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Biomedical subjects

S Watabe

Publications and source records attributed to S Watabe.

At least 109 records · Page 6Linked to original sources

Occurrence of chondroitin sulfate E in glycosaminoglycan isolated from the body wall of sea cucumber Stichopus japonicus.

Glycosaminoglycan was isolated from the body wall of sea cucumber Stichopus japonicus by a method consisting of enzymatic digestion, gel filtration, and ion-exchange chromatography. One gram of sea cucumber glycosaminoglycan was composed of 2.50 mmol of sulfate, 0.47 mmol of N-acetylgalactosamine (GalNAc), 0.53 mmol of glucuronic acid (GlcA), 1.73 mmol of fucose, and a small amount of peptide. When mildly hydrolyzed with 0.1 N H2SO4, this glycosaminoglycan released two products, one consisting of fucose plus sulfate and the other of fucose only. Partially hydrolyzed glycosaminoglycan thus obtained was composed of sulfate, GalNAc, GlcA, and fucose at a molar ratio of 3:2:2:1. Partially hydrolyzed glycosaminoglycan was easily digested with chondroitinase AC II. In ion-exchange chromatography, the digest exhibited four sharp peaks whose retention times agreed with those of unsaturated 0-(delta Di-0S), mono-(delta Di-4S and delta Di-6S), and di-(delta Di-SE) sulfated disaccharide, respectively. The disaccharide unit of sea cucumber glycosaminoglycan was composed of 22.4% chondroitin sulfate E, 11.2% chondroitin, 10.4% chondroitin 4-sulfate, and 56.0% chondroitin 6-sulfate.

Animals↗

Clear and colorless fluid observed during percutaneous transhepatic gallbladder drainage.

Twelve patients with discharge of clear colorless fluid during percutaneous transhepatic gallbladder drainage (PTGBD) were encountered during a period of 4.5 years. On the average, the fluid appeared on the 12th day after PTGBD and continued to flow until the completion of observation. The volume was usually less than 60ml per day. It flowed in an alternating pattern with normal yellow color bile each day. The fluid was observed in patients in whom satisfactory patency of the biliary system was confirmed by cholangiography and/or cholangio-fiberscopy. Biochemical examinations of the fluid revealed lower biliary lipids but a similar electrolyte composition compared to bile of normal color. It was alkaline. These observations indicate that the clear colorless fluid is different from what is known as "white bile", which is produced when there is an obstruction of the biliary tree. The fluid may be secreted from the gallbladder epithelium or the bile duct epithelium. The significance was not clarified in this study. Further investigation is necessary to elucidate the clinical implications of the clear and colorless fluid secretion.

Bile↗

Alkali light chains are involved in stabilization of myosin head.

1. Fast skeletal myosin subfragment 1 (S1) was separated into two isozymes, S1(A1) and S1(A2), based on the associated alkali light chain, and their thermostabilities were compared. 2. Inactivation rate constants of Ca2(+)-ATPase (at 30 and 35 degrees C) were higher and heat-induced turbidity increase at 340 nm (at 40 degrees C) was faster with S1(A1) than with S1(A2), indicating a higher stability of S1(A2). 3. When S1 isozymes were incubated in the presence of excess alkali light chain, turbidity increase was markedly reduced, depending on the amount of light chain added. 4. Results obtained strongly suggest that alkali light chains are involved in the maintenance of myosin head structure.

Animals↗

Application of the Karhunen-Loeve expansion to evaluate regional cardiac excitation in body surface potential maps.

The authors investigated the usefulness of the Karhunen-Loeve technique applied to body surface maps to study regional cardiac excitation. Eigenvectors were derived from the body surface potential maps of 120 healthy adults using the Karhunen-Loeve expansion theory. Then, in the maps of various types of ventricular hypertrophy, each eigenvector coefficient was calculated for a statistical comparison. The first eigenvector coefficient in early QRS and the second in mid QRS were larger in patients with asymmetrical septal hypertrophy and in patients with left ventricular hypertrophy, respectively. The third was larger in patients with right ventricular hypertrophy. In the maps of patients with previous anteroseptal myocardial infarction, the second eigenvector coefficient decreased with asynergy of the anterior to apical wall, and the first decreased with the asynergy of the interventricular septum. They conclude that some eigenvector components and coefficients at particular times in the QRS are sensitive to changes in regional cardiac excitation and that they may facilitate the detection of local excitation changes such as occur in hypertrophy or infarction.

Adult↗

Mapping of fish myosin light chains by two-dimensional gel electrophoresis.

1. Myosins were prepared from the ordinary muscle of 16 fish species as well as from rabbit fast muscle, and light chain subunits [alkali light chains A1, A2 and DTNB (5,5'-dithio-bis-2-nitrobenzoate) light chain] were separated on two-dimensional gel electrophoresis in combination with isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. 2. A1 light chains showed mol. wts ranging from 21,000 to 22,900 and isoelectric points ranging from 4.51 to 4.62. DTNB light chains were spotted in a narrow area, with a mol. wt range of 16,800-17,600 and an isoelectric point range of 4.48-4.55. On the other hand, A2 light chains were most species-specific, with a mol. wt range of 14,000-19,500 and an isoelectric point range of 4.31-4.46. 3. It was suggested that the lower species-specificity in A1 as opposed to A2 is accounted for by the addition of an N-terminal peptide ("difference peptide") in the former. The properties and possible role of this peptide are discussed.

Animals↗

Possible presence of the difference peptide in alkali light chain 1 of fish fast skeletal myosin.

1. Presence of N-terminal peptide ("difference peptide") in alkali light chain 1 (A1) of fish fast skeletal myosin was examined by comparing two kinds of light chain-based myosin subfragment 1 (S1) isozymes from the yellowtail Seriola quinqueradiata. 2. On tryptic digestion, A1 was cleaved to a smaller fragment (mol. wt decrement by 2000) along with the cleavage of S1 heavy chain, while A2 was resistant to trypsin. Two-dimensional gel electrophoresis showed that A1 released a basic peptide by tryptic digestion. 3. Both S1 isozymes showed clear kinetic differences in actin-activated Mg-ATPase activity, suggesting a higher affinity of A1 for actin. Affinity of A2 for heavy chain was also estimated to be about 2-fold higher than that of A1, as judged by the model experiments in which rabbit S1 isozymes were hybridized with heterologous alkali light chains.

Actins↗

Paramyosin and the catch mechanism.

1. Catch is a mechanism found in many molluscan smooth muscles in which tension is maintained at relatively low energy cost. 2. Paramyosin forms the core of thick filaments. In catch muscle paramyosin concentrations are high and the thick filaments are relatively long. 3. The mechanism of catch is not understood, but the consensus is that tension during catch is borne by slowly-cycling cross-bridge attachments to actin. 4. Stimulation by acetylcholine increases intracellular Ca2+ and initiates a contraction characterized by a relatively rapid cross-bridge cycling. Reduction of Ca2+ can lead to relaxation or catch. Relaxation occurs only when a second neurotransmitter, serotonin, is present. 5. The catch state is released by serotonin, via activation of adenylate cyclase, increased levels of cAMP and phosphorylation of one or more contractile proteins, possibly paramyosin. Other targets for phosphorylation are discussed. 6. The contractile cycle of catch muscles, therefore, is controlled by both Ca2+ and cAMP.

Acetylcholine↗

Auditory sound agnosia without aphasia following a right temporal lobe lesion.

A 55-year-old right-handed man showed inability to recognize the meaning of non-verbal sounds without impairment of language comprehension after a cerebrovascular accident. His auditory acuity was intact and no other sign of agnosia, apraxia or aphasia was detectable. His errors on a test of sound recognition were acoustic rather than semantic. Brain CT scan showed a small lesion in the posterior part of the right temporal lobe. This case suggests that auditory sound agnosia without language disorder can ensure a lesion confined to the right hemisphere, and that the deficit is discriminative rather than associative in nature.

Agnosia↗

Effect of nontransmural necrosis on epicardial potential fields. Correlation with fiber direction.

The effect of nontransmural necrosis on epicardial potential distributions was studied in 13 dogs. In previous studies, left ventricular epicardial pacing generated epicardial potential maps at QRS onset with a negative central area and two positive areas that faced the portions of the wavefront propagating along fibers. Subsequently, the positive areas expanded in a counterclockwise direction by 90 degrees to 120 degrees. In those studies, the rotatory expansion of the positive areas was tentatively attributed to the spread of excitation through deep myocardial layers, where fiber direction rotated counterclockwise from epicardium to endocardium. To test this hypothesis, we tried to interrupt the counterclockwise expansion of the positive area by creating localized, nontransmural necrosis at various depths in the left ventricular wall by injection of formalin or application of laser energy. Epicardial potential maps were obtained from a grid of 12 x 15 electrodes on a 44 x 56-mm area. Epicardial pacing from selected sites generated epicardial maps in which some positive areas were missing compared with controls. The direction of the straight line joining the pacing site to the site of missing positivity correlated well with the average fiber direction in the necrotic mass (r = 0.82, p less than 0.01). Angle between epicardial fiber direction and the straight line described above correlated well with the average depth of the necrosis, expressed as percent of the wall thickness (r = 0.95, p less than 0.01). These data support the hypothesis that the counterclockwise expansion of the epicardial positivity occurring after epicardial pacing results from excitation spreading along deep fibers.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Glycosaminoglycan involved in the cation-induced change of body wall structure of sea cucumber Stichopus japonicus.

The body wall of sea cucumber Stichopus japonicus was treated with various concentrations of several cations, and examined for changes in toughness, taking punch force as parameter. Toughness of the body wall tended to decrease with increasing concentration of each cation, but in different modes depending mainly upon the valency of cation: e.g., the body wall completely lost toughness in 0.3 M Na+ or 0.4 M K+, whereas it retained more than half the initial toughness even in 0.4 M Ca2+ or Mg2+. Glycosaminoglycan (GAG) from the body wall was dissolved in water, and examined for viscosity changes as caused by those cations. Specific viscosity (eta sp) decreased from 0.71 (without cation) to 0.47-0.57 in the presence of 0.1 M monovalent and divalent cations. At 0.4 M, monovalent cations reduced eta sp to 0.38-0.46, but divalent cations increased eta sp to 0.56-0.63. Electron microscopy demonstrated that GAG matrix was clearly observed in the absence of cation, but disappeared in 0.4 M NaCl, forming wide free spaces in the body wall. These results all suggested that GAG is closely involved in the change of toughness of sea cucumber body wall.

Animals↗

[A recurrent gastric cancer improved by radiation therapy].

In March, 1988, a 65 year-old male underwent a total gastrectomy for a gastric cancer (Borrmann 3: a poorly differentiated adenocarcinoma; IIc: signet ring cell carcinoma). In October that year, however, an anastomotic recurrence of this tumor was found. Thus, from November, 1988 to January, 1989, to treat this recurrence, the patient received a course of radiation therapy frontally, towards the upper abdomen, with a total dose of 46.5 Gy (linac X-ray). His symptoms improved, and a CT, an upper gastrointestinal barium study, and an endoscopic examination indicated that the tumor had disappeared. No further recurrence was found until January, 1990, when the patient died of a metastasis that had spread throughout his body.

Adenocarcinoma↗

An ice nucleation active gene of Erwinia ananas. Sequence similarity to those of Pseudomonas species and regions required for ice nucleation activity.

The ice nucleation active gene, inaA, of Erwinia ananas IN-10 has been sequenced. This gene encodes a protein composed of 1322 amino acid residues. The inaA protein contains a 1120-residue segment consisting of 70 repeats of closely related 16 amino acid motifs (R-domain), which is flanked by N- and C-terminal sequences (N- and C-domains, respectively). Its primary structure is similar to, but not identical with, those of Pseudomonas inaW and inaZ gene products. By truncating the inaA gene to various extents, it was found that deletion of the C-domain resulted in complete loss of the ice nucleation activity, whereas removal of the N-domain led to a moderate decrease in the activity. Complete loss of the activity was also observed when the N-domain plus a large part of the P-domain were deleted. It is suggested that the C-domain is required for the assembly of inaA protein to form a functional ice nucleus.

Amino Acid Sequence↗

Calyculin A and okadaic acid: inhibitors of protein phosphatase activity.

Calyculin A and okadaic acid induce contraction in smooth muscle fibers. Okadaic acid is an inhibitor of phosphatase activity and the aims of this study were to determine if calyculin A also inhibits phosphatase and to screen effects of both compounds on various phosphatases. Neither compound inhibited acid or alkaline phosphatases, nor the phosphotyrosine protein phosphatase. Both compounds were potent inhibitors of the catalytic subunit of type-2A phosphatase, with IC50 values of 0.5 to 1 nM. With the catalytic subunit of protein phosphatase type-1, calyculin A was a more effective inhibitor than okadaic acid, IC50 values for calyculin A were about 2 nM and for okadaic acid between 60 and 500 nM. The endogenous phosphatase of smooth muscle myosin B was inhibited by both compounds with IC50 values of 0.3 to 0.7 nM and 15 to 70 nM, for calyculin A and okadaic acid, respectively. The partially purified catalytic subunit from myosin B had IC50 values of 0.7 and 200 nM for calyculin A and okadaic acid, respectively. The pattern of inhibition for the phosphatase in myosin B therefore is similar to that of the type-1 enzyme.

Alkaline Phosphatase↗

Monoclonal antibody raised against tetrodonic acid, a derivative of tetrodotoxin.

Tetrodonic acid, a relatively non-toxic derivative of tetrodotoxin, was conjugated with bovine serum albumin and injected i.p. to BALB/c mice. After several injections, spleen cells were isolated, fused with myeloma cells X63-Ag8-6.5.3. and cloned by the limiting dilution method. The monoclonal antibody produced in ascites fluid in the mouse by the cloned cell showed an increasing reactivity with tetrodotoxin at concentrations ranging from 0.03 to 100 micrograms per well.

Animals↗

Phosphorylation of paramyosin.

1. Myofibrils isolated from Mercenaria mercenaria were phosphorylated by endogenous kinase. Over a range of ionic strengths only paramyosin was phosphorylated. 2. Thiophosphorylation of paramyosin caused an inhibition of steady-state actin-activated ATPase activity of the myofibrils. 3. It is proposed that the endogenous kinase is the catalytic subunit of the cAMP-dependent protein kinase. 4. The sequence around the phosphorylation site was determined. 5. The phosphorylation site probably is close to the C-terminus of the paramyosin molecule.

Animals↗

Diagnostic value of QRST isointegral maps in detecting myocardial infarction complicated by bundle branch block.

The clinical usefulness of QRST isointegral maps (IQRST map) for detecting myocardial infarction that was complicated by intraventricular conduction disturbances was evaluated in patients with right bundle branch block (group RBBB, 64 patients) and left bundle branch block (group LBBB, 40 patients) by comparison with the normal mean IQRST map derived from 50 normal subjects. Myocardial infarction complicated the conduction disturbances in 24 of the 64 RBBB and in 18 of the 40 LBBB patients. A correlation coefficient was used for assessing the similarity of each map pattern with the normal mean IQRST map. The difference map was made by subtracting the average normal IQRST map from each abnormal IQRST map, and those differences that were less than 2 SD from the mean were retained as a significant area. The number of leads and their sum of differences were used to represent the size of the difference map. Correlation coefficients were significantly (p less than 0.001) smaller in patients with bundle branch block complicated by myocardial infarction than in patients with conduction disturbances not complicated by myocardial infarction. A significant area emerged in the difference map in all patients with myocardial infarction complicated by conduction disturbances. The emergence of a significant area revealed high diagnostic accuracy for detecting myocardial infarction in group RBBB (89.1%). The size of a significant area in a difference map was significantly larger in cases with complicated myocardial infarction than in cases with uncomplicated myocardial infarction in either group RBBB or group LBBB (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗

Calcium-independent activation of contractile apparatus in smooth muscle by calyculin-A.

Calyculin-A (CL-A), a novel marine toxin isolated from Discodermia calyx, caused contraction in the smooth muscle of guinea pig taenia ceci and rat aorta in the presence or absence (with 1 mM ethylene glycol bis(beta-aminoethyl ether)-N,N'-tetraacetic acid) of external Ca++ at concentrations ranging from 1 X 10(-8) to 1 X 10(-6) M. In the presence of external Ca++, the contraction induced by CL-A was accompanied by an increase in the cytosolic free Ca++ concentration [( Ca++]cyt) as measured by the fluorescence indicator fura-2. Verapamil (3 X 10(-6) M) inhibited the increase in [Ca++]cyt, but not tension development caused by CL-A. In the absence of external Ca++, CL-A still caused contraction without changing [Ca++]cyt. Thus, from studies with intact smooth muscle it was demonstrated that, in the absence of external Ca++, CL-A can induce a contraction that was not accompanied by an increase in [Ca++]cyt. In permeabilized taenia, CL-A caused contraction in the absence of Ca++ (with 2 mM ethylene glycol bis(beta-aminoethyl ether)N,N'-tetraacetic acid) at concentrations similar to those required to contract intact tissue. This contraction was inhibited by the nonselective kinase inhibitors such as amiloride (1 X 10(-3) M) and K-252a (2 X 10(-5) M). Low concentrations of Ca++ (approximately 1 X 10(-6) M) augmented the CL-A-induced contraction in the permeabilized taenia. In native actomyosin prepared from chicken gizzard CL-A induced phosphorylation of the 20 kDa myosin light chain (MLC) in the absence of Ca++.(ABSTRACT TRUNCATED AT 250 WORDS)

2',3'-Cyclic-Nucleotide Phosphodiesterases↗