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Biomedical subjects

J Walker

Publications and source records attributed to J Walker.

At least 181 records · Page 10Linked to original sources
PubMed1997

Parasite sulphur amino acid metabolism.

This paper reviews current knowledge regarding the metabolism of the sulphur-containing amino acids methionine and cysteine in parasitic protozoa and helminths. Particular emphasis is placed on the unusual aspects of parasite biochemistry which may present targets for rational design of antiparasite drugs. In general, the basic pathways of sulphur amino acid metabolism in most parasites resemble those of their mammalian hosts, since the enzymes involved in (a) the methionine cycle and S-adenosylmethionine metabolism, (b) the trans-sulphuration sequence, (c) the transminative catabolism of methionine, (d) the oxidative catabolism of cysteine and (e) glutathione synthesis have been demonstrated variously in several helminth and protozoan species. Despite these common pathways, there also exist numerous differences between parasite and mammalian metabolism. Some of these differences are relatively subtle. For example, the biochemical properties (and primary amino acid structures) of certain parasite methionine cycle enzymes and S-adenosylmethionine decarboxylases differ from those of the corresponding mammalian enzymes, and nematodes and trichomonads possess a novel, non-mammalian form of the trans-sulphuration enzyme cystathionine beta-synthase. The most profound differences between parasite and mammalian biochemistry relate to a number of unusual enzymes and thiol metabolites found in parasitic protozoa. In certain protozoa the pathway for methionine recycling from 5'-methylthioadenosine differs markedly from the mammalian route, and involves 2 exclusively microbial enzymes. Trypanosomatid protozoa contain the non-mammalian antioxidant thiol compounds ovothiol A and trypanothione, together with unique trypanothione-linked enzymes. Specific anaerobic protozoa possess another exclusively microbial enzyme, methionine gamma-lyase, which catabolises methionine (and homocysteine); the physiological significance of these non-mammalian activities is not fully understood. These unusual features offer opportunities for chemotherapeutic exploitation, and in some cases represent metabolic similarities with bacteria. Additionally, some anaerobic protozoa contain unidentified thiols and this implies the presence of further unusual enzymes/pathways in these organisms. So far, no truly unique targets for chemotherapy have been found in helminth sulphur amino acid metabolism, and to some degree this reflects the relative lack of detailed study in the area.

Amino Acids, Sulfur↗
PubMed1997

Ostertagia circumcincta: isolation of a partial cDNA encoding an unusual member of the mitochondrial processing peptidase subfamily of M16 metallopeptidases.

A reverse-transcriptase polymerase chain reaction (PCR) procedure was used to isolate an Ostertagia circumcincta partial cDNA encoding a protein with general primary sequence features characteristic of members of the mitochondrial processing peptidase (MPP) subfamily of M16 metallopeptidases. The structural relationships of the predicted protein (Oc MPPX) with MPP subfamily proteins from other species (including the model free-living nematode Caenorhabditis elegans) were examined, and Northern analysis confirmed the expression of the Oc mppx gene in adult nematodes.

Amino Acid Sequence↗
PubMed1997

Analysis of acetate counter ion and inorganic impurities in pharmaceutical drug substances by capillary ion electrophoresis with conductivity detection.

Capillary electrophoresis with conductivity detection is a versatile new method for the analysis of counter ions in pharmaceutical drug substances. It is a sensitive and linear technique for determining inorganic ions and short chain carboxylic acids such as acetate. Both acetate counter ion and inorganic impurities can be separated and determined in the same assay.

Acetates↗
PubMed1997

Why are you waiting? Formulating an information pamphlet for use in an accident and emergency department.

The provision of information to patients and relatives reduced their anxiety and leads to greater understanding of what is happening to them and around them. Supplying information verbally has been shown to be at times fragmented and haphazard, particularly when dealing with worried and anxious patients in a hospital setting. The anxiety generated by injury or illness leading to attendance at Accident and Emergency (A & E) Departments does not lend itself to the use of verbal information. To overcome this an information leaflet was developed to supplement verbal information. It describes the working and geography of the A & E department, in an attempt to improve patients' understanding of how the department works and why they may have to wait. The introduction of the leaflet is to supplement verbal communication. Its use has had a positive effect, dovetailing with verbal communication to improve patients' understanding of why they may have to wait.

Emergency Service, Hospital↗
PubMed1997

Expectations of life and health among spinal cord injured adults.

While our understanding of aging and mortality in spinal cord injury is evolving, precise estimates are still not available for expectations of life and health following a spinal cord injury. In order to derive these estimates, information about mortality and health must be combined into a single estimate. Health expectancy estimates have been widely used in the literature of the last decade to try to understand the relationship between population health and survival, both in the general population and in special populations. This study brought the benefit of this methodology to the question of long-term survival following spinal cord injury. Specifically, the study aimed to calculate life and health expectancy in a population of spinal cord injured individuals; and to estimate the effect of factors associated with survival and health. The study involved a retrospective cohort, all of whom sustained a spinal cord injury between the ages of 25 and 34 years, and between 1945 and 1990. The study predicted a median survival time of 38 years post-injury, with 43% surviving at least 40 years. These findings suggest an increase in life expectancy of about 5 years over previous research on the same cohort. Factors affecting survival were age at injury, level and completeness of lesion. Expectations of health found in the present study are similar to those found in studies of the general population. This study showed seven remaining years of poor health expected at injury, and five remaining years expected at 40 years post injury, presumably occurring at the end of life.

Adult↗
PubMed1997

Preterm infants and STRESS: a tool for the neonatal nurse.

Preterm infants are not physiologically or developmentally prepared for life outside the supportive environment of the mother's womb. Their response to stimuli is often immature and disorganized rather than adaptive. The Roy Adaptation Model's theory of an adaptive person may provide a framework for nurses to assess, plan, and evaluate nursing care for fragile preterm infants. The article examines actual and potential stressors of the premature infant; describes commonly observed disorganized, ineffective responses; and proposes a clinical tool (the STRESS tool: signs of stress, touch interventions, reduction of pain, environmental considerations, state, and stability) that nurses can use when caring for medically fragile infants.

Adaptation, Physiological↗
PubMed1997

Synthesis and secretion of von Willebrand factor and fibronectin in megakaryocytes at different phases of maturation.

Our goals have been to define the biochemical characteristics of megakaryocytes during maturation that are critical for platelet assembly and release into the circulation and to introduce biochemical markers for megakaryocytes. To achieve these goals, we have studied fibronectin (FN) and von Willebrand factor (vWF), which are large adhesive proteins that are synthesized by megakaryocytes, stored in alpha granules, and thought to have a fundamental role in hemostasis. The study demonstrated that vWF is primarily synthesized in mature megakaryocytes, which synthesized 7.5 times more vWF than immature megakaryocytes. Brefeldin A, which blocks the exit of proteins from the rough endoplasmic reticulum (RER), inhibited the formation of vWF multimers but did not affect the synthesis of monomers and dimers in mature megakaryocytes. These data are consistent with the formation of vWF dimers in the RER and the assembly of vWF multimers in the trans- and post-golgi. The synthesis of both the 260-kD and 275-kD pro-vWF was detected. However, the synthesis of 275-kD pro-vWF and 220-kD mature vWF was only evident after 2 hours, suggesting that the transit time of nascent vWF through the RER is about 2 hours. Constitutive secretion of vWF was demonstrated in megakaryocytes. About 14.5% and 4.6% of synthesized vWF was secreted by mature and immature megakaryocytes, respectively. In contrast, the synthesis of FN monomers and dimers was established in immature megakaryocytes, and their synthesis in mature megakaryocytes was very similar. Constitutive secretion of FN was not seen in megakaryocytes. Brefeldin A did not inhibit the synthesis of FN dimers; thus, formation of FN dimers occurs in the RER. The demonstration that vWF and FN are synthesized at different phases of megakaryocyte maturation and that only vWF is constitutively secreted by megakaryocytes provides new information relevant to alpha granule formation and possibly bone marrow matrix assembly.

Animals↗
PubMed1997

Labriform propulsion in fishes: kinematics of flapping aquatic flight in the bird wrasse Gomphosus varius (Labridae)

Labriform, or pectoral fin, propulsion is the primary swimming mode for many fishes, even at high relative speeds. Although kinematic data are critical for evaluating hydrodynamic models of propulsion, these data are largely lacking for labriform swimmers, especially for species that employ an exclusively labriform mode across a broad range of speeds. We present data on pectoral fin locomotion in Gomphosus varius (Labridae), a tropical coral reef fish that uses a lift-based mechanism to fly under water at sustained speeds of 1­6 total body lengths s-1 (TL s-1). Lateral- and dorsal-view video images of three fish swimming in a flow tank at 1­4 TL s-1 were recorded at 60 Hz. From the two views, we reconstructed the three-dimensional motion of the center of mass, the fin tip and two fin chords for multiple fin beats of each fish at each of four speeds. In G. varius, the fin oscillates largely up and down: the stroke plane is tilted by approximately 20 ° from the vertical. Both frequency and the area swept by the pectoral fins increase with swimming speed. Interestingly, there are individual differences in how this area increases. Relative to the fish, the fin tip in lateral view moves along the path of a thin, inclined figure-of-eight. Relative to a stationary observer, the fin tip traces a sawtooth pattern, but the teeth are recumbent (indicating net backwards movement) only at the slowest speeds. Distal fin chords pitch nose downward during the downstroke and nose upward during the upstroke. Hydrodynamic angles of attack are largely positive during the downstroke and negative during the upstroke. The geometry of the fin and incident flow suggests that the fin is generating lift with large upward and small forward components during the downstroke. The negative incident angles during the upstroke suggest that the fin is generating largely thrust during the upstroke. In general, the large thrust is combined with a downward force during the upstroke, but the net backwards motion of the fin at slow speeds generates a small upward component during slow swimming. Both the alternating sign of the hydrodynamic angle of attack and the observed reduced frequencies suggest that unsteady effects are important in G. varius aquatic flight, especially at low speeds. This study provides a framework for the comparison of aquatic flight by fishes with aerial flight by birds, bats and insects.

Journal Article↗
PubMed1997

Motor patterns of labriform locomotion: kinematic and electromyographic analysis of pectoral fin swimming in the labrid fish Gomphosus varius

Labriform locomotion is a widespread swimming mechanism in fishes during which propulsive forces are generated by oscillating the pectoral fins. We examined the activity of the six major muscles that power the pectoral fin of the bird wrasse Gomphosus varius (Labridae: Perciformes). The muscles studied included the fin abductors (arrector ventralis, abductor superficialis and abductor profundus) and the fin adductors (arrector dorsalis, adductor superficialis and adductor profundus). Our goals were to determine the pattern of muscle activity that drives the fins in abduction and adduction cycles during pectoral fin locomotion, to examine changes in the timing and amplitude of electromyographic (EMG) patterns with increases in swimming speed and to correlate EMG patterns with the kinematics of pectoral fin propulsion. EMG data were recorded from three individuals over a range of swimming speeds from 15 to 70 cm s-1 (1­4.8 TL s-1, where TL is total body length). The basic motor pattern of pectoral propulsion is alternating activity of the antagonist abductor and adductor groups. The downstroke is characterized by activity of the arrector ventralis muscle before the other abductors, whereas the upstroke involves nearly synchronous activity of the three adductors. Most EMG variables (duration, onset time, amplitude and integrated area) showed significant correlations with swimming speeds. However, the timing and duration of muscle activity are relatively constant across speeds when expressed as a fraction of the stride period, which decreases with increased velocity. Synchronous recordings of kinematic data (maximal abduction and adduction) with EMG data revealed that activity in the abductors began after maximal adduction and that activity in the adductors began nearly synchronously with maximal abduction. Thus, the pectoral fin mechanism of G. varius is activated by positive work from both abductor and adductor muscle groups over most of the range of swimming speeds. The adductors produce some negative work only at the highest swimming velocities. We combine information from pectoral fin morphology, swimming kinematics and motor patterns to interpret the musculoskeletal mechanism of pectoral propulsion in labrid fishes.

Journal Article↗
PubMed1997

C5b-7 and C5b-8 precursors of the membrane attack complex (C5b-9) are effective killers of E. coli J5 during serum incubation.

The finding that C9-deficient sera (C9D) can kill serum sensitive strains of Gram-negative bacteria by us and other investigators, questions the role of C9 in the membrane attack complex as necessary for cell death. In these studies we have demonstrated that C5b-8 complexes generated on E. coli J5 during incubation in C9-depleted and C9-neutralized sera are effective in killing Gram-negative bacteria. In the same study, we extended our investigations to show that the deposition of C5b-7 complexes (from C8-deficient [C8D], C8 depleted and C8-neutralized sera) is also effective in killing Gram-negative bacteria. In all cases, these studies demonstrated that when E. coli J5 was incubated with C8D, C9D and pooled normal human serum [PNHS], deposited C5b-9 complexes from PNHS produced more killing than C5b-7 or C5b-8 complexes alone. These experiments clearly demonstrated that C5b-7 and C5b-8 complexes are bactericidal and that multimeric C9 within C5b-9 is not an absolute requirement for inner membrane damage and cell death of Gram-negative bacteria.

Blood Bactericidal Activity↗
PubMed1997

Characterization of cytokine profiles and double-positive lymphocyte subpopulations in normal bovine lungs.

OBJECTIVE: To characterize cytokine profiles and lymphocyte subpopulations in lung parenchyma and bronchoalveolar lavage (BAL) fluid from normal bovine lungs. ANIMALS: Eight 12- to 18-month-old cattle. PROCEDURE: Cell populations in BAL fluid and collagenase-digested lung parenchyma were analyzed by flow cytometry and monoclonal antibodies. Proportions of total cell populations were determined, using Giemsa-stained cytospots. Distribution of lymphocytes within the lung parenchyma was analyzed by immunohistochemistry, and cytokine mRNA species in the parenchyma were characterized by use of reverse transcriptase-polymerase chain reaction analysis. RESULTS: Cytokine profiles indicated high amounts of mRNA for interleukins 6 and 10 and transforming growth factor beta. In the BAL fluid and lung parenchyma, macrophages were the predominant cell type, although the proportion was lower in the parenchyma. Lymphocytes made up approximately 3% of both cell populations. Common to both lung compartments was the predominance of CD2+ and gamma delta T cells over B lymphocytes. There were more CD8+ T cells than CD4+ T cells in both compartments. The gamma delta cells made up approximately 9% of the lymphocyte populations. Two-color flow cytometry revealed CD8+ gamma delta T cell and CD8+CD5- populations that were unique to BAL fluid. In the BAL fluid and parenchyma, most CD4+ and CD8+ T cells expressed high amounts of CD44, a characteristic of memory T cells. The gamma delta T cells were CD44(10), as were B cells in the lung parenchyma. The B cells from BAL fluid expressed high amounts of CD44. Immunohistologic analysis of lung tissue revealed bronchus-associated lymphoid tissue structures with distinctive germinal center organization of B cells encompassed by CD4+ T cells. CONCLUSIONS: Results provided normal values for comparison with those of other species and with the bovine respiratory tract response to disease.

Animals↗
PubMed1996

Structure of the amino-terminal core domain of the HIV-1 capsid protein.

The three-dimensional structure of the amino-terminal core domain (residues 1 through 151) of the human immunodeficiency virus-type 1 (HIV-1) capsid protein has been solved by multidimensional heteronuclear magnetic resonance spectroscopy. The structure is unlike those of previously characterized viral coat proteins and is composed of seven alpha helices, two beta hairpins, and an exposed partially ordered loop. The domain is shaped like an arrowhead, with the beta hairpins and loop exposed at the trailing edge and the carboxyl-terminal helix projecting from the tip. The proline residue Pro1 forms a salt bridge with a conserved, buried aspartate residue (Asp51), which suggests that the amino terminus of the protein rearranges upon proteolytic maturation. The binding site for cyclophilin A, a cellular rotamase that is packaged into the HIV-1 virion, is located on the exposed loop and encompasses the essential proline residue Pro90. In the free monomeric domain, Pro90 adopts kinetically trapped cis and trans conformations, raising the possibility that cyclophilin A catalyzes interconversion of the cis- and trans-Pro90 loop structures.

Amino Acid Isomerases↗
PubMed1996

Purification of hydrophobic integral membrane proteins from Mycoplasma hyopneumoniae by reversed-phase high-performance liquid chromatography.

A general and practical approach for isolating, fractionating and purifying large quantities of outer membrane hydrophobic proteins is described as applied to membrane proteins of Mycoplasma hyopneumoniae. Outer membrane proteins were extracted with Triton X-114 detergent and were precipitated from the detergent phase with 90% ethanol. Precipitated proteins were dissolved in 65% formic acid and separated by RP-HPLC using a formic acid-acetonitrile gradient. A M(r) 48 000 protein was obtained in high yield and at greater than 90% purity by optimisation of parameters for RP-HPLC. The combination of Triton X-114 extraction followed by high resolution RP-HPLC is a novel and rapid procedure for the isolation and purification of hydrophobic proteins. Proteins purified by this approach were suitable for subsequent characterisation by direct sequencing of the amino terminus as well as generation of peptides by digestion with cyanogen bromide.

Amino Acid Sequence↗
PubMed1996

Travel of U.S. Citizens after Coronary Artery Bypass Surgery.

Background: International travel with concurrent medical problems is commonplace. This study was undertaken to determine travel morbidity rates in coronary artery bypass surgery (CABS) patients. Methods: A survey of 10,096 CABS patients indicated that 40% (n=4286) of them reported travel during the study year of 1988. The patients who reported traveling and those patients who did not travel were stratified by gender and by age (30 to 54, 55 to 64, and 65 to 80 years). Results: The male patients who traveled reported a 6.1% incidence of chest pain during travel. The comparable female patients reported an 8.7% incidence of chest pain (p <.05). Hospitalization and myocardial infarction rates of patients during travel were low (0.6% and 0.2%, respectively). In the older age groups (55 to 64 years and 65 to 80 years), males who traveled were less likely to report angina than the male patients who did not travel (p <.05). Conclusions: A history of CABS and the stress of travel do not appear to be associated with "high" hospitalization rates or special medical problems.

Journal Article↗
PubMed1996

The acylation of megakaryocyte proteins: glycoprotein IX is primarily myristoylated while glycoprotein Ib is palmitoylated.

The acylation of megakaryocyte proteins was studied with special emphasis on the myristoylation and palmitoylation of the glycoprotein (GP) Ib complex. Guinea pig megakaryocytes were purified and separated into subpopulations at different phases of maturation. Cells were incubated with [3H]myristate, [3H]palmitate, or [3H]acetate to study endogenous protein acylation. Cycloheximide was used to distinguish between cotranslational and posttranslational acylation and hydroxylamine to distinguish between thioester and amide linkages. After incubations, delipidated proteins or GPIb complex subunits, immunoprecipitated with PG-1, AN-51 or FMC-25 monoclonal antibody, were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and assessed by fluorography. Radiolabeled fatty acids bound to GPIX and GPIb were also analyzed by high pressure liquid chromatography (HPLC) and scintillation spectrometry. With [3H]myristic acid and [3H]acetate, GPIX was found to be a major myristoylated protein in megakaryocytes and CHRF-288 cells. Myristic acid was linked to GPIX by an amide bond, and this process occurred cotranslationally. With [3H]acetate, GPIb was primarily palmitoylated, but with [3H]myristate, GPIb was acylated with about equal mounts of myristic acid and palmitic acids. Both fatty acids were linked to GPIb by thioester bonds, and acylation was posttranslational. The myristoylation of GPIX while the palmitoylation of GPIb occurred throughout megakaryocyte maturation. Myristoylation and palmitoylation may have different functions relevant to the assembly of the GPIb complex in megakaryocytes.

Acylation↗
PubMed1996

Unmasking mRNA in clam oocytes: role of phosphorylation of a 3' UTR masking element-binding protein at fertilization.

During meiotic maturation or after fertilization of invertebrate and vertebrate oocytes, many of the quiescent stored mRNAs are recruited into polysomes. In the clam, Spisula solidissima, such masked messages include the abundant mRNAs encoding cyclin A and the small subunit of ribonucleotide reductase. We have previously shown that mRNA-specific unmasking of these two messages can be achieved in vitro, in oocyte cell-free extracts, by the addition of antisense RNAs corresponding to a fairly short (130-140 nucleotides) segment in their cognate 3' untranslated regions. We postulated that the antisense RNAs prevented the binding of a masking repressor protein (Standart et al., 1990). Here we report UV-crosslinking and gel retardation studies which show that the masking portions of the translationally regulated mRNAs bind an oocyte protein of 82 kDa (p82), which is phosphorylated after fertilization. This modification was accompanied by altered RNP complex formation in gel retardation assays. These changes presumably reflect the activation of translation of the masked mRNAs. The role of p82 phosphorylation in maternal mRNA unmasking was assessed in a novel in vitro activation system developed from clam oocytes, based upon the natural rise in pH which accompanies fertilization. Concomitant with mRNA unmasking, several kinases, including cdc2 and MAP kinases were activated in this system, as was p82 phosphorylation. Inhibitors of serine/threonine kinases, including 6-DMAP, staurosporine, and H7 inhibited p82 phosphorylation, whereas inhibitors of tyrosine kinases, protein kinase C, cAMP-dependent protein kinase, and p70s6k did not prevent this modification. A specific inhibitor of cdc2 kinase, p27Kip1, prevented p82 phosphorylation and translational activation, strongly suggesting that p82 modification is required for unmasking.

Amino Acid Sequence↗