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Biomedical subjects

J Fujii

Publications and source records attributed to J Fujii.

At least 199 records · Page 11Linked to original sources
PubMed1989

Slow/cardiac sarcoplasmic reticulum Ca2+-ATPase and phospholamban mRNAs are expressed in chronically stimulated rabbit fast-twitch muscle.

Fast-twitch extensor digitorum longus muscles of the rabbit were subjected to chronic low-frequency stimulation during different time periods. Changes in the relative amounts of mRNAs encoding fast and slow/cardiac Ca2+-ATPase isoforms were assessed through the use of an RNase-protection assay. Stimulation-induced increases in slow cardiac Ca2+-ATPase and phospholamban mRNAs were quantified by mRNA hybridization. Prolonged stimulation resulted in an exchange of the fast with the slow/cardiac Ca2+-ATPase isoform mRNAs. The exchange was complete after 72 d of stimulation as compared with normal slow-twitch soleus muscle. The tissue content of phospholamban mRNA reached levels similar to that found in normal slow-twitch soleus muscle by the same time. The conversion of the sarcoplasmic reticulum coincided with the fast-to-slow troponin C isoform transition, previously investigated in the same muscles.

Animals↗
PubMed1989

Expression and site-specific mutagenesis of phospholamban. Studies of residues involved in phosphorylation and pentamer formation.

Full-length cDNAs encoding either dog cardiac or rabbit skeletal muscle phospholamban were expressed transiently in COS-1 cells. The expressed protein displayed the mobility of a pentamer when dissolved in sodium dodecyl sulfate and separated in polyacrylamide gels, and of a monomer when boiled prior to polyacrylamide gel separation. Site-specific mutagenesis was used to analyze the roles of several amino acids in the structure and function of the protein. Ser16 and Thr17 were shown to be phosphorylated uniquely by cAMP- and calmodulin-dependent protein kinases, respectively, confirming earlier observations on the native protein (Simmerman, H. K. B., Collins, J. H., Theibert, J.L., Wegener, A.D., and Jones, L.R. (1986) J. Biol. Chem. 261, 13333-13341). Arg13 and Arg14 were shown to be essential for both types of phosphorylation, and Arg9 was shown to be essential for calmodulin-dependent phosphorylation. In studies of pentamer stability, mutation of Gln22-Gln23 to Ala-Ala or Glu-Glu, of Gln26-Asn27 to Glu-Asp, or of Gln29-Asn30 to Glu-Asp had no effect on thermal stability of the pentamer, suggesting that hydrogen bonding involving these residues in domain IB is not important for pentamer stability. By contrast, mutation of Cys36, Cys41, and Cys46 in transmembrane domain II to Ser, Ala, or Phe diminished the stability of the pentamer when microsomal proteins were dissociated in sodium dodecyl sulfate and separated by polyacrylamide gel electrophoresis. In particular, the Cys41 to Phe mutant existed as a monomer at ambient temperature. These results suggest that the intramembranous cysteine residues are important for pentamer formation even though they are not disulfide-bonded.

Amino Acid Sequence↗
PubMed1989

Functional consequences of alterations to amino acids located in the catalytic center (isoleucine 348 to threonine 357) and nucleotide-binding domain of the Ca2+-ATPase of sarcoplasmic reticulum.

The sequence of 10 amino acids (ICSDKTGTLT357) at the site of phosphorylation of the rabbit fast twitch muscle Ca2+-ATPase is highly conserved in the family of cation-transporting ATPases. We changed each of the residues flanking Asp351, Lys352, and Thr353 to an amino acid differing in size or polarity and assayed the mutant for Ca2+ transport activity and autophosphorylation with ATP or P1. We found that conservative changes (Ile----Leu, Thr----Ser, Gly----Ala) or the alteration of Cys349 to alanine did not destroy Ca2+ transport activity or phosphoenzyme formation, whereas nonconservative changes (Ile----Thr, Leu----Ser) did disrupt function. These results indicate that very conservative changes in the amino acids flanking Asp351, Lys352, and Thr353 can be accommodated. A number of mutations were also introduced into amino acids predicted to be involved in nucleotide binding, in particular those in the conserved sequences KGAPE519, RDAGIRVIMITGDNK629, and KK713. Our results indicate that amino acids KGAPE519, Arg615, Gly618, Arg620, and Lys712-Lys713 are not essential for nucleotide binding, although changes to Lys515 diminished Ca2+ transport activity but not phosphoenzyme formation. Changes of Gly626 and Asp627 abolished phosphoenzyme formation with both ATP and Pi, indicating that these residues may contribute to the conformation of the catalytic center.

Amino Acid Sequence↗
PubMed1989

Hydroxyproline in all parts of the rabbit heart in hypertension and in its reversal.

To investigate the response of cardiac collagen to arterial hypertension and its reversal, we measured the hydroxyproline concentration in all parts of the rabbit heart in three groups: a chronic hypertension group (n = 11), a reversal group (n = 9) and an age-matched control group (n = 10). Left ventricular overload was produced by one-kidney, one-clip Goldblatt hypertension and its reversal was produced by clip release. The wet weight of every part of the left side of the heart was increased by hypertension and all parts except for the papillary muscles showed regression with reversal. However, the response of collagen was quite different among the various parts of the heart. The wet weights of the left atrium, left ventricular free wall, interventricular septum, and anterior and posterior papillary muscle in the hypertension group were 144 (P less than 0.01), 140 (P less than 0.01), 134 (P less than 0.01), 144 (P less than 0.01) and 129% (P less than 0.01), respectively, and those in the reversal group were 92, 113 (P less than 0.05), 107 (P less than 0.05), 142 (P less than 0.05) and 126% (P less than 0.05) of those in the control group, respectively. Hydroxyproline concentrations in the respective parts in the hypertension group were 79 (P less than 0.05), 91, 98, 121 (P less than 0.05) and 120% (P less than 0.05) and those in the reversal group were 86, 93, 94, 107 and 110% of those in the control group.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Regulation of myocardial Ca2+-ATPase and phospholamban mRNA expression in response to pressure overload and thyroid hormone.

The sarcoplasmic reticulum (SR) and the contractile protein myosin play an important role in myocardial performance. Both of these systems exhibit plasticity--i.e., quantitative and/or qualitative reorganization during development and in response to stress. Recent studies indicate that SR Ca2+ uptake function is altered in adaptive cardiac hypertrophy and failure. The molecular basis (genetic and phenotypic) for these changes is not understood. In an effort to determine the underlying causes of these changes, we characterized the rabbit cardiac Ca2+-ATPase phenotype by molecular cloning and ribonuclease A mapping analysis. Our results show that the heart muscle expresses only the slow-twitch SR Ca2+-ATPase isoform. Second, we quantitated the steady-state mRNA levels of two major SR Ca2+ regulatory proteins, the Ca2+-ATPase and phospholamban, to see whether changes in mRNA content might provide insight into the basis for functional modification in the SR of hypertrophied hearts. In response to pressure overload hypertrophy, the relative level of the slow-twitch/cardiac SR Ca2+-ATPase mRNA was decreased to 34% of control at 1 week. The relative Ca2+-ATPase mRNA level increased to 167% of control after 3 days of treatment with thyroid hormone. In contrast, in hypothyroid animals, the relative Ca2+-ATPase mRNA level decreased to 51% of control at 2 weeks. The relative level of phospholamban mRNA was decreased to 36% in 1-week pressure overload. Hyperthyroidism induced a decrease to 61% in the phospholamban mRNA level after 3 days of treatment, while hypothyroidism had virtually no effect on phospholamban mRNA levels. These data indicate that the expression of SR Ca2+-ATPase and phospholamban mRNA may not be coordinately regulated during myocardial adaptation to different physiological conditions.

Animals↗
PubMed1989

Age-related reduction in the number of rabbit erythrocyte Na, K-ATPase.

The number of Na, K-ATPase units, assessed by [3H]-ouabain binding assay, was significantly less in erythrocytes from 17 mature (30-week) rabbits than in those from 17 young (16-week) ones (0.179 +/- 0.010 vs. 0.263 +/- 0.014 pmol/10(9) cells, p less than 0.01). Consistent with this finding, the erythrocyte sodium concentration was significantly higher in mature rabbits as compared with young animals (17.1 +/- 1.30 vs. 10.8 +/- 0.68 mmol/liter, p less than 0.01). When the assays were repeated in 8 of the young rabbits 8 weeks after the initial determination, both variables were found to be similar to the levels from mature animals. The data suggest that the activity of erythrocyte sodium pump declines with aging or growth of donor rabbits, as a result of a reduction in the number of Na, K-ATPase units.

Aging↗
PubMed1989

[A clinical study of amino acid metabolism in cancerous hypoproteinemia--the role of branched chain amino acids (BCAA) and appropriate compositions of BCAA in parenteral nutrition].

In order to determine the nutritional effects of BCAA compositions in the treatment of cancerous hypoproteinemia, the appropriate ratio of I-leu: Leu: Val and the proportion of BCAA to Total Amino Acids were investigated. As for results, indices such as the serum albumin, the RBP and N-balance quickly recovered to normal levels when the ratio of I-leu: Leu: Val was 1.0:1.8:1.0 and the proportion o BCAA to TAA was 31%. These composition thus may be suitable for the treatment of cancerous hypoproteinemia.

Amino Acids↗
PubMed1989

[Production of experimental infective endocarditis in rabbit mitral valve prolapse induced by vagus stimulation].

An experimental model of bacterial endocarditis was developed in rabbits with mitral valve prolapse which was induced by vagus stimulation. Endocarditis was produced by injecting bacteria including pseudomonas pseudoalkaligenes (pathogenic for rabbits) and streptococcus viridans taken from human beings. Both bacteria induced bacterial endocarditis with high incidence, which was augmented by higher dose or earlier administration of bacterial after producing mitral valve prolapse. Similarly, verruca was produced more frequently by higher dose of bacteria or earlier administration of bacteria. The present data are similar to those of human beings, so that it is helpful to a study of bacterial endocarditis in a clinical setting.

Animals↗
PubMed1988

HTLV-II transactivation is regulated by the overlapping tax/rex nonstructural genes.

The human T-cell leukemia virus (HTLV) types I and II have two nonstructural genes that are encoded in overlapping reading frames. One of these genes, known as tax, has been shown to encode a protein responsible for enhanced transcription (transactivation) from the viral long terminal repeats (LTRs). Genetic evidence indicates that the second nonstructural gene of HTLV-II, here designated rex, acts in trans to modulate tax gene-mediated transactivation in a concentration-dependent fashion. The rex gene may regulate the process of transactivation during the viral life cycle.

Base Sequence↗
PubMed1988

Rabbit cardiac and slow-twitch muscle express the same phospholamban gene.

The nucleotide sequences of cDNAs encoding phospholamban were found to be virtually identical when the cDNA clones were isolated from rabbit slow-twitch (soleus) and rabbit cardiac muscle libraries. These findings demonstrate that both types of muscle express the same phospholamban gene. The deduced amino acid sequences of rabbit and dog phospholamban were identical except for a change from Asp (dog) to Glu (rabbit) at position 2. The nucleotide sequences of the 5'- and the very long 3'-untranslated regions of rabbit and dog phospholamban cDNAs also exhibited a high percentage of identity.

Amino Acid Sequence↗
PubMed1988

Tumor markers in pleural effusion diagnosis.

In order to discriminate between malignant and benign effusions, the values of carcinoembryonic antigen (CEA), ferritin, beta2-microglobulin (BMG), acid-soluble glycoprotein (ASP), tissue polypeptide antigen (TPA), adenosine deaminase (ADA), and immunosuppressive acidic protein (IAP) were measured in the pleural fluid of 54 patients with lung cancer, 20 with malignancies other than lung cancer, 18 with tuberculous pleurisy, and 22 with benign diseases other than tuberculosis. CEA levels in malignant effusions were significantly higher than those in benign effusions. At a cutoff level of 5 ng/ml, 68% of the patients with lung cancer and 44% of the patients with other malignancies showed elevated pleural fluid CEA levels. In 13 lung cancer cases with negative pleural fluid cytology, nine cases had elevated pleural fluid CEA levels. The mean pleural fluid BMG level of patients with benign diseases was significantly higher than that of patients with malignant diseases, but there was a marked overlap between those with malignant and benign diseases. No significant differences were found in the pleural fluid ferritin, ASP, TPA, and IAP levels between malignant and benign conditions. ASP and IAP pleural fluid levels showed significant correlations with the pleural fluid C-reactive protein (CRP) concentrations suggesting that they also reflect inflammatory activity. The mean ADA activity in tuberculous effusion was significantly higher than that resulting from other causes of pleural effusion.

Adult↗
PubMed1988

ST-segment fluctuation during treadmill exercise in patients with angina pectoris.

The level of the ST-segment fluctuates transiently during treadmill exercise in some patients with angina pectoris. In the present study, the incidence and clinical significance of ST-segment fluctuation were studied before and after propranolol in 52 patients with angina pectoris. A transient greater than 0.5-mm (0.05 mV) upward shift of the ST-segment during a graded treadmill test was considered a significant fluctuation in leads without signs of previous myocardial infarction. The fluctuation was observed in three of 30 patients with rest or rest and effort angina pectoris before propranolol and in 14 of them after propranolol, while only one of 22 patients with effort angina alone showed fluctuation after the drug. Coronary arteriography revealed that in 15 patients showing ST-segment fluctuation with propranolol, seven patients had no significant coronary stenosis, six had one-vessel disease and two had two-vessel disease. In 24 patients with documented coronary artery spasm, ST-segment fluctuation was induced in two (8%) before propranolol and in 13 (54%) after propranolol. Our results suggest that ST-segment fluctuation during graded treadmill exercise may be related to transient coronary vasospasms exacerbated by propranolol.

Adult↗