Biomedical subjects

K Toyoshima

Publications and source records attributed to K Toyoshima.

At least 127 records · Page 7Linked to original sources

PubMed1993

The presence of vesicles containing lactate dehydrogenase in the ameloblast layer of bovine enamel organ.

Ameloblast layers were removed from bovine enamel organs, followed by the separation of the extracellular matrix-vesicle fraction after collagenase digestion. Lactate dehydrogenase (LDH)-containing vesicles were found in that fraction. LDH in these vesicles did not result from cell lysis and vesicle capture during the preparation of the fraction. The isoenzyme pattern of LDH in LDH-containing vesicles was similar to that of cytosolic LDH of ameloblasts, suggesting the presence of a mechanism for specific uptake of cytosolic LDH during the in vivo formation of the vesicles. The existence of LDH-containing vesicles in enamel organ, where mineralization had been believed not to be initiated by matrix vesicles, suggests the possibility that LDH-containing vesicles have a specific function different from that of matrix vesicles.

Ameloblasts↗

PubMed1993

Glossopharyngeal-hypoglossal nerve reflex of the frog in metamorphosis.

In adult frogs, a reflex discharge is elicited from the hypoglossal nerve by mechanical and chemical stimuli to the centre of the tongue surface and electrical stimuli to the glossopharyngeal nerve innervating tongue. However, there has been no report concerning the embryological stage at which this reflex develops. The metamorphosis of a frog is divided into three parts, premetamorphic, metamorphic and climax stage. In the climax stage, the forelimbs are lengthened and the tail gradually shortened. The glossopharyngeal-hypoglossal nerve reflex arose in the early climax stage, during which the tongue appeared on the floor of the mouth. At this stage, the conduction velocities of the first compound action potential of the glossopharyngeal and hypoglossal nerves were about 8.7 and 9.3 m/s, respectively. The latency of the reflex was long, especially the central latency. The response of the glossopharyngeal nerve and the reflex discharge from the hypoglossal nerve upon mechanical stimulation of the tongue were smaller in the tadpole than in the adult frog. As metamorphosis proceeded, the conduction velocity of these nerves increased; the latency shortened, especially the central latency; the reflex response to mechanical stimulus increased. These results suggest that, although the glossopharyngeal-hypoglossal nerve reflex is developed in the early climax stage, it is completed only in the adult frog. Therefore, adult frogs living mainly on land may reject and swallow prey by this reflective tongue movement.

Action Potentials↗

PubMed1993

The murine cot proto-oncogene: genome structure and tissue-specific expression.

We cloned and analyzed the murine cot proto-oncogene and examined its tissue-specific expression in fetal, newborn and adult mice. Genomic cot DNA consists of eight exons, spanning more than 25 kb, and all intron-exon borders are well conserved as compared to the human homolog. Analysis of the full-length cot cDNA revealed that it contained an open reading frame of 1,401 nucleotides, like human cot proto-oncogene. The sequence identity between murine and human cot gene is 84.4% at the nucleotide level and 93.9% at the deduced amino acid level. On northern blot analysis of poly (A)+ RNA, the cot message was detected at 2.9 kb in size. Expression of the cot gene was observed in many tissues from fetal to adult mice, though the level of expression was low in all tissues examined.

Amino Acid Sequence↗

PubMed1993

Construction of immunoradiometric assay for circulating c-erbB-2 protooncogene product in advanced breast cancer patients.

The human c-erbB-2 protooncogene product (erbB-2 protein) is a 185 kilodalton glycoprotein closely related to epidermal growth factor receptor protein. In this study, we measured the concentration of circulating erbB-2 protein in cancer patients by means of a new immunoradiometric assay (IRMA). Two monoclonal antibodies (MoAbs), SV2-61 gamma and 6G10, recognize erbB-2 protein but bind to separate epitopes. SV2-61 gamma was used as an immunoadsorbent and 6G10 as an 125I-labeled probe. A serum was considered positive for erbB-2 protein if the percent binding exceeded the mean of the normal group by more than 3 standard deviations. Eleven of 21 patients with advanced breast cancer and 1 of 15 with advanced gastric cancer were positive. Serum erbB-2 protein levels correlated well with the therapy and the status of the patients with breast cancer. On the contrary, all patients with advanced colon, ovarian, or pancreatic cancers, showed levels below the cut-off value. These results suggest that circulating erbB-2 protein can be measured using the newly constructed IRMA. Since c-erbB-2 protooncogene amplification and overexpression are accepted as a good marker of aggressiveness, relapsing potency, and poor prognosis, this IRMA should be a promising tool with which to help manage breast cancer patients.

Adenocarcinoma↗

PubMed1993

Association of Src-family kinase Lyn with B-cell antigen receptor.

Antigen is thought to cross-link membrane-bound immunoglobulins (Igs) of B cells, causing proliferation and differentiation or the inhibition of growth. Compelling evidence suggests that protein-tyrosine phosphorylation is involved in signal transduction for cell proliferation and differentiation. Indeed cross-linking of membrane-bound IgM (mIgM) induced a rapid increase in tyrosine phosphorylation of at least 10 distinct proteins in B cells. The Src-family protein tyrosine kinase Lyn (p56lyn and p53lyn) is expressed preferentially in B cells. The Lyn protein and its kinase activity could be coimmunoprecipitated with both IgM and IgD from detergent lysates. Cross-linking of membrane-bound IgM with antibody induced down-regulation of the Lyn protein. From these data we concluded that Lyn is physically associated with mIgs. Further evidence showed that cross-linking of mIgM induced rapid increase in the kinase activity of Lyn and association of Lyn with 85-kDa noncatalytic subunit of phosphatidylinositol 3-kinase. Thus, Lyn is likely to participate in B-cell antigen receptor-mediated signaling. As a novel signaling molecule downstream of Lyn, we identified src homology 3-containing, transcription factor-like molecule p75HS1.

Adaptor Proteins, Signal Transducing↗

PubMed1993

Occurrence of melanosome-containing Merkel cells in mammalian oral mucosa.

The occurrence and fine structure of Merkel cells with melanosomes in the labial epithelium of the dog and cat were studied by electron microscopy. These melanosomes were polarized in the Merkel cell cytoplasm opposite to the major aggregation of dense-cored granules and the closely abutting nerve terminal. None of melanosomes was found in the Golgi complex region where a few immature dense-cored granules were observed. This may suggest that the melanosomes are not produced in the Merkel cell, but are taken from the melanocytes like in keratinocytes. The possible phagocytotic ability of Merkel cells is discussed.

Animals↗

PubMed1993

Are Merkel and Grandry cells two varieties of the same cell in birds?

For over a century it has been held that the Grandry corpuscle and the Merkel corpuscle are unique sensory organs in aquatic and nonaquatic birds, respectively. In other words, the Grandry and the Merkel cells are two varieties of the same receptor cell and never coexist together. Contrary to what has been believed, we found unmistakable Merkel cells in addition to Grandry cells in the lingual mucosa of the duck, Anas platyrhynchos var. domestica, an unprecedented observation in an aquatic bird. This study concerns the fine structure of these cells in the tongue of the duck. Although some ultrastructural features are shared by the Grandry and the Merkel cells, i.e., the presence of numerous dense-cored granules and microvillous projections at the cell surface, the morphology of these two types of cells clearly differ, especially in their size. Cells suggestive of stages intermediate between these two cells have never been recognizable. The present study indicates that the Grandry and the Merkel cells are not two varieties of the same cell in birds. It is suggested that the Merkel cell may exist in every vertebrate species, while the Grandry cell is unique to aquatic birds.

Animals↗

PubMed1993

The occurrence and fine structure of Merkel cells in the lingual epithelium of the turtle, Clemmys japonica.

Electron microscopic examination of the lingual mucosa of the turtle, Clemmys japonica, revealed the occurrence of Merkel cells that shared many morphological features with Merkel cells in other vertebrates. Merkel cells were located exclusively in the basal portion of the epithelium near the taste buds. We also found occasional Merkel cells devoid of nerve contact. Nerve terminals approaching these non-innervated Merkel cells were occasionally found in the connective tissue just beneath these cells.

Animals↗

PubMed1993

Research note: effect of tu-chung leaf (Eucommia ulmoides) on egg production performance, egg quality, and fat metabolism in laying hens at a late production stage.

The effects of tu-chung leaf (Eucommia ulmoides) supplementation on egg production performance, egg quality, and fat metabolism were investigated in laying hens at a late production stage. White Leghorn laying hens at 20 mo of age were provided ad libitum access to a practical diet with or without tu-chung leaf supplement for 3 mo. During the experimental period, egg production rate, egg weight, and feed intake were recorded every day. Egg samples from the last 3 days were collected to measure Haugh units and cholesterol concentration in the egg yolk. Plasma cholesterol concentration and abdominal fat weight were also determined. The results showed that no significant improvement in egg production was observed with tu-chung leaf addition. No significant effect of tu-chung was observed on Haugh units, plasma cholesterol, or egg yolk cholesterol, whereas abdominal fat weight was greater in hens fed the tu-chung diet than in those given the unsupplemented control diet. It was concluded that the beneficial effects of tu-chung leaf supplementation on egg production performance and egg quality were ambiguous and of little practical significance.

Animals↗

PubMed1993

[Lymphocyte subpopulation in asthmatic children with chronic bronchial obstruction].

Lymphocyte subpopulations in peripheral blood were examined in asthmatic children, using fluorescent monoclonal antibodies. The patients were divided into three groups: Group I, no attack (FEV1.0% > or = 80% group), Group II, no attack (FEV1.0% < 80% group) and Group III, asthmatic attack group. The percentage of CD3HLADR/CD3 in the Group II and Group III were 5.58 +/- 3.59 and 5.44 +/- 4.34, respectively, which were significantly higher than that in the Group I subjects (2.70 +/- 2.17). This finding suggests that the patients with long lasting bronchial obstruction may have chronic inflammation in the airway even in the remission stage of asthmatic attacks.

Adolescent↗

PubMed1993

[Efficacy of long-term beclomethasone dipropionate inhalation therapy in pediatric patients with asthma, and its effect on height growth and adrenocortical functions].

Seventy-eight pediatric patients with moderate to severe chronic asthma, aged 6 to 16 years, who failed to respond well to inhaled DSCG and theophylline RTC, were treated with beclomethasone dipropionate inhaler (BDI) for 0.5 to 10 years (mean: 4.2 +/- 2.4 years). The efficacy rate gradually increased with time: 61.1% at one year after the start of the treatment and 89.5% at three years. Long-term BDI therapy over five years did not cause suppression of height growth or adrenocortical functions (early morning cortisol level and rapid ACTH test). However, the safety of long-term BDI therapy in children still needs to be studied more thoroughly and established. Further investigation will be necessary since some cases did not respond to BDI therapy while others died of exacerbated asthmatic symptoms soon after the start of BDI therapy.

Administration, Inhalation↗

PubMed1992

An immunohistologic evaluation of C-erbB-2 gene product in patients with urinary bladder carcinoma.

BACKGROUND: Amplification or overexpression of the c-erbB-2 gene have been reported to correlate with poor patient prognosis in human breast, gastric, and ovarian cancer. Recently, the c-erbB-2 gene product was found to be expressed frequently in the urinary bladder carcinoma. In the current study, the presence of the c-erbB-2 gene product in urinary bladder carcinomas was compared with patient outcome to evaluate whether c-erbB-2 gene product could identify a subset of patients who are destined to have a poor prognosis. METHODS: Immunohistologic study of the c-erbB-2 gene product was done in formaldehyde-fixed paraffin-embedded tissue specimens obtained from 88 transitional cell carcinomas of the human urinary bladder. Eighty-three patients who underwent complete tumor resection by total cystoprostatectomy (30 patients) or by bladder-preserving operations such as transurethral surgery (50 patients) or partial cystectomy (3 patients) entered a follow-up study. The other five patients did not enter the follow-up study because of lost follow-up (2 patients) or distant metastasis at the time of surgery. RESULTS: The c-erbB-2 gene product was expressed in 23 of 88 patients (26%), showing an increase in the expression rate corresponding to the advancement of tumor grade (P < 0.05) and tumor stage (P < 0.2). The 5-year disease-free survival rate was 48.5% for patients with c-erbB-2 negative tumors versus 9.7% for those with c-erbB-2 positive tumors (P < 0.01). The 5-year actuarial survival rate was 65.5% for patients with c-erbB-2 negative tumors versus 41.8% for those with c-erbB-2 positive tumors (P < 0.05). Multivariate analysis using Cox regression model showed that the c-erbB-2 gene product tissue status was a significant prognostic factor independent of grade and stage of the tumor. CONCLUSIONS: The results suggest that the c-erbB-2 gene product could be a tumor marker to identify a malignant subgroup in bladder carcinomas.

Adult↗

PubMed1992

Phosphorylation of the retinoblastoma protein by cdk2.

The retinoblastoma gene product (the RB protein) is phosphorylated in a cell cycle-dependent manner and this modification is believed to be important for cells to progress through the cell cycle. We found that purified cdk2 (cyclin-dependent kinase/cell division kinase 2) can phosphorylate the RB protein in vitro at the sites phosphorylated in the cell. The timing of activation of cdk2 in the cell cycle was similar to that of the onset of phosphorylation of the RB protein. The kinase coprecipitated with the RB protein also exhibited a similar substrate specificity to cdk2 and a similar time course of activation during the cell cycle. We further showed that cdk2 formed a complex with the RB protein in vitro and that its formation was not competitively inhibited by the simian virus 40 large T antigen. These observations suggest that cdk2 or a cdk2-related protein is involved in the cell cycle-dependent phosphorylation of the RB protein.

CDC2-CDC28 Kinases↗

PubMed1992

Survey of 1120 Japanese women with a history of recurrent spontaneous abortions.

Our recent 7-year clinical survey showed that among the 1120 women with repeated spontaneous abortions registered in this clinic, 2898 out of a total of 3216 pregnancies (90.1%) had terminated in spontaneous abortion. Among these wastages, 84.2% occurred before 12 weeks of gestation, and 11.1 percent occurred between 12 and 15 weeks. Through routine examination of reproductive wastage, 82 (9.9%) of the 825 Japanese couples examined were shown to have either a chromosomal abnormality or normal variants in the wife and/or husband, thus demonstrating no racial difference in the incidence of chromosomal abnormalities in infertile patients in comparison with studies performed in other countries. One hundred and forty-seven congenital uterine anomalies (14.7%) were found in 1000 hysterosalpingographies, and 12 of 148 examined females were positive for anti-cardiolipin antibody. 393 other females with no major abnormalities likely to induce spontaneous abortions were indicated for immunotherapy. Ample time spent on genetic counseling prevented further reproductive wastage, and ideal metroplasty resulted in a successful post-operative pregnancy course in more than 85% of cases. Immunosuppressant and anticoagulant therapy decreased the serum titer of anti-cardiolipin antibody, enabling pregnancies to be maintained to term. Immunotherapy utilizing the husband's lymphocytes also brought more than 80% of pregnancies to successful completion, with 200 deliveries achieved with this therapy. In contrast, 64.1% of pregnancies again terminated spontaneously in patients who were indicated for immunotherapy but did not receive treatment. The findings of the present study suggest that the causes of reproductive wastage, especially the etiology of early recurrent spontaneous abortion, are complex.(ABSTRACT TRUNCATED AT 250 WORDS)

Abortion, Habitual↗

PubMed1992

Ear3/COUP-TF binds most tightly to a response element with tandem repeat separated by one nucleotide.

Ear3/COUP-TF belongs to the steroid/thyroid hormone receptor superfamily of nuclear transcription factors. Previous studies have identified six target response elements to which the factors of the family bind with various affinities. They are palindromic pair of hexameric half site (Thyroid hormone Response Element; TRE, Estrogen Response Element; ERE, Glucocorticoid Response Element; GRE) or directly repeated half sites [Direct Repeat (DR) 3, DR4, DR5]. In this report, these six elements and three additional putative elements, DR0, DR1, and DR2, which are directly repeated half sites separated by 0, 1 and 2 nucleotides, respectively, were tested for binding to Ear3/COUP-TF protein employing gel retardation method. Among the palindromic series of response elements, TRE showed the highest affinity, ERE showed second highest, and no binding was observed for GRE. Among six direct repeat response elements, DR1 showed the highest affinity. Comparison of binding affinities between DR1 and TRE revealed that the DR1 has the highest affinity of all the nine response elements tested. The consensus sequence of the DR1 element resembles well the COUP sequence found in the upstream promoter of chicken ovalbumin gene, which has been shown to serve as a natural response element for Ear3/COUP-TF. Retinoid X receptor (RXR), another member of the receptor family, reportedly utilizes DR1 sequence as its response element, suggesting that Ear3/COUP-TF could regulate the expression of the same subsets of genes as RXR does.

Base Sequence↗

PubMed1992

Activation of Src-like protein-tyrosine kinase Lyn and its association with phosphatidylinositol 3-kinase upon B-cell antigen receptor-mediated signaling.

Crosslinking of membrane-bound immunoglobulins, which are B-cell antigen receptors, causes proliferation and differentiation of B cells or the inhibition of their growth. The receptor-mediated signaling involves tyrosine phosphorylation of cellular proteins. The Src-like protein-tyrosine kinase Lyn is expressed preferentially in B cells and is an intracytoplasmic constituent of the B-cell antigen receptor complex. Crosslinking of membrane-bound immunoglobulin M with antibody induced rapid increases in the kinase activities of Lyn and Lyn-associated phosphatidylinositol 3-kinase. Crosslinking of B-cell antigen receptor also induced association of Lyn with an 85-kDa noncatalytic subunit of phosphatidylinositol 3-kinase. Thus, Lyn is functionally associated with membrane-bound immunoglobulin M and seems likely to participate in B-cell antigen receptor-mediated signaling.

B-Lymphocytes↗

PubMed1992

Protein tyrosine kinases belonging to the src family.

There are nine non-receptor-type protein tyrosine kinases that show a high level of similarity in their primary structures and in the structures of their functional domains. Together, they are called the src family. They seem to have common sites specific for oncogenic activation. Recent findings suggest that the kinases are closely associated with cell surface molecules and that they mediate extracellular signals through the activation of their tyrosine kinase activity. They appear to act more on the differentiated phenotype than in haemopoietic cell proliferation. Possible functions of the products of the lck, fyn, lyn and fgr genes in lymphocytes and monocytes are discussed.

Gene Expression Regulation, Neoplastic↗

PubMed1992

Purification and characterization of a possible protooncogene fyn product, p59fyn, from a rat brain particulate fraction.

Four tyrosine-protein kinases that reacted with antibodies specific to p62c-yes, p60c-src, p60c-src+, and p59fyn, respectively, were solubilized from a rat brain particulate fraction and separated by casein-Toyopearl column chromatography. Possible p59fyn, with a pI of 6.5, was purified 490-fold as a single 59-kDa protein band on SDS-PAGE. The purified enzyme contained almost no phosphotyrosine residues but was autophosphorylated with Mg2+. ATP exclusively at tyrosine residues, with a concomitant increase in the kinase activity toward tyrosine-glutamate (1:4) copolymers. The rate of the copolymer phosphorylation was proportional to the square of the enzyme concentration, suggesting activation through intermolecular catalysis. In the presence of Mn2+, however, the reaction showed a first-order dependence on the enzyme concentration.

Animals↗