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K Oshima

Publications and source records attributed to K Oshima.

At least 91 records · Page 5Linked to original sources

Bone morphogenetic protein 4 regulates the budding site and elongation of the mouse ureter.

In the normal mouse embryo, Bmp4 is expressed in mesenchymal cells surrounding the Wolffian duct (WD) and ureter stalk, whereas bone morphogenetic protein (BMP) type I receptor genes are transcribed either ubiquitously (Alk3) or exclusively in the WD and ureter epithelium (Alk6). Bmp4 heterozygous null mutant mice display, with high penetrance, abnormalities that mimic human congenital anomalies of the kidney and urinary tract (CAKUT), including hypo/dysplastic kidneys, hydroureter, ectopic ureterovesical (UV) junction, and double collecting system. Analysis of mutant embryos suggests that the kidney hypo/dysplasia results from reduced branching of the ureter, whereas the ectopic UV junction and double collecting system are due to ectopic ureteral budding from the WD and accessory budding from the main ureter, respectively. In the cultured metanephros deprived of sulfated glycosaminoglycans (S-GAGs), BMP4-loaded beads partially rescue growth and elongation of the ureter. By contrast, when S-GAGs synthesis is not inhibited, BMP4 beads inhibit ureter branching and expression of Wnt 11, a target of glial cell-derived neurotrophic factor signaling. Thus, Bmp4 has 2 functions in the early morphogenesis of the kidney and urinary tract. One is to inhibit ectopic budding from the WD or the ureter stalk by antagonizing inductive signals from the metanephric mesenchyme to the illegitimate sites on the WD. The other is to promote the elongation of the branching ureter within the metanephros, thereby promoting kidney morphogenesis.

Animals↗

An ELISA-based assay for detergent-solubilized cellular beta 1,4-galactosyltransferase activity. Use of a polyacrylamide derivative with GlcNAc-beta side chains as a solid phase acceptor substrate.

In our previous paper [Oubihi et al. (1998) Anal. Biochem., 257, 169-175], we have shown that a polyacrylamide-derived synthetic glycopolymer with GlcNAcbeta side chains, termed PAP(GlcNAcbeta), is useful as a solid phase acceptor substrate for the ELISA-based analyses of soluble beta1,4(-)galactosyltransferase (GalT) activity in milk. This method is now used to assay detergent-solubilized cellular GalT. The glycopolymer coated on polystyrene plates was shown to be highly stable against the non-ionic and ionic detergents tested (0 approximately 5% solutions of Triton X-100 and SDS). Such stability made it possible to incubate the ELISA plate with detergent-solubilized GalT and to wash the ELISA plate with SDS solution after the GalT reaction, leading to high accuracy and sensitivity of this assay. The GalT activity was assayed using this method for 1% Triton X-100 extracts of various tissue samples of mice and several cultured cell lines. The results showed that the specific GalT activity of tissue extracts was low in brain and intestine, and high in ovary, muscle, and kidney. As for the cultured cell lines, COS7, COMMA-1D and C2C12 cells showed high specific activity, while CHO and MDCK cells showed low activity. The myoblast C2C12 had a slight increase in GalT activity during starvation-induced cell differentiation. On the other hand, GaIT-I transcript estimated by RT-PCR rather decreased during C2C12 cell differentiation, suggesting a differentiation-dependent switch in GalT isozymes. Taken all together, the ELISA-based assay using PAP(GlcNAcbeta) as a solid phase acceptor substrate was demonstrated to be a useful method for the assay of membrane-bound galactosyltransferases.

Acrylic Resins↗

Microsatellite genotyping of post-PCR fluorescently labeled markers.

We show that a post-PCR multicolor fluorescence-labeling technique is applicable to multiplex microsatellite genotyping. Forty-three dinucleotide microsatellite markers, which are located on 11q13-23, a candidate region for dominant familial exudative vitreoretinopathy (FEVR), were used to evaluate the quality of the marker profile produced by this technique. Thirty-eight people from six families with this disease were subjects for genotyping. The samples revealed clearly separated fragment peaks with signal intensities sufficient for analysis. All genotypes were consistent with Mendelian inheritance within each family. This method is cost effective because it does not use expensive fluorescently labeled primers. It also has the advantage of avoiding ambiguity in the analysis, which may arise from the addition of non-template nucleotides by Taq DNA polymerase.

Fluorescence↗

Production and accumulation of thrombospondin-1 in human retinal pigment epithelial cells.

PURPOSE: To investigate the production and release of thrombospondin-1 (TSP-1), a natural inhibitor of angiogenesis, by human retinal pigment epithelial (RPE) cells to clarify the possible role of TSP-1 in maintaining intraocular angiogenesis. METHODS: Human RPE cells were isolated from a human cadaveric eye and cultured in medium with 5% newborn calf serum. TSP-1 messages in the purified RNA of RPE cells were analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Intracellular TSP-1 peptides were detected by cytofluorographic analysis. TSP-1 peptides in the culture medium on RPE cells were measured by sandwich enzyme-linked immunosorbent assay (ELISA). TSP-1 specific immunofluorescent staining was tested in RPE cells cultured on glass slides and in a human retinal tissue specimen. RESULTS: mRNA specific for TSP-1 was found in RT-PCR products from RPE cells, and it showed a time-dependent increase from the beginning of the culture. Intracellular staining for TSP-1 was identified by flow cytometry. The sandwich ELISA identified a time-dependent increase of TSP-1 peptides in the culture medium of RPE cells. Immunostaining for TSP-1 was observed in the cytoplasm of RPE cells cultured on glass slides. Positive immunostaining of TSP-1 was observed in the cytoplasm of the RPE layer in the human retinal tissue specimen. CONCLUSIONS: RPE cells can produce and release TSP-1 in vitro, and TSP-1 accumulates in the cytoplasm of RPE cells in vivo as well as in vitro. The production of TSP-1 by RPE cells is influenced by the state of proliferation and/or cell density. TSP-1 appears to be an important control factor in retinal and choroidal neovascularization.

Antibodies, Monoclonal↗

[New classification of the T category for pancreatic cancer].

The General Rules for the Study of Pancreatic Cancer (4th edition) is widely used in Japan, but several problems have been detected. One is the complexity of classification of the T category of peripancreatic invasion. This study was undertaken to simplify the classification of the T category. In 145 patients who underwent resection of pancreatic cancer, we analyzed the prognosis of those with invasion and without invasion based on seven factors of peripancreatic invasion. Furthermore, we selected three factors (S, invasion to the anterior pancreatic capsule; RP, invasion to the retroperitoneal tissue; and PL, invasion to the extrapancreatic nerve plexus) as prognostic factors by means of stepwise multivariate linear regression analysis. Subsequently we established a new classification of the T category: T1, without invasion in all three factors; T2, with invasion in only one of the three factors; and T3, with invasion in two or three factors. The prognoses of the same patients were analyzed retrospectively, and 1-, 3-, and 5-year survival rates were 100%, 88%, and 44% in T1, 76%, 30%, and 18% in T2, and 27%, 4%, 3% in T3 patients, respectively. In conclusion, we suggest that our new classification of the T category is simple and useful for the assessment of the prognosis in pancreatic cancer.

Female↗

Effect of scleral shortening on axial length.

BACKGROUND: Partial thickness sclerectomy is the most commonly employed scleral shortening technique used in conjunction with pars plana vitrectomy in the repair of myopia-associated macular holes in patients with staphyloma. Recently, scleral shortening induced through scleral invagination has been advocated as an adjunct in retinal translocation surgery. OBJECTIVE: To determine whether a correlation exists between the amount of sclera infolding and the posttreatment reduction in axial length (AL) as a result of lamellar scleral resectioning or full-thickness scleral invagination. METHODS: Three groups of 10 eyes each underwent lamellar scleral resection with dissection of 6, 8, and 10 mm in height, and 1 group of 10 eyes underwent a 10-mm invagination. Presurgical and postsurgical external AL of globes with stabilized intraocular pressure was measured to +/-0.022-mm precision. RESULTS: Average (+/-SD) AL shortening following lamellar resections for 6-, 8-, and 10-mm groups were 1.50+/-0.24, 2.10+/-0.13, and 2.65+/-0.24 mm, respectively, and 2.50+/-0.23 mm for the 10-mm invagination group. Differences in AL before and after scleral shortening were found to be significantly different between dissections of different heights (P<.05), and not significantly different between the 10-mm resection and invagination groups (P>.17). The AL of each group was shortened by approximately 25% of the resection-invagination height. The relation was quasilinear. CONCLUSIONS: Lamellar scleral resection and nonresected scleral invagination reduce the ocular AL. The extent of the reduction significantly correlates to the amount of removed or invaginated sclera. CLINICAL RELEVANCE: Surgical shortening of the sclera is useful in the management of several retinal disorders, but causes significant changes in AL. Arch Ophthalmol. 2000;118:965-968

Eye↗

Postoperative splanchnic perfusion following the reconstruction of thoracoabdominal aortic aneurysm involving abdominal visceral branches. Report of a case.

A 58-year-old man with thoracoabdominal aortic aneurysm involving visceral arteries underwent graft replacement of the thoracoabdominal aorta and associated reconstruction of abdominal visceral branches. Femoro-femoral extracorporeal bypass, intermittent selective visceral arterial perfusion and a staging cross-clamping method of the aorta were utilized at surgery. The total cross-clamping time of the aorta was 165 minutes. The patient's postoperative course was uneventful with no incidence of hepatic and renal failure or paraplegia. Although postoperative systemic blood pressure and cardiac output recovered to normal values within 7 hrs after the release of aortic clamping, it took 18 hrs for hepatic venous hemoglobin oxygen saturation (ShvO2) to recover. Base excess in arterial blood, lactic acid in arterial and hepatic venous blood, and ketone body ratio in arterial blood (AKBR) and hepatic venous blood (HVKBR) recovered to within normal ranges after 18 hrs of the release of aortic clamping. ShvO2 monitoring is a simple and may be a useful parameter in evaluating postoperative splanchnic perfusion and predicting abdominal organ failure at the time of thoracoabdominal aortic reconstruction involving visceral arteries.

Aortic Aneurysm, Abdominal↗

Relationship between Fas-ligand expression on carcinoma cell and cytotoxic T-lymphocyte response in lymphoepithelioma-like cancer of the stomach.

The Fas/Fas-ligand (Fas-L) system is involved in the induction of apoptosis and mediates T-cell cytotoxicity. We investigated the Fas/Fas-L system and cytotoxic T lymphocytes (CTLs) in 30 lymphoepithelioma-like cancer of the stomach (LECS) in order to understand the immune evasion of the tumor cells. Epstein-Barr virus (EBV) was detected in 15 cases in 30 LECSs. The expressions of Fas and Fas-L in tumor cells, and TIA-1, CD4, CD8 and CD56 in lymphocytes were examined by immunohistochemical staining. Apoptosis of tumor cells and lymphocytes was detected by the terminal deoxynucleotidyl-mediated dUTP-nick end labeling method (TUNEL). Expression of Fas and Fas-L was detected in tumor cells in 10 and 17 LECS, respectively. CTL consisted predominantly of CD8 (CD8(+) > CD4(+)), whereas natural killer (NK) cells were detected in 4 cases only. In Fas-L-positive tumors, the TIA-1-positive lymphocyte count was significantly lower (p < 0.05) and the number of apoptotic lymphocytes was significantly higher (p < 0.05) than in Fas-L-negative cases. The number of TIA-1-positive lymphocytes in EBV(+) cases was significantly higher than that in the EBV(-) tumors (p < 0.05). The number of apoptotic tumor cells in EBV(+) tumors was significantly lower than in EBV(-) cases (p < 0.01). Our results suggest that in LECS, tumor cells expressing Fas-L may evade the immune attack by killing lymphocytes through the Fas/Fas-L system. However, in EBV(+) LECS tumors, our results indicate that a high number of CTL is associated with a reduction in the number of apoptotic tumor cells. Our findings indicate that the Fas/Fas-L system plays a role in immune evasion of tumor cells in EBV(+) tumors. Int. J. Cancer (Pred. Oncol.) 84:339-343, 1999.

Aged↗

Lactation-dependent expression of an mRNA splice variant with an exon for a multiply O-glycosylated domain of mouse milk fat globule glycoprotein MFG-E8.

Expression of mRNA encoding MFG-E8, a milk fat-associated glycoprotein was investigated in mouse mammary gland. Two forms of mRNA, long and short variants, were shown to be expressed in the mammary tissue by RT-PCR analysis. Sequence analyses of these two variants and an isolated MFG-E8 gene segment indicated that the long and short mRNA variants resulted from an alternative splicing of a single pre-mRNA through in-flame inclusion and skipping of one exon, which encodes a proline/threonine (Pro/Thr)-rich domain. The long variant was expressed predominantly in mammary gland and the expression level was remarkably increased at late gestation and kept high during lactation. On the contrary, the short variant was detected ubiquitously in various tissues and its expression in the mammary gland was rather decreased in a lactation dependent manner. Expression of the long variant was also detected in a mouse mammary epithelial cell line, COMMA-1D, and enhanced by incubation with lactogenic hormones. Glycosylation inhibition analyses using tunicamycin and alpha-benzyl-GalNAc were conducted with COS7 cells transfected with plasmids expressing each mRNA variant, demonstrating that a fully glycosylated product of the long mRNA variant was not only N-glycosylated but also multiply O-glycosylated, whereas a product of the short one had only N-glycan(s). These results suggest that the alternative splicing plays a critical role for the mammary-specific and lactation-dependent expression of the MFG-E8 isoform and that the multiply O-glycosylated Pro/Thr-rich domain of this isoform is functionally important for formation of milk fat globules in mammary epithelial cells.

Alternative Splicing↗

The Effect of FR167653 on Cerebral Ischemia-Reperfusion Injury After Retrograde Cerebral Perfusion in a Canine Model.

Retrograde cerebral perfusion (RCP) has recently been reported to be useful for the repair of aortic arch aneurysms. However, there is a possibility that RCP supplies a limited amount of blood to the brain [1] and ischemia-reperfusion injury may occur after RCP. FR167653 (FR) is characterized as a potent suppressant of interleukin-1beta and tumor necrosis factor-alpha. We investigated the role of FR in preventing cerebral ischemia-reperfusion injury after RCP in a canine model. A total of 12 mongrel dogs was divided into two groups: in the FR group (n = 6), FR167653 (1 mg/kg/hour) was continuously administered during the period of RCP and rewarming; in the control group (n = 6), a physiological saline solution was administered at the same dosage as the FR167653 during the same period. Following hypothermia (20 degrees C) using cardiopulmonary bypass and circulatory arrest, RCP was performed by infusing oxygenated blood via the bilateral internal maxillary veins for 60 minutes at a perfusion pressure of 25 mmHg. The cerebral blood flow (CBF), cerebral metabolic rate for glucose (CMRGlu) and oxygen (CMRO2), and excretion of carbon dioxide (ExCO2) were measured. These results were expressed as the percentage of change from baseline values established immediately after anesthesia. CBF was significantly (p < 0.05) higher in the FR group than in the control group at 40 (159 +/- 25% and 82 +/- 21%, respectively) and 60 minutes (177 +/- 30% and 83 +/- 14%, respectively) after RCP. The lactate/pyruvate ratio of blood returned from the brain tissues was significantly (p < 0.05) lower in the FR group than in the control group at 40 and 60 minutes after RCP. CMRGlu was significantly (p < 0.05) higher in the FR group than in the control group 60 minutes after RCP. There was no significant difference in CMRO2 and ExCO2 between the two groups. It is concluded that FR167653 appears to be effective in protecting the brain from ischemia-reperfusion injury after RCP.http://link.springer-ny.com/link/service/journals/00547/bibs/8n3p143.html

Journal Article↗

The residual epiretinal membrane after vitrectomy for macular hole.

BACKGROUND: We retrospectively observed idiopathic macular holes in 63 eyes using a scanning laser ophthalmoscope, in order to study the relation between postoperative epiretinal membranes and closure of macular holes following vitrectomy. METHODS: The eyes were classified into three groups based on the degree of the postoperative epiretinal membranes. Group I consisted of 23 eyes with no epiretinal membrane remaining on the retina after vitrectomy. Group II consisted of 20 eyes in which epiretinal membranes were observed on the retina, but separate from the edge of the macular hole. Group III consisted of 20 eyes in which epiretinal membranes were observed at the edge of the macular hole. Using these three groups, we studied how postoperative epiretinal membranes were related to the closure of macular holes. RESULTS: All macular holes (100%) in groups I and II were closed following vitrectomy. In group III, 5 (25.0%) of 20 eyes had complete closure and 13 eyes (65.0%) had incomplete closure of the macular hole, while 2 eyes (10.0%) had re-opening of initially closed macular holes several months after vitrectomy. CONCLUSION: Residual postoperative epiretinal membranes at the edge of macular hole are responsible for primary failure of vitrectomy. Removal of epiretinal tissues around the macular hole is important for macular hole to be closed following vitrectomy.

Epiretinal Membrane↗

Cardioprotective effects of Lazaroid U-74389G on ischemia-reperfusion injury in canine hearts.

BACKGROUND: Lazaroid, an inhibitor of iron-mediated lipid peroxidation, has been shown to reduce free radical-mediated injury after ischemia and reperfusion. The effect of Lazaroid U-74389G was investigated on ischemia-reperfusion injury of the heart through preservation and transplantation (Tx) in dogs. METHODS: Eleven pairs of adult mongrel dogs weighing 8.5 to 12 kg formed the recipient-donor combinations. Following electromechanical arrest of the heart using cardioplegia, the coronary vascular beds were washed out with a cold University of Wisconsin solution followed by 12-hour preservation and orthotopic Tx. Experimental animals were divided into 2 groups; 6 pairs formed the control group, and 5 formed the Lazaroid-treated group in which Lazaroid U-74389G at 10 mg/kg was administered intravenously 30 minutes before reperfusion of the heart. The cardiac function including cardiac output, left ventricular (LV) pressure, and LV dp/dt was assessed 2 hours after Tx by comparing it with the recovery rates (%) from cardiac function of donor dogs. Each transplanted heart was harvested for histological study. RESULTS: The recovery of cardiac function after Tx was significantly better in the Lazaroid-treated group than in the control group. Histologically, myocardial damage, evaluated by both light and transmission electron microscopy, was less evident in the Lazaroid-treated group than in the control group. CONCLUSION: Early cardiac function following Tx was significantly better and histological damage was less in the Lazaroid-treated group than in the control group, suggesting that Lazaroid U-74389G is effective in preventing ischemia-reperfusion injury after preservation and Tx.

Adenosine↗

Long-term heart preservation using a new portable hypothermic perfusion apparatus.

OBJECTIVE: Perfusion storage is not often used clinically compared with simple immersion because of complicated circuits and demanding management. We developed a new apparatus for preservation combined with simple immersion and continuous coronary perfusion. METHODS: The main characteristics of this apparatus are as follows: (1) hypothermic storage, (2) does not require any energy source, (3) variable perfusion pressure, and (4) portability. The perfusion apparatus is composed of a storage chamber, a cooling chamber, and metal bars from which a perfusate bag is suspended. Adult mongrel dogs were divided into two groups: the coronary perfusion group (CP, n = 6) and the simple immersion group (SI, n = 6). Coronary vascular beds of the dog were washed out with a University of Wisconsin (UW) solution following cardiac arrest obtained using a GIK solution. The hearts were then excised. In the CP group, the heart graft, which was immersed in a 4 degrees C UW solution, was perfused with the same solution at a flow rate of 35 approximately 50 ml/hr. In the SI group, the heart graft was immersed in a 4 degrees C UW solution only. The heart graft was preserved for 12 hours in both groups. Beta-adenosine triphosphate (beta-ATP), phosphocreatine (Pcr), and inorganic phosphate (Pi) levels were measured immediately after excision of the heart, and at 3, 6, and 12 hours after preservation. Beta-ATP, Pcr, and Pi values were expressed as a percentage of control values, which had been obtained immediately after excision of the heart. Water content of the myocardium was measured prior to and after 12-hour preservation. The preserved graft was then evaluated through orthotopic transplantation. RESULTS: Beta-ATP/Pi levels at 6 and 12 hours after preservation were significantly higher in the CP group than in the SI group (62 +/- 5 versus 39 +/- 7%, 48 +/- 5 versus 22 +/- 8%, respectively, p < 0.05). Pcr/Pi levels at 6 and 12 hours after preservation were 30 +/- 9% and 22 +/- 8%, respectively in the CP group, while Pcr/Pi levels in the SI group were detected in only one case. There was no significant difference in water content either prior to or after 12-hour preservation between the two groups. Histopathologically, irregular expansion and/or contraction of myocardial fibers were more severe in the SI group than in the CP group. The recovery rate of hemodynamic parameters 2 hours after heart transplantation was significantly (p < 0.05) higher in the CP group than in the SI group. CONCLUSION: Stable and safe long-term canine heart preservation with continuous coronary perfusion associated with immersion is possible using this new apparatus, and may have broad clinical application.

Adenosine↗

A comparative study of Celsior and University of Wisconsin solutions based on 12-hr preservation followed by transplantation in canine models.

BACKGROUND: Celsior is a recently developed extracellular-type preservation solution that is effective in organ preservation. This experimental study was designed to compare the effects of Celsior and University of Wisconsin (UW) solutions in myocardial protection, using 12-hour preservation followed by orthotopic transplantation. METHODS: Fourteen pairs of adult mongrel dogs were divided into 2 groups. In the UW group (n = 7), UW solution at 4 degrees C was used for coronary vascular washout and storage following cardiac arrest with glucose-insulin-potassium (GIK) solution. In the Celsior group (n = 7), Celsior solution was used to produce cardiac arrest, for coronary vascular washout, and for storage. After 12-hour cold preservation, orthotopic transplantation was performed under cardiopulmonary bypass (CPB). The rate of recovery (%) of cardiac function of donor hearts was compared 1 and 2 hours after weaning from CPB, and then the transplanted hearts were harvested for histological study. RESULTS: Hemodynamic parameters including cardiac output, left ventricular pressure (LVP), and the maximum rates of positive and negative increase of LVP after transplantation were significantly (p < 0.05) higher in the Celsior group than in the UW group 2 hours after weaning from CPB. The transmission electron microscopic study found that degeneration of the mitochondria in the Celsior group was less extensive than in the UW group. CONCLUSION: Celsior solution enhanced the cardiac function of hearts preserved for 12 hours prior to transplantation compared to UW solution. Our results indicate that Celsior solution is equivalent or superior to UW solution for cardiac preservation.

Adenosine↗

The laminin-derived peptide YIGSR (Tyr-Ile-Gly-Ser-Arg) inhibits human pre-B leukaemic cell growth and dissemination to organs in SCID mice.

The YIGSR (Tyr-Ile-Gly-Ser-Arg) laminin beta1 chain sequence has an inhibitory effect on tumour growth and the metastasis of melanoma and fibrosarcoma cells. In the present study, we investigated whether the multimeric YIGSR peptide (Ac-Y16) has an antiproliferative effect and/or prevents the metastasis of human pre-B acute lymphoblastic leukaemia cells (NALM6) in severe combined immune deficient (SCID) mice. In in vitro studies, Ac-Y16 significantly inhibited leukaemic cell colony formation and the invasion of NALM6 cells in a Matrigel-based assay. The tumour growth and leukaemic infiltration in peripheral tissues were also analysed in SCID mice 9 weeks after NALM6, Matrigel and Ac-Y16 were subcutaneously co-injected. The weight of the subcutaneous tumours was significantly suppressed by Ac-Y16 in a dose-dependent manner. Flow cytometry analysis showed that the leukaemic infiltration was significantly inhibited in all organs with 1.5-2.0 mg of Ac-Y16. Leukaemic infiltrations in the brain were inhibited with 0.5 mg of Ac-Y16, and those in brain and bone marrow were also inhibited with 1.0 mg of Ac-Y16. With Ac-S16, a control-scrambled peptide, the only significant inhibition of the leukaemic infiltration was observed in bone marrow at a much higher dose. These data suggest that the multimeric YIGSR peptide can inhibit the tumour growth and metastasis of leukaemic cells and may be useful as a potential therapeutic reagent for leukaemic infiltrations.

Actins↗