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Biomedical subjects

M J Smith

Publications and source records attributed to M J Smith.

At least 253 records · Page 14Linked to original sources
PubMed1989

Multiple zones in the sequence of calreticulin (CRP55, calregulin, HACBP), a major calcium binding ER/SR protein.

The complete amino acid sequence of CRP55, the major 55 kd calcium binding protein of the ER lumen, was deduced from the murine cDNA nucleotide sequence. This was completed using a novel application of PCR amplification. The mature 399 residue protein encoded is preceded by a 17 amino acid leader sequence and ends in the ER signal sequence, KDEL. The protein contains no calcium binding motifs of the EF hand type or of the form seen in calelectrin-related proteins. The major region of potential low affinity calcium binding sites is a polyacidic stretch towards the C terminus. The primary structure of the protein is markedly zonal. The N-terminal region, of approximately neutral net charge and hydrophobicity, is followed by a central proline-rich zone with repeat sequences separated from the polyacidic C-terminal stretch by a short hydrophobic sequence. The general shape suggested is a globular domain attached to an extended tail. Immunofluorescence studies show that the protein is present in skeletal muscle and indicate that it is a sarcoplasmic reticulum protein. We propose that the protein be named calreticulin to reflect its calcium binding activity and location in the ER and SR.

Amino Acid Sequence↗
PubMed1989

The genomic nucleotide sequences of two differentially expressed actin-coding genes from the sea star Pisaster ochraceus.

The genomic sequences of two differentially expressed actin genes from the sea star Pisaster ochraceus are reported. The cytoplasmic actin gene (Cy) is expressed in eggs and early development. The muscle actin gene (M) is expressed in tube feet and testes. Both genes contain an 1125-nucleotide coding region interrupted by three introns at codons 41, 121 and 204. Gene M contains two additional introns at codons 150 and 267. The intron position at codon 150, although present in higher vertebrate actins, has not been reported in actin genes from invertebrates. The M gene coding region has 89.5% nucleotide homology to the Cy gene, and differs from the Cy actin gene in 13 of 375 amino acids (aa), 11 of which are found in the C-terminal half of the gene. The C-terminal half of the M gene contains a significant number of muscle isotype codons. Even though there is only 1 aa change in the first 150 codons, there have been limited substitutions at many four-fold degenerate sites which may indicate selection pressure upon the secondary structure of the mRNA and/or a biased codon usage. Variant CCAAT, TATA, and poly(A)-addition signals have been identified in the 5' and 3' flanking regions. The presence of 5' and 3' splice junction sequences in the 5' flanking region of the Cy gene suggests the potential for an intron there.

Actins↗
PubMed1989

Gene arrangement in sea star mitochondrial DNA demonstrates a major inversion event during echinoderm evolution.

The mitochondrial (mt) DNA from the sea star Pisaster ochraceus has been isolated, restriction-mapped, and cloned into plasmid vectors. Both ribosomal RNA genes, the genes for 12 of the 13 mitochondrial proteins, and 11 of the tRNA genes have been localized by DNA sequence analyses. The sequence arrangement of the genes is markedly different from that seen in sea urchin mitochondrial DNA. A segment of the DNA molecule extending from tRNA(pro), including the tRNA cluster, ND1, ND2, and 16S genes, is inverted in relation to the sea urchin genome. The resulting gene order in the sea star is 12S, 16S, ND2, tRNA cluster, COI. As a result of the inversion, the transcriptional polarity of ND1, ND2, and 16S genes are opposite to that of the 12S and COI genes. The arrangement and transcriptional polarity of the other genes mapped here is the same as seen in urchin.

Animals↗
PubMed1989

Distribution of tunichrome and vanadium in sea squirt blood cells sorted by flow cytometry.

Specialized blood cells of many tunicates accumulate high concentrations of vanadium and phenolic peptide pigments called tunichromes (TC). In order to determine whether V and TC reside in the same cells, Ascidia nigra and Ascidia ceratodes blood cell subpopulations were isolated by fluorescence-activated cell sorting (flow cytometry) and chemically analyzed. V was found in the spherical, green/grey signet ring cells, and to a lesser degree in the mulberry-shaped, yellow/green morula cells (MRs), whereas free TC was detected mainly in MRs.

Animals↗
PubMed1989

Molecular analyses of gene expression during sea star spermatogenesis.

We have investigated actin gene expression during the annual spermatogenic cycle of Pisaster ochraceus by Northern blot analyses of testes RNAs pooled from defined spermatogenic stages. Specific probes for cytoplasmic (Cy) and muscle (M) actin gene products detect 2.3 and 2.1 kb transcripts, respectively. In addition, actin-coding sequence probes detect a third, much larger (3.5 kb) transcript designated FAT. Preliminary sequence analyses of two cDNAs representing portions of the FAT transcript show over 90% homology to Pisaster Cy actin at the amino acid level but only 80% nucleotide identity. The expression patterns of these three transcripts, plus two spermiogenic indicator transcripts (H3 histone and beta-tubulin), were determined over the cycle. The Cy transcript is seen at all stages but is ten- to 100-fold higher early in the cycle when mitotic activity predominates. The M transcript appears at the onset of gonadal growth and is maintained at constant levels through spermatogenesis consistent with the expansion of the muscular sheath surrounding the testes. The FAT, H3 histone, and beta-tubulin transcripts reach their highest levels in ripe testes when spermiogenic activity is maximal. The homology of the FAT transcript to actin, and its pattern of expression, suggest the hypothesis that this transcript may encode acrosomal actin.

Actins↗
PubMed1989

Conserved tRNA gene cluster in starfish mitochondrial DNA.

Partial sequencing of mtDNA from four long-diverged species of starfish reveals the existence of a conserved cluster of 13 tRNA genes, organized in a manner similar to that of the tRNA cluster of sea urchin mtDNA, but located at a position distant from the presumed replication origin. These findings suggest that a clustered organization of tRNA genes may have been present in the ancestral mitochondrial genome, and raise the possibility that tRNAs may have catalyzed the dispersal rather than the accumulation of the genes which encode them.

Amino Acid Sequence↗
PubMed1989

Sustained increases in plasma epinephrine concentration do not modulate renin release.

We examined the relationship between plasma renin activity (PRA) and renal arterial pressure (RAP) during 1) control conditions, 2) acute, and 3) chronic intravenous epinephrine (EPI) infusion (125 ng.kg-1.min-1). In eight conscious uninephrectomized dogs maintained on a normal sodium intake, the renin stimulus-response curve (RSRC) was determined by a stepwise reduction in RAP with an inflatable occluder around the renal artery controlled by a servo unit. The RSRC could be approximated by two lines intersecting at a threshold pressure (approximately 20 mmHg below control RAP). In the high-pressure range, PRA was relatively insensitive to changes in RAP, whereas, below threshold pressure, changes in RAP had large effects on PRA. During acute EPI infusion there was approximately a 40% increase in heart rate (control = 57 +/- 3 beats/min) and hematocrit (control = 30 +/- 1%) in association with a rise in plasma EPI concentration from 73 +/- 16 to 1,413 +/- 100 pg/ml; mean arterial pressure (MAP) was unchanged (94 +/- 3 mmHg). Moreover, EPI acutely increased basal PRA from 0.3 +/- 0.1 to 0.8 +/- 0.3 ng angiotensin I.ml-1.h-1 and shifted the RSRC to the right (increasing threshold pressure 7 mmHg) without altering the slope of the RSRC curve either above or below threshold pressure. In contrast, although plasma EPI concentration and hematocrit remained elevated during chronic EPI infusion, heart rate and basal PRA returned to preinfusion values. In addition, there were no significant long-term changes in MAP or in any of the parameters of the RSRC.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1989

Downregulation of beta-adrenergic receptor-mediated function during sodium restriction in humans.

Mononuclear leukocyte (MNL) beta 2-adrenergic receptor (beta 2-AR) binding and its linked adenylate cyclase sensitivity to isoproterenol were measured in nine healthy humans prior to and after 7 days of dietary sodium restriction to determine whether chronic physiological increases in plasma norepinephrine (NE) are associated with the downregulation of beta-AR-mediated function. Sodium restriction resulted in an increase in the plasma NE concentration (P less than 0.02) and decreases in MNL beta 2-AR density (P less than 0.001), affinity for antagonist (P less than 0.001), and adenylate cyclase sensitivity to isoproterenol (ANOVA, P less than 0.01). To determine whether this downregulation of MNL beta 2-AR-mediated function is related to the increased plasma NE concentration or to increased extravascular NE release, NE kinetics was assessed using compartmental analysis in each subject prior to and after sodium restriction. Sodium restriction caused a decrease in the plasma NE metabolic clearance rate (P less than 0.005) and in the volume of distribution of NE in the intravascular compartment (P less than 0.005), whereas the extravascular NE release rate was unchanged. Our data suggest that the downregulation of MNL beta 2-AR-mediated function in humans during dietary sodium restriction is a response to the increase in plasma NE.

Adenylyl Cyclases↗
PubMed1989

Chronic hypotensive effects of verapamil in angiotensin hypertension are steroid independent.

This study was designed to examine the mechanisms that contribute to the chronic hypotensive effects of verapamil during angiotensin II-induced hypertension. Hypertension was induced in five dogs by continuous intravenous infusion of angiotensin II (5 ng/kg/min) for 17 days. On the sixth day of angiotensin II infusion when daily sodium balance was achieved, mean arterial pressure (control, 92 +/- 4 mm Hg), plasma aldosterone concentration (control, 5.2 +/- 0.9 ng/dl), and renal resistance (control, 0.28 +/- 0.01 mm Hg/ml/min) were increased 37 +/- 8 mm Hg, 13.6 +/- 5.0 ng/dl, and 0.20 +/- 0.05 mm Hg/ml/min, respectively. At this time there were no significant changes in glomerular filtration rate, effective renal plasma flow, net sodium and water balance, or extracellular fluid volume. Subsequently, when verapamil was infused (at 2 micrograms/kg/min) simultaneously with angiotensin II (days 7-13), there was a net loss of 55 +/- 10 meq sodium, a 7.0 +/- 0.7% fall in extracellular fluid volume, and approximately a 70% reduction in the chronic effects of angiotensin II on mean arterial pressure and renal resistance; in contrast, verapamil failed to attenuate the long-term aldosterone response to angiotensin II. Further, although glomerular filtration rate and effective renal plasma flow tended to increase during verapamil administration, there were no consistent chronic long-term changes in these renal indexes. In comparison with these responses in hypertensive dogs, when verapamil was infused for 7 days before the induction of angiotensin II hypertension, there were no significant changes in any measurements except mean arterial pressure, which fell 11 +/- 1 mm Hg. Thus, these data fail to support the hypothesis that the chronic stimulatory actions of angiotensin II on aldosterone secretion are dependent on a sustained increase in transmembranal calcium influx. Moreover, these data indicate that the pronounced long-term hypotensive effects of verapamil in angiotensin II hypertension are due to impairment of the direct renal actions of angiotensin II rather than the indirect sodium-retaining effects that are mediated via aldosterone secretion.

Aldosterone↗
PubMed1989

Ewing's sarcoma of the foot.

Ewing's sarcoma is a rare, malignant neoplasm that is most common in children and adolescents. It is common in both the axial and appendicular skeleton but has been rarely reported in the foot. Historically, treatment has consisted of chemotherapy and radiation therapy. The roll of surgery in the management of these lesions have recently been "rediscovered" and depends on the anatomic location. The purpose of this paper is to discuss five consecutive cases of Ewing's Sarcoma of the forefoot treated with below-the-knee amputation for local tumor control. Four of the five patients also underwent some systemic chemotherapy. After comparing the various forms of therapy for this tumor, the benefits of below-the-knee amputation make it an excellent primary treatment for Ewing's sarcoma of the foot.

Adolescent↗