Biomedical subjects

Hiroshi Ueda

Publications and source records attributed to Hiroshi Ueda.

At least 91 records · Page 5Linked to original sources

PubMed2003

Year-to-year differences in plasma levels of steroid hormones in pre-spawning chum salmon.

Changes in plasma levels of steroid hormones in pre-spawning chum salmon (Oncorhynchus keta) were examined for 6 years in association with sexual maturation. Fish were sampled along their homing pathway from the coastal sea to the spawning ground from 1995 to 2000. Plasma levels of testosterone (T), 11-ketotestosterone (11KT), estradiol-17beta (E2), 17alpha,20beta-dihydroxy-4-pregnen-3-one (DHP), and cortisol were determined by enzyme immunoassays. Sexual maturity was comprehensively estimated by gonadosomatic indices, histology of gonads, nuptial color, spermiation or ovulation ratio. Since the plasma levels of steroid hormones and sexual maturation differed from year to year, they were compared with year-to-year variation of sea surface temperature (SST) of coastal sea to study influence of oceanographic environment on these physiological data. The SST of the migratory route varied among the years, so that we classified the 6 years into cool, intermediate, and warm years. Concerning maturity, the males that returned to the natal hatchery in the warm years were sexually more advanced than those in the cool years. Furthermore, histological data suggested that final oocyte maturation occurred before arrival at the hatchery in one of the warm years, i.e., 1999, while it occurred at the hatchery in one of the intermediate years, i.e., 2000. In the males, T and 11KT levels increased significantly on midway of the homing route in the warm years, whereas they did not show any noticeable changes in the cool years. Furthermore, the levels of T and 11KT on midway of the homing route in the warm years, i.e., 1998 and 1999, were significantly higher than those in one of the cool years, i.e., 1995, in both sexes. In the females, the levels of E2 decreased during upstream migration. Conversely, those of DHP considerably elevated at spawning ground in all years examined. The levels of cortisol were different from year to year regardless of the SST. The present results showed that there were year-to-year differences in plasma levels of steroid hormones and maturity, and some of them may be influenced by the year-to-year variation of SST.

Animals↗

PubMed2003

An optimized homogeneous noncompetitive immunoassay based on the antigen-driven enzymatic complementation.

We describe an optimized noncompetitive and homogeneous immunoassay based on the antigen-dependent reassociation of antibody variable domains and beta-galactosidase (beta-gal) complementation (open sandwich enzymatic complementation immunoassay, OS-ECIA). The reassociation of two fusion proteins, an antibody heavy chain variable region fragment tethered to an N-terminal deletion mutant of beta-gal, V(H)Deltaalpha, and the light chain variable region fragment tethered to a C-terminal deletion mutant of beta-gal, V(L)Deltaomega, was monitored by the enzymatic complementation between the two. With the use of anti-hen egg lysozyme (HEL) antibody HyHEL10, an antigen-dependent enhancement in the enzymatic activity was clearly observed. To optimize the assay, the lengths of the linkers connecting the two domains of each fusion protein were varied, and the optimal pair V(H)(G(4)S)(2)Deltaalpha/V(L)(G(4)S)Deltaomega showed much improved antigen-responsive beta-gal activity. After various optimizations, almost 1000-fold improvement in sensitivity compared with that of our corresponding homogeneous open sandwich (OS) assays based on the energy transfer was observed, possibly due to lower V(H)/V(L) concentration and background heterodimer association.

Haptens↗

PubMed2003

Neurosteroid-induced hyperalgesia through a histamine release is inhibited by progesterone and p,p'-DDE, an endocrine disrupting chemical.

The intraplantar injection of dehydroepiandrosterone sulfate (DHEAS), a representative neurosteroid, showed hyperalgesia in the Hargreaves' thermal or automatic paw-pressure mechanical nociception test. The DHEAS-induced hyperalgesia was abolished by diphenhydramine (DPH), a H(1) histamine (His) receptor antagonist, as well as the hyperalgesia induced by His or compound 48/80, a mast cell degranulating agent. The DHEAS-induced hyperalgesia was also blocked by progesterone (PROG), another type of neurosteroid and a putative neurosteroid receptor antagonist. Neither DPH nor PROG showed any changes in the thermal threshold. On the other hand, endocrine disrupting chemicals (EDCs) are known to disrupt reproductive system in wild-lives and humans through the disturbance of the endocrine homeostasis. In this study, the flexor responses induced by intraplantar injection of DHEAS were blocked by p,p'-DDE, an EDC as well as by PROG in the algogenics-induced nociceptive flexor responses test (ANF test) in mice. Similarly, p,p'-DDE blocked the DHEAS-induced hyperalgesia in Hargreaves' thermal nociception test. Besides the hyperalgesic actions, DHEAS increased vascular permeability as measured with Evans blue plasma extravasation. Consistent with behavioral studies, it was blocked by DPH, PROG, and p,p'-DDE. These results suggest that DHEAS has significant hyperalgesic and vasodilatory actions through histamine release, and these actions were reversible by PROG and an EDC.

Animals↗

PubMed2003

Emerging functions for tuberoinfundibular peptide of 39 residues.

Tuberoinfundibular peptide of 39 residues (TIP39), a neuropeptide recently purified from the hypothalamus, appears to be an endogenous ligand for the parathyroid hormone 2 (PTH2) receptor. PTH2 receptors are present in several central nervous system and peripheral areas and are particularly concentrated in the hypothalamus, limbic areas and the outer layers of the spinal cord dorsal horn. TIP39-containing neuronal cell bodies have been identified in the subparafascicular area and the medial paralemniscal nucleus, two brainstem regions that project widely through the entire neuraxis. Treatment of hypothalamic explants with TIP39, and intraventricular injection of the peptide, suggest that it might stimulate hypothalamic-releasing factor secretion. Injection of TIP39, and sequestration of endogenous TIP39 by intrathecal injection of an antibody to TIP39, have provided evidence that it is involved in some aspects of pain sensitivity. Thus, TIP39 might be a new neuromodulator.

Amino Acid Sequence↗

PubMed2003

Novel expression of vanilloid receptor 1 on capsaicin-insensitive fibers accounts for the analgesic effect of capsaicin cream in neuropathic pain.

Here, we investigated the mechanism of the antihyperalgesic effect of capsaicin cream in the nerve injury-induced neuropathic pain model in mice. In naive mice, application of capsaicin cream onto footpad caused no significant changes in the thermal latency in contrast to the severe thermal hyperalgesia induced by a capsaicin ointment. On the other hand, application of the cream 3 h before test concentration dependently reversed both thermal and mechanical hyperalgesia observed after partial sciatic nerve injury in mice. In algogenic-induced nociceptive flexion (ANF) test, application of 0.1% capsaicin cream in naive mice blocked intraplantar (i.pl.) nociceptin- and ATP-induced flexion responses, whereas prostaglandin I(2) (PGI(2)) agonist-induced responses were unaffected. After nerve injury PGI(2) agonist-induced flexion responses were hypersensitized, and capsaicin cream concentration dependently blocked these hyperalgesic responses. Intraplantar injection of capsaicin solution in ANF test also produced potent flexion responses in naive mice that were lost after neonatal capsaicin-treatment. Partial sciatic nerve injury in neonatal capsaicin-treated mice caused reappearance of i.pl. capsaicin-induced flexion responses, suggesting novel expression of capsaicin receptors due to injury. The PGI(2) agonist-induced responses were also hypersensitized in such injured mice. Capsaicin cream completely reversed both i.pl. capsaicin- or i.pl. PGI(2) agonist-induced hyperalgesia in neonatal capsaicin-treated injured mice. Finally, novel expression of VR1 receptors on neonatal capsaicin-insensitive neurons after nerve injury was confirmed by immunohistochemistry. The newly expressed VR1 receptors after nerve injury were mainly confined to A-fibers. Together, our results suggest that novel expression of capsaicin receptors in neuropathic condition contributes to the analgesic effects of the capsaicin cream.

Analgesia↗

PubMed2003

Inhibitory effect of Perilla leaf extract and luteolin on mouse skin tumor promotion.

In the present study, the effects of perilla leaf extract (PLE) and luteolin on 7,12-dimethylbenz[a]anthracene (DMBA)- and 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin papillomas in mice were investigated. Topical application of PLE prior to TPA treatment in DMBA-initiated mouse skin resulted in a significant reduction in tumor incidence and multiplicity. An even more potent preventive effect was observed with topical application of luteolin, which we previously identified as an antiinflammatory constituent. PLE was dissolved in drinking water at a 0.05% dose and mice ingested it ad libitum; no significant difference was observed in tumor incidence or multiplicity but there was a significant reduction in tumor volume between the PLE-treated and untreated groups. These results suggest that PLE has potent antipromotion activity and ingesting it as a daily food may provide a beneficial chemopreventive effect.

Animals↗

PubMed2003

[Pain and QOL--morphine-tolerance and morphine-resistant neuropathic pain].

Morphine is now said to have no problematic side effects such as analgesic tolerance and physical dependence for cancer pain patients in clinic, as far as it is appropriately used. However, sub-sensitivity to morphine might be developed when higher doses of morphine are used for terminal cancer pain patients. Along with the severity of cancer, the nature of pain becomes changed to neuropathic pain, which is resistant to morphine or NSAIDS. In order to safely use morphine in the clinic, we need to know how morphine tolerance and neuropathic pain are developed and what adjuvants could be used to completely suppress the pain. Here I overview the proposed mechanisms for morphine tolerance and neuropathic pain in relation to the availability of analgesic adjuvants.

Adrenergic alpha-Antagonists↗

PubMed2003

Molecular mechanism of neuropathic pain.

Pain is the most common clinical symptom for which people seek medical care. Although effective therapy for inflammatory pain is possible, current therapy for neuropathic pain is still inadequate. Even the strongest painkiller, morphine, is reported to be ineffective against neuropathic pain in the clinic. The lack of effective therapeutic agents for neuropathic pain is due to the fact that its underlying mechanisms are still not fully understood. Recently, pain researchers have been trying to identify the cellular and molecular processes that lead to neuropathic pain. Understanding the molecular mechanisms of neuropathic pain would certainly provide a hope for the future development of effective therapies for this intractable painful disease.

Analgesics↗

PubMed2003

Creation of a thermostable firefly luciferase with pH-insensitive luminescent color.

The thermal instability and pH-sensitive spectral property of firefly luciferase have hampered its use as a sensitive multicolor luminescent label or bioluminescent resonance energy transfer donor. With the intention of improving the thermostability of a previously found firefly Hotaria parvula luciferase mutant with minor pH-sensitive spectral change (V368A), further mutation (E356R) was introduced by taking a reportedly stabilized mutant of Photinus pyralis luciferase into account. The double mutant E356R/V368A showed significantly improved thermostability because > 90% activity remained after incubation for 1 h at 45 degrees C, with its specific activity being maintained. Unlike the wild type or V368A, E356R/V368A showed no change in the emission maximum of 568 nm even at pH 6.3, also implying a mutual relationship between thermostability and the proportion of yellow-green luminescent peak under acidic condition.

Animals↗

PubMed2003

Differential gene expression profiles and identification of the genes relevant to clinicopathologic factors in colorectal cancer selected by cDNA array method in combination with principal component analysis.

The clinical outcome of patients with colorectal cancer frequently varies even if they are at the same clinicopathologic stage. Alternative superior tumor markers of colorectal cancer are needed for prediction of clinical outcome. To clarify the regulatory factors in colorectal cancers, we examined differential expression profiles using cDNA macroarray technique with surgically resected specimens obtained from the patients with colorectal cancer. The gene profiles by an average-linkage hierarchical clustering analysis were found to be almost separable into two groups: tumor group and normal mucosa group. The relationship between several clinicopathologic factors and cancer related genes were investigated by using statistical analyses including principal component analysis (PCA). c-myc-binding protein MM-1, and c-jun proto-oncogene were identified as possible markers of tumor histology and clinical prognosis and early growth response protein 1 (EGR1) was selected to play an important role in progression of clinical stage. We conclude that, with PCA method, we successfully selected the genes relevant to clinicopathologic factors using limited population of clinical samples.

Aged↗

PubMed2002

A homogeneous and noncompetitive immunoassay based on the enhanced fluorescence resonance energy transfer by leucine zipper interaction.

Fluorescence resonance energy transfer (FRET) between two GFP variants is a powerful technique to describe protein-protein interaction in a biological system. However, it has a limitation that the two variants tethered to the respective proteins have to be in sufficient proximity upon binding, which is often difficult to attain by simple N- or C-terminal fusions. Here we describe a novel method to significantly enhance FRET between GFP variant-tagged proteins with the use of leucine zippers. For the homogeneous sandwich immunoassay of a high molecular weight antigen human serum albumin (HSA), two separate single-chain Fvs recognizing distant epitopes of HSA were respectively fused with fluorescence donor ECFP or acceptor EYFP, and FRET between the two was analyzed by fluorescence spectrometry. Because these two proteins did not give any detectable FRET uponantigen addition, we tethered each protein with a leucine zipper motif (c-Jun or FosB) at the C-terminus to help the neighborhood of the GFP variants. Upon antigen addition, the new pairs showed significant antigen-dependent FRET. By exchanging the binding domains, the method will find a range of applications for the assay of other proteins and their interactions in vitro or in vivo.

Energy Transfer↗

PubMed2002

Neuropathy-specific analgesic action of intrathecal nicotinic agonists and its spinal GABA-mediated mechanism.

The effect of the nicotinic acetylcholine receptor (nAChR) agonists at the spinal level has not been well studied in neuropathic states. In the present report, we demonstrate the efficacy of intrathecal nicotinic agonists in partial sciatic nerve injury-induced neuropathy model mice. Intrathecal (i.t.) administration of (-)nicotine and (+)epibatidine, at doses without undesirable effects, had no antinociceptive action in sham-operated mice. However, they completely reversed the thermal and mechanical hyperalgesia in partial sciatic nerve-injured mice. This neuropathy-specific analgesic action of i.t. (-)nicotine and (+)epibatidine is attributed to different subtypes of nAChRs at the spinal level. After antagonism experiments with mecamylamine, dihydro-beta-erythroidine and methyllycaconitine, it was observed that (-)nicotine-induced analgesia was mediated through the alpha4beta2 subtype of nAChR while the (+)epibatidine-induced one was mediated through non-alpha4beta2 subtype of nAChR. Moreover, i.t. pretreatment with NMDA receptor antagonist did not block nicotinic analgesia. Similar to nicotinic agonists, gamma-aminobutyric acid receptor (GABA(A)) agonist muscimol (i.t.) produced neuropathy-specific analgesic action giving analgesia only in nerve-injured mice. The GABA(A) antagonists bicuculline and picrotoxin significantly blocked the analgesic effects of muscimol as well as that of (-)nicotine and (+)epibatidine. On the other hand, i.t. injection of nicotinic antagonist mecamylamine and GABA(A) antagonist picrotoxin in sham-operated mice induced a thermal hyperalgesia without any effects in nerve-injured animals suggesting the presence of a tonic inhibitory tone on nociceptive transmission through the spinal cholinergic-GABAergic system. These results also suggest that the neuropathy-specific analgesic action of intrathecal nicotinic agonists was due to stimulation of this cholinergic-GABAergic system whose inhibitory tone had been reduced due to injury.

Analgesics↗

PubMed2002

Molecular cloning and immunohistochemical localization of ubiquitin C-Terminal hydrolase expressed in testis of a teleost, the Nile Tilapia, Oreochromis niloticus.

We previously produced four monoclonal antibodies to testicular proteins of a teleost, the Nile tilapia. One of the monoclonal antibodies, TAT(Testicular Antigen of Tilapia)-10, recognizes a Mr=27,000 protein (27 kD protein), which is present in A and early B type spermatogonia, spermatids, and spermatozoa in testis. In order to clarify the function of this protein, molecular cloning was conducted. The cDNA for the 27 kD protein contains a complete open reading frame encoding 220 amino acid residues. The predicted amino acid sequence of the 27 kD protein was homologous to those of the ubiquitin carboxy-terminal hydrolases (UCH) reported in mammals. The measurement of the ubiquitin-releasing activity of the recombinant 27 kD protein revealed that the protein is the active form of UCH. Northern blot analysis showed that the UCH mRNA was expressed in ovary and brain in addition to the testis. Immunohistochemical study showed that, in brain, UCH was localized especially on the olfactory organ including the olfactory bulb and olfactory epithelium in olfactory rosetta, suggesting the involvement of the protein in chemoreceptive function. In the Tilapia ovary, UCH localized especially in pre-vitellogenic oocytes, suggesting that the enzyme activity could be important in oocyte growth. This is the first report for the cDNA cloning and cellular localization of UCH in fish. J. Exp. Zool. 293:368-383, 2002.

Amino Acid Sequence↗

PubMed2002

Possible mechanisms by which adipocyte lipolysis is enhanced in exercise-trained rats.

A possible mechanism(s) behind exercise training-enhanced lipolysis was investigated in rat adipocytes. Exercise training (9 weeks; running) enhanced the activity of cAMP-dependent protein kinase (PKA) and the protein expressions of PKA subunits (catalytic, RII alpha, and RII beta) in P(40) fraction (sedimenting at 40,000g), but not in I(40) fraction (infranatant of 40,000g) of adipocyte homogenate. The expression of PKA-anchoring protein 150 (AKAP150) in P(40) fraction was greater in exercise-trained (TR) than in control (C) rats. Hormone-sensitive lipase (HSL) activities in both fractions were also greater in TR. On the other hand, stimulated lipolysis was accompanied by increased activities of HSL in P(40) but not in I(40) fraction. The decreases in stimulated lipolysis due to St-Ht31 were greater in TR rats. Thus, the mechanisms behind exercise training-enhanced adipocyte lipolysis could involve the increased activities of PKA and HSL with enhanced expressions of AKAP150 and some subunits of PKA, all of which may be compartmentalized within adipocytes.

Adipocytes↗

PubMed2002

Exercise training increases membrane bound form of tumor necrosis factor-alpha receptors with decreases in the secretion of soluble forms of receptors in rat adipocytes.

We examined the effects of exercise training (treadmill running over 9 weeks) on the ability of isolated adipocytes to secrete tumor necrosis factor-alpha (TNF-alpha) and type 1 soluble TNF receptor (sTNFR1) in vitro in Wistar rats. We also examined the effects of exercise training on the expression of membrane bound forms of type 1 TNF receptor (mTNFR1) in adipocyte crude membranes of the same rat subjects. Exercise training significantly increased the secretions of TNF-alpha from isolated adipocytes. Treatment with a cyclooxygenase inhibitor, either indomethacin (100 microM) or eicosatetraynoic acid (100 microM), significantly blocked the release of TNF-alpha from adipocytes in both exercise-trained rat group and sedentary control rat group, suggesting that some cyclooxygenase metabolite(s) acts as a ligand in TNF-alpha synthesis. Decreased amounts of TNF-alpha were found to be significantly greater in both exercise-trained rat group than in sedentary control rat group after incubation with inhibitors. Thus, the inhibitory effect of both indomethacin and eicosatetraynoic acid was significantly greater in adipocytes from exercise-trained rats. Both plasma sTNFR1 levels and adipocytes-derived sTNFR1 were found to be significantly less in the exercise-trained rat group. Western blot analysis revealed that exercise training remarkably increased the expressions of mTNFR1 in adipocyte crude membrane. Thus, exercise training enhanced the ability of isolated adipocytes to secrete TNF-alpha with reduced secretion of sTNFR1, and provoked the greater expressions of mTNFR1 in adipocyte crude membrane. These alterations may induce enhanced the autocrine effects of TNF-alpha within adipocytes in exercise-trained rats.

5,8,11,14-Eicosatetraynoic Acid↗

PubMed2002

Riluzole enhances expression of brain-derived neurotrophic factor with consequent proliferation of granule precursor cells in the rat hippocampus.

The dentate gyrus of the hippocampus, generating new cells throughout life, is essential for normal recognition memory performance. Reduction of brain-derived neurotrophic factor (BDNF) in this structure impairs its functions. To elucidate the association between BDNF levels and hippocampal neurogenesis, we first conducted a search for compounds that stimulate endogenous BDNF production in hippocampal granule neurons. Among ion channel modulators tested, riluzole, a neuroprotective agent with anticonvulsant properties that is approved for treatment of amyotrophic lateral sclerosis, was highly effective as a single dose by an intraperitoneal injection, causing a rise in BDNF localized in dentate granule neurons, the hilus, and the stratum radiatum of the CA3 region. Repeated, but not single, injections resulted in prolonged elevation of hippocampal BDNF and were associated with increased numbers of newly generated cells in the granule cell layer. This appeared due to promoted proliferation rather than survival of precursor cells, many of which differentiated into neurons. Intraventricular administration of BDNF-specific antibodies blocked such riluzole effects, suggesting that BDNF increase is necessary for the promotion of precursor proliferation. Our results suggest the basis for a new strategy for treatment of memory dysfunction.

Animals↗

PubMed2002

A homogeneous noncompetitive immunoassay for the detection of small haptens.

We describe a noncompetitive homogeneous immunoassay for small haptens based on the antigen-dependent reassociation of antibody variable domains and beta-galactosidase (beta-gal) complementation (open sandwich enzymatic complementation immunoassay). As a model system, the reassociation of two fusion proteins, an anti 4-hydroxy-3-nitrophenylacetyl (NP) antibody heavy-chain variable-region fragment fused to an N-terminal deletion mutant of beta-gal (V(H)delta alpha) and the light-chain variable-region fragment fused to a C-terminal deletion mutant of beta-gal (V(L)delta omega), was monitored by the enzymatic complementation between the two. Upon simple mixing of the reagents with the sample, an antigen (NP)-dependent increase in enzymatic activity was observed. When 5-iodo-NP was measured, a 10 times higher sensitivity was observed, probably due to its higher affinity. Compared with our corresponding heterogeneous open sandwich enzyme-linked immunosorbent assay, approximately 1000-fold improvement in the sensitivity was attained, probably due to lower background V(H)-V(L) association. In addition, the assay required less time, handling, sample volume, and assay reagents.

Electrophoresis, Polyacrylamide Gel↗

PubMed2002

Anatomical and physiological evidence for involvement of tuberoinfundibular peptide of 39 residues in nociception.

The parathyroid hormone 2 (PTH2) receptor's anatomical distribution suggests that, among other functions, it may be involved in modulation of nociception. We localized PTH2 receptor protein to spinal cord lamina II and showed that it is synthesized by subpopulations of primary sensory neurons and intrinsic spinal cord dorsal horn neurons. Tuberoinfundibular peptide of 39 residues (TIP39) selectively activates the PTH2 receptor. Intraplantar microinjection of TIP39 caused a paw-withdrawal response and intrathecal injection caused scratching, biting, and licking, a nocifensive response. Intrathecal administration of a TIP39 antibody decreased sensitivity in tail-flick and paw-pressure assays. Intrathecal administration of TIP39 potentiated responses in these assays. We determined the sequence of TIP39's precursor and found that mRNA encoding TIP39 and TIP39-like immunoreactivity is concentrated in two brainstem areas, the subparafascicular area and the caudal paralemniscal nucleus. Cells in these areas project to the superficial dorsal horn of the spinal cord. Our data suggest that TIP39 released from supraspinal fibers potentiates aspects of nociception within the spinal cord.

Animals↗