Biomedical subjects
Publications and source records attributed to T Shiba.
We have isolated and characterized rainbow trout SOX9 cDNA and genomic clones. The cDNA encodes a protein of 488 amino acids (aa) with an HMG box and has 70% aa sequence identity with human SOX9. The rainbow trout and human SOX9 genes show a similar gene structure, and the two introns in the coding region are located at conserved positions. In rainbow trout, the SOX9 mRNA was prominent in testis and brain.
Inorganic polyphosphate [poly(P)] levels in Escherichia coli were reduced to barely detectable concentrations by expression of the plasmid-borne gene for a potent yeast exopolyphosphatase [poly(P)ase]. As a consequence, resistance to H2O2 was greatly diminished, particularly in katG (catalase HPI) mutants, implying a major role for the other catalase, the stationary-phase KatE (HPII), which is rpoS dependent. Resistance was restored to wild-type levels by complementation with plasmids expressing ppk, the gene for PPK [the polyphosphate kinase that generates poly(P)]. Induction of expression of both katE and rpoS (the stationary-phase sigma factor) was prevented in cells in which the poly(P)ase was overproduced. Inasmuch as this inhibition by poly(P)ase did not affect the levels of the stringent-response guanosine nucleotides (pppGpp and ppGpp) and in view of the capacity of additional rpoS expression to suppress the poly(P)ase inhibition of katE expression, a role is proposed for poly(P) in inducing the expression of rpoS.
We have isolated an SRY-type HMG box (Sox) cDNA, XSox3, from a Xenopus immature ovary cDNA library. The XSox3 cDNA contains an open reading frame (ORF) of 309 amino-acid residues, showing 62.7 and 77.0% homology with human and chicken Sox3 proteins, respectively. We also showed that the XSox3 gene is composed of a single exon by sequence analysis of the genomic clone and the determination of the transcription start site of the XSox3. The XSox3 mRNA was detected only in ovary and was at a higher level in immature ovary than in mature ovary. During oocyte maturation, the XSox3 mRNA was most abundant in stage I oocytes, and the XSox3 protein was detected in stage I and II oocytes. Recombinant XSox3 protein produced in Escherichia coli bound specifically to sequences containing the binding motif for the HMG box of SRY or SOX proteins, AACAAT or AACAAAG, demonstrating its sequence-specific DNA binding property. Taken together, these results indicate that the XSox3 protein may participate in early oogenesis of Xenopus as a transcription factor.
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GDP-L-Fuc:N-acetyl-beta-D-glucosaminide alpha1-->6fucosyltransferase (alpha1-6FucT; EC 2.4.1.68), which catalyzes the transfer of fucose from GDP-Fuc to N-linked type complex glycopeptides, was purified from a Triton X-100 extract of porcine brain microsomes. The purification procedures included sequential affinity chromatographies on GlcNAcbeta1-2Manalpha1-6(GlcNAcbeta1-2Manalpha1- 2)Manbeta1-4GlcNAcbet a1-4GlcNAc-Asn-Sepharose 4B and synthetic GDP-hexanolamine-Sepharose 4B columns. The enzyme was recovered in a 12% final yield with a 440, 000-fold increase in specific activity. SDS-polyacrylamide gel electrophoresis of the purified enzyme gave a major band corresponding to an apparent molecular mass of 58 kDa. The alpha1-6FucT has 575 amino acids and no putative N-glycosylation sites. The cDNA was cloned in to pSVK3 and was then transiently transfected into COS-1 cells. alpha1-6FucT activity was found to be high in the transfected cells, as compared with non- or mock-transfected cells. Northern blotting analyses of rat adult tissues showed that alpha1-6FucT was highly expressed in brain. No sequence homology was found with other previously cloned fucosyltransferases, but the enzyme appears to be a type II transmembrane protein like the other glycosyltransferases.
PURPOSE OF THE STUDY: To evaluate the effectiveness of testicular scintigraphy in differentiating testicular torsion from acute epididymitis. METHOD: 14 out of 17 patients previously screened by urologist were referred for testicular scintigraphy. Preoperative diagnosis was made based on dynamic and static image findings. Close follow-up was done and final diagnosis was confirmed on the basis of operative findings or relief of symptoms after conservative management. RESULTS: Eight of the nine patients with a photopenic area in the hemiscrotum were confirmed surgically as having testicular torsion. One patient was diagnosed as having an inflammatory process. Relief from symptoms after conservative management was noted in three patients with increased activity on the affected side and in two others whose uptake patterns were not different from the unaffected side. CONCLUSION: Testicular scintigraphy is effective in differentiating testicular torsion from acute inflammation when all other scrotal anomalies have been eliminated.
In spite of the routine use of hemoglobin Alc (HbAlc) value to guide diabetes therapy, substantial differences have been noted between results obtained by different instruments and laboratories. It was suggested that confusion between the terms used for HbA1c, i.e. between "total HbA1c' which includes labile content, and "stable HbA1c', which does not, may account for some of these inter-instrument and inter-laboratory differences. However, an undesirable difference was found even in measurements of only the stable component of HbAlc using the two most commonly used HPLC analyzers in our country. We found that a two-point calibration using lyophilized hemoglobin at lower and higher ranges was effective in matching up results of HbA1c measurement. This finding was concordant with a report by the Committee of the Japan Diabetes Society and other reports. Two methods of HbA1c immunoassay, one performed with the DCA 2000 automated analyzer and another performed using the Liquitech kit or a general automated analyzer, were evaluated and found to perform as well as HPLC analyzers in all respects except precision. The results derived from immunoassay were in good agreement with the calibrated results by HPLC, with (Liquitech) and without (DCA 2000) conversions.
BACKGROUND: Technical difficulties have impeded the development of laparoscopic hepatectomy. This article describes a new technique for performing partial hepatectomy and left lateral segmentectomy by means of laparoscopy, and the results in a series of 11 consecutive patients are reported. METHODS: A microwave tissue coagulator is used in combination with an ultrasonic surgical aspirator to divide hepatic parenchyma without pneumoperitoneum. Branched vessels and ducts are clipped and transected. The largest vessels were suture ligated in some cases. The endoscopic linear stapler was used to transect the left hepatic vein for left lateral segmentectomy. The resected liver was maneuvered into a specimen bag and removed. The argon beam coagulator was used to secure hemostasis of the plane of transection. RESULTS: Eleven patients underwent laparoscopic hepatic resection. Indications included isolated metastatic lesion, hepatocellular carcinoma, hemangioma, Wilson's disease, and hemochromatosis. Three patients underwent left lateral segmentectomy, and eight underwent partial hepatectomy. Ten procedures were performed uneventfully; one patient required conversion to open hepatectomy because of excessive bleeding. Notable differences were seen in blood loss compared with open hepatectomy, and no operative complications occurred. Postoperative pain was minimal. CONCLUSIONS: The laparoscopic hepatectomy, especially partial or left lateral segmentectomy, appears to be a viable surgical alternative in selected cases.
An assay method for GDP-L-Fuc:N-acetyl-beta-D-glucosaminide alpha 1-6fucosyltransferase (alpha 1-6FucT; EC 2.4.1.68) activity has been developed, involving a fluorescent pyridylaminated substrate. A glycopeptide derived from bovine gamma-globulin was coupled with 4-(2-pyridylamino)butylamine (PABA) through the peptide bond, and the following substrate was obtained. [equation: see text] The substrate and guanosine diphospho-fucopyranoside (GDP-Fuc) were incubated with a crude enzyme extract for 2 h, and then the enzymatic product was separated by reversed phase HPLC. Quantitation of the product involved measurement of the fluorescence intensity of the fucosylated pyridylaminated sugar. The structures of both synthesized GnGn-bi-Asn-PABA (substrate), and synthesized GnGnF-bi-Asn-PABA (product) were analyzed by 1H NMR. The enzymatic product was also analyzed by 1H NMR and was found to have alpha 1-6fucose at the reducing end GlcNAc. This method is highly specific for alpha 1-6FucT and is applicable for various experiments, including purification and cell culture ones.
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Fifteen (2.1%) patients were diagnosed as having congenital coronary artery fistula (CAF) in a consecutive series of 704 adult patients undergoing selective coronary arteriography; the incidence was astonishingly higher than previous observations. The presentation of clinical symptoms and the electrocardiographic changes at rest and/or after exercise testing probably attributable to the CAF were observed unexpectedly often in spite of the fact that the magnitude of the shunt seemed not to be significant. With respect to the anatomy of the CAF, the incidence of origination from plural coronary arteries and that of fistulation into the left ventricle were also unexpectedly high. These observations presented a striking contrast to those of previous reports, but we were unable to determine the reason. In four cases, the CAF was ligated either electively or concurrently with mitral valve surgery, and the results were satisfactory. Taking these circumstances into consideration, we should not minimize the impact of a CAF with a seemingly small shunt.
Hyperglycemia is the major causal factor in the development of diabetic vascular complications. The mechanism by which hyperglycemia causes the complications is not clear; however, it is very likely that hyperglycemia is mediating its adverse effects through multiple mechanisms. We have summarized some of these mechanisms in this review, with particular attention to the effect of hyperglycemia on the activation of diacylglycerol (DAG)-protein kinase C (PKC) pathway. We have reviewed existing information regarding various vascular tissues that show increased DAG and PKC levels. In addition, the mechanism by which hyperglycemia increases DAG as well as the cellular physiological consequences on the activation of PKC have been reviewed.
Renal cell carcinoma accompanied with ossification reported in Japan is very rare. We report a case of renal cell carcinoma with ossification pathologically in a 61-year-old woman. She suddenly presented with hypercalcemia that caused acute renal failure, 6 months after the operation and died shortly thereafter. We discuss the relationship between the tumor and the hypercalcemia.
A 68-year-old female presented ventricular septal perforation (VAP) 3 days after acute myocardial infarction. The patient, supported with intraaortic balloon pumping, underwent surgical repair with success on day 15. At operation, the VAP, measuring 2.5 x 1.5 cm, was repaired using a composite patch consisting of Dacron felt and xenopericardium attached each other with fibrin glue. The composite patch was found really useful for repair of VSP in such early phase as when VSP-sorrounding myocardium was very fragile, particularly in respect to the prevention of residual and/or recurrent shunt. This was because the composite patch was considered to have such excellent characteristics as: 1) toughness with sufficient compressibility and extensibility, 2) high antithrombogenesity and good tissue compatibility, 3) zero porosity, and 4) easy handling because of unification of two materials.
We assessed the Quality of Life (QOL) of 116 patients with inoperable esophageal cancer who were mainly treated with radiotherapy between 1978 and 1993. Factors including dysphagia, weight loss, side effects of radiotherapy,P.S. and psychological status were assessed retrospectively. Dysphagia was assessed using a swallowing-function scoring system. Initial improvement in dysphagia occurred in 68% of the 71 assessable patients. Radiotherapy was also effective for the other symptoms, including swallowing pain, anterior chest pain and vomiting. Improvement rates were 73%, 60% and 82%, respectively. The major side effects were pain of the pharynx, general fatigue, epigastralgia and appetite loss. But most side effects resolved within 10 days after the end of radiotherapy. Improvement of dysphagia resulted in reduced weight loss of the patients. Average weight loss was about 3 kg/month before radiotherapy. This was reduced after radiotherapy to 0.78 kg/month in patients who had improvement of dysphagia and 1.69 kg/month for those without improvement. Improvement of dysphagia also affected favorably the psychological state of the patients. There were fewer cases of patient distrust of staff and less despair in those patients with improved dysphagia. Radiotherapy appears to contribute to better QOL in patients with inoperable esophageal cancer.
Oral glucose tolerance test (OGTT) has been accepted as the standard test for screening of glucose intolerance in spite of its poor reproducibility, especially, in the impaired glucose intolerance (IGT). There have been widely assessed the diagnosis of diabetes mellitus with using glycemic control markers such as HbA1c, 1,5-AG, and FRA as an alternative method, which are not so much affected by sampling conditions. However, it has been found a wide overlap of the glycemic control marker's values between the normal and the IGT. In OGTT, subjects with lower blood glucose values 2 hours after glucose load turned out to be the normal glucose tolerance at the second examination as well. Therefore, it might be reasonable to identify people with a higher blood glucose value 2 hours after glucose load as a high risk group by the screening with fasting blood samples. We have evaluated the combination test of FBG, HbA1c and 1,5-AG for screening of the diabetes and the high risk IGT, and this combination revealed to be durable for screening. But it would be important to elucidate the natural history and the extent of the risk of the IGT, and select a proper test with a clear cut-off value for screening of impaired glucose intolerance at risk.
Ultrasound (US) hyperthermia is a local heating method that has recently been introduced into clinical practice. A prototype and modified version of this system using a phantom model (agar with bone inserted) were evaluated. The prototype system consists of a power delivery unit (0.5 MHz frequency) with a circulating cooled water bolus. This system heats a shallow area irrespective of power output, but has a tendency to overheat the phantom surface. We incorporated three modifications into the prototype. The modified type has three frequency applicators (0.5, 1.0, 1.5MHz) and a secondary bolus. Aluminum-urethane shields were used to cover the phantom surface located over bone preventing overheating of bone. By changing the applicator's US frequency, we could change the depth of the heating area. Overheating of the phantom surface and bone thus was avoided. These results suggest that our modified US hyperthermia system with multiple frequency applicators, a secondary bolus and aluminum-urethane shields provided a good heating pattern.