Separable regulatory elements from pro-alpha 2 (XI) collagen gene drive distinct patterns of cartilage-specific expression in transgenic mice.
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Animals↗
Biomedical subjects
T Ochi
Publications and source records attributed to T Ochi.
Transient introduction of a foreign gene into healing rat patellar ligament.
We investigated the in vivo introduction of a reporter gene into healing rat patellar ligaments using the hemagglutinating virus of Japan (HVJ)-liposome-mediated gene transfer method. The mid-portion of the medial half of the patellar ligament was cut transversely with a scalpel in 14-wk-old male Wistar rats. A HVJ-liposome suspension containing beta-galactosidase (beta-gal) cDNA was injected directly into the injured site and pooled in the fascial pocket covering the injured site 3 d postoperatively. Thereafter, beta-gal-labeled cells were observed in the wound site accounting for 3% of the wound cells on the first day, 2% on the third, 7% on the seventh, 6% on the 14th, 2% on the 28th, and 0.2% on the 56th day after injection. The beta-gal-labeled cells were initially localized in and adjacent to the wound site, but they were observed spreading into the ligament substance away from the wound on the seventh day after injection. On day 28, beta-gal-labeled cells were observed throughout the length of the ligament substance. With double-labeling for marker antigens for monocyte/macrophage (ED-1) and for collagen I aminopropeptide (pN collagen I), it was revealed that fibroblastic (pN collagen I-positive) cells accounted for 63% and monocyte/macrophage lineage cells for 32% of the beta-gal-labeled cells in the day 7 wound. On day 28, they formed 58 and 35% of the beta-gal-labeled cells in the wound, respectively. Thus, we succeeded in introducing the beta-gal gene into healing rat patellar ligament. Moreover, labeling of the transfected cells made it possible to identify a biological event, namely that the cells in and around the wound site infiltrate into the uninjured ligament substance and come to populate the whole length of the ligament substance as repair progresses. These results suggest that ligament healing may involve not only the repair of the wound site itself but also extensive cellular infiltration of ligament substance adjacent to the wound.
Animals↗
Elevated levels of the soluble form of bone marrow stromal cell antigen 1 in the sera of patients with severe rheumatoid arthritis.
OBJECTIVE: Bone marrow stromal cell antigen 1 (BST-1) is a novel glycosyl phosphatidylinositol-anchored ectoenzyme, which is overexpressed on bone marrow stromal and synovial cell lines derived from patients with rheumatoid arthritis (RA). To investigate the pathophysiologic roles of BST-1 in RA, we established an enzyme-linked immunosorbent assay (ELISA) system to detect the soluble form of BST-1 (sBST-1) and examined levels of sBST-1 in the sera of RA patients. METHODS: Concentrations of sBST-1 in sera from healthy donors and from patients with RA, osteoarthritis, Sjögren's syndrome, and systemic lupus erythematosus were measured with the ELISA. RESULTS: In 7% of the RA patient samples (10 of 143), concentrations of serum sBST-1 were higher (approximately 30-50-fold) than in non-RA samples. Serum sBST-1 concentrations showed no correlation with age, C-reactive protein level, or rheumatoid factor level. All RA patients with high concentrations of serum sBST-1 had severe disease with involvement of several large joints. CONCLUSION: We believe the measurement of serum sBST-1 may have prognostic value, but further analysis is necessary to clarify the clinical significance of elevated sBST-1 in RA.
ADP-ribosyl Cyclase↗
Accelerated generation of CD14+ monocyte-lineage cells from the bone marrow of rheumatoid arthritis patients.
OBJECTIVE: To examine the capacity of bone marrow progenitor cells to generate CD14+ cells, in order to assess the role of bone marrow in the pathogenesis of rheumatoid arthritis (RA). METHODS: CD14- cells purified from bone marrow specimens of 11 patients with active RA and 8 control patients (osteoarthritis or trauma) were cultured in the presence or absence of granulocyte-macrophage colony-stimulating factor (GM-CSF; 100 pg/ml). After incubation for various lengths of time, the cells were analyzed by flow cytometry for expression of CD14 and HLA-DR. RESULTS: The spontaneous generation of CD14+ cells from bone marrow CD14- progenitor cells was accelerated in RA patients compared with control patients. Moreover, the expression of HLA-DR on the bone marrow-derived CD14+ cells was also accelerated in RA patients compared with controls. GM-CSF significantly enhanced the generation of CD14+ cells, as well as the expression of HLA-DR, on CD14+ cells of control patients, but not those of RA patients. GM-CSF levels in the culture supernatants of bone marrow CD14- cells were not significantly different between RA patients and control patients (undetectable in most cases). CONCLUSION: These observations strongly support the hypothesis that the accelerated generation of CD14+ cells from bone marrow progenitor cells and the accelerated maturation of such CD14+ cells into HLA-DR+ cells play an important role in the pathogenesis of RA. Moreover, the data suggest a functional alteration of RA bone marrow CD14- cells in their responsiveness to GM-CSF.
Adult↗
A recurrent 1992delCT mutation of the type X collagen gene in a Japanese patient with Schmid metaphyseal chondrodysplasia.
We report here a recurrent frameshift mutation within the carboxyl-terminal noncollagenous domain coding region of the type X collagen gene (COL10A1) in a Japanese patient with Schmid metaphyseal chondrodysplasia. The mutation involves deletion of a CT dinucleotide from position 1992 (1992delCT), and produces a frameshift which creates a premature termination codon close to the site of the deletion. The predicted length of the mutant polypeptide is 664 amino acids, which is shorter than the wild type polypeptide (680 amino acids). A 1992delCT mutation of COL10A1 has been previously reported in one family. The independent occurrence of de novo mutation of this specific dinucleotide repeat suggests that this region is a possible mutational hot spot on COL10A1.
Asian People↗
Dimethylarsinic acid causes apoptosis in HL-60 cells via interaction with glutathione.
Inducibility of apoptosis in cultured human HL-60 cells by arsenic compounds, such as arsenite, arsenate, methylarsonic acid (MAA), and dimethylarsinic acid (DMAA), was investigated, together with the role of glutathione (GSH) in the induction. Among the arsenic compounds DMAA was the most potent in terms of the ability to cause the morphological changes (formation of nuclear fragmentation and apoptotic bodies) characteristic of apoptosis. Furthermore, fragmentation of internucleosomal DNA was also induced by DMAA. Depletion of cell GSH by L-buthionine-SR-sulfoximine, a selective inhibitor of gamma-glutamylcysteine synthetase, enhanced the cytotoxicity of arsenite, arsenate, and MAA, while such depletion suppressed the cytotoxicity of DMAA. The depletion of GSH also suppressed the morphological changes and the fragmentation of internucleosomal DNA caused by DMAA, both of which are characteristic features of apoptosis. The results suggest that the death of cells caused by DMAA is due to apoptosis and that GSH is involved in the induction of apoptosis by this arsenic compound.
Apoptosis↗
Acetabular labral tears: contrast-enhanced MR imaging under continuous leg traction.
OBJECTIVE: The objective of this study was to evaluate the effects of continuous leg traction on contrast-enhanced MR imaging of the hip joint and to determine whether MR imaging under these conditions is useful for demonstrating acetabular labral tears. DESIGN AND PATIENTS: Nineteen hips underwent MR imaging with a T1-weighted spin-echo sequence, followed by MR imaging under continuous leg traction after intravenous injection of gadolinium-DTPA. Joint fluid enhancement and labral contour detection were evaluated. Eleven hips had labral tears shown by conventional arthrography, arthroscopy and macroscopic surgical findings. Assessment of labral tears by MR imaging was correlated with the diagnosis based on these standard techniques. RESULTS AND CONCLUSIONS: Joint fluid enhancement was obtained in all hips at 30 min after injection. Superior and inferior labral surfaces were completely delineated in 1 hip on the unenhanced MR images, and in 7 and 13 hips, respectively, on the enhanced images under traction. The enhanced images under traction depicted 9 of the 11 labral tears. Comparison between the unenhanced image and the enhanced image under traction avoided mistaking undercutting of the labrum for a tear in 4 hips. Contrast-enhanced MR imaging under traction was valuable for detecting labral tears non-invasively and without radiation. Follow-up examinations using this method in patients with acetabular dysplasia can help to clarify the natural course of labral disorders and enable better treatment planning.
Acetabulum↗
MR arthrography of elbow: evaluation of the ulnar collateral ligament of elbow.
OBJECTIVE: The purpose of this study was to evaluate ulnar collateral ligament (UCL) injury of the elbow in throwing athletes by MRI and MR arthrography. DESIGN: Ten elbows of throwing athletes were examined on both plain MRI and MR saline arthrography and the injuries subsequently surgically proven. Spin-echo (SE) T1-weighted and fast SE T2-weighted coronal images were obtained. RESULTS: The UCL was unclear in all ten cases on T1-weighted MRI. In five cases an avulsion fracture was also found on T1-weighted MRI. On T2-weighted MRI, abnormal high-intensity areas were identified in or around the UCL. On T2-weighted MR arthrography images, extracapsular high-intensity areas, which represent extracapsular leakage, were found in four of five cases with avulsion fracture. At surgery, all these four cases showed avulsion fractures with instability; the other case had a fracture but it was stable and adherent to the humerus. On T2-weighted MR arthrography images, an extracapsular high-intensity area was found in one of the five cases without avulsion fracture. At surgery this patient had a complete tear of the UCL itself. CONCLUSION: MR arthrography provided additional information for evaluating the degree of UCL injury.
Adolescent↗
Progressive degeneration of articular cartilage and intervertebral discs. An experimental study in transgenic mice bearing a type IX collagen mutation.
Transgenic mice expressing mutant alpha 1(IX) collagen were produced and found to develop progressive joint degeneration with age, as well as accelerated intervertebral disc degeneration. Radiological and histological studies showed that cervical and lumbar disc degeneration was more advanced in the transgenic mice than in control litter-mates. The changes included shrinkage or disappearance of the nucleus pulposus, and fissures in the annulus fibrosus which sometimes lead to herniation of disc material and slight osteophyte formation. These findings suggest that mutations of the type IX collagen may cause certain forms of degenerative disease in the spine as well as in joints.
Animals↗
Inducible osteonecrosis in a rabbit serum sickness model: deposition of immune complexes in bone marrow.
We established inducible osteonecrosis in a rabbit serum sickness model. Osteonecrosis with marrow necrosis could be induced by the intravenous injection of horse serum in two doses separated in time by a period of three weeks. In this model, osteonecrosis could be successfully produced in rabbit femoral metaphysis. The incidence of marrow necrosis was 45% (9 of 20 rabbits) and trabecular necrosis occurred in 6 of 20 rabbits (30%) at 7 days after the second injection of the horse serum. In bone marrow of the femoral metaphysis, extravasation of erythrocytes and the formation of micro-thrombi in arterioles were often observed in an early stage of the present model and both findings correlate well each other (p = 0.0001). Immune complexes could be demonstrated using immunohistochemistry in bone marrow of the femoral metaphysis as well as in glomeruli of the kidney. Extravasation of erythrocytes in bone marrow of the femoral metaphysis was observed in 8 of 12 (67%) cases with immune complex deposition in the sinusoidal space of the femoral metaphysis and in 12 of 21 (57%) cases with immune complex deposition in glomeruli of the kidney. Immune complex deposition both in the sinusoidal space of femoral bone marrow (p = 0.0385) and in glomeruli of the kidney (p = 0.0209) closely related to extravasation of erythrocytes and microthrombi in arterioles in the early stage of this model. Early microcirculatory injury (extravasation of erythrocytes and microthrombi in arterioles) adjacent to osteonecrosis could be induced by immune complex deposition in femoral bone marrow and might be predictable characteristics for the inducible osteonecrosis in the present serum sickness model. The important findings in this study were that early microcirculatory injury was closely related to the deposition of immune complexes in femoral bone marrow, and that early microcirculatory injury associated with immune complex deposition was located close to osteonecrotic regions.
Animals↗
Results of isolated meniscal repair evaluated by second-look arthroscopy.
Thirty-six isolated torn menisci in 35 patients (average age, 24 years) which had been repaired arthroscopically using an inside-out technique were evaluated by second-look arthroscopy. The time from meniscal repair to second-look arthroscopy ranged from 2 to 10 months with a mean of 5 months. The indications for meniscal repair were a longitudinal or oblique tear located at the outer half of the meniscus. Twenty (56%) were graded as excellent, 10 (28%) as good, and 6 (16%) were graded as poor. Neither age nor length of time between injury and repair affected meniscal healing. The medial meniscal repairs showed better results than the lateral repairs (rate of excellent results: medial, 82%; lateral, 44%; P < .01, chi-squared test). The rate of excellent results for those with normal meniscal bodies at the time of repair was 79%, which was significantly higher than that seen in the cases with deformed and/or superficial damage to the meniscal body (36%; P < .05, chi-squared test).
Adolescent↗
Separable cis-regulatory elements that contribute to tissue- and site-specific alpha 2(XI) collagen gene expression in the embryonic mouse cartilage.
Type XI collagen is a structural component of the cartilage extracellular matrix and plays an important role in skeletal morphogenesis. As a step toward defining the molecular mechanisms responsible for the regulation of type XI collagen expression, we characterized the promoter region of the mouse alpha 2(XI) collagen gene (Coll1a2). We also generated transgenic mice harboring various fragments of the promoter and the first intron of Coll1a2 linked to the Escherichia coli beta-galactosidase gene to identify the cis-acting elements responsible for tissue- and site-specific expression during development. Cloning and sequence analysis of the 5' flanking region of Coll1a2 showed that the putative 3' end of the retinoid X receptor beta gene was located 742 bp upstream of the Coll1a2 start site. This suggested that the promoter region of Coll1a2 was localized within this 742-bp sequence, which contained multiple consensus regulatory elements. Examination of the transgenic mice revealed that the longest DNA construct (containing the entire promoter and first intron sequences) directed lacZ expression in the notochord as well as in the primordial cartilage throughout the body, with the pattern of expression mimicking that of endogenous Coll1a2 transcripts. On the other hand, deletion of the upstream approximately 290 bp resulted in the elimination of lacZ expression in the primordial cartilage of the carpals, tarsals, and vertebral bodies, whereas lacZ expression in the notochord and in the other primordial cartilage elsewhere was not affected. Deletion of the first intron sequence also resulted in the loss of lacZ expression in the primordial cartilage of the carpals, tarsals, and vertebral bodies, as well as in the notochord. These results demonstrate that the upstream 742-bp and first intron segments of the mouse Coll1a2 gene contain the necessary information to confer high level tissue-specific expression in mouse embryos. In addition, our observations suggest the presence of site-specific cis-acting elements that control Coll11a2 gene expression in different cartilaginous components of the skeleton.
Animals↗
Shoulder destruction in rheumatoid arthritis. Classification and prognostic signs in 83 patients followed 5-23 years.
We studied the natural course and the possibility of making prognoses about shoulder joint destructions in 83 patients with rheumatoid arthritis (RA) (166 shoulder joints). For this purpose, we used radiographic patterns and 2 indices (upward migration and medial displacement). The patterns of joint destruction were classified into 5 groups: 1) non-progressive type (n 74) with normal radiographs, only osteopenia or small erosions even after 15-20 years of RA; 2) erosive type (n 22) showing marginal erosions but no collapse; 3) collapse type (n 34) showing subchondral cysts, followed by collapse; 4) arthrosis-like type (n 12) showing arthrotic features; 5) mutilating type (n 14) showing mutilating bone destructions. From the radiographic findings and the 2 indices determined at 5-10 years, we could predict the prognosis of shoulder joint destruction after 15-20 years of RA. Our findings may be of value for selecting treatment, including surgery, for the rheumatoid shoulder.
Adult↗
MRI of early osteonecrosis of the femoral head after transcervical fracture.
We have carried out a prospective study of 17 patients (14 women, 3 men) of mean age 48 years (21 to 76) with transcervical fractures of the femur using MRI to detect early evidence of avascular necrosis of the head. Two fractures were Garden stage I, 12 stage II, and three stage III. We performed internal fixation under radiological control at a mean of five days (2 to 15) after injury using a titanium cannulated cancellous screw or a titanium compression hip screw. MRI was performed at one, six and 12 months and then yearly after operation. T1- and T2-weighted images were obtained by a spin-echo technique. The duration of follow-up of patients who did not subsequently require replacement of the head of the femur was from 2 to 5 years (mean 3.2). One month after operation eight of the 17 hips showed a band of low signal intensity on T1-weighted images and high signal intensity on T2-weighted images indicating lesions in the femoral head away from the fracture line. These were of three types: type I was a small infarct at the superolateral region of the femoral head and was seen in three hips; type II was a shallow lesion from the superolateral region to the fovea of the femoral head (three hips); and type III was a large lesion occupying most of the femoral head (two hips). No further changes were seen in the MRI after six months from operation. Collapse of the femoral head did not occur in the three hips with type-I lesions, but two of the three type-II hips and both type-III hips subsequently collapsed. At the final follow-up the three hips with a type-I lesion and one with a type-II were still asymptomatic but radiography showed sclerosis in the femoral head corresponding to the MRI lesions. The nine hips which showed no changes on MRI at one month had no abnormal findings on physical examination, radiography or MRI at final follow-up.
Adult↗
Effect of alacepril on 24-hour blood pressure in elderly hypertensive patients.
The effect of administration of an angiotensin-converting enzyme inhibitor, alacepril, on 24-hour blood pressure in the elderly hypertensive patients was investigated. Thirteen elderly hypertensive patients (mean age 71 +/- 5; 6 male and 7 female) participated in the present study. After 2 weeks of control period alacepril was administered 25-50 mg/day for 8 weeks. Ambulatory blood pressure monitoring with cuff-oscillometric method was performed at the end of both control and treatment periods. Alacepril administration for 8 weeks significantly decreased 24-hour blood pressure while it had little effect on heart rate. Daytime blood pressure was significantly decreased from 154 +/- 10/91 +/- 5 mmHg to 145 +/- 8/85 +/- 5 mmHg, while the change in nocturnal blood pressure was not significant: from 137 +/- 17/79 +/- 7 mmHg to 130 +/- 15/75 +/- 9 mmHg. Hyperbaric area of systolic blood pressure was also significantly decreased (from 295 +/- 185 mmHg x hour/day to 172 +/- 111 mmHg x hour/day), indicating that pressure load to the heart was effectively reduced. Administration of alacepril did not cause tachycardia in response to the decrease in blood pressure. Acrophase of both blood pressure and heart rate was changed to 11:00 a.m. These findings indicate that blockade of the renin-angiotensin system in the elderly hypertensive patients decreased blood pressure effectively without causing tachycardia or deterioration of diurnal variations of blood pressure. These hemodynamic changes produced by alacepril administration are favorable for the treatment of the elderly patients with cardiovascular disease.
Aged↗
[Pathology and management of spinal lesions in rheumatoid arthritis].
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Anti-Inflammatory Agents↗
Bone marrow stromal cells contribute to synovial cell proliferation in rats with collagen induced arthritis.
OBJECTIVE: To observe the migration of bone marrow stromal cells into the joint cavity and the contribution of such cells to synovial proliferation in rats with collagen induced arthritis (CIA). METHODS: After bone marrow stromal cells preliminarily labeled with fluorescent dye or 3H thymidine accumulated in the bone marrow of splenectomized rats by intraperitoneal injection, the migration of labeled stromal cells in rats with CIA was analyzed by liquid scintillation counting, autoradiography, and fluorography. RESULTS: In splenectomized control rats, labeled bone marrow stromal cells were mostly found in the bone marrow and not in the synovium. Over 2 weeks after immunization, labeled stromal cells were microscopically found migrating directly into the joint cavity through the area between the articular margin and the synovial insertion (the bare zone). Labeled stromal cells were mainly found in the sublining layers of proliferating synovial tissue. CONCLUSION: At the onset of CIA, bone marrow stromal cells migrated from the bone marrow into the affected joint cavity and seemed to contribute to synovial proliferation.
Animals↗
Moloney murine leukemia virus infection accelerates lymphomagenesis in E mu-bcl-2 transgenic mice.
E mu-bcl-2 transgenic mice bearing the bcl-2 proto-oncogene linked to the immunoglobulin enhancer (E mu) sporadically develop B or T cell lymphomas after a long latent period. To identify genes that play important roles in development of lymphoid malignancies, proviral insertional mutagenesis with Moloney murine leukemia virus (MMuLV) was carried out in two lines of transgenic mice expressing the bcl-2 gene primarily in B or T cells. MMuLV infection of non-transgenic mice induced primarily mature T cell lymphomas. By contrast, infection of newborn E mu-blc-2 mice with the virus accelerated lymphomagenesis, and nearly all of the mice eventually succumbed to clonal pre-B, B, or mainly immature T cell lymphoma, indicating the active contribution of the bcl-2 gene in lymphomagenesis. Southern blot analysis of tumor DNA from MMuLV-infected transgenic mice revealed a proviral insertion at the c-myc gene in 26% (9/35) of tumors, at the pim-1 gene in 6% (2/35) and at the pim-2 (recently renamed tic-1) gene in 23% (8/35). Some tumors carried two activated oncogenes. No insertion was detected at the bmi-1 gene. These data suggest the usefulness of this transgenic system for analysis of lymphomagenesis involving the activated bcl-2 gene.
Aging↗