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Biomedical subjects

H Uchiyama

Publications and source records attributed to H Uchiyama.

At least 19 recordsLinked to original sources

Effects of estrogenic hormones on early development of Xenopus laevis.

Many chemicals released into the environment have estrogenic activity and can disrupt animal development and the function of endocrine systems. In order to study the effects of estrogens on aquatic animals, we examined the effects of certain estrogens on early development in Xenopus laevis. X. laevis embryos were kept in water containing 10(-10), 10(-9), 10(-7), 10(-6), and 10(-5) M 17 beta-estradiol (E2); 17 alpha-estradiol; diethylstilbestrol (DES); 10(-5) M progesterone (P); or dihydrotestosterone (DHT) beginning at developmental stage 3. Survival rates of the embryos developed in water containing 10(-10)-10(-6) M E2 or DES, all concentrations of 17 alpha-estradiol, and 10(-5) M P or DHT, which were over 70% after stage 48, whereas the rates of the embryos treated with 10(-5) M E2 and DES decreased remarkably after stage 27 and all embryos were dead by stages 42 and 32, respectively. Embryos treated with 10(-5) M E2 showed malformations of the head and abdomen and suppressed organogenesis, including crooked vertebrae at stage 38; the head was smaller and the abdomen was larger than in the controls. Similar effects were observed in embryos developed in 10(-5) M DES but not in 10(-5) M 17 alpha-estradiol, P, or DHT. After 10(-5) M E2 treatment, abnormalities were induced only when the treatment was started before stage 39. However, on day 30 after fertilization, the stage of the embryos treated with 10(-6) M E2 was more progressed than that of the controls. Estrogen receptor (ER 4) mRNA was examined in eggs, embryos, and adult female liver by reverse-transcription polymerase chain reaction. ER4 mRNA was expressed in adult liver, unfertilized and fertilized eggs, and embryos, but ER3 mRNA was not expressed. ER4 mRNA in 10(-6) and 10(-5) M E2-treated embryos showed different expression patterns, which may result from the diverse developmental effects of E2. The present results demonstrate that 10(-5) M E2 and DES induced embryo death and malformations and that ER may be involved in the induction of various developmental defects in X. laevis embryos.

Abdomen

Phase Behavior of a Water/Nonionic Surfactant/Oil Ternary System in the Presence of Polymer Oil

The effect of a polymer oil, polydimethyl siloxane, on the phase behavior of the water/C12EO6/isopropyl myristate (IPM) system has been studied. Since the polymer oil is completely soluble in IPM, it was dissolved in IPM and the solutions were used as the oil phase to study its effect. The presence of polymer increases the hydrophobic nature of IPM and thereby decreases the solubilization of oil into the surfactant phase (microemulsion). Moreover, at a certain range of silicone oil concentration in the IPM, a four-phase body consisting of excess water, excess oil, and two surfactant phases is formed within the ternary system. The two surfactant phases are designated as D (with bicontinuous type of structure) and D' (with L3 type of structure). Careful phase behavioral studies revealed that, with increasing silicone oil concentration, a three-phase region containing water phase, D' phase, and an oil-rich D phase develops within the system and this region overlaps with the normal three-phase region of the system containing the water, D, and oil phases to give a four-phase body. The order of various phases from top to bottom in the four-phase body is oil, D, D', and water.

Journal Article

Serum thrombopoietin levels in patients undergoing autologous peripheral blood stem cell transplantation.

Recently, the ligand for c-mpl has been cloned and initial studies have shown it to be the platelet regulatory factor, thrombopoietin (TPO). To elucidate the role of TPO in the reconstitution of megakaryopoiesis and platelet production after stem cell transplantation, we measured serum TPO levels in nine patients undergoing autologous peripheral blood stem cell transplantation (PBSCT) and in healthy volunteers. Serum TPO levels significantly correlated with the degree of peripheral thrombocytopenia and a strong inverse correlation between serum TPO level and platelet count was observed (r = -0.700, P < 0.001). Serum TPO levels began to rise as the platelet count decreased after chemotherapy, TPO levels peaked at over 25.00 fmoles/ml between days 0 and 10; TPO levels then decreased gradually as the platelet count began to rise. One patient with multiple myeloma received purified CD34+ peripheral blood stem cells. No difference was observed in the kinetics of serum TPO levels between unfractionated and purified PBSCT. These observations suggest that TPO plays a critical role in the reconstitution of megakaryopoiesis and platelet production after PBSCT.

Adolescent

The soluble methane monooxygenase gene cluster of the trichloroethylene-degrading methanotroph Methylocystis sp. strain M.

In methanotrophic bacteria, methane is oxidized to methanol by the enzyme methane monooxygenase (MMO). The soluble MMO enzyme complex from Methylocystis sp. strain M also oxidizes a wide range of aliphatic and aromatic compounds, including trichloroethylene. In this study, heterologous DNA probes from the type II methanotroph Methylosinus trichosporium OB3b were used to isolate souble MMO (sMMO) genes from the type II methanotroph Methylocystis sp. strain M. sMMO genes from strain M are clustered on the chromosome and show a high degree of identity with the corresponding genes from Methylosinus trichosporium OB3b. Sequencing and phylogenetic analysis of the 16S rRNA gene from Methylocystis sp. strain M have confirmed that it is most closely related to the type II methanotroph Methylocystis parvus OBBP, which, unlike Methylocystis sp. strain M, does not possess an sMMO. A similar phylogenetic analysis using the pmoA gene, which encodes the 27-kDa polypeptide of the particulate MMO, also places Methylocystis sp. strain M firmly in the genus Methylocystis. This is the first report of isolation and characterization of methane oxidation genes from methanotrophs of the genus Methylocystis.

Amino Acid Sequence

Characteristics of Escherichia coli HB101 and Pseudomonas putida PpY101 harboring a recombinant plasmid with tandem insertion of the mercury resistance operon.

We constructed the plasmid pSUPmer2 by inserting tandem copies of the mercury resistance (mer) operon into a broad host range-vector, and introduced it into Escherichia coli HB101 and Pseudomonas putida PpY101 to increase their mercury resistance. Strains harboring plasmid pSUPmer2 had higher mercury resistance and mercuric reductase activity than those strains harboring the plasmid pSUPmer which had one copy of the mer operon. Mercury resistance of P. putida PpY101 was significantly increased by tandem insertion of the mer operon.

Cell Division

Fusion with postpaced return cycle identical to tachycardia cycle length during transient entrainment of ventricular tachycardia and its implications.

In reentrant ventricular tachycardia (VT), the postpaced return cycle (RC) during transient entrainment at a pacing site far from the central common pathway is longer than the VT cycle length (VTCL), when VT is represented by a figure-eight model. However, the reentrant circuit has not been fully elucidated. The purpose of this study was to present VT in which the postpaced RC became identical to VTCL during transient entrainment while fusion is evident in the surface electrocardiogram (ECG). Among 38 patients with inducible reentrant VTs who underwent electrophysiologic study (EPS), 10 VTs of six patients were selected. All patients had underlying heart diseases: dilated cardiomyopathy (n = 2), coronary artery disease (n = 1), postoperative tetralogy of Fallot (TOF; n = 2), and arrhythmogenic right ventricular dysplasia (n = 1). Catheter mapping was performed to demonstrate that the site of origin was distant from the pacing site. The cycle length of induced VT (n = 10 VTs) was 380 +/- 41 msec. Five patients (83%) had two morphological VTs; one a left bundle branch block (LBBB) and the other a right bundle branch block (RBBB) pattern. During rapid pacing, constant fusion was observed in all VTs, but the postpaced RC was identical to VTCL. In 2 patients (4 VTs), the revolution of wavefronts around an anatomical obstacle (scar of myotomy in TOF, and infarction) was demonstrated. The fact that the postpaced RC was identical to VTCL but showed fusion in the surface ECG can be explained by macro-reentry. The pacing site must be located at the preferential route of the macroreentrant circuit.

Adolescent

Effectiveness of aldose reductase inhibitors for diabetic gastroenteropathy with constipation.

We present a diabetic patient with long-standing constipation complicated by paralytic ileus and septic shock. She successfully recovered from a critical condition, and her diabetes was well controlled. However, the constipation did not improve even after the administration of conventional medications. Epalrestat, an aldose reductase inhibitor (ARI), improved her bowel motility and autonomic cardiovascular dysfunction, as evident from her heart rate and blood pressure response. Gastroenteropathy is a major diabetic complication which may cause disturbed bowel motility leading to serious enterobacterial infections, thus, its amelioration is important. ARI may be beneficial in the treatment of diabetic gastroenteropathy refractory to conventional therapies.

Aldehyde Reductase

[Basilar artery aneurysm associated with agenesis of unilateral internal carotid artery: two case reports].

Two patients presented with a basilar artery aneurysm associated with agenesis of the right internal carotid artery. In both cases, the right middle cerebral artery was supplied by the basilar artery via the enlarged right posterior communicating artery, and the right anterior cerebral artery was supplied by the left internal carotid artery via the anterior communicating artery. The first patient was a 65-year-old woman who suffered from a subarachnoid hemorrhage due to rupture of a basilar bifurcation aneurysm, which was demonstrated as a "de novo" aneurysm 13 years after successful clipping of the anterior communicating artery aneurysm. Delayed surgery was planned, but she died due to recurrent hemorrhage. The second patient was a 67-year-old woman who had had a history of progressive left sided weakness over the previous few years. Conventional angiography and 3 dimensional CT angiography showed a large aneurysm of the basilar artery trunk with a wide neck. Endovascular embolization was performed with mechanically detachable coils, and the aneurysm was incompletely occluded. The patient regained complete strength in the left arm after the treatment, but the follow-up angiography at 5 months disclosed recanalization of the treated aneurysm, associated with shift of the packed coils. Hemodynamic stress resulting from unique collateral circulation with agenesis of the internal carotid artery may cause a predisposition to "de novo" aneurysm formation or recanalization of an occluded aneurysm with coils.

Aged

Migration of GnRH-immunoreactive neurons from the olfactory placode to the brain: a study using avian embryonic chimeras.

Previous studies suggest that gonadotropin-releasing hormone (GnRH) neurons appear in the olfactory placode and subsequently migrate into the brain during embryonic development. The aim of the present study was to obtain direct evidence for migration of GnRH neurons from the olfactory placode into the brain. Olfactory placodes from quail embryos were transplanted isotopically and isochronically, to replace the unilaterally ablated olfactory placodes of chick embryos. The chimeric embryos were allowed to develop for several days until they reached the embryonic stages when GnRH neurons are seen in the brain in normal embryos. Quail olfactory epithelia were formed in the host chick embryos. Quail olfactory nerves were also formed and reached the olfactory bulb or primordial olfactory bulb. GnRH-immunoreactive cells of quail origin revealed by a triple staining method were observed in the quail olfactory epithelium, quail olfactory nerve, chick olfactory bulb, and septo-preoptic area. These results indicate that GnRH neurons originate in the olfactory placode and migrate into the telencephalon including the septo-preoptic area. A migratory route of GnRH neurons was well documented by the use of a quail neuron-specific antibody, QN. The migratory route in the brain is discussed with special reference to the terminal nerve. A GnRH-immunoreactive neuronal group of chick origin appeared in the diencephalon of chimeric embryos. These diencephalic neurons may be of non-placodal origin. FMRFamide-immunoreactive neurons of quail origin were also found in the quail olfactory nerve and the host olfactory bulb, suggesting that FMRFamide neurons also originate in the olfactory placode and migrate into the brain.

Animals

Phosphorylation of human m1 muscarinic acetylcholine receptors by G protein-coupled receptor kinase 2 and protein kinase C.

Human muscarinic acetylcholine receptor m1 subtypes (m1 receptors) were expressed in and purified from insect Sf9 cells and then subjected to phosphorylation by G protein-coupled receptor kinase 2 (GRK2) expressed in and purified from Sf9 cells and by protein kinase C purified from rat brain (a mixture of alpha, beta, and gamma types, PKC). The m1 receptor was phosphorylated by either GRK2 or PKC in an agonist-dependent or independent manner, respectively. G protein beta gamma subunits stimulated the phosphorylation by GRK2 but did not affect the phosphorylation by PKC. The number of incorporated phosphates was 4.6 and 2.8 mol/mol of receptor for phoshorylation by GRK2 and PKC, respectively. The number of incorporated phosphates was 7.5 mol/mol receptor for phosphorylation by GRK2 followed by PKC, but was 5.8 mol/mol of receptor for the phosphorylation by PKC followed by GRK2. Major sites phosphorylated by GRK2 and PKC were located in the third intracellular loop and the carboxyl-terminal tail, respectively. These results indicate that GRK2 and PKC phosphorylate different sites of m1 receptors and that the phosphorylation by PKC partially inhibits the phosphorylation by GRK2, probably by affecting activation of GRK2 by agonist-bound receptors.

Amino Acid Sequence

Multiple myeloma cell adhesion-induced interleukin-6 expression in bone marrow stromal cells involves activation of NF-kappa B.

Adhesion of multiple myeloma (MM) cells to bone marrow stromal cells (BMSCs) not only localizes MM cells in the marrow microenvironment, but also triggers interleukin-6 (IL-6) secretion by BMSCs and related MM cell proliferation. In the present study, we characterized the regulation of IL-6 gene expression in BMSCs during MM cell adhesion. Adhesion of ARH-77, HS-Sultan, IM-9, and U266 MM cell lines to BMSCs and BMSC lines (LP 101 and AA 101) triggered 5-through 15-fold and 2-through 4-fold increases in IL-6 secretion, respectively. IL-6 mRNA transcripts were undetectable by Northern blotting in IM-9 MM cells or LP 101 BMSCs cultured alone; however, adherence of IM-9 cells to LP 101 cells induced a transient increase in IL-6 transcripts at 6 hours, followed by peak IL-6 secretion at 24 hours. To confirm increased IL-6 transcription and characterize its regulation, LP101 BMSCs were transiently transfected with full length and deletion fragments of the IL-6 promoter linked to the chloramphenicol acetyltransferase (CAT) reporter gene. Transient transfection of LP101 BMSCs with plasmid containing an intact NF-kappa B site showed a 6.8 +/- 0.4-fold increase in CAT activity triggered by IM-9 MM cell adhesion (n = 3, P < .05). Transfection of LP 101 cells with plasmid containing a single base pair deletion from the NF-kapp B binding motif abolished the MM adhesion-induced increase in CAT activity, whereas transfection with plasmid containing three copies of synthetic NF-kappa B sequence resulted in an 8.1 +/- 0.7-fold increase in CAT activity related to MM adhesion (n = 3, P < .05). These data suggest that the NF-kappa B site is one of the essential regulatory elements for MM cell adhesion-induced IL-6 transcription in BMSCs. Electrophoretic mobility shift assays confirmed the involvement of NF-kappa B activation in regulating MM adhesion-induced IL-6 transcription in BMSCs. Further characterization of the upstream events in the signalling cascade regulating IL-6 may not only delineate mechanisms of IL-6 regulation during paracrine MM cell growth, but also provide new therapeutic strategies based on interruption of IL-6 mediated tumor cell growth.

Base Sequence

Detection of undegraded oligonucleotides in vivo by fluorescence resonance energy transfer. Nuclease activities in living sea urchin eggs.

A method was investigated for monitoring the integrity of oligonucleotides in solution and in cells using fluorescence resonance energy transfer between two different fluorochromes attached to a single oligonucleotide. Ten-mer oligodeoxyribonucleotides labeled with fluorescein at one end and with rhodamine X at the other end were used. The oligomer had a specific absorption spectrum with peaks at 497 and 586 nm, which corresponded to fluorescein and rhodamine X, respectively. When excited at 494 nm, the oligomer had a specific fluorescence spectrum with peaks at 523 and 610 nm. The fluorescence intensity at 610 nm was 6-8 times higher than that at 523 nm. After digestion of the oligomer with an endonuclease, the fluorescence at 523 nm increased more than 12-15-fold but its fluorescence peak at 610 nm almost completely disappeared. To examine effects in vivo, sea urchin eggs were injected with a solution of the oligomer and excited with blue light at 470-490 nm. Two fluorescent images, a green image at 520-560 nm and a red image at above 580 nm, were obtained when a single egg was viewed under a fluorescence microscope. The ratio of the intensities of red to green fluorescence decreased in dependence on time after injection of the oligomer. These changes were not observed in eggs that had been injected with a solution of similarly double-labeled, phosphorothioate oligomer. These results indicated that unfertilized sea urchin eggs had nucleolytic activity. Analysis in vitro on supernatant of the egg homogenate indeed demonstrated the existence of nucleases. All together, our results indicate that the integrity of oligonucleotides can be estimated in living cells by monitoring the fluorescence resonance energy transfer of the double-labeled oligonucleotide.

Animals

Occurrence of immunoreactive activin/inhibin beta(B) in gonadotrophs, thyrotrophs, and somatotrophs of the Xenopus pituitary.

An antibody against the Xenopus activin/inhibin beta(B) subunit (94-107) was raised in a rabbit. Using this antibody, the distribution of activin/inhibin beta(B) immunoreactivity in the pituitary of adult X. laevis was studied. Beta(B) immunoreactivity was detected in gonadotrophs, thyrotrophs, and somatotrophs under light microscopy. Electron microscopy revealed that a beta(B)-immunoreactive substance exists in LH, TSH, and GH granules, in contrast to findings in the rat and goldfish. These results indicate that the expression of activin/inhibin beta(B) in pituitary cells is not consistent among vertebrate species.

Activins

Tuberculosis following liver transplantation: report of a case and review of the literature.

We report on a 44-year-old man who developed tuberculosis 4 months after liver transplantation. The diagnosis was confirmed using a polymerase chain reaction (PCR) technique in bronchial alveolar lavage (BAL) fluid, and the patient was successfully treated by reducing his immunosuppression and administering antituberculous drugs. The patient became afebrile 20 days after starting antituberculous therapy and remains well at home. A review of the literature revealed that tuberculosis after liver transplantation is a rare complication with a reported mortality rate of as high as 40%. The mortality is highest for patients who become symptomatic within 3 months after transplantation (83% vs 0%, P < 0.01; Fisher's exact test) and for those with an interval between the initial symptom and diagnosis of more than 2 weeks (71% vs 0%, P < 0.05). Early diagnosis is, therefore, essential for successful resolution of tuberculosis after liver transplantation.

Adult

Testicular morphological changes in children with acute lymphoblastic leukemia following chemotherapy.

Morphological changes in the testis induced by chemotherapy given according to the Tokyo Children's Cancer Study Group (TCCSG) regimens were studied in children with acute lymphoblastic leukemia (ALL). After informed consent, testicular biopsies were performed 14 times in 12 patients at the end of treatment. The testicular morphology in all cases had sustained a degree of damage. The tubular fertility index (TFI), calculated as the percentage of seminiferous tubules containing identifiable spermatogonia, was from 0 to 42.8% (mean 33.4%) below the normal value. Infiltration of leukemic cells was the most significant factor contributing to the decrease in TFI. There were no differences in the TFI among the TCCSG protocols. Formation of sperm was recognized in six cases, whose ages were 7, 8, 9, 10, 15 and 19 years. In two children, testicular biopsy was performed twice. In the second biopsy, TFI was elevated and sperm formation with the maturation of Leydig cells was observed. A number of other pathological changes were observed: modification of spermatogonia, Sertoli cells and inclusion bodies in spermatogonia, abnormal maturation of Leydig cells, evidence of interstitial fibrosis and thickening of the basement membrane. These results suggest that recent strong chemotherapy for the treatment of ALL might cause severe but not fatal damage to children's testicular tissue. As chemotherapy escalates, more investigation of testicular function will be necessary.

Antineoplastic Combined Chemotherapy Protocols

Two cases of ventricular parasystole associated with ventricular tachycardia.

In two patients, ventricular parasystole (VP) was associated with ventricular tachycardia (VT), and in one patient, catheter ablation was successful. In patient 1, with dilated cardiomyopathy, VP led to VT, which converted to ventricular fibrillation. In patient 2, VP led to symptomatic nonsustained polymorphic VT. The origin of parasystolic focus was determined by endocardial mapping, and a radiofrequency current was delivered to patient 2. Both VP and VT disappeared immediately, and no recurrence has been observed during a follow-up of 8 months. Catheter ablation to the parasystolic focus was effective and a relationship between VP and VT was strongly suggested.

Cardiomyopathy, Dilated