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Biomedical subjects

Y Ueda

Publications and source records attributed to Y Ueda.

At least 19 recordsLinked to original sources

The effects of a unique D-loop structure of a minor tRNA(UUALeu) from Streptomyces on its structural stability and amino acid accepting activity.

Streptomyces bldA gene, which encodes a tRNA corresponding to a very minor leucine codon, UUA, regulates pleiotropic gene expression which is involved in sporulation and secondary metabolism. The unique structural feature of this tRNA is the lack of GG sequence in dihydrouridine loop (D-loop) that generally is conserved in tRNAs involved in cytoplasmic protein biosynthesis. In order to investigate the relationship between the D-loop structure and the stability and leucine accepting activity of this tRNA, the wild and D-loop mutant tRNA transcripts were constructed with T7 RNA polymerase in vitro. The wild type tRNA(UUALeu) showed the structural stability and leucine accepting activity at physiological temperature for Streptomyces. The E.coli type D-loop mutant, which has a larger loop size and contains a GG doublet, exhibited increased thermostability. The kinetical analyses of the aminoacylation reaction of tRNA(UUALeu) with S.lividans and E.coli leucyl-tRNA synthetase (LeuRS) suggest there is a unique recognition mechanism of Streptomyces LeuRS toward tRNA(UUALeu).

Base Sequence

Synthesis and properties of mirror-image DNA.

We have investigated the conformations of the hexadeoxyribonucleotide, L-d(CGCGCG) composed of L-deoxyribose, the mirror image molecule of natural D-deoxyribose. In this paper, we report the synthesis of four L-deoxynucleosides and the L-oligonucleotide-ethidium bromide interactions. The L-deoxyribose synthon 9 was synthesized from L-arabinose with an over all yield of 28.5% via the Barton-McCombie reaction. The L-deoxynucleosides were obtained by a glycosylation of appropriate nucleobase derivatives with the 1-chloro sugar 9. After derivatization to nucleoside phosphoramidites, L-deoxycytidine and L-deoxyguanosine were incorporated into a hexadeoxynucleotide, L-d(CGCGCG) by a solid-phase beta-cyanoethylphosphoramidite method. This L-hexanucleotide was resistant to digestion with nuclease P1. The conformations of L-d(CGCGCG) were an exact mirror image of that of the corresponding natural one as described previously, and the conformations of the L-d(CGCGCG)-ethidium bromide complex were also the mirror images of those of the D-d(CGCGCG)-ethidium bromide complex under both low and high salt conditions. These results suggest that ethidium bromide prefers not a right-handed helical sense, but the base-base stacking geometry of the B-form rather than that of the Z-form. Thus, L-DNA would be a useful tool for studying DNA-drug interactions.

Chromatography, High Pressure Liquid

In vivo calcium elevations in thymocytes with T cell receptors that are specific for self ligands.

Selection of the T cell receptor (TCR) repertoire in the thymus probably involves TCR-mediated signals transduced in developing thymocytes after interaction with thymic stromal cells bearing self ligands. TCR-transduced signals should have identifiable consequences that would distinguish thymocytes whose TCRs have been engaged by self ligands from those whose TCRs have not. Among thymocytes expressing a transgenic TCR of defined specificity, a large number had elevated intracellular calcium concentrations but only when resident in a negatively selecting thymus in which their self ligand was expressed. Thus, developing thymocytes are stimulated by endogenous ligands in vivo to mobilize intracellular calcium, and increased intracellular calcium concentrations may reflect the consequences of intrathymic signaling associated with thymic negative selection.

Animals

Co-localized but target-unrelated expression of vasoactive intestinal polypeptide and galanin in rat dorsal root ganglion neurons after peripheral nerve crush injury.

Expression of vasoactive intestinal polypeptide (VIP) and galanin in dorsal root ganglion (DRG) neurons is known to be induced by peripheral nerve injury. We investigated (1) whether VIP and galanin were co-expressed by DRG neurons and (2) whether such neurons innervated specified peripheral targets (visceral, cutaneous or muscular). An antibody to the 200 kDa neurofilament subunit (NF200) was used as a marker for large type-A cells in the DRG. VIP and galanin were respectively observed in 22% and 67% of DRG neurons at the L5 spinal level after crushing of the sciatic nerve. Most VIP-containing neurons were small type-B cells (about 90%) and approximately 95% of VIP-containing neurons also showed galanin-like immunoreactivity. Galanin was expressed by both large type-A and small type-B cells. Immunocytochemistry combined with a retrograde tracer revealed that about 70-80% of the small type-B cells in each sensory division displayed VIP-like immunoreactivity, and that most of the tracer-labeled neurons also expressed galanin. These findings suggest that the expression VIP and/or galanin in response to peripheral nerve crush injury is a property common to visceral, cutaneous and muscular sensory neurons.

Animals

Neuronal survival and neurite extension supported by astrocytes co-cultured in transwells.

The influence of astrocytes on the development of cerebral cortical neurons was studied in a co-culture system using transwells with chemically defined medium. Cerebral cortical neurons from 15- or 16-day-old mouse embryo were cultured in the lower wells which were separated by a porous membrane from the upper transwells where cerebral cortical astrocytes from newborn mouse were cultured. Neurons co-cultured with astrocytes for 7 days formed a network-like web and maintained a slightly better survival from 4 to 7 days. However, neurons cultured in conditioned medium obtained from astrocytes did not form any network after 7 days even though they maintained a better cell survival at 4 days.

Animals

Axonal blockade induces the expression of vasoactive intestinal polypeptide and galanin in rat dorsal root ganglion neurons.

Nerve growth factor (NGF) undergoes retrograde transport from peripheral target organs, and has been recently reported to regulate the production of some neuropeptides in dorsal root ganglion (DRG) neurons. Therefore, to ascertain whether or not the expression of calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP), and galanin was regulated by the retrograde transport of factors such as NGF, we carried out an immunocytochemical analysis using vinblastine as an axonal transport blocker and a monoclonal antibody to the NGF receptor (NGFR) as a marker of NGF-responsive neurons. The percentage of CGRP-containing DRG neurons (L5) was decreased by sciatic nerve transection or by the application of higher doses of vinblastine (0.3-0.6 mM) to the sciatic nerve. VIP and galanin were expressed in some DRG neurons after the application of a low dose of vinblastine (0.15 mM), which can block axonal flow without causing neuronal damage. The expression of these peptides was not affected by dorsal rhizotomy. About 70% of the CGRP-containing neurons also expressed NGFR, while most of the VIP-containing or galanin-containing neurons lacked NGFR. These findings indicate that the depletion of peripheral target-derived neurotrophic factor(s) other than NGF by axonal blockade may induce the gene expression of VIP and galanin.

Animals

ADF (adult T cell leukemia-derived factor)/human thioredoxin and viral infection: possible new therapeutic approach.

ADF (adult T-cell leukemia-derived factor), originally defined as an inducer of interleukin 2 receptor/alpha (IL-2R/alpha), is a homologue of thioredoxin. ADF is constitutively produced by human lymphoid cell lines transformed by human T-lymphotropic virus type I (HTLV-I) or Epstein-Barr virus (EBV). ADF augments the proliferation of HTLV-I and EBV transformed cells as an autocrine growth factor. These data are indicative of the possible involvement of ADF in virus-related transformation of cells and their autocrine growth. On the other hand, thioredoxin contains a redox active disulfide and has a reducing activity in the presence of thioredoxin reductase and NADPH. To clarify the role of ADF/thioredoxin system in the viral transformation, we tested the effect of 13-cis-retinoic acid (RA), which is a competitive inhibitor of thioredoxin reductase, on the growth of ADF high producing cells. The expression of IL-2R/alpha on HTLV-I (+) cells was suppressed by RA. RA dose-dependently reduced the cell number and viability of ADF high producing lymphoid cells. Moreover, it had a suppressive effect on the proliferation of ADF high producing cells. It is suggested that RA has an inhibitory effect on the activation and the growth of cells producing ADF and that inhibition of the ADF/thioredoxin system may be a new therapeutic approach for retrovirus-related disorders.

Amino Acid Sequence

Case report 699. Primary localized skeletal cryptococcosis (torulosis) of the right tibia at its proximal end.

The case of a 58-year-old man with a normal immune status with primary localized skeletal cryptococcosis of the right tibia was presented. Radiologically it appeared as an osteolytic lesion with marginally indistinct borders in the epimetaphyseal region of the proximal tibia. Histologically, necrotizing granulomatous inflammation was associated with a prominent fibrohistiocytic reaction. The presence of cryptococci was confirmed by special stains. Clinicopathological features and the therapeutic aspects of this rare infective bone lesion were discussed. It is uncommon today, particularly in developed countries, to encounter a localized skeletal fungal infection. Therefore, radiologists and pathologists generally may be unfamiliar with the appearance of the lesion and tend not to consider the possibility of fungal osseous disease in the differential diagnosis. Unless it is remembered that fungal infection may cause a localized bone lesion with varied histological responses, the diagnosis may be overlooked, and bacteriologic proof may not be sought.

Bone Diseases

Effect of increase in heart rate on interatrial shunt in atrial septal defect.

The effect of increases in heart rate by atrial pacing was investigated in 20 children [mean 9 +/- 4 (SD) years] with atrial septal defect. Systemic blood flow increased by 13 +/- 10 and 27 +/- 22% with a 25 and 50% increase in heart rate, respectively. Pulmonary blood flow, however, remained unaltered. Thus, rapid pacing decreased the pulmonary to systemic blood flow ratio significantly (-15 +/- 10 and -22 +/- 13%, respectively). This hemodynamic alteration was attributed to the difference in diastolic distensibility (compliance) between the left and right ventricles, and the ratio of right ventricular to left ventricular compliance (defined by dV/dP) near the end-diastolic pressure was estimated to be 6.5 +/- 4.2. It is suspected that the increase in heart rate may contribute to the lowering of pulmonary to systemic flow ratio during exercise in children with atrial septal defect.

Adolescent

Cellular variant of extraskeletal myxoid chondrosarcoma of abdominal wall--a case report with comparative immunohistochemical study on cartilaginous collagenous proteins in various myxoid mesenchymal tumors.

A rare cellular variant of recurrent extraskeletal myxoid chondrosarcoma occurring in the right lower abdominal wall of a 70-year-old man, is presented with emphasis on a characteristic distribution pattern of cartilaginous collagen proteins in the stroma. While the primary and the first recurrent tumors showed the typical histology of extraskeletal myxoid chondrosarcoma, the later tumor, which recurred 14 years after the first resection, comprised mostly compact nodules of proliferating anaplastic cells with little mucoid stroma. Some areas presented a hemangiopericytomatous pattern. A few nodules possessed abundant myxoid stroma. Immunohistochemically, type II collagen was demonstrated in the stroma of some cellular areas, and type VI collagen was intensely stained around individual tumor cells both in cellular and myxoid areas. In a comparative immunohistochemical study, the same distribution pattern of cartilaginous collagen proteins was observed only in skeletal myxoid chondrosarcomas, but not in other mesenchymal tumors with abundant myxoid stroma. These findings seem to support the cartilaginous nature of extraskeletal myxoid chondrosarcoma, and will facilitate the differential diagnosis of soft-tissue tumors with myxoid stroma.

Abdominal Muscles

Osteofibrous dysplasia of long bones--a reactive process to adamantinomatous tissue.

The most controversial aspect of osteofibrous dysplasia (OFD) is its possible histogenetic relationship to adamantinoma of long bone. Evidence is recently beginning to accumulate that OFD may be a reactive process to regressive adamantinoma. To verify the concept, 13 lesions of OFD were studied again by immunohistochemistry for cytokeratins of different molecular masses, as well as by conventional stainings. In addition, 2 adamantinomas and 6 fibrous dysplasias of the tibia were studied for reference. A small number of spindle- or ovoid-shaped cells scattered individually in the fibro-osseous stroma showed positive reactions for cytokeratins of 55-57 kDa in 2 lesions, and for those of 45-56.5 kDa in 8 lesions of 13 OFDs, although no definite epithelial island could be detected even by immunohistochemistry. Adamantinomas also showed single cytokeratin-positive cells dispersed in fibroblastic stroma, in addition to epithelial islands positive for cytokeratins of both 55-57 kDa and 45-56.5 kDa. All cases of fibrous dysplasia were negative for cytokeratins. During the observation, no case of OFDs progressed to classic adamantinoma. The present study, demonstrating the existence of an intermediate stage between "differentiated adamantinoma" and total elimination of adamantinomatous components, gives further support for the concept that OFD is a secondary reactive process to adamantinomatous tissue. In practice, the existence of single scattered cytokeratin-immunoreactive cells in otherwise typical OFDs may not indicate the truly malignant behaviour of classic adamantinoma, unless discrete epithelioid cell nests are also found.

Adolescent

Effect of chemotherapy combined with caffeine for osteosarcoma.

Nine patients with osteosarcoma were treated by chemotherapy combined with caffeine and surgery. All primary tumors showed complete histological response to preoperative chemotherapy consisting of three intraarterial infusions of cisplatin and caffeine without/with doxorubicin and two systemic high-dose methotrexate combined with vincristine. Limb-salvage surgery was performed in eight patients with marginal procedure, which led to the preservation of good limb function. Below-knee amputation was done in one patient with calcaneal osteosarcoma. There has been neither local recurrence nor lung metastasis in seven patients with conventional osteosarcoma during a median follow-up period of 28 months. Lung metastases leading to death were observed in one patient with small-cell osteosarcoma despite complete destruction of the primary tumor by preoperative chemotherapy. Chemotherapy combined with caffeine administration deserves further extensive and large-scale study in osteosarcoma.

Adolescent

Chondroblastoma of bone. A clinical, radiological, light and immunohistochemical study.

The clinical and morphological findings of 53 chondroblastomas in the files of the Bone Tumour Registry of Westphalia are presented. The mean age of all patients was 19.2 years. The male-to-female ratio was 1.5:1. Forty-two of the tumours (79.8%) were located in the long tubular bones and short tubular bones of the hands and were closely related to the growth plate. Six cases (11.3%) were found in the flat bones, 4 cases (7.5%) in the tarsal bones and 1 case (1.9%) in the craniofacial bones. The characteristic radiological feature of 44 investigated lesions was a mostly eccentric radiolucency with a geographic pattern of bone destruction and matrix calcifications. Periosteal reaction was evident in 9% of the cases. Most tumours demonstrate the typical morphological features of chondroblastoma, but 3 cases resembled a giant cell tumour. In 2 cases a haemangiopericytoma-like growth pattern was observed. Nine of the tumours had an aneurysmal bone cyst-like component. Vascular invasion was seen in 1 case. Immunohistochemically most cells in 30 of the cases and fetal chondroblasts in 3 cases were strongly positive with vimentin and S-100 protein. Collagen type II was positive in the chondroid matrix of the tumours and in fetal cartilage tissue; collagen type VI was present focally around individual tumour cells and was always seen in the chondroid matrix of the lesions and in fetal cartilage. These findings support the cartilaginous nature of these tumours. In paraffin sections, 46.6% of the cases revealed a distinct positive reaction of some tumour cells with the monoclonal cytokeratin antibody KL1 (molecular weight 55-57 kDa). Only 4 of them demonstrated a coexpression with the other monoclonal cytokeratin antibody CK (clone MNF 116, molecular weight 45-56.5 kDa). In paraffin sections all fetal chondroblasts were negative with both cytokeratin antibodies. Frozen sections of 3 tumours showed a strong positive reaction with both cytokeratin antibodies in many chondroblasts, indicating an "aberrant" cytokeratin expression. Osteoclast-like giant cells stained positive with leucocyte-common antigen (LCA) and with the macrophage-associated antibody KP1, but were negative with the other macrophage-associated antibody MAC 387. Recurrence rate was 10.7%. The clinical course of all tumours was benign.

Adolescent

Improved procedure of complex demodulation and an application to frequency analysis of sleep spindles in EEG.

Complex demodulation is a local version of harmonic analysis that enables the amplitude and phase of particular frequency components of a time series to be described as functions of time. The paper presents a computational procedure involving complex demodulation with interpolation of data in the frequency domain. A computational procedure comprising repeated use of complex demodulation is also presented. This is used to estimate the optimum choice of the demodulating frequency which considerably influences the measurement of the instantaneous amplitude and phase of the underlying process. The usefulness of this procedure is verified by computer simulation. An example of applying this procedure to the estimation of the centre and the instantaneous frequencies of sleep spindles in the EEG (electroencephalogram) is presented. By using the procedure developed here, several partially overlapping sleep spindles are detected and correctly separated. The paper also presents an approach to separating and analysing transient time series (such as overlapping sleep spindles) by using an accurate frequency processing technique.

Computer Simulation

Acute pulmonary edema after Blalock-Taussig anastomosis.

Acute pulmonary edema developed in 2 patients after a Blalock-Taussig shunt procedure. Both patients had a univentricular heart complex with pulmonary stenosis. One patient underwent a modified Blalock-Taussig shunt using a polytetrafluoroethylene tube and the other underwent a classic Blalock-Taussig anastomosis. Acute pulmonary edema after the Blalock-Taussig shunt is rare, but once it has occurred, immediate treatment is obligatory.

Acute Disease