH 7.4 were 4.5 (+/-0.7) x 10(-13), 5.8 (+/-0.7) x 10(-12), and 3.2 (+/-0.9) x 10(-10) M, respectively. The recoveries of the enzyme activities by the addition of various metal ions to apo-DPP III were also measured, and Co(2+), Ni(2+), and Cu(2+) ions completely recovered the enzyme activities as did Zn(2+). The dissociation constants of Co(2+), Ni(2+), and Cu(2+) ions for apo-DPP III at pH 7.4 were 8.2 (+/-0.9) x 10(-13), 2.7 (+/-0.3) x 10(-12), and 1.1 (+/-0.1) x 10(-14) M, respectively. The shape of the absorption spectrum of Co(2+)-DPP III was very similar to that of Co(2+)-carboxypeptidase A or Co(2+)-thermolysin, in which the Co(2+) is bound to two histidyl nitrogens, a water molecule, and a glutamate residue. The absorption spectrum of Cu(2+)-DPP III is also very similar to that of Cu(2+)-thermolysin. The EPR spectrum and the EPR parameters of Cu(2+)-DPP III were very similar to those of Cu(2+)-thermolysin but slightly different from those of Cu(2+)-carboxypeptidase A. The five lines of the superfine structure in the perpendicular region of the EPR spectrum in Cu(2+)-DPP III suggest that nitrogen atoms should coordinate to the cupric ion in Cu(2+)-DPP III. All of these data suggest that the donor set and the coordination geometry of the metal ions in DPP III, which has the HExxxH motif as the metal binding site, are very similar to those of the metal ions in thermolysin, which has the HExxH motif.


Subject(s)
Dipeptidyl-Peptidases and Tripeptidyl-Peptidases/metabolism , Liver/enzymology , Zinc/metabolism , Animals , Electron Spin Resonance Spectroscopy , Kinetics , Rats , Recombinant Proteins/metabolism
16.
Biochem J ; 359(Pt 1): 99-108, 2001 Oct 01.
Article in English | MEDLINE | ID: mdl-11563973

ABSTRACT

We identified a novel Drosophila protein of approximately 400 kDa, hemolectin (d-Hml), secreted from haemocyte-derived Kc167 cells. Its 11.7 kbp cDNA contains an open reading frame of 3843 amino acid residues, with conserved domains in von Willebrand factor (VWF), coagulation factor V/VIII and complement factors. The d-hml gene is located on the third chromosome (position 70C1-5) and consists of 26 exons. The major part of d-Hml consists of well-known motifs with the organization: CP1-EG1-CP2-EG2-CP3-VD1-VD2-VD'-VD3-VC1-VD"-VD"'-FC1-FC2-VC2-LA1-VD4-VD5-VC3-VB1-VB2-VC4-VC5-CK1 (CP, complement-control protein domain; EG, epidermal-growth-factor-like domain; VB, VC, VD, VWF type B-, C- and D-like domains; VD', VD", VD"', truncated C-terminal VDs; FC, coagulation factor V/VIII type C domain; LA, low-density-lipoprotein-receptor class A domain; CK, cysteine knot domain). The organization of VD1-VD2-VD'-VD3, essential for VWF to be processed by furin, to bind to coagulation factor VIII and to form interchain disulphide linkages, is conserved. The 400 kDa form of d-Hml was sensitive to acidic cleavage near the boundary between VD2 and VD', where the cleavage site of pro-VWF is located. Agarose-gel electrophoresis of metabolically radiolabelled d-Hml suggested that it is secreted from Kc167 cells mainly as dimers. Resembling VWF, 7.9% (305 residues) of cysteine residues on the d-Hml sequence had well-conserved positions in each motif. Coinciding with the development of phagocytic haemocytes, d-hml transcript was detected in late embryos and larvae. Its low-level expression in adult flies was induced by injury at any position on the body.


Subject(s)
Drosophila Proteins/genetics , Drosophila melanogaster/metabolism , Hemocytes/metabolism , Lectins/genetics , Amino Acid Sequence , Animals , Base Sequence , Blotting, Northern , Cells, Cultured , Chromosomes/genetics , Cloning, Molecular , DNA Primers/chemistry , DNA, Complementary/genetics , DNA, Complementary/metabolism , Disulfides/metabolism , Drosophila Proteins/chemistry , Drosophila Proteins/metabolism , Electrophoresis, Polyacrylamide Gel , Gene Expression Regulation, Developmental , Humans , Lectins/chemistry , Lectins/metabolism , Molecular Sequence Data , Peptide Fragments/immunology , Polymerase Chain Reaction , Protein Binding , Protein Structure, Tertiary , Rabbits , Recombinant Proteins/metabolism , Sequence Homology, Amino Acid , von Willebrand Factor/chemistry
17.
Biosci Biotechnol Biochem ; 65(6): 1438-40, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11471751

ABSTRACT

The major food allergen, ovomucoid (molecular weight of 28 kDa) could be detected in 12 of 37 human breast milk samples by using three types of enzyme-linked immunosorbent assay. By gel-filtration, ovomucoid in breast milk was only eluted in the fractions corresponding to a molecular weight of about 450 kDa, suggesting its occurrence as an immune complex with IgA. In fact, almost the same elution profile as that for ovomucoid was obtained for its immune complex with IgA by gel-filtration.


Subject(s)
Allergens/analysis , Antigen-Antibody Complex/chemistry , Food Hypersensitivity/immunology , Milk, Human/chemistry , Ovomucin/analysis , Adult , Animals , Chromatography, Gel , Eggs/analysis , Female , Humans , Mice , Mice, Inbred BALB C , Rats
18.
J Surg Res ; 99(1): 1-6, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11421597

ABSTRACT

BACKGROUND: The ultimate aim of replantations and transplantations of skeletal muscle is to improve impaired function. The purpose of this study was to examine the contribution of varying durations of ischemia to postischemic blood flow in the skeletal muscle and the contribution of modulation of postischemic blood flow to skeletal muscle function and viability, using an ischemic revascularized hind limb model in rats. MATERIALS AND METHODS: Warm ischemia produced by vascular pedicle clamping was sustained for 90 min, 3 h, or 6 h. Postischemic blood flow was measured by a Doppler flowmeter or microsphere technique. In another series of experiments of 3-h ischemia, either saline or N(G)-methyl-l-arginine acetate (l-NMMA) was infused for the first 2 h of reperfusion. Postischemic blood flow was also measured. Muscle contractile function and viability were determined after 24 h of reperfusion. RESULTS: Postischemic blood flow was significantly increased during the first 10 min of reperfusion in the 90-minute ischemic group and during the first 2 h in the 3-h ischemic group compared with contralateral control blood flow. No significant increase in postischemic blood flow was noted in the 6-h ischemic group. Postischemic blood flow was significantly decreased by the l-NMMA infusion. Contractile function and viability of the tibialis anterior muscle and contractile function of the gastrocnemius muscle in the l-NMMA group were significantly increased. CONCLUSION: Reperfusion blood flow increased time dependently until 3 h of warm ischemia. Hyperemia deteriorated skeletal muscle contractile function, although it was well preserved by l-NMMA infusion to restrict the postischemic hyperemia.


Subject(s)
Ischemia/physiopathology , Isometric Contraction , Muscle, Skeletal/blood supply , Muscle, Skeletal/physiopathology , Reperfusion Injury/physiopathology , Animals , Enzyme Inhibitors/pharmacology , Hindlimb , Hyperemia/physiopathology , Isometric Contraction/drug effects , Laser-Doppler Flowmetry , Male , Microspheres , Muscle, Skeletal/drug effects , Rats , Rats, Inbred Lew , Regional Blood Flow , Tissue Survival/drug effects , omega-N-Methylarginine/pharmacology
19.
Biosci Biotechnol Biochem ; 65(3): 658-61, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11330684

ABSTRACT

A mannose isomerase from Agrobacterium radiobacter M-1 (formerly Pseudomonas sp. MI) was purified to electrophoretic homogeneity and characterized. A cell-free extract was separated by ammonium sulfate fractionation, Butyl-Toyopearl 650M, DEAE-Sepharose and hydroxylapatite column chromatography. Its molecular mass was estimated to be 44 kDa by SDS-PAGE and 90 kDa by gel filtration, in which the enzyme is most likely a dimer composed of two identical subunits. The purified enzyme had an optimum pH at 8.0, an optimum temperature at 60 degrees C, a pI of 5.2 and a Km of 20 mM, and specifically converted D-mannose and D-lyxose to ketose. The N-terminal amino acid sequence was identified.


Subject(s)
Aldose-Ketose Isomerases/analysis , Rhizobium/enzymology , Aldose-Ketose Isomerases/antagonists & inhibitors , Cations, Divalent , Enzyme Inhibitors/pharmacology , Enzyme Stability , Hydrogen-Ion Concentration , Metals , Molecular Weight , Substrate Specificity , Temperature
20.
J Biol Chem ; 276(7): 5228-34, 2001 Feb 16.
Article in English | MEDLINE | ID: mdl-11083865

ABSTRACT

We previously reported that versican, a large chondroitin sulfate proteoglycan, isolated from a renal adenocarcinoma cell line, ACHN, binds L-selectin. Here we report that versican also binds certain chemokines and regulates chemokine function. This binding was strongly inhibited by the chondroitinase digestion of versican or by the addition of soluble chondroitin sulfate (CS) B, CS E, or heparan sulfate. Furthermore, these glycosaminoglycans (GAGs) could bind directly to the chemokines that bind versican. Thus, versican appears to interact with chemokines via its GAGs. We next examined if versican or GAGs affect secondary lymphoid tissue chemokine (SLC)-induced integrin activation and Ca(2+) mobilization in lymphoid cells expressing a receptor for SLC, CC chemokine receptor 7. Interestingly, whereas heparan sulfate supported both alpha(4)beta(7) integrin-dependent binding to mucosal addressin cell adhesion molecule-1 (MAdCAM-1)-IgG and Ca(2+) mobilization induced by SLC, versican or CS B inhibited these cellular responses, and the extent of inhibition was dependent on the dose of versican or CS B added. These findings suggest that different proteoglycans have different functions in the regulation of chemokine activities and that versican may negatively regulate the function of SLC via its GAG chains.


Subject(s)
Chemokines/metabolism , Chondroitin Sulfate Proteoglycans/physiology , Animals , Calcium/metabolism , Cell Adhesion , Chemokine CCL21 , Chemokines/pharmacology , Chemokines, CC/metabolism , Chemokines, CC/pharmacology , Chondroitin Sulfate Proteoglycans/chemistry , Chondroitin Sulfate Proteoglycans/metabolism , Dermatan Sulfate/metabolism , Dermatan Sulfate/pharmacology , Glycosaminoglycans/metabolism , Glycosaminoglycans/pharmacology , Humans , Integrins/physiology , Jurkat Cells , Lectins, C-Type , Mice , T-Lymphocytes/metabolism , Versicans
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