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Biomedical subjects

R A Walters

Publications and source records attributed to R A Walters.

At least 19 recordsLinked to original sources
PubMed1996

Small, low-cost implantable centrifugal pump for short-term circulatory assistance.

BACKGROUND: In 1991, Allegheny General Hospital and Allegheny-Singer Research Institute purchased a centrifugal pump, then a 2-year-old technology, from Medtronic Bio-Medicus, as part of its research program for novel treatments of acute and chronic heart failure. During a 4-year development program, we then established and met goals of durability, performance, thromboresistance, and low cost. METHODS: In vitro testing involved extensive hydraulic characterizations using Penn State mock loops. Calorimetry was used to determine efficiency. Durability studies used heated (37 degrees C) seawater for 28 to 45 days. In vivo studies used 46 sheep to test performance and engineering changes and to determine myocardial oxygen consumption, thromboresistance, and long-term durability. A left atrium-to-aorta circuit was used in all. RESULTS: Hydraulic testing showed no preload sensitivity but moderate afterload sensitivity at all impeller speeds (2,000 to 6,000 rpm). The heat load was low, and overall efficiency was 13% to 15%. Bench durability studies showed no electrical malfunction of the stator or console without degradation of the biomaterials used. Acute in vitro studies showed a near-linear relationship of myocardial oxygen consumption and left ventricular stroke work, pump flow, and pump speed. At speeds of 2 to 3 L/min (50% bypass), left ventricular stroke work and myocardial oxygen consumption were decreased approximately 50%. Additionally, 5 animals have had implants for 28 to 154 days with no macroemboli or microemboli detected in any animal. Hematologic and biochemical studies became normal 3 to 7 days after implantation. Hemolysis was low at less than 10 mg/dL. Clinical costs of the device are estimated to be 80% less than those of currently available devices. CONCLUSIONS: We conclude that an old technology has been made into new technology by application of sound engineering design principles, microchips, and new biomaterials. Qualifying trails for a Food and Drug Agency investigational device exemption application are in progress.

Animals↗
PubMed1990

A practical irreversible hydrocolloid impression tray holder.

A small, practical, inexpensive, and esthetic tray-holding device can be made easily for use in the operatory and laboratory. The device, when not in use, can be conveniently stored in a drawer or on a small shelf and is inconspicuous. Several of these devices can be made at one time and strategically located within the office.

Alginates↗
PubMed1990

An effect of tray design and material retention on the linear dimensional changes in polysulfide impressions.

One-unit fixed partial dentures, which fit accurately on the laboratory dies made from polysulfide impression materials, often pose a problem in seating completely in the mouth of the patient. Sectioning of the fixed partial denture units allows the individual units to seat accurately in the mouth but requires a soldering procedure. Linear distortion of the mercaptan (polysulfide) rubber base that takes place during setting is a cause of this problem. The distortion is directly related to (1) tray design, (2) area of adhesive placement, and (3) excessive bulk of material on the tray border during impression making. A controlled amount of adhesive and impression material in a new design for a custom tray must be used to minimize this distortion. When these factors are controlled, the resulting impression is unrestricted, and a more accurate working die is obtained. This results in a better fit of one-unit fixed partial dentures in the mouth.

Adhesives↗
PubMed1985

Coordinate amplification of metallothionein I and II genes in cadmium-resistant Chinese hamster cells: implications for mechanisms regulating metallothionein gene expression.

We describe here the derivation, characterization, and use of clonal cadmium-resistant (Cdr) strains of the Chinese hamster cell line CHO which differ in their metallothionein (MT) induction capacity. By nondenaturing polyacrylamide gel electrophoresis, we showed that the stable Cdr phenotype is correlated with the augmented expression of both isometallothioneins (MTI and MTII). In cells resistant to concentrations of CdCl2 exceeding 20 microM, coordinate amplification of genes encoding both isometallothioneins was demonstrated by using cDNA MT-coding sequence probes and probes specific for 3'-noncoding regions of Chinese hamster MTI and MTII genes. Molecular and in situ hybridization analyses supported close linkage of Chinese hamster MTI and MTII genes, which we have mapped previously to Chinese hamster chromosome 3. This suggests the existence of a functionally related MT gene cluster in this species. Amplified Cdr variants expressing abundant MT and their corresponding Cds parental CHO cells should be useful for future studies directed toward elucidating the mechanisms that regulate expression of the isometallothioneins.

Animals↗
PubMed1984

Construction and analysis of DNA sequence libraries from flow-sorted chromosomes: practical and theoretical considerations.

We describe the construction and analysis of recombinant DNA libraries representative of chromosomes 1 and 2 of Chinese hamster (Cricetulus griseus). Propidium-iodide stained chromosomes were purified by flow cytometric analysis and sorting, and EcoRI digests of purified DNA were cloned into the bacteriophage vector Charon 4A. These libraries contain DNA complementary to 63% and 69% of nick-translated DNA derived from flow-purified chromosomes 1 and 2, respectively. However, sequences complementary to only 24% and 35% of a total Chinese hamster genomic DNA tracer were hybridized in parallel renaturation experiments. The chromosome 2 library contained DNA sequences encoding dihydrofolate reductase (dhfr), a gene previously mapped to Chinese hamster chromosome 2. No sequences complementary to dhfr were found in the library constructed from chromosome 1 DNA. These analyses are discussed with regard to the current limitations and future strategies for the construction of chromosome-specific DNA sequence libraries of high purity and completeness.

Animals↗
PubMed1983

cDNA cloning and nucleotide sequence comparison of Chinese hamster metallothionein I and II mRNAs.

Polyadenylated RNA was extracted from a cadmium resistant Chinese hamster (CHO) cell line, enriched for metal-induced, abundant RNA sequences and cloned as double-stranded cDNA in the plasmid pBR322. Two cDNA clones, pCHMT1 and pCHMT2, encoding two Chinese hamster isometallothioneins were identified, and the nucleotide sequence of each insert was determined. The two Chinese hamster metallothioneins show nucleotide sequence homologies of 80% in the protein coding region and approximately 35% in both the 5' and 3' untranslated regions. Interestingly, an 8 nucleotide sequence (TGTAAATA) has been conserved in sequence and position in the 3' untranslated regions of each metallothionein mRNA sequenced thus far. Estimated nucleotide substitution rates derived from interspecies comparisons were used to calculate a metallothionein gene duplication time of 45 to 120 million years ago.

Amino Acid Sequence↗
PubMed1981

Differential induction by cadmium of a low-complexity ribonucleic acid class in cadmium-resistant and cadmium-sensitive mammalian cells.

The Chinese hamster ovary (CHO) cell line and the subline Cdr20F4 have been used to compare cadmium-induced ribonucleic acid (RNA) synthesis in cadmium-sensitive and cadmium-resistant cells, respectively. Gel electrophoresis of the cell-free translation products directed by polyadenylated [poly(A+)] messenger RNA (mRNA) from cadmium-induced Cdr20F4 cells revealed four low molecular weight species (Mr 7000-21 000), including metallothionein, whose synthesis was not detected after translation of either cadmium-induced or uninduced CHO cell poly(A+) mRNA. At least two of these species were also detected after translation of an abundant 400-nucleotide (NT) RNA class purified from the cadmium-induced Cdr20F4 cell RNA. Molecular hybridization of complementary deoxyribonucleic acid (cDNA) complementary to this abundant, cadmium-induced 400-NT RNA fraction indicates that the cadmium-induced RNA class possesses a total kinetic complexity of about 2000 NT's. At least half of these inducible sequences are also represented constitutively in less abundant RNA classes of both uninduced CHO and Cdr20F4 cells. Induction of Cdr20F4 cells with cadmium increases the cellular concentration of the 2000-NT-complexity RNA class to a level at least 2 x 10(3)-fold greater than its constitutive level in uninduced Cdr20F4 cells. Induction of CHO cells with cadmium increases the cellular concentration of a subset of the sequences in the 2000-NT-complexity class, but only to a level 100-fold over the constitutive level in uninduced CHO cells. The remainder of these sequences belongs to the least abundant CHO cell poly(A+) RNA class.

Animals↗
PubMed1980

Amino acid analysis and cell cycle dependent phosphorylation of an H1-like, butyrate-enhanced protein (BEP; H1(0); IP25) from Chinese hamster cells.

A fraction enriched in the butyrate-enhanced protein (BEP) has been isolated from Chinese hamster (line CHO) cells by perchloric acid extraction and Bio-Rex 70 chromatography. Amino acid analyses indicate that the composition of BEP resembles that of CHO H1; however, BEP contains 11% less alanine than H1, and, in contrast to H1, BEP contains methionine. Treatment of BEP with cyanogen bromide results in the cleavage of a small fragment of approximately 20 amino acids so that the large fragment seen in sodium dodecyl sulfate--acrylamide gels has a molecular weight of approximately 20 000. Radiolabeling and electrophoresis indicate that BEP is phosphorylated in a cell cycle dependent fashion. In G1-arrested cells, little or no phosphate is incorporated into BEP. As cells progress through interphase, BEP becomes phosphorylated so that 12--35% of the BEP molecules are phosphorylated at one to two sites by late interphase. During mitosis, all BEP molecules become phosphorylated at approximately four sites per molecule (BEPM). Electrophoresis and the analysis of cell populations by electron microscopy indicate that the appearance of BEPM is temporally correlated with the mitotic phosphorylation of histone H1 (H1M) and with chromosomal condensation during prophase, metaphase, and anaphase. During exit from mitosis, BEPM undergoes dephosphorylation. The dephosphorylation of BEPM is temporally correlated with dephosphorylation of H1M and with the unraveling of fully condensed chromosomes near the anaphase--telophase transition. These data suggest that (1) BEP is a specialized histone of the H1 class and (2) BEP is the species equivalent of calf lung histone H1(0) [Panyim, S., & Chalkley, R. (1969) Biochem. Biophys. Res. Commun. 37, 1042], rat H1(0) [Medvedev, Zh. A., Medvedeva, M. N., & Huschtscha, L. I. (1977) Gerontology (Basel) 23, 334], and IP25, a protein enhanced in differentiated Friend erythroleukemia cells [Keppel, F., Allet, B., & Eisen, H. (1977) Proc. Natl. Acad. Sci. U.S.A. 74, 653]. The data also indicate that putative HMG1 and HMG2 proteins do not undergo the extensive cell cycle dependent phosphorylations measured for histone H1 and BEP.

Amino Acids↗
PubMed1979

Comparison of complexity and diversity of polyadenylated polysomal and informosomal messenger ribonucleic acid from Chinese hamster cells.

The sequence complexity and relative abundance of cytoplasmic polyadenylated polysomal (ribosome-bound) mRNA and cytoplasmic polyadenylated informosomal (ribosome-free) mRNA were analyzed in exponentially growing Chinese hamster cells (line CHO) using the technique of cDNA hybridization to excess poly(A)+ mRNA. Polysomal and informosomal mRNAs had similar complexities ( approximately 8300 mRNA species), but both the fraction of mRNA and the number of sequences comprising the mRNA abundance classes were different. Heterologous annealing reactions showed that all of the mRNA sequences detected were shared by the polysomal and informosomal mRNAs. However, the most abundant informosomal mRNA component was considerably different from the most abundant polysomal mRNA component. For a more detailed analysis, cDNA complementary to the most abundant informosomal and polysomal mRNAs was isolated. By use of the fractionated cDNA, it could be demonstrated that the most abundant informosomal mRNA sequences were distributed in the polysomal mRNA with an approximately fivefold reduction in relative frequency. These results are not compatible with models postulating translational control of gene expression by the complete sequestering of some mRNA sequences in an untranslatable form in the cytoplasm. The data are, however, consistent with models encompassing differential rates of initiation on the polysome and/or preferential affinity of some mRNAs for initiation factors.

Animals↗
PubMed1978

Action of heparin on mammalian nuclei. II. Cell-cycle-specific changes in chromatin organization correlate temporally with histone H1 phosphorylation.

The interaction of the polyanion heparin with the inner histones of chromatin has been used to detect changes in chromatin organization associated with cell-cycle traverse. Synchronized populations of Chinese hamster cells were obtained either in early G1 or near the G1/S boundary. The rate of interaction of heparin with chromatin-associated inner histones was measured using nuclei isolated from synchronized cell populations in different phases of the cell cycle. A G1-specific decrease in rate of interaction of heparin with inner histones was observed and found to be independent of the presence of hydroxyurea during traverse of G1. A further decrease in heparin-inner histone interaction occurred in late S and G2. These changes correlate temporally with the interphase phosphorylation(s) of histone H1. This correlation is discussed within the framework of current models of higher order chromatin structure (i.e. organization above the nucleosome level). Analysis of the cooperativity of interaction of heparin with inner histones was performed using the kinetic analog of the Hill equation. This analysis suggests that the organization of inner histones on chromatin does not undergo large variations during the cell cycle.

Cell Cycle↗