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Biomedical subjects

Y Nabeshima

Publications and source records attributed to Y Nabeshima.

At least 91 records · Page 5Linked to original sources
PubMed1993

MyoD and myogenin act on the chicken myosin light-chain 1 gene as distinct transcriptional factors.

Expression of MyoD, myogenin, MRF4, and Myf-5 converts nonmuscle cells to muscle cells. In an attempt to analyze the roles of these factors, we have investigated their effects on transcription driven by the promoter of the chicken myosin alkaline light-chain (MLC1) gene. The activation by CMD1 or c-myogenin (chicken MyoD or myogenin, respectively) was dependent on the existence of a muscle-specific regulatory region located from positions -2096 to -1743. Its distal half, containing a pair of E boxes (CANNTG), had been previously characterized as an enhancer responsive to CMD1 but not to c-myogenin. In this study, we report the identification of another enhancer in the muscle-specific regulatory region which is preferentially responsive to c-myogenin. Deletion and mutation analyses indicated that this enhancer requires a single E box and its flanking sequences. Furthermore, analysis of chimeric proteins of CMD1 and c-myogenin indicated that regions outside the basic helix-loop-helix domain of c-myogenin are involved in the specificity of the enhancer. These results show that CMD1 and c-myogenin act on the MLC1 gene by recognizing different upstream DNA sequences and that direct or indirect interactions between the regions outside the basic helix-loop-helix domain and flanking sequences of E boxes are involved in the target sequence specificity.

Animals↗
PubMed1993

Controlled release of proteins from 2-hydroxyethyl methacrylate copolymer gels.

A series of hydrogels with large pores was synthesized by the precipitation polymerization of 2-hydroxyethyl methacrylate (HEMA) with crosslinking agent in aqueous solution. Such gels are potentially useful for the controlled release of large-molecular-weight species such as proteins. In this study, the release behavior of lysozyme and alpha-amylase from hydrogels formed from HEMA or HEMA with a comonomer was studied. It was found that the polymer composition affected the total amount of lysozyme released and its activity. Effects were smaller with alpha-amylase. Charged gels, containing a phosphate moiety, released larger amounts of lysozyme at a reduced rate as a result of charge-charge interactions.

Biocompatible Materials↗
PubMed1993

[Production of transgenic mice overexpressing rabbit Na+/H+ antiporter].

Recent investigations have raised a possibility that abnormal ion transportation through cell membranes may be involved in the pathogenesis of essential hypertension. In order to test the hypothesis that increased activity of Na+/H+ antiporter may cause hypertension, we developed transgenic mice overexpressing the Na+/H+ antiporter. We isolated a full-length cDNA clone encoding the rabbit Na+/H+ antiporter and constructed the transgene by ligating it with the human elongation factor 1 alpha promoter. We obtained three transgenic strains which express the transgene in various tissues such as kidney, heart and aorta. These transgenic mice may be useful for the analysis of pathogenesis of essential hypertension.

Animals↗
PubMed1993

[Fetal heart rate monitoring during maternal swimming].

Fetal physiology during swimming in pregnancy has remained unclear. To assess the fetal status, the fetal heart rate (FHR) was measured during maternal swimming. We studied individually a group of seventeen women in normal pregnancy (during their third trimester, 35-38th week) who participated in a maternal swimming class. We adapted the Doppler ultra-sound transducer for underwater use and attached it to the mothers' abdomen. FHR was recorded before, during and after swimming. The group swam from 375 to 750 meters in 33 to 41 minutes. During swimming, motion artifacts interfered with the FHR signal and we were only able to detect FHR in eleven in the group. The mean FHR rose significantly during swimming in eight of eleven women compared to the FHR recorded before the exercise. The FHR pattern was reactive and baseline variability was preserved in all cases. No pathological deceleration was seen. It is concluded that maternal swimming under such class conditions is safe for the fetus.

Adult↗
PubMed1993

Analysis of Bankart lesion in the recurrent dislocation or subluxation of the shoulder.

The Bankart repair was used to restore anterior stability to 50 patients (51 shoulders) with recurrent dislocation or subluxation of the shoulder. The patients had preoperative diagnostic testing to determine the presence and extent of a Bankart lesion. At surgery, the Bankart lesion was carefully observed for its location, and extent of involvement of associated structures. The intraoperative findings of the severity of the Bankart lesion were compared with the preoperative data. The glenoidal fossa was considered like a clock, and the location and severity of the Bankart lesion was expressed by the hour of the clock. The Bankart lesion most frequently occurred from two o'clock to six o'clock in the right shoulder, and from six o'clock to ten o'clock in the left shoulder. The region of the glenohumeral joint at highest risk of exhibiting pathology after recurrent dislocation or subluxation of the shoulder was usually located from the central anterior edge of the glenohumeral joint to the inferior margin. Arthrogram with computed tomography was the most useful preoperative diagnostic test to demonstrate the Bankart lesion. The greatest number of inferior postoperative results were observed in patients with Bankart lesions of large size.

Adult↗
PubMed1992

Differential trans-activation of muscle-specific regulatory elements including the mysosin light chain box by chicken MyoD, myogenin, and MRF4.

We have isolated cDNAs encoding a chicken homologue of MRF4 (cMRF4) in addition to chicken MyoD (CMD1) and myogenin (c-myogenin) described previously. In an attempt to understand the roles that cMRF4, CMD1, and c-myogenin play in chicken myogenesis, the effects of these factors on muscle-specific cis-elements identified in regulatory regions of myosin alkali light chain (MLC) genes were examined. The promoter analysis of some of MLC genes has revealed two sorts of muscle-specific positive regulatory elements to date, an enhancer located upstream of the adult type LC1 gene and a cis-element, termed an MLC box, conserved among promoters of various MLC genes. The LC1 enhancer was exclusively trans-activated by CMD1. Although c-myogenin also activated transcription driven by the LC1 promoter, it was suggested that c-myogenin requires a cis-element(s) other than the CMD1-responsive enhancer. Chicken MRF4 could not trans-activate any of the constructs containing the LC1 promoter. In contrast, the promoter of the embryonic L23 gene was trans-activated by all of the three factors. From deletion and mutation analysis, the MLC box was shown to be involved in their positive regulation. These results extend previous observations that individual myogenic regulatory factors exhibit different capabilities in transcriptional activation of muscle-specific genes by acting distinctively upon their regulatory elements.

Amino Acid Sequence↗
PubMed1992

Cloning of the Drosophila prospero gene and its expression in ganglion mother cells.

The Drosophila central nervous system comprises an enormous diversity of neurons that are originated from neuronal stem cells, neuroblasts. They generate a specific series of ganglion mother cells, each of which is once cleaved into a pair of neurons. Among genes known to control neurogenesis, prospero (pros) was recently identified as a gene required for gene expression specifying properties of some identified neurons. Here we report that pros encodes a nuclear protein containing a homeodomain-like sequence. In neuronal lineages of the central nervous system, pros protein is specifically detected in ganglion mother cells, although their parental neuroblasts have begun expressing a significant level of pros transcripts, suggesting a post-transcriptional control of pros expression. Our results provoke that in neuronal cell differentiation ganglion mother cells might play a pivotal role associating with the pros function.

Amino Acid Sequence↗
PubMed1992

Positive and negative gene regulation in muscle.

By the analysis of cis and trans-acting element involved in transcriptional regulation of chicken myosin alkali light chain genes, we have identified the MLC box, muscle specific enhancer element and negative regulatory element. The MLC box is an essential element for the expression of MLC genes located at approximately 100 bp upstream from mRNA start sites. The core sequence of MLC box is similar to the consensus of actin gene CArG box and SRE of c-fos oncogene. In vitro DNA-protein binding assay has revealed that the MLC box, CArG box and SRE might bind to a common or a similar protein complex. CMD1, cMyogenin, and cMRF4 transactivate the promorter with an intact MLC box, but not the promoter lacking MLC box, indicating that the MLC box itself is transactivated by the myogenic regulatory factors. This transactivation must have been due to the indirect effect of the myogenic regulatory factors, because chicken myogenic factors do not bind to the MLC box. A cis element identified at about 150 bp upstream from the cap site of cardiac MLC gene suppresses the cardiac MLC gene expression in skeletal muscle cells but not in cardiac muscle cells. The protein(s) bound to NRE might be identical with one of proteins bound to SRE. NRE may block the function of MLC box and resultantly inhibits the expression of cardiac MLC1 gene in skeletal muscle cells. Skeletal muscle enhancer at -2 kb of skeletal MLC1f gene is composed of two subelements P and D, cooperative action between them is required for sufficient enhancer activity. CMD1 and myogenin bind to the enhancer sequences of skeletal MLC1 gene and MCK gene and transactivate these genes preferentially in skeletal muscle cells. In addition to the CMD1 responsible enhancer, another cis-element is required for transactivation of the MLC1f gene by cMyogenin. An E-box adjacent to MLC box may co-work with the enhancer to increase the expression of MLC1f gene. Muscle specific and developmentally regulated expression of MLC gene family is regulated by the combination of these cis and trans-acting elements.

Animals↗
PubMed1992

[Effect of maternal exercise with graded treadmill on fetal heart rate].

The effect of maternal exercise on the fetal and maternal heart rate was studied in 10 normal pregnant women in their third trimester. They participated in 15 minute graded treadmill exercise. Fetal heart rate (FHR), uterine contraction, maternal blood pressure, electrocardiogram, and oxygen consumption were monitored, before, during, and after exercise. FHR could be monitored in 7 women. Maternal maximal heart rate ranged from 143 to 168 bpm which approximated 61% to 84% training intensity. FHR showed a significant rise in 4 women during exercise. Of four cases which showed more than 70% training intensity, 3 demonstrated fetal tachycardia over 160 bpm and one demonstrated fetal bradycardia under 120 bpm. The baseline fetal heart rate remained from 120 bpm to 160 bpm in cases under 70% training intensity. There was no pathological deceleration and the variability was preserved in all cases. It is suggested that maternal exercise intensity should be less than 70% of the patient's maximal capacity, which approximates maternal heart rate of about 150 bpm.

Adult↗
PubMed1991

A stable cellular marker for the analysis of mouse chimeras: the bacterial chloramphenicol acetyltransferase gene driven by the human elongation factor 1 alpha promoter.

We have developed a method of marking of mouse cells by means of transfection of a foreign gene. The transgene chosen here was the plasmid pEF321CAT which contains the bacterial chloramphenicol acetyl transferase (CAT) gene linked to the promoter region of the human polypeptide chain elongation factor 1 alpha (hEF1 alpha) gene. Evaluation of the plasmid pEF321CAT as a cellular marker for mouse cells involved intensive examination of a transgenic mouse carrying pEF321CAT. The CAT gene was expressed in all tissues examined, demonstrating that the hEF1 alpha promoter was active in a wide range of mouse cells. The plasmid itself did not exert any harmful effect on the normal development of mice, and the CAT activity was immunohistologically detectable on sectioned tissues by the use of anti-CAT serum. When the plasmid was transferred into embryonal carcinoma (EC) cells and embryonic stem (ES) cells, the CAT gene was also found to be expressed constantly irrespective of their differentiation. These results demonstrated that the plasmid pEF321CAT can be used as a reliable and feasible cellular marker that would distinguish unequivocally the cells of each of genotype in chimeric tissues.

Aging↗
PubMed1991

Cytotoxic activity of marine algae and a cytotoxic principle of the brown alga Sargassum tortile.

Partition fractions of hexane, CCl4 and CHCl3 from methanolic extracts of marine algae were each examined for cytotoxic activities against cultured P-388 lymphocytic leukemia cells. Cytotoxic activities were found for partition fractions of 21 species of seaweed. Bioactivity-guided fractionation of the CCl4 partition fraction from Sargassum tortile, exhibiting the most prominent activity, afforded dihydroxysargaquinone (1) and sargatriol (2) previously isolated from this alga. The former was evaluated as a cytotoxic principle, and the latter, showing moderate activity, was suggested to be an artifact derived from 1 during the isolation procedure.

Animals↗
PubMed1991

[Pulsatile release of gonadotropins in athletic women].

In athletic women menstrual disorders such as luteal insufficiency, oligomenorrhea and amenorrhea are often seen. It has been suggested that such disorders may be related to hypothalamic-pituitary axis insufficiency caused by physical activity. To investigate the mechanism by which the disorder is promoted, episodic gonadotropin secretion was studied in 10 athletic women (normal ovulatory, 5; short luteal, 5) and 6 non-athletic controls. In the early follicular phase of the menstrual cycle, blood samples were obtained through an indwelling venous catheter at 15-minute intervals for 4 hours. The concentrations of luteinizing hormone (LH) and follicle-stimulating hormones (FSH) were measured by radioimmunoassay. The mean LH and FSH concentrations in the athletic groups were lower than those in the control group. LH pulse frequencies in the short luteal group were 2.8 +/- 0.2 times/four hours (mean +/- standard error) and decreased compared with that of the control group (3.7 +/- 0.2 times/four hours; p less than 0.001). The LH pulse amplitudes of all three groups were similar. Pulsatile FSH secretion was also observed in all the women. Dynamism of FSH was the same as that of LH except for low pulse amplitude in the athletic groups. These data suggest that hypothalamic-pituitary axis insufficiency, especially of hypothalamic origin may be caused by athletic activity.

Adult↗
PubMed1990

Myogenin contains two domains conserved among myogenic factors.

Previously, three structurally related proteins, MyoD, myogenin, and Myf5, have been identified, and each of them was found to convert C3H10T1/2 fibroblasts to myoblasts when their respective cDNAs were expressed under the control of a viral promoter. Here, we describe the cloning and DNA sequencing of myogenin cDNAs from chicken and mouse. They encode polypeptides highly homologous to each other, but the polypeptide sequences we have obtained are not homologous in the carboxyl-terminal 70 amino acids with those previously reported for mouse, rat, and human because of a single base deletion in the previously reported cDNAs. Determination of genomic sequence coding for myogenin revealed that the mouse myogenin cDNA presented here corresponds to a correct transcript of the gene, and the nucleotide sequence of the previously reported cDNA is incorrect. A comparison of chicken and mouse myogenins with other myogenic regulatory factors, MyoD and Myf-5, identified a domain with an interesting feature located in the carboxyl terminus of these proteins in addition to the myc homology domain previously reported.

Amino Acid Sequence↗
PubMed1990

Regulation of the chicken embryonic myosin light-chain (L23) gene: existence of a common regulatory element shared by myosin alkali light-chain genes.

The transcriptional regulation of the chicken myosin alkali light-chain (MLC) L23 gene was analyzed. Two different types of cis-regulatory regions were identified: one was a silencerlike region located between 3.7 and 2.7 kilobases upstream of the mRNA initiation site, and the other was essential for the expression of L23 in skeletal muscle cells and was located between 106 and 91 base pairs upstream of the cap site. This 16-base-pair cis-acting element was designated as the MLC box since it is well conserved in various muscle-specific MLC promoter regions. The activity of the MLC box showed tissue specificity. To analyze the relationship between the nucleotide sequence and the activity of the MLC box precisely, mutation analysis was performed. The 16-base-pair sequence was indispensable for the active transcription of L23 gene, and the MLC box could function in either orientation. The inverted sequence of the MLC box was similar to the sequence of the alpha-actin CArG box. By using a gel mobility retardation assay, the nuclear protein(s) that binds to both MLC box and CArG box was detected with nuclear extract prepared from chicken embryonic breast muscle. These observations imply that a common factor regulates the coordinate expression of these contractile proteins in muscle differentiation.

Animals↗
PubMed1989

Separation of B and T lymphocytes by a hybrid field-flow fractionation/adhesion chromatography technique.

A hybrid of the techniques of cellular adhesion chromatography and field-flow fractionation has been used for the effective separation of rat mesenteric B and T lymphocytes with nearly complete recovery of both cell species. Use of this hybrid technique also allows the relative binding strengths of cells to biomaterials and other surfaces to be rapidly and simply estimated. For the copolymer surface used here, B cells appear to bind with a force five times greater than T cells.

Animals↗
PubMed1989

Structural control of poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymers for differential retention of rat lymphocyte subpopulations.

Poly(2-hydroxyethyl methacrylate)-graft-polyamine copolymers (HA copolymers) of varying composition and chain length of the polyamine graft were prepared by radical copolymerization of 2-hydroxyethyl methacrylate with a given quantity of polyamine macromonomer having a controlled molecular weight, and their interaction with lymphocyte subpopulations (B and T cells) was estimated using a column method. From these results, it was revealed that the most suitable molecular structure of HA copolymers for separating lymphocyte subpopulations is that of an HA copolymer with 13 wt% of polyamine graft with a chain length 3000-6600 in molecular weight. It is concluded that the retention process of lymphocytes on the HA graft copolymers is driven primarily by ionic interactions between the protonated amino groups and the cells. However, the mode of the polyamine microdomain structure, that is the distribution of protonated amino groups, and the conformation of the polyamine chains, which varies with the chain length of the HA copolymer, are also important factors in determining differential lymphocyte retention.

Animals↗
PubMed1989

Effect of naloxone on hormonal changes during exercise.

It is well established that prolactin release during exercise is one of the important factors in exercise-induced menstrual dysfunction. The purpose of this study is to clarify the mechanisms of prolactin release during exercise. Ten female athletes measured their BBT every morning. They performed incremental exercise on a cycle ergometer, with or without naloxone, on the 5th to 8th days of the follicular phase. Three minutes before the exercise, 0.4mg of naloxone was injected intravenously and a further 1.6mg/hr of naloxone was continuously infused during exercise. Blood samples were collected after 60 minutes bed rest (Rest), at the time when the heart rates reached 150 bpm (Submax), the point of exhaustion (Max) during exercise and after 60 minutes bed rest following exercise (After 1hr). The levels of prolactin in serum, dopamine, beta-endorphin. VIP and ACTH in the plasma were measured. Whereas prolactin increased significantly at Submax (p less than 0.05) and Max (p less than 0.001), the increase in prolactin was suppressed by the administration of naloxone (p less than 0.05). Dopamine showed no remarkable change during exercise, with or without naloxone. There were significant increases in beta-endorphin at Max (p less than 0.001), VIP at Submax and Max (p less than 0.001), but these increases were suppressed by the administration of naloxone (p less than 0.001). ACTH which had markedly increased at Submax (p less than 0.025) and Max (p less than 0.001) showed a slight tendency to decrease following the administration of naloxone, but there were no significant differences in both groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗