Transcript analysis and comparative evaluation of shaker and slowmo gene homologues from the European corn borer, Ostrinia nubilalis.

The movement and dispersal of larval Lepidoptera impact their survival and distribution within the natural landscape. Homologues of the Drosophila behaviour-linked genes shaker (shkr) and slowmo (slmo) were identified from Ostrinia nubilalis (Lepidoptera: Crambidae). Onshkr was isolated as a 1610-nucleotide (nt) constitutively expressed transcript encoding a membrane-localized 469-amino-acid (aa) protein with a conserved tetramerization domain and the six-domain architecture necessary for the molecule to fold into an active K(+) channel. Three expressed splice variants of 682, 970 and 1604 nt were identified for the Onslmo gene, and encode predicted 141 and 228 aa proteins with a conserved protein of relevant evolutionary and lymphoid interest (PRELI) domain that may function in mitochondrial protein sorting and perinuclear protein localization. Onshkr and Onslmo protein sequences aligned within monophyletic lepidopteran groups.

The locations of nonresidential out-of-hospital cardiac arrests in the City of Pittsburgh over a three-year period: implications for automated external defibrillator placement.

OBJECTIVES: To determine the locations of nonresidential out-of-hospital cardiac arrests (OHCAs) in the City of Pittsburgh and to determine whether there are "high-risk" locations that might benefit from placement of automated external defibrillators (AEDs). METHODS: This was a retrospective case review of paramedic calls for OHCA over a three-year period, in a mid-sized northeastern city. Cardiac arrests that were traumatic or for which the patients were dead on arrival (DOA) or had advanced directives for no resuscitation were excluded. Cardiac arrests that occurred in a public location (i.e., not a private residence) were categorized. RESULTS: A total of 971 OHCAs occurred in the City of Pittsburgh from January 1, 1997, to December 31, 1999. Of these, 575 (59%) occurred in private residences, and 396 (41%) occurred in nonresidential locations. Fifteen locations had at least one cardiac arrest per year for three years, accounting for 166 (43%) of the total nonresidential OHCAs. Twelve locations had two arrests during the three-year period, accounting for 24 (6%) of the total nonresidential OHCAs. One hundred ninety-four locations had a single episode of cardiac arrest, accounting for 51% of the OHCAs. Nursing homes and dialysis centers accounted for 178 (94%) OHCAs in the 27 locations that had two or more cardiac arrests. A local sports/events complex (Three Rivers Stadium) was the only other single location to have more than two cardiac arrests in the three-year study period, with a total of three. However, events at this complex are routinely staffed by paramedics equipped with defibrillators. CONCLUSION: The majority of nonresidential OHCAs occur as singular, isolated events. Other than nursing homes and dialysis centers, there were no identifiable high-risk locations for nonresidential OHCA within the City of Pittsburgh.

A word-ordering test to differentiate between aphasia and schizophrenia.

The capacity of a word-ordering task to differentiate aphasic and schizophrenic patients was assessed. Cards containing words from 10 types of sentences were given to the patients. Each sentence was presented individually with the words in jumbled order and patients were asked to reorder them to produce a gramatically correct sentence. Process and reactive schizophrenics were differentiated from aphasic patients using the dual criteria of more than three incorrect sentences or an average of more than 90 sec per sentence as indicative of aphasia. These criteria were initially tested on a group of aphasics matched with the schizophrenics on age and education and cross-validated using a second more representative sample (unmatched) of aphasics. Both comparisons showed no errors of classification. Further analyses indicated no need for separate norms or classification criteria related to the sex, age, or educational level of the patients.

Audiometric configuration in patients being treated for alcoholism.

The auditory acuity of 52 alcoholic subjects showed a consistent bilateral high frequency loss. Presence of this loss was found to be related to drinking time, but independent of age, suggesting a definite relationship between length of heavy drinking time and loss of auditory acuity in the high frequencies.

Binding of dipyridamole to human platelets and to alpha1 acid glycoprotein and its significance for the inhibition of adenosine uptake.

The interactions of dipyridamole with alpha(1) acid glycoprotein of plasma and with human platelets are related to inhibition of adenosine uptake by platelets. Binding studies by equilibrium gel filtration suggested that 1 mol of dipyridamole binds per mol of alpha(1) acid glycoprotein with a dissociation constant of 1.6 muM. Platelets contain two populations of binding sites, one with high and another with lower affinity for the drug. The binding of dipyridamole to the high-affinity sites follows a Michaelis-Menten binding pattern with a dissociation constant of 0.04 muM. Approximately 2 x 10(4) dipyridamole molecules are bound at the high-affinity sites of each platelet. The lower affinity sites bind the drug with a dissociation constant of 4 muM. In the presence of alpha(1) acid glycoprotein of plasma, the binding of dipyridamole to human platelets is inhibited. Correspondingly, the dipyridamole inhibition of adenosine uptake by platelets is reduced 1,000-fold by purified alpha(1) acid glycoprotein. The binding of dipyridamole to human platelets was found to be essential for its inhibition of adenosine uptake by platelets. Dipyridamole decreases the incorporation of [(14)C]adenosine radioactivity in platelet nucleotides and reduces the [(14)C]-ATP to [(14)C]ADP ratio. Purified alpha(1) acid glycoprotein reverses these effects of dipyridamole on adenosine metabolism of platelets in a concentration-dependent manner. An equilibrium of dipyridamole binding to alpha(1) acid glycoprotein and to platelets is proposed.

Metabolic aspects of the secretion of stored compounds from blood platelets. V. Effect of ionophore A23187 on washed platelets.

1. The A23187-induced secretion of preabsorbed serotonin from human blood platelets at 37 degrees C is studied. Preincubation at the same temperature before the addition of ionophore is necessary for maximal release induction. When total incubation time is kept constant, longer time with ionophore results in a smaller decrease in the level of metabolic ATP and increase in metabolic ATP and increase in metabolic IMP. This coincides with the reduction in secretion, but statistical treatment of the results suggests that the reduced secretion only partially explains the reduced drop in metabolic ATP, and that therefore a resynthesis of metabolic ATP from IMP may have taken place. 2. In some experiments induction of secretion takes place over a very narrow range of ionophore concentration. 3. When K+ substitutes for Na+ in the extracellular medium, the need for preincubation for maximal secretion becomes less evident, and at times is abolished, while there is still a significant increase in the metabolic ATP level by prolonged incubation with ionophore. 4. A reduction in secretion is observed with metabolic blockers when the ionophore is added after preincubation, but to a much less degree than when secretion is induced by thrombin, in spite of a great reduction in the level of metabolic ATP. This may partly be explained by the increase in secretion induction by A23187 in the presence of inhibitors when the ionophore is added in the cold, suggesting that the inhibitor may cause "weakening" of the platelets' "resistance" to induction of secretion by ionophore. 5. When the effect of Ca2+ and of Mg2+ on the level of intermediates of the TP leads to hypoxanthine conversion is studied, it is evident that the addition of Ca2+ causes enhanced IMP accumulation and a reduction in the level of inosine plus hypoxanthine, while Mg2+ has the opposite effect. This suggests that the two metals affect the enzymes of the IMP leads to hypoxanthine conversion differently. 6. Indomethacin inhibits secretion induced by A23187, suggesting that prostaglandin intermediates may amplify the ionophore-induced release. The adenine nucleotide metabolism is not affected. 7. The results indicate that there is an indirect, rather than direct, link between the major metabolic changes and the secretion induced by A23187, but that the ionophore may cause intracellular changes which are not connected to its effect as release inducer.

Calcium ionophore A23187 (Eli Lilly): effect on platelet function, structure and metabolism.

The addition of 0.1 muM ionophore A23187 to washed platelets incubated in citrated saline caused massive release of stored serotonin accompanied by intracellular accumulation of inosine monophosphate, but produced no detectable influx of externally added calcium or abnormal structural alterations. With increasing ionophore concentration there was a significant influx of calcium and a drastic alteration in the platelet ultrastructure. The increase in ionophore concentration was accompanied by the conversion of the major part of metabolic adenine nucleotides to inosine monophosphate and an almost complete blockage of further conversion to inosine and hypoxanthine. The metabolic changes were accentuated by the addition of calcium at concentrations less than 1/10 of the citrate concentration. In the presence of Ca++, or when citrate was omitted, there was a substantial leakage of cytoplasmic material, which at times suggested complete exchangeability between cytoplasm and extracellular medium. Our findings are consistent with the hypothesis that the platelet release reaction is triggered by intracellularly bound calcium. They also suggest that the application of high ionophore concentration has a toxicologic rather than a physiologic effect on platelets, and that a weak chelator added during incubation with the ionophore can in the absence of divalent cations prevent cell destruction, but not the toxic effect on cell metabolism.