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Biomedical subjects

R Cook

Publications and source records attributed to R Cook.

At least 37 records · Page 2Linked to original sources
PubMed1989

Interaction of tetrahydropteroylpolyglutamates with two enzymes from mitochondria.

The dissociation constants of tetrahydropteroylpolyglutamates, having from one to six glutamate residues, have been determined for the two mitochondrial enzymes serine hydroxymethyltransferase and dimethylglycine dehydrogenase. The ratios of the dissociation constants for the mono- and hexaglutamate forms of the coenzyme were 200 and less than 10 for serine hydroxymethyltransferase and dimethylglycine dehydrogenase, respectively. Km and kcat values were determined for the reversible interconversion of serine and glycine as a function of the number of glutamyl residues on the coenzyme. The values in the serine to glycine direction did not significantly change with the number of glutamyl residues, but in the glycine to serine direction, there was a 9-fold increase in the kcat/Km when the longer chain polyglutamates were used as the coenzyme substrate. A sensitive and rapid method for determining the dissociation constants of proteins which bind either tetrahydropteroylpolyglutamates or their 5-methyl and 5-formyl conjugates is described.

Animals↗
PubMed1989

The role of confidential enquiries in the reduction of maternal mortality and alternatives to this approach.

The aim of confidential enquiries into maternal deaths is to identify weaknesses in the maternal health care system with a view to remedying them. The method of confidential enquiry is explained using the British system as an example. The reasons why this apparently useful practice is not more widely adopted can in some countries include fears of litigation or lack of trust in confidentiality. Alternative approaches to maternal death audit are discussed.

Cause of Death↗
PubMed1989

Identification of consensus epitope structures expressed in recombinant DNA libraries.

In the course of screening cDNA expression libraries with a monospecific polyclonal antibody to topoisomerase I, we isolated three different immunopositive cDNA clones. By comparing their derived amino acid sequences, a consensus region of similarity in otherwise completely dissimilar sequences was identified as an epitope. The approach described here should identify both continuous and discontinuous topological epitopes. The probability of occurrence of "spurious" immunopositive clones is shown to depend on the number of codons of each critical amino acid within the antigenic determinant.

Amino Acid Sequence↗
PubMed1989

Recombinant tissue-type plasminogen activator in canine embolic pulmonary hypertension. Effects of bolus versus short-term administration on dynamics of thrombolysis and on pulmonary vascular pressure-flow characteristics.

We used a canine model of embolic pulmonary hypertension, induced by injection of autologous radioactive blood clots, to investigate effects of recombinant tissue-type plasminogen activator (rt-PA) on dynamics of thrombolysis and on pulmonary pressure-flow (PQ) characteristics. Over 5 (rt-PA5) or 15 (rt-PA15) minutes, 1 mg/kg rt-PA was infused. Rate and extent of thrombolysis were assessed by counting over both lung fields with a gamma camera. Emboli increased mean pulmonary artery pressure from 14 to 36 mm Hg (p less than 0.005). This change was predominantly due to an increase in the effective outflow pressure (PI) (from 9 to 29 mm Hg, p less than 0.001), obtained by extrapolation from the linear PQ relation. While pulmonary hemodynamics improved with rt-PA5 and rt-PA15, the change was greatest with rt-PA15. For example, the increase in PI that occurred with embolization was abolished with rt-PA15. Also, the decrease in pulmonary artery pressure was greatest with rt-PA15. While not significantly different, extent of total clot lysis tended to be greatest with rt-PA15 (p less than 0.07). Also, while during infusion, the concentration of rt-PA5 was threefold that of rt-PA15, the corresponding rate of thrombolysis was similar with rt-PA5 and rt-PA15. These results indicate that the improvement in pulmonary hemodynamics with rt-PA is primarily explained by a decrease in PI. Furthermore, they suggest an upper limit to the dose-thrombolytic rate relation with rt-PA.

Animals↗
PubMed1988

Mutations affecting assembly of beta-tubulin localize to a region near the carboxyl terminus.

The generation of Chinese hamster ovary cell lines that express assembly defective forms of beta-tubulin were isolated using selections based on reversion of conditional lethal or drug resistance phenotypes. Two such cell lines, D2 and 6H3, were chosen for further characterization because they contain beta-tubulin polypeptides that exhibit decreases in apparent molecular weight on two-dimensional gel electrophoresis. Analysis of the nucleic acid from these cell lines using both Southern and Northern procedures suggests a deletion in one of the beta-tubulin genes in each cell line. Localization of the missing sequence in D2 was first determined by tryptic peptide mapping by high performance liquid chromatography. Subsequently, the assignment was confirmed by constructing appropriate subclones of a wild type Chinese hamster ovary beta-tubulin cDNA for Southern analysis to demonstrate a failure to recognize characteristic hybridization patterns of the mutant tubulin gene. In the other revertant, 6H3, the deletion was detected on a Northern blot by differential hybridization of a 3' fragment of the cDNA to the beta-tubulin messages. The results indicate that D2 has an internal deletion whose approximate limits extend from amino acid residues 250 through 345. Cell line 6H3 has a deletion that begins near amino acid residue 330 and extends into the 3'-untranslated region of the gene.

Animals↗
PubMed1988

Partial amino acid sequence of rat topoisomerase I: comparison with the predicted sequences for the human and yeast enzymes.

The partial amino acid sequence of rat topoisomerase I was determined by gas-phase microsequencing. Seven tryptic peptides closely matched the sequences deduced from human topoisomerase I cDNA (94.5% homology). Similarity to sequences deduced from baker's yeast and fission yeast genomic DNA were restricted to conserved domains which may represent important sites of interaction with DNA or with other proteins.

Amino Acid Sequence↗
PubMed1988

Sequences of the VH and VL regions of murine monoclonal antibodies against 3-fucosyllactosamine.

Many mAb that bind the carbohydrate antigenic determinant 3-fucosyl-lactosamine (3-FL), Gal beta 1-4[Fuc alpha-3]GlcNAc-R have been raised in BALB/c mice, and we are studying the structure and regulation of these antibodies. In this report, we present the first information about their amino acid sequences and the Ig gene segments used to encode them. V regions of the H and L chains of three anti-3-FL antibodies, PMN6, PMN29, and PM81, were sequenced by a combination of mRNA and amino acid sequencing. The L chain sequences of PMN6 and PM81 antibodies indicate that their VK and JK regions are encoded by VK24B and JK1 germ-line genes, respectively. The nucleotide and amino acid sequences of the H chains suggest that the three anti-3-FL antibodies are encoded by the VH441 gene segment of the X24 VH family, and this conclusion was supported by Southern filter hybridization with VH441 and JH3-JH4 probes. PMN29 has at least 11 amino acid substitutions, which is an unusually large amount of somatic mutation for an IgM antibody. Previous analyses of BALB/c genomic libraries with VHX24 and VH441 probes make it unlikely that this VH family contains additional germ-line genes, but this possibility cannot be excluded. All three antibodies use the DQ52 and JH4 gene segments. The single VH and VL gene segments used to encode the anti-3-FL antibodies is in contrast to the multiple VH and VL segments used by antibodies against other carbohydrate Ag such as alpha 1-6 dextran and group A streptococcal carbohydrate. VH441 also encodes the VH regions of antibodies against galactan and levan (beta 2-6 fructosan). The similarities among VH segments of antibodies against 3-FL, levan, and galactan, and the striking differences in their CDR3 sequences, suggest that CDR3 plays an important role in the formation of the Ag binding site. The use of a single VH segment from the smallest VH gene family by antibodies against at least three different carbohydrate determinants is noteworthy. It raises the possibility that the amino acid sequence encoded by VH441 has some general structural features that make it particularly well adapted for binding to carbohydrate sequences.

Amino Acid Sequence↗
PubMed1988

Treatment of canine embolic pulmonary hypertension with recombinant tissue plasminogen activator. Efficacy of dosing regimes.

We investigated effects of two dosing regimes of recombinant tissue plasminogen activator (rt-PA) and sodium heparin on pulmonary thrombolysis in a canine model of pulmonary hypertension, induced by injection of radioactive blood clots. By continuously counting over both lung fields with a mobile gamma camera, we correlated rate and extent of pulmonary thrombolysis with corresponding pulmonary hemodynamics. Treatment with heparin, over a 3-hour interval, did not result in significant thrombolysis or in a decrease in mean pulmonary artery pressure (PAP). In contrast, rt-PA caused marked pulmonary thrombolysis. While total clot lysis was similar when 1 mg/kg rt-PA was infused over 15 (rt-PA15) or 90 (rt-PA90) minutes (47% and 42%, respectively), rate of lysis during infusion was markedly increased with rt-PA15 (56% vs. 27%/hr, p less than 0.001). Corresponding to the increased rate of thrombolysis with rt-PA15, relative PAP decrease was greater at 15 and 30 minutes. At 4 hours, PAP decreased most with rt-PA90. However, two of the six dogs given rt-PA15 had an increase in PAP and lung radioactivity 1 hour after rt-PA. This was associated with dislodgment of a previously trapped clot. These results suggest that rt-PA may be appropriate therapy for pulmonary embolism and support further studies designed to optimize dosing regimes.

Animals↗
PubMed1988

A non-flow cytometric system for detecting antibodies and cellular antigens in blood.

The system described here can distinguish between single and agglutinated erythrocytes by use of a non-flow fiber optic fluorometer. The method is capable of detecting cell-surface antigens and antibodies to cell-surface antigens present in blood. Significant features include high-efficiency fluorescent dyes that intercalate into cell membranes, a stretching membrane for transport and mixing of samples, charged colloidal magnetite for magnetic separation of erythrocytes, and an immersible fiber optic probe for measuring fluorescence associated with cells in a 1-nL volume of a bulk solution. We describe the application of the system to automation of ABO/Rh grouping and antibody screening.

ABO Blood-Group System↗
PubMed1985

The alternatives approach revisited: a biopsychological model and guidelines for application.

The alternatives approach to drug abuse prevention is based on the concept that individuals provided with healthful, nonchemical ways of gaining rewards and pleasures will be less likely to engage in drug or alcohol abuse. After enjoying a brief period of popularity the approach seems to have fallen out of favor, largely because of a combination of conceptual confusion and the lack of any systematic guidelines for the development and presentation of alternative activities. This article seeks to remedy those twin deficiencies. An attempt is made to explicate the physical and psychological dynamics underlying alternatives, culminating in the formulation of a biopsychological model of alternatives. The model depicts the sequence of events which moves an individual toward or away from healthful activities or drug use, and it generates a series of predictions and potential empirical tests of the model. The article presents a series of practical guidelines for the selection and development of alternative activities, designed to assist and encourage the development of healthful activities in the schools, the home, and the community.

Adaptation, Psychological↗
PubMed1979

Sensory extinction and sensory reinforcement principles for programming multiple adaptive behavior change.

The role of sensory reinforcement was examined in programming multiple treatment gains in self-stimulation and spontaneous play for developmentally disabled children. Two phases were planned. First, we attempted to identify reinforcers maintaining self-stimulation. Sensory Extinction procedures were implemented in which auditory, proprioceptive, or visual sensory consequences of self-stimulatory behavior were systematically removed and reintroduced in a reversal design. When self-stimulation was decreased or eliminated as a result of removing one of these sensory consequences, the functional sensory consequence was designated as a child's preferred sensory reinforcer. In Phase 2, we assessed whether children would play selectively with toys producing the preferred kind of sensory stimulation. The results showed the following. (1) Self-stimulatory behavior was found to be maintained by sensory reinforcement. When the sensory reinforcer was removed, self-stimulation extinguished. (2) The sensory reinforcers identified for self-stimulatory behavior also served as reinforcers for new, appropriate toy play. (3) The multiple treatment gains observed appeared to be relatively durable in the absence of external reinforcers for play or restraints on self-stimulation. These results illustrate one instance in which multiple behavior change may be programmed in a predictable, lawful fashion by using "natural communities of sensory reinforcement."

Autistic Disorder↗