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Biomedical subjects

S Tsuchida

Publications and source records attributed to S Tsuchida.

At least 19 recordsLinked to original sources

Molecular genetic basis of red cell markers and its forensic application.

(1) The polymerase chain reaction (PCR) was used to amplify Rh-related cDNAs from erythroid cells cultured by the selective two-phase liquid culture system for human erythroid progenitors in peripheral blood. Two Rh polypeptide cDNAs have been isolated from the PCR products and tentatively designated RhPI cDNA and RhPII cDNA. Both cDNA clones have an open reading frame composed of 1251 nucleotides. The RhPI cDNA clone shows a single nucleotide substitution with no amino acid substitution compared with the published sequence. The RhPII cDNA clone differs from the above by 41 nucleotide substitutions in the open reading frame, resulting in 31 amino acid substitutions. Besides these cDNA clones, eleven and five truncated isoforms of the RhPI and RhPII cDNAs, have been isolated, respectively. (2) The promoter region of the Duffy gene was cloned by IPCR of 1.1 kb SacI fragment and the 3' flanking sequence was cloned by IPCR of 1.9 kb EcoRI fragment. The IPCR products contained the known Duffy cDNA sequence without introns. By comparing the coding area of the Duffy gene in 28 Duffy positive individuals, we elucidated that one base change that results in an amino acid substitution (GAT(Asp44)-->GGT(Gly)) is in accordance with the Fya/Fyb polymorphism. This fact proves that the Duffy cDNA and its gene encode the Duffy blood group system. (3) Two common alleles in Esterase D (EsD) polymorphism, EsD1 and EsD2 were characterized by the substitution of one amino acid (Gly-Glu) caused by the point mutation of one nucleotide (G-A). The point mutation between cDNAs of EsD1 and EsD2 alleles was detectable as restriction fragment length polymorphism (RFLP) using Ssp1. The RFLP makes it possible to determine the EsD phenotypes using DNA samples from forensic materials without EsD enzymatic activity. (4) The authors report studies on 19 pairs of donors and recipients in bone marrow transplantation. A broad range of genetic markers at 42 gene loci, including one DNA marker 11 red blood cell markers, five human lymphocyte antigen types, 12 serum protein markers, five red cell enzyme markers, and eight salivary markers was evaluated before and after BMT over about 2 months. As a result, 11 out of 42 gene loci of genetic markers in recipients were transformed into the donor type.

Base Sequence

Stimulatory and inhibitory effects of sodium nitroprusside on soluble guanylate cyclase.

We investigated, using rat brain cortex supernatant as a source of guanylate cyclase (GC), whether sodium nitroprusside (SNP) can not only activate but also inhibit GC. SNP (I and 10 microM) activated the rat brain GC; however, at higher concentrations GC activation was reduced, resulting in a bell-shaped concentration-activation curve. Preincubation of GC with 10 microM SNP attenuated GC activation by SNP, S-nitroso-N-acetylpenicillamine (SNAP) or 3-morpholinosydnonimine-N-ethyl-carbamine (SIN). Such inhibitory effects of SNP were partially supressed by a nitric oxide (NO) scavenger oxyhemoglobin. The preincubation of GC with K4Fe(CN)6 (a carrier molecule for SNP but devoid of NO) had no inhibitory effects on GC activation. These results indicate that SNP, probably through NO, has dual effects on GC activity, stimulation and inhibition.

Animals

Ultrasonography for the detection of ureteric reflux in infants with urinary infection.

Several less harmful methods than voiding cysto-urethrography for detecting significant ureteric reflux have been proposed. The present prospective study investigated whether ultrasonography was effective in identifying ureteric reflux in infants with their first febrile urinary infection. The subjects were 27 infants (24 boys and 3 girls) aged from 0 to 8 months. The urinary tract was scanned when the bladder was full, and before and during induced voiding. Infants with abnormal ultrasound findings underwent voiding cysto-urethrography. The other infants were followed and those who had a recurrence of urinary infection underwent voiding cystography. Ten children underwent cysto-urethrography, with eight refluxing ureters identified in six boys. Ultrasound revealed transient dilatation of the renal pelvis on voiding in five kidneys, transient dilatation of distal ureters in 12 and hydro-ureteronephrosis in two. Each of the five kidneys with pelvic dilatation on voiding was associated with ureteric reflux grades III or IV. Of the 17 children who did not undergo cysto-urethrography, only one had recurrence of urinary infection and was diagnosed with ureteric reflux. This girl was one of the three babies who were not scanned during voiding. More than half of the infants with febrile urinary infection were excluded from invasive examination without having recurrence of urinary infection. Thus, ultrasound scanning during voiding was effective for screening infants with their first urinary infection to detect significant ureteric reflux.

Diagnosis, Differential

Hyporesponsiveness to nitrovasodilators in rat aorta incubated with endotoxin and L-arginine.

We studied the effect of L-arginine on relaxation responses to sodium nitroprusside or SIN-1 (3-morpholinosydonimine-N-ethyl-carbamine) in the rat thoracic aorta incubated with endotoxin. Sodium nitroprusside or SIN-1 produced a reproducible relaxation in the aorta incubated for 12 h with endotoxin. However, the response to both nitrovasodilators was remarkably attenuated when the aorta was preincubated for 12 h with endotoxin and L-arginine. D-Arginine could not substitute for L-arginine. The attenuated response to sodium nitroprusside or SIN-1 was partially restored by the inhibition of nitric oxide (NO) production with N omega-nitro-L-arginine. Cycloheximide prevented the inhibitory effect of preincubation with L-arginine. These results suggest that the prolonged exposure to muscle-derived NO induces hyporesponsiveness to nitrovasodilators.

Animals

Acute tubulointerstitial nephritis in association with Yersinia pseudotuberculosis infection.

A 4-year-old girl was diagnosed as having acute renal failure due to tubulointerstitial nephritis. The girl presented with remittent fever, vomiting and non-oliguric acute renal failure with sterile pyuria and tubular reabsorptive dysfunction. Ultrasound examination revealed that the kidneys were markedly enlarged with diffuse hyperechogenicity in the cortex when the abnormal renal function was present and were restored in size and echogenicity when the renal function normalised. A diagnosis of Yersinia pseudotuberculosis infection was based on a rise in haemagglutination titres against the organism.

Acute Disease

Ultrasonographic evaluation of bladder volume in young children.

We investigated by ultrasonography the bladders of 30 young children (26 boys, 4 girls) without established bladder control immediately before voiding cystourethrography. The anterior/posterior (D), superior/inferior (H) and transverse dimension (W) of the bladder were determined. The small children had a significantly greater H value compared with D and W than older children in our previous study in 1993. The regression equation for D x H x W and actual bladder volume in 30 infants was calculated as y = 0.49 x + 3 (Eq. 1). When bladder volume of the 30 infants was estimated from D x H x W, the mean percentage error was much lower with Eq. 1 than with Eq. 2 (y = 0.68 x + 4), which had been determined for older children (mean +/- SD 16.8% +/- 10.7% vs. 44.7% +/- 26.2%) (P < 0.01). Thus, infants have a different bladder shape than older children and their bladder volume should be estimated using a specific formula designed for them.

Child, Preschool

Lighted mirror for microneurosurgery.

We describe a newly developed, lighted mirror that provides enough luminous intensity to obtain a clear image in deep operative fields under high magnification. Through the initial neurosurgical procedures in which it was tested, the mirror proved particularly useful for inspecting the ventral aspect of the fifth nerve in a microvascular decompression for trigeminal neuralgia, and for observing the fundus of the internal auditory meatus after removal of an acoustic neuroma to ensure no residual tumor.

Humans

Surgery for cavoatrial extension of malignant tumors.

Surgical management for cavoatrial involvement of malignant tumors and its outcome is reported on for 6 patients; their age ranged from 55 to 79 years and 5 were male and 1 female. The basic disease was renal cell carcinoma in 5 cases and adrenal leiomyosarcoma in 1. Intracaval tumor extension was diagnosed by computed tomography, magnet resonance imaging, digital subtraction angiography, and echocardiography. The tumor was resected together with adherant vena cava and invaded right-atrial wall, using cardiopulmonary bypass and normo- or mild-hypothermia in 5 patients. The caval defect needed to be reconstructed with a slit GORE-TEX vascular prosthesis in 3 patients. In all patients the tumor resections were successful and without major complications. All patients survived and are well from 4 to 52 months after the surgery. It is concluded that such cavoatrial extensions of malignant tumors can be safely and accurately resected with the aid of cardiopulmonary bypass, with favorable early and late outcomes in patients who have no distant metastatic lesions.

Adrenal Gland Neoplasms

Lack of correlated expression between the glutathione S-transferase P-form and the oncogene products c-Jun and c-Fos in rat tissues and preneoplastic hepatic foci.

Since the expression of glutathione S-transferase P-form (GST-P) has been suggested from in vitro studies to be partly regulated by the oncogene product, c-Jun and c-Fos, their distributions were compared in normal rat tissues and preneoplastic hepatic lesions induced by the Solt-Farber protocol. Immunohistochemically demonstrated GST-P protein was positively correlated with expression of both c-Jun and c-Fos in the epidermis of the skin and the smooth muscle of adult lung and with either c-Jun or c-Fos respectively in the bile ducts and bronchial epithelium. However, GST-P expression was also observed in proximal and distal straight segments of the kidney and other tissues negative for c-Jun and c-Fos and both c-Jun and c-Fos were present in the renal proximal and distal convoluted tubules, where GST-P was lacking. Thus, the localization of GST-P was in some cases clearly separable from those of c-Jun or c-Fos. GST-P was found to be focally expressed from an early stage of hepatocarcinogenesis, when c-Jun was not detectable. At later stages, this oncogene product was stained in 35.7% of GST-P-positive foci, with a clear relation to the degree of GST-P staining. Since GST-P is not always accompanied by appreciable c-Jun or c-Fos, these oncogene products are apparently not prerequisites for its expression. However, c-Jun may be partly responsible for maintaining high levels of GST-P in hepatic foci at later stages of hepatocarcinogenesis.

Animals

Decreased expression of glutathione S-transferases and increased fatty change in peroxisomal enzyme-negative foci induced by clofibrate in rat livers.

Alteration in glutathione S-transferase (GST) isoenzymes was compared with that of a peroxisomal enzyme, enoyl-CoA hydratase (ECH), during hepatocarcinogenesis caused by clofibrate (CF) administration in male Sprague-Dawley rats. The amount of alpha class GST forms, determined by single radial immunodiffusion using anti-GST 1-2 antibody, was inversely correlated with that of ECH and was decreased at week 2 of CF administration to approximately 50% of the value prior to treatment and then slightly increased to 70% of the control value by week 15, without change thereafter up to 93 weeks. Resolution of GST subunits by high performance liquid chromatography revealed an approximately 60% decrease in the amounts of subunits 1 and 3 at week 93 and a 25% decrease in the amounts of subunits 2 and 4. Immunohistochemical staining of rat livers revealed hepatic foci and minifoci negative for ECH at week 60 and thereafter. Almost all ECH-negative foci (95.1-97.9%) were clear cell in character, along with a somewhat lower proportion (68.0-73.8%) of ECH-negative minifoci. Numerous fat-positive granules were detected in 78.3% of those lesions exhibiting a clear cell change. Although the amounts of GST and ECH exhibited contrasting patterns of alteration in whole livers following CF administration, the expression of both alpha and mu class GST forms was decreased in the majority of ECH-negative foci at week 93, but were not altered in minifoci. The repression of GST forms appeared to be a later event than the loss of ECH or the clear cell change in CF-associated hepatic lesions and was in clear contrast to the enhanced expression reported for preneoplastic lesions induced by mutagenic carcinogens.

Animals

Left renal pelvis of male neonates is predisposed to dilatation.

Dilatation of the renal pelvis has been observed as an ultrasonographic finding of ureteral reflux as well as hydronephrosis. However, little information is available on the prevalence of renal pelvis at dilatation in neonates. We measured the inner pelvis dimension of the kidneys in 511 apparently healthy neonates (279 boys and 232 girls) using an ultrasound scanner to determine the prevalence of renal pelvis dilatation. Ninety per cent of the neonates had an inner dimension of both renal pelvises below 5 mm. The prevalence of left renal pelvis dilatation of 5 mm or more was significantly higher in the boys than in the girls, 25 (9%) compared to 5 (2%). In contrast, no significant difference was found in the prevalence of right renal pelvis dilatation between the sexes. In the boys, the prevalence of renal pelvis dilatation of 6 mm or more was significantly higher on the left side than on the right. Moreover, the left renal pelvis dilatation of the male neonates had a tendency to persist at 1 month of age. These findings suggest that the left renal pelvis of the baby boy may be predisposed to dilatation.

Chi-Square Distribution

Urine microscopy on a counting chamber for diagnosis of urinary infection.

Several quantitative methods of urine microscopic examination for bacteriuria and pyuria on a blood cell counting-chamber have been found reliable for the diagnosis of urinary tract infection (UTI). However, no one technique has become popular or widely used because of laborious procedures associated with the method. We investigated the usefulness of microscopic examination of uncentrifuged urine on disposable counting-chambers. A total of 89 urine samples were obtained from 53 children (24 male and 29 female). Urine samples were examined for bacteriuria and pyuria using a disposable counting chamber and its reliability was analyzed in predicting significant bacteriuria defined by routine urine culture. Significant bacteriuria was diagnosed in 23 of 89 urine samples by urine culture. Microscopic urine examination on disposable counting-chambers was very easy without the need to set up or wash chambers and provided immediate information. Urine bacterial concentration determined by the counting-chamber method was closely correlated to that determined by bacterial culture. The counting-chamber method identified bacteriuria correctly in 21 of 23 urine samples diagnosed as significant bacteriuria (sensitivity = 91%) and also gave a correct diagnosis of 64 of 66 urine samples with non-significant bacteriuria (specificity = 98%). Nineteen of the 23 urine samples with significant bacteriuria also had pyuria. The positive predictive value of concomitant bacteriuria and pyuria was 100%. When neither bacteriuria nor pyuria was found, the negative predictive value was 100%. It was concluded that urine microscopy using disposable counting chambers was very easy, inexpensive, quick and reliable and thus an extremely useful method for diagnosing UTI.

Adolescent

Hypercalciuria and nephrocalcinosis in patients with idiopathic low-molecular-weight proteinuria in Japan: is the disease identical to Dent's disease in United Kingdom?

Idiopathic low-molecular-weight (LMW) proteinuria is a newly described renal disease in Japan and Italy. We report on 7 patients who manifested bilateral or unilateral nephrocalcinosis, as demonstrated by abdominal computed tomography scans. Renal histology revealed calcinosis of renal tubules in 2 patients. Computed tomography is a reliable method for the detection of nephrocalcinosis in this disorder. Hypercalciuria was also seen in 6 patients. A calcium-loading test performed in 2 patients suggested that hypercalciuria was of renal origin. Although the true pathogenesis is still not known, hypercalciuria and nephrocalcinosis appear to be a common complication in patients with idiopathic LMW proteinuria. These complications and clinical features suggest that idiopathic LMW proteinuria in Japan is likely to be identical to Dent's disease in the United Kingdom.

Adolescent

[A case of spontaneous mesenteric fibromatosis occurred from the mesentery of the small intestine].

The fibromatosis is benign tumor which is characterized by the remarkable proliferation and the invasive growth of fibrous tissue and no distant metastasis. It usually occurs from the abdominal wall or the extremities, and rarely from the mesentery. A 54-year-old male complained of an epigastralgia and emesis. He was diagnosed through imaging as the obstructive ileus by abdominal tumor. The laparotomy was performed and a round, egg sized tumor was found wear by dilated ileum. The tumor was considered to occur from the mesentery, and the iliectomy with the tumor was completed. It was suggested that ileus occurred in the early stage because the tumor was located just by the ileum.

Fibromatosis, Abdominal

Molecular analysis of esterase D polymorphism.

We have analyzed the esterase D (EsD) polymorphism at the nucleic acid level. Two common alleles, EsD1 and EsD2, are characterized by the substitution of one amino acid (Gly-to-Glu), which is caused by the point mutation of one nucleotide (G-to-A). Individuals exhibiting the EsD1 and EsD 2 phenotypes are homozygotes for EsD 1 and EsD 2 cDNAs, respectively. Individuals showing the EsD 2-1 phenotype have two kinds of cDNAs, viz., EsD 1 and EsD 2. The point mutation difference between the cDNAs of the EsD1 and EsD2 alleles results in a different SspI digestion site. A restriction fragment length polymorphism caused by this difference with respect to the SspI digestion site makes it possible to determine the EsD phenotype using DNA samples extracted from forensic materials with no EsD enzymatic activity.

Amino Acid Sequence

Assessment of the pathological grade of astrocytic gliomas using an MRI score.

To evaluate the usefulness of an MRI score for identifying tumour tissue characteristics, 41 histologically verified supratentorial astrocytic gliomas, including 13 low-grade astrocytomas (LGA) 14 anaplastic astrocytomas (AA) and 14 glioblastoma multiformes (GBM), were examined with a 0.5T superconductive MR imager. Nine MRI criteria were used: heterogeneity (HET), cyst formation or necrosis (CN), haemorrhage (HEM), crossing the midline (CM), oedema or mass effect (EM), border definition (BD), flow void (FV), degree (CE-D) and heterogeneity (CE-HET) of contrast enhancement; Gd-enhanced T1-weighted images were obtained in 32 cases (10 LGA, 10 AA, and 12 GBM). Each of the criteria was scored and analysed statistically. The mean values of LGA, AA and GBM were 0.45 +/- 0.31, 1.18 +/- 0.20, and 1.47 +/- 0.22, respectively. The MRI score increased with the pathological grades (P < 0.01-0.001). LGA had significantly lower values than AA in five (HET, CN, EM, BD, CE-D) of the nine criteria (55.6%) and lower values than GBM in all except HEM (88.9%). Three criteria (33.3%): HET, CN, and FV were significantly higher in GBM than AA. CE-D, HET, EM, CN, and CE-HET proved to be related to the pathological grade by a multiple regression analysis (P < 0.001).

Adolescent