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Biomedical subjects

S Sumi

Publications and source records attributed to S Sumi.

At least 37 records · Page 2Linked to original sources

Characterization of recombinant human chymase expressed in Escherichia coli.

We compared recombinant human chymase expressed in Escherichia coli with human chymase purified from vascular tissues. The recombinant chymase, the structure of which was NH2-enterokinase cleavage site-chymase-COOH, was expressed in Escherichia coli and then was solubilized and renatured. The protein did not have a chymase activity, but gained this activity after the cleavage of the N-terminal site by enterokinase. The enzyme was purified by heparin affinity and gel filtration columns. The N-terminal sequence of the protein was identical to the sequence for human chymase. The molecular weights of the recombinant chymase and chymase purified from human vascular tissues were 26 and 30 kDa, respectively, and the 4 kDa difference was thought to be due to the presence or absence of glycan. The optimum pH of the recombinant enzyme activity was between 7.5 and 9.0. The activity of the recombinant enzyme was inhibited by chymostatin, soybean trypsin inhibitor and phenylmethylsulfonyl fluoride, but not by ethylenediaminetetraacetic acid and aprotinin. This enzyme cleaved specifically the Phe8-His9 bond of angiotensin (Ang) I to form Ang II and that of big endothelin (ET)-1 to form ET-1-(1-31). These findings demonstrated that the enzymatic characteristics of the recombinant enzyme were identical to that of native human chymase.

Amino Acid Sequence↗

Superselective embolisation for intractable idiopathic epistaxis.

37 patients with intractable idiopathic epistaxis were treated with superselective embolisation between 1995 and 1999. A total of 40 embolisations was performed, including three procedures for recurrence. The embolic material was gelatin sponge in 27 procedures, microcoils in 9 and both gelatin sponge and microcoils in 4. Immediate cessation of nasal bleeding was obtained in all patients after embolisation. Recurrent epistaxis occurred in 2 (5.4%) of the 37 patients within 7 days after initial embolisation, giving a short-term success rate of 94.6%. The long-term follow-up ranged from 1-51 months (mean 21.6 months). Late re-bleeding occurred in two patients, giving a long-term success rate of 94.6%. Two patients underwent re-embolisation; it was necessary to embolise the ipsilateral facial artery and/or the contralateral internal maxillary as well as the ipsilateral maxillary artery. Although the overall complication rate was 45.0%, no major complications occurred. Superselective embolisation with gelatin sponge is an effective and safe treatment technique for intractable idiopathic epistaxis.

Adult↗

Screening for pyrimidine metabolism disorders using dried filter-paper urine samples: method development and a pilot study in Nagoya City, Japan.

A screening system for pyrimidine metabolism disorders by measurement with high-performance liquid chromatography using dried filter-paper urine samples is presented. This system permits the simultaneous determination of dihydrouracil, uracil, orotic acid and pseudouridine. The coefficient of variations for the four compounds on the filter-paper urine samples were 0.010 approximately 0.069 and the recoveries were 98.5 approximately 107.1%. The detection limits of the four compounds were 2 approximately 20 micromol/liter. The correlation between the filter-paper urine samples and liquid urine samples was excellent (0.938-0.988). We supeculated that this method could be used to detect pyrimidine metabolism disorders, such as dihydropyrimidinuria, dihydropyrimidine dehydrogenase deficiency and hereditary orotic aciduria. As a pilot study, we have analyzed dried filter-paper urine samples from 34, 200 healthy Japanese, and found three cases of dihydropyrimidinuria without clinical symptoms.

Drug Stability↗

Decreased fluorouracil cytotoxic effect on EB-virus transformed lymphocytes from hereditary orotic aciduria.

5-Fluorouracil (5-FU), used widely for malignancies, phosphorylate mostly by uracil phosphoribosyl transferase (UPRT). Patients with hereditary orotic aciduria lack the orotate phosphoribosyl transferase (OPRT) activity. In the cancer cells, the OPRT activity is paralleled with the UPRT activity. This study shows that the UPRT activity of the hereditary orotic aciduria homozygote decreased about 40% of normal controls. Moreover, we investigated the 5-FU cytotoxic effects on hereditary orotic aciduria (one homozygote, 4 heterozygotes and 7 normal controls), using EB-virus transformed lymphocytes (EB-LC). 5-FU was addded to the culture medium at concentrations ranging from 0 to 10.0 micromol/l. The 5-FU cytotoxic effects on the homozygote were milder than those on controls at each 5-FU concentration. The 5-FU cytotoxic effects in the heterozygotes were at intermediate levels between the homozygote and controls. We speculate that 5-FU cytotoxic effects, both anti-tumor effects and adverse reactions, would be weak when a patient with hereditary orotic aciduria was treated with 5-FU.

Antimetabolites↗

Detection of ornithine transcarbamylase deficiency heterozygotes by measuring of urinary uracil.

The importance of detecting heterozygosity for X-linked ornithine transcarbamylase deficiency is well known. Although the DNA analysis and the allopurinol loading tests are commonly used for this purpose, both methods require complicated procedures. In order to establish a simple test for detecting female heterozygotes, we examined the uracil and orotic acid in single-voided urine samples from 70 healthy women, and from 12 asymptomatic females with ornithine transcarbamylase deficiency. Based on the results of healthy women, we were able to determine a screening cut-off line of 11.9 micromol/mmol creatinine (mean +/- 1SD in logarithmic form) for uracil. Using this cut-off line, the sensitivity of OCT heterozygotes was 100%. We were also able to establish a second cut-off line of 28.9 micromol/mmol creatinine (mean +/- 3SD in logarithmic form) for diagnosis. Using this second cut-off line, the specificity of OCT heterozygotes was 100%. Our study has shown that the measurement of urinary uracil is a relatively simple and effective method for detecting female heterozygotes.

Female↗

Automated determination of 5-fluorouracil and its metabolite in urine by high-performance liquid chromatography with column switching.

We report a quantitative assay of 5-fluorouracil (FU) and its metabolite, 5-fluorodihydrouracil (FDHU) in human urine by used a column-switching high-performance liquid chromatographic method. The analyses were carried out using a molecular exclusion column for sample purification, and a cation-exchange column for separation. Each sample required only 40 min to analyze, and required no preparation other than filtration. Linearity was verified up to 1000 nmol/ml (r > 0.993). The recovery of FU was 96-101%; recovery of FDHU was 96-105%. The imprecision (RSD) for FU (10-100 nmol/ml) was < 1.5%, same-day (n = 5), and < 1.8%, day-to-day (n = 5). The imprecision (RSD) for FDHU (10-100 nmol/ml) was < 3.2%, same-day (n = 5), and < 4.0%, day-to-day (n = 5). The detection limits were, respectively, 0.1 nmol/ml. We measured FU and FDHU in urine of seven cancer patients after oral administration of FU. The cumulative quantity ratio of the FDHU and FU (FDHU/FU) excreted in their urine within 120 min after FU administration was a constant value in all seven patients. Based on these results, we believe that our method provides a useful tool for evaluating FU metabolism.

Breast Neoplasms↗

Serial lectin affinity chromatography demonstrates altered asparagine-linked sugar-chain structures of prostate-specific antigen in human prostate carcinoma.

Differences between prostate carcinoma (PCA) and benign prostatic hyperplasia (BPH) in asparagine (N)-linked sugar-chain structures of prostate-specific antigen (PSA) were investigated using serial lectin affinity chromatography. The amounts of PSA passing through columns of concanavalin A (Con A), phytohaemagglutinin E4 (PHA-E4) and PHA-L4 were significantly greater for PCA-derived PSA than BPH. We propose that the sugar moiety structure of PSA which is increased in PCA is a multiantennary complex type with branched N-acetylglucosamine beta(1->4) mannose. We suggest that N-linked sugar chains in PSA are altered during oncogenesis in the human prostate and may serve as diagnostic tools for PCA.

Aged↗

Alterations of asparagine-linked sugar chains of N-acetyl beta-D-hexosaminidase during human renal oncogenesis: a preliminary study using serial lectin affinity chromatography.

Enzymatic properties and asparagine (Asn)-linked sugar-chain structures of N-acetyl beta-D-hexosaminidase A (Hex A) were compared in human tissues between normal renal cortex and renal cell carcinoma (RCC). No significant differences between the two Hex A preparations were observed with respect to enzymatic properties such as molecular mass, Michaelis-Menten value or optimal pH. With RCC preparations, relatively more Hex A passed through the concanavalin A (Con A) column, bound weakly to Con A, or bound strongly to Con A and also to the wheat germ agglutinin (WGA) column, than with preparations from normal renal cortex. In contrast, relatively less Hex A bound strongly to the Con A column, but passed through the WGA column with RCC preparations than with those from normal renal cortex. Asn-linked sugar-chain structures might apparently be altered during human renal oncogenesis.

Asparagine↗

Complete nucleotide sequences of garlic viruses A and C, members of the newly ratified genus Allexivirus.

Complete genomic sequences of garlic viruses A (Gar V-A) and C (Gar V-C), members of an unassigned virus group recently identified in garlic plants, were determined. Their respective genomes consist of 8 660 and 8 405 nucleotides. The genomic structure and organization of these viruses are similar to shallot virus X (ShVX) which is the type species of the newly ratified genus Allexivirus. Phylogenetic analysis based on the amino acid sequences of the putative proteins including RNA-dependent RNA polymerase (RdRP), DNA helicase, or viral coat protein showed that the GarV-type viruses should be included in the genus Allexivirus. Furthermore, the amino acid sequence in the RdRP hypervariable region is highly divergent among the viruses in the genus Allexivirus, suggesting that they evolved independently from a hypothetical ancestor virus(es).

3' Untranslated Regions↗

Levels of free prostate-specific antigen (PSA) can be selectively measured by heat treatment of serum: free/total-PSA ratios improve detection of prostate carcinoma.

We studied a simple heat treatment method for measuring free prostate-specific antigen (PSA). Samples were incubated at 56, 58, and 60 degrees C for 5, 15, 30, 45, and 60 min. Then, 1 ml samples were fractionated on a Sephacryl S-200 gel filtration column to separate alpha1-antichymotrypsin-complexed PSA (ACT-PSA) and free PSA. Values of ACT-PSA decreased with increasing incubation temperature and time, whereas free-PSA remained relatively constant. The optimal temperature and time for incubation were 58 degrees C and 30 min. Using free/total-PSA ratios, we were able to distinguish between benign prostatic hyperplasia and prostate carcinoma in patients whose PSA was in the diagnostic 'grey zone', i.e. 4.1 to 10.0 ng/ml. Through receiver operating characteristic curve analysis, the area under the curve increased from 0.675 to 0.871 when comparing the performance of total PSA to the free/total-PSA ratio. Thus, clinical application of our present methodology may reduce the need to obtain prostatic biopsies in patients whose PSA level is within the diagnostic 'grey zone'.

Chromatography, Gel↗

Effects of intravenous administration of high dose-diethylstilbestrol diphosphate on serum hormonal levels in patients with hormone-refractory prostate cancer.

The objective of this study was to elucidate the mechanism underlying the further suppression of serum testosterone (T) by diethylstilbestrol diphosphate (DES-DP) in patients with prostate cancer refractory to hormonal treatment. These patients received an LHRH agonist with or without a non-steroidal androgen-receptor blocker or a gestagen before DES-DP. We measured serum levels of total and free T, dihydrotestosterone (DHT), estradiol (E2), dehydroepiandrosterone sulfate (DHEA-S), dehydroepiandrosterone (DHEA), androstenedione, cortisol, aldosterone before and during intravenous administration of high doses of DES-DP (500 or 1000 mg/day). DES-DP administration suppressed the serum levels of FSH (p=0.04) and total T (p=0.02), and eliminated free T (p=0.04) and E2 (p=0.04) from serum, while reducing serum DHEA-S to approximately two-thirds of the pretreatment level (p=0.03). In contrast, serum levels of SHBG (p=0.02) and cortisol (p=0.02) were markedly increased after DES-DP administration. The latter had no significant effect on serum levels of LH, DHT, ACTH, 17alpha-hydroxypregnenolone, 17alpha-hydroxyprogesterone, DHEA, androstenedione, or aldosterone. The results suggest that the potent suppression of circulating total T by DES-DP is caused, in part, by the inhibitory effect of DES-DP on serum DHEA-S level. In most patients, high-dose DES-DP treatment completely suppressed the serum level of free T, while possibly elevating serum SHBG and decreasing serum total T. The mechanisms that maintain the serum level of serum DHT during DES-DP treatment require further elucidation.

Aged↗

Power Doppler sonography assessment of tumor recurrence after chemoembolization therapy for hepatocellular carcinoma.

OBJECTIVE: This study was undertaken to determine the value of power Doppler sonography in the evaluation of recurrent hepatocellular carcinomas after transcatheter arterial chemoembolization therapy. SUBJECTS AND METHODS: Forty-five patients (age range, 45-81 years; mean age, 61 years) with hepatocellular carcinomas prospectively underwent power Doppler sonography, helical CT, and intraarterial digital subtraction angiography before and after transcatheter arterial chemoembolization therapy to evaluate for tumor recurrence. Three to 6 months after transcatheter arterial chemoembolization therapy, the Doppler signal, its location, and shape were evaluated. The results were compared with tumor vascularity as determined on helical CT and intraarterial digital subtraction angiography, which were used as the gold standards (n = 142). RESULTS: A sonographic signal was seen in 80 of 142 lesions with power Doppler sonography. Flow signal in lesions tended to be more difficult to detect in the left lobe (sensitivity, 74%) than in the right lobe (sensitivity, 93%). The location of the color signal was in the tumor's center, on its periphery, or both. No correlation between tumor recurrence and the location of a signal within a tumor was found. Power Doppler sonography had a sensitivity of 87%, a specificity of 85%, and an accuracy of 86% in revealing tumor recurrence after transcatheter arterial chemoembolization therapy. All lesions that showed linear signals in the center or at the periphery of the tumors proved to be recurring tumors (n = 20). In lesions with spotty signals, tumor recurrence was not revealed in nine of 60 lesions. CONCLUSION: Power Doppler sonography can be used for follow-up studies after transcatheter arterial chemoembolization therapy as a sensitive and cost-effective imaging technique. Although spotty signals were frequently seen, linear signals appeared to be specific for tumor recurrence.

Aged↗

[Long-term efficacy of the Gittes bladder neck suspension with use of ultrasonography for genuine stress urinary incontinence].

Thirty-eight patients with genuine stress incontinence underwent the Gittes procedures of the bladder neck suspension under ultrasonical monitoring. We performed the following two procedures. Original Gittes procedure: the puncture of the needle made twice through the different holes of the rectus fascia for each side and the bilateral helical suture was tied down separately above the rectus fascia. Modified Gittes procedure: the needle was passed through the rectus fascia once for each side and the both end of the helical suture was drawn up to suprapubic area, then the bilateral threads were tied over the rectus fascia. Original Gittes procedure was performed for twenty-nine patients and modified Gittes procedure for nine patients. Tightness of the suspension was decided by monitoring the posterior urethrovesical angle with use of the transrectal ultrasonography during the operation. A long term follow up survey was made up by means of a questionnaire by the phone or the mail. A total of thirty-six patients responded the questionnaire for a 94.7% response rate. The mean follow up was 33.1 months. 83.3% of the patients were cured and 5.6% were significantly improved. None of the patients claimed dysuria. There was no difference between the continent rates of the two procedures. We conclude that the Gittes bladder neck suspension with use of ultrasonography is effective for correction of female stress urinary incontinence.

Aged↗