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Biomedical subjects

K Shinjo

Publications and source records attributed to K Shinjo.

At least 55 records · Page 3Linked to original sources
PubMed1997

Granulocyte colony-stimulating factor receptor at various differentiation stages of normal and leukemic hematopoietic cells.

Granulocyte colony-stimulating factor (G-CSF) is the most important cytokine in granulopoiesis, and induces the proliferation and differentiation of normal bone marrow granulocytic precursors. The physiologic effect of G-CSF is mediated through binding to specific cell surface receptors for G-CSF. Using a newly-devised quantitative flow-cytometric assay, we analyzed the expression of G-CSF receptors on normal and leukemic hematopoietic cells. In normal donors, G-CSF receptors are widely expressed from CD34-positive immature bone marrow cells to mature peripheral granulocytes. The highest expression of G-CSF receptors is observed in peripheral granulocytes. In the bone marrow, the level of G-CSF receptors expression increases in the following order; CD34+ CD33- cells < CD34+ CD33+ cells < CD34- CD33+ cells, indicating that G-CSF receptors are expressed on myeloid cells from a very early stage of differentiation and that the level of expression increases with the progress of cell maturation. G-CSF receptors are also detected on blast cells of patients with acute myeloid leukemia (AML) and acute lymphoid leukemia (ALL). The number of cell surface receptors varies from patient to patient, and no clear correlation is observed between the expression of receptors and the leukemia subtype or the cell surface markers. In this respect, the clinical use of G-CSF should be carefully controlled in ALL as well as AML patients.

Antigens, CD↗
PubMed1996

Sensitivity of opioid receptor-like receptor ORL1 for chemical modification on nociceptin, a naturally occurring nociceptive peptide.

Nociceptin or orphanin FQ is a novel neuropeptide that activates an opioid-like G protein-coupled receptor ORL1. This heptadecapeptide FGGFTGARKSARKLANQ resembles kappa-opioid peptide dynorphin A but exhibits an opposite effect to make animals hyperreactive to nociceptive stimulations (Meunier, J.-C., Mollereau, C., Toll, L., Suaudeau, C., Moisand, C., Alvinerie, P., Butour, J.-L., Guillemot, J.-C., Ferrara, P., Monsarrat, B., Mazarguil, H., Vassart, G., Parmentier, M., and Costentin, J. (1995) Nature 377, 532-535; Reinscheid, R. K., Nothacker, H.-P., Bourson, A., Ardati, A., Henningsen, R. A., Bunzow, J. R., Grandy, D. K., Langen, H., Monsma, F. J., Jr., and Civelli, O. (1995) Science 270, 792-794). In the present study, it was found that guinea pig brain contains receptors to which nociceptin binds much more strongly than to ORL1 receptors expressed in human 293 cells. Although the Tyr1 --> Phe substitution for dynorphin A eliminates almost completely an ability to bind to opioid receptors, the Phe1 --> Tyr substitution in nociceptin was found to retain almost fully both receptor binding affinity and in vivo hyperalgesic activity in tail-flick assay. Nociceptin was extremely weak to bind to opioid receptors, while Tyr1-nociceptin exhibited 10-40 times increased affinity, especially for mu receptors, due to its N-terminal sequential identity to opioid peptides. Shortened analogs of dynorphin A are known to retain receptor binding ability and analgesic activity, whereas the removal of C-terminal hexa- or decapeptide from nociceptin totally abolished the affinity for the ORL1 receptor. These results indicated that the mode of interaction between nociceptin and ORL1 receptor is quite different from that between dynorphin and opioid receptor and that the C-terminal portion of nociceptin is crucial for receptor recognition.

Amino Acid Sequence↗
PubMed1996

Expression of multidrug resistance P-glycoprotein in myeloid progenitor cells of different phenotype: comparison between normal bone marrow cells and leukaemia cells.

We examined the multidrug resistant P-glycoprotein (P-gp) on normal bone marrow (BM) cells and acute myeloid leukaemia (AMI) cells, using newly devised flow cytometric multi-parameter analysis with CD33, CD34 and MRK16 monoclonal antibodies. In both normal BM cells and AML cells, CD34+CD33- cells expressed P-gp strongly, CD34+CD33- cells moderately, and CD34-CD33+ cells weakly. Acute promyelocytic leukaemia, mainly expressing CD34-CD33+ but not CD34+CD33- at diagnosis, expressed less P-gp. P-gp expression of AML cells at diagnosis was increased as compared with normal cells of the same phenotype. P-gp expression was more increased in relapsed cases, especially in immature subpopulations.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1995

New flow cytometric method for detection of minimally expressed multidrug resistance P-glycoprotein on normal and acute leukemia cells using biotinylated MRK16 and streptavidin-RED670 conjugate.

To evaluate the expression of multidrug resistance (MDR) on normal and leukemia cells, we examined P-glycoprotein (P-gp) by a newly devised flow cytometric method, utilizing a biotinylated monoclonal antibody (mAb) against P-gp (MRK16), a streptavidin-RED670 conjugate (SA-RED670) and appropriate emission filters. The combination of biotinylated MRK16 (b-MRK16) and SA-RED670 resulted in higher sensitivity as compared with standard methods such as the use of streptavidin-phycoerythrin (SA-PE) conjugate. The sensitivity was examined in K562, K562/ADR, NOMO-1, NOMO-1/ADR and HL60 cells, and compared with the data obtained from reverse transcription polymerase chain reaction (RT-PCR) of mdr-1 gene. P-gp positivity on flow cytometry was 10.4%, 99.9%, 1.4%, 90.4% and 0%, respectively. Mdr-1 mRNA was well expressed in K562/ADR and NOMO-1/ADR cells, but not in NOMO-1 and HL60 cells. In K562 cells, mdr-1 was found after 40 cycles of PCR, but not 25 cycles. These data are well correlated with those from the flow cytometry. We then studied the P-gp expression on normal peripheral blood cells and acute leukemia cells. P-gp was little expressed on peripheral lymphocytes, monocytes and granulocytes. It was also little expressed on blast cells from 5 patients with acute promyelocytic leukemia (AML) and 5 acute lymphocytic leukemia (ALL) expressed P-gp at diagnosis, ranging from 8.5% to 34.5% (16.9 +/- 11.8%) and from 2.3% to 45.6% (24.0 +/- 17.8%), respectively. All 9 relapsed or refractory cases expressed P-gp, ranging from 21.1% to 99.8% (52.2 +/- 29.9%). Significant differences were found in APL, CD34-positive and relapse and refractory cases (P = 0.0006, 0.0007 and 0.0088, respectively). These results indicate that this flow cytometric analysis is useful for the evaluation of clinical MDR status and can identify a group of patients with resistant leukemia.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed1995

Expression of granulocyte colony-stimulating factor receptor increases with differentiation in myeloid cells by a newly-devised quantitative flow-cytometric assay.

In order to develop a non-isotopic quantitative assay of granulocyte colony-stimulating factor (G-CSF) receptors on human or murine cells, we devised a flow-cytometric assay using cells stained with biotin-labelled G-CSF (b-G-CSF) and a streptavidin-RED670 conjugate. For quantification, we applied the Kolmogorov-Smirnov test and calculated the D value. The D value was evaluated from the degree of shift in two fluorescence profiles according to the increase of fluorescence intensity due to the specific binding of b-G-CSF to G-CSF receptors. A good correlation was observed between the number of G-CSF receptors obtained by the radioisotopic binding assay and the number calculated from the D value by the flow-cytometric assay. Then, expression of G-CSF receptors on human bone marrow cells, peripheral blood granulocytes and blast cells from patients with acute myeloid leukaemia (AML) were studied. G-CSF receptors was expressed on CD34+CD33-, CD34+CD33+ and CD34-CD33+ cells in the following order: CD34-CD33+ > CD34+CD33+ > CD34+CD33- cells, indicating that the receptors increased with maturation. The receptor levels of CD34-CD33+ cells in bone marrow were apparently lower than those of CD34-CD33+ cells in peripheral blood granulocytes. On the other hand, an abnormal expression pattern of G-CSF receptors was noted in AML blast cells.

Acute Disease↗
PubMed1995

Allergic reaction involving liver dysfunction and disseminated intravascular coagulation caused by a health food, Proporis.

A 43-year-old woman was hospitalized with skin eruption and liver dysfunction complicated with disseminated intravascular coagulation (DIC) after taking Proporis, a so-called health food. Her clinical course was well correlated with discontinuation and the retaking of Proporis. No other reason for the development of DIC was detected except for allergic reaction to Proporis, as demonstrated by a patch skin test. DIC subsided after the discontinuation of Proporis and treatment with gabexate mesilate and heparin. The consumption of health foods with various ingredients is recently increasing. Thus, attention should be paid to any possible serious allergic reaction related to such foods.

Adult↗
PubMed1994

Analysis of prognostic factors and chemotherapy of malignant fibrous histiocytoma of soft tissue: a preliminary report.

The inter-group study of malignant fibrous histiocytoma of soft tissue (MFH) was performed during the period January, 1988, to August, 1992 (4 yr 7 mo). There were 107 patients evaluated with an age range of 11 to 92 yr and an average age of 62.0 yr. There were 63 males and 44 females. The duration of follow-up was from 7 mo to 4 yr 5 mo with an average of 2 yr 6.2 mo. MFH occurred principally as a mass on an extremity (lower extremity 62%), upper extremity 18%, trunk 25%, head and neck 2%). It typically involved deep fascia or skeletal muscle (68%) or superficial subcutis (35%). There were 77 storiform-pleomorphic, 22 myxoid, three giant cell, two inflammatory and three angiomatoid variants. The absolute three-year survival rate for all 107 patients was 72%. Patients with tumors located on extremities had a better three-year survival rate than patients with tumors located on the trunk or head and neck (81 vs 54%). There was, however, no statistical difference between the two groups. The prognosis for patients with deep muscular lesions, > 10 cm in diameter, was poor. The depth of the tumor significantly affects survival. The group with superficial tumors had a significantly better three-year survival rate than the other group (86 vs 68%). Local recurrences were found in 8% of patients who had had "complete" excisions. Inadequate surgery greatly increases the incidence of local recurrence and compromises longterm survival. Evaluations of tumor responses to preoperative chemotherapy and radiotherapy by histological studies were made. In total, 38 patients were evaluated. Responses to single and combination chemotherapy occurred in 31% of patients, a rate similar to that seen for other sarcomas.

Adolescent↗
PubMed1993

Biochemical comparisons of the Saccharomyces cerevisiae Bem2 and Bem3 proteins. Delineation of a limit Cdc42 GTPase-activating protein domain.

The Bem2 and Bem3 proteins, which appear to play roles in the regulation of bud site formation in Saccharomyces cerevisiae, show striking homology to a number of proteins that compose a family of GTPase-activating proteins (GAPs) for the rho-subgroup of ras-related GTP-binding proteins. These members include human platelet GAP for Cdc42Hs (the human homolog of a S. cerevisiae GTP-binding protein that regulates bud site assembly), the break point cluster region protein, the brain protein chimerin, the 85-kDa regulatory subunit (p85) of the phosphatidylinositol 3-kinase, and the ras-GAP-binding protein (p190). A fusion protein composed of the glutathione S-transferase protein and the rho-GAP homology region of Bem3 (designated GST-Bem3) stimulates the GTPase activity of the wild-type Cdc42Hs protein (Cdc42HsGly-12), but has no stimulatory effect on a GTPase-defective mutant (Cdc42HsVal-12), whereas a GST-Bem2 fusion protein does not stimulate the GTPase activity of either form of Cdc42Hs. We have compared the ability of GST-Bem3 to serve as a GAP for Cdc42Hs relative to other members of the rho-GAP subfamily and found the following order of potency: human platelet Cdc42Hs GAP > p190 > Bem3 > break point cluster region protein, whereas p85, like Bem2, shows no GAP activity or any ability to bind to the GTP-bound form of Cdc42Hs. We have taken advantage of the functional specificity exhibited by Bem3 (versus Bem2) in using Bem2/Bem3 chimeras, as well as different deletion mutant versions of the Bem3 protein, to delineate the limits of a functional Cdc42 GAP domain. The results of this study indicate that the carboxyl-terminal approximately 224 amino acids (which contain three regions of homology to the other members of the rho-GAP family) represent a "limit GAP." The first two appear to be important for binding to Cdc42Hs and for partial GAP activity.

Amino Acid Sequence↗
PubMed1993

Malignant giant cell tumor of the tendon sheath: an autopsy report and review of the literature.

A case of malignant giant cell tumor of the tendon sheath of the right hip, which developed in a 72-year-old Japanese woman, is described. The tumor exhibited histological similarities to a benign giant cell tumor of the tendon sheath (localized nodular tenosynovitis). The resected tumor, measuring 9 x 9 x 11 cm, was located in the adductor muscle and invaded the proximal femur and acetabulum. The nodule was encapsulated with a thin membrane which was soft and gelatinous in consistency and varied in color from yellow to brown. The synovium of the hip joint was normal. The primary lesion was composed of plump polyhedral and spindle-shaped cells. The nuclei were large, irregular and hyperchromatic, and contained prominent nucleoli. A moderated number of multinucleated giant cells was scattered throughout the lesion. There was little stromal collagen. In the majority of the specimens, pseudoglandular or alveolar spaces were predominant. An ultrastructural study demonstrated three cell types: fibroblast-like, histiocyte-like and an intermediate. The patient underwent reconstructive surgery with a Dacron fabric-enveloped alumina ceramic pelvic prosthesis and total hip components after resection of the primary lesion. Unfortunately, because of a local recurrence, a hemipelvectomy was required 10 months after the initial operation. At that time the intestines were involved with the recurrent tumor, and the patient subsequently died of perforative peritonitis. An autopsy revealed distant metastases to the right pelvis, urinary bladder, right ureter, ilium, mesenterium and lungs.

Aged↗
PubMed1993

[One stage operation for dead space of pyogenic and tuberculous osteomyelitis].

Pyogenic and tuberculous osteomyelitis is known to have difficult surgical problems. Complete surgical excision of dead and grossly infected soft tissues and osseous structures frequently results in large defects. Hydroxyapatite filler has been used to pack osteomyelitic defects. Between 1984 and 1991 we have treated 6 foci in 6 patients: 4 males aged 61, 54, 47 and 26 and; 2 females aged 59 and 36 years. There were 3 cases of chronic osteomyelitis, and one each of acute osteomyelitis, tuberculous osteomyelitis, and Brodie's abscess. All were reviewed within a period of between 21 to 94 months postoperatively. The pathogens cultivated were Mycobacterium tuberculosis in case 2 and Staphylococcus aureus in case 3 and case 6. In other cases, cultures revealed no pathogens. The hydroxyapatite fillers were mixed with an antibiotic. In addition, in the last four cases the fibrin sealant and CaCl2-thrombin solution were mixed. The excavated bone defect was packed with this composite biomaterial. Neither closed irrigation/suction using antibiotic solution nor a cast was used. Antimicrobial therapy directed specifically to the deep tissue specimens, was administered to all patients. On examination, all of the foci had completely healed by the end of the follow-up period. The cure of pyogenic and tuberculous osteomyelitis is obtained from obliterating dead spaces, which prevent the survival of organisms. Hydroxyapatite is the biomaterial most compatible with human bone and is also suitable for obliterating dead space. The advantages of the fibrin sealant system are that defects can be packed without gaps. Thus, postoperative hematomas can be avoided. In addition, the antibiotic incorporated into the fibrin sealant is released more slowly from the concentrated fibrin than from hematomas. As a result, pathogens multiply more slowly in fibrin. From these findings, we believe that this new method is simple, can be performed safely in one stage, and offers satisfactory results.

Adult↗
PubMed1993

[Reconstruction for malignant bone tumor defects by a composite graft--plasty of the bone defect not involving a joint].

Malignant bone tumors excluding joints of six patients were resected. The resultant bone defects were reconstructed with a synthetic fabric-enveloped hydroxyapatite spacer in five cases and a filler in one. Dacron fabric was used in four cases and each of Marlex mesh and Dexon mesh were used in one. The grafting and reconstruction processes were studied clinically and roentgenographically. CT scan was used in one case. The synthetic fabrics were helpful in repairing released supporting soft tissue by serving as a scaffold for the formation of new connective tissue. The spacer to the supporting soft tissues was biologically fixed. The synthetic fabrics were capable of inducing and supporting of the ingrowth of vascular connective tissue. The anchor of reconstructed supporting soft tissue became increasingly strong as time passed. For lasting and stable reconstruction, the spacer must be made uniform with the remaining bone by making an extracortical bone bridge using an autogenous bone graft. In two cases, autogenous bone grafts were taken from the ilium to bridge the remaining bone. The autogenous bone grafts were fixed to the spacer by enveloping the bone graft and the spacer with Marlex mesh and Dexon mesh. In the Marlex case roentgenographic findings showed that extracortical bone bridging was completed within six months after the operation. In the Dexon mesh case, extracortical bone bridging was not completed at one year after the operation, however the repair of the released supporting soft tissues was satisfactory. In order to avoid complications, a fabric must be absorbed fully after completion of the biologic fixation and extracortical bone bridging. Yet, further investigations are required to determine which of the synthetic fabrics and mesh sizes to enhance incorporation of the bone graft.

Adult↗
PubMed1993

Dacron fabric-enveloped alumina ceramic prosthesis for cementless resection arthroplasty of the proximal tibia: a case report with a long-term result.

After resection of the primary lesion, the right proximal tibia of a patient with osteosarcoma was reconstructed using a Dacron fabric-enveloped alumina ceramic total knee prosthesis without cement. Dacron fabric is intended to act as a scaffold upon which connective tissue can proliferate and form new ligament and other supporting soft tissues. Usually, following wide resection of the primary lesion, the supporting system is lost. If the supporting soft tissues are not reconstructed after removal of the tumor, they will be fixed in a slackened condition and will not provide sufficient support, leading to problems of stability and function. Using alumina ceramics alone does not induce surrounding connective tissue formation; therefore, stability is not ensured. The support of the vascular tissue enables the fabric to carry out the functions of ligaments, tendons, retinaculum, periosteum, joint capsule and other supporting soft tissues. Over time, stabilization occurs, biologic fixation of the prosthesis to the supporting soft tissues is successful and the durability of the prosthesis is prolonged. Postoperative follow-up has continued for a 77-month period. At present, the patient continues to show no evidence of disease and is able to walk painlessly up and down stairs without using a knee brace or handrail. A knee brace was recommended, however, to ensure the durability of the prosthesis. Dacron fabric is useful for the repair of supporting soft tissues without significant complications. Using this unique method, a stable, functional and aesthetically pleasing reconstruction can be accomplished.

Adult↗
PubMed1992

Reconstruction of a segmental clavicular defect with a Marlex mesh-enveloped composite graft consisting of a hydroxyapatite prosthesis and autogenous bone: a case report.

The present study reports a case of a 38-year-old man with a malignant lymphoma of the right clavicle who underwent reconstruction after a segmental resection of the clavicle. A hydroxyapatite prosthesis in the shape of the clavicle was placed across the defect in conjunction with Marlex mesh which entirely enveloped the prosthesis and iliac crest autogenous bone. The mesh also acted as a stabilizer to the segments. The postoperative management was not a burden to the patient. Postoperative radiographs indicated bony union to have been completed within six months of the operation. The patient returned to work as a driver eight months after surgery. The range of motion of the shoulder joint was not limited. The patient had no trouble in carrying out his daily life without pain. The appearance of the area operated was excellent. At the postoperative 39 months follow-up visit, a radiograph reconfirmed the stable reconstruction with a composite graft and bone bridging at the gap left in the prosthesis to have been maintained with little bone absorption. The results indicate a reconstruction using a Marlex mesh-enveloped composite graft consisting of a hydroxyapatite prosthesis and autogenous bone to be a useful alternative to other types of reconstruction for segmental clavicular defects.

Adult↗
PubMed1992

[Reconstruction for malignant bone tumor defects by a composite graft--cementless arthroplasty].

Five patients with malignant bone tumor were arthroplastied with Dacron fabric-enveloped prosthesis without using cement after resection of tumor. Usually following the wide resection of the primary lesion, the supporting tissues are always lost. Dacron fabric is intended to act as a scaffold upon which connective tissue can proliferate and form new ligament and supporting soft tissues. Dacron fabric is useful for the repair of ligament, muscles, joint capsule, periosteum, and other supporting soft tissues without significant complications. The lengths of the released supporting tissues are easily adjustable. Biologic fixation of the prosthesis to the soft supporting tissues is successful, then the durability of the prosthesis prolongs. The Dacron fabric is capable of inducing and supporting the ingrowth of vascular connective tissue which becomes stronger thereafter. The author believes that this method provides a stable, functional and esthetically pleasing reconstruction.

Adult↗
PubMed1991

Identification of the human platelet GTPase activating protein for the CDC42Hs protein.

The CDC42Hs protein appears to be an isoform of the ras-related GTP-binding protein G25K and is an apparent human homolog of the Saccharomyces cerevisiae cell-division-cycle protein, CDC42Sc. In this study, we report the identification of a GTPase-activating protein (GAP) for CDC42Hs from human platelets (designated from here on as CDC42Hs-GAP). The CDC42Hs-GAP activity was solubilized from platelet membranes, recovered through successive chromatography steps (the final step being Mono-Q chromatography), and purified approximately 3500-fold. The CDC42Hs-GAP activity appeared to correspond to a polypeptide with an apparent Mr of approximately 25,000. The GTPase activities of the purified human platelet CDC42Hs, the Escherichia coli-recombinant CDC42Hs, and the Spodoptera frugiperda-recombinant GTP-binding proteins are all stimulated by the CDC42Hs-GAP to identical extents, which indicates that the recombinant CDC42Hs proteins are as effective as the native human platelet protein in coupling to the GAP. However, a mutant form of the E. coli-recombinant CDC42Hs which contains a valine residue at position 12 (CDC42HsVal-12) has a significantly reduced intrinsic GTPase activity (relative to the wild type CDC42HsGly-12) which is not stimulated by the CDC42Hs-GAP. The CDC42Hs-GAP also does not stimulate the GTPase activities of the ras or rap GTP-binding proteins; however, it is capable of a weak stimulation of the GTPase activity of mammalian rho. Based on the apparent similarities in the molecular size of the CDC42Hs- and rho-GAPs (i.e. 25-30 kDa), and the cross-reactivity of rho with the CDC42Hs-GAP, it seems likely that the CDC42Hs- and rho-GAPs will constitute a specific subclass of the ras-related GAP superfamily.

Blood Platelets↗