[Structures and functions of the renal tubules and interstitium].
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Animals↗
Biomedical subjects
S Inokuchi
Publications and source records attributed to S Inokuchi.
Rapid ex vivo expansion of human umbilical cord hematopoietic progenitors using a novel culture system.
Cell numbers limit the widespread clinical use of cord blood (CB) for gene therapy and marrow replacement in adults; a simple and effective method for ex vivo expansion of CB primitive progenitor cells (PPC) is required. Recently, the combination of thrombopoietin (TPO) and Flk-2/Flt-3 ligand (FL-2) was reported to support slow proliferation of CB-PPC in stroma-free liquid culture. We established a novel culture system in which the murine stromal cell line HESS-5 dramatically supports the rapid expansion of cryopreserved CB-PPC in synergy with TPO/FL-2. Furthermore, while HESS-5 cells directly adhered to human progenitors during culture, the cultured human cells could easily be harvested without contamination by HESS-5 cells. Within 7 days of culture, a 100-fold increase in CD34bright/CD38dim cells was obtained in serum-containing culture. When HESS-5 cells were physically separated from human progenitor cells in the presence of TPO/FL-2, synergy was blocked, suggesting that HESS-5 cells support proliferation of PPC by direct cell-to-cell interaction. The hematopoietic-supportive effects of this xenogeneic coculture system were then assessed in a very short-term (5 days) serum-free culture. Expansion was further enhanced by addition of stem cell factor (SCF) or interleukin-3 (IL-3). As a result, a 50- to 100-fold increase in CD34bright/CD38dim cells was noted. Colony-forming units in culture (CFU-C) and mixed colonies (CFU-GEMM) were enhanced by 10- to 30-fold and 10- to 20-fold, respectively. Moreover, generation of long-term-culture-initiating cells (LTC-IC) from CD34bright/CD38dim cells was amplified by 25-fold. The severe-combined immunodeficient (SCID) mouse-repopulating cell (SRC) assay confirmed extensive ability of the expanded cells to reconstitute long-term hematopoiesis. These results indicate that this xenogeneic coculture system, in combination with human cytokines, can rapidly generate PPC from cryopreserved CB.
Animals↗
The effects of tensile load on the metabolism of cultured chondrocytes.
The effect of cyclic tensile load on articular cartilage metabolism was investigated experimentally using 12 Japanese White rabbits. Chondrocytes obtained from the knee joints were cultured on plates with flexible silicone rubber bases. They were subjected to a cyclic (3 seconds on and 3 seconds off) tensile load for 24 hours with a maximum increase in area of 17%. Proteoglycan synthesis, collagen synthesis, and tissue inhibitors of metalloproteinases production by the chondrocytes under the load were quantified and compared with those produced by the control cells in an unloaded condition. The cultured chondrocytes under the cyclic tensile load perpendicularly aligned to the direction of the tensile load. Collagen synthesis and tissue inhibitors of metalloproteinases production increased significantly under the cyclic tensile load, although no significant change in proteoglycan synthesis was observed. These results suggested that the cyclic tensile load on the chondrocytes contribute to the regulation of articular cartilage metabolism in part.
Animals↗
Angiotensin II maintains the structure and function of glomerular mesangium via type 1a receptor. What we have learned from null mutant mice minus the angiotensin II type 1a receptor gene.
The angiotensin II type 1a (AT1a) receptor is the major receptor effecting the multiple actions of angiotensin II on the cardiovascular system. It is expressed abundantly in the glomerular mesangial cells of the kidney. We investigated glomerular changes in null mutant mice minus the AT1a receptor gene to gain an understanding of the in vivo action of angiotensin II via AT1a on the mesangium. Morphological observations and morphometric analysis revealed that the glomerular volume was greatly increased owing to the expansion of the mesangial area, which contained fluid-filled spaces with a small amount of fibrillar components. The mesangial cells lost contact with each other and with the perimesangial area of the glomerular basement membrane (GBM), so that the glomerular capillary neck was greatly widened. These findings suggest a defect of the anchoring function of mesangial cells resulting from some abnormality in mesangial matrix formation. We conclude that angiotensin II has an important role in the structural and functional maintenance of the mesangium via the AT1a receptor, especially by reinforcing the connection between mesangial cells and GBM via the mesangial matrix.
Angiotensin II↗
Histological and immunocytochemical studies of human psoriatic lesions transplanted onto SCID mice.
To investigate the pathology of psoriasis, we developed an animal model for this disease using severe combined immunodeficiency (SCID) mice. These mice possess neither B nor T Lymphocytes so that both cellular and humoral immunities are impaired. For the in vivo study of psoriasis, human psoriatic skin was grafted on SCID mice. Long-term morphological and immunohistochemical changes in the grafted skin ware examined for up to 22 weeks after transplantation. The human skin graft were generally well maintained during this period, but the histological and immunohistochemical findings characteristic of psoriasis, except for acanthosis and hyperkeratosis, gradually disappeared as lymphocytic infiltration of the psoriatic lesions declined.
Adult↗
The new-concept full-face-type helmet with removable cheek pads.
BACKGROUND: The American College of Surgeons proposed a method of removing helmets. But the problem with full-face-type helmets is that their shape makes them difficult to remove. METHODS: A dummy doll was fixed to a smooth bed surface in the supine position, and a full-face-type helmet with a hook attached to the vertex was placed on the doll's head. A spring balance was attached to the hook, traction was applied to the helmet through the spring balance, and the maximum tension needed to completely remove the helmet was measured. RESULTS: A tension of 13.2 +/- 1.8 kg was found. But when cheek pads were removed, the tension required to remove the helmet was 1.7 +/- 0.2 kg. CONCLUSION: We devised a full-face-type helmet that uses removable cheek pads so that helmet removal can be performed safely by removing only the cheek pads in the event of an accident.
Cervical Vertebrae↗
A simple and efficient purification of transduced cells by using green fluorescent protein gene as a selection marker.
BACKGROUND: Simple and efficient method for the selection of transduced cells would greatly facilitate the clinical utilization of retrovirus vectors. We developed a therapeutic bicistronic retrovirus vector for Gaucher disease, MFG-GC-GFP, which contains the human glucocerebrosidase (GC) gene and the green fluorescent protein (GFP) gene of the jellyfish Aequorea victoria as a vital selection marker, and investigated its applicability as gene therapy for Gaucher disease. METHODS AND RESULTS: A packaging cell line, GP + envAM12, was transfected with MFG-GC-GFP and, thus, produced a high titer recombinant virus (1.0 x 10(6) c.f.u./mL) in the culture supernatant. The expression level of GFP was correlated with the virus production in cells. The recombinant virus infected skin fibroblasts from a Gaucher patient and a sorted fraction of the cells expressing GFP by flow cytometry exhibited almost a six-fold higher activity of GC than normal fibroblasts. CONCLUSIONS: These data indicate that MFG-GC-GFP enables the one-step purification of a transduced fraction of target cells and is, therefore, considered to be a useful therapeutic vector for the experimental gene therapy of Gaucher disease.
Animals↗
Anterior subtalar dislocation: case report.
Anterior subtalar dislocations are extremely rare. To our knowledge, only four cases have been reported in detail in the literature. A diagnosis of anterior subtalar dislocation should be confirmed by an anteroposterior view radiograph because lateral subtalar dislocation always includes some anterior displacement of the mid-foot. We report a case of anterior subtalar dislocation confirmed by both lateral and anteroposterior view radiographs and discuss its pathomechanism, diagnosis, and treatment.
Adult↗
Posterior subtalar dislocation.
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Female↗
Closed complete rupture of the flexor hallucis longus tendon at the groove of the talus.
A rare case of closed complete rupture of the flexor hallucis longus tendon at its groove in the posterior process of the talus is reported in a soccer player who developed pseudarthrosis of the posterolateral tubercle of the talus after a Shepherd's fracture. Partial rupture or tenosynovitis of the flexor hallucis longus tendon at this level is well known in classical ballet dancers and soccer players. Three cases of complete rupture of the flexor hallucis longus tendon near the metatarsophalangeal joint and three under the sustentaculum tali have been reported, but there have been no reports at the groove of the talus. Repair was accomplished by tendon graft, and active flexion of the interphalangeal joint is now possible.
Adult↗
A kinematic study of ankle joint instability due to rupture of the lateral ligaments.
Kinematic analysis of patients who have ruptured the ligaments of the ankle joint was performed to evaluate the function of those ligaments. Ten patients with ruptured lateral collateral ligaments and 10 normal volunteers were examined. Patients' ages ranged from 17 to 29, averaging 20.9 years old. We performed kinematic evaluation by a three-dimensional optical analytic technique using surface markers. According to our results, the ankles with lateral ligament injury abnormally pronated and rotated externally at the time of heel strike and abnormally supinated and rotated internally during the acceleration phase.
Adolescent↗
[Umbilical cord blood transplantation for a patient with Diamond-Blackfan syndrome].
A five-year-old girl with Diamond-Blackfan syndrome received cord blood transplantation from an HLA-identical sibling. The patient showed pale face at birth, and was diagnosed to have Diamond-Blackfan syndrome. She had been treated with prednisolone (PSL), high dose of methylprednisolone, erythropoietin, and anti-lymphocyte globulin. Despite of these intensive therapies, erythropoiesis did not entirely improve, and transfusion of red blood cells had been required every third or fourth week until cord blood transplantation. Conditioning regimen consisted of thoraco-abdominal irradiation (TAI; 8 Gy), cyclophosphamide (CY; 50 mg/kg x 4), and anti-thymocyte globulin (ATG; 2.5 mg/kg x 4), Cyclosporin (CyA 3 mg/kg) was administered for the prophylaxis of graft-versus-host disease (GVHD). 4.14 x 10 (7)/kg of cord blood mononuclear cells were infused to the patient. White blood cell (WBC) and reticulocyte counts increased promptly, but recovery of platelet count was delayed. Skin GVHD (grade I) appeared on day +9, which responded to the administration of PSL (2 mg/kg). Chromosomal analyses of bone marrow cells for sex mismatch revealed complete chimerism on day +14, on day +28 and thereafter. Umbilical cord blood cells can be an alternative source of hematopoietic stem cells for allogeneic transplantation.
Child, Preschool↗
Location and action of angiotensin II type 1 receptor in the renal microcirculation.
To localize angiotensin II type 1a (AT-1a) receptor and to reveal the physiological roles of angiotensin II in the renal microcirculation, we investigated the AT-1a gene deficient mice, generated by a targeted replacement of the AT-1a receptor loci by the lacZ gene (Sugaya et al, J Biol Chem 270: 18719, 1995). Immunohistochemical localization of beta-galactosidase was performed in the heterozygous mutant mice to reveal the expression sites of AT-1a. The AT-1a receptor (that is, beta-galactosidase) was expressed both in the afferent and efferent arteriolar smooth muscles and also in the mesangial cells. The effect of angiotensin II on glomerular arterioles was directly observed using the hydronephrotic mice. Angiotensin II similarly constricted both the afferent and efferent arterioles in the wild-type and heterozygous mutant mice in a dose-dependent manner. This constriction was completely abolished by an AT-1 antagonist, CV-11974. In the homozygous null mutant mice, however, angiotensin II did not affect the arterioles at all. Electron microscopic studies revealed that the mesangial cells made contact with the glomerular basement membrane (GBM) at the capillary neck and also with each other in the wild-type mice. However, in the homozygous null mutant mice, the mesangial cells lost the contact either with GBM or with each other and thus the capillary neck became remarkably wider. The mesangial matrix area appeared loose and enlarged, suggesting impaired mesangial matrix formation. In conclusion, via the AT-1a receptor, angiotensin II equally constricts both the afferent and efferent arterioles and plays an essential role in maintaining the normal glomerular function and structure.
Angiotensin II↗
Construction and characterization of a recombinant adenovirus vector carrying the human preproinsulin gene under the control of the metallothionein gene promoter.
A new adenovirus vector carrying human-preproinsulin (h-PPI) genomic DNA, which was placed under the control of the mouse metallothionein gene promoter, was constructed. In the recombinant virus-infected cells, h-PPI gene expression increased as a function of ZnSO4 concentration. Reversed-phase high-performance liquid chromatography analysis revealed that the recombinant adenovirus-infected cells secreted immature insulin containing proinsulin and incorrectly processed insulin. Tyrosyl phosphorylation of human insulin receptor substrate 1 occurred when HepG2 cells were treated with the cultured medium, indicating that the h-PPI gene product was functionally active in vitro. We also examined the biological activity of the product using diabetic severe combined immunodeficient mice and confirmed that the h-PPI gene product reduced the blood glucose concentration in vivo. This study suggests that the adenovirus vector can be used to express a foreign gene under the control of an external promoter in various human cells.
Adenoviridae↗
Re-evaluation of foot process effacement in acute puromycin aminonucleoside nephrosis.
The sequence of morphological changes during foot process effacement in acute puromycin aminonucleoside (PAN) nephrosis was examined by means of NaOH maceration and freeze cracking for scanning electron microscopy (SEM). The micrographs of SEM and those of transmission electron microscopy (TEM) were quantitatively analyzed by computerized morphometry, and were correlated with renal function. On day 2 after PAN injection, the slit length was moderately decreased by both shortening and degradation of the foot processes. On day 4, membrane-bounded vesicles were scattered in the lamina rara externa. During foot process effacement, the basal surface of podocytes developed palm-like domains that represented the cytoplasmic areas between interdigitation. The decrease in the length of podocyte cell borders paralleled the decrease of 24-hour creatinine clearance. The development of the palm-like domains on the basal aspects of podocytes estimated by distance class analysis was closely correlated with the sudden onset of proteinuria. We conclude that foot process effacement in PAN nephrosis caused by the retraction and degradation of foot processes leads to the development of palm-like domains, which is correlated with podocyte detachment as well as massive proteinuria.
Animals↗
Muscle fibre composition and innervation ratios of the extensor digitorum and extensor hallucis brevis muscles in the crab-eating macaque (Macaca fascicularis).
The morphological and functional characteristics of the extensor digitorum and hallucis brevis muscles, the sole intrinsic muscles of the dorsum of the foot in the macaque, were investigated through dissection, the examination of muscle fibre composition, and counts of axon numbers. The total number of muscle fibers contained within the extensor digitorum (EHB) and extensor hallucis brevis (EDB) was almost 21,100. The population ratios of the three muscle fibre types of white, intermediate and red were 39.23 and 38%, respectively, for EDB and 40, 23 and 37%, respectively, for EHB. The mean cross-sectional area of each fibre type (white, intermediate, red) tended to be larger in the EHB (2,098, 1,480 and 911 microns2) than in the EDB (1,695, 1,310 and 822 microns2) and the value were significantly different for white fibres of males. The percentage area of the three muscle fibre types and the ratio of the white, intermediate and red areas, were similar between the EHB (55, 23 and 22%, respectively) and the EDB (52, 23 and 25%, respectively). The white area was dominant with the value showing a somewhat larger figure in the EHB compared to that of the EDB. The number of motor units was estimated to be 197-234 on average and the innervation ratio was calculated as between 91 and 109. These results suggest that the muscle fibre composition and innervation ratio of the EHB and EDB, especially the EHB, might relate to the arboreal locomotion of the crab-eating macaque.
Animals↗
Classification of fractures of the talus: clear differentiation between neck and body fractures.
The treatment and prognosis of neck fractures (extra-articular) and body fractures (intra-articular) of the talus are different. Ratios between neck fractures and body fractures reported by different investigators vary widely (from 6:1 to 1:1), because it is difficult to differentiate fractures crossing the anteromedial aspect of the trochlea. We examined 215 fractures of the talus. By examining the inferior surface fracture line, we found that the 61 fractures crossing the anteromedial aspect of the trochlea could be differentiated into 28 neck fractures and 33 body fractures. We suggest classifying fractures of the talus based on the inferior, not superior, surface fracture line.
Diagnosis, Differential↗
Bone loss of the radius in rheumatoid arthritis. Comparison between 34 patients and 40 controls.
We measured bone mineral density (BMD) in the radius by dual energy X-ray absorptiometry in 34 patients with rheumatoid arthritis (RA) and in 40 healthy controls. The BMD in RA patients in their fifties and sixties, but not in their forties and seventies, was significantly lower than that in the control subjects. The decrease in total radial BMD correlated with grip strength, RA activity and RA stage. The decrease in distal radial BMD correlated with RA activity, but not with grip strength. The levels of serum parathyroid hormone, alkaline phosphatase, and urinary hydroxyproline/creatinine were significantly higher in the patients. From these findings, we suggest that the bone loss in RA patients is affected by severity of inflammation, disuse, postmenopausal osteoporosis and secondary hyperparathyroidism.
Absorptiometry, Photon↗