Biomedical subjects

Y Kawai

Publications and source records attributed to Y Kawai.

At least 55 records · Page 3Linked to original sources

PubMed2001

[Reversed C.P.C. :Case of 14-year old girl with large abnormal cells in her bone marrow].

A 14-year-old female was admitted to our institute with arthralgia of the left pelvis. Magnetic resonance image analysis revealed the presence of a low-density area in her left pelvis. Although the complete blood count was almost within normal ranges, large abnormal cells with a round-shaped multi-nucleus and abundant cytoplasm containing coarse basophilic granules were noted in her bone marrow smear. She was diagnosed with mastocytosis after laboratory examinations. Nine months later, after intensive chemotherapy, the patient died.

Adolescent↗

PubMed2001

[The measurement of immature reticulocyte fractions and clinical significance].

We evaluated automated analysis for the measurement of immature reticulocyte fraction(IRF) in 222 blood samples. IRF was performed by R-3000 & XE-2100(Sysmex), VEGA RETIC LC141(Horiba), ADVIA 120(Bayer medical), CELL DYN4000(Abbott) or GEN'S(Coulter), and compared with measurements obtained by visual counting. A subfraction of IRF was also determined using different modalities on all analyzers. The values of IRF were stable after blood storage for 48 hours at 4 degrees C and 8 hours at room temperature. A good correlation of IRF among these analyzers was obtained(r = 0.629-0.869), but the normal ranges in the percentage of IRF varied according to analyzers. However, an elevation in the percentage of IRF is the first sign of bone marrow recovery after chemotherapy and the bone marrow transplantation in the majority of leukemic patients. We suggest that IRF counting is useful for the estimation of early phase erythropoiesis in patients who have successful bone marrow engraftment following bone marrow transplantation.

Hematologic Diseases↗

PubMed2001

Re-evaluation of the reticulo-endothelial system.

By injecting lithium carmine (Lit-car) into living animals, Ribbert (1904) and Kiyono (1914) showed that specific staining occurred in some cells in various organs. Kiyono termed those cells "histiocytes" which consisted of free amoeboid cells and cells of reticulo-endothelium. Aschoff (1924) introduced the concept 'reticulo-endothelial system (RES)' for the collection of cells having in common the property of vital staining. Van Furth (1972) proposed the term "mononuclear phagocyte system (MPS)" advocated that RES be replaced by MPS. As a consequence we presently suffer from a "reticulo-endothelial confusion", based on the delusion that cells other than macrophages are not members of the RES. The point that most clearly illustrates this is the fact that the Lit-car-laden cells in the body are named macrophages in modern textbooks of histology. To clarify the confusion, we re-examined the vital staining at light and electron microscopic levels and analyzed the scavenger cells using biochemical methods. Injected Lit-car was actively endocytosed by sinusoidal endothelial cells in the liver and reticular cells in the sinus of lymph nodes. Of note, uptake of the dye was comparatively much lower in macrophages/monocytes. Our findings indicate the existence of scavenger endothelial cells and reticular cells in blood and lymphatic circulations respectively. We name these two systems "the scavenger RES" collectively.

Animals↗

PubMed2000

Electrophysiological and morphological characteristics of nucleus tractus solitarii neurons projecting to the ventrolateral medulla.

Electrophysiological and morphological properties of a direct projection from the nucleus of the tractus solitarius (NTS) to the ventrolateral medulla (VLM) were investigated. NTS neurons projecting to the VLM exhibit a monosynaptic excitatory response followed by an inhibitory one after the tractus solitarius stimulation. These neurons show spontaneous inhibitory postsynaptic currents, and have medium to large soma (14-26 microm in diameter). It is concluded that the projection from the NTS to the VLM is mediated mostly by medium to large neurons that are inhibited locally by GABAergic interneurons within the NTS.

Action Potentials↗

PubMed2000

Spontaneous synaptogenesis in ex vivo sympathetic ganglion and the blockade by serum treatment.

Central denervation for more than 1 month has been shown to cause an increase in the number of adrenergic synapses in sympathetic ganglia in vivo. Here, we report several lines of evidence that adrenergic synapses may be generated de novo in ex vivo superior cervical ganglion (SCG) of adult rats only several hours after the isolation. Structures immunoreactive for synaptophysin, a marker of presynaptic elements, were drastically decreased 6 days after the preganglionic denervation. A significant increase in number of synaptophysin positive boutons was observed over 3-8 hours in the denervated SCGs maintained ex vivo at 36 degrees C in oxygenated physiologic saline, and this increase was blocked by adding normal serum in the saline. Electron microscopic analysis confirmed that the number of adrenergic synapses specifically labeled with 5-hydroxydopamine was increased by several-fold under the same condition. Intracellular labeling of SCG neurons revealed an increase in the incidence (from 8 to 50%) of neurons having dendritic plexus after the in vitro incubation. No evidence of axonal sprouting within the ganglion was observed. Intracellular recordings from single neurons of denervated SCGs revealed that maximum amplitudes of inhibitory postsynaptic potentials, which were completely blocked by yohimbine, an alpha2-adrenoceptor antagonist, in response to focal stimulation were increased over the several hours. These results suggest that dendrites of SCG neurons rapidly develop and exhibit local efferent characteristics that underlie the inhibitory synaptic transmission once they are subjected to serum deprivation.

Animals↗

PubMed2000

The molecular characterization and tissue distribution of the human cysteinyl leukotriene CysLT(2) receptor.

Cysteinyl leukotrienes (CysLTs), slow-reacting substances of anaphylaxis, are lipid mediators known to possess potent proinflammatory action. Pharmacological studies using CysLTs indicate that at least two classes of G protein-coupled receptors (GPCRs), named CysLT(1) and CysLT(2), exist; the former is sensitive and the latter is resistant to the CysLT(1) antagonists currently used to treat asthma. Although the CysLT(1) receptor gene has been recently cloned, the molecular identity of the CysLT(2) receptor has remained elusive. Here we show that the pharmacological profile of an orphan GPCR (PSEC0146) is consistent with that of the CysLT(2) receptor. In human embryonic kidney 293 cells that express the PSEC0146 cDNA, leukotriene C(4) (LTC(4)) and leukotriene D(4) (LTD(4)) induce equal increases in intracellular calcium mobilization; these increases are not affected by CysLT(1) antagonists. Additionally, [(3)H]LTC(4) specifically binds to membranes from COS-1 cells transiently transfected with PSEC0146. Large amounts of the PSEC0146 mRNA are found in human heart, placenta, spleen, and peripheral blood leukocytes but not in the lung and the trachea. Pharmacological feature and expression studies will eventually lead to a better understanding of the classification of CysLT receptors, possibly leading to a reconsideration of the pathological and physiological role of CysLTs.

Animals↗

PubMed2000

Silefrin, a sodefrin-like pheromone in the abdominal gland of the sword-tailed newt, Cynops ensicauda.

Sodefrin-like female-attracting pheromone was purified from the abdominal glands of male sword-tailed newts, Cynops ensicauda, by gel-filtration chromatography and reversed-phase high-performance liquid chromatography. The final product comprises 10 amino acid residues with the sequence SILSKDAQLK which coincided with the sequence deduced from its precursor cDNA. This peptide was designated silefrin. The sequence of silefrin was different from that of sodefrin by two amino acid residues, with substitutions Leu for Pro and Gln for Leu at positions 3 and 8, respectively. Both native and synthetic silefrin exerted an equipotent activity in attracting conspecific females.

Animals↗

PubMed2000

Involvement of Gialpha2 in sodium butyrate-induced erythroblastic differentiation of K562 cells.

The chronic myelogenous leukaemia cell line K562 can be triggered in culture to differentiate along the erythrocytic pathway in response to a variety of stimulatory agents. In the presence of sodium butyrate, these cells differentiate to erythroblasts and acquire the capability to synthesize haemoglobin. We used this cell system to study alterations in the levels of several G-protein subunits during the cell differentiation programme and to assess the involvement of G(i)alpha2 in this process. Western immunoblot analysis revealed the presence of G(s)alpha1, G(s)alpha2, G(i)alpha2, G(q)alpha, Galpha(12), Gbeta1 and Gbeta2 in K562 cells. G(o)alpha, G(z)alpha, Galpha(13) and Galpha(16) were not detected. Although the levels of several G-protein subunits were altered after treatment with sodium butyrate, the most striking change was the robust increase in the levels of G(i)alpha2, which was accompanied by an increase in the mRNA for G(i)alpha2. Inactivation of G(i)alpha2 by adding Bordetella pertussis toxin to the cultures inhibited erythroblastic differentiation by as much as 62%, as measured by haemoglobin accumulation. Furthermore, the addition of an oligonucleotide anti-sense to G(i)alpha2 inhibited the sodium butyrate-induced robust increase in G(i)alpha2 levels, decreasing it to the basal levels seen in control cells; this treatment decreased the erythroblastic differentiation of the cells (as measured by haemoglobin expression) by 50%. Taken together, these findings imply that increased levels of G(i)alpha2 contribute to the sodium butyrate-induced erythroblastic differentiation of K562 cells.

Butyrates↗

PubMed2000

Inhibition of endothelium-dependent arterial relaxation by oxidized phosphatidylcholine.

Formation of oxidized phosphatidylcholine (ox-PC), oxidatively fragmented phosphatidylcholine (PC) containing a short-chain and/or polar oxidative residue at the sn-2 position, in the process of LDL oxidation as well as its existence in atherosclerotic lesions has been demonstrated. To clarify the pathophysiological role of ox-PC in the vascular reactivity, we investigated the effects of various ox-PCs on the isometric tensions in rabbit thoracic aortas. Ox-PCs, which were produced upon oxidation of sn-2 polyunsaturated fatty acid (PUFA)-containing PCs, dose-dependently inhibited endothelium-dependent relaxation (EDR) evoked by acetylcholine or substance P. On the other hand, neither native PUFA-containing PCs nor an oxidative product of monounsaturated fatty acid-containing PC showed an inhibitory effect. None of ox-PCs affected endothelium-independent relaxation to nitroglycerin. The PC-headgroup fraction, but not the oxidized fatty acids fraction, was responsible for the inhibition of EDR by ox-PC. EDR was reduced by 2-(5-oxovaleroyl)-PC, one of the secondary oxidative products of PCs that contains a short chain aldehydic residue at the sn-2 position, but not by PC hydroperoxide, the primary oxidative product. Although the possibility could not be completely ruled out that lysophosphatidylcholine rather than ox-PC may be responsible for inhibitory effects on EDR, these results suggest a novel vascular activity of ox-PCs generated from sn-2 PUFA-containing PCs which may be implicated in the pathophysiology of vascular tone.

Acetylcholine↗

PubMed2000

The effects of binocular suture and dark rearing on the induction of c-fos protein in the rat visual cortex during and after the critical period.

It has been demonstrated in kittens that binocular lid suture has more deleterious and irreversible effects on plasticity of the developing visual system than rearing in complete darkness. The present study using immunocytochemistry focuses on the effects of the two types of visual deprivation on the inducibility of c-fos protein in visual cortical neurons of rats. Rats were subjected to binocular suture or dark rearing for 1 week during (postnatal days 14-21; P14-P21) and after (P50-P57) the critical period for activity-dependent modifiability of cortical ocular dominance. In rats of both age groups reared in the normal light-dark condition, only a small number of Fos-immunoreactive neurons was obtained in the visual cortex. By contrast, in dark-reared pups and adult rats, numerous c-fos neurons were detected in the layers II-IV and VI of the visual cortex following a brief light exposure (1 h). In rats of both ages subjected to binocular suture, Fos neurons were detected in the same layers as in the dark-reared rats, but significantly less in number. We speculate that the reduced plasticity of the visual cortex in the rats subjected to binocular suture may be due partly to the repressed AP-1 activity in visual cortical neurons. No significant difference was detected in c-fos expression in the visual cortex between visually manipulated pups and adult rats.

Aging↗

PubMed2000

Alcohol enhances characteristic releases of dopamine and serotonin in the central nucleus of the amygdala.

The amygdaloid complex (AMY) is implicated in emotional and motivational aspects of behavior, including the formation of positive reinforcement association. AMY may also associated with brain rewarding circuitry. In the present study, the effect of ethanol (EtOH) on the release of dopamine (DA) and serotonin (5-HT) was studied in the central amygdaloid nucleus (CeAMY), and projecting excitatory afferents to the ventral tegmental area (VTA), of freely moving Wistar rats by brain microdialysis. Within 20 min of i.p. injection of EtOH (2 g/kg), the levels of DA and 5-HT in the CeAMY dialysate increased over the baseline value by 270 and 160% (N = 6-7), respectively. Addition of EtOH (25, 50 and 100 mM) to the microdialysis perfusion medium for 1 h caused a 115-150% dose-related increase in the extracellular level of DA in the CeAMY. 100 mM EtOH-induced CeAMY DA release continued to increase for 1 h after the perfusion medium was returned to normal perfusion medium. In contrast, the CeAMY 5-HT level was increased only by the addition of 100 mM EtOH for 1 h to 130% for 80 min. The stimulation of the CeAMY by EtOH through the microdialysis membrane showed delayed responses of DA and 5-HT compared with the i.p. injection of EtOH. Overall, the present findings are not sufficient to conclude whether EtOH acts directly or indirectly on the major monoamine nerve cells in the CeAMY, but the degree of acute EtOH action affected the differences in time at the peak response on EtOH-induced DA and 5-HT releases in the CeAMY via VTA.

3,4-Dihydroxyphenylacetic Acid↗

PubMed2000

Postnatal differentiation of local networks in the nucleus of the tractus solitarius.

Whole-cell voltage-clamp recordings from rat brain slice preparation were used to investigate a possible developmental change in the patterns of synaptic interactions among the nucleus tractus solitarii neurons by analysing spontaneous postsynaptic current activity. Three types of patterns of spontaneous postsynaptic current activity were distinguished in the nucleus tractus solitarii neurons which showed high activities in terms of current frequency and amplitude. The first type was characterized by the presence in an individual cell of high frequencies and large amplitudes of both spontaneous glutamatergic and GABAergic postsynaptic currents, and observed exclusively in postnatal day 0-7 rats. The second and third types of cells showed predominant either inhibitory or excitatory postsynaptic currents, respectively. After postnatal day 5, nucleus tractus solitarii neurons with high background activity were shown to differentiate into either the second or the third type, with the latter of about 70% in the adult caudal/intermediate nucleus tractus solitarii. Axon collaterals of some medium to large neurons seemed to be decreased by pruning during postnatal development. The early postnatal differentiation of background synaptic activity observed in the nucleus tractus solitarii presumably reflects the local network reorganization and may be related to maturational changes in cardiovascular and respiratory functions.

Animals↗

PubMed2000

Vacuum assisted cardiopulmonary bypass in minimally invasive cardiac surgery: its feasibility and effects on hemolysis.

The present study describes a cardiopulmonary bypass (CPB) technique that incorporates vacuum assisted venous drainage and arterial return using a centrifugal pump in minimally invasive cardiac surgery (MICS). The technique was performed on 40 patients scheduled to undergo MICS. The proposed CPB technique enables a good operative field to be obtained even through a limited incision through the use of peripheral cannulation using small cannulae. Vacuum pressure was applied to the venous reservoir (-43 +/- 14 mm Hg) to maintain adequate CPB flow (>2.4 L x min-1 x M-2). The effects of CPB on hemolysis were subsequently compared between patients who underwent the proposed procedure (MICS group; n = 6) and a control group who underwent coronary arterial bypass grafting (CABG group; n = 6) with conventional CPB. Plasma free hemoglobin (FHb) increased and plasma haptoglobin (Hp) decreased during CPB in both groups, with no significant difference between the groups. By the next day, FHb had returned to pre-CPB levels whereas Hp remained lower in both groups. Again, these values did not differ significantly between groups. Thus, we conclude that the proposed CPB technique is useful in MICS with acceptable effects on hemolysis.

Analysis of Variance↗

PubMed2000

Competitive reverse transcription-polymerase chain reaction assay for quantification of human multidrug resistance 1 (MDR1) gene expression in fresh leukemic cells.

We have analyzed MDR1 gene expression in 69 clinical samples obtained from 64 patients with leukemic hematologic malignancies by using a competitive reverse transcription-polymerase chain reaction assay with a heterologous competitor RNA. To exclude a false-positive result caused by concomitant normal lymphocytes that physiologically express MDR1, in samples we determined a cut-off value of 8 amol MDR1 transcript per microgram of RNA by simultaneous measurement of rhodamine 123 dye efflux either in lymphocyte or gated leukemic cell populations. Consequently, 23 of 69 samples were concluded to be MDR1-positive in leukemic cells per se. The MDR1 expression rate was significantly correlated with factors such as a history of preceding chemotherapy, elder age of the patient, and certain disease types (eg, leukemia progressed from myelodysplastic syndrome). Moreover, the complete response rate after chemotherapy was significantly higher in MDR1-negative patients than in MDR1-positive patients (52% vs 17%, respectively; P = .01). The assay established will enable the quantification of MDR1 gene expression in blood samples from patients with leukemic hematologic malignancies and will be applicable to clinical laboratories as a routine test.

Adolescent↗

PubMed2000

Nitric oxide induces a decrease in the mitochondrial membrane potential of peripheral blood lymphocytes, especially in natural killer cells.

Increased levels of nitric oxide (NO) at an inflammatory site may affect the biological activity of lymphoid cells. To investigate the effects of NO on the immune system, we measured the mitochondrial membrane potential (delta psi m) of the peripheral blood lymphocytes (PBL) cultured with a chemical NO donor. PBL from healthy volunteers were cultured with NOC18, a NO-generating compound, at various concentrations. The delta psi m of the PBL was measured by flow-cytometry using 3,3-dihexyloxacarbocyanine iodide (DiOC6(3)). NOC18 induced a decrease in the delta psi m of the PBL in a dose-dependent fashion, induced an increase in the levels of reactive oxygen species (ROS), and caused these cells to undergo apoptosis. Dual-color staining of the delta psi m and lymphocyte surface markers demonstrated that CD3-CD56+ natural killer (NK) cells were responsive to NO. Trolox, a vitamin E analog, partially reversed the NO-induced decrease in the delta psi m of the PBL. We showed that the delta psi m of peripheral NK cells were decreased by NO, which suggests that abundant NO at an inflammatory site may impair NK cell function.

Antioxidants↗

PubMed2000

Cloning of a rat glia maturation factor-gamma (rGMFG) cDNA and expression of its mRNA and protein in rat organs.

We isolated a rat glia maturation factor-gamma(rGMFG) cDNA and examined the tissue distribution of GMFG in rat by Northern and Western blot analyses. Sequence analysis of the entire cDNA revealed an open reading frame of 426 nucleotides with a deduced protein of 142 amino acid residues. The deduced amino acid sequence of the putative product is highly homologous (78.9%) to rat glia maturation factor-beta (rGMFB). Northern blot analysis indicated that a 0.9-kb mRNA is predominantly expressed in rat thymus, testis, and spleen. GMFG showed a different tissue distribution from GMFB, being present predominantly in proliferative and differentiative organs.

Amino Acid Sequence↗

PubMed2000

Ornithine-containing lipids stimulate CD14-dependent TNF-alpha production from murine macrophage-like J774.1 and RAW 264.7 cells.

The ornithine-containing lipids (OL)-induced cytokine production pattern in macrophage-like J774.1 and RAW 264.7 cells was different from that in the peritoneal macrophages previously reported. OLs, as well as lipopolysaccharide (LPS) of Escherichia coli, strongly induced tumor necrosis factor (TNF) alpha but not interleukin (IL)-1beta in J774.1 cells. In the RAW cells, IL-1beta, TNF-alpha and prostaglandin E(2) were strongly induced by the OLs and LPS. OL- and serine-glycine-containing lipid (SGL)-induced TNF-alpha production in J774.1 and RAW 264.7 cells required serum. However, in CD14-deficient LR-9 cells, TNF-alpha was not induced by the OLs in the presence or absence of serum. OLs and a SGL almost completely inhibited the binding of (125)I-LPS to J774.1 cells. These results suggested that OLs and SGL activate macrophages via the CD14-dependent pathway.

Animals↗

PubMed2000

Modulation of chemotaxis, O(2)(-) production and myeloperoxidase release from human polymorphonuclear leukocytes by the ornithine-containing lipid and the serineglycine-containing lipid of Flavobacterium.

The ornithine-containing lipid (OL) and the serineglycine-containing lipid (SGL) of Flavobacterium activated and modulated the functions of human polymorphonuclear leukocytes (PMNs). The OL and the SGL strongly activated fMet-Leu-Phe- and interleukin-8-induced chemotaxis of PMNs at the concentration of 0.1 microg ml(-1), and a synthetic OL also activated the function of PMNs. Further, the OL strongly activated O(2)(-) production from PMNs. Although the OL and the SGL slightly modulated myeloperoxidase release from PMNs, inhibition effects of their component fatty acid analogues were observed. O(2)(-) production-inducing activity is a common biological activity between the OL and bacterial lipopolysaccharides, but OL and SGL, unlike lipopolysaccharide, are potent activators of PMN chemotaxis.

Cells, Cultured↗