Search PubMed⌕ Search

Biomedical subjects

T Haga

Publications and source records attributed to T Haga.

At least 109 records · Page 6Linked to original sources

Molecular analysis of virus-producing and non-producing clones derived from a defective SSPE virus Yamagata-1 strain.

Two virus clones were isolated from a defective SSPE virus, the Yamagata-1 strain, and designated as the YA and YF clones. The YA clone-infected cells produced neither cell-free virus nor cell-associated virus, whereas the YF clone-infected cells produced both cell-associated and cell-free virus. No difference of epitopes on structural proteins was observed between these two clones. Both clones had hemadsorption activity. Quantitation of structural protein by Western dot blots showed relatively a lower amount of M protein in the YA-infected cells than that in the YF-infected cells. The ratio, P plus M dicistronic/M monocistronic mRNA, in the YA-infected cells was about twice that in the YF-infected cells. Sequence analysis of cDNA corresponding to P plus M dicistronic mRNA revealed that the deduced M protein of the YF virus was smaller than that of the YA virus by five amino acids from the carboxy terminal. These results suggest that abundant production of P plus M dicistronic mRNA is responsible for the reduced amount of M protein in the non-productive YA clone.

Amino Acid Sequence↗

[Recent technical advances in portable oxygen delivery systems].

According to a Japanese national survey (June 30, 1990), the number of patients receiving home oxygen therapy (HOT) has been greater than 18,000 since March 1985, when HOT was first covered by health insurance. The oxygen concentrator, especially the molecular sieve type, is the most common method of delivery (more than 90%). In April 1988, the portable oxygen cylinder was acknowledged by health insurance, and the liquid oxygen supply system in April 1990. Three types of portable oxygen delivery systems are available; oxygen cyclinder, liquid oxygen system, and oxygen concentrator (membrane type), of which the oxygen cylinder is most commonly used. In our hospital, portable oxygen supply systems were used in 80% of 168 HOT cases in 1990, and the use of 400 L aluminum oxygen cylinders at a flow rate of 1-2 L/min has been most popular. There is an strong desire from patients for lighter portable oxygen supply system of longer duration. In 19 patients with chronic respiratory failure, we evaluated a newly designed demand oxygen delivery system (DODS), which weighs 2.4 kg including the DOD device (TER-20 Teijin), 1.1 L oxygen cylinder made of ultressor, nasal cannula, and carrier. Arterial blood gases at rest (room air) were PaO2 61.9 +/- 6.3 torr, PaCO2 63.8 +/- 9.4 torr and pH 7.40 +/- 0.04. A crossover trial was performed under three conditions; breathing room air with no weight, and pulse oxygen flow and continuous oxygen flow each carrying 2.4 kg of weight. Both 6 minute walking (E1) and walking on a slow speed treadmill (E2) were studied.(ABSTRACT TRUNCATED AT 250 WORDS)

Female↗

MR imaging of chronic alcoholism.

We evaluated the brain lesions of patients with chronic alcoholism (n = 34) in comparison with age- and sex-matched controls (n = 40) by MR imaging. T1-weighted sagittal and axial images and T2-weighted axial images were obtained with a 0.5 T superconducting MR unit. Various brain measurements were then performed, and the presence of regions of abnormal signal intensity was also compared between the two groups. The brain measurements revealed significant cerebral atrophy (characterized by lateral and 3rd ventricular dilatation, and widening of the interhemispheric fissure) as well as significant cerebellar atrophy (represented by 4th ventricular dilatation) in the alcoholic group. These changes were more prominent in patients in their fifties and sixties than in those aged in the thirties and forties. Focal hypointense lesions were observed in 20.6% of the alcoholics and in 5% of the controls (p less than 0.01), while focal hyperintense lesions were observed in 61.8% of the alcoholics and in 20% of the controls (p less than 0.001). The severity of these MR findings correlated well with the age of the patients. These observations suggest that alcohol is an important promoter of brain aging.

Adult↗

Identification of two novel GTP-binding protein alpha-subunits that lack apparent ADP-ribosylation sites for pertussis toxin.

Molecular cloning of cDNAs encoding alpha-subunits of guanine nucleotide-binding regulatory proteins (G-proteins) has revealed the existence of nine species of alpha-subunits. We have identified two additional G-protein alpha-subunits, which we refer to as GL1 alpha and GL2 alpha, by isolating bovine liver cDNA clones that cross-hybridized at reduced stringency with bovine Gi1 alpha-subunit cDNA. The deduced amino acid sequences of GL1 alpha and GL2 alpha share 83% identity with each other and show 45-55% identity with those of other known G-protein alpha-subunits. Both GL1 alpha and GL2 alpha lack a consensus site for ADP-ribosylation by pertussis toxin. Messenger RNA corresponding to GL2 alpha was detected in all tissues examined, but GL1 alpha mRNA was detected only in liver, lung, and kidney. Antiserum prepared against a synthetic pentadecapeptide corresponding to the deduced carboxyl terminus of GL2 alpha specifically reacted with a 40-kDa protein in mouse liver, brain, lung, heart, kidney, and spleen. The amount of the 40-kDa protein was highest in brain and lung. We suggest that GL1 alpha and GL2 alpha are new members of a subfamily of pertussis toxin-insensitive G-proteins.

Adenosine Diphosphate Ribose↗

Synthesis and antitumor activities of novel benzoylphenylurea derivatives.

Seventy novel benzoylphenylurea compounds were synthesized and their antitumor activities were examined in vivo against P388 leukemia. N-(2-Nitrobenzoyl)-N'-[4-(2-pyrimidinyloxy)phenyl]ureas showed the highest antitumor activities when dosed intraperitoneally or orally. Their structure-activity relationships were examined with particular focus on the position and the variety of substituent on each aryl ring.

Animals↗

[Mycobacterium examination system at the National Jewish Center in the States].

Impressions on visit to Mycobacteriology Laboratory of National Jewish Center for Immunology and Respiratory Medicine are reported. The BACTEC and Gen-Probe Method are introduced to the laboratory system on mycobacterial examinations such as detection, identification, drug susceptibility and minimal inhibitory concentration test.

Bacteriological Techniques↗

[What has tuberculosis medical care contributed to modern medicine in Japan].

About 40 years ago, we had a huge number of tuberculosis patients which were about three to five millions. And a great deal of effort has been given in order to conquer tuberculosis. The philosophy and the technology used in there efforts are thought to have made considerable contributions to the development of modern medicine in other fields in Japan. The controlled clinical trial used to test new antituberculous agents has been a basis for a modern judgment on the effectiveness of any new drugs and it has brought good clinical practices. Scientific procedures for diagnosis, bronchoscopy and pulmonary function tests started in the field of tuberculosis to have more detailed diagnosis included differential one, preoperative evaluation, and evaluation for respiratory failure. Philosophy and methods of modern rehabilitation for the patients started from the care for the patient with pulmonary tuberculosis. Occupational therapy has been used for confirmation of curability of tuberculosis and restoration of the physical strength. Physiotherapy also started from exercises preventing deformities of the thorax after thoracoplasty. Terminal care for patients with cancer and other incurable diseases is now most new medical way, but in the field of tuberculosis before chemotherapeutic era, these were good experiences for this kind of care. A pain control using morphine or cocaine for spinal caries and laryngeal tuberculosis, and also telling the truth to the patients were the good examples. Holistic medicine, which is also a modern topic, has been considered in tuberculosis medical care, because the disease has so much psycho-social problems.(ABSTRACT TRUNCATED AT 250 WORDS)

Clinical Medicine↗

Interaction of deglycosylated muscarinic receptors with ligands and G proteins.

Endoglycosidase F was used to investigate the role of the carbohydrate moiety of muscarinic acetylcholine receptors in antagonist and agonist binding, and the interaction with G proteins. The receptors were purified from porcine cerebrum, treated with endoglycosidase F and then covalently labeled with [3H]propylbenzilylcholine mustard [( 3H]PrBCM). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the [3H]PrBCM-labeled receptors showed that the endoglycosidase F treatment caused a decrease in apparent Mr from 70 to 51 kDa, the Mr predicted for the peptide portions of M1, M2 and M4 subtypes. Endoglycosidase F-treated receptors had essentially the same affinities for both agonists and antagonists as those of control receptors. In addition, treated receptors that had been reconstituted in lipid vesicles with G proteins showed guanine nucleotide-sensitive high affinity for agonists. These results suggest that the carbohydrate moiety of muscarinic acetylcholine receptors is not involved in their interaction with muscarinic ligands and G proteins.

Animals↗

Dual regulation by G proteins of agonist-dependent phosphorylation of muscarinic acetylcholine receptors.

Muscarinic acetylcholine receptors purified from porcine atrium were phosphorylated, depending on the presence of agonists, by a protein kinase partially purified from porcine brain, which had similar properties to the beta-adrenergic receptor kinase. GTP-binding regulatory proteins (Go) had dual effects on the phosphorylation of muscarinic receptors, i.e. stimulation at lower concentrations and inhibition at higher concentrations. The stimulatory effect was reproduced with the beta gamma subunit of Go and the inhibitory effect with the combination of the alpha and beta gamma subunits.

Adenosine Triphosphate↗

Carbachol-induced potentiation and inhibition of acid secretion by guinea pig gastric gland.

The effects of muscarinic ligands on acid secretion were examined by estimating the accumulation of [14C]aminopyrine in gastric glands isolated from guinea pigs. The accumulation of [14C]aminopyrine in the presence of 0.1 mM histamine was potentiated by 1 microM carbachol but suppressed by 1 mM. These two effects of carbachol were abolished by atropine, pirenzepine and AF-DX 116. Assuming that the binding of carbachol to one site (Site 1) increases [14C]aminopyrine accumulation but its binding to the other site (Site 2) reduces [14C]aminopyrine accumulation, we analysed the dose-response curves for the carbachol effects in the absence and presence of different concentrations of atropine, pirenzepine and AF-DX 116. The dissociation constants determined for these ligands at Sites 1 and 2 were as follows: carbachol, 0.28 and 7.1 microM; atropine, 0.28 and 0.54 nM; pirenzepine, 45 and 560 nM; and AF-DX 116, 380 and 4400 nM, respectively. The binding of [3H]N-methylscopolamine to the gastric glands indicated the presence of two populations of binding sites with different affinities for the above ligands, other than atropine. The apparent dissociation constants, which were estimated by analysing the displacement curves for [3H]N-methylscopolamine binding, were as follows: carbachol, 0.18 microM (10%) and 31 microM (90%); atropine, 1.24 nM; pirenzepine, 15 nM (16%) and 220 nM (84%); and AF-DX 116, 370 nM (10%) and 2970 nM (90%). These results suggest that there are two kinds of muscarinic acetylcholine receptors in the guinea pig gastric gland, one potentiating and the other inhibiting the acid secretion induced by histamine.

Aminopyrine↗

Molecular analysis of structural protein genes of the Yamagata-1 strain of defective subacute sclerosing panencephalitis virus. II. Nucleotide sequence of a cDNA corresponding to the P plus M dicistronic mRNA.

The nucleotide sequence of a cloned cDNA corresponding to the P + M dicistronic mRNA of a subacute sclerosing panencephalitis (SSPE) virus was determined and compared with data of measles virus (MV). The dicistronic mRNA of the SSPE virus consisted of the 3' proximal 626 nucleotides of P mRNA, intercistronic trinucleotides, a full length of M mRNA, and 75 poly A nucleotides. The part encoding the P protein had a high homology to MV, except at the noncoding region. The terminating consensus sequence of the P gene and the intercistronic trinucleotides of the SSPE virus were CTAC(A)6 and CCT; in MV they are TTAT(A)6 and CTT, respectively. In the M gene, the starting consensus sequence was exactly the same as MV, but at the 5' proximal end, one third of this gene was different: The first ATG codon of the MV M gene signaling opening of the reading frame was changed to ACG in the SSPE virus and one long open reading frame started from the third ATG codon. The stop codon (TAG) of the MV M gene was also changed to CAG in the SSPE virus. Thus, the deduced SSPE-virus M protein lacked 50 amino acids at the amino terminal and had 15 extra amino acids at the carboxyl end when compared with the MV M protein.

Amino Acid Sequence↗

Molecular analysis of structural protein genes of the Yamagata-1 strain of defective subacute sclerosing panencephalitis virus. III. Nucleotide sequence of the hemagglutinin gene.

The full-length cDNA corresponding to the mRNA for the hemagglutinin (H) protein of the Yamagata-1 strain of the subacute sclerosing panencephalitis (SSPE) virus was cloned and the nucleotide sequence was determined. The mRNA corresponding to the H protein was composed of 1952 nucleotides and contained a single large open reading frame, which encoded 620 amino acids with a predicted molecular weight of 69,723. This cDNA clone expressed the H protein in Cos 7 cells, and the transfected cells showed hemadsorption. The nucleotide and amino-acid sequence homology with the Edmonston strain of MV were 98.0% and 96.6%, respectively. The deduced amino acid sequence had a single hydrophobic domain near the N-terminus that was long enough to serve as an anchor in the membrane. Five potential glycosylation sites were found on the H protein at identical positions as in the H protein of MV. Cysteine and proline were located at almost identical positions as those of the H protein of MV. In addition, monoclonal antibody study revealed that three epitopes, including the domains that were involved in the biological activities of the H protein of MV, were conserved on the Yamagata-1 strain. These results suggested that the H protein of the Yamagata-1 strain of defective SSPE virus is structurally and functionally similar to that of the Edmonston strain of MV.

Amino Acid Sequence↗

Molecular analysis of structural protein genes of the Yamagata-1 strain of defective subacute sclerosing panencephalitis virus. IV. Nucleotide sequence of the fusion gene.

The full-length cDNA corresponding to the mRNA of the fusion (F) protein of the Yamagata-1 strain of subacute sclerosing panencephalitis (SSPE) virus was cloned, and its complete nucleotide sequence was determined. The F gene was composed of 2369 nucleotides and contained a single large coding region, which is located between two noncoding regions. The 5'-terminal noncoding region consisted of 584 nucleotides comprising 44.9% cytosine, and had several inverted repetitious sequences. The 3'-terminal noncoding region had a relatively low homology of 91.7% with the MV. The coding region was expanded for nucleotides 585-2189, which encoded 534 amino acids with a molecular weight of 57,963. The homology of the amino acid sequence of the F protein between the MV and SSPE virus was 96.27%, and the positions of cysteine and proline were almost identical in the two viruses. The functional domains of SSPE-virus F protein closely resembled those of MV F protein, including the cleavage site, a signal sequence, the fusion-related stretch, the transmembrane region, and four potential glycosylation sites. Four antigenic epitopes on the MV F protein were also conserved on the SSPE-virus F protein. However, deletion of one nucleotide (position 2155) of the SSPE virus was found when compared with the MV, and shifted the coding frame, causing the substitutions of 27 C-terminal amino acids of the MV F protein with 11 different residues. The variations of the C-terminal region of the F protein were observed with two other SSPE viruses, suggesting that this may be a common property of SSPE virus that differs from MV.

Amino Acid Sequence↗

Interaction of atrial muscarinic receptors with three kinds of GTP-binding proteins.

Purified porcine atrial muscarinic acetylcholine receptors were reconstituted into lipid vesicles with three different G proteins (Gi, Go and Gn)1 purified from porcine cerebrum. All the G proteins interacted with the receptor as evidenced by GTP-sensitive high affinity binding with acetylcholine, and stimulation by acetylcholine of GTP gamma S binding and GTPase activities. The curves of displacement by acetylcholine of [3H]QNB binding were explained by assuming two sites with the same affinity for [3H]QNB but different affinities for acetylcholine. The proportion of the high affinity site increased from 3 to 7% up to 82 to 83% of total binding sites with increasing G protein concentration, and essentially the same results were obtained with the three G proteins. The GTPase activities of Gi, Go and Gn in the reconstituted vesicles were 2.7-, 1.7- and 1.6-times higher, respectively, in the presence of 1 mM acetylcholine than those in the presence of 10 microM atropine. An obvious enhancement by acetylcholine of the GTP gamma S binding was observed in the presence of 10 to 100 microM GDP, while the enhancement was minimal, if at all, in the absence of GDP. When the molar ratios of reconstituted Gi, Go and Gn to muscarinic receptors were 54, 84 and 107, respectively, the acetylcholine-induced increase in the [35S]GTP gamma S binding was as much as 12, 35 and 27 mol with Gi, Go and Gn, respectively, per mole of the receptor molecule, indicating that the muscarinic receptors interact with G proteins catalytically.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗

Diversity of matrix protein in subacute sclerosing panencephalitis and measles virus-infected cells.

Expression of the viral matrix (M) proteins in Vero cells infected with 18 strains of subacute sclerosing panencephalitis (SSPE) virus and measles virus was examined by immunocytochemistry and Western blot analysis using an anti-M monospecific serum and two sera against the M protein specific synthetic peptides. By immunocytochemistry using the anti-M monospecific serum, M protein was detected in all of the virus-infected cells regardless of cell-free virus production. M proteins of the seven non-productive strains were found to vary significantly in their epitope, in their reactivity to different assay systems, and in their molecular weight, whereas M proteins of the other 11 productive strains were detected consistently. These results suggest diversification of M protein of the non-productive strains.

Amino Acid Sequence↗