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S Miyata

Publications and source records attributed to S Miyata.

At least 19 recordsLinked to original sources
PubMed2006

Clinicopathological features of pyothorax-associated lymphoma; a retrospective survey involving 98 patients.

To investigate clinicopathological features of pyothorax-associated lymphoma (PAL), we examined medical records of 98 patients (88 males and 10 females) with PAL at a median age of 70 years (range 51-86). Seventy-nine patients had a history of artificial pneumothorax. Median interval between diagnosis and artificial pneumothorax was 43 years (range 19-64). At diagnosis, performance status (PS) was 0-1 (n=56) and 2-4 (n=42). Clinical stages were I (n=42), II (n=26), III (n=8) and IV (n=22). Pathological diagnosis comprised diffuse large-B-cell (n=78) and peripheral T-cell lymphoma (n=1). Seventeen were treated supportively. The other 81 received aggressive treatments; chemotherapy (n=52), radiotherapy (n=7), surgery (n=4) and combination (n=18). Five-year overall survival (OS) was 0.35 (95% confidence interval, 24% to 45%). Causes of deaths were PAL (n=39), respiratory failure (n=13) and others (n=12). Multivariate analysis identified prognostic factors for OS; lactate dehydrogenase levels [hazard ratio (HR)=2.36; P=0.013], sex (female versus male) (HR=0.15; P=0.01), PS (2-4 versus 0-1) (HR=2.20; P=0.02), clinical stages (III/IV versus I/II) (HR=1.95; P=0.037) and chemotherapy (HR=0.31; P=0.01). Most patients with PAL are elderly and have comorbidities, while some of them achieve durable remission with appropriate treatments. These findings prompt us to establish an optimal treatment strategy on the basis of risk stratification of individual patients.

Aged↗
PubMed2006

Immunohistochemical and western analyses of protein arginine N-methyltransferase 3 in the mouse brain.

The distribution of protein arginine N-methyltransferase 3 (PRMT3) was investigated in the mouse brain using indirect immunofluorescence. PRMT3 was observed to be localized in the cell bodies and dendrites of neurons but not in the axons and glial cells, indicating that PRMT3 is involved in neuronal function. The distribution of the immunoreactive neurons in the brain was uneven, indicating that PRMT3 plays a role in specific neuronal systems such as the motor and limbic systems, as well as functions related to the cerebellum. The present ontogenetic analysis of PRMT1 and PRMT3 using Western blot methodology clearly revealed that PRMT3 develops during the perinatal stage and its expression is maintained even in adulthood. PRMT1, on the other hand, is expressed transiently during the early embryonic stage. These findings indicate that PRMT3 is related with neuronal function in both young and adult brains, while PRMT1 has roles in the immature brain, such as the formation of neural circuits.

Age Factors↗
PubMed2006

Effect of manipulating serum phosphorus with phosphate binder on circulating PTH and FGF23 in renal failure rats.

Phosphorus directly controls parathyroid hormone (PTH) synthesis and secretion. Serum levels of the novel phosphate-regulating hormone, fibroblast growth factor 23 (FGF23), are positively correlated with hyperphosphatemia in patients with chronic renal insufficiency (CRI). We proposed that changes in serum PTH and FGF23 levels might be associated with changes in serum phosphorus levels caused by the phosphate binder sevelamer hydrochloride (sevelamer, i.e. crosslinked poly[allylamine hydrochloride]). Rats were fed a diet containing adenine for 4 weeks to establish CRI. Animals were then offered either a normal diet or a diet containing 1 or 3% sevelamer for 8 weeks continuously, or intermittently with sevelamer diet or a normal diet offered for alternating 2-week periods. Changes in the serum levels of phosphorus, calcium, PTH, FGF23, and 1alpha,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3)) were monitored over time. Adenine-treated rats developed severe CRI, with markedly elevated serum levels of phosphorus, PTH and FGF23, and reduced levels of serum 1,25(OH)(2)D(3). Continuous treatment with sevelamer suppressed these increases throughout the study period. Serum phosphorus, PTH, and FGF23 levels decreased rapidly when sevelamer treatments commenced and recovered rapidly once they were discontinued. However, the changes in serum FGF23 levels began after the onset of changes in serum phosphorus and PTH levels. In conclusion, circulating PTH, and FGF23 levels can be promptly manipulated through the control of serum phosphorus levels. Moreover, phosphate-binder treatment can effectively inhibit the elevation of serum FGF23 levels, as well as PTH levels, under conditions of CRI.

Adenine↗
PubMed2005

Construction of perineuronal net-like structure by cortical neurons in culture.

Perineuronal nets consisting of chondroitin sulfate proteoglycans and hyaluronic acid are associated with distinct neuronal populations in mammalian brain. Whether neurons or glia cells produce these surface-associated chondroitin sulfate proteoglycan perineuronal nets has remained in question. In the present study, we observed perineuronal net-like structure by rat cortical neurons in dissociated culture using Wisteria floribunda aggulutinin, hyaluronic acid binding protein, and the antibodies recognizing chondroitin sulfate proteoglycans. The double labeling experiments showed that perineuronal net-like structure labeled with Wisteria floribunda aggulutinin was observed often at parvalbumin-positive neurons in dissociated cortical culture without glia. Perineuronal net-like structure was not seen at the early stage of culture, but they became visible concomitantly with neuronal maturation after longer culture. High magnification observation further demonstrated that Wisteria floribunda aggulutinin labeling on cortical neurons was seen as numerous puncta along surface of somata and proximal dendrites, but not axons and synapses. Perineuronal net-like structure on cultured neurons was also visualized using chondroitin sulfate proteoglycan-specific antibodies and hyaluronic acid binding protein. Double labeling study demonstrated that perineuronal net-like structure in cultured cortical neurons was composed of chondroitin sulfate proteoglycans such as neurocan and phosphacan. The hyaluronidase treatment of live neurons abolished cellular labeling of hyaluronic acid binding protein and concomitantly diminished that of Wisteria floribunda aggulutinin. These results indicate that cultured cortical neurons are able to construct perineuronal net-like structure without glial cells.

Animals↗
PubMed2005

A review of the action of tacrolimus (FK506) on experimental models of rheumatoid arthritis.

Tacrolimus (FK506) is an immunosuppressive drug, widely used for organ transplantation and atopic dermatitis. Tacrolimus exerts its immunosuppressive effects primarily by interfering with the activation of T cells, via inhibition of calcineurin. Recent clinical studies have also demonstrated the efficacy of tacrolimus in the treatment of rheumatoid arthritis (RA), an autoimmune disease in which T cells play a pivotal role in pathogenesis. Inflammatory cytokines such as TNF-alpha, IL-1 beta, and IL-6 are involved in development of the disease. Recently, modes of action of tacrolimus on RA have been intensively studied in in vitro and animal arthritis models, demonstrating that tacrolimus exerts various novel actions as an anti-rheumatic drug. The pharmacological action of tacrolimus suggests that it has potential to specifically suppress the production of pathogenic inflammatory cytokines with a low frequency of infection, improve joint inflammation and bone/cartilage destruction, fully recover loss of functional status, exert rapid relief in arthritic pain, and promote osteogenic and chondrogenic differentiation. Here we review the action of tacrolimus on experimental models of RA, with a focus on our recent studies, and provide further insight into experimental models used for identifying efficacious anti-rheumatic drugs.

Analgesics↗
PubMed2005

Neuronal expression of the chondroitin sulfate proteoglycans receptor-type protein-tyrosine phosphatase beta and phosphacan.

Receptor-type protein-tyrosine phosphatase beta (RPTPbeta) and its spliced variant phosphacan are major components of chondroitin sulfate proteoglycans in the CNS. In this study, expression and localization of RPTPbeta and phosphacan were examined in developing neurons by immunological analyses using 6B4, 3F8, and anti-PTP antibodies and reverse transcription-polymerase chain reaction (RT-PCR). Light microscopic immunohistochemistry showed that 6B4 RPTPbeta/phosphacan immunoreactivity was observed around neurons in the cortical plate. Further ultrastructural observation showed that 6B4 RPTPbeta/phosphacan immunoreactivity was observed mainly at the membrane of migrating neurons and radial glia. Immunocytochemical analysis revealed that RPTPbeta immunoreactivity was observed in cultured cerebral, hippocampal, and cerebellar neurons in addition to type-1 and type-2 astrocytes. Western analysis further demonstrated that the shorter receptor form of RPTPbeta (sRPTPbeta) was detected from cell lysate of cortical and hippocampal neurons using 6B4 and anti-PTP antibodies, while sRPTPbeta of cerebellar neurons and type-1 astrocytes was recognized only by anti-PTP antibody. Phosphacan was detected from neuronal culture supernatants of cortical, hippocampal, and cerebellar neurons, but not from type-1 astrocytes using 6B4 and 3F8 antibodies. RT-PCR analysis demonstrated the prominent expression of sRPTPbeta and phosphacan mRNAs in cortical neurons, and that of sRPTPbeta mRNA in type-1 astrocytes. During culture development of cortical neurons, the immunoreactivity of 6B4 sRPTPbeta was observed entirely on the neuronal surface including somata, dendrites, axons, and growth cones at earlier stages of cortical neuronal culture such as stages 2 and 3, while, after longer culture, 6B4 sRPTPbeta immunoreactivity in stages 4 and 5 neurons was detected at dendrites and somata and disappeared from axons, and was not observed over axonal terminals and postsynaptic spines. These results demonstrate that neurons are able to express sRPTPbeta on their cellular surface and to secrete phosphacan, and neuronal expression of sRPTPbeta may modulate neuronal differentiation including neuritogenesis and synaptogenesis.

Animals↗
PubMed2005

Impaired platelet function in a patient with P2Y12 deficiency caused by a mutation in the translation initiation codon.

In this study, we have identified a patient (OSP-1) with a congenital P2Y12 deficiency showing a mild bleeding tendency from her childhood and examined the role of P2Y12 in platelet function. At low concentrations of agonists OSP-1 platelets showed an impaired aggregation to several kinds of stimuli, whereas at high concentrations they showed a specifically impaired platelet aggregation to adenosine diphosphate (ADP). ADP normally induced platelet shape change and failed to inhibit PGE1-stimulated cAMP accumulation in OSP-1 platelets. Molecular genetic analysis revealed that OSP-1 was a homozygous for a mutation in the translation initiation codon (ATG to AGG) in the P2Y12 gene. Heterologous cell expression of wild-type or mutant P2Y12 confirmed that the mutation was responsible for the deficiency in P2Y12. OSP-1 platelets showed a markedly impaired platelet spreading onto immobilized fibrinogen. Real-time observations of thrombogenesis under a high shear rate (2000 s(-1)) revealed that thrombi over collagen were small and loosely packed and most of the aggregates were unable to resist against high shear stress in OSP-1. Our data suggest that secretion of endogenous ADP and subsequent P2Y12-mediated signaling are critical for platelet aggregation, platelet spreading, and as a consequence, for stabilization of thrombus.

Aged↗
PubMed2004

Genotyping of single nucleotide polymorphism using model-based clustering.

MOTIVATION: Single nucleotide polymorphisms have been investigated as biological markers and the representative high-throughput genotyping method is a combination of the Invader assay and a statistical clustering method. A typical statistical clustering method is the k-means method, but it often fails because of the lack of flexibility. An alternative fast and reliable method is therefore desirable. RESULTS: This paper proposes a model-based clustering method using a normal mixture model and a well-conceived penalized likelihood. The proposed method can judge unclear genotypings to be re-examined and also work well even when the number of clusters is unknown. Some results are illustrated and then satisfactory genotypings are shown. Even when the conventional maximum likelihood method and the typical k-means clustering method failed, the proposed method succeeded.

Algorithms↗
PubMed2004

Inflammatory cytokine levels in paw tissues during development of rat collagen-induced arthritis: effect of FK506, an inhibitor of T cell activation.

OBJECTIVE AND DESIGN: To characterize rat collagen-induced arthritis (CIA) on the basis of levels of inflammatory cytokines, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6 in paw tissues, and further investigate the effect of FK506 (tacrolimus), a potent inhibitor of T cell activation, on cytokine levels. METHODS: CIA was induced in female Lewis rats. The volume of hindpaws was measured before and after collagen immunization. TNF-alpha, IL-1beta and IL-6 levels in paw tissue extracts were determined by ELISA. Proteoglycan contents of cartilage in femoral heads was measured as an indication of cartilage destruction. To assess the effect of FK506 on inflammatory cytokine levels, rats were orally treated with 5 mg/kg of FK506 from days 14-21. RESULTS: TNF-alpha a level in paw tissues did not significantly change compared to levels found before collagen immunization, throughout development of CIA. In contrast, IL-1beta and IL-6 levels in paw tissues significantly increased between day 14 and day 28 after collagen imuninization, when the arthritis was at a developed stage. Therapeutic treatment with FK506 reduced the elevated level of IL-6, but not IL-1beta, in paw tissue. FK506 treatment was effective in suppressing paw swelling and also recovering the loss of proteoglycan contents in the cartilage. CONCLUSIONS: Levels of IL-1beta and IL-6, but not TNF-alpha , in paw tissue were upregulated in association with the development of arthritis in rat CIA. These results suggest that IL-1beta and IL-6, rather than TNF-alpha , may play important roles at local inflammatory sites in producing joint destruction in rat CIA. FK506 may improve arthritis in established stages of CIA, by reducing the elevated level of IL-6.

Animals↗
PubMed2004

Comparison of anti-arthritic properties of leflunomide with methotrexate and FK506: effect on T cell activation-induced inflammatory cytokine production in vitro and rat adjuvant-induced arthritis.

OBJECTIVE AND DESIGN: To examine the effect of leflunomide (LEF) on T cell activation-induced inflammatory cytokine production in human peripheral blood mononuclear cells (PBMC) and rat established adjuvant-induced arthritis (AIA), and compare these effects with methotrexate (MTX) and FK506 (tacrolimus), focusing on improvement of joint function in AIA. METHODS: Human PBMC were cultured with immobilized anti-CD3/CD28 monoclonal antibody to produce tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6. The active metabolite of LEF was used in in vitro study. AIA was induced in female Lewis rats. Paw swelling and grip strength were measured as indicators of arthritis and joint function, respectively. Rats were therapeutically administered LEF (3.2-32 mg/kg) from days 15-24 by oral administration. RESULTS: LEF inhibited anti-CD3/CD28 induced production of TNF-alpha, IL-1beta and IL-6, with IC50 values of 27, 21 and 21 microg/ml, respectively. LEF also suppressed mouse bone marrow cell MTT conversion, with an IC50 value of 15 microg/ml. LEF significantly inhibited paw swelling and loss of grip strength in established AIA at 10 and 32 mg/kg. The inhibition of paw swelling and grip strength loss by LEF was more potent than MTX. However, maximum recovery of grip strength loss by LEF (23.5%) was less potent compared to that with FK506 (57.8%). CONCLUSIONS: LEF inhibited anti-CD3/CD28 induced inflammatory cytokine production in human PBMC at concentrations showing deleterious effects on bone marrow cell proliferation. LEF is superior to MXT in improving arthritis and joint function in established AIA, but is inferior to FK506 in recovering joint function, probably due to its anti-proliferative actions.

Animals↗
PubMed2003

Anti-arthritic properties of FK506 on collagen-induced arthritis in rats.

OBJECTIVE AND DESIGN: To determine the effect of FK506 (tacrolimus) on paw inflammation, TNF-alpha expression in joint, and bone and cartilage destruction in type II collagen-induced arthritis (CIA) model in rats. METHODS: CIA was induced by immunization of female Lewis rats with an emulsion of bovine type II collagen and incomplete Freund's adjuvant. Paw inflammation was assessed by the increase in paw volume. Tumor necrosis factor (TNF) -alpha expression in hind knee joint was assessed by immunohistochemical analysis. Lesions of bone and cartilage were assessed on the basis of histological change in knee joint, radiographic analysis in hind paw, bone mineral density in femora and proteoglycan contents in the cartilage of femoral heads. FK506 at doses of 1, 1.8 and 3.2 mg/kg or its placebo formulation was orally administered to rats for 28 days from the day after immunization (n = 10). Effect of FK506 was compared with that of vehicle (distilled water). RESULTS: FK506 at a dose of 1.8 mg/kg significantly suppressed paw swelling (p < 0.01) and histological change in knee joint (p < 0.05). Tumor necrosis factor (TNF)-alpha was mainly expressed in the region with a marked infiltration of inflammatory cells in the hind knee joint. FK506 (3.2 mg/kg) markedly reduced TNF-alpha expression. FK506 at a dose of 1.8 mg/kg suppressed radiographic changes in hind paw (p < 0.05) and also recovered the decrease in bone mineral density in the femora (p < 0.05). Proteoglycan contents in the cartilage of femoral heads were determined to evaluate the cartilage destruction more quantitatively and found to significantly decrease in CIA rats. FK506 at a dose of 1.8 mg/kg recovered the loss of proteoglycan contents (p < 0.01). CONCLUSION: These results show that FK506 is effective in suppressing inflammation, TNF-alpha expression in joint, and damage to bone and cartilage in rat CIA, and may be useful in the treatment of rheumatoid arthritis.

Animals↗
PubMed2003

Biochemical and ultrastructural analyses of IgLON cell adhesion molecules, Kilon and OBCAM in the rat brain.

Kilon (kindred of IgLON) and opioid-binding cell adhesion molecule belong to the IgLON subgroup of immunoglobulin superfamily together with the limbic system-associated membrane protein and neurotrimin. In the present study, we have analyzed biochemical and ultrastructural characterization of Kilon and opioid-binding cell adhesion molecule such as regional and developmental expression patterns, light and electron microscopic localization, and intermolecular interactions. Western blotting revealed a widespread distribution pattern of Kilon with high expression levels in the olfactory bulb, cerebral cortex, diencephalon, hippocampus, and cerebellum and low expression levels in the medulla oblongata and spinal cord. In contrast, opioid-binding cell adhesion molecule showed a regionally restricted expression pattern with high levels only in the cerebral cortex and hippocampus. Expression of Kilon and opioid-binding cell adhesion molecule was increased gradually during postnatal development and maintained until adulthood. Light microscopic immunohistochemistry demonstrated that the localization of opioid-binding cell adhesion molecule and Kilon coincided well with that of vesicle-associated membrane protein 2, a synaptic marker protein, in the cerebral cortex and hippocampus of adult brain. In the cerebellum, Kilon-immunoreactive puncta were observed to colocalize well with that of vesicle-associated membrane protein 2, while opioid-binding cell adhesion molecule immunoreactivity was observed only at part of synaptic glomeruli in the granular layer and rare in the molecular layer. Electron microscopic analysis revealed that Kilon and opioid-binding cell adhesion molecule immunoreactivity was observed mainly at postsynaptic sites of dendritic and somatic synapses in adult cerebral cortex and hippocampus. Only trace levels of Kilon and opioid-binding cell adhesion molecule were detected in the soluble fraction of a cortical homogenate, although a substantial amount of F3 was present in the soluble fraction. A binding analysis using a cross-linker and the immunoprecipitation technique demonstrated that Kilon and opioid-binding cell adhesion molecule interacted heterophilically and homophilically. These findings show that Kilon and opioid-binding cell adhesion molecule are clearly distinguishable from each other in regional expression and localization, and binding patterns. These differences possibly represent diverse functions of each IgLON molecule.

Animals↗
PubMed2003

Dendrite-associated opioid-binding cell adhesion molecule localizes at neurosecretory granules in the hypothalamic magnocellular neurons.

Opioid-binding cell adhesion molecule (OBCAM) is a member of the immunoglobulin superfamily containing limbic system-associated membrane protein (IgLON) subgroup of glycosylphosphatidylinositol-anchored immunoglobulin cell adhesion molecules. We have previously found that OBCAM is localized preferentially to dendrites compared with somata and terminals of hypothalamic vasopressin-secreting magnocellular neurons. This localization indicates that OBCAM is one of the dendrite-associated cell adhesion molecules. In the present study, we further characterized the localization and the sorting mechanism, and activity-dependent changes of this molecule in vasopressin-secreting magnocellular dendrites. Confocal microscopic observation revealed the preferential localization of OBCAM at the neurosecretory granules in the vasopressin-positive dendrites. Electron microscopic observation using chromogen-intensified and gold-conjugated methods also demonstrated the OBCAM labeling at most of the neurosecretory granules within the dendrites, while the labeling within the somata was observed at only a few neurosecretory granules. I.c.v. colchicine administration resulted in the disappearance of OBCAM immunoreactivity from the dendrites and in its concomitant accumulation at the somata, suggesting that OBCAM is synthesized at the somata and transported to the dendrites by dendrite-associated neurosecretory granules. During the postnatal development, OBCAM immunoreactivity targeted to vasopressin-positive dendrites became clear from at least 3 weeks after birth, although it appeared at only a few somata 2 weeks after birth. Phosphatidylinositol specific phospholipase C treatment of the membrane fraction of the supraoptic homogenate solubilized OBCAM. Kilon, another IgLON member, was also shown to localize at the neurosecretory granules of vasopressin-positive dendrites via the glycosylphosphatidylinositol anchor. High K(+)-stimulation appeared to cause the diffusion of OBCAM-labeled gold particles from neurosecretory granules together with the exocytosis. These findings indicate that OBCAM is synthesized within the somata, attached to vasopressin neurosecretory granules via the glycosylphosphatidylinositol anchor, and transported to the dendrites. Moreover, the subcellular localization of OBCAM is changed in an activity-dependent manner.

Animals↗
PubMed2002

Expression of high levels of tubulin and microtubule-associated protein 2d in the neurohypophysial astrocytes of adult rat.

The hypothalamo-neurohypophysial system, containing arginine vasopressin and oxytocin, is well known to show reversible morphological reorganization for both neurons and glial cells during chronic physiological stimulation. To determine the molecular background for these morphological changes, we investigated the expression of tubulin and microtubule-associated protein (MAP) 2d in the neurohypophysial astrocytes, pituicytes of adult rats by using reverse transcription-polymerase chain reaction, western blot, and immunohistochemistry. The mRNA of MAP2d was expressed at higher levels than that of MAP2c in the neurohypophysis, cerebral cortex, and cerebellum. In contrast, predominant expression of mRNA of MAP2c was detected in the olfactory bulb. Western blot analysis showed the presence of MAP2d in the neurohypophysis, however the amount was below the detection level in the cerebral cortex and cerebellum. A double labeling study using a confocal laser scanning microscope showed intense tubulin immunoreactivity in the glial fibrillary acidic protein (GFAP)-positive pituicytes of the intact neurohypophysis. Almost no tubulin immunoreactivity was observed in the astrocytes of the intact cerebral cortex, cerebellum, and supraoptic nucleus, in contrast to strong tubulin immunoreactivity in neuronal dendrites and somata. Interestingly, intense tubulin immunoreactivity was also observed in the GFAP-positive reactive astrocytes in the immediate vicinity of the artificial lesion of the cerebral cortex. Electron microscopic observation further demonstrated the presence of a lot of microtubules in the pituicytes of intact rats.The present results demonstrate that pituicytes in the adult rat neurohypophysis expresses high levels of tubulin and MAP2d compared with normal brain astrocytes, and suggest that the ability of astrocytic morphological alteration may be at least partly ascribed to this high expression of microtubule proteins.

Animals↗
PubMed2001

Involvement of Ca2+ waves in excitation-contraction coupling of rat atrial cardiomyocytes.

Two-dimensional and line-scan analyses of the early phase Ca2+ transients in rat cardiomyocytes were performed with a rapid-scanning laser confocal microscope and fluo-3 to elucidate the mechanism of activation of Ca2+ release from the sarcoplasmic reticulum in atrial myocytes which lack a well developed T-tubular network. On electrical stimulation of ventricular myocytes, Ca2+ concentration began to rise earliest at the Z-line level and became uniform throughout the cytoplasm within about 10 msec. In contrast, on stimulation of atrial myocytes, the earliest rise in Ca2+ occurred at the cell periphery and then spread to the cell interior; cytoplasmic Ca2+ became uniform after more than 30msec. The velocity of the propagation of rise in Ca2+ was 112 +/- 5.1 microm/sec (n = 10), which was similar to that of spontaneous Ca2+ waves observed in atrial and ventricular myocytes. No difference in frequency, amplitude and kinetics of spontaneous Ca2+ sparks was observed between the subsarcolemmal and central regions of atrial myocytes. Ryanodine concentration-dependently decreased the contractile force of isolated rat atrial and ventricular tissue preparations; the sensitivity was higher in atrial myocytes. The present study visualized the involvement of a propagated Ca2+-induced-Ca+ release mechanism in atrial but not ventricular myocytes. This difference may underlie some of the atrioventricular difference in response to physiological and pharmacological stimuli.

Animals↗
PubMed2001

Phased A-tracts bind to the alpha subunit of RNA polymerase with increased affinity at low temperature.

Previously we showed that the expression of a Clostridium perfringens phospholipase C gene (plc) is activated by promoter upstream phased A-tracts in a low temperature-dependent manner. In this paper we characterize the interaction between the alpha subunit of C. perfringens RNA polymerase and the phased A-tracts. Hydroxyl radical footprinting and fluorescence polarization assaying revealed that the alpha subunit binds to the minor grooves of the phased A-tracts through its C-terminal domain with increased affinity at low temperature. The result provides a molecular mechanism underlying the activation of the plc promoter by the phased A-tracts.

Base Sequence↗
PubMed2001

Hypothalamic 11,12-epoxyeicosatrienoic acid attenuates fever induced by central interleukin-1beta in the rat.

Inhibitors of cytochrome P-450 augment fever in rats and mice, indicating that the metabolite of the enzyme is candidate of endogenous antipyretic. Cytochrome P-450 of arachidonic acid cascade leads to the formation of regioisomeric 5,6-, 8,9- 11,12- and 14,15-epoxyeicosatrienoic acid (EET). Various isomers of EET were administrated into the preoptic area and anterior hypothalamus (PO/AH) to test their influence on fever induced by interleukin-1beta (IL-1beta) administrated into the PO/AH in conscious rats. The IL-beta-induced fever was attenuated in the 11,12-EET-pretreated rats, although 5,6-, 8,9- and 14,15-EET did not affect the fever. Intra-PO/AH injection of 11,12-EET did not alter normal body temperature. The results suggest that 11,12-EET acts in the hypothalamus as an endogenous antipyretic.

8,11,14-Eicosatrienoic Acid↗