Biomedical subjects

S Inokuchi

Publications and source records attributed to S Inokuchi.

At least 37 records · Page 2Linked to original sources

PubMed1997

[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↗

PubMed1997

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↗

PubMed1996

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↗

PubMed1996

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↗

PubMed1996

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↗

PubMed1996

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↗

PubMed1996

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↗

PubMed1996

Long-term follow up of talus fractures.

Eighty-six patients who had experienced a fracture of the talus more than 10 years previously were assessed. The fracture had occurred in the neck of the talus in 52 feet and in the body of the talus in 27 feet. The fracture was complicated by dislocation in 47 feet, and aseptic necrosis had developed in 33 feet. The outcome was rated as excellent in 20 feet, good in 43, fair in 18, and poor in 7. The cause of poor outcome in the late stage was posttraumatic osteoarthritis secondary to avascular necrosis and incongruity of the joint surface. Early active exercise without weight bearing is recommended to prevent contracture and bone atrophy due to disuse, which may promote osteoarthrosis. Arthrodesis is recommended in patients with pain because the outcome is greatly improved by arthrodesis, even when performed more than 10 years after the injury.

Adolescent↗

PubMed1995

Effects of fibroblasts of different origin on long term maintenance of xenotransplanted human epidermal keratinocytes in immunodeficient mice.

We examined effects of fibroblasts of different origin on long-term maintenance of xenotransplanted human epidermal keratinocytes. A suspension of cultured epidermal cells, originating from adult human trunk skin, was injected into double mutant immunodeficient (BALB/c nu/scid) mice subcutaneously, with or without cultured fibroblastic cells of different origin. At one week after transplantation, the epidermal cells generated epidermoid cysts consisting of human epidermis-like tissue. When the epidermal cells were injected alone or together with fibroblastic cells derived from human bone marrow, muscle fascia, or murine dermis, organized epidermoid cysts regressed within 6 weeks. In contrast, when the epidermal cells were injected together with human dermal fibroblasts, generated epidermoid cysts were maintained in vivo for more than 24 weeks. Histological examination showed that the reorganized epidermis, after injection of both epidermal keratinocytes and dermal fibroblasts, retained normal structures of the original epidermis during 6 to 24 weeks after transplantation. The results indicate that human dermal fibroblasts facilitate the long-term maintenance of the reorganized epidermis after xenotransplantation of cultured human epidermal keratinocytes by supporting self renewal of the human epidermal tissue in vivo.

Animals↗

PubMed1995

Establishment and characterization of a villous lymphoma cell line from splenic B-cell lymphoma.

A new B-cell line (VL51) with cytoplasmic villi was established from a female patient with splenic lymphoma with circulating villous lymphocytes (SLVL). The patient exhibited a clinical picture characteristic of SLVL, including massive enlargement of the spleen. Tartrate-resistant acid phosphatase (TRAP)-negative villous lymphocytes were seen in the peripheral blood, bone marrow (BM) and both red and white pulps of the spleen. Monoclonality of the VL51 cell line was confirmed by clonal genotype abnormalities in the immunoglobulin heavy chain (IgH) gene and the T-cell receptor beta (TCR beta) gene. Evidence for commitment of phenotype of the VL51 cell line to the B lineage was also shown by the immunophenotype, including expression of CD10, CD19, CD20 and surface immunoglobins. The VL51 cells were positive for Epstein-Barr virus nuclear antigen (EBNA). The VL51 cell line is the first SLVL cell line to be established, and it is expected to be useful in clarifying the leukemogenesis of SLVL.

Acid Phosphatase↗

PubMed1995

Immortalisation of human oesophageal epithelial cells by a recombinant SV40 adenovirus vector.

We introduced the origin-defective SV40 early gene into cultured human oesophageal epithelial cells by infection of a recombinant SV40 adenovirus vector. The virus-infected cells formed colonies 3-4 weeks after infection in medium containing fetal calf serum. When the cells derived from 'serum-resistant' colonies were then maintained in the serum-free medium with a low calcium ion concentration, some of them passed the cell crisis and kept growing for over 12 months. These cells, regarded as immortalised cells, resembled the primarily cultured oesophageal epithelial cells in morphology and had some of their original characteristics. Treatment of the cells with a high calcium concentration induced phenotypic changes. These cells still responded to transforming growth factor beta. When the immortalised cells were injected into severe combined immunodeficient mice, they transiently formed epithelial cysts, although the typical differentiation pattern of the oesophageal epithelium was not observed. These cysts regressed within 2 months without development into tumours. The results indicated that human oesophageal epithelial cells were reproducibly immortalised by infection with a recombinant SV40 adenovirus vector at relatively high efficiency. The immortalised cells should be useful in studies on oesophageal carcinogenesis and in assessing the cooperative effects with other oncogene products or carcinogens.

Animals↗

PubMed1994

Two adult cases of IgM-associated mesangial proliferative glomerulonephritis.

Two adult patients with mesangial proliferative glomerulonephritis with diffuse IgM deposition in the glomeruli are reported. Case 1 was a 25-year-old female with nephrotic syndrome who showed complete remission after treatment with prednisolone (PSL). Case 2 was a 46-year-old male with asymptomatic proteinuria who showed incomplete remission (0.5-1.0 g/24 hr) of urinary protein without any medication. In light microscopy, these patients revealed minimal or slight proliferation of glomerular mesangial cells without glomerular sclerosis and crescent formation. Deposition of IgM and C3 was observed in the glomerular mesangial areas and capillary walls by immunofluorescence. Electron-dense deposits were observed in the glomerular mesangial areas in these patients. Mesangial proliferative glomerulonephritis associated with diffuse IgM deposition in the glomeruli appears to have a benign clinical course. It has also been suggested that this disease has variant clinical courses since we recently experienced two other patients with mesangial proliferative glomerulonephritis with focal IgM deposits who showed renal tubular dysfunction or chronic renal failure.

Adult↗

PubMed1994

[A case report of left limb compartment syndrome associated with laparoscopic surgery].

A 63-year-old male patient with early gastric cancer was attempted for laparoscopic wedge resection of the stomach. After the introduction of anesthesia, bandage of bilateral leg was carried out to prevent deep venous thrombosis during laparoscopic surgery. Although the procedure was converted to open surgery, the bandage has been continued throughout the surgery for 6 hours. After the operation, the swelling and severe tenderness at his left leg was observed. MRI revealed remarkable edema in left deep posterior compartment. Under diagnosis of left limb compartment syndrome, fasciotomy was carried out. Postoperatively the patient did well without any functional disturbance.

Bandages↗

PubMed1994

Hematopoietic supportive function of human bone marrow stromal cell lines established by a recombinant SV40-adenovirus vector.

We have previously reported the establishment of a variety of human bone marrow stromal cell lines using a recombinant SV40-adenovirus vector. Using this vector, we obtained more clonal stromal cells. Here, we have characterized these cells and analyzed their capacity to support the proliferation and differentiation of hematopoietic cells. The stromal cells were cocultured with nonadherent human bone marrow cells used as hematopoietic cells. The total numbers of hematopoietic cells and CFU-GM in culture were counted every week. Two of the six stromal cell lines, AA101 and HAS303, supported the proliferation and differentiation of hematopoietic cells and CFU-GM for more than 9 weeks. Further, granulocytes, macrophages, and megakaryocytes were detected when cocultured with these cells. When hematopoietic cells were cocultured but separated from the two stromal cell lines by a 0.45-microns millipore membrane to prevent their attachment, almost all CFU-GM disappeared within 7 weeks. The supportive stromal cells produced GM-CSF and IL-6. However, other cell lines producing these humoral factors did not support hematopoietic cell proliferation for such a long time. These findings suggest that these established human bone marrow stromal cell lines will be useful, in that analysis of their supportive function in hematopoietic cell proliferation and differentiation through cell-to-cell interaction will shed some light on this area.

Adenoviridae↗

PubMed1993

Ultrastructural changes in glomerular epithelial cells in acute puromycin aminonucleoside nephrosis: a study by high-resolution scanning electron microscopy.

Ultrastructural changes in the podocytes were studied during the development of, and recovery from, acute puromycin aminonucleoside (PAN) nephrosis using high-resolution scanning electron microscopy (hrSEM) and transmission electron microscopy (TEM). In the process of development of PAN nephrosis, four types of early structural changes were observed before total loss of foot processes: formation of cytoplasmic blebs, masking of filtration clefts, flattening of foot processes, and retraction of foot processes. The masking of filtration clefts visualized by hrSEM corresponded to the multiplication of slit diaphragms and adhesion of foot processes in the TEM findings, and preceded retraction of the foot processes. Changes of podocyte configuration were produced. Recovery from this change of podocyte configuration began as islands of podocyte interdigitation, and was proceeded by expansion of the islands. During recovery, the primary processes were re-established either by retraction or perforation of the thin cytoplasm after the formation of foot processes. We conclude that loss of foot processes begins with the masking of filtration clefts. Recovery from the change in podocyte configuration begins with the formation of new foot processes.

Acute Disease↗

PubMed1993

Diffuse proliferative glomerulonephritis with hepatitis C virus-like particles in paramesangial dense deposits in a patient with chronic hepatitis C virus hepatitis.

A 62-year-old man with hepatitis C virus (HCV) infection developed proliferative glomerulonephritis with IgM and C3 deposits. Electron microscopy showed HCV-like particles in the paramesangial dense deposits, which are similar in shape to HCV previously described. These findings suggest HCV-related proliferative glomerulonephritis.

Chronic Disease↗

PubMed1993

Use of cultured human epidermal allografts for the treatment of extensive partial thickness scald burn in children.

In spite of recent progress in burn treatment, the early surgical therapy of partial thickness scald burns in children is still controversial. Early tangential excisions is not easily applicable for these patients because of difficulties in determination of the burn depth and probable physiological derangement after surgery. Hypertrophic scar formation and wound contraction after meshed autografts are other limitations. For these reasons, conservative treatment, not early excision therapy, has been chosen initially for these injuries. We used cultured epidermal allografts for extensive, partial thickness scald burns, during the early post-burn period without escharectomy. Fifty to 100% of the engrafted superficial dermal burns were epithelialized within 7 days. In contrast, untreated identical wounds remained open. Repeated grafting of cultured allografts on unexcised wound granulations of dermal burns also enhanced epithelialization. Long term results showed that hypertrophic scar formation in the mixed superficial and deep dermal burns was reduced when cultured allografts were used. Allografting of the cultured epidermis without surgical excision apparently promoted the rapid regeneration of the partial thickness burns. Procedural complications did not occur. Cultured allografts should be used as an effective and safe biological dressing for partial thickness scald burns in children.

Burns↗

PubMed1993

A case of nephrotic syndrome associated with hepatic glomerulosclerosis and diabetic nephropathy.

We report here an adult case of nephrotic syndrome associated with hepatic glomerulosclerosis with the hepatitis C virus (HCV) antigen and diabetic nephropathy. To clarify the etiology of the nephrotic syndrome, we performed a renal biopsy and obtained the histological findings of hepatic glomerulosclerosis, glomerular mesangial cell proliferation and mesangial expansion. Electron dense deposits and deposits of IgA and C3 were also noted in the glomerular mesangial areas. Histological findings of diabetic nephropathy included thickening of the glomerular basement membrane and aneurysmal change of glomerular capillary walls in light microscopy. In immunofluorescence, linear staining of IgG was observed in the glomerular capillary walls, Bowman's capsules and tubular basement membranes. HCV was also detected focally in the glomerular capillary walls by immunofluorescence. Association of these two diseases has not been reported in any of the previous manuscripts that we reviewed. Thus, this patient seems to be a case of very rare association of the two diseases.

Adult↗