Study of the two pathways for arachidonate oxygenation in blood platelets

During collagen-induced blood platelet aggregation, arachidonic acid is set free from membrane phospholipids and subsequently converted into 12-hydroxyeicosatetraenoic acid by arachidonate lipoxygenase and into thromboxane A₂, 12-hydroxyheptadecatrienoic acid (HETE) and malondialdehyde by cyclooxygenase and thromboxane synthase. Lipoxygenase and cyclooxygenase have optimal activity at neutral to basic pH, while the thromboxane synthase is pH-independent between 5 and 9. These enzymes are membrane-bound. The cyclooxygenase is rapidly inactivated upon membrane disruption by nonionic detergents or phospholipid degradation with phospholipase A₂. It was found that platelet phospholipase A₂ preferentially splits off fatty acid with four double bonds. Eicosatetraynoic acid was used to investigate the physiological function of the arachidonate lipoxygenase during collagen-induced aggregation of rat blood platelets. This fatty acid is a more efficient inhibitor of lipoxygenase than of cyclooxygenase. At an inhibitor concentration of 0.6 μg/ml, platelet aggreation, 12-hydroxyeicosatetraenoic acid production as well as 15-hydroxytryptamine release are completely inhibited, while there is an apparent stimulation of the cyclooxygenase. These results indicate that arachidonate lipoxygenase is essential for irreversible blood platelet aggregation.     

Influence of multiple models on the behavior of institutionalized retarded children: increased generalization to other models and other behaviors

After being pretested to determine base levels of imitation, 32 9-14 yr old retarded children were reinforced for imitating a model in 9 training sessions. Ss in a single model condition were reinforced by the same model across all sessions, whereas Ss in a multiple model condition were reinforced by 3 different models (3 sessions per model). A posttest to assess levels of imitation was then conducted by a model with whom the Ss had not had contact and who demonstrated a new set of behaviors. Results during training sessions show that (a) Ss learned to imitate, and this learning was not inhibited by multiple models; and (b) Ss generalized and imitated nonreinforced behaviors, and this response generalization was facilitated by multiple models. Most importantly, pre-posttest comparisons indicated that generalized use of the new response class (imitation) with new models was 8 times greater for Ss trained with multiple as opposed to single models. Implications for the maintenance and generalized effectiveness of social intervention programs are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Serologic response to revaccination with two rubella vaccines

Three years after receiving rubella vaccine, 1,060 elementary school children living on the island of Maui, Hawaii, were revaccinated with either HPV-77 DE-5 or RA 27/3 rubella vaccine given subcutaneously or intranasally in order to compare the effectiveness of these two vaccines in raising antibody titers. RA 27/3 was the more effective booster vaccine, producing fourfold or greater titer rises in 20.1% of recipients, including 80% of children with hemagglutination-inhibiting antibody titers less than or equal to 1:40 at the time of revaccination, intranasal revaccination was not significantly more effective than subcutaneous revaccination, although it did elicit higher titers in children who responded. Responses differed according to the vaccine that children had received three years earlier. Because antibody titers have persisted in vaccinated children, routine administration of a second dose of rubella vaccine is not currently recommended.     

Cross-linking studies and membrane localization and assembly of radiolabelled large mechanosensitive ion channel (MscL) of Escherichia coli

While the overall energy landscape of a foldable protein can be described by means of a few parameters characterizing its statistical topography, specific energetic terms subtly bias the representative structures giving rise to residue pair correlations as in a liquid. We use a free energy functional incorporating an inhomogeneous pair contact energy along with a contact formation entropy and a cooperativity contribution to determine residue-specific contact probabilities in the denatured state and the transition state ensemble. The predicted "hot residues" for the theoretical transition state ensemble reasonably agree with experiment for chymotrypsin inhibitor 2, and generally a strong correlation exists with the measured kinetic effects of mutating residues not involved in highly solvent-exposed regions.     

p53 mutation spectrum in relation to GSTM1, CYP1A1 and CYP2E1 in surgically treated patients with non-small cell lung cancer

OBJECTIVE: The purpose of this study was to assess the feasibility of using a prototype split-specimen design to assess integrity of a portion of the total testing process in medical clinics and laboratories. DESIGN: Two or three tubes of venous blood were collected from 177 patients for analysis of one of three analytes (serum potassium, serum total cholesterol, and whole-blood hemoglobin). Patients were seen at one of the nine clinics participating in this study. In all cases, one tube of blood from each patient was sent to a commercial referral laboratory, and the other tube(s) forwarded to the laboratory that routinely tested specimens for the clinic (participating laboratory) for analysis. Each participating laboratory removed a preanalysis and sometimes a post-analysis aliquot from each specimen and forwarded these to the referral laboratory for analysis. SETTING: The study was conducted in six physician office laboratories (three serving 1 to 4 [mean, 2.7] internists and three serving 3 to 24 [mean, 12] family physicians) and three hospital laboratories (serving hospitals with 100 to more than 700 beds). PATIENTS: Study patients were voluntary participants and provided informed consent. Patient age ranged from 18 to 80 years, and for all the laboratory test was specifically ordered for clinical reasons. Patients who were unable or unwilling to provide informed consent, those for whom testing would require that they provide more than 100 mL of blood, those whose blood was being collected by fingerstick, and those with results that were part of a laboratory test profile were excluded. MAIN OUTCOME MEASURES: Two main outcome measures were assessed: (1) percent differences between split-specimen results exceeding the maximum allowable imprecision level, which was based on published biological variation data (defined as one-half of the intraindividual percent coefficient of variation), for each analyte (result discrepancies); and (2) all "problems" (defined as departures from standard operating procedures) that could be documented by retrospective review of all relevant medical and laboratory records. RESULTS: The rate of result discrepancies was 1 in 20 (5%) for patients in whom hemoglobin was analyzed, 12 in 57 (21%) for patients in whom potassium was analyzed, and 1 in 60 (2%) for patients in whom total cholesterol was analyzed. Results of samples obtained during the aliquoting and storage phases of the total testing process were subject to study-induced problems and were generally not useful in tracing problems to specific stages of the testing process. A total of 28 problems (involving 26 patients) were documented, but only 6 problems were due to routine testing processes. CONCLUSIONS: The feasibility and limitations of a split-specimen design to detect result discrepancies were demonstrated. Most documented problems (22 of 28, or 79%) were study induced. To assess integrity of the total testing process, such problems need to be avoided.     

Stability effects of increasing the hydrophobicity of solvent-exposed side chains in staphylococcal nuclease

A total of fifty single site surface phenylalanine substitution mutants have been made in the model protein staphylococcal nuclease. The fifty residues that were replaced with phenylalanine were chosen to give a broad sampling of solvent accessibility, secondary structure, and backbone conformations. The change in the stability of each mutant protein relative to wild type was measured by guanidine hydrochloride denaturation. These results were compared to previous results obtained when these same sites were substituted with an alanine and a glycine. By this means, changes in the stability due to the loss of interactions of the wild-type side chain can be separated from the effects of introducing the bulky, hydrophobic phenylalanine in these solvent-exposed positions. In general, our results agree with the conventional wisdom that placing a hydrophobic residue in a solvent-exposed position is destabilizing in most cases, but less destabilizing than most changes in the hydrophobic core of the protein. However, the degree to which a hydrophobic surface substitution destabilizes or stabilizes a globular protein is highly context-dependent, with some mutations being as destabilizing as those in the core. This indicates that steric and packing considerations are also important on the surface of a globular protein but generally are not as important as in the interior. No evidence for the widespread occurrence of the so-called reverse hydrophobic effect at solvent-exposed sites was found. In addition, this survey of numerous sites suggests that previous measurements of alpha-helix "propensities" often seriously underestimate the importance of the environment of the side chain.     

Priming of platelet alphaIIbbeta3 by oxidants is associated with tyrosine phosphorylation of beta3

Reactive oxygen species play an important role at the site of vascular injuries and arterial thromboses. We studied the mechanism mediating platelet aggregation induced by H2O2, a major cellular oxidant. Exposure to H2O2 triggered platelet aggregation, but only when the platelets were stirred. Strong platelet aggregation induced99032416 required the presence of the tyrosine phosphatase inhibitor sodium orthovanadate (NaVO4) and was dependent on the participation of integrin alphaIIbbeta3 (glycoprotein IIb-IIIa). A specific inhibitor of alphaIIbbeta3 blocked platelet aggregation induced by H2O2 and NaVO4, thus confirming that aggregation requires this receptor. In the presence of H2O2 and NaVO4, multiple platelet substrates were phosphorylated on tyrosine. Such tyrosine kinase response was necessary but not sufficient to activate alphaIIbbeta3, as detected by binding of soluble fibrinogen to platelets. Stirring of the platelets exposed to H2O2 and NaVO4 was also needed to allow for binding of fibrinogen to alphaIIbbeta3. The tyrosine kinase inhibitor genistein was able to block platelet aggregation induced by H2O2 and NaVO4, thus confirming that tyrosine kinase activity was needed to trigger alphaIIbbeta3 activation on stirring. N-Acetyl-L-cysteine, a cell-permeant antioxidant, blocked the tyrosine phosphorylation of platelet substrates and also the platelet aggregation induced by H2O2 and NaVO4. We found that beta3 was phosphorylated on tyrosine in platelets exposed to H2O2 and NaVO4, even in the absence of aggregation. Hence, tyrosine phosphorylation of beta3 might contribute to the "priming" of alphaIIbbeta3 induced by H2O2 and NaVO4, whereby the receptor can become activated on stirring of the platelets.     

The measurement of the lateralization of narrow bands of noise using an acoustic pointing paradigm: the effect of sound-pressure level

The effects of varying interaural time delay (ITD) and interaural intensity difference (IID) were measured in normal-hearing subjects as a function of eleven frequencies and at sound-pressure levels (SPL) from 60 to 90 dB SPL and at 25-dB sensation level. Using an "acoustic" pointing paradigm, the IID of a 500-Hz narrow-band (100 Hz) noise (the "pointer") was varied by the subject to coincide with that of a "target" ITD stimulus. ITDs of 0, +/- 200, and +/- 400 microseconds were obtained through total waveform delays of narrow-band noise (NBN), including envelope and fine structure. The results of this experiment confirm the traditional view of binaural hearing for like stimuli: There is little perceived displacement away from 0 IID at frequencies of 1250 Hz and above. In the low frequencies, subjects required IIDs greater than the expected 10 dB to perceive a fully lateralized image, and they varied in the maximum value of IID that they required, regardless of frequency. Our subjects did not always perceive the intracranial locations of ITD targets symmetrically: When the signal was delayed to one ear, the resultant matching IID was often different in magnitude than for the same ITD target delayed to the opposite ear for the identical frequency. The results of two subjects suggested that people with asymmetric normal hearing have adapted to their asymmetry for lateralization tasks: The subjects were found to lateralize toward the ear with the greater SPL stimulus, regardless of the ear to which the signal was delayed, when signals of equal SL were presented, and toward the leading ear when signals of equal SPL were presented (unequal SL). Increasing the presentation levels above 60 dB SPL had an effect on the perception of high-frequency ITD targets: As the intensity level increased, the slopes of the IID versus ITD functions increased indicating better discrimination of ITD. This study is in agreement with other studies in providing strong evidence of individual differences in lateralization experiments. These individual differences might be attributable to differential sensitivity to ambiguous time stimulus cues, differential task sensitivity, age effects, threshold asymmetries, or criterion variability.