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Biomedical subjects

S Uchida

Publications and source records attributed to S Uchida.

At least 109 records · Page 6Linked to original sources

Calcitonin gene-related peptide produces skeletal muscle vasodilation following antidromic stimulation of unmyelinated afferents in the dorsal root in rats.

In anesthetized rats, the contribution of calcitonin gene-related peptide (CGRP) to antidromic vasodilation of skeletal muscle blood flow (MBF) following electrical stimulation of muscle afferent was investigated by measuring biceps femoris MBF using laser Doppler flowmetry. Repetitive antidromic electrical stimulation of unmyelinated C fibers in ipsilateral dorsal roots at the 3rd-5th lumbar segments for 30 s caused an increase in MBF for 3-15 min (mean 4.5 min) without significant change in systemic arterial blood pressure. The increase in skeletal MBF started about 10 s after the onset of stimulation, and peaked at approximately 130% of the control value at about 30 s after the end of the 30 s period of stimulation. The MBF response was totally abolished by topical application of hCGRP (8-37), a CGRP receptor antagonist. It is concluded that antidromic vasodilation in skeletal muscles following stimulation of unmyelinated C afferents in dorsal roots is independent of systemic blood pressure and is mediated essentially by CGRP. It is suggested that this CGRP-related antidromic vasodilation may be important in the clinical improvement of skeletal MBF produced by physical therapy, e.g. acupuncture.

Afferent Pathways↗

The suppression of small GTPase rho signal transduction pathway inhibits angiogenesis in vitro and in vivo.

Angiogenesis consists of multistep pathways such as the degradation of the matrix, proliferation of the endothelial cells, motility of the endothelial cells, formation of the cord structure and network formation of microvessels. The small GTPase Rho participates in cell motility through actin fiber polymerization. The role of the small GTPase Rho signal transduction pathway in regulating angiogenesis, however, is still unknown. In this study, we investigated the role of the small GTPase Rho signal transduction pathway in angiogenesis in vitro and in vivo using the exoenzyme, Clostridium botulinum C3 transferase, which specifically suppresses Rho and a compound, Y-27632, which suppresses p160ROCK (Rho-associated coiled-coil containing protein kinase). In this paper, we showed that the small GTPase Rho-p160ROCK signal transduction pathway played an important role in angiogenesis both in vitro and in vivo. These results suggest that inhibition of the small GTPase Rho signal transduction pathway by the p160ROCK inhibitor could be a possible new strategy for angiogenic diseases.

Amides↗

Electro-acupuncture stimulation to a hindpaw and a hind leg produces different reflex responses in sympathoadrenal medullary function in anesthetized rats.

The effects of electro-acupuncture stimulation (EAS) of two different areas of a hindlimb with different stimulus intensities on sympathoadrenal medullary functions were examined in anesthetized artificially ventilated rats. Two needles of 160 microm diameter and about 5 mm apart were inserted about 5 mm deep into a hindpaw (Chungyang, S42) or a hind leg (Tsusanli, S36) and current of various intensities passed to excite various afferent nerve fiber groups at a repetition rate of 20 Hz and pulse duration of 0.5 ms for 30-60 s. Fiber groups of afferent nerves stimulated in a hindlimb were monitored by recording evoked action potentials from the afferents innervating the areas stimulated. The sympathoadrenal medullary functions were monitored by recording adrenal sympathetic efferent nerve activity and secretion rates of catecholamines from the adrenal medulla. EAS of a hindpaw at a stimulus strength sufficient to excite the group III and IV somatic afferent fibers produced reflex increases in both adrenal sympathetic efferent nerve activity and the secretion rate of catecholamines. EAS of a hind leg at a stimulus strength sufficient to excite the group III and IV afferent fibers produced reflex responses of either increases or decreases in sympathoadrenal medullary functions. All responses of adrenal sympathetic efferent nerve activity were lost after cutting the afferent nerves ipsilateral to the stimulated areas, indicating that the responses are the reflexes whose afferents nerve pathway is composed of hindlimb somatic nerves. It is concluded that electro-acupuncture stimulation of a hindpaw causes an excitatory reflex, while that of a hind leg causes either excitatory or inhibitory reflex of sympathoadrenal medullary functions, even if both group III and IV somatic afferent fibers are stimulated.

Action Potentials↗

Increase in receptor binding affinity for nimodipine in the rat brain with permanent occlusion of bilateral carotid arteries.

The permanent occlusion of bilateral common carotid arteries (2VO) in rats has been shown to cause progressive and long-lasting cognitive deficits which may be due to impairment of memory retention and/or memory recall process. To clarify the function of voltage dependent calcium channels and the receptor binding of nimodipine by chronic cerebral ischemia, we examined specific (+)-[3H]PN 200-110 binding and the effect of oral administration of nimodipine in brain regions and hearts of rats, at 2 weeks to 4 months after permanent 2VO. There was no significant difference in either dissociation constant (Kd) or maximal number of binding sites (Bmax) for (+)-[3H]PN 200-110 in the cerebral cortex, hippocampus, corpus striatum and thalamus between 2VO and sham rats. In addition, in vitro inhibitory effect of nimodipine on cerebral cortical (+)-[3H]PN 200-110 binding in 2VO rats was similar to that in sham rats. Compared to control rats, oral administration of nimodipine to both 2VO and sham rats at 2 months after permanent 2VO brought about a significant increase in Kd values of specific (+)-[3H]PN 200-110 binding in the cerebral cortex, hippocampus, thalamus and myocardium, and the increase in Kd values was much larger in brain regions of 2VO rats than sham rats. However, the increase in Kd values in the myocardium did not differ between 2VO and sham rats. This observation suggests an increased in vivo binding affinity for nimodipine in chronic ischemic brain. In conclusion, the present study has shown that oral administration of nimodipine may cause a greater occupation in vivo of 1,4-dihydropyridine (DHP) calcium channel antagonist receptors in brains of permanent 2VO rats than in sham rats. Thus, nimodipine may be pharmacologically effective in preventing brain dysfunction due to cerebral ischemia in vivo.

Animals↗

Antibody against a novel, myriocin (ISP-I)-based immunosuppressant, FTY720.

An antibody was prepared by immunizing rabbits with an ovalbumin conjugate of 2-amino-2-(2-(4-(4-mercaptobutyl)phenyl)ethyl)propane-1,3-diol HCl (AMPD-4), which contains the essential structure of the novel immunosuppressant FTY720. As the antibody reacted to not only AMPD-4, but also FTY720, it should be useful for immunoassay of FTY720 in body fluids, tissues and cells.

Animals↗

Imaging the effects of individual zinc impurity atoms on superconductivity in Bi2Sr2CaCu2O8+delta

Although the crystal structures of the copper oxide high-temperature superconductors are complex and diverse, they all contain some crystal planes consisting of only copper and oxygen atoms in a square lattice: superconductivity is believed to originate from strongly interacting electrons in these CuO2 planes. Substituting a single impurity atom for a copper atom strongly perturbs the surrounding electronic environment and can therefore be used to probe high-temperature superconductivity at the atomic scale. This has provided the motivation for several experimental and theoretical studies. Scanning tunnelling microscopy (STM) is an ideal technique for the study of such effects at the atomic scale, as it has been used very successfully to probe individual impurity atoms in several other systems. Here we use STM to investigate the effects of individual zinc impurity atoms in the high-temperature superconductor Bi2Sr2CaCu2O8+delta. We find intense quasiparticle scattering resonances at the Zn sites, coincident with strong suppression of superconductivity within approximately 15 A of the scattering sites. Imaging of the spatial dependence of the quasiparticle density of states in the vicinity of the impurity atoms reveals the long-sought four-fold symmetric quasiparticle 'cloud' aligned with the nodes of the d-wave superconducting gap which is believed to characterize superconductivity in these materials.

Journal Article↗

Suppression of gamma activity in the human medial temporal lobe by sevoflurane anesthesia.

We have reported the presence of continuous gamma (30-150 Hz) activity in the human medial temporal lobe (MTL). Since the MTL is involved in learning and memory, we speculated that MTL gamma activity is related to such higher brain functions. It is thus of interest to learn how this activity changes during different states of consciousness. In this study, we recorded electrocorticographic (ECoG) activity directly from the surface of the MTL after various doses of sevoflurane anesthesia. Five epileptic patients underwent electrode placement operations in which electrodes were attached to the surfaces of the MTL and the basal temporal lobe (BTL). Immediately following the operation ECoG was recorded from each patient under four concentrations of sevoflurane anesthesia (1.5, 2.0, 2.5 and 3.0%). Fast Fourier Transform (FFT) analysis was performed on the MTL ECoGs. Under the lowest sevoflurane concentration, MTL gamma activity was observed in all patients. However, gamma activity was progressively suppressed by increased concentrations of sevoflurane, in a dose-dependent manner. Sevoflurane has been known to reduce neuronal excitability in the rat hippocampus in vitro, probably by changing GABAergic inhibition. The reduction of MTL gamma in the present study may be the result of such a mechanism. Although memory function was not tested in this study, the amount of MTL gamma activity may be related to residual memory function during anesthesia.

Adolescent↗

Dopamine activates inward rectifier K+ channel in acutely dissociated rat substantia nigra neurones.

The effect of dopamine (DA) was investigated on acutely dissociated rat substantia nigra pars compacta (SNc) neurones by using patch clamp recording. The SNc neurones could be classified into two groups. About 75% of large neurones (>30 microm in diameter) were tyrosine hydroxylase (TH) positive while almost all small neurones (<20 microm) were TH negative. In the large neurones, DA hyperpolarized the membrane, resulting in a reduction of the frequency of spontaneous action potentials in current-clamp mode and induced an inward rectifier K+ current in voltage-clamp mode. Quinpirole, a D2 receptor agonist, mimicked the DA action. S(-)-sulpiride, a D2 receptor antagonist, inhibited the DA-induced current (I(DA)) more effectively than SKF83566, a D1 receptor antagonist. Intracellular application of either guanosine 5'-O-(2-thiodiphosphate) (GDP-betaS) or pertussis toxin (IAP) suppressed I(DA). Guanosine 5'-O-(3-thiotriphosphate) (GTP-gammaS) sustained the DA response. Modulators for cAMP such as forskolin and isobutylmethylxathine, H-89, a protein kinase A inhibitor, and chelerythrine, a protein kinase C inhibitor, had no effect on I(DA). The frequency of DA-induced single channel currents in the inside-out patch configuration, for which the unitary conductance was 56.6pS, was greatly reduced by the replacement of GTP with GDP perfused at the cytosolic side. These results suggest that DA acts on a D2-like receptor and activates directly an IAP-sensitive G protein coupled with inward rectifier K+ channels, resulting in a decrease in the spontaneous firing activities of rat SNc dopaminergic neurones.

Animals↗

Separation of Tc-99 in soil and plant samples collected around the chernobyl reactor using a Tc-selective chromatographic resin and determination of the nuclide by ICP-MS

Technetium (Tc) is known to have high mobility in a soil-water system and also high bioavailability for plants, because the most stable form of Tc in natural surface environment is thought to be TcO4- which is highly soluble. The chemical form of Tc, however, changes with environmental conditions. Thus, it is necessary, for realistic assessment, to obtain transfer parameters, such as transfer factors, under natural conditions. However, it is difficult to obtain these parameters using global fallout 99Tc in actual fields due to its low concentration. In this study, 99Tc concentrations in surface soil and plant leaf samples collected from forest sites within the 30-km zone around the Chernobyl reactor were measured for the first time. In the case of soil samples, a simple and rapid analytical method for determination of 99Tc is used which consists of volatilizing and trapping Tc in a combustion apparatus, purifying the Tc with an extraction chromatographic resin, and measuring it by inductively coupled plasma mass spectrometry (ICP-MS). For plant samples, a wet digestion method in combination with the resin is applied and the 99Tc is measured by ICP-MS. Concentrations of 99Tc in organic soil samples and leaves of strawberry (Fragaria vesca) range from 1.1-14.8 Bq kg(-1) dry weight and 0.2-6.0 Bq kg(-1) dry weight, respectively. Our results indicate that soil-to-plant transfer factors for Tc are similar to those for Cs.

Journal Article↗

Measurement of technetium-99 in Marshall Islands soil samples by ICP-MS

Extraction techniques for recovery of technetium-99 (99Tc) for Inductively Coupled Plasma Mass Spectrometry (ICP-MS) measurements were evaluated using soil samples collected from the Marshall Islands. The results of three different extraction techniques were compared: (MI) acid leaching of Tc from ashed soil; (M2) acid leaching of Tc from raw dry soil; and (M3) Tc volatilization from ashed soil using a combustion apparatus. Total Tc recoveries varied considerably between the extraction techniques but each method yielded similar analytical results for 99Tc. Applications of these extraction techniques to a series of environmental samples and ICP-MS measurements have yielded first data on the 99Tc content of Marshall Islands soil samples contaminated with close-in radioactive fallout from nuclear weapons testing. The 99Tc activity concentration in the soil samples ranged between 0.1 and 1.1 mBq g(-1) dry weight (dw). The limit of detection for 99Tc by ICP-MS was 0.17 mBq per sample or 0.014 mBq g(-1) dw under standard operating conditions.

Journal Article↗

Transient expression of activin betaA mRNA on osteoprogenitor cells in rat bone regeneration after drill-hole injury.

We investigated the expression of activin betaA on osteoprogenitor cells in the regenerating bone and bone marrow of the rat femur after drill-hole injury, by immunocytochemistry and in situ hybridization. The periosteum and endosteum adjacent to the wound region showed marked thickening at day 3 and abundant osteoprogenitor cells, which were immunoreactive for proliferating cell nuclear antigen and showed positive reactions for alkaline phosphatase activity, and existed in the inner layer of the periosteum as well as in the endosteum. During the same period, these osteoprogenitor cells began to exhibit activin betaA immunoreactivity and mRNA expression. However, the latter expression gradually reduced the intensity as the cells started to express osteocalcin mRNA during their differentiation to osteoblasts participating in the periosteal and medullary bone formation from day 5. Immunoreactivity for activin type IB and II receptors was also found on activin betaA-immunoreactive cells between days 3 and 7. The above findings suggest that proliferating osteoprogenitor cells, before their transformation to osteoblasts, transiently produce and release activin A, which may play crucial roles in bone and bone marrow regeneration in a receptor-mediated, autocrine and paracrine fashion.

Animals↗

In vivo specific binding characteristics and pharmacokinetics of a 1,4-dihydropyridine calcium channel antagonist in the senescent mouse brain.

PURPOSE: To characterize the in vivo specific binding and pharmacokinetics of a 1,4-dihydropyridine (DHP) calcium channel antagonist, PN 200-110, in the senescent brain, using senescence-accelerated prone mice (SAMP8) and senescence-resistant mice (SAMR1). METHODS: Blood, brain, and heart samples were taken periodically from SAMR1 and SAMP8 following intravenous injection of (+)-[3H]PN 200-110, and the concentration of (+)-[3H]PN 200-110 in the plasma and tissues was determined. In addition, the in vivo specific binding of (+)-[3H]PN 200-110 in the brains of SAMRI and SAMP8 was measured periodically after intravenous injection of the radioligand. RESULTS: There was very little significant difference between SAMR1 and SAMP8 in terms of the half-life (t(1/2)), total body clearance (CL(tot)), steady-state volume of distribution (Vd(ss)). and AUC for the plasma concentration of (+)-[3H]PN 200-110 after intravenous injection of the radioligand. The brain concentration (AUCbrain) for (+)-[3H]PN 200-110 and the brain/plasma AUC ratio (AUCbrain/AUCplasma) were significantly lower in SAMP8 than in SAMR1, and the heart concentration (AUCheart) and the heart/plasma AUC ratio (AUCheart/AUCplasma) were similar in both strains. Also, the brain/plasma unbound AUC ratio (AUCbrain/AUCplasma-free) for (+)-[3H]PN 200-110 was significantly lower in SAMP8 than in SAMRI. The in vivo specific binding (AUCspecific binding, maximal number of binding sites: Bmax) of (+)-[3H]PN 200-110 was significantly lower in brain particulate fractions of SAMP8 than SAMR1. CONCLUSIONS: The concentration and in vivo specific binding of (+)-[3H]PN 200-110 was significantly reduced in the senescent brain. The simultaneous analysis of the concentrations of centrally acting drugs and the in vivo specific binding in the brain in relation to their pharmacokinetics may be valuable in evaluating their CNS effects.

Aging↗

Soluble Flt-1 gene therapy for peritoneal metastases using HVJ-cationic liposomes.

Many studies have reported a close association between VEGF and tumor angiogenesis. The aim of the present study was to evaluate the effectiveness of gene therapy against cancer, including peritoneal metastasis, using a cDNA encoding a soluble type of Flt-1, one of the VEGF receptors. In a peritoneal metastasis model of MKN45 human gastric cancer cells, mice repetitively treated with intraperitoneal injections of HVJ-Fex, a type of HVJ-cationic liposome encapsulating a plasmid expressing soluble mFlt-1, exhibited smaller disseminated foci with fewer microvessels, thus resulting in a significantly longer survival period than the control mice. In another peritoneal metastasis model using HT1080S cells, a clone of HT1080 human fibrosarcoma cells stably transfected with hVEGF, treatments with HVJ-Fex also reduced the growth of disseminated foci without ascites formation. In conclusion, this study demonstrated that the peritoneal metastases of some cancers were largely dependent on VEGF, and that the repeated intraperitoneal transduction of a soluble flt-1 gene using HVJ-cationic liposomes suppressed peritoneal metastases, thereby contributing to a longer survival period.

Animals↗

Mothers' wakefulness at night in the post-partum period is related to their infants' circadian sleep-wake rhythm.

The relationship between a post-partum mother's wakefulness at night and her infant's circadian sleep-wake rhythm was examined. The subjects were seven primipara and their infants. Actigraphic recordings for the mothers and their infants were made over three to five continuous days during Weeks 3, 6, 9 and 12. A 24-h peak of autocorrelograms of the infants' movements appeared in two cases at Week 6, in six cases at Week 9, and in seven cases at Week 12. The mothers' night-time movements significantly decreased from Week 3 to Week 12. Mothers' wakefulness during night sleep in the post-partum period is related to their infants' obtaining circadian sleep-wake rhythm.

Adult↗

GVHD after transfusion of stored RBC concentrates in a solution of mannitol, adenine, phosphate, citrate, glucose, and NaCl following trauma.

BACKGROUND: It has not previously been reported that WBC-reduced RBC preparations can cause transfusion-associated GVHD, even in an immunocompetent individual. CASE REPORT: A 74-year-old man suffered a hemorrhage from the mesentery of the transverse colon after a traffic accident. During surgery, he received 10 units of RBCs from 10 donors in a solution containing mannitol, adenine, phosphate, citrate, glucose and NaCl (MAP). MAP RBCs had been stored for 7 to 8 days before use. On the 27th day after surgery, an erythematous, pruritic rash appeared over the face, neck, and trunk, which was associated with low-grade fever and pancytopenia. Transfusion-associated GVHD was strongly suspected and was confirmed by skin biopsy. To determine the origin of lymphocytes causing GVHD, several microsatellite loci were amplified from DNA of the patient's nails and blood and from blood samples of all 10 RBC donors by using PCR. Amplified alleles derived from the patient's blood were identical to those from one of the 10 samples. CONCLUSION: These findings indicate that transfusions of MAP-RBCs can cause transfusion-associated GVHD in an elderly but immunocompetent host.

Accidents, Traffic↗

Simple laparoscopic ultrasound technique for prevention of bile duct injuries.

BACKGROUND: Bile duct injuries are serious complications of laparoscopic cholecystectomy. Laparoscopic ultrasonography (LUS) has been utilized over the last several years to screen for bile duct calculi and to delineate biliary anatomy. We have found a simple LUS scanning technique that can be useful for preventing bile duct injuries. METHOD: After initial scanning for screening, laparoscopic dissection is continued, isolating the cystic duct. If necessary, scanning can be performed to assure the location of the cystic duct before clipping. After clips are applied to the cystic duct, prior to its incision or transection, LUS is repeated to examine the cystic and bile ducts. RESULTS: This postclipping study can confirm that the clips are applied to the cystic duct and that the hepatic and common bile ducts are intact without occlusion. CONCLUSION: This additional LUS scanning maneuver is simple and quick and may help prevent bile duct injuries.

Bile Ducts↗

Physiological role of CLC-K1 chloride channel in the kidney.

Although CLC chloride channels share a common structure, the sites of expression in the body and their intracellular localization are different among CLCs. CLC-K1 and -K2 are kidney-specific CLC chloride channels. We have clarified their localization in the plasma membranes of tubular cells by immunohistochemistry and proposed their roles in transepithelial chloride transport. Since there exit no good inhibitors for these channels, a gene knockout approach was the only way to establish their roles in kidney. While we were generating CLC-K knockout mice, Simon et al. reported that the mutations of CLC-K2 in human resulted in Bartter's syndrome. This had been anticipated since CLC-K2 is known to be present in the basolateral plasma membranes of the distal nephron where sodium-dependent chloride transporters are present in the apical membranes. Thus, CLC-K2 constitutes an important route for chloride reabsorption as an exit for chloride ions in the basolateral membrane. Another important finding in Simon's report was that no CLC-K (a human homologue of rat CLC-K1) mutation was found in patients with Bartter's syndrome. This suggested that CLC-K1 has a different role in kidney. We believed this to be true, based on the finding that the intrarenal localization of CLC-K1 and CLC-K2 are completely different. In the CLC-K1 knockout mice, we could clearly verify that (i) the high chloride permeability in the tAL was mediated by CLC-K1 and (ii) this chloride transport is necessary for urinary concentration. Further studies are necessary to elucidate the detailed mechanisms of the urinary-concentrating defect in Clcnk1 -/- mice. Exact clearance studies and measurements of osmolality and solute contents in the inner medulla will provide the answer to this question.

Animals↗