Biomedical subjects

A Yoshida

Publications and source records attributed to A Yoshida.

At least 469 records · Page 26Linked to original sources

PubMed1995

Cloning and expression of a porcine UDP-GalNAc: polypeptide N-acetylgalactosaminyl transferase.

By employing a bovine UDP-N-acetylgalactosamine: polypeptide N-acetylgalactosaminyl transferase (O-GalNAc transferase) cDNA as a probe, we isolated four overlapping cDNAs from a porcine lung cDNA library. Both the nucleotide sequence of the porcine cDNA and the predicted primary structure of the protein (559 amino acids) proved to be very similar to those of the bovine enzyme (95% and 99% identity, respectively). Transient expression of the clone in COS-7 cells, followed by enzymatic activity assays, demonstrated that this cDNA sequence encodes a porcine O-GalNAc transferase. The intracellular O-GalNAc transferase activity was increased approximately 100-fold by transfecting cells with the porcine cDNA.

Amino Acid Sequence↗

PubMed1995

Type 3 GM1 gangliosidosis: clinical and neuroradiological findings in an 11-year-old girl.

An 11-year-old Japanese girl was diagnosed as having type 3 GM1 gangliosidosis by clinical symptoms and enzyme assay. She was the youngest among the patients with type 3 GM1 gangliosidosis whose clinical and neuroradiological findings have been documented. Clumsiness since early infancy and dystonia since early childhood which progressed slowly without mental deterioration and dysmorphism led us to the diagnosis of type 3 GM1 gangliosidosis. Genotype determination showed point mutation in exon 2 of the beta-galactosidase gene, which is common among the patients reported in Japan. T2-weighted MRI demonstrated bilateral symmetrical hypointensity in the putamen and globus pallidus. Single photon emission computed tomography using 99mTc-HMPAO showed bilateral hyperperfusion in the basal ganglia which decreased gradually during 1 year of observation. Twenty-two patients with type 3 GM1 gangliosidosis reported in the literature whose onset was at under 15 years of age were reviewed.

Child↗

PubMed1995

Elevation of serum creatine phosphokinase during growth hormone treatment in patients with multiple pituitary hormone deficiency.

Serum creatinine phosphokinase (s-CPK) increased to more than 500 U/l in 5 out of 21 patients with growth hormone (GH) deficiency during the 2 years of treatment with biosynthetic GH. In three of these five patients, s-CPK had elevated gradually after the start of GH treatment and remained high in one patient except in the period when GH injection was interrupted, and gradually decreased in the other two patients during treatment. These three patients had complete GH deficiency associated with multiple pituitary hormone deficiency due to pituitary stalk transection. One of the remaining two patients had Noonan syndrome and his s-CPK levels before therapy were relatively high. The fifth patient was a baseball athlete and the elevation of s-CPK seemed to be attributable to the strenuous exercise. Conclusion. s-CPK increases significantly in a certain group of patients with GH deficiency during GH replacement therapy. Measurement of s-CPK is to be included in the follow up laboratory tests at least in the 1st treatment year to evaluate the potential hazardous effects of GH on muscle.

Adolescent↗

PubMed1995

The bovine rod outer segment guanylate cyclase, ROS-GC, is present in both outer segment and synaptic layers of the retina.

Cyclic-GMP, which plays a pivotal role in visual transduction in the vertebrate retina, is synthesized by guanylate cyclase. The purpose of this study was to localize a rod outer segment-derived particulate guanylate cyclase (ROS-GC) to the retina of several species that have different populations of rods and cones. A rabbit antibody was raised against a synthetic peptide, corresponding to the sequence A107-L125 of bovine ROS-GC. Western blot analysis showed a single immunoreactive band at about 115 kDa with bovine rod outer segments but not with human rod outer segments. Light microscopic immunocytochemistry of tissue sections revealed immunoreactivity in the outer segment layer and in the outer and inner plexiform layers. The rod-rich rat retina showed uniform immunolabeling of outer segments; the cone-containing cat retina showed heavily labeled cone outer segments and lighter labeling of rod outer segments; the cone-rich chicken retina showed a uniformly and intensely labeled outer segment layer. Preincubation of the primary antibody with the peptide completely blocked antibody binding. Electron microscopic immunocytochemistry of the cat retina confirmed the presence of guanylate cyclase in photoreceptor outer segments and demonstrated its association with disk and plasma membranes. These data support a concept in which guanylate cyclase is much more concentrated in the outer segments of cones than rods. The immunolabeling of the plexiform layers suggests that the particulate guanylate cyclase is not unique to the photoreceptor outer segments, and may also play a role in transduction processes of retinal synapses.

Amino Acid Sequence↗

PubMed1995

Dual-energy subtraction mammography.

Dual-energy subtraction mammography was performed for breast examinations. To obtain a dual-energy subtraction image with a digital radiography unit, high- and low-energy images were obtained at an appropriate time interval under different x-ray exposure conditions. In about 50% of the patients with breast cancer included in this study, we obtained better diagnostic accuracy with dual-energy subtraction images than with conventional mammography. In some cases of breast cancer, it is possible to diagnose intraductal spread of this lesion on the subtracted images. Furthermore, abnormal lesions commonly observed on mammography in cases of fibrocystic disease tended to be erased on subtracted images. Thus, dual-energy subtraction mammography provided useful information for diagnosing breast diseases. However, there were several cases in which the subtracted images lacked sufficient image quality, and several technical problems with subtraction are thought to remain.

Breast Neoplasms↗

PubMed1995

Application of in situ hybridization with a novel phenytoin-labeled probe to conventional formalin-fixed, paraffin-embedded tissue sections.

Non-isotopic in situ hybridization with a novel phenytoin (PHE)-labeled probe was developed. The mixture of cloned cytomegalovirus (CMV) DNA fragments was labeled by random primer technique using PHE-11(spacer)-dUTP, instead of dTTP. The tissue sections were treated with 0.2 N HCl and with proteinase K (1 microgram/ml), and then heated at 70 degrees C in the presence of 50 or 75% formamide. The sections were hybridized with PHE-labeled probe at 37 degrees C overnight. The hybridization signal was visualized by alkaline phosphatase-5-bromo-4-chloro-3-indolyl phosphate (BCIP)/4-nitroblue tetazolium (NBT) system. Strong hybridization signals were detected in sections of the small intestine and the placenta, even when denatured in the presence of 50% formamide. In the case of small intestine, CMV DNA was also detected in the endothelial cells of the mucosa where apparent infected cell was not observed histologically. In the sections of the submaxillary gland, the lung, the adrenal gland and the ovary, hybridization signal was not detected when denatured in the presence of 50% formamide, but detected after denaturation with 75% formamide. Thus, in situ hybridization with the novel PHE-labeled probe is applicable to conventional formalin-fixed, paraffin-embedded tissue sections.

Adrenal Glands↗

PubMed1995

Prognosis of stage 2 macular holes.

PURPOSE: We ascertained the natural course of stage 2 idiopathic macular holes to determine better treatment possibilities. METHODS: We reviewed 48 eyes with stage 2 idiopathic macular holes and followed them up for more than two years. At each examination, best-corrected Snellen visual acuity was measured by a physician masked to the hypothesis of the study. RESULTS: Stage 2 lesions progressed to stage 3 or 4 during the follow-up period in 32 (67%) and 14 (29%) of 48 eyes, respectively; two eyes (4%) remained in stage 2. In 41 (85%) of 48 eyes, the hole size enlarged during the follow-up: 32 (94%) of 34 eyes had vitreomacular attachment and nine (64%) of 14 eyes had vitreomacular separation at the final examination, for a statistically significant difference in prevalence (P = .03). Visual acuity decreased two or more Snellen lines during the follow-up period in 34 (71%) of 48 eyes, the prevalence of which was significantly higher in eyes with vitreomacular attachment at the final examination (28 of 34, 82%) than in eyes with vitreomacular separation at the final examination (six of 14, 43%) (P = .01). CONCLUSION: Even though vitreomacular separation may improve the prognosis of a macular hole, stage 2 lesions usually will develop an enlarged hole and decreased visual acuity.

Aged↗

PubMed1995

Course of vitreomacular traction syndrome.

PURPOSE: Although the course of vitreomacular traction syndrome is necessary to treat patients or to establish the value of the surgical technique, little information is available about its course. In this study, we ascertained the natural history of vitreomacular traction syndrome. METHODS: We retrospectively studied 53 consecutive symptomatic eyes with vitreomacular traction syndrome. RESULTS: In 43 (81%) of 53 eyes with cystoid macular changes at the diagnostic examination, 29 (67%) of 43 had cystoid changes that persisted during the median follow-up period of 60 months. The visual acuities at the time of the final examination decreased two Snellen lines or more from the initial measurement in 34 (64%) of 53 eyes. During the follow-up period, six (11%) of 53 eyes developed complete posterior vitreous detachment. The number of eyes with resolved cystoid changes or stable visual acuity was significantly higher when complete vitreomacular separation occurred (six of six) than when it did not (three [8%] of 37, P = .01; 13 [28%] of 47, P = .01). CONCLUSIONS: Most symptomatic eyes with vitreomacular traction syndrome underwent a further decrease in visual acuity. Complete vitreomacular separation, which occurs infrequently in eyes with the disorder, allows resolution of cystoid changes and improvement of visual acuity.

Aged↗

PubMed1995

Antigens associated with N- and L-type calcium channels in Lambert-Eaton myasthenic syndrome.

In Lambert-Eaton myasthenic syndrome neurotransmitter release is reduced by an autoimmune response directed against the calcium channel complex of the nerve terminal. Autoantibodies were detected by immunoprecipitation assays using solubilized receptors labeled with ligands selective for N-type (125I-omega conotoxin GVIA) and L-type ([3H]PN200-110) calcium channels. Sera with a high antibody titer (> 3 nM) against rat brain N-type channels contained autoantibodies that immunoprecipitated neuronal and muscle L-type channels. These IgG fractions stained a 55-kDa protein in immunoblots of purified skeletal muscle dihydropyridine receptor, suggesting that they contain autoantibodies against the beta subunit of the calcium channel. A distinct antibody population in the same fractions reacted with a nerve terminal 65-kDa protein that is unrelated to the beta subunit and displays properties similar to those of synaptotagmin.

Animals↗

PubMed1995

Cell swelling has differential effects on the rapid and slow components of delayed rectifier potassium current in guinea pig cardiac myocytes.

Cell swelling has been shown to cause activation of a variety of cardiac sarcolemmal ionic conductances including potassium channels. The aim of this study was to investigate the effect of swelling on the two subtypes of delayed rectifier potassium current (IKr and IKs) in single guinea pig myocytes using the whole-cell configuration of the patch clamp technique. When the holding potential was set at -40 mV and stepped to +40 mV for 1 s under isoosmotic conditions (300 mOsm) a delayed rectifier current (IK) was activated (0.86 +/- 0.05 nA; n = 43). Switching to a hypoosmotic solution (200 mOsm) caused a rapid increase in IK to a mean value of 1.43 +/- 0.10 nA (p < 0.05; n = 43). The effect of swelling on the two subtypes of IK was studied by analysis of deactivating tail currents using an envelope of tails protocol (stepping from -40 to +40 mV for 18 different pulse durations between 50 ms and 2.9 s; n = 16). Swelling caused a decrease in current amplitude measured at the end of the pulse (and IKtail) at short durations (< or = 150 ms) however, when the pulse duration was > 1 s swelling caused a significant increase in current. Using a pulse protocol to measure IKr with minimal contamination by IKs (voltage step from -40 to -10 mV for 250 ms) a 50-100 pA current was elicited which could be completely blocked by dofetilide (0.2 microM; n = 3). Introduction of hypoosmotic solution caused a significant decrease in IKr and when dofetilide (0.2 or 1.0 microM) was introduced the current remaining was decreased further (p < 0.05; n = 5), but was not completely blocked, thus suggesting that swelling had decreased the ability of dofetilide to block IKr. Similar results were obtained over a range of dofetilide concentrations and with a second IKr blocker, La3+. In Ca(2+)-free external solutions, pulsing to -10 mV for 500 ms to measure IKr in the absence of IKs, and to +60 mV for 5 s (with 0.2 microM dofetilide) to evoke only IKs, it was clear that swelling significantly increased IKs (pulse and tail currents) and decreased IKr. In addition, when measured using the perforated patch method, swelling modulated IKt and IKs in a similar fashion. We conclude that swelling has differential effects on the subtypes of the classical cardiac IK, which may have important implications in our understanding of the mechanisms underlying ischaemia- and reperfusion-induced arrhythmogenesis.

Animals↗

PubMed1995

A patient with an autonomously functioning thyroid nodule with papillary adenocarcinoma associated with Graves' hyperthyroidism.

A 69-year-old man had concomitant findings of an autonomously functioning thyroid papillary adenocarcinoma and Graves' hyperthyroidism. The patient presented with hyperthyroidism and a right lobe thyroid nodule that was shown to be hyperfunctional on I-123 imaging. The TI-201 imaging showed uptake confined almost entirely to the right lobe nodule. Papillary adenocarcinoma in the nodule and diffuse hyperplasia surrounding the nodule along with a slightly enlarged left lobe were found at surgery.

Adenocarcinoma, Papillary↗

PubMed1995

Vitreous observation using a CCD camera and a computerized unit for image processing and storage.

PURPOSE: This study was conducted to evaluate a new technique of vitreous observation. METHODS: The authors used commercially available devices, including a slit-lamp, monochromatic charge coupled device (CCD) camera, and a computerized unit for image processing and storage, to observe the vitreous and the vitreoretinal relationship. RESULTS: The authors obtained good images of the vitreous and the vitreoretinal relationship using the new technique. Furthermore, the image stored in the computer could be accessed and used for patient explanations instantly. CONCLUSION: This technique is useful for documenting the vitreous and can help determine the role of the vitreous in many vitreoretinal disorders.

Aged↗

PubMed1995

Enhancement of Ca(2+)-dependent endonuclease activity in L1210 cells during apoptosis induced by 1-beta-D-arabinofuranosylcytosine: possible involvement of activating factor(s).

Internucleosomal DNA fragmentation and morphological changes in nuclei typical of apoptosis were observed in L1210 cells incubated with 1.0 micrograms/ml of 1-beta-D-arabinofuranosylcytosine (ara-C). To investigate the mechanisms involved, we examined the activities of endogenous endonucleases in nuclei and cytoplasm. Both fractions of control cells contained Ca(2+)-dependent endonuclease which was capable of mediating internucleosomal DNA fragmentation. The assay system using two kinds of target substrates, i.e., nuclear chromatin of CCRF-CEM cells and naked DNA purified from the same cells, revealed that the activity of Ca(2+)-dependent endonuclease was enhanced in the crude nuclear extracts of cells treated with 1.0 microgram/ml of ara-C for 24 h or 48 h. The activity was extracted more easily from ara-C-treated cells than control cells without sonication of the nuclear fraction. On the other hand, in the cytoplasmic fraction of the cells, the activity towards naked DNA was unchanged, whereas that towards nuclear chromatin was clearly enhanced. These results suggest that internucleosomal DNA fragmentation induced by ara-C treatment is associated with enhancement and activation of constitutively expressed Ca(2+)-dependent endonuclease in L1210 cells.

Animals↗

PubMed1995

Molecular defect of a phosphoglycerate kinase variant associated with haemolytic anaemia and neurological disorders in a large kindred.

The X-chromosome-linked phosphoglycerate kinase (PGK) deficiency associated with severe chronic and acute haemolytic anaemia and mental disorders was first described in a large Chinese kindred in 1969. The molecular abnormality of this original variant remained to be identified. The red cell PGK activity was only about 5%, but the activity of the patients' lymphoblastoid cells was about 15% of normal. The PGK mRNA content of the patients' lymphoblastoid cells were normal. Analysis of the patients' mRNA showed the existence of a nucleotide transversion A-->T at position 491 (counting from adenine of the initiation codon). The mutation should cause an amino acid substitution Asp-->Val at position 163 of the enzyme. The replacement of the acidic aspartic acid by a hydrophobic valine is expected to induce drastic structural instability resulting in severe enzyme deficiency in the patients' tissues. The genotypes of two affected males, their mothers and 22 females of the family were identified by the PCR-mediated method using their genomic DNA samples. 13/24 females examined were found to be variant heterozygous. In this large family, affected males over three generations have died at a pre-adult age. Post- and pre-natal genotyping of the family members may prevent future problems.

Anemia, Hemolytic, Congenital↗

PubMed1995

Structure of the human deoxyribonuclease I (DNase I) gene: identification of the nucleotide substitution that generates its classical genetic polymorphism.

The objectives of this study were to elucidate the structural organization of the gene for human deoxyribonuclease I (DNase I) and to identify the mutation site underlying its classical genetic polymorphism. In order to determine the organization of this gene, we utilized a combination of direct polymerase chain reaction (PCR)-amplification of human genomic DNA and isolation of the overlapping clones from a cosmid human genomic library. Restriction endonuclease mapping, Southern blotting and DNA sequencing showed that the DNase I gene was approximately 3.2 kilobases long, it comprised 9 (I-IX) exons separated by eight introns and its complete sequence was determined. The first exon contained only the non-translated sequences of mRNA. In addition to several putative regulatory elements, TATA-like and CAAT-like sequences were observed in the region upstream of the translation initiation codon. These results provide information that will help to understand the expression and regulation of DNase I. The isoelectric focusing patterns of human DNase I showed that it exhibits classical genetic polymorphism (Kishi et al. 1989, 1990). A comparison of the entire translated sequences of the DNase I gene from two pairs of individuals with common DNase I phenotypes 1 and 2 revealed only one nucleotide residue difference in exon VIII, A for phenotype 1 and G for phenotype 2, thus producing Gln and Arg amino acid substitutions respectively at position 222 from the NH2-terminus of the mature enzyme. The predicted charge changes attributable to these amino acid substitutions are entirely consistent with the isoelectric focusing profiles of these two DNase I isozymes. We conclude that this substitution is solely responsible for the classical polymorphism of DNase I protein.

Adult↗

PubMed1995

A simple and effective video keratometric system.

We tested a personal computer-based video keratometric system used to analyze projected placido disk rings. The system consists of a Maloney surgical keratometer that projects the rings and a video camera attached to an operating microscope. Images of the rings were transmitted to a video image processing board and analyzed on a personal computer using an analysis program we developed. System precision was evaluated in 18 eyes. The values were compared with those obtained using a commercial photokeratometer (control). In all eyes, the principal meridian values differed from the controls by < 0.04 mm. In 15 eyes (83%), the measurements were within 0.03 mm of the controls. The corneal astigmatism values and cylindrical axes were within 0.12 diopters in 17 eyes (94%) and within 5 degrees in all cases, respectively. Our system, which can analyze placido disc rings projected from other systems, is sufficiently precise to measure the corneal radius of curvature and astigmatism.

Calibration↗

PubMed1995

Natural outcomes of stage 1, 2, 3, and 4 idiopathic macular holes.

AIMS: A study was carried out to ascertain the natural outcome of each stage of idiopathic macular hole. METHODS: One hundred and fifty four eyes with different stages of idiopathic macular holes were retrospectively studied: stage 1 (40 eyes), 2 (25 eyes), 3 (58 eyes), and 4 (31 eyes). RESULTS: Of 27 of 40 eyes with a stage 1 lesion with posterior vitreous attachment to the macula initially, nine (33%) eyes developed a full thickness macular hole. No stage 1 lesions with posterior vitreous separation from the macula initially progressed to full thickness holes. Twenty one (84%) of 25 eyes with a stage 2 lesion, 32 (55%) of 58 eyes with a stage 3 lesion, and five (16%) of 31 eyes with a stage 4 lesion underwent macular hole enlargement during the median follow up period of 3 years. Visual acuity decreased two or more lines of Snellen equivalent during the follow up period in 12 (30%) eyes with a stage 1 lesion, 17 (68%) eyes with a stage 2 lesion, 17 (29%) eyes with a stage 3 lesion, and four (13%) eyes with a stage 4 lesion. The percentage was significantly higher in eyes with stage 2 lesions than the other stages (p < 0.01). CONCLUSION: The results suggest that the different stages of idiopathic macular holes have different natural outcomes, and the management depends on the stage at presentation.

Age Distribution↗

PubMed1995

Changes in lens organophosphate metabolites in response to endotoxin-induced uveitis.

Uveitis was induced in rats by endotoxin administration. The temporal correlation between ocular inflammatory reaction and lens metabolic change was examined. The levels of lens phosphate compounds, adenosine triphosphate (ATP), choline phosphate (CP), and inorganic phosphate (Pi), were measured using 31P-nuclear magnetic resonance (NMR) spectroscopy. Anterior chamber inflammation reaction appeared 12 h after administration, peaked at 24 h, and disappeared by 72 h. Concurrent with the active inflammation, lens Pi increased significantly 22 +/- 17% (p < 0.05) and 23 +/- 15% (p < 0.05), 12 and 24 h after administration, respectively, and returned to normal while the inflammation abated. No significant changes were observed in CP and ATP levels in the lens during the course of inflammation. The time course of the inflammation appeared closely related to the change in Pi level, which may be part of the protective mechanism of the lens against inflammation.

Adenosine Triphosphate↗