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Biomedical subjects

H Yoshioka

Publications and source records attributed to H Yoshioka.

At least 19 recordsLinked to original sources

Truncated type II receptor for BMP-4 induces secondary axial structures in Xenopus embryos.

BRK-3 is a vertebrate type II receptor for BMP-4 distantly related to invertebrate type II receptors for BMP-2/BMP-4/dpp, such as daf-4 and punt. BRK-3 has a long carboxy-terminal sequence following intracellular kinase domain and is capable of forming a high-affinity complex with a type I receptor, BRK-2. To examine the role of BRK-2 + BRK-3 receptor complex in BMP signaling during early embryogenesis, the dominant-negative form of BRK-3 was ectopically expressed in the Xenopus embryos. A secondary body axis expressing the Sonic hedgehog and N-CAM genes is induced by injecting mRNA encoding truncated form of BRK-3 into ventral marginal region, implicating the BMP signaling in axial mesoderm induction. Formation of the secondary axis depends on whether the deletion extends into the kinase domain, not into the carboxy-terminal tail, suggesting that the kinase domain, but not the tail region, is essential for BMP signaling.

Amino Acid Sequence

A chicken Wnt gene, Wnt-11, is involved in dermal development.

We have isolated a new member of the Wnt gene family, Wnt-11, from chick embryo cDNA library and examined the expression pattern during embryogenesis by in situ hybridization. The Wnt-11 gene encodes Cys-rich secretory protein distantly related to Wnt-1 through Wnt-8 from the mouse and Xenopus. Expression of the Wnt-11 gene became evident at stage 14 in the dorsolateral region of somites and gradually restricted to the dermatome at stage 19 and later. In contrast to the other Wnt genes, Wnt-11 was not expressed in the neuroepithelium throughout stages 14-26. At stage 24 and later, Wnt-11 was expressed in the subectodermal mesenchyme of the limb and feather buds. The unique expression pattern of Wnt-11 in the paraxial mesoderm and dermatome suggests that Wnt-11 may play an important role in dermal development.

Amino Acid Sequence

An additional limb can be induced from the flank of the chick embryo by FGF4.

To elucidate what initiates formation of the limb, we have attempted to induce an additional limb from the flank of the chick embryo by infecting retrovirus or implanting cells. We report here that an additional limb can be formed from the flank when we implant fibroblast growth factor 4 (Fgf4)-expressing cells into the lateral plate mesoderm at the pre-limb bud stage. In a newly formed limb bud, expressions of both Sonic hedgehog and chick Fgf4, which are authentic morphogenetic signals from the zone of polarizing activity and the apical ectodermal ridge, respectively, are induced by the implanted cells. Thus, it is concluded that the competence for limb development is present along the flank of the chick embryo and that FGF4 applied ectopically at the pre-limb bud stage can alter the developmental fate of flank cells to become limb cells. The present experimental system will contribute to a further elucidation on how the limb is formed.

Animals

A fibrillar collagen gene, Col11a1, is essential for skeletal morphogenesis.

Mice that are homozygous for the autosomal recessive chondrodysplasia (cho) mutation die at birth with abnormalities in cartilage of limbs, ribs, mandible, and trachea. Limb bones of newborn cho/cho mice are wider at the metaphyses than normal bones and only about half the normal length. By linkage analysis, the cho gene and the gene encoding the alpha 1 (XI) chain of cartilage collagen XI were mapped to the same region of chromosome 3. Deletion of a cytidine residue about 570 nt downstream of the translation initiation codon in cho alpha 1 (XI) mRNA causes a reading frame shift and introduces a premature stop codon. The data demonstrate that collagen XI is essential for normal formation of cartilage collagen fibrils and the cohesive properties of cartilage. The results also suggest that the normal differentiation and spatial organization of growth plate chondrocytes is critially dependent on the presence of type XI collagen in cartilage extracellular matrix.

Amino Acid Sequence

Structural and functional analysis of the promoter of the human alpha 1(XI) collagen gene.

In order to eventually elucidate the mechanisms regulating alpha 1(XI) collagen expression in cartilaginous and non-cartilaginous tissues, we performed an initial analysis of the structural-functional features of the promoter of the human gene (COL11A1). After cloning and sequencing the 5' portion of COL11A1, primer extension and nuclease protection assays identified several minor transcriptional start sites clustered around a major one located 318 base pairs from the ATG codon. Consistent with this finding, analysis of the upstream sequence revealed the absence of a TATA motif and the presence of several GC boxes. Transient transfection experiments delineated the smallest promoter sequence directing relatively high expression of a reporter gene in a cell type-specific manner. Nine nuclear protein-bound areas were located within this promoter sequence of the COL11A1 gene. Sequence homologies suggested that the majority of the footprints correspond to potential binding sites for ubiquitous nuclear proteins, such as AP2 and Sp1. Additional experimental evidence indicated that one of the protected areas may bind a transcriptional complex that is identical or closely related to the one that regulates tissue specificity in the coordinately expressed alpha 2(V) collagen gene.

Animals

Involvement of the Sonic hedgehog gene in chick feather formation.

To elucidate the molecular mechanisms of chick feather formation, we observed expression patterns of the Sonic hedgehog (Shh) gene, which is one of the vertebrate homologs of the Drosophila segment polarity gene, hedgehog, and encodes a signaling molecule functioning in limb pattern formation and motor neuron induction. We found that the Shh gene is also expressed in the apical region of the feather placodes and then in nine to eleven longitudinal stripes along feather filaments. The stripe was found to correspond to one of the outer marginal zones of each barb ridge, termed the zone of Shh expression. No significant expression signal was detected in the scale bud of developing legs. Thus, Shh is likely to function as an epithelial signaling molecule in epithelio-mesenchymal interaction during feather formation. Furthermore, since genes of bone morphogenetic protein-2 (BMP-2) and fibroblast growth factor-4 (FGF-4) are coexpressed with Shh during feather formation as observed in limb morphogenesis, interactions among FGF-4, Shh and BMP-2 may be involved in formation of feather filaments and barbs in a similar fashion as elucidated in limb pattern formation.

Animals

Analysis of proliferative activity using antiproliferating cell nuclear antigen antibody in colorectal cancer.

Using anti-PCNA antibody (PC10), an immunohistochemical study of the expression of PCNA in formalin-fixed and paraffin-embedded materials of colorectal cancer patients was performed and correlation of PCNA expression with clinicopathological findings and DNA ploidy pattern was studied. PCNA labeling rate (PCNA LR) was estimated in the advancing margin of the tumor and ranged from 23.8% to 77.9%. There was a significant difference in lymphatic vessel invasion, liver metastasis and Dukes' stage between the groups with high (> 48.7) and low (< 48.7) PCNA LR (P < 0.05). No differences were seen in tumor size, histological type, lymph node metastasis or DNA ploidy pattern. In patients with younger age, infiltration to neighboring organs, a high degree of venous invasion, and peritoneal dissemination, the frequency of high PCNA LR tended to be higher (P < 0.1). The results above suggest that a high proliferative activity as defined by evaluation of the PCNA LR at the advancing margin of the tumor may be one of the parameters of malignant potential and helpful as a predictor of liver metastasis in colorectal cancer.

Antibodies, Monoclonal

31P NMR study of acute toxic effects of cadmium chloride on rat liver.

In this study, acute effects of cadmium ions (Cd2+) on energy metabolism in rat livers were analyzed in vivo after intravenous administration using 31P NMR. Both inorganic phosphate (Pi) and nucleotide triphosphate (NTP) peaks of in vivo Cd-treated livers gradually decreased over a 6-h period. In the extract, NTP peaks in Cd-treated livers were lower, as in the in vivo experiments, but the Pi peak was significantly higher than the control. The apparent decrease in Pi in in vivo liver treated with Cd could be caused by the reduced visibility of Pi because of its uptake into mitochondria from cytoplasm, accompanied by the uncoupling of oxidative phosphorylation by Cd2+. These results indicated that total Pi in the hepatocytes increases after Cd administration. However, only 10% of Pi was visible in Cd-treated livers in vivo, whereas 34% of Pi was visible in controls. Significant increases in phosphocholine and glycerophosphocholine were also observed in extracts of Cd-treated livers.

Acute Disease

Cerebral MRI and spectroscopy in Sjögren-Larsson syndrome: case report.

We report MRI and MRS of the brain in a patient with Sjögren-Larsson syndrome (SLS) in whom fatty alcohol oxidoreductase (FAO) deficiency has been verified. MRI showed periventricular lesions, high intensity on T2-weighted and low intensity on T1-weighted images at trigones of the lateral ventricles. 1H-MRS of these lesions revealed high lipid and low N-acetyl aspartate peaks. We presume such lipids in periventricular lesions with high T2 signal may be pathognomonic of SLS.

Adult

Intracerebral hematoma in patients with ruptured cerebral aneurysms.

BACKGROUND: Intracerebral hematoma from ruptured aneurysms is one of the unfavorable factors for outcome in patients with subarachnoid hemorrhage. In this study, the clinical characteristics of intracerebral hematoma in patients with ruptured aneurysms were examined. METHODS: The subjects were 512 patients who had been admitted by day 3 after aneurysmal rupture without episodes of rebleeding before the initial computed tomography (CT) scan. They were divided into two groups according to the findings of initial CT; groups 1 and 2 comprised patients with and without intracerebral hematoma, respectively. RESULTS: Of the 512 patients, intracerebral hematoma was observed in 98 (19%). The incidence of intracerebral hematoma was higher in patients with distal anterior cerebral and middle cerebral artery aneurysms, compared with those at other sites (both, p < 0.01). Interhemispheric, callosal, and temporal lobe/sylvian hematomas were observed more frequently in patients with anterior communicating, distal anterior cerebral, and middle cerebral artery aneurysms, respectively, than in those with aneurysms at other sites. The incidence of rebleeding was 22% in group 1 and 14% in group 2 (p < 0.05). Clinical grades on admission were higher and outcome at 6 months after onset was less favorable in group 1 than in group 2 (both, p < 0.01). The larger the intracerebral hematoma, the higher was the clinical grade and the less favorable the outcome. However, when comparing management and surgical outcome under the same clinical grades, there was no significant difference between the two groups. CONCLUSIONS: There was a close correlation between the site of hematoma and that of the ruptured aneurysm. Poor outcome in patients with intracerebral hematoma seems to be related to severity of clinical grade on admission.

Adult

Prevention of autoimmune symptoms in autoimmune-prone mice by elimination of B-1 cells.

Our recent studies on an autoantibody-transgenic mouse line demonstrated that peritoneal B-1 cells are responsible for autoimmune symptoms. However, whether B-1 cells in the peritoneum are generally involved in the pathogenesis of autoimmune disease remains controversial. To test the possible involvement of peritoneal B-1 cells in autoimmune symptoms of autoimmune-prone NZB mice, we eliminated the peritoneal cells by hypotonic shock with repeated i.p. injection of distilled water every 7 days into neonatal or 8-week-old NZB mice. By this treatment, B-1 cells, which self-renew within the peritoneal cavity, are expected to be preferentially eliminated, while other peritoneal cells can be easily supplied from bone marrows after this treatment. Indeed, in distilled water-treated old NZB mice, the number of B-1 cells decreased in spleen as well as in lamina propria of the gut but the numbers of conventional B cells and T cells did not change. Moreover, the production of autoantibodies against erythrocytes significantly decreased and the occurrence of autoimmune hemolytic anemia was reduced in 12-month-old treated NZB mice. Similarly, the elimination of peritoneal cells of NZB/NZW (NZB/W) F1 mice by water injection decreased anti-DNA IgG antibodies in the sera and reduced the pathological changes of the kidney. These results suggest that peritoneal B-1 cells may be a source of autoantibody-producing cells in autoimmune diseases of NZB and NZB/W F1 mice.

Anemia, Hemolytic, Autoimmune

The mRNA for alpha 1(XIX) collagen chain, a new member of FACITs, contains a long unusual 3' untranslated region and displays many unique splicing variants.

We have isolated cDNAs and completed for the first time the primary structure for a novel collagenous chain that was partially characterized earlier and named alpha 1(Y) chain [Yoshioka, H. et al. (1992) Genomics 13, 884-886]. The size of the coding region was unexpectedly small compared with the length of the mRNA (> 10 kb), owing to the presence of a long 3' untranslated region (> 5 kb). The predicted polypeptide contained 1,142 amino acid residues with a 23-residue signal peptide consisting of 5 collagenous domains of 70-224 residues in length, interspersed and flanked with 6 noncollagenous (NC) domains. The primary structure is distinct from those of the 32 known collagen alpha-chains of types I through XVIII. Therefore, we designate this newly discovered collagen chain the alpha 1 chain of type XIX collagen. Sequence analysis suggested that this chain belongs to the recently discovered group of collagens known as FACITs (fibril associated collagens with interrupted triple-helices). Northern blotting analysis demonstrated hybridization of the cDNA to a large mRNA species (> 10 kb) extracted from a rhabdomyosarcoma cell line (CCL 136). We also isolated numerous truncated cDNA clones of which the 3' parts were different from the "proto" type of the mRNA of > 10-kb size. Sequence comparison between cDNAs and corresponding genomic DNA fragments indicated that unusual splicing events occurred through insufficient recognition at acceptor sites. Expression of the gene was extremely infrequent in the rhabdomyosarcoma cell line; it could be restricted to certain animal tissues both temporally and spatially during early development.

Alleles

Shotgun wounds and pellet emboli: case reports and review of the literature.

Shotguns produce extensive tissue damage when victims are injured at close range. Arterio-arterial and veno-venous pellet emboli may result when major vessels or their tributaries are injured. Management of the pellet embolus is dictated by whether it is symptomatic or asymptomatic. Regardless of symptoms, the embolus is a sign of possible major vascular or venous tributary injury. We recently managed two cases of pellet emboli produced by close range shotgun wounds. This report describes our management of these injuries and collectively reviews the past 30 years of literature on shotgun wounds with arterio-arterial and veno-venous pellet emboli. Recommendations for management are suggested from collating the cases reported in the literature.

Adult

Fat tissue: relationship between chemical shift and magnetization transfer.

The middle calves of the legs of 25 volunteers (12 men and 13 women, aged 18-25 years), as well as phantoms, were examined with 1.5-T magnetic resonance imaging. Gradient-recalled technique was combined with magnetization transfer contrast (MTC) and chemical shift imaging. Periodic oscillation of the MTC effect in fat tissue with variation of echo time was identified at the frequency corresponding to the water-fat chemical shift difference.

Adipose Tissue

Effects of hypercapnia on ECoG and oxidative metabolism in neonatal dog brain.

The effects of hypercapnia on cerebral electrical activity and mitochondrial oxidative phosphorylation were studied in the anesthetized neonatal dog by using the electrocorticogram (ECoG) and 31P-magnetic resonance spectroscopy. Three levels of hypercapnia with arterial PCO2 values of approximately 70, 100, and 140 Torr reduced the intracellular pH of the brain from 7.11 to 6.99, 6.87, and 6.76, respectively. These levels of hypercapnia also reduced ADP concentration ([ADP]) from 21.5 to 18.1, 14.8, and 12.9 microM as well as the average ECoG power output by 20, 30, and 40%. A Michaelis-Menten relationship for the mitochondrial respiratory enzymes was fitted with [ADP] and the change in the average ECoG. The result suggests that mitochondrial respiration is regulated by [ADP] and that the in vivo Michaelis-Menten constant for ADP was 21 microM, a value close to the in vitro value. The mitochondrial maximal reaction velocity was reduced by only 10% during hypercapnia and showed no relationship with the degree of acidosis, suggesting that mitochondrial respiratory enzymes are not responsible for the inhibition of the brain electrical activity.

Adenosine Diphosphate

[Metabolic and functional magnetic resonance imaging of the brain: clinical application to pediatric brain diseases].

We have developed 1H-chemical shift imaging (CSI) and functional magnetic resonance imaging (FMRI) methods on a clinical MRI system, in which metabolic and functional information can be obtained from the brain. 31P- and 1H-CSI are in clinical use. Using 1H-CSI, the peaks of N-acetylaspartate (NAA), choline (Cho) and creatine (Cr) are clearly detected in multiple small voxels. In normal children, the ratio of NAA/Cho increased after birth in different manners in different parts of the brain. The peak of NAA decreased in some disorders with brain damage or neuronal immaturity. It has been shown recently that functional activation of the cortex can be visualized with MR imaging with a blood oxygen level dependent (BOLD) effect. At an activated area, the ratio of deoxyhemoglobin to oxyhemoglobin decreased in the capillary and venous beds. Therefore, with a decrease of the effect of the T2 susceptibility from deoxyhemoglobin, the signal intensity of the activated area increased. CSI and FMRI have a unique possibility in the field of non-invasive brain analysis.

Adolescent

Magnetization transfer contrast on gradient echo MR imaging of the temporomandibular joint.

Thirty-nine temporomandibular joints (TMJ) from 20 patients with suspected internal derangements were imaged by a 1.5 T MR imager. The on-resonance binomial magnetization transfer contrast (MTC) pulse was applied to gradient echo images with a dual receiver coil (9 s/section). With the use of an opening device, a series of sequential images were obtained at increments of mouth opening and closing. The tissue signal intensities with (Ms) and without (Mo) MTC were measured and subjective image analysis was performed. Compared with the standard images, MTC technique provided selective signal suppression of disks. The average of Ms/Mo ratio of the disks (0.56) was lower than that of the retrodiskal pad (0.79) and of the effusion (0.89). With MTC technique, fluid conspicuity was superior to standard image. However, no significant superiority was found in disk definition subjectivity.

Adolescent