Biomedical subjects

S Shimizu

Publications and source records attributed to S Shimizu.

At least 451 records · Page 25Linked to original sources

PubMed1996

Characterization of the gene cluster of high-molecular-mass nitrile hydratase (H-NHase) induced by its reaction product in Rhodococcus rhodochrous J1.

The 4.6-kb region 5'-upstream from the gene encoding a cobalt-containing and amide-induced high molecular mass-nitrile hydratase (H-NHase) from Rhodococcus rhodochrous J1 was found to be required for the expression of the H-NHase gene with a host-vector system in a Rhodococcus strain. Sequence analysis has revealed that there are at least five open reading frames (H-ORF1 approximately 5) in addition to H-NHase alpha- and beta-subunit genes. Deletion of H-ORF1 and H-ORF2 resulted in decrease of NHase activity, suggesting a positive regulatory role of both ORFs in the expression of the H-NHase gene. H-ORF1 showed significant similarity to a regulatory protein, AmiC, which is involved in regulation of amidase expression by binding an inducer amide in Pseudomonas aeruginosa. H-ORF4, which has been found to be uninvolved in regulation of H-NHase expression by enzyme assay for its deletion transformant and Northern blot analysis for R. rhodochrous J1, showed high similarity to transposases from insertion sequences of several bacteria. Determination of H-NHase activity and H-NHase mRNA levels in R. rhodochrous J1 has indicated that the expression of the H-NHase gene is regulated by an amide at the transcriptional level. These findings suggest the participation of H-ORF4 (IS1164) in the organization of the H-NHase gene cluster and the involvement of H-ORF1 in unusual induction mechanism, in which H-NHase is formed by amides (the products in the NHase reaction), but not by nitriles (the substrates).

Amino Acid Sequence↗

PubMed1996

Involvement of CPP32/Yama(-like) proteases in Fas-mediated apoptosis.

Fas (Apo-1/CD95) belongs to the tumor necrosis factor/nerve growth factor receptor family and transmits apoptotic signals by binding to its ligand. Interleukin-1beta-converting enzyme (ICE), which shows substantial homology to the product of the cell death gene, ced-3, of Caenorhabditis elegans, is reported to be involved in Fas-mediated apoptosis. Using two human carcinoma-derived cell lines with undetectable levels of ICE, we found that an agonistic antihuman Fas antibody induces the activation of CPP32/Yama(-like) proteases that are ICE(-like) protease family members, and that a tetrapeptide inhibitor of CPP32/Yama protease, DEVD-CHO, inhibits the Fas-mediated activation of the proteases, Fas-mediated apoptosis, and CPP32/Yama(-like) proteolytic activities in vitro. Fas-mediated apoptosis is inhibited by the CPP32/Yama inhibitor DEVD-CHO, but not by the ICE inhibitor YVAD-CHO, suggesting a dominant role for the CPP32/Yama(-like) proteases and not ICE itself in Fas-mediated apoptosis of the human carcinoma cell lines.

Amino Acid Sequence↗

PubMed1996

Subcellular localization of docosahexaenoic acid and arachidonic acid omega-hydroxylation activity in the brain, liver and colonic adenocarcinoma.

A homogenate of rat brain, rat liver or human colonic well differentiated adenocarcinoma was prepared in 250 mM sucrose isoosmolaric buffer (pH 7.6) and fractionated by differential centrifugation at 10(3), 10(4) and 10(5) g. Each precipitate or supernatant was incubated with NADPH and docosahexaenoic acid or arachidonic acid as a substrate for 30 min at 37 degrees c under aerobic conditions. omega-Hydroxydocosahexaenoic acid or omega-hydroxyeicosatetraenoic acid from an incubation mixture was detected by reversed-phase high-performance liquid chromatography-thermospray mass spectrometry with selected-ion monitoring. omega-Hydroxy polyunsaturated fatty acids were characterized by high intensity of the molecular ion (MH+) although common hydroxy polyunsaturated fatty acids were characterized by high intensity of the MH+ -H2O ion. For the rat brain, omega-hydroxylation activity (the amount of omega-hydroxy product produced in 30 min) was concentrated to a 10(3) g precipitate although the specific activity (the activity per 1 mg of protein) in the 10(3) g precipitate did not indicate superiority over other fractions. However, the specific activity of the rat brain increased on addition of a 10(4) or 10(5) g precipitate. For the rat liver, although omega-hydroxylation activity was concentrated to a 10(3) g precipitate, the specific activity was concentrated to a 10(5) g precipitate and the subcellular localization differed from that of rat brain. In the human colonic well differentiated adenocarcinoma, although omega-hydroxylation activity was relatively high in the 10(3) g supernatant, the specific activity was relatively high in the 10(3) g precipitates. These results suggest that there is a difference regarding subcellular localization of the omega-hydroxylation activity depending on the species of the organs.

Adenocarcinoma↗

PubMed1996

Histiocytoid Breast Carcinoma: A Case Report with Immunohistochemical and Ultrastructural Studies.

This is the first documented report of a case of histiocytoid breast carcinoma in Japan. The patient was a 55-year-old woman with a right breast lump. An excisional biopsy revealed proliferation of histiocyte-like cells with slightly atypical nuclei within fibrous stroma. In places, tumor cells showed an invasive pattern similar to that in invasive lobular carcinoma including a targetoid pattern, as well as a focus of in situ lobular carcinoma with transition to the histiocytoid cells. Immunohistochemical stainings of the tumor cells demonstrated positive reactions for cytokeratins and epithelial membrane antigen. Gross cystic disease fluid protein 15, an apocrine marker, was almost uniformly positive in their cytoplasm. From these results, we supposed that histiocytoid breast carcinoma is a variant of invasive lobular carcinoma. Histiocytoid breast carcinoma is easily misdiagnosed as a benign lesion such as granular cell tumor or xanthoma. Points of differential diagnosis were described and we stressed that it is most important for pathologists to keep this variant in mind in order to avoid misdiagnosis.

Journal Article↗

PubMed1996

Crystallization and preliminary X-ray diffraction study of aldehyde reductase from a red yeast, Sporobolomyces salmonicolor.

Crystals of aldehyde reductase from a red yeast, Sporobolomyces salmonicolor, have been grown from an ammonium sulfate solution, pH 7.0, by means of the vapor-diffusion procedure. The crystals belong to the hexagonal system, space group P6(1)22 or its enantiomorph, P6(5)22, with unit-cell dimensions of a = 72.2 and c = 320 A. The X-ray diffraction patterns extend to at least 2 A resolution with the use of synchrotron radiation. The crystals are stable on exposure to X-rays and suitable for high-resolution X-ray structure determination.

Journal Article↗

PubMed1996

An allel-specific abnormal transcript of the heat shock protein 70 gene in patients with major depression.

Stress-inducible 72-kDa heat shock proteins (HSP70) were encoded on genes in multiple chromosomes. The expression of mRNA transcribed from the gene (HSP70-1) on chromosome 6 was studied using reverse transcript polymerase chain reaction in peripheral blood mononuclear cells of patients with different diseases. The deletion of 29 bp occurred in 5' noncoding and subsequent 133 bp in coding sequences of HSP70 mRNA in patients with major depression (n = 18), while normal subjects (n = 10) and patients with schizophrenia (n = 5), essential hypertension (n = 3), rheumatoid arthritis (n = 7), and Graves' disease (n = 3) had normal mRNA. No such deletion occurred in genomic DNA and no protein was translated from deleted mRNA. The allel-specific abnormal transcript of the HSP70 gene on chromosome 6 thus may underlie the altered stress and/or immune response in major depression.

Alleles↗

PubMed1996

Structural studies of a major hemorrhagin (rhodostoxin) from the venom of Calloselasma rhodostoma (Malayan pit viper).

The complete amino acid sequence, disulfide linkages, glycosylation sites, and carbohydrate structure of rhodostoxin, the major hemorrhagin from Calloselasma rhodostoma (Malayan pit viper), have been determined. This sequence confirmed the deduced amino acid sequence of the putative hemorrhagic protein encoded by the prorhodostomin cDNA of C. Rhodostoma. Rhodostoxin contained four disulfide bonds that link Cys19-Cys60, Cys117-Cys198, Cys157-Cys182, and Cys159-Cys165. It is the first four-disulfide proteinase reported among all known venom metalloproteinases, which are either of the two-disulfide or three-disulfide type. Peptide-mapping and dot-blotting experiments showed the presence of two glycopeptides. Subsequent sequencing of these peptides established that the N-glycosylation sites are located at residues 91 and 181 of the amino acid sequence of the matured protein. Mass spectrometric analyses of these glycopeptides showed that they contain an oligosaccharide structure consisting of 4 units of N-acetylglucosamine, 5 units of hexose, 1 unit of fucose, and 2 units of neuraminic acids. The complete carbohydrate structure was then established by 2-D mapping analysis of the pyridylamino-oligosaccharides after hydrazinolysis and pyridylamination of the glycan chains.

Amino Acid Sequence↗

PubMed1996

Inhibitory effects of alkyl gallate and its derivatives on fatty acid desaturation.

Alkyl gallate, which is known as an antioxidant, intensively inhibited delta 5 and delta 6 desaturation in both rat liver microsomes and an arachidonic acid-producing fungus Mortierella alpina 1S-4. The rat liver microsomal delta 5 and delta 6 desaturases were inhibited by gallic acid esterified with alcohols with various numbers of carbons, suggesting that the necessary structure in an esterified alcohol for the inhibition is not so strict. Among the three hydroxy groups in gallic acid, the m-hydroxy group was shown to be the necessary structure. Kinetic analyses revealed that propyl gallate is a noncompetitive inhibitor of delta 5 desaturase (Ki = 2.6.10(-5)M) and delta 6 desaturase (Ki = 1.7.10(-4) M). These data indicate that alkyl gallate is a new type of desaturase inhibitor and different from known natural inhibitors, i.e., sesamin and curcumin.

Animals↗

PubMed1996

Postnatal development of protein gene product 9.5- and calcitonin gene-related peptide-like immunoreactive nerve fibers in the rat temporomandibular joint.

BACKGROUND: The aim of this study was to investigate the little-known postnatal development of innervation in the temporomandibular joint (TMJ). METHODS: We therefore investigated the protein gene product 9.5-like immunoreactive (PGP-LI) and calcitonin gene-related peptide-like immunoreactive (CGRP-LI) nerves in the rat TMJ from days 0-24 in wholemount preparations and frozen sections by immunohistochemistry using the avidin-biotin-peroxidase complex method. RESULTS: At birth, PGP-LI and CGRP-LI nerves were observed only in the disk attachment but not in the peripheral portion of the disk. From days 3-6, the peptidergic nerves associated with the development of synovial cells were first found in the anterior (3rd day) or posterior (6th day) peripheral portion of the disk in which synovial cells later appear. From days 7-10, incisal occlusion started and simultaneously the distinct networks of the nerves were formed in four peripheral portions of the disk. On the 14th day, the PGP-LI nerves were first more numerous than the CGRP-LI nerves only in the anterior disk attachment. From days 18-24, the occlusion between the first molars started. Simultaneously, the distribution and number of the nerves were comparable to those of adult rats. However, from days 0-24, the central portion of the disk (actual disk) did not contain any nerves. CONCLUSIONS: These findings suggest that functional stimuli such as sucking and mastication might be associated with the development of nerves in the TMJ during the early and late postnatal days, whereas the actual disk except for both disk attachment and peripheral portion of the disk is not innervated and does not contribute to the afferent sensory information from the TMJ. The presence of peptidergic nerves prior to the appearance of synovial lining cells furthermore suggests that the neuropeptide released from these nerves may affect on the differenciation and maturation of synovial cells.

Age Factors↗

PubMed1996

Inhibition by tunicamycin of mucin synthesis, not morphological changes, in epidermis during retinol-induced mucous metaplasia of chick embryonic cultured skin.

BACKGROUND: Our previous studies have shown that epidermal mucous metaplasia of chick embryonic skin can be induced by culture in medium containing 20 microM retinol for only 8 hr and then in a chemically defined medium without retinol for 2 days and that retinol primarily affects the dermal cells, which then transform the epithelial cells into mucus-secreting cells. METHODS: Tarsometatarsal skin of 13-day-old chick embryo was cultured with 20 microM retinol for 1 day and then without the vitamin but with 0.1 microgram/ml tunicamycin for 5 days. Effect of tunicamycin on epidermal mucous metaplasia was studied biochemically and morphologically. RESULTS: Tunicamycin, which prevents the formation of N-glycans and inhibits maturation or morphological organization of various epithelial cells, irreversibly inhibited the synthesis of sulfated glycoproteins (O-glycans, mucin) in the epidermis only when applied to retinol-pretreated skin. Microvilli on the surface of the cells were well developed, but mucous granules surrounded by a limiting membrane were not observed in the upper cell layer of the epidermis, and many vesicles without electron-dense materials (mucin) and dilated rough endoplasmic reticulum were seen in the intermediate cell layers of the epidermis. When recombinants of 13-day-old normal epidermis and cultured dermis, which had been treated with retinol for 24 hr and with only tunicamycin for 2 days, were cultured without the antibiotic for 5 days, epidermal mucous metaplasia was induced. CONCLUSION: These results suggest that tunicamycin did not prevent morphological changes induced by retinol but inhibited mucin synthesis by a direct action on the epidermis of retinol-pretreated skin. Because in some cell-line mucin precursors contain high mannose N-linked oligosaccharides side chains, tunicamycin may have inhibited mucin synthesis. Interaction between epidermal basal cells and retinol-pretreated dermal fibroblasts is prerequisite for epidermal mucous metaplasia. Thus, the present study suggests that N-linked protein glycosylation is not required for this interaction.

Animals↗

PubMed1996

Temperature-dependent increase in the DNA-binding activity of a heat shock factor in an extract of tobacco cultured cells.

The DNA-binding activity of a tobacco heat shock factor (HSF) was induced by heat treatment (37-40 degrees C) of a cell-free extract that contained extra-nuclear fraction, but not in an extract of isolated nuclei. These observations suggest that an inactive form of HSF can directly recognize and transduce the heat shock signal and that such transduction requires components of the extranuclear fraction. Addition of ATP or of most other nucleoside triphosphates reduced the binding of the HSF to the heat shock element (HSE) in the same extract, and removal of ATP by dialysis from the extract restored the ability of the HSF to bind to DNA. The restored activity of the HSF could be eliminated again by a second addition of ATP. Our observations provide the first example of the involvement of ATP in the regulation of the reversible changes in HSF that control its ability to bind to HSEs in a cell-free extract.

Adenosine Triphosphate↗

PubMed1996

Changes of lymphocyte subpopulations in pigs infected with porcine reproductive and respiratory syndrome (PRRS) virus.

Abnormal changes of T-cell subpopulation were observed in the peripheral blood lymphocytes of pigs infected with porcine reproductive and respiratory syndrome (PRRS) virus. Pigs with naturally occurring PRRS revealed increases in CD2+ and CD8+ cells, and decreases in CD4+ cells and the ratios of CD4+/CD8+ cells. Specific-pathogen-free pigs inoculated with PRRS virus showed remarkable decreases in total lymphocytes, CD4+ and CD2+ cells on Postinoculation Day (PID) 3. The decline of CD4+ cells continued for at least 14 days, while CD2+ cells showed a tendency to increase thereafter. On the other hand,CD8+ cells slightly decreased in number on PID 3, and then increased remarkably; their number was significantly larger on PIDs 28 and 35 than on PID 0. The ratios of CD4+/CD8+ cells were significantly low between PIDs 3 and 28 as compared with PID 0. However, there were no differences in thymocyte subpopulations between infected and non-infected pigs, suggesting that the PRRS virus does not modulate intrathymic T-cell differentiation. In an experiment with peripheral blood mononuclear cell cultures, PRRS virus caused neither alteration of T-cell subpopulations nor cell proliferation, suggesting that the virus is not cytotoxic for CD4+ cells and not mitogenic for CD8+ cells.

Animals↗

PubMed1996

Improvement of rat liver graft function by insulin administration to donor.

BACKGROUND & AIMS: The nutritional status of a donor is considered to be an important factor affecting organ viability. The purpose of the present study was to examine hepatic energy status in connection with posttransplantation liver function. METHODS: The following five groups of donor rat livers were prepared and stored in University of Wisconsin solution at 4 degrees C: fasted group, rats fasted for 24 hours; fed group, rats provided standard laboratory chow; fed plus insulin group, fed rats treated with insulin before harvest; fasted plus insulin group, fasted rats treated with insulin before harvest; and fed plus glucagon group, fed rats pretreated with glucagon. RESULTS: Hepatic levels of adenosine triphosphate and total adenine nucleotides were maintained in the decreasing order of the fed plus insulin, fed, fasted plus insulin, fasted, and fed plus glucagon groups during preservation. Lactate production rate and fructose 2,6-bisphosphate level increased in the order of the fed plus insulin, fed, fed plus glucagon, fasted, and fasted plus insulin groups. Liver function after transplantation evaluated by the bile flow rate and enzyme leakage was well restored in the fed plus insulin group. CONCLUSIONS: Insulin administration to nutritionally well-supported livers before harvest improved energy metabolism during preservation and liver function after transplantation.

Adenosine Triphosphate↗

PubMed1996

Targeted disruption of the DNA repair methyltransferase gene renders mice hypersensitive to alkylating agent.

Alkylation of DNA at the O(6)-position of guanine is one of the most critical events leading to induction of mutation as well as to cancer. The enzyme O(6)-methylguanine-DNA methyltransferase repairs this and related lesions in DNA. By means of gene targeting, we established mouse lines deficient in the methyltransferase gene and tissues from these mice contained no methyltransferase activity. Administration of methylnitrosourea to these gene-targeted mice led to early death, and normal mice treated in the same manner showed no untoward effects. In mice given methylnitrosourea treatment, the bone marrow became hypocellular and there was a drastic decrease in the number of leukocytes and platelets, thereby indicating an impaired reproductive capacity of hematopoietic stem cells. Methyltransferase apparently protected these mice from the pancytopenia caused by the alkylating agent.

Alkylating Agents↗

PubMed1996

Kobe earthquake and reduced sperm motility.

We investigated a possible relationship between the Kobe earthquake (January 17, 1995) and the quality of semen. We assessed sperm concentration and motility of 27 male patients who had a concentration of more than 30 million/ml and >40% sperm motility within 5 months before the earthquake. Twelve male patients from districts with a magnitude of <4 on the Richter scale showed no difference in sperm concentration and motility before and after the earthquake. Of 15 male patients from districts with a magnitude of >6, five patients whose houses received no damage showed no distinct changes in sperm concentration and motility. In contrast, 10 patients whose houses were partially or completely destroyed showed significantly (P < 0.001) lower sperm motility after the earthquake than before, although no significant difference of sperm concentration could be observed. Of these latter 10 patients, seven could be followed. In six patients, sperm motility was restored between 2 and 9 months after the earthquake; the sperm motility in one patient, whose father died a victim of the house crash, has not yet recovered. Thus, the acute stress resulting from such a catastrophic earthquake could be a possible cause of reduced sperm motility.

Adult↗

PubMed1996

Metastatic testicular choriocarcinoma of the skin. Report and review of the literature.

Choriocarcinoma is a malignant growth of trophoblastic cells characterized by secretion of human chorionic gonadotropin. Choriocarcinoma usually arises from fetal trophoblasts and rarely arises from germ cells in the testis or ovary or derives from dedifferentiation of other carcinomas. Skin metastasis of choriocarcinoma is rare: only seven cases have been reported in the English and Japanese literature. We report the case of a 22-year-old Japanese man with pure choriocarcinoma of the testis who developed skin metastases that presented as multiple reddish nodules. Microscopic examination of both the primary lesion of the testis and the cutaneous metastasis demonstrated the typical histologic features of pure choriocarcinoma. The patient died 3 months after the initial onset of skin metastasis. Review of the literature indicates that skin metastasis of choriocarcinoma usually occurs as a nodular lesion with the histologically typical feature of the primary disease and signals of poor prognosis.

Adult↗

PubMed1996

Photoallergic drug eruption due to pyridoxine hydrochloride.

Photoallergy to vitamin B6 is very rare; only a few cases of contact dermatitis and one case of photosensitive dermatitis due to pyritinol have been reported. We report here the first case of photoallergic drug eruption due to pyridoxine hydrochloride. A 71-year-old man developed papulo-squamous erythemata which were confined to sun-exposed sites. Photopatch testing, together with the clinical course, was helpful in reaching the initial diagnosis; this was confirmed by an oral challenge test. Photoallergic drug eruption due to vitamin B6 should be considered a rare cause of photosensitive dermatitis.

Aged↗

PubMed1996

Establishment and characterization of high- and low-lung-metastatic cell lines derived from murine colon adenocarcinoma 26 tumor line.

We established and characterized high- (LuM1) and low-lung-metastatic (NM11) cell lines derived from murine colon adenocarcinoma 26 tumor line. LuM1 cell line was established as a clonal cell line from a cultured cell mixture derived from a lung-metastatic nodule after 7 sequential subcutaneous transplantations of lung-metastatic tumors in the abdominal wall of BALB/c mice. NM11 cell line was established from a cultured cell mixture derived from a subcutaneous transplant of murine colon adenocarcinoma 26 tumor cells. LuM1 cells showed marked spontaneous lung metastases, but NM11 cells rarely did. High invasive potential of LuM1 cells was revealed by in vitro invasion assay using Matrigel reconstituted membranes. Rapid retraction was observed in monolayers of human umbilical vein endothelial cells and bovine aortic endothelial cells when LuM1 cells were added on the monolayers. Gelatin zymography and immunochemical examinations with monoclonal antibodies against gelatinase B (Mr 95,000 type IV collagenase) showed secretion of large amounts of the gelatinase by LuM1 cells.

Adenocarcinoma↗