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Biomedical subjects

S Hayashi

Publications and source records attributed to S Hayashi.

At least 163 records · Page 9Linked to original sources
PubMed2000

Activating Ly-49D and inhibitory Ly-49A natural killer cell receptors demonstrate distinct requirements for interaction with H2-D(d).

The activating Ly-49D receptor and the inhibitory Ly-49A receptor mediate opposing effects on natural killer (NK) cell cytotoxicity after interaction with the same major histocompatibility complex ligand, H2-D(d). To compare Ly-49D and Ly-49A interactions with H2-D(d), we created mutations in H2-D(d) and examined the functional ability of these mutants to activate lysis through Ly-49D or to inhibit lysis through Ly-49A. Specific single amino acid changes in either the H2-D(d) alpha(1) helix or the alpha(2) helix abrogated Ly-49D-mediated cytotoxicity, but these changes had no significant effect on Ly-49A-dependent inhibition. Each of three alpha(2) domain mutations in the floor of the peptide binding groove reduced functional recognition by either Ly-49D or Ly-49A, but all three were required to fully abrogate inhibition by Ly-49A. Our studies indicate that Ly-49D/H2-D(d) interactions require distinct determinants compared with Ly-49A/H2-D(d) interactions. These differences have important implications for the integration of activating and inhibitory signals in NK cells.

Amino Acid Substitution↗
PubMed2000

Induction of Th2-directed immune responses by IL-4-transduced dendritic cells in mice.

Dendritic cell (DC)-based vaccines have been used to generate Th1-mediated, protective immunity against cancers and infectious microorganisms. As an attempt to develop a new vaccine protocol for the induction of Th2-directed responses, we introduced an IL-4 plasmid vector into the XS106 DC line (derived from A/J mice). Although relatively small fractions of XS106 cells exhibited apparent intracellular deposition of IL-4, they secreted biologically relevant amounts of the cytokine. IL-4-transduced XS106 DC and control XS106 DC transfected with vector alone were pulsed with KLH and injected s.c. into A/J mice. The overall magnitude of KLH-specific cellular and humoral responses was comparable between the two animal groups. However, they differed in the isotype profile albeit only transiently, with the IL-4-transduced DC group showing higher IgE and lower IgG2a responses, and in the cytokine profile, with spleen cells isolated from the IL-4-transduced DC group producing higher IL-13 and lower IL-12. Thus, delivery of IL-4 gene to relatively small numbers of DC is sufficient to modify the immunological outcome of DC-based vaccines.

Animals↗
PubMed2000

Continuous enzymatic production of peptide precursor in aqueous/organic biphasic medium.

N-(benzyloxycarbonyl)-L-aspartic acid (Z-L-Asp) has generally been used as a carboxyl substrate for the enzymatic synthesis of a precursor of aspartame (synthetic sweetener); however, alternative inexpensive protection groups have been in demand for lowering the total cost of its industrial-scale production. A formyl group (F-) was found to be a more desirable protecting group for the N-terminus of amino acid derivatives due to its low cost of preparation, introduction, and removal. The yield of F-AspPheOMe (N-formyl-L-aspartyl-L-phe- nylalanine methylester), however, was found to be <10% in a conventional aqueous medium. We found that F-L-Asp and L-PheOMe were partitioned mainly to the aqueous phase in an aqueous/organic biphasic medium, whereas F-AspPheOMe partitioned to the organic phase, especially when some extracting agents were added. In this study, simultaneous operation of an enzymatic reaction and a product separation by liquid-liquid extraction was thus applied to the F-AspPheOMe synthesis. We succeeded in synthesizing F-AspPheOMe continuously in an aqueous/tributylphosphate (TBP) biphasic medium with >95% yield, which was about tenfold higher than that in an aqueous monophasic medium.

Organic Chemicals↗
PubMed2000

Estrogen receptor alpha, but not beta, is expressed in the interneurons of the hippocampus in prepubertal rats: an in situ hybridization study.

Estradiol is involved in the differentiation and plasticity of hippocampal neurons. In the CA1 region, estrogen treatment increases dendritic spines and synapse density on pyramidal cells. In the adult hippocampus, immunoreactivity for estrogen receptor alpha (ERalpha) has been reported in inhibitory interneurons, but neither in the pyramidal neurons nor in granule cells. Estrogens also mediate aspects of sexual differentiation of the hippocampus. To examine the possibility that an alteration in expression of the two types of estrogen receptors (ERalpha and ERbeta) in the hippocampus underlies different roles of estrogen and/or ERs during development and in adult life, we applied non-isotopic, digoxigenin (dig)-labeled, in situ hybridization histochemistry (ISHH) for the both ER forms and examined the distribution pattern of their messages in serial, frontal sections over the postnatal period and in the adult. ERalpha mRNA expression was found scattered throughout the hippocampus especially in the hilar region of the dentate gyrus, and in the strata radiatum and pyramidale in the cornus ammonis at postnatal days (PND) 14, 21 and 35. In the hilus of the dorsal hippocampus, the density of ERalpha-labelled cells was greater in the rostro-medial aspect, while less in the lateral and the caudal region. In the ventral hippocampus the signals for ERalpha mRNA were also found in relatively high density in the hilus. No significant sex difference in distribution and intensity of the ERalpha mRNA positive cells were detected. The hippocampal distribution of ERalpha mRNA expression at PND 14 remained the same on PND 21 and 35 and in adulthood. As reported for adults, ERalpha mRNA signals appear to be in interneurons of the hippocampus but neither in the pyramidal cells nor in the dentate granular cells based on their size and location. In contrast to the result of ERalpha, no clear signals for ERbeta mRNA were detected in the hippocampus across all ages examined, whereas they were clearly detected in the hypothalamus.

Amygdala↗
PubMed2000

The development of terminal Schwann cells associated with periodontal Ruffini endings in the rat incisor ligament.

The postnatal development of the terminal Schwann cell, an analogue of the lamellar cell in cutaneous sensory receptors, was examined by histochemistry for non-specific cholinesterase and immunohistochemistry for S-100 protein in the periodontal Ruffini endings of the rat incisor. Double immunohistochemistry for S-100 protein and protein gene product 9.5 (PGP 9.5) was also performed to examine the relationship between terminal Schwann cells and axons. Histochemistry for non-specific cholinesterase was able to demonstrate the age-related development of the terminal Schwann cells; the morphology and distribution of the developing terminal Schwann cells became almost identical to those in adults during postnatal days 15-18. Axons showing PGP 9.5-like immunoreactivity elongated and expanded after arrangement of terminal Schwann cells in the alveolus-related part. This suggests that the terminal Schwann cell is important in the development and maturation of the periodontal Ruffini endings.

Animals↗
PubMed2000

Transcriptional activation of urokinase by the Krüppel-like factor Zf9/COPEB activates latent TGF-beta1 in vascular endothelial cells.

Understanding the regulation of genes controlling fibrinolysis and matrix homeostasis is essential for elucidating the basis of tissue repair. A recently described novel Krüppel-like factor, Zf9, is up-regulated in acute liver injury in activated hepatic stellate cells. Because Zf9 can be induced widely, its activity was examined in vascular endothelium, a key cell in vascular injury. Zf9 is induced as an immediate-early response gene in bovine aortic endothelial cells (BAECs) following treatment with serum or phorbol ester. Zf9 transcriptionally activates urokinase plasminogen activator (uPA). Recombinant Zf9-GST binds to wild-type but not mutated 'GC-box' motifs within the human uPA promoter (-63 to -32), with greatest affinity to the middle of 3 contiguous GC boxes. Transient transfection of Zf9 drives transactivation of a full-length uPA promoter- and GC box-construct, but not a uPA promoter-construct devoid of GC boxes. Transactivation of uPA by Zf9 is also supported in Drosophila S2 cells. Most importantly, transiently transfected Zf9 up-regulates endogenous uPA messenger RNA and activity in BAECs, resulting in increased bioactive transforming growth factor-beta (TGF-beta) via enhancement of proteolytic activation of the latent molecule. Furthermore, concomitant expression of Zf9 and uPA proteins was observed in arterial endothelial cells after balloon injury in rats, suggesting a potential role of Zf9 in uPA expression not only in vitro but also in vivo. These findings suggest a role of Zf9 in the injury response by enhancing uPA synthesis and subsequent activation of latent TGF-beta. (Blood. 2000;95:1309-1316)

Animals↗
PubMed2000

Degradation of ornithine decarboxylase by the 26S proteasome.

Ornithine decarboxylase (ODC) is a key enzyme in polyamine biosynthesis. Turnover of ODC is extremely rapid and highly regulated, and is accelerated when polyamine levels increase. Polyamine-stimulated ODC degradation is mediated by association with antizyme (AZ), an ODC inhibitory protein induced by polyamines. ODC, in association with AZ, is degraded by the 26S proteasome in an ATP-dependent, but ubiquitin-independent, manner. The 26S proteasome irreversibly inactivates ODC prior to its degradation. The inactivation, possibly due to unfolding, is coupled to sequestration of ODC within the 26S proteasome. This process requires AZ and ATP, but not proteolytic activity of the 26S proteasome. The carboxyl-terminal region of ODC presumably exposed by interaction with AZ plays a critical role for being trapped by the 26S proteasome. Thus, the degradation pathway of ODC proceeds as a sequence of multiple distinct processes, including recognition, sequestration, unfolding, translocation, and ultimate degradation mediated by the 26S proteasome.

Animals↗
PubMed2000

Two zebrafish (Danio rerio) antizymes with different expression and activities.

Cellular polyamines are regulated by a unique feedback mechanism involving ornithine decarboxylase (ODC) antizyme. The synthesis of mammalian antizyme requires a programmed translational frameshift event induced by polyamines. Antizyme represses ODC, a key enzyme for polyamine synthesis, through accelerating enzyme degradation by the 26 S proteasome. Antizyme also inhibits the cellular uptake of polyamines. In the present study we isolated two distinct zebrafish (Danio rerio) antizyme cDNA clones (AZS and AZL) from an embryonic library. Their sequences revealed that both clones required translational frameshifting for expression. Taking account of +1 frameshifting, AZS and AZL products were 214 and 218 residues long respectively and shared 51.8% amino acid identity. In rabbit reticulocyte lysates, both mRNA species were translated through spermidine-induced frameshifting. The presence of the two antizyme mRNA species in embryos, adult fish and a cultured cell line was confirmed by Northern blot analysis. The ratio of AZS mRNA to AZL mRNA in the adult fish was 1.8-fold higher than in the embryos. Whole-mount hybridization in situ demonstrated that both mRNA species are expressed in every tissue in embryo, but predominantly in the central nervous system and the eyes. Bacterial expression products of both cDNA species inhibited ODC activity, but only the AZS product accelerated ODC degradation in vitro. These results show that both zebrafish antizymes are induced by polyamines but their mRNA species are expressed differently during development. The difference in activities on ODC degradation suggests their functional divergence.

Amino Acid Sequence↗
PubMed2000

Influence of preparation conditions on the microstructure and bioactivity of alpha-CaSiO(3) ceramics: formation of hydroxyapatite in simulated body fluid.

Two different reagents, NaOH and NH(4)OH, were used to precipitate CaSiO(3) precursor powders from ethanol solutions of Ca(NO(3))(2). 4H(2)O and Si(OC(2)H(5))(4). The resultant powders of different Ca/Si ratio and residual Na(2)O content exhibited significant differences in the microtexture of the resulting sintered alpha-CaSiO(3) ceramics. The microtexture of the ceramics from the NaOH system (CS-Na) contained smaller grain sizes and a thicker glassy phase at the grain boundaries than those produced using NH(4)OH (CS-NH). The CS-Na ceramics were soaked in a simulated body fluid (SBF) at 36.5 degrees C for 2 h and 1, 5, 6, 10, 21, and 30 days while the CS-NH ceramics were soaked for 1, 5, 7, 15, 20, and 25 days using the same conditions. Hydroxyapatite (HAp) formed on the surfaces of both samples but at different formation rates due to differences in the microstructure. The CS-Na ceramics showed faster HAp formation because their smaller alpha-CaSiO(3) grains dissolved more readily, allowing the calcium concentration in the SBF quickly to approach the appropriate condition for nucleation of HAp. In addition, the thicker glassy phase at the grain boundaries facilitated a faster formation of silanol on the surface of the amorphous SiO(2) interlayer, a reaction that is considered to be a prerequisite for HAp formation. The formation of the HAp layer on the CS-Na ceramics therefore was very fast (12 microm/day), and their surfaces were covered completely within 5 days. A layer thickness of about 110 microm was achieved in 30 days, in contrast with the CS-NH ceramics, which took about 25 days to be fully covered with a 60-microm layer of HAp.

Body Fluids↗
PubMed2000

An autopsy case of autosomal-recessive juvenile parkinsonism with a homozygous exon 4 deletion in the parkin gene.

We report the neuropathologic and genetic features of a 70-year-old man with autosomal-recessive juvenile parkinsonism (AR-JP). At the age of 32 years, he developed a dystonic gait, followed by hand tremor, rigidity, bradykinesia, and impaired postural reflex. Levodopa was effective in ameliorating these symptoms. Pathologic examination of autopsy specimens from this patient revealed loss of pigmented neurons and gliosis in the substantia nigra pars compacta (SNPC), being most pronounced in the medial and ventrolateral regions, and locus ceruleus (LC). The melanin content of the remaining SNPC neurons was low. This feature was less marked in the LC. There were no Lewy bodies, as confirmed by immunostaining for alpha-synuclein. An additional, significant finding in this patient was neuronal loss and fibrillary gliosis in the substantia nigra pars reticulata; this feature has not been reported previously in AR-JP. Gene analysis revealed that this autopsied patient and his siblings had the parkin gene mutation (homozygous exon 4 deletion) that is responsible for the disease.

Aged↗
PubMed2000

Development of astroglial elements in the suprachiasmatic nucleus of the rat: with special reference to the involvement of the optic nerve.

The development of astroglial cells and the effect of the retinohypothalamic tract on it were studied by vimentin and glial fibrillary acidic protein (GFAP) immunocytochemistry in the suprachiasmatic nucleus (SCN) of the rat. At the embryonic stage, vimentin-immunoreactive (VIM-IR) radial glia, precursors of astrocytes, were dominant. However, their filaments vanished in the first few postnatal days. Instead of VIM-IR glial filaments, GFAP-immunoreactive (GFAP-IR) astrocytes appeared at E20 and grew rapidly from the P3 stage. GFAP immunoreactivity in the ventrolateral portion of the SCN (VLSCN) was measured using a computer-assisted image analyzing system. In normal rats, GFAP immunoreactivity showed a stepwise pattern with two slopes at P3-P4 and P20-P25. Bilaterally eye-enucleated rats operated on the day of birth showed lower GFAP immunoreactivity than normal rats and the GFAP immunoreactivity did not increase between P20 and P25 when GFAP-IR glial processes rapidly expand. Electron microscopic investigation at P50 (adult stage) revealed that neurons in the VLSCN had often direct apposition without astroglial processes and the frequency of this finding was significantly higher in eye-enucleated rats than in the control rats. These findings strongly suggest that the postnatal development of astroglial elements, particularly the expansion of GFAP-IR processes in the SCN, is regulated by retinohypothalamic projection.

Afferent Pathways↗
PubMed2000

27Al NMR study in ZrNiAl.

We have studied the microscopic properties of the hexagonal ZrNiAl, a model compound for a wide family of intermetallic compounds crystallizing in this type of structure, by using 27Al NMR spectroscopy. We have investigated the lineshape of static and MAS NMR spectra as a function of magnetic field strength (4.7-9.4 T) and temperature (5-300 K). Our data indicate that the 27Al NMR spectra result from a combined effect of quadrupole and anisotropic shift interactions. The 27Al nuclei are in an environment characterized by the quadrupole coupling constant e2qQ/h of 3.3 MHz, asymmetry parameter etaQ of 0.42, isotropic shift delta(iso) of 393 ppm, shift anisotropy delta(anis) = delta(zz) - (delta(xx) + delta(yy))/2 of 150 ppm, and asymmetry factor etaS of 0.5. They are found to be temperature independent. The spin-lattice relaxation rate measured at 7.05 T is proportional to the temperature with T1T = 135 s K. The mechanisms responsible for observed values of delta(iso), delta(anis), T1T, and the enhanced Korringa constant are discussed.

Aluminum↗
PubMed2000

Role of Pararectal Retroperitoneal Approach in Abdominal Aortic Aneurysm Repair.

To evaluate the appropriate use of the pararectal retroperitoneal approach (RPA) in abdominal aortic aneurysm (AAA) repair, we retrospectively compared the RPA with the transperitoneal approach (TPA). Fifty-four patients in whom the RPA was used and 92 who had the TPA were included in the study. The mean operating time was 286 minutes with the RPA and 252 minutes with the TPA. Intraoperative blood transfusions were necessary in 9.3% of patients who had the RPA and 16.2% who had the TPA. As assessed by computerized tomographic scanning, the mean size of the postoperative retroperitoneal hematoma was 440 mL in the RPA group and 180 mL in the TPA group. Ileus resolved a mean of 3 days postoperatively in the RPA group and 5 days postoperatively in the TPA group. The mean length of postoperative hospitalization was 21 days in the RPA group and 23 days in the TPA group. Prolonged postoperative ileus occurred in no patients in the RPA group and in three patients in the TPA group. None of the differences in outcomes between the groups were significant. Our findings indicate that, in carefully selected patients, the pararectal RPA is suitable for AAA repair.

Journal Article↗
PubMed2000

Cerebral protection selection in aortic arch surgery for patients with preoperative complications of cerebrovascular disease.

OBJECTIVE: Retrograde perfusion is gaining acceptance as a means of cerebral protection, but it remains unclear how long the brain is protected and whether it is effective in patients with preoperative cerebrovascular disease. METHODS: From January 1989 to August 1999, 205 patients--118 male and 87 female patients who ranged 12 to 86 years old, mean: 65.5 years old--underwent surgery at our hospital for aortic arch aneurysm using cerebral protection. We focused on mortality, stroke incidence and perioperative risk factor between 2 groups--selective cerebral and retrograde cerebral perfusion--also studying patients with preoperative cerebrovascular disease that influenced postoperative stroke. RESULTS: The hospital mortality was 11.7% (selective cerebral perfusion group: 12%, retrograde group: 10.9%). Stroke occurred in 11 patients (5.3%), 4.7% in the selective cerebral perfusion group and 7.3% in the retrograde group. Preoperative cerebrovascular disease does not appear to be a risk factor for postoperative brain damage in aortic arch surgery. Regarding total replacement of the aortic arch, the incidence of postoperative brain damage in the retrograde group with preoperative cerebrovascular disease was higher than that in another group (p = 0.072). Cardiopulmonary bypass time and selective cerebral perfusion time in the patients with postoperative stroke were significantly longer than that in non-stroke group. CONCLUSIONS: Preoperative cerebrovascular disease did not appear to be a risk factor in postoperative neurological deficit in the selective cerebral perfusion group. Prolonged selective cerebral perfusion time and cardiopulmonary bypass time may, however, lead to brain edema and cause neurological deficit.

Adolescent↗
PubMed2000

NACP/alpha-synuclein-positive filamentous inclusions in astrocytes and oligodendrocytes of Parkinson's disease brains.

The precursor of the non-Abeta component of Alzheimer's disease amyloid (NACP), also called alpha-synuclein, is a major component of Lewy bodies in Parkinson's disease (PD) as well as of neuronal and oligodendroglial cytoplasmic inclusions in multiple system atrophy. We previously reported argyrophilic, tau-negative glial inclusions in the midbrains of patients with PD and have now conducted immunocytochemical and ultrastructural examinations. The PD glial inclusions also are immunoreactive for NACP/alpha-synuclein, but not for beta-synuclein, and ultrastructurally are composed of filamentous structures about 25-40 nm in diameter. Double immunolabeling showed that the inclusions were present in both astrocytic and oligodendroglial cells. They were located within the substantia nigra in 13 of 30 patients with PD and outside the nigra in 24. The number of inclusions was correlated with the severity of nigral neuronal loss. These findings indicate that abnormal accumulation of NACP/alpha-synuclein in glial cells is a pathological feature of PD related to its progression.

Aged↗
PubMed2000

Ganglioglioma with a tanycytic ependymoma as the glial component.

We studied a cystic ganglioglioma (GG) located in the right frontal lobe of the brain. Interestingly, the fibrillary spindle glial cells were often arranged in a fascicular pattern, and the generally uniform, round-to-oval delicate nuclei appeared to resemble those of ependymoma; and the neoplastic neurons often contained neurofibrillary tangles (NFTs). The glial component was positive for glial fibrillary acidic protein and occasionally contained granular or microvesicular structures positive for epithelial membrane antigen. Ultrastructural investigation revealed that the glial cells were ependymal in nature; intracytoplasmic lumina and intercellular microrosettes lined with cilia and microvilli, as well as long zonulae adherentes, were evident. In addition, chromogranin A-positive granular staining, neurosecretory-granule-like structures, and parallel arrays of microtubules were sometimes associated with the blood vessels. We considered the present case to be an unusual example of GG with an ependymoma, more precisely a tanycytic ependymoma, as the glial component; to our knowledge, the existence of ependymoma as the main glial component of this particular tumor has not been described before. The occurrence of NFTs, which has been reported in several cases of GG, was an additional, unusual feature.

Brain Neoplasms↗