Erythropoietin escape in five CAPD patients.
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Adult↗
Biomedical subjects
P Williams
Publications and source records attributed to P Williams.
Evidence for the presence of a proteinase-activated receptor distinct from the thrombin receptor in vascular endothelial cells.
The thrombin receptor was the first cloned G protein-coupled receptor reported to be activated by proteolytic cleavage of its extracellular amino terminus. A second proteinase-activated receptor (PAR-2) was cloned recently and expressed in Xenopus laevis oocytes. PAR-2 was activated by trypsin and by a peptide (SLIGRL) derived from the new amino terminus. Since PAR-2 mRNA was detected in highly vascularized organs, we compared the physiological functions of the thrombin receptor and PAR-2 in vascular endothelium. Thrombin and trypsin both elicited endothelium-dependent relaxations in prostaglandin F2alpha (PGF2alpha)-contracted strips of porcine coronary artery. Whereas high doses of both thrombin or trypsin (10 U/mL) caused homologous desensitization, trypsin caused further relaxation of thrombin-desensitized tissues. Thrombin and PAR-2-derived peptides (SFLLRN and SLIGRL) both induced endothelium-dependent relaxations in PGF2alpha-contracted porcine coronary arteries. SFLLRN or SLIGRL (30 micronmol/L) also showed homologous desensitization but not cross desensitization. In the presence of the NO synthase inhibitor NG-monomethyl-L-arginine (1 mmol/L), both SFLLRN- and SLIGRL-induced relaxations were partially inhibited. SFLLRN elicited weak contraction in coronary arteries without endothelium, whereas SLIGRL had no effect. Intravenous injection of SFLLRN (1 mg/kg, bolus) into anesthetized rats elicited a transient depressor response followed by pronounced pressor response. In contrast, intravenous administration of SLIGRL (1 mg/kg, bolus) produced only a marked depressor response. Consistent with the in vivo data, SFLLRN contracted the endothelium-rubbed rat aortic rings and aggregated human platelets in vitro, whereas SLIGRL had no effect. The finding that both trypsin and SLIGRL induced endothelium-dependent relaxations indicates the presence of PAR-2 on endothelial cells. In addition, both trypsin and SLIGRL elicited relaxations in thrombin- or SFLLRN-desensitized tissue, suggesting that PAR-2 is distinct from thrombin receptor in vascular endothelium. The lack of PAR-2-mediated platelet aggregation or smooth muscle contraction suggested it might not share the pathogenic properties associated with the thrombin receptor in the vasculature.
Animals↗
Medication nebulizer performance. Effects of diluent volume, nebulizer flow, and nebulizer brand.
BACKGROUND: Medication nebulizers are commonly used to delivery aerosolized medications to patients with respiratory disease. We evaluated output and respirable aerosol available to the patient (inhaled mass) for 17 medication nebulizers using a spontaneous breathing lung model. METHODS: Three nebulizer fill volumes (3, 4, and 5 mL containing 2.5 mg of albuterol) and 3 oxygen flows (6, 8, and 10 L/min) were evaluated using the 17 nebulizers. A cotton plug at the nebulizer mouthpiece was used to trap aerosol during simulated spontaneous breathing. Following each trial, the amount of albuterol remaining in the nebulizer and the amount deposited in the cotton plug were determined spectrophotometrically. Aerosol particle size was determined using an 11-stage cascade impactor. RESULTS: Increasing fill volume decreased the amount of albuterol trapped in the dead volume (p < 0.001) and increased the amount delivered to the patient (p < 0.001). Increasing flow increased the mass output of particles in the respirable range of 1 to 5 microns (p = 0.004), but the respirable mass delivered to the patient was affected to a greater extent by nebulizer brand (p < 0.001) than flow. Although 2.5 mg of albuterol was placed into the nebulizers, less than 0.5 mg in the respirable range of 1 to 5 microns was delivered to the mouthpiece. CONCLUSIONS: The performance of medication nebulizers is affected by fill volume, flow, and nebulizer brand. When they are used for research applications, the nebulizer characteristics must be evaluated and reported for the conditions used in the investigation.
Administration, Inhalation↗
Psychic reality: theoretical concepts.
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Ego↗
Quorum sensing: a population-density component in the determination of bacterial phenotype.
In bacteria, a network of cellular transduction mechanisms integrate signals from the bacterial environment to control gene expression, and thereby the bacterial phenotype. 'Quorum sensing' describes one such signalling mechanism in response to population density. It relies on the accumulation of small extracellular signalling molecules to modulate the transcription of target operons.
Anti-Bacterial Agents↗
Goodbye amalgam, hello alternatives?
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Alloys↗
A clinical and interactionist perspective on the bully-victim-bystander relationship.
The authors propose a dialectical conceptual framework for the bully-victim-bystander interaction often seen in school violence. A clinical and interactional typology is proposed for the detection of children falling into these patterns, with the goal of affording early and vigorous intervention and consultation opportunities with teachers and school administrators.
Adolescent↗
Analysis of bacterial carbapenem antibiotic production genes reveals a novel beta-lactam biosynthesis pathway.
Carbapenems are beta-lactam antibiotics which have an increasing utility in chemotherapy, particularly for nosocomial, multidrug-resistant infections. Strain GS101 of the bacterial phytopathogen, Erwinia carotovora, makes the simple beta-lactam antibiotic, 1-carbapen-2-em-3-carboxylic acid. We have mapped and sequenced the Erwinia genes encoding carbapenem production and have cloned these genes into Escherichia coli where we have reconstituted, for the first time, functional expression of the beta-lactam in a heterologous host. The carbapenem synthesis gene products are unrelated to enzymes involved in the synthesis of the so-called sulphur-containing beta-lactams, namely penicillins, cephamycins and cephalosporins. However, two of the carbapenem biosynthesis genes, carA and carC, encode proteins which show significant homology with proteins encoded by the Streptomyces clavuligerus gene cluster responsible for the production of the beta-lactamase inhibitor, clavulanic acid. These homologies, and some similarities in genetic organization between the clusters, suggest an evolutionary relatedness between some of the genes encoding production of the antibiotic and the beta-lactamase inhibitor. Our observation are consistent with the evolution of a second major biosynthetic route to the production of beta-lactam-ring-containing antibiotics.
Amino Acid Sequence↗
Design of multiple binary outcome studies with intentionally missing data.
We discuss the design and analysis of studies involving multiple binary outcomes in which only a subset of these outcomes can be measured on each individual. Such studies with "intentionally missing data" may arise due to practical or economic constraints; several examples from toxicology serve as illustrations. A global test statistic based on generalized estimating equations is presented and evaluated under a variety of missing patterns and correlation structures. Extensions of the global test statistic to allow for clustered data are also described. The relative efficiency of the global test statistic with missing data relative to that for complete data is investigated, both under a common dose effect alternative and when exposure has differential effects on the multiple endpoints. The implications of these efficiency calculations on study design are explored, and several recommendations are provided.
Animals↗
Rapid tryptic mapping using enzymatically active mass spectrometer probe tips.
A method has been developed for rapid, sensitive, and accurate tryptic mapping of polypeptides using matrix-assisted laser desorption/ionization time-of-flight mass analysis. The technique utilizes mass spectrometer probe tips which have been activated through the covalent immobilization of trypsin. The enzymatically active probe tips were used for the tryptic mapping of chicken egg lysozyme and the results compared with those obtained using either free trypsin or agarose-immobilized trypsin. A significant increase in the overall sensitivity of the process was observed using the active probe tips, as well as the production of more characteristic proteolytic fragments and the elimination of background signals due to the autolysis of the trypsin. Further, probe tip digestions were found to be rapid and convenient.
Amino Acid Sequence↗
Characterization of five type II arabinogalactan-protein fractions from red wine of increasing uronic acid content.
Five arabinogalactan-protein conjugates (AGP) were separated from red wine by two successive anion-exchange chromatography steps and further purified to apparent homogeneity by affinity and size-exclusion chromatography. Together they represent more than 40% of total wine polysaccharides, confirming the abundance of AGPs in red wine. The five purified fractions had a common arabinogalactan core with characteristics typical of wine type II AGPs, but differed mainly in their uronic acid content, as evidenced by differences in the strength of their binding to the anion-exchanger. Their uronic acid content and glycosidic linkage composition revealed that the three less acidic AGPs contained from 3 to 7% glucuronic acid, half in terminal non-reducing positions and half in terminal Rhap-(1-->4)-Glc pA-(1-->sequences. The two more acidic AGP-containing fractions contained both glucuronic (6.1 and 13.3%, respectively) and galacturonic (1.9 and 2.3%, respectively) acid in association with 2- and 2,4-linked rhamnose, indicating the presence of AG-rhamnogalacturonan fragments.
Carbohydrate Conformation↗
Plasmid DNA entry into postmitotic nuclei of primary rat myotubes.
These studies were initiated to elucidate the mechanism of DNA nuclear transport in mammalian cells. Biotin- or gold-labeled plasmid and plasmid DNA expression vectors for Escherichia coli beta-galactosidase or firefly luciferase were microinjected into the cytoplasm of primary rat myotubes in culture. Plasmid DNA was expressed in up to 70% of the injected myotubes, which indicates that it entered intact, postmitotic nuclei. The nuclear transport of plasmid DNA occurred through the nuclear pore by a process common to other large karyophilic macromolecules. The majority of the injected plasmid DNA was sequestered by cytoplasmic elements. This understanding of plasmid DNA nuclear transport provides a basis for increasing the efficiency of gene transfer.
Animals↗
Suramin suppresses hypercalcemia and osteoclastic bone resorption in nude mice bearing a human squamous cancer.
Suramin is a polyanionic agent which has been found to be an effective antineoplastic agent against various human tumors including adrenal, renal and prostatic cancer, and osteosarcoma. Recently, suramin has been shown to inhibit bone resorption in organ cultures of mouse calvarial bones. In the present study, we examined the effects of suramin on increased osteoclastic bone resorption and hypercalcemia in nude mice bearing a human oral squamous carcinoma. Suramin (1 mg/mouse/injection) was administered i.p. three times a week for the first 2 weeks and then once weekly for the next 6 weeks. Blood ionized calcium levels in the suramin-treated cancer-bearing group were significantly lower than those in the untreated cancer-bearing group. Histological and histomorphometrical examination of bones of these animals showed a significant decrease in osteoclast numbers in the suramin-treated cancer-bearing animals. Suramin at a dose of 0.1 mg/mouse/injection was ineffective and 2 mg/mouse/injection was toxic, confirming its narrow effective dose. Suramin showed no effects on the growth of this squamous cancer. However, suramin markedly inhibited in vivo growth of a rat prostatic adenocarcinoma. In mouse marrow cultures, suramin decreased osteoclast-like cell formation in a dose-dependent manner. Furthermore, suramin also inhibited bone resorption in organ cultures of fetal rat long bones and resorption pit formation by isolated mature rat osteoclasts. These results show that suramin is an effective inhibitor of osteoclastic bone resorption in vitro and in vivo and suggest that suramin may be a useful agent in prevention and treatment of cancer-induced hypercalcemia. However, our results also suggest that for this indication suramin has a confined range of effective dose.
Adenocarcinoma↗
Mass spectrometric immunoassay.
A new, general method of immunoassay is demonstrated. The approach is based on the microscale immunoaffinity capture of target antigens followed by mass-specific identification and quantitation using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Immunoaffinity capture of antigens effectively overcomes signal suppression effects typically encountered during traditional matrix-assisted laser desorption/ionization analysis of complex biological mixtures while simultaneously concentrating the analyte into a small volume. Mass spectrometric detection of antigens is unambiguous, as antigen signals are observed at characteristic mass-to-charge values in the mass spectrum, offering a high level of immunity to artifacts due to nonbiospecific retention of mixture components. However, the most important aspect of such mass-specific detection is the ability to use a single assay to screen biological systems for the presence of multiple, mass-resolved antigens. Analyte quantitation is possible by using a single antibody to capture both the antigen and an antigen variant which has been chemically modified to have a different mass. With proper calibration, the relative signal intensities of the two species in the mass spectrum can be used to determine the antigen concentration. Sample incubation and processing methods were such that a typical analysis could be performed in less than 1 h while subnanomolar sensitivities were maintained. The technique has been used for the rapid, selective, and quantitative screening of human blood for the presence of myotoxin a, and Mojave toxin form the venoms of the prairie rattlesnakes, Crotalus viridis viridis, and and the Mojave rattlesnake, Crotalus scutulatus scutulatus.
Animals↗
Platinum anticancer drug binding to DNA detected by thickness-shear-mode acoustic wave sensor.
Nucleic acid has been attached to the electrodes of thickness-shear-mode acoustic wave devices to produce a biosensor for platinum-based drugs. The decreases in series resonant frequency for interactions of DNA with both cis- and transplatin are indicative of two distinct kinetic processes. The results of a kinetic analysis are interpreted in terms of nucleic acid binding of the hydrolysis products of the two drugs. Concentration-dependent decreases of series resonant frequency show that the limit of detection for the drugs is approximately 10(-7) M. Motional resistance changes for nucleic acid-drug interactions also convey information regarding the chemistry of the macromolecules at the interface.
Animals↗
Oxidation of low-density lipoproteins: effect of antioxidant content, fatty acid composition and intrinsic phospholipase activity on susceptibility to metal ion-induced oxidation.
The oxidative modification of low-density lipoprotein (LDL) may play an important role in atherogenesis. Our understanding of the mechanism of LDL oxidation and the factors that determine its susceptibility to oxidation is still incomplete. We have isolated LDL from 45 healthy individuals and studied the relationship between LDL fatty acid, vitamin E and beta-carotene composition, intrinsic phospholipase A2-like activity and parameters of LDL oxidation. LDL was exposed to a copper ion-dependent oxidising system and the kinetics of oxidation studied by monitoring formation of fatty acid conjugated dienes. The length of the lag phase of inhibited lipid peroxidation was measured as well as the rate of lipid peroxidation during the propagation phase. There was no significant correlation between LDL antioxidant vitamin or fatty acid composition and lag time to LDL oxidation. Oleic acid was negatively correlated with the rate of LDL oxidation (r = -0.41, P < 0.01) whilst linoleic acid was significantly correlated with the extent of LDL oxidation measured by the production of total dienes (r = 0.34, P < 0.05). Interestingly, LDL vitamin E content was positively correlated with both the rate (r = 0.28, P < 0.05) and extent of LDL oxidation (r = 0.43, P < 0.01). LDL isolated from this group of subjects showed significant intrinsic phospholipase-like activity. The phospholipase activity, whilst not correlated with lag time, was significantly correlated with both rate (r = 0.43, P < 0.01) and total diene production (r = 0.44, P < 0.01) of LDL oxidation. We conclude that antioxidant content, fatty acid composition and intrinsic phospholipase activity have little influence on the lag time of Cu-induced LDL oxidation. These components do however, significantly influence both the rate and extent of LDL oxidation, with increased vitamin E, linoleic acid content and phospholipase activity associated with faster and more extensive oxidation. The possible pro-oxidant effect of vitamin E has interesting implications for the postulated 'protective' effects of vitamin E on atherogenesis.
Antioxidants↗
Detection of human IgM at m/z approximately 1 MDa.
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Humans↗
Peptide characterization using bioreactive mass spectrometer probe tips.
A method has been developed for the rapid and sensitive mass spectrometric characterization of peptides. The approach uses bioreactive mass spectrometer probe tips, incorporating covalently bound enzymes, which are capable of modifying biomolecules for analytical purposes. In the demonstrated cases, enzymatic proteolysis is initiated upon application of analyte to the probe tips, time is allowed for digestion, and the products are analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The probe tips have been used for proteolytic mapping and partial sequence determination of picomole quantities of peptide. Analysis times were approximately 30 min. Two methods of database search were utilized. The first used limited peptide sequence information and parent molecular weight, while the second used exclusively the molecular weights of a number of endoproteolytic fragments. A simple method of comparing the match of experimental data for a tryptic digest with the results of a search is described.
Amino Acid Sequence↗