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Biomedical subjects

M Asakawa

Publications and source records attributed to M Asakawa.

At least 37 records · Page 2Linked to original sources
PubMed2000

Green fluorescent protein gene insertion of Sendai Virus infection in nude mice: possibility as an infection tracer.

The green fluorescent protein (GFP) marker from jellyfish Aequorea victoria is considered to have potential use in the study of host-pathogen relationships, by tracing infections in living cells, organs and animals. We compared the pathogenicity of Sendai virus with an inserted GFP gene (GFP-SeV) with that of its wild-type (Wt-SeV) to determine the usefulness of the recombinant virus in long-term infection of BALB/c nude (nu/nu) mice. The results indicated that the presence of GFP in infected cells could be analyzed easily and sensitively. GFP helped in identifying and in understanding the cellular sites of viral replication in vitro and in vivo. However, the GFP insertion into the Wt-SeV genome, led to decreased pathogenicity, altering the in vivo viral kinetics.

Animals↗
PubMed2000

[A morphological study of the kidney in the Japanese fetus with special reference to their external form, size and number of minor calix].

There are few anatomical reports on the fetal kidney than on adult human and animal kidneys. This paper reports the size, weight, ratio of renal hilus, number of minor calix and number of renal lobe in Japanese fetal kidneys. Fifty kidneys of 25 fetuses (3, 8, 8 and 6 bodies in 5, 6, 7 and 8 months, respectively) were used in this study. The length, width, thickness and weight of kidneys were measured, and the renal lobes were counted. The relative size of the hilus was then calculated by the ratio of the area of hilus to that of the medial surface of the kidneys. Latex rubber was injected in the ureters, taken out of the calico-pelvic system from the kidneys, and the number of minor calix in these specimens were counted. There were no significant sex difference and right and left difference in the kidney weight, ratio of renal hilus, number of minor calix and number of lobes. When these kidneys were observed at 5, 6, 7 and 8 months, the kidney weight and ratio of renal hilus were correlated with fetal age, but the numbers of lobe and minor calix were not. Comparing the results mentioned above with those of adult kidneys, it was shown that kidney weight and ratio of renal hilus area to the medial surface area of the kidneys gradually increased from fetus to adult. There was also a significant difference in the number of minor calix between adult and fetal kidneys. There was no correlation between the number of renal lobe and the number of minor calix.

Adult↗
PubMed1999

Accommodation of foreign genes into the Sendai virus genome: sizes of inserted genes and viral replication.

Sendai virus (SeV) is an enveloped virus with a negative sense genome RNA of about 15.3 kb. We previously established a system to recover an infectious virus entirely from SeV cDNA and illustrated the feasibility of using SeV as a novel expression vector. Here, we have attempted to insert a series of foreign genes into SeV of different lengths to learn how far SeV can accommodate extra genes and how the length of inserted genes affects viral replication in cells cultured in vitro and in the natural host, mice. We show that a gene up to 3.2 kb can be inserted and efficiently expressed and that the replication speed as well as the final virus titers in cell culture are proportionally reduced as the inserted gene length increases. In vivo, such a size-dependent effect was not very clear but a remarkably attenuated replication and pathogenicity were generally seen. Our data further confirmed reinforcement of foreign gene expression in vitro from the V(-) version of SeV in which the accessory V gene had been knocked out. Based on these results, we discuss the utility of SeV vector in terms of both efficiency and safety.

Animals↗
PubMed1999

The RAMP2/CRLR complex is a functional adrenomedullin receptor in human endothelial and vascular smooth muscle cells.

Adrenomedullin, a potently hypotensive peptide isolated from human pheochromocytoma, is known to elicit a rise in cAMP levels within mammalian endothelial and smooth muscle cells. Until now, however, little has been known about the adrenomedullin receptor. Recently, a group called receptor activity-modifying proteins that complex with the calcitonin receptor-like receptor, and thereby regulate its transport and ligand specificity, were identified. Here we show that mRNA for both the calcitonin receptor-like receptor and the receptor activity-modifying protein 2, but not the receptor activity-modifying protein 1 or receptor activity-modifying protein 3, are expressed in human endothelial and vascular smooth muscle cells. We also found that adrenomedullin increased cAMP levels in HeLa EBNA and 293 EBNA cells, expressing both the receptor activity-modifying protein 2 and calcitonin receptor-like receptor proteins. Thus, the receptor activity-modifying protein 2/calcitonin receptor-like receptor complex apparently serves as a functional adrenomedullin receptor in human endothelial and vascular smooth muscle cells.

Adrenomedullin↗
PubMed1999

First report of Thelazia sp. from a captive Oriental white stork (Ciconia boyciana) in Japan.

Nematodes of the genus Thelazia were recovered from the cornea and inferior conjunctival sac of an immature Oriental white stork (Ciconia boyciana). The bird hatched and reared at the Toyooka Oriental White Stork Breeding Center, Hyogo Prefecture, Japan, but died of chlamydiosis. There were neither gross nor histopathologic ophthalmic lesions. The eye worm from a bird is believed to be first reported in Japan. As regarding reintroduction plan for the Oriental white stork, control measures for prevent further infection with the eye worm will be needed.

Animals↗
PubMed1998

Measles virus attenuation associated with transcriptional impediment and a few amino acid changes in the polymerase and accessory proteins.

Measles virus (MV) isolated in B95a cells, a marmoset B-cell line, retains full pathogenicity for cynomolgus monkeys, while its derivative obtained by adaptation to the growth in Vero cells, a monkey kidney cell line, loses the pathogenic potential (F. Kobune, H. Sakata, and A. Sugiura, J. Virol. 64:700-705, 1990). Here, we show with a pair of strains, a fresh isolate (9301B) in B95a cells and its Vero cell-adapted form (9301V), that the in vivo attenuation parallels the decrease of replication and syncytium-inducing capabilities in the original B95a cells and that these in vitro phenotypes are attributable to impediment of transcription, which is already obvious at the level of primary transcription catalyzed by the virion-associated RNA polymerase. On the other hand, cell fusion assays detected no functional difference between the glycoproteins of the two viruses. Essentially the same transcriptional impediment with reduced syncytium induction following Vero cell adaptation was found with two other pairs of strains that had been similarly prepared. Nucleotide sequence comparison between the 9301B and 9301V viruses revealed that a few (at most five) amino acid changes, which sporadically took place in the polymerase (L and P proteins) and/or accessory V and C proteins, were responsible for the in vitro and in vivo attenuation through adaptation to growth in Vero cells.

Adaptation, Physiological↗
PubMed1998

[Left ventricular wall motion dynamics of asymmetric septal hypertrophy: assessment by intramyocardial pulsed Doppler echocardiography].

Apical pulsed Doppler tissue imaging can be used to assess the function of regional myocardium. We hypothesized that septal dysfunction might be clarified in the hypertrophic cardiomyopathy (asymmetric septal hypertrophy) by this method. Twenty-one patients with asymmetric septal hypertrophy (mean age 54.8 +/- 11 years) and age-matched 24 normal subjects (52.4 +/- 8 years) were studied. The E/A ratio measured by mitral inflow Doppler was not different between the groups (1.1 vs 1.2). E wave velocities of the septum were significantly decreased in the hypertrophy group compared to the control group (4.0 +/- 1.5 vs 8.1 +/- 2.2 cm/sec), and A wave velocities were increased in the hypertrophic septum, resulting in a significantly lower E/A ratio (0.5 +/- 0.3) compared to the E/A ratio (0.9 +/- 0.3) of the normal septum. Deceleration time of the E wave and isovolumic relaxation time were significantly prolonged in the thick septum compared to the normal septum (136 +/- 51 vs 107 +/- 28 msec, 91 +/- 36 vs 63 +/- 19 msec, respectively). In conclusion, asymmetric septal hypertrophy was characterized by diastolic dysfunction of the thickened septum. Intramyocardial pulsed Doppler echocardiography can detect regional myocardial dysfunction earlier than the mitral inflow Doppler method.

Cardiomyopathy, Hypertrophic↗
PubMed1998

[Ebstein's anomaly associated with left ventricular dysfunction: a case report].

A 66-year-old man presented with Ebstein's anomaly associated with left ventricular dysfunction. He had been followed since 40 years of age for cardiomegaly and arrhythmia, and experienced episodes of orthopnea at the age of 64. He was referred to our hospital in April 1997 because of lower extremity edema. Physical examination revealed dilated external jugular vein, tenderness of the right hypocondorium, and lower extremity edema. Electrocardiography confirmed atrial fibrillation. Transthoracic echocardiography revealed bilateral atrial and ventricular dilation, and paradoxical septal movement. The apical four-chamber view demonstrated 15 mm apical displacement of the septal leaflet. Color Doppler echocardiography revealed moderate tricuspid regurgitation. Transesophageal echocardiography revealed low echoic and hypoplastic tricuspid valve. Left ventriculography showed diffuse hypokinesis, and the ejection fraction was 49%. The coronary artery was normal. Atrial septal defect was not detected. Diffuse fibrosis, which may be found in the hearts of patients with Ebstein's anomaly at autopsy may have been responsible for the left ventricular depressed systolic function in this patient.

Aged↗
PubMed1997

Phosphorylation of the Sendai virus M protein is not essential for virus replication either in vitro or in vivo.

A large proportion of intracellular Sendai virus (SeV) M proteins is phosphorylated, but in mature virions the M protein is not phosphorylated or dephosphorylated. Phosphorylated M protein in cells is bound to the cytoskeletal components more firmly than unphosphorylated M protein. Thus it has been hypothesized that M protein phosphorylation plays an important role in the virus life cycle, especially in the step of maturation. Here, a transient expression-mutation experiment of the M gene demonstrated that a change of the Ser residue at the 70th position from the N-terminus to Ala (S70A) totally abolished M protein phosphorylation, strongly suggesting that this residue is phosphorylated. The mutated M gene was then placed in the corresponding region in the cDNA plasmid which generates a full-length antigenome SeV RNA, and a mutant SeV M-S70A was successfully recovered from the cDNA. This mutant virus was indeed defective in M protein phosphorylation but did not differ at all from the wild-type SeV recovered from the parental cDNA either in the replication kinetics and plaque morphology in cultured cells or in in vivo replication and pathogenicity for mice. We thus concluded that no phosphorylation of the M protein was required for SeV replication either in vitro or in vivo.

Animals↗
PubMed1997

Occurrence of paralytic shellfish poison in the starfish, Asterias amurensis in Kure Bay, Hiroshima Prefecture, Japan.

In May 1996, during surveillance on the toxicity of invertebrates such as bivalves inhabiting the coasts of Hiroshima Bay, the starfish Asterias amurensis collected in the estuary of the Nikoh River was found to contain toxins which showed strong paralytic action in mice; the maximum toxicity (as paralytic shellfish poison, PSP) was 8.0 MU/g for whole body and 28.7 MU/g for viscera throughout the monitoring period, March to July 1996. Attempts were made to identify the paralytic toxins in the starfish. They were extracted with 80% ethanol acidified with acetic acid, followed by defatting with dichloromethane. The aqueous layer obtained was treated with activated charcoal and then applied to a Sep-Pak C18 cartridge. The unbound toxic fraction was analyzed by high-performance liquid chromatography techniques. The starfish toxin was rather unexpectedly identified as PSP. It was comprised of high toxic components (gonyautoxin-1; GTX1, GTX2, GTX3, GTX4, decarbamoyl-GTX3; dcGTX3 and dcSTX) as the major components, which accounted for approximately 77 mole% of all components, along with protogonyautoxin-1, 2, 3 and 4 (PX1-4), which are N-sulfocarbamoyl derivatives. Of the high toxic components, GTX1 was present in the largest amounts. It was concluded that the toxin of starfish collected in the estuary of Nikoh River in May 1996 consisted of PSP, which supposedly came via the food chain from toxic bivalves living in the same area. To our knowledge, this is the first report of the occurrence of PSP in starfish.

Animals↗
PubMed1997

cis-Acting inhibitory elements within the pol-env region of human T-cell leukemia virus type 1 possibly involved in viral persistence.

Human T-cell leukemia virus type 1 (HTLV-1) remains latent throughout the life of the carrier, with cells containing the provirus and viral gene expression efficiently down-regulated. On a molecular level, exactly how viruses are down-regulated in vivo remains unresolved. We described here the possibility that down-regulation results from the presence of inhibitory elements within the gag-env region of the provirus in fresh peripheral blood mononuclear cells from carriers. In vitro experiments then revealed that potent cis-acting inhibitory elements (CIEs) are indeed contained in two discrete fragments from the pol region and weaker ones in the env region. The effect of CIEs is relieved by the HTLV-1 posttranscriptional regulator Rex through binding to the Rex-responsive element (RxRE), suggesting that Rex might interfere with pre-mRNA degradation and/or activate the export of mRNA molecules harboring both of the inhibitory elements and RxRE on the same RNA molecule. Thus, we propose the hypothesis that such functions of CIEs may be involved in HTLV-1 persistence.

Cell Line↗
PubMed1997

First record of Eustrongylides tubifex (Dioctophymatidae) from little grebe, Tachybaptus ruficollis in Japan.

A nematode species belonging to the genus Eustrongylides (Dioctophymatidae) was obtained from the proventriculus of Tachybaptus ruficollis, found in December, 1995 in Kobe City, Hyogo Prefecture, Japan. Observation under a scanning electron microscope to investigate anterior extremity with the labial papillae in two circles showed that this species is identical to E. tubifex. This is the first record of this species in Japan.

Animals↗
PubMed1997

[An immunohistochemical study on glycosaminoglycans distribution in the trabeculum of congenital aniridia].

Distribution of glycosaminoglycans in the trabecular tissue was immunohistochemically investigated in 7 congenital aniridia eyes of 5 patients aged 0 to 43 days. Paraffin sections of each specimen were immunohistochemically stained with antibodies to chondoroitin (clone 1-B-5), chondoroitin-1-sulfate (2-B-6), chondoroitin-6-sulfate (3-B-3), dermatan sulfate (6-B-6), and keratan sulfate (5-D-4). The trabecular meshwork and Schlemm's canal in all eyes were absent or not well differentiated. The cornea, trabecular tissue, and iris stroma were negative for chondoroitin immunostaining but positive for chondoroitin-4-sulfate, chondoroitin-6-sulfate, dermatan sulfate and keratan sulfate immunostaining. In the normal anterior segment tissue keratan sulfate is present in the cornea, trabecular tissue, and iris at the fetal stage, and disappears from the iris at the neonatal stage. These findings suggest that the persistence and/or abnormal distribution of keratan sulfate in the anterior segment may play a role in the pathogenesis of congenital aniridia.

Aniridia↗
PubMed1995

Dinoflagellate Alexandrium tamarense as the source of paralytic shellfish poison (PSP) contained in bivalves from Hiroshima Bay, Hiroshima Prefecture, Japan.

In April 1993, a phytoplankton dinoflagellate was isolated from Hiroshima Bay, Hiroshima Prefecture, Japan, and unambiguously identified as Alexandrium tamarense on the basis of the morphological characteristics. The dinoflagellates, cultures in modified SW-2 medium at 15 degrees C for 15 days, showed a specific toxicity of 30.7 x 10-6 MU/cell. HPLC analysis demonstrated that the toxin was composed mainly of gonyautoxin-4 (GTX4) and protogonyautoxin-2 (PX2 or GTX8)(27.6 and 37.0 mole%, respectively). Total toxin concentration of this strain was 39.5 fmole/cell. Short-necked clams, mussels, and oysters contaminated by the dinoflagellate showed a more complicated composition, with GTX1 as the major component (61.8 mole% for short-necked clams, 60.5 mole% for mussels, 42.5 mole% for oysters), and PX2 was only present in trace amounts.

Animals↗
PubMed1995

[A case of malignant lymphoma arising from chronic pyothorax; usefulness of magnetic resonance imaging (MRI) for diagnosis].

A 79-year-old man had a history of pneumothorax induced to treat pulmonary tuberculosis when he was 40 years old, and of chronic pyothorax when he was 60 years old. He was admitted to our hospital because of chest pain and swelling of the right lateral chest wall. Soft tissues of the chest wall and rib had been destroyed, and a fistula had formed at the skin. The diagnosis was confirmed by examination of a biopsy specimen from the chest wall tumor. The tumor was successfully treated by radiation therapy. 67Ga scintigraphy, CT, and MRI were useful for diagnosis and follow up. Pyothorax and the tumor could be distinguished only with MRI.

Aged↗
PubMed1994

Characterization of a deaminated neuraminic acid-containing glycoprotein from the skin mucus of the loach, Misgurnus anguillicaudatus.

Using NMR spectroscopy and mass spectrometry, the major sialic acid of the skin mucus of the loach, Misgurnus anguillicaudatus was found to be 3-deoxy-D-glycero-D-galacto-2-nonulosonic acid (KDN). We have subsequently devised a method to isolate a KDN-containing glycoprotein preparation from loach skin mucus. The method involves the sonication of the skin mucus with 0.05 M Tris-HCl, pH 8.0, to solubilize the glycoprotein, followed by DE52-cellulose chromatography of the extract, Nuclease P1 treatment, and Sepharose CL-4B gel filtration. The purified glycoprotein preparation was found to contain 38.5% KDN, 0.4% NeuAc, 24.6% GalNAc, 3.3% Gal, and 28.2% amino acids (w/w). The amino acid composition of this glycoprotein preparation revealed that it is unusually rich in Thr, and 6 amino acids, Thr, Ser, Glu (or Gln), Pro, Gly and Ala, account for 83% of the total amino acids. This glycoprotein is extremely poor in Cys, Met, Tyr, Phe, Arg, and Trp. Treatment of this glycoprotein with alkali resulted in the destruction of 83% of Thr suggesting that most of the sugar chains in this glycoprotein are linked through Thr. Alkaline borohydride treatment of 100 mg of the glycoprotein preparation, followed by Sephadex G-25 (superfine) gel filtration, yielded two major oligosaccharide alditols, I (15.4 mg) and II (15.6 mg). Using liquid secondary ion mass spectrometry and methylation analysis, I was identified to be KDN alpha 2-->6GalNAc-ol and II, KDN alpha 2-->6(KDN alpha 2-->3)-GalNAc-o1. KDN alpha 2-->6GalNAc is structurally similar to NeuAc alpha 2-->GalNAc found in ovine submaxillary glycoprotein while II represents a novel structure which contains two sialic acids linked to a GalNAc through both alpha 2-->3 and alpha 2-->6 linkages. This structure has never been found in mucin type glycoproteins including mammalian epithelial mucin glycoproteins. This is the first report of the presence of a mucin type glycoprotein which contains KDN instead of NeuAc or NeuGc in fish skin mucus.

Amino Acids↗