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Y Hosoya

Publications and source records attributed to Y Hosoya.

At least 73 records · Page 4Linked to original sources
PubMed1996

Identification of 29 rat genetic markers by arbitrarily primed polymerase chain reaction.

The number of genetic markers for the rat is still limited, in spite of its wide use in cancer research. To facilitate accurate mapping of both established and novel rat genetic markers, we constructed a linkage map by genotyping 105 F2 rats from ACI/N (ACI) and BUF/Nac (BUF) crosses. This map consists of 120 genetic markers that had been previously reported, mainly by two research groups, but had not been integrated. To find new genetic markers, the arbitrarily primed polymerase chain reaction (AP-PCR) was applied to detect polymorphic bands between ACI and BUF rats. After testing 56 single primers and 12 combinations of primers, we found 36 bands produced by 16 single primers and two combinations to be reliably polymorphic between ACI and BUF rats. The 36 bands were typed in the 105 F2 rats, and 29 of them could be linkage-mapped. AP-PCR is thus useful to detect new genetic markers in laboratory strains of rats.

Animals↗
PubMed1996

[Determination of free type and complex type prostate-specific antigen (PSA): differences in immunorecognition by Delfia PSA, ACS-PSA and Eiken PA kits].

To study immunorecognition of free type and complex type prostate-specific antigen (PSA) by current commercial PSA assays, sera from 3 patients with stage D2 prostate cancer were separated by Sephacryl S-200 chromatography and determined by Delfia PSA, ACS-PSA and Eiken PA kits. Two antibodies used in the 3 kits are 2 monoclonal, 1 monoclonal and 1 polyclonal and 2 polyclonal antibodies, respectively. Following chromatography, two PSA peaks were obtained in all patients. One was about 100 kDa and the other about 30 kDa. The former was considered to be the complex type PSA (complex with alpha-1 antichymotrypsin) and the latter to be free type PSA. As to free type PSA, the ACS-PSA kit and Eiken PA kit quantitated PSA values approximately 5.1 and 2.5 times higher than the Delfia PSA kit. For complex type PSA, the quantity determined by ACS-PSA kit was approximately 1.3 times higher than that determined by the Delfia PSA kit, while the quantity determined by the Eiken PA kit was about one third of that determined by Delfia PSA kit. The ratio of complex type PSA to total PSA (free type PSA + complex type PSA) was 74.8 +/- 14.9% (mean +/- SD) when determined by Delfia PSA kit, 59.3 +/- 18.4% by ACS-PSA kit and 52.9 +/- 13.8% by Eiken PA kit. The range of this ratio determined by ACS-PSA kit was from 47.3% to 80.5% in the 3 patients. These findings suggest that there are qualitative differences in immunorecognition of free type PSA and complex type PSA among current commercial PSA assays and that there are quantitative differences in the ratio of the 2 forms of PSA in serum among prostatic cancer patients. The measurement and follow-up of both free type and complex type PSA might be important for diagnosis and monitoring of prostate cancer.

Aged↗
PubMed1996

[The significance of free-type PSA and complex-type PSA in patients with prostatic carcinoma--the characteristics of ACS-PSA method compared with that of Delfia- and Eiken-PSA method].

We studied the clinical significance of free-type prostate specific antigen (PSA) and bound-type PSA in serum of the patients with prostatic carcinoma and benign prostatic hypertrophy. The levels of PSA, gamma-seminoprotein and prostatic acid phosphatase (PAP) in 17 healthy adult males, 20 patients with benign prostatic hypertrophy and 23 patients with prostatic carcinoma were measured by ACS-PSA, Delfia and Eiken-PSA method. The levels of PSA in serum from prostatic carcinoma patients was significantly elevated as compared with that from benign prostatic hypertrophy. Linear regression analysis of the data showed that, although overall correlations were well, different assays gave different PSA concentrations. We have studied the forms of PSA in serum by gel filtration technique and measured PSA levels in each fractions using three methods. Moreover, the characteristics of ACS-PSA method was compared with that of Delfia and Eiken-PSA method. Two peaks of PSA were detected on the elution profiles from three prostatic carcinoma patients sera. Those were estimated complex-type PSA (90kDa) and free-type PSA (30kDa). The complex-type PSA fractions detected by ACS-PSA method were almost identical with that detected by Delfia-PSA method, while free-type PSA fractions detected by ACS-PSA method were greater quantity than those by Delfia and Eiken method. Many factors were contributory to the difference between the assay kits on serum complex-type PSA and free-type PSA levels. The present results suggest that there are some quantitative differences in the immunorecognition of complex-type PSA and free-type PSA between the assay kits.

Adult↗
PubMed1996

[Studies on changes in the ratio of free to total PSA after endocrine treatment of prostate carcinoma].

The post-diagnostic changes in the free to total PSA ratio in the serum of patients with prostate carcinoma, after the initiation of endocrine treatment were examined. Two-week pretreatment with either chlormadinone acetate (100 mg/day) or flutamide (375 ng/day) was administered orally to 14 patients with newly diagnosed advanced prostate carcinoma (clinical stage was C in 2, Dl in 1 and D2 in 11). Then the LH-RH analogue was injected. Total and free PSA in the serum of these patients were measured every 4 weeks by the Ab bead PSA (Eiken) and the recently developed assay for free PSA by Eiken, respectively. The follow-up period ranged from 3 to 9 months with a median of 6 months. Levels of both total and free PSAs decreased significantly following the endocrine treatment, while free to total PSA ratio at 4 to 16 weeks after the start of LH-RH analogue was increased significantly compared to the pretreatment level (p < 0.05). These findings suggest that the rate of decrease of complex PSA during the first 4 months after the beginning of treatment may exceed that of free PSA in the serum of patients with advanced prostate carcinoma initially treated with endocrine therapy.

Aged↗
PubMed1995

Immunological and physical characterization of the brain G protein-gated muscarinic potassium channel.

The subunit composition of the G protein-gated inwardly rectifying K+ (KG) channel protein extracted from mouse forebrain membranes was examined. A polyclonal antibody (anti-GIRK1C1) was prepared against the carboxyl terminal region of GIRK1, the major subunit of the KG channel. The anti-GIRK1C1 IgG detected a single protein at approximately 65 kDa in SDS-PAGe of brain membranes. This IgG immunoprecipitated a macroprotein complex composed of GIRK1 and several associated proteins whose molecular weights ranging from 50 to 62 kDa on SDS-polyacrylamide gel. Upon size fractionation by both sucrose density gradient centrifugation and gel filtration, the solubilized KG channel proteins migrated into a single peak, which suggests that the component subunits form a single macromolecule. On the basis of the values of size fractionation, the molecular mass of the KG channel macromolecule could be estimated at approximately 231,000. These results suggest that the KG channel is most likely a tetrameric macroprotein composed of GIRK1 and co-immunoprecipitated proteins in the forebrain.

Amino Acid Sequence↗
PubMed1995

Serial lectin affinity chromatography demonstrates altered asparagine-linked sugar chain structures of gamma-glutamyltransferase in human renal cell carcinoma.

Differences between human renal cortex and human renal cell carcinoma (RCC) in asparagine (Asn)-linked sugar chain structures of gamma-glutamyltransferase (GGT) were investigated by using a serial lectin affinity chromatographic technique. The relative amounts of GGT which passed through the concanavalin A (Con A) column but bound to the phytohaemagglutinin E column, were significantly decreased in RCC, but there were significant increases in the relative amounts of GGT which bound weakly to the Con A column and passed through the pea lectin (PSA) column, and bound strongly to the Con A column and bound to the wheat germ agglutinin column in RCC compared with those of the normal renal cortex. A significant correlation was observed in RCC between nuclear grade and relative amount of GGT which bound weakly to the Con A column and passed through the PSA column. The findings indicate that Asn-linked sugar chain structures are altered in RCC and suggest that studies of qualitative differences of sugar chain structures of GGT might lead to a useful diagnostic tool for human RCC.

Adult↗
PubMed1995

Mutation, loss of heterozygosity, and recombination of the p53 gene in mouse forestomach tumors induced by 2-amino-3,4-dimethylimidazo[4,5-f]quinoline.

2-Amino-3,4-dimethylimidazo[4,5-f]quinoline (MeIQ), a food mutagen, induces forestomach tumors in CDF1 mice. We established a polymerase chain reaction (PCR)-single-strand conformation polymorphism (SSCP) analysis system to detect mutations in the mouse p53 gene exons 2-10, which encompass all five regions conserved among species, and a system to examine loss of heterozygosity (LOH) that uses newly identified polymorphisms between BALB/c and DBA mice, the parental strains of CDF1 mice. Four original forestomach tumors (one papilloma, two carcinomas, and one lymph-node metastasis) and four cell lines derived from four independent forestomach tumors were examined with the PCR-SSCP system and by polymorphism analysis. Of the four original tumors, the papilloma had a G-->A transition at the second position of codon 171, and one carcinoma had a G-->T transversion at the second position of codon 113 with loss of the wild-type allele, whereas the other two carcinomas had no detectable mutations. Of the four cell lines, two had a base substitution and LOH, and the other two had double mutations (a base substitution and a deletion). By amplification of the double mutations in a fragment, the two cell lines were shown to have four kinds of alleles, indicating induction of recombination within the p53 gene. Our results show that our PCR-SSCP analysis system is efficient for detecting p53 mutations in mouse genomic DNA and that alteration of the p53 gene plays a significant role in MeIQ-induced mouse forestomach carcinogenesis.

Animals↗
PubMed1995

Acetylcholine and adenosine activate the G protein-gated muscarinic K+ channel in ferret ventricular myocytes.

The properties of the K+ channel activated by acetylcholine (ACh) and adenosine (Ado) were examined in single ferret ventricular myocytes using patch-clamp techniques. In the whole-cell configuration, ACh and Ado induced an inwardly rectifying K+ current and shortened the action potential duration. The effect of ACh was blocked by atropine, while the Ado effect was interrupted by 8-cyclopentyl,1,2-dipropyl xanthine, a specific Ado A1 receptor antagonist. In cell-attached recordings, ACh and Ado added to the pipette solution activated a single population of inwardly rectifying K+ channels, distinct from the iK1 channel. The channel had a slope conductance of approximately 40 pS in symmetrical 150 mM K+ solutions and a mean open time of 0.8 ms. Excision of the patch into the inside-out patch configuration in guanosine triphosphate (GTP)-free solution abolished the channel activity. The channel was reversibly reactivated by adding GTP to the intracellular side of the patch. GTP gamma S activated the channel irreversibly. When the inside-out patch was treated with the A protomer of pertussis toxin (PTX), intracellular GTP no longer activated the K+ channel. The results show that ferret ventricular myocytes possess a K+ channel activated by both muscarinic and Ado A1 receptors. Its electrophysiological properties and the gating by a PTX-sensitive G protein in a membrane-delimited fashion are identical with those of the muscarinic K+ channels in nodal and atrial tissues of other species. In conclusion, the G protein-gated muscarinic K+ channel is expressed in ferret ventricular myocardium and may underlie the direct negative inotropism of ACh and Ado in this tissue.

Acetylcholine↗
PubMed1995

Oxytocinergic innervation to the upper thoracic sympathetic preganglionic neurons in the rat. A light and electron microscopical study using a combined retrograde transport and immunocytochemical technique.

A combination of retrograde cell body labeling and immunohistochemistry was employed to elucidate how oxytocinergic fibers make contact with sympathetic preganglionic neurons (SPNs) in the rat spinal cord from T1 to T4. SPNs were labeled retrogradely using cholera toxin subunit B (CTb) or horseradish peroxidase-conjugated CTb. Oxytocin-immunoreactive (ir) fibers were found in the intermediate zone, including the sympathetic preganglionic subnuclei. In the central autonomic nucleus and the intercalated nucleus, brown-stained oxytocin-ir varicosities or terminals were frequently observed to stud black-stained dendrites of SPNs. Electron microscopical observations showed that oxytocin-ir terminals form synapses with dendrites or soma of the sympathetic preganglionic neurons. The terminals contained numerous small clear round vesicles and a few large, cored vesicles. These results clearly show that a large proportion of SPNs are innervated by oxytocin-containing fibers. The origin of these fibers is discussed, and it is concluded that they are probably descending fibers from the paraventricular nucleus of the hypothalamus.

Animals↗
PubMed1995

A rapid method for detection of mutations in the lacI gene using PCR-single strand conformation polymorphism analysis: demonstration of its high sensitivity.

The lacI gene has been used as a target gene in various mutation assays. We modified single strand conformation polymorphism (SSCP) analysis by introducing restriction digestion to detect mutations in the gene rapidly, and determined the sensitivity of the method. The entire coding sequence and partial promoter region of the lacI gene were amplified by the polymerase chain reaction with [alpha-32P]dCTP in a 1247 base pair fragment, digested into eight restriction fragments, and analyzed by SSCP. The sensitivity of the method was assessed using 160 phages with lacI mutations, which were selected by assay of expression of beta-galactosidase after their infection into E. coli. Of the 160 mutants, 146 (91.3%) showed shifted bands in the first condition of SSCP analysis (without glycerol, 20 degrees C). The remaining 14 mutants were analyzed in a second condition (with 5% glycerol, 20 degrees C), and eight of them showed shifted bands (cumulatively 96.3% of the 160 mutants). The remaining six mutants were analyzed in a third condition (with 5% glycerol, 10 degrees C), and all of them showed shifted bands (cumulatively 100%). Sequencing of the restriction fragments with mobility shifts in the 160 mutants revealed 108 kinds of mutations, 100 (92.6%) being detected in the first condition, seven (cumulatively 99.1%) in the second condition, and one (cumulatively 100%) in the third condition. This method greatly reduced the time to identify lacI mutations, and allowed the detection of multiple mutations in one lacI mutant. The results also show that in general PCR-SSCP analysis is very sensitive when test fragments are shorter than about 250 base pairs and electrophoresis is performed under at least two conditions.

Animals↗
PubMed1995

Chemical compositions of 300 lower urinary tract calculi and associated disorders in the urinary tract.

The stones removed from 273 patients (220 males, 53 females) with bladder stones and 27 patients with urethral stones (males) were analyzed by infrared spectroscopy. Of these stones 50.0% contained magnesium ammonium phosphate (MAP), 39.9% calcium (oxalate and/or phosphate), 9.4% uric acid (urate), and 0.7% cystine. The disorders associated with bladder stones were prostatic hyperplasia and cancer (47.8%), neurogenic bladder (11.8%), urethral stricture (3.6%) and so on in males, and neurogenic bladder (48.1%), after operation of uterus (29.0%), bladder cancer (5.7%) and so on in females. Urinary infections were complications in 59.3% of patients with MAP stones, 41.2% with urate stones and 20.0% with calcium stones. The most often isolated bacteria from the infected urine were of the Proteus species.

Adolescent↗
PubMed1995

[Clinical evaluation of chemohormonal therapy as an initial treatment for stage D2 prostatic cancer--effect of UFT administration combined with hormonal therapy].

We evaluated the usefulness of hormonal therapy combined with UFT as initial treatment in comparison with hormonal therapy alone in 92 patients with Stage D2 prostatic cancer treated at the Department of Urology, Dokkyo University School of Medicine between 1974 and 1993. Twenty-six of these patients were treated with diethylstilbestrol diphosphate (DESP) and castration (hormonal therapy alone group), and 23 patients were treated with UFT, DESP and castration (UFT combined therapy group). The 5-year survival rates calculated with Kaplan-Meier's method in the hormonal therapy alone group and the UFT combined therapy group were 34.6 % and 38.3%, respectively. However, the 3-year survival rates of pathologically poorly differentiated adenocarcinoma in these groups were 30.0% and 50.0%, respectively. Based on these results, it was suggested that UFT administration combined with hormonal therapy is useful for pathologically poorly differentiated adenocarcinoma in Stage D2 prostatic cancer.

Adenocarcinoma↗
PubMed1995

[A case of lupus cystitis successfully treated with corticosteroid accompanied by gastrointestinal symptoms].

A 34-years-old woman was admitted to our department in February, 1992, because of nausea, vomiting, abdominal pain and diarrhea. She had been diagnosed as systemic lupus erythematosus (SLE) in 1988 and treated with prednisolone at the dose of 5 mg a day. In December, 1991, gastrointestinal symptoms developed followed by anuria on March 3, 1992. The laboratory findings revealed no activities in SLE. Computed tomography (CT) showed bilateral hydroureteronephrosis, swelling of bladder and gastrointestinal wall, and ascites. Under the diagnosis of lupus cystitis, corticosteroid therapy was started with 125 mg of methylprednisolone. Her symptoms improved immediately. Abnormal findings shown in the previous CT disappeared concomitantly. Lupus cystitis was reported by Orth et al. 1983 as severe fetal syndrome. However, because early corticosteroid therapy appears to reverse acute manifestation of lupus cystitis without complications, attention should be paid on lupus cystitis in patients with SLE with gastrointestinal symptoms of unknown etiology and decreasing urinary volume.

Adult↗
PubMed1995

Resin adhesion to the primary enamel: influence of light-irradiation times.

The purpose of this study was to investigate the influence of the light-irradiation times to the bonding agent and the composite resin on resin adhesion to the primary enamel. One hundred bovine mandibular ground primary incisors were used. Materials used in this study were 40% phosphoric acid gel, Photo Bond and Photo Clearfil A (Kuraray Co., Kurashiki, Japan). The etching time was 30 seconds. The light-irradiation times for the bonding agent were 10, 20 and 30 seconds and the light-irradiation times for the composite resin were 20, 40 and 60 seconds. Specimens were divided into the non-thermal cycled groups and the thermal cycled groups. Shear bond strengths were tested. After the shear bond strength test. the test surfaces of the enamel and the resin specimens were observed using the SEM. The optimum light-irradiation time on the bonding agent was 10 seconds. Both in the non-thermal cycled group and the thermal cycled group, the light irradiation time on the composite resin which showed the significantly higher bond strength was 40 seconds. Higher bond strength was not always obtained with the longer light-irradiation time on the bonding agent and the composite resin.

Acid Etching, Dental↗
PubMed1994

Thoracolumbar sympathetic preganglionic neurons in the dorsal commissural nucleus of the male rat: an immunohistochemical study using retrograde labeling of cholera toxin subunit B.

The cell morphology of sympathetic preganglionic neurons (SPNs) in the dorsal commissural nucleus was studied by the retrograde labeling technique using cholera toxin subunit B (CTb) as a tracer. A small amount of an aqueous solution of CTb was injected unilaterally into the major pelvic ganglion of the male rat. Labeled SPNs were detected immunohistochemically using anti-CTb antiserum. Most of the labeled SPNs were observed in L1 to L3, and a very small number in T13. They were observed bilaterally in the sympathetic nuclei, such as the intermediolateral cell column, intercalated nucleus and the dorsal commissural nucleus. A loose network of longitudinally or transversely oriented SPN dendrites was located within the dorsal commissural nucleus itself. The lateral margin of the dorsal commissural nucleus was roughly demarcated by longitudinally oriented dendrites. Together with the dendrites of the SPNs of the intercalated and intermediolateral cell column, laterally oriented dendrites of the dorsal commissural nucleus converged and formed the transverse dendritic bundles in the intermediate zone that connect the dorsal commissural nucleus and the intermediolateral cell column. The transverse dendritic bundles were arranged periodically. The axons of the SPNs in the dorsal commissural nucleus traveled laterally into the transverse dendritic bundles, then turned ventrally near the intermediolateral cell column, and finally entered the ventral funiculus. After rhizotomy of the ventral roots of the upper lumbar cord, labeled SPNs were found only on the side contralateral to the rhizotomy. The dorsal commissural nucleus appears as a compact single cell column, but our results clearly show that this nucleus actually consists of two adjacent parallel columns of cells.

Animals↗
PubMed1994

Dualistic behavior of ATP-sensitive K+ channels toward intracellular nucleoside diphosphates.

ATP-sensitive K+ (KATP) channels are intracellular ligand-gated channels which regulate diverse cellular functions. Intracellular nucleoside diphosphates (NDPs) are essential for the physiological opening of KATP channels which would otherwise be permanently closed by their overt sensitivity to intracellular ATP. We find that KATP channels exhibit dualistic behavior toward NDPs depending on their operative condition. When channels are in the spontaneous operative condition, NDPs antagonize channel inhibition by intracellular ATP. When channels have "run down", NDPs induce channel opening but no longer antagonize intracellular ATP. The switch of the KATP channel response to the same ligand, i.e., NDPs, is controlled by a Mg-ATP-dependent reaction. The condition of the target protein therefore determines the effect of the ligand. This property provides a novel basis to evaluate the dynamic regulation of ion channels by their ligands.

Adenosine Triphosphate↗