Biomedical subjects

M Nishiyama

Publications and source records attributed to M Nishiyama.

At least 19 recordsLinked to original sources

PubMed2006

Purinergic receptor ligands stimulate pro-opiomelanocortin gene expression in AtT-20 pituitary corticotroph cells.

Although recent studies have suggested that purinergic receptors are expressed in the anterior pituitary gland, their involvement in the regulation of pituitary hormone gene expression is not completely understood. In the present study, we examined the expression of purinergic receptors and the effects of purinergic receptor ligands on pro-opiomelanocortin (POMC) gene expression, in AtT20 mouse corticotroph cells. We identified the expression of most of the purinergic receptor subtypes (A1, A2, P2X1, 3-7, P2Y1, 2, 4) mRNAs, analysed by the reverse transcriptase-polymerase chain reaction. We also found that adenosine and ATP, two representative and endogenous agonists of A1-3 and P2X/P2Y receptors, respectively, stimulated the 5'-promoter activity of the POMC gene in a dose- and time-related manner. When these ligands were simultaneously used with corticotrophin-releasing hormone (CRH), effects that were more than additive were observed, suggesting an enhancing role of these compounds in CRH-mediated adrenocorticotrophic hormone (ACTH) synthesis. These ligands also stimulated the expression of transcription factors involved in the regulation of the POMC gene, but did not enhance ACTH secretion. Finally, the positive effect of adenosine as well as CRH was completely inhibited by the protein kinase A inhibitor H89, whereas that of ATP was not influenced, indicating that different intracellular signalling pathways mediate these effects. Altogether, our results suggest a stimulatory role for these purinergic receptor ligands in the regulation of POMC gene expression in corticotroph cells. Because adenosine and ATP are known to be produced within the pituitary gland, it is possible they may be acting in an autocrine/paracrine fashion.

Adenosine↗

PubMed2006

Suppressive effects of dehydroepiandrosterone and the nuclear factor-kappaB inhibitor parthenolide on corticotroph tumor cell growth and function in vitro and in vivo.

Dehydroepiandrosterone (DHEA) is believed to have an anti-tumor effect, as well as anti-inflammatory, antioxidant, and anti-aging effects. To clarify the possible inhibitory action of DHEA on pituitary tumor cells, we tested the effects of DHEA, alone or in combination with the nuclear factor-kappaB (NF-kappaB) inhibitor parthenolide (PRT), on AtT20 corticotroph cell growth and function both in vitro and in vivo. We found that, in vitro, DHEA and PRT had potent inhibitory effects on pro-opiomelanocortin and NF-kappaB-dependent gene expression. They also suppressed the transcription activity of survivin, a representative anti-apoptotic factor, and induced apoptosis in this cell line. Furthermore, using BALB/C nude mice with xenografts of AtT20 cells in vivo, we found that the combined administration of DHEA and PRT significantly attenuated tumor growth and survivin expression. The treatment also decreased the elevated plasma corticosterone levels and ameliorated the malnutrition induced by tumor growth. Altogether, these results suggested that combined treatments of DHEA and PRT potently inhibit the growth and function of corticotroph tumor cells both in vitro and in vivo. This effect may, at least partly, be caused by the suppressive effects of these compounds, such as survivin and other inhibitor of apoptosis proteins, on NF-kappaB-mediated gene transcription.

Adjuvants, Immunologic↗

PubMed2005

Evaluation of different bonded investments for dental titanium casting.

The properties of several different investments were investigated including phosphate bonded, magnesia bonded, and alumina cement investments. Measurements included the setting expansion, thermal expansion, and compressive strength of investments, as well as the tensile strength, elongation, Vickers hardness (VHN) and surface roughness of titanium castings. For phosphate bonded investment, the setting expansion after being mixed with its own mixing solution was 2.10%, which was larger than the other investments; the thermal expansion was -0.25% at 200 degrees C, the compressive strength 14 and 5 MPa after heating. For titanium cast in phosphate bonded investment, the hardness on its top surface was 655 Hv, the tensile strength was 379 MPa, the elongation was 19.4%, and the surface roughness was 2.29 microm. Although the thermal expansion of phosphate bonded investment is small, the setting expansion is large enough to compensate for the shrinkage of titanium castings. As its thermal expansion at T > or = 600 degrees C was constant and its heating-cooling cycle was almost reversible, these two properties can reduce the thermal shock and thus avoid cracking of the investment.

Aluminum Oxide↗

PubMed2004

Management of tight filum terminale.

OBJECTIVE: Tight filum terminale (TFT) is not an uncommon condition, but due to insufficient diagnostic options this syndrome is often overlooked and left untreated. The present study was designed to investigate surgical results concerning neurological perspectives and diagnostic methods for TFT. METHODS: Subjects for this study consisted of 37 patients with TFT who were surgically treated by transecting the filum terminale. We examined the surgical outcomes with regard to low back pain, leg pain, and bladder and bowel dysfunctions, and analyzed neurological and imaging findings. RESULTS: TFT diagnosis was based on the following five criteria; 1) low back pain, 2) non-dermatomal leg pain, 3) bladder-bowel dysfunction, 4) spinal stiffness, and 5) a newly developed positive provocation test for spinal cord extension. CONCLUSION: The survey of surgical outcomes indicated that satisfactory results could be obtained by surgical release of the filum terminale, while neural function was still reversible. In particular, our newly devised provocation test was shown to be highly effective for the early diagnosis of this disorder.

Adolescent↗

PubMed2004

[Mucosa-associated lymphoid tissue lymphoma; report of two cases].

Primary pulmonary malignant lymphoma is a rare disease that is thought to belong to a category of malignant lymphomas arising from mucosa-associated lymphoid tissue (MALT). We presented 2 cases of MALT lymphoma. Case 1: In a 39-year-old woman, an abnormal shadow was detected in chest radiography in the left S3. The diagnosis was organizing pneumonia in consequence of the further examination. One year after an growing abnormal shadow was detected again, we performed the tumor resection with the thoracoscopy in the possibility of malignancy. The pathological diagnosis was MALT lymphoma. Case 2: In a 70-year-old man, middle lobe syndrome was detected in chest radiography. As a result of the bronchial biopsy, the lesion was suspected of MALT lymphoma. Right middle lobectomy was performed, and the tumor was diagnosed as a MALT lymphoma histologically. He underwent an effective postoperative radiotherapy and has had no evidence of recurrence.

Adult↗

PubMed2002

New type of vortex pinning structure effective at very high magnetic fields.

We report on a new type of correlated nanometer-scale pinning structure observed in a melt-processed (Nd0.33Eu0.38Gd0.28)Ba(2)Cu(3)O(y) (NEG-123). It consists of NEG/Ba-rich clusters in the stoichiometric NEG-123 matrix forming a lamellar array with a period of a few nanometers. These lamellas appear within regular twins, thus representing their fine substructure-sometimes straight, sometimes wavy. This new material structure correlates well with the significant enhancement of pinning at high fields, represented by irreversibility field above 14 T at 77 K (B parallel c). We believe that the new pinning medium enables one to significantly broaden the limits for high-field applications.

Journal Article↗

PubMed2001

Cloning of full-length genomic DNA encoding human FcepsilonRI alpha-chain and its transcriptional regulation.

Two novel exons, named exon 1A and exon 2A, were found at 18.4 and 12.6 kb upstream from the exon known as the first exon of human FcepsilonRI alpha-chain gene. Transcription from the promoter present in the upstream of exon 1A was decreased by mutations introduced into the "first intron" between exon 1A and exon 2A, suggesting the presence of an intronic regulatory element in the intron. Consistent with this, electrophoretic mobility shift assay revealed the presence of a nuclear factor which bound the region in FcepsilonRI alpha-chain positive cells.

Base Sequence↗

PubMed2001

Chiral helicity induced by hydrogen bonding and chirality of podand histidyl moieties.

[structure: see text] The single-crystal X-ray structure determination of N,N'-bis[(S)-(+)-1-methoxycarbonyl-2-(4-imidazolyl)ethyl]-2,6-pyridinedicarboxamide (L-BHisPA) and the D-isomer (D-BHisPA) derived from the corresponding chiral histidine revealed a left- and right-handed helical conformation, respectively, through intramolecular hydrogen bonding and chirality of the podand histidyl moieties. Furthermore, each helical molecule is connected by continuous intermolecular hydrogen bonds to afford a left- or right-handed helical assembly, respectively, in the crystal packing.

Journal Article↗

PubMed2001

Characterization of RGS5 in regulation of G protein-coupled receptor signaling.

RGS proteins (regulators of G protein signaling) serve as GTPase-activating proteins (GAPs) for G alpha subunits and negatively regulate G protein-coupled receptor signaling. In this study, we characterized biochemical properties of RGS5 and its N terminal (1-33)-deleted mutant (deltaN-RGS5). RGS5 bound to G alpha(i1), G alpha(i2), G alpha(i3), G alpha(o) and G alpha(q) but not to G alpha(s) and G alpha13 in the presence of GDP/AIF4-, and accelerated the catalytic rate of GTP hydrolysis of G alpha(i3) subunit. When expressed in 293T cells stably expressing angiotensin (Ang) AT1a receptors (AT1a-293T cells), RGS5 suppressed Ang II- and endothelin (ET)-1-induced intracellular Ca2+ transients. The effect of RGS5 was concentration-dependent, and the slope of the concentration-response relationship showed that a 10-fold increase in amounts of RGS5 induced about 20-25% reduction of the Ca2+ signaling. Furthermore, a comparison study of three sets of 293T cells with different expression levels of AT1a receptors showed that RGS5 inhibited Ang II-induced responses more effectively in 293T cells with the lower density of AT1a receptors, suggesting that the degree of inhibition by RGS proteins reflects the ratio of amounts of RGS proteins to those of activated G alpha subunits after receptor stimulation by agonists. When expressed in AT1a-293T cells, deltaN-RGS5 was localized almost exclusively in the cytosolic fraction, and exerted the inhibitory effects as potently as RGS5 which was present in both membrane and cytosolic fractions. Studies on relationship between subcellular localization and inhibitory effects of RGS5 and deltaN-RGS5 revealed that the N terminal (1-33) of RGS5 plays a role in targeting this protein to membranes, and that the N terminal region of RGS5 is not essential for exerting activities.

Angiotensin II↗

PubMed2001

Histone deacetylase as a new target for cancer chemotherapy.

Trichostatin A (TSA) and trapoxin (TPX), inhibitors of the eukaryotic cell cycle and inducers of morphological reversion of transformed cells, inhibit histone deacetylase (HDAC) at nanomolar concentrations. Recently, FK228 (also known as FR901228 and depsipeptide) and MS-275. antitumor agents structurally unrelated to TSA, have been shown to be potent HDAC inhibitors. These inhibitors activate the expression of p21Waf1 in a p53-independent manner. Changes in the expression of regulators of the cell cycle, differentiation, and apoptosis with increased histone acetylation may be responsible for the cell cycle arrest and antitumor activity of HDAC inhibitors. TSA has been suggested to block the catalytic reaction by chelating a zinc ion in the active site pocket through its hydroxamic acid group. On the other hand, an epoxyketone has been suggested to be the functional group of TPX capable of alkylating the enzyme. We synthesized a novel TPX analogue containing a hydroxamic acid instead of the epoxyketone. The hybrid compound, called cyclic hydroxamic-acid-containing peptide 1 (CHAP1) inhibited HDAC at low nanomolar concentrations. The HDAC1 inhibition by CHAPI was reversible, as is that by TSA, in contrast to irreversible inhibition by TPX. Interestingly, HDAC6, but not HDAC1 or HDAC4, was resistant to TPX and CHAP1, while TSA inhibited these HDACs to a similar degree. CHAP31, the strongest HDAC inhibitor obtained from a variety of CHAP derivatives, exhibited antitumor activity in BDF1 mice bearing B16/BL6 tumor cells. These results suggest that CHAP31 is promising as a novel therapeutic agent for cancer treatment, and that CHAP may serve as a basis for new HDAC inhibitors and be useful for combinatorial synthesis and high-throughput screening.

Animals↗

PubMed2001

Phototrophic N2 fixation suppressed by activated sulfate reduction in anoxic rice soil slurries.

Interactions between sulfate reduction (SR) and phototrophic nitrogenase activities were investigated in rice soil slurries mixed with rice straw. Activation of SR by adding exogenous sulfate suppressed acetylene-reducing activity (ARA) of the slurries, which was associated with phototrophic purple bacteria (PB) enumerated to 108-109 MPN g-1 dry weight (dw) soil. Adding 5 mm sodium molybdate, an inhibitor of SR, markedly increased ARA. However, in the slurries receiving both molybdate and exogenous sulfate, the effects declined simultaneously with partial recovery of SR. These results indicate outcompetition of sulfate-reducing bacteria (SRB) with PB in rice soil, when sulfate concentrations are high enough to support SR. The increasing effects of molybdate on ARA continued during the incubation in the sulfate-depleted condition, probably because of absence of SR and toxicity of molybdate to methanogenesis. Accordingly, stopping activities of the competitive microorganisms may be efficient to increase N2 fixation in rice soil.

Enzyme Inhibitors↗

PubMed2001

Limb-lengthening and angular correction for dysplasia epiphysealis hemimelica.

We report a patient with severe varus deformity and tibial shortening caused by dysplasia epiphysealis hemimelica (DEH), a rare skeletal developmental disorder characterized by an osteocartilaginous tumor arising from an epiphysis in young children. Limb lengthening and angular correction was performed successfully, with an excellent result without abnormal bone growth.

Bone Lengthening↗

PubMed2001

Senescent cells are resistant to death despite low Bcl-2 level.

Cellular senescence may be an antioncogenic event. Bcl-2 is a product of the oncogene, bcl-2, but it may also participate in cellular senescence. To investigate the role of Bcl-2, we analyzed the level of Bcl-2 during aging of normal human fibroblasts by immunoblot analysis and found that its level was highly suppressed in four normal senescent fibroblast strains. This result suggests that senescent cells are more sensitive to induction of death than young cells. Nevertheless, senescent cells showed more resistance to death induced by hydrogen peroxide than young cells, according to LDH assay. Because the balance among antiapoptotic and proapoptotic proteins is an important factor for the determination of cell death, we examined levels of other Bcl-2 family and apoptosis-regulating proteins, but observed no reasonable change to explain the resistance. During the course of the death induction experiment, we obtained results showing that growth-arrested cells were also resistant to death, and this was further confirmed by a BrdU-labeling experiment. As senescent cells are stopped permanently in the G0/G1-phase of the cell cycle, our data strongly suggest that this is the cause of resistance to death in senescent cells.

Apoptosis↗

PubMed2001

Sodic soils reclaimed with by-product from flue gas desulfurization: corn production and soil quality.

Interest is growing in the use of by-product from flue gas desulfurization (FGD) to reclaim sodic soils by controlling the pH and excessive Na+. This study evaluated the effects on corn (Zea mays) production and pH and electrical conductivity (EC) of calcareous sodic soil during four times of cultivation when the by-product was applied once at the first cultivation (Study I) and the impacts on plant and soil quality at first cultivation when the by-product was applied to the soil at 23,000 kg ha-1 (Study II). In Study I, the germination rate and corn production increased by applying the by-product (0, 5,800, 11,600, and 23,100 kg ha-1), and the greatest total amounts of corn production during the four times of cultivation was when the by-product was applied at 23,100 kg ha-1. In Study II, the pH, exchangeable sodium percentage (ESP), clay dispersion and soluble Na+ in the soil decreased and soluble Mg2+ and soluble K+ in the soil increased. The soil pH was reduced from 9.0 to 7.7 by applying the by-product. However, the by-product decreased the concentrations of total N and P in corn leaves in this study. No significant difference in the concentrations of Mo, Zn, Pb, Ni, Cd, Mn, Cr, Cu, and Al in corn leaves and the soil was observed between the by-product addition and the control except for B in the soil and Fe in corn leaves. The concentration of B in the soil was reduced from 28.7 mg kg-1 to 25.4 mg kg-1 and the concentration of Fe in corn leaves increased from 17.5 mg kg-1 to 22.6 mg kg-1 by applying the by-product in our study.

Conservation of Natural Resources↗

PubMed2001

Substeps within the 8-nm step of the ATPase cycle of single kinesin molecules.

Kinesin is a molecular motor that moves processively by regular 8-nm steps along microtubules. The processivity of this movement is explained by a hand-over-hand model in which the two heads of kinesin work in a coordinated manner. One head remains bound to the microtubule while the other steps from the alphabeta-tubulin dimer behind the attached head to the dimer in front. The overall movement is 8 nm per ATPase cycle. To investigate elementary processes within the 8-nm step, we have developed a new assay that resolves nanometre displacements of single kinesin molecules with microsecond accuracy. Our data show that the 8-nm step can be resolved into fast and slow substeps, each corresponding to a displacement of approximately 4 nm. The substeps are most probably generated by structural changes in one head of kinesin, leading to rectified forward thermal motions of the partner head. It is also possible that the kinesin steps along the 4-nm repeat of tubulin monomers.

Animals↗

PubMed2001

Differential neuropeptide responses to starvation with ageing.

During starvation, counterregulatory responses to loss of food (i.e. responses that lead to an increase in appetite) occur in the central nervous system (CNS). This study was designed to examine whether middle-aged rats show greater or smaller behavioural, peripheral and central hormonal responses during starvation compared to young rats. In experiment 1, refeeding following 4 days of starvation was measured in both middle-aged (72-week-old) and young (9-week-old) rats. The level of refeeding was similar to each prestarved level until 3 days after the end of starvation in both groups. From the 4th day, the level of refeeding in young rats increased and reached beyond the prestarved level, whereas refeeding in middle-aged rats remained similar to the prestarved level. Thus, overall refeeding throughout 7 days was greater in young rats than in middle-aged rats. In experiment 2, middle-aged and young rats were starved for 4 days and were killed in the morning. Middle-aged rats showed a smaller plasma corticosterone response than that of young rats. The magnitude of decreases in plasma glucose, insulin and leptin was similar in both groups. In the arcuate nucleus, the starvation-induced increase in neuropeptide Y (NPY) mRNA and the decrease in proopiomelanocortin (POMC) mRNA were smaller in middle-aged rats than in young rats. In contrast, the starvation-induced decrease in corticotrophin-releasing hormone (CRH) mRNA in the hypothalamic paraventricular nucleus was greater in middle-aged rats than young rats. The magnitude of decrease in type-2 CRH receptor mRNA in the ventromedial hypothalamus was similar in both groups. The results indicate that (a) ageing impaired refeeding response (b), middle-aged rats showed the same directional neuropeptide mRNA responses as seen in young rats during starvation and (c) the magnitude of these counterregulatory responses in the CNS in middle-aged versus young rats was not uniform, but rather was site-specific or neuropeptide-specific. This study suggests the importance of NPY and POMC responsiveness in the arcuate nucleus in the age-related differences resulting from starvation-induced refeeding.

Adrenocorticotropic Hormone↗

PubMed2001

Replacements of amino acid residues at subsites and their effects on the catalytic properties of Rhizomucor pusillus pepsin, an aspartic proteinase from Rhizomucor pusillus.

Site-directed mutagenesis was carried out to investigate the functional roles of amino acid residues of Rhizomucor pusillus pepsin (RMPP) in substrate-binding and catalysis. Mutations of two amino acid residues, E13 in the S3 subsite and N219 in the S3/S4 subsites, caused marked changes in kinetic parameters for two substrate peptides with different sequences. Further site-directed mutagenesis at E13 suggested that E13 plays a critical role in forming the correct hydrogen bond network around the active center. In the crystal structure of Rhizomucor miehei pepsin (RMMP), which is an aspartic proteinase produced by Rhizomucor miehei and shows 81% amino acid identity to RMPP, the Oepsilon atom of N219 forms a hydrogen bond with the N-H of isovaline in pepstatin A, a statine-type inhibitor, at the P3 position, suggesting that the loss of the hydrogen bond causes an unfavorable arrangement of the P3 residue. Among the mutants constructed, the E13A mutant showed a 5-fold increase in the ratio of clotting versus proteolytic activity without significant loss of clotting activity. This mutant may present a promising candidate for a useful milk coagulant.

Amino Acid Substitution↗

PubMed2001

Active transepithelial transport of irinotecan (CPT-11) and its metabolites by human intestinal Caco-2 cells.

Irinotecan (CPT-11) is a camptothecin analog with low (about 10--20%) and variable oral bioavailability in animal models. Here, Caco-2 cells were used to evaluate the transepithelial transport of CPT-11 and its metabolites. Caco-2 cells demonstrated significant expression of P-glycoprotein (P-gp), multidrug resistance-associated protein and canalicular multispecific organic anion transporter. Both the lactone and carboxylate forms of CPT-11 and SN-38 were actively transported across the cell monolayers, mainly by the apical-localized P-gp pump. Cellular permeability of CPT-11 at a concentration of 17 microM converted from active to passive-diffusional transport between the 2 and 6 h exposure time points. Antiproliferative effects of CPT-11 were related to permeability of the lactone form, whereas for SN-38 efficacy was dependent on lactone accumulation. Exposure of CPT-11 with cyclosporin A significantly enhanced its efficacy, whereas this was not observed with verapamil and R101933. In contrast, SN-38 efficacy decreased in the presence of P-gp inhibitors due to active transport toward the basolateral side, thereby reducing drug accumulation. Hence, multiple-active transport systems could be demonstrated to be responsible for not only accumulation profiles but also cytotoxic efficacy of CPT-11 and SN-38 in the intestinal Caco-2 cells. It is suggested that CPT-11 might act in a time-dependent manner and that SN-38-mediated cytotoxicity relates to (dose-dependent) lactone kinetics. The results detailed in this report could contribute toward the development of a clinically useful oral formulation of CPT-11 with improved absorption characteristics and suggest that cyclosporin A is a suitable agent for further research of this concept.

ATP Binding Cassette Transporter, Subfamily B, Mem↗