Biomedical subjects

H Gotoh

Publications and source records attributed to H Gotoh.

At least 37 records · Page 2Linked to original sources

PubMed1999

Augmented proliferation of human alveolar macrophages after allogeneic bone marrow transplantation.

After allogeneic bone marrow transplantation (allo-BMT), recipient alveolar macrophages (AM) are gradually replaced by AM of the donor origin. An influx of mononuclear phagocytes of donor origin to the lung is responsible for the repopulation, but the detailed kinetics remain unclear. We therefore studied 24 BMT recipients who underwent bronchoalveolar lavage (BAL) from 24 to 83 days after BMT. AM cell number, size, morphology, proliferating ability, and genotype of AM were measured. Before day 50, the number and size of AM in BAL fluid were similar to those of normal nonsmokers. However, after day 50, the mean number of AM increased threefold and the mean cell size decreased due to the increase of small AM. These small cells are presumably of donor origin based on DNA fingerprinting analysis and based on fluorescence in situ hybridization for the Y chromosome in a sex-mismatched case. Immunohistochemistry and cell cycle analysis demonstrated that the increase in AM number coincided with a remarkable increase of AM expressing proliferating cell nuclear antigen, suggesting that small AM are proliferating. This is the first report representing that augmented proliferation of donor AM in situ may contribute to the reconstitution of AM population after BMT.

Adolescent↗

PubMed1999

Diagnosis of fetal pulmonary hypoplasia by measurement of blood flow velocity waveforms of pulmonary arteries with Doppler ultrasonography.

OBJECTIVE: The aim of the study was to determine the utility of ultrasonographically recorded blood flow waveforms of the pulmonary artery in the diagnosis of pulmonary hypoplasia. STUDY DESIGN: The normal values of the pulsatility index and peak systolic flow of pulmonary arterial blood velocity waveforms were determined in 300 uncomplicated single fetus pregnancies with well-established gestational ages between 24 and 40 weeks. We also measured the same parameters in 5 fetuses at high risk for development of severe pulmonary hypoplasia. We also determined the radial alveolar count and microvessel density, representing the extent of angiogenesis, in tissue specimens collected at autopsy from normal and hypoplastic lungs and stained both with hematoxylin and eosin and immunohistochemically for factor VIII. RESULTS: In healthy fetuses the pulsatility index values of both the right and left pulmonary arteries diminished significantly with advancing gestation, whereas the peak systolic velocity increased significantly with advancing gestation. In fetuses with pulmonary hypoplasia pulsatility index values were high and the peak systolic flow was significantly lower than in healthy fetuses. Histologic examination showed a lower radial alveolar count and poorer angiogenesis in fetuses with pulmonary hypoplasia than in healthy fetuses. CONCLUSION: Doppler ultrasonographic determination of pulmonary artery blood velocity waveforms is a useful tool for the diagnosis of pulmonary hypoplasia.

Blood Flow Velocity↗

PubMed1999

Mechanical stimuli induce intracellular calcium response in a subpopulation of cultured rat sensory neurons.

Cultured dorsal root ganglion neurons from newborn rats were mechanically deformed with a fine-tipped glass capillary, and the change in the intracellular Ca2+ concentration ([Ca2+]i) was recorded by Fura-2-based microfluorimetry. The deformation evoked elevation in [Ca2+]i from 18.7 +/- 5.4 nM (mean +/- S.E.M., n = 35) to 137.1 +/- 15.2 nM in some subpopulations of cells, especially those larger than 20 microm in diameter. The largest mechanosensitive cell group was that of cells 20-25 microm in diameter; 56% of the mechanosensitive cells were of this cell size. All of the cells larger than 25 microm in diameter displayed the Ca2+ increase when prodded. The depletion of extracellular Ca2+ diminished the Ca2+ elevation. Verapamil and nickel, blockers of voltage-dependent Ca2+ channels, did not influence the Ca2+ response, whereas gadolinium, a relatively selective blocker of mechanosensitive channels, diminished the response. Na+-free conditions did not influence the response. We concluded that the mechanical stimulation induced a Ca2+ influx in large dorsal root ganglion neurons through mechanosensitive Ca2+-permeable channels.

Animals↗

PubMed1999

Calcium signaling in cold cells studied in cultured dorsal root ganglion neurons.

Thermosensitive cold cells were identified in cultured dorsal root ganglion neurons from newborn rats. The neurons were loaded with a calcium indicator, Fura-PE3, and the change in intracellular Ca2+ concentration ([Ca2+]i) of the neurons was measured with microfluorimetry. Thirteen per cent of the cells responded to the cold stimulation. The diameter of the responder cells was 16.3+/-3.2 microm (mean+/-S.D., n = 25). The lowering of the temperature from 35 degrees C to 20 degrees C increased [Ca2+]i from 59.6+/-10.6 nM to 203.4+/-14.8 nM (n = 25). The [Ca2+]i response was dependent on the intensity of the cold stimulation. The depletion of extracellular Ca2+ diminished the Ca2+ elevation. However, a Na(+)-free condition did not influence the response. We concluded that the cold stimulation opens Ca2(+)-permeable channels in putative cold cells from dorsal root ganglion neurons.

Animals↗

PubMed1998

Male-specific transcription initiation of the C4-Slp gene in mouse liver follows activation of STAT5.

The mouse genes encoding the constitutively expressed complement component C4 and its closely related isoform C4-Slp (sex-limited protein), which is expressed only in male animals of several strains, provide a unique model to study sequence elements and trans-acting factors responsible for androgen responsiveness. Our previous studies have shown that hormonal induction of C4-Slp is mediated by a sex-specific pattern of growth hormone secretion. Promoter analyses in vitro have led to contradictory conclusions concerning the significance of C4-Slp-specific sequences in the 5' flanking region. Mutant mice carrying the H-2(aw18) haplotype, which is characterized by a large deletion in the S region covering the C4 and 21-OHase A genes, permit the direct in vivo analysis of C4-Slp transcription, unhindered by the presence of C4. Run-on analysis of transcription in liver nuclei of males and females of this strain demonstrated a 100-fold higher transcriptional activity in males, essentially determined at the transcription initiation level. The androgen dependence of transcription initiation was confirmed by run-on analysis of testosterone-treated females, where transcriptional activity started after 6 days of androgen treatment and reached male levels after 20 days. Conversely, the growth hormone-regulated activity of transcription factor STAT5 was already detected in liver nuclei after 48 hr of androgen treatment. Furthermore, we demonstrate that activated STAT5 recognizes in vitro two upstream gamma interferon-activated sequence (GAS) elements of the C4-Slp gene, centered at positions -1969 and -1831. We postulate that binding of STAT5 to these C4-Slp-specific GAS elements plays a crucial role in the chromatin remodelings that lead to transcriptional competence of the C4-Slp gene in the liver.

Animals↗

PubMed1998

Warm cells revealed by microfluorimetry of Ca2+ in cultured dorsal root ganglion neurons.

Warm cells were identified by Fura-PE3-based microfluorimetry of Ca2+ in cultured dorsal root ganglion (DRG) neurons. In response to a physiologically relevant stimulus temperature (43 degrees C), a subpopulation of small DRG neurons from new born rats increased the intracellular Ca2+ concentration ([Ca2+]i). Seven percent of the cells responded to the warm stimulus. The stimulus evoked elevation in [Ca2+]i from 52.5 +/- 9.5 nM (mean +/- S.D., n = 18) to 171.0 +/- 15.6 nM in cells between 15 and 25 microns in diameter. The depletion of extracellular Ca2+ diminished the Ca2+ elevation. The Na(+)-free condition also diminished the response. We concluded that the heat stimulation opens nonselective cation channels in putative warm cells from DRG neurons.

Animals↗

PubMed1998

Cloning and characterization of two human cDNAs encoding the mRNA capping enzyme.

Previous studies demonstrated that the mammalian mRNA capping enzyme is a bifunctional enzyme containing RNA 5'-triphosphatase and mRNA guanylyl-transferase activities in a single polypeptide. In yeast, both the above activities are separated into two different subunits, alpha and beta, the genes for which we have cloned recently. It is thus interesting to compare the structural and functional relationships between the mammalian and yeast capping enzymes. Here we isolated two human cDNAs encoding mRNA capping enzymes termed hCAP1a and hCAP1b which encode 597 and 541 amino acids, respectively. They are different only at the region coding for the C-terminal portion of the enzyme. Comparison of the deduced amino acid sequences with other cellular and viral capping enzymes showed that all the regions conserved among mRNA guanylyltransferases are observed in our clones except one conserved C-terminal region which was absent in the hCAP1b protein. The purified recombinant hCAP1a gene product, hCAP1a, exhibited both RNA 5'-triphosphatase and mRNA guanylyltransferase activities. Deletion mutant analysis of hCAP1a showed that the N-terminal 213 amino acid fragment containing a tyrosine specific protein phosphatase motif catalyzed the RNA 5'-triphosphatase activity and the C-terminal 369 amino acid fragment exhibited the mRNA guanylyltransferase activity. On the other hand, hCAP1b showed RNA 5'-triphosphatase activity, but neither enzyme-GMP covalent complex formation nor cap structure formation was detected.

Amino Acid Sequence↗

PubMed1998

Depolarization triggers intracellular magnesium surge in cultured dorsal root ganglion neurons.

Intracellular magnesium concentration ([Mg2+]i) of cultured dorsal root ganglion (DRG) neurons was measured using the magnesium indicator Mag-Fura-2/AM. [Mg2+]i was 0.48 +/- 0.08 mM (mean +/- SEM, n = 23) at rest, and it increased 3-fold by depolarization with a 60-mM K+ solution. The [Mg2+]i increase was observed in the absence of extracellular Mg2+, but the increase disappeared in the absence of extracellular Ca2+. 50 microM cadmium or 100 microM verapamil, a Ca2+ channel blocker, also diminished the rise of [Mg2+]i. The additional measurement of an intracellular Ca2+ concentration ([Ca2+]i) indicated that the [Mg2+]i rise requires a threshold concentration of [Ca2+]i to be reached; above 60 nM. The present results indicate that depolarization induces a Ca2+-influx through voltage dependent Ca channels and this causes the release of Mg2+ from intracellular stores into the cytoplasm.

Animals↗

PubMed1998

Mouse spermatogenic cell-specific type 1 hexokinase (mHk1-s) transcripts are expressed by alternative splicing from the mHk1 gene and the HK1-S protein is localized mainly in the sperm tail.

Unique type 1 hexokinase (HK1) mRNAs are present in mouse spermatogenic cells (mHk1-s). They encode a spermatogenic cell-specific sequence region (SSR) but not the porin-binding domain (PBD) necessary for HK1 binding to porin on the outer mitochondrial membrane. This study determined the origin of the multiple Hk1-s transcripts in mouse spermatogenic cells and verified that they are translated in mouse spermatogenic cells. It also showed that a single mHk1 gene encodes the mHk1 transcripts of somatic cells and the mHk1-sa and mHk1-sb transcripts of spermatogenic cells, that alternative exons are used during mHk1 gene expression in mouse spermatogenic cells, and that mHK1-S is translated in mouse spermatogenic cells and is localized mainly with the fibrous sheath in the tail region, not with the mitochondria in the midpiece of mouse sperm.

Alternative Splicing↗

PubMed1998

Effect of liposome-encapsulated hemoglobin, neo red cells, on hemorrhagic shock.

We examined the effects of liposome-encapsulated hemoglobin, neo red cells (NRCs), on hemorrhagic shock in a canine model. The dogs were divided into the three groups according to treatment. In group 1, composed of six dogs, NRCs were substituted for blood without shock being induced; in group 2, composed of six dogs, NRCs were administered immediately after mild shock had been induced by exsanguination through the vein; and in group 3, composed of seven dogs, NRCs were administered after they had been left untreated for 30 min inducing severe shock. In group 2, administration of NRCs at a dose equivalent to the volume of exsanguinated blood improved the symptoms of shock; however, in group 3, a dose of NRCs 1.6-times the volume of exsanguinated blood was required. Peripheral vascular resistance (PVR) decreased after NRC administration in groups 1 and 2, but increased in group 3. On the other hand, the cardiac index (CI) increased in groups 1 and 2, and decreased in group 3. Concerning oxygen kinetics, there were no increases in the oxygen requirements or arteriovenous differences of the oxygen content per hemoglobin (AV/Hb) for NRCs in groups 1 and 2. Conversely, in group 3, the oxygen requirements increased and the NRCs compensated for the decrease in CI with an increase in AV/Hb by enhancing the oxygen transport efficiency to cope with the increased oxygen requirements.

Animals↗

PubMed1998

Fetal blood flow redistribution in term intrauterine growth retardation (IUGR) and post-natal growth.

OBJECTIVE: The purpose of the present study was to evaluate the relationship between intrauterine growth retardation, represented by fetal blood flow redistribution and infantile growth. METHODS: The blood flow velocity waveform of umbilical and middle cerebral arteries was recorded by Doppler ultrasonography in 77 growth-retarded fetuses. We recorded the pulsatility index (PI) of the umbilical artery and middle cerebral and calculated the PI ratio of the middle cerebral to umbilical artery (C/U ratio). We selected a cutoff value of 1.0 to distinguish between normal and abnormal C/U ratios. To evaluate the development of infants, height, body weight and head circumference were measured serially at 3, 6 and 12 months of age. RESULTS: Birth weight was significantly lower in newborns who had abnormal C/U ratios (2021+/-295 g vs. 2294+/-236 g). The number of cases admitted to NICU was higher and the duration of admission was longer in abnormal C/U ratio group (15/29 and 33.7+/-13.5 days) than normal C/U ratio group (7/48 and 26.6+/-5.6 days). Body weight was lower in infants of abnormal C/U ratio group at 3 months, but was not different at 6 and 12 months. The height was shorter in abnormal C/U ratio group at birth, there were no significant differences at 3, 6 and 12 months. The number of newborn infants with growth retardation was slightly higher in abnormal C/U ratio group, but the difference was not significant at 6 and 12 months. There was no relationship between C/U and growth variables recorded at birth and follow-up in infants. CONCLUSION: Intrauterine growth-retardation, represented by abnormal fetal blood redistribution was associated with adverse perinatal outcome, but the influence was not found at the first year of life.

Adult↗

PubMed1998

Detection and assessment of pyelectasis in the fetus: relationship to postnatal renal function.

OBJECTIVE: To assess the relationship between fetal pyelectasis determined by ultrasonography and postnatal renal function. METHODS: We first established normal values for anteroposterior, transverse, and longitudinal diameters of the renal pelvis sonographically in 68 normal fetuses at 30-40 weeks' gestation. We also evaluated prospectively the relationship between the severity of pyelectasis (diameter at least two standard deviations above the normal mean) in 36 fetuses at 30-40 weeks' gestation and postnatal renal function. Fetal renal function also was evaluated through measurement of the hourly urine production rate, whereas postnatal renal function was assessed by technetium 99m-diethylenetriamine penta-acetic acid renography and excretory urography. RESULTS: Fetal pyelectasis was diagnosed when renal pelvic diameters were at least 8 mm (anteroposterior), 11 mm (transverse), and 14 mm (longitudinal), all upper limits of normal diameters of 68 normal fetuses as determined in the first part of the study. Pyelectasis was associated with a normal urine production rate in all fetuses, but in six fetuses it was associated with a normal urine production rate in all fetuses, but in six fetuses it was associated with a progressive deterioration of renal function within the 1st month of life, requiring neonatal surgery. In the other 30 fetuses with pyelectasis, who did not require neonatal surgery, pyelectasis decreased gradually or totally disappeared within 2 years. Renal pelvic anteroposterior, transverse, and longitudinal pelvic diameters were at least 20, 25, and 26 mm, respectively, during late fetal life in those neonates at the time of corrective surgery. The mean anteroposterior diameter in those fetuses who did not require surgery at infancy (11 +/- 6 mm) was significantly less than in those requiring surgery (33 +/- 14 mm, P < .01). Likewise, the transverse diameters were 17 +/- 10 mm and 38 +/- 16 mm (P < .01) and the longitudinal diameters were 20 +/- 10 mm and 48 +/- 22 mm (P < .01). CONCLUSION: Neonatal surgery is recommended when the anteroposterior, transverse, and longitudinal renal pelvic diameters during the prenatal period are at least 20, 25, and 26 mm, respectively. Surgery is not necessary when the diameters are less than 20 mm.

Dilatation, Pathologic↗

PubMed1998

Effect of experimentally induced urethral obstruction and surgical decompression in utero on renal development and function in rabbits.

To investigate the effect of urethral obstruction during late fetal life on renal development and function, we developed a rabbit fetal model of obstructive nephropathy to examine the pathological and biochemical consequences of urethral obstruction and beneficial effects of early surgical decompression. Animals were divided into four groups, i.e., obstructed, early decompressed, late decompressed, and control. Fetal renal development was evaluated by histological examination and counting the number of glomeruli in the four groups. The number of renal glomeruli correlated with gestational age in the normal fetus (r = 0.90, P < 0.0001). Urethral ligation on gestational day 25 (full-term, 31 days) resulted in thinning of the renal cortex and significantly decreased the number of renal glomeruli. The concentration of urinary microalbumin was higher when urethral obstruction was maintained for 3 days than 1 day after urethral obstruction, although urinary beta2- microglobulin, Na, Cl, and osmotic pressure did not change during this period. Decompression of urethral obstruction 1 day after induction of urethral obstruction resulted in improvement in the severity glomerular hypoplasia compared with late decompression (P < 0.01). Our results suggest that the rabbit fetal model simulates fetal urethral obstruction in humans, and indicates that early surgical decompression may be effective in restoration of normal renal function.

Albuminuria↗

PubMed1998

Virus-associated haemophagocytic syndrome caused by rubella in an adult.

Haemophagocytic syndrome is a systemic clinicopathological entity characterized by systemic proliferation of benign haemophagocytic histiocytes, fever, cytopenia, abnormal liver function and, frequently, coagulopathy and hepatosplenomegaly. Its occurrence has been documented in association with viral, bacterial, fungal and parasitic infections, a wide spectrum of malignant neoplasms, autoimmune diseases and drugs. We report a case of rubella virus-associated haemophagocytic syndrome in a previously healthy 29-year-old woman. Blood tests showed cytopenia, especially severe thrombocytopenia, liver dysfunction, hyperferritinaemia and hypercytokinaemia. Bone marrow examination showed many mature histiocytes with active haemophagocytosis. A skin biopsy from the rash revealed perivascular lymphohistiocytic infiltrates with haemophagocytic histiocytes in the upper and mid-dermis. The patient was treated with antibiotics and immunoglobulin, and by supportive measures including platelet transfusion, and recovered completely.

Adult↗

PubMed1997

Isolation and characterization of the yeast mRNA capping enzyme beta subunit gene encoding RNA 5'-triphosphatase, which is essential for cell viability.

The yeast Saccharomyces cerevisiae mRNA capping enzyme is composed of two subunits of alpha (52 kDa, mRNA guanylyltransferase) and beta (80 kDa, RNA 5'-triphosphatase). We have isolated the alpha subunit gene (CEG1) by immunological screening. In this report, with the aid of partial amino acid sequences of purified yeast capping enzyme, we isolated the gene, designated CET1, encoding the S. cerevisiae capping enzyme beta subunit. Amino acid sequence analysis revealed that the gene encodes for 549 amino acids with a calculated M(r) of 61,800 which is unexpectedly smaller than the size estimated by SDS-PAGE. Gene disruption experiment showed that CET1 is essential for yeast cell growth. The purified recombinant CET1 gene product, Cet1, exhibited an RNA 5'-triphosphatase activity which specifically removed the gamma-phosphate from the triphosphate-terminated RNA substrate, but not from nucleoside triphosphates, confirming the identity of the gene. Interaction between the Cet1 and the Ceg1 was also studied by the West-Western procedure using recombinant Ceg1-[32P]GMP as probe.

3-Isopropylmalate Dehydrogenase↗

PubMed1997

Characterization of the response element and DNA binding properties of the nuclear orphan receptor germ cell nuclear factor/retinoid receptor-related testis-associated receptor.

Recently, we have reported the cloning of the germ cell-specific, nuclear orphan receptor germ cell nuclear factor (GCNF)/RTR. In this study, we characterize the RTR response elements by an electrophoretic mobility shift assay/polymerase chain reaction-based, DNA binding site selection strategy. RTR binds with the greatest affinity to response elements containing TCA(AG(G/T)TCA)2 (consensus RTR response element; conRTRE), to which it binds as a homodimer. RTR is also able to bind as a monomer to a single core motif TCAAG(G/T)TCA, albeit with a lower affinity. Mutation analysis supports the specific requirements of the 5'-flanking sequence and the core motif of the RTRE for optimal binding of RTR. An RTR-specific antiserum (RTR-Ab2) was raised that causes supershift of the RTR-conRTRE complex in EMSA. Based on the sequence of the conRTRE, we located a putative RTRE, referred to as P2-RE, in the 5' promoter-flanking region of the mouse protamine 2 gene, which is induced during the same stage of spermatogenesis as RTR. The ability of RTR-Ab2 to cause a supershift of an RTR-RTRE complex with nuclear extracts from different tissues correlated with the tissue- and development-specific expression of RTR. Transfection of RTR in CV-1 cells was unable to cause RTRE-dependent transactivation of a CAT reporter gene; however, an RTR-VP16 fusion protein could induce transactivation through several RTREs, including P2-RE.

Amino Acid Sequence↗

PubMed1997

Fetal redistribution of blood flow and amniotic fluid volume in growth-retarded fetuses.

Our purpose was to assess the relationship between the fetal redistribution of blood flow and the amount of amniotic fluid in appropriate-for gestational-age fetuses and growth-retarded fetuses. Blood flow velocity waveforms of the umbilical artery, descending aorta, middle cerebral artery, renal artery and uterine artery were recorded using pulsed Doppler ultrasonography in 100 appropriate-for gestational age fetuses and 39 growth-retarded fetuses. The pulsatility index (PI) values and the amount of amniotic fluid were compared between the two groups. The PI values of the umbilical artery and renal artery were significantly higher in appropriate for gestational-age-fetuses with oligohydraminos than in fetuses with an adequate amount of amniotic fluid. The PI values of the umbilical artery and renal artery were significantly higher and the PI of the middle cerebral artery was significantly lower in growth-retarded fetuses with oligohydramnios than in fetuses with an adequate amount of amniotic fluid. Furthermore, there was a significant negative correlation between the PI value of the renal artery and the vertical diameter of amniotic fluid, and between the PI value of the renal artery and the amniotic fluid index. The PI value of the renal artery was related to the amount of amniotic fluid in growth-retarded fetuses, and the same relationship was demonstrated in appropriate-for gestational age fetuses.

Amniotic Fluid↗

PubMed1997

The effects of oligohydramnios and cervical cord transection on lung growth in experimental pulmonary hypoplasia in rabbits.

OBJECTIVE: Our goal was to examine the effects of oligohydramnios and cervical cord transection on fetal lung development, pulmonary surfactant, and lung mechanics in rabbits. STUDY DESIGN: The amniotic fluid was shunted into the maternal abdominal cavity in a group of 12 fetal rabbits. In another group (n = 12) high cervical cord transection was performed at day 24 of gestation. Another 12 littermates not operated on served as the control group. Fetuses were delivered on day 30 of gestation by cesarean section and immediately put to death. The body weight and wet lung and liver weights were measured. To determine the extent of fetal lung growth, we measured the size of lung acini, the number of terminal airspaces, and the diameter of alveoli. We also measured the dynamic compliance of the lung, the concentration of phosphatidylcholine, and the lecithin/sphingomyelin ratio in lung lavage fluid at birth. RESULTS: Amniotic fluid shunting produced a significant reduction in amniotic fluid volume. Amniotic shunt and cervical cord transection significantly decreased wet lung weight and fetal lung/body weight ratio compared with the control. The concentration of phosphatidylcholine and the lecithin/sphingomyelin ratio in lung fluid lavage from fetuses with amniotic shunt were significantly higher than the values for control and cord transection fetuses. Histopathologic examination of the lungs showed significant reductions in the size of acini, the number of terminal airspaces, and the diameter of alveoli in shunted and cord transection groups compared with the control group. The dynamic compliance of transection fetuses was significantly reduced compared with control and shunted fetuses. CONCLUSION: Our results indicate that oligohydramnios or cervical cord transections cause pulmonary hypoplasia. However, oligohydramnios-induced pulmonary hypoplasia is associated with increased pulmonary surfactant compared with control and cord transection fetuses.

Animals↗