Do cardiologists have higher thresholds for recommending coronary arteriography than family physicians?

The purpose of this study was to use a new model of decision making to understand variability in physicians' utilization of diagnostic tests. We studied physicians' recommendations for coronary arteriography in hypothetical patients with chest pain by analyzing responses of 235 cardiologists and family physicians. Thresholds for testing were derived by obtaining estimates of the probability of disease and recommendations for coronary arteriography before and after an exercise test.

The importance of considering single testing when two tests are available.

When considering two dichotomous tests in combination for reaching a treatment decision, the choice between single and multiple testing depends, in part, on the pretest probability of disease. The authors show that two tests are never preferred to a single test for all disease probabilities, regardless of whether the two tests are performed in parallel or in series.

Clinical reasoning and cognitive processes.

Expected utility theory, and the Bayesian probability theory on which it is based, form the normative basis of most work in medical decision analysis. Recent work in the psychology of judgments and decisions indicates that people do not conform to the axioms of this theory and that these deviations occur in clinical reasoning as well as in the psychology laboratory. At issue is what to do now.

The protein synthesis initiation factor 2 G-domain. Study of a functionally active C-terminal 65-kilodalton fragment of IF2 from Escherichia coli.

Protein synthesis initiation factor 2 (IF2) is present in Escherichia coli cells as two forms which are expressed from the same gene: IF2 alpha [97.3 kilodaltons (kDa)] and IF2 beta (79.7 kDa).

Translational repression by chemical inducers of the stress response occurs by different pathways.

The mechanism by which chemical inducers of the stress response inhibit protein synthesis was examined. All the chemicals tested principally inhibit the initiation phase of translation. Covalent modification of the initiation factor proteins does not constitute a common mechanism.

Cloning and mapping of infA, the gene for protein synthesis initiation factor IF1.

The gene for translation initiation factor IF1, infA, has been identified by using two synthetic oligonucleotides to screen a Charon 30 library of Escherichia coli DNA. A recombinant lambda phage, C1921, was purified from a plaque positive for both probes. A 2.

The effect of neodymium:YAG laser shocks on the blood-aqueous barrier.

An animal model demonstrates that the acute inflammation seen after neodymium: YAG (Nd:YAG) capsulotomy is related to the presence of disrupted tissue suspended in the aqueous, rather than to the mechanical insults by the repeated shock waves. Seven rabbits were treated in the lens cortex of one eye with 20 bursts of 4 pulses, 24 mJ each, and followed fluorophotometrically using albumin labelled with fluorescein. This allowed transmission of shock waves to the anterior segment without releasing debris in five eyes that showed no inflammation.

Initiation factor protein modifications and inhibition of protein synthesis.

The protein covalent modification state of eucaryotic initiation factors eIF-2 and eIF-4B in HeLa cells was examined after they were exposed to a variety of conditions or treatments that regulate protein synthesis. A few factors (e.g.

Cloning and sequencing of complementary DNAs encoding the alpha-subunit of translational initiation factor eIF-2. Characterization of the protein and its messenger RNA.

A clone encoding the alpha-subunit of eukaryotic initiation factor 2 (eIF-2 alpha) was isolated from a lambda gt11 expression library of rat brain cDNAs. The fusion protein expressed by the recombinant phage reacts with eIF-2 alpha antiserum except when the serum is preadsorbed with pure eIF-2. The translation of hybrid-selected HeLa cell mRNA produces two proteins which are indistinguishable from authentic HeLa eIF-2 alpha and its phosphorylated form when analyzed by electrophoresis in two-dimensional isoelectrofocusing/sodium dodecyl sulfate-polyacrylamide gels and by partial protease digestion.

Regulation of polypeptide chain initiation in Chinese hamster ovary cells with a temperature-sensitive leucyl-tRNA synthetase. Changes in phosphorylation of initiation factor eIF-2 and in the activity of the guanine nucleotide exchange factor GEF.

When cultures of the temperature-sensitive Chinese hamster ovary cell mutant tsH1 are shifted from 34 degrees C (permissive temperature) to 39.5 degrees C (nonpermissive temperature), protein synthesis is inhibited by more than 80%. This is due principally to a block in activity of polypeptide chain initiation factor eIF-2.

Regulated phosphorylation and low abundance of HeLa cell initiation factor eIF-4F suggest a role in translational control. Heat shock effects on eIF-4F.

Initiation factor eIF-4F, a multiprotein cap binding protein complex, was purified from HeLa cells by m7G affinity chromatography and independently by phosphocellulose column chromatography. The m7G affinity-purified sample contains three major proteins, p220, eIF-4A, and p28 (also known as CBP-I or eIF-4E). The abundancies of these proteins are roughly 2, 10, and 0.

Translational initiation factors from sea urchin eggs and embryos: functional properties are highly conserved.

We have used three mammalian in vitro assays for translational initiation (globin synthesis, methionyl-puromycin synthesis, and ternary complex formation), consisting of defined components, to ask whether sea urchin (Strongylocentrotus purpuratus) egg and embryo translational components are active in heterologous assays for mammalian components, and to determine to what extent these activities are evolutionarily conserved. A "pH 5 enzyme" fraction prepared from unfertilized eggs and embryos efficiently replaced the rat liver pH 5 fraction in a globin synthesis assay, indicating that the elongation and termination factors and the aminoacyl-tRNAs are compatible with the mammalian translational machinery. The classical schemes for mammalian initiation factor purification yielded low or no detectable activities in the ribosomal salt washes, so a novel procedure was developed to partially purify initiation factors from sea urchin eggs and embryos before testing for activity.

Changes in eIF-4D hypusine modification or abundance are not correlated with translational repression in HeLa cells.

Initiation factor eIF-4D is represented by about 11 X 10(6) molecules/HeLa cell (0.45% of the cytoplasmic protein molecules). The fraction of eIF-4D that contains the post-translational modification of lysine converted to hypusine is not regulated with respect to translation rate in HeLa cells.

Binding of Escherichia coli protein synthesis initiation factor IF1 to 30S ribosomal subunits measured by fluorescence polarization.

The interaction of initiation factor IF1 with 30S ribosomal subunits was measured quantitatively by fluorescence polarization. Purified IF1 was treated with 2-iminothiolane and N-[[(iodoacetyl)-amino]ethyl]-5-naphthylamine-1-sulfonic acid in order to prepare a covalent fluorescent derivative without eliminating positive charges on the protein required for biochemical activity. The fluorescent-labeled IF1 binds to 30S subunits and promotes the formation of N-formylmethionyl-tRNA complexes with 70S ribosomes.

Interaction between initiator Met-tRNAfMet and elongation factor EF-Tu from E. coli.

It has recently been shown that the non-formylated initiator Met-tRNAfMet from E. coli can form a stable ternary complex with the elongation factor EF-Tu and GTP. Using the protection of EF-Tu:GTP against spontaneous hydrolysis of the aminoacylester bond of Met-tRNAfMet, we confirm these results, and show that the protection is specific for the non-formylated form of the initiator tRNA.

Multiple test analyzer (MTA): a microcomputer program for determining preferred strategies with two diagnostic tests.

There are ten distinct management strategies in clinical situations that involve two diagnostic tests with dichotomous outcomes. The authors describe a microcomputer program, based on a previously described model, that can be used to identify test and test-treatment thresholds and to compute preferred strategies. The program provides tables and graphs of the results, which can be viewed or printed, and there is an optimization routine that facilitates comprehensive analysis.

Clinical guidelines for using two dichotomous tests.

Building on the threshold model developed by Pauker and Kassirer for a single test, the authors describe a decision analytic model for two tests with dichotomous outcomes. The model includes ten decision strategies that differ depending on which tests are performed, whether the tests are performed together or in sequence, and the definition of a positivity criterion used to make the treatment decision when the test results disagree. Formulas derived from the model are used to compute the preferred option as a function of disease probability and to calculate test and test-treatment thresholds.

Comparing aggregate estimates of derived thresholds for clinical decisions.

Thresholds for medical decision making are the probabilities of disease at which clinicians choose to initiate testing or therapy. A descriptive analysis of clinicians' decision making can derive their test and test-treatment thresholds and has the potential to explain variations in test utilization. A previously described method summarizes thresholds for a group of clinicians by determining the range of probability which includes the maximum number of clinicians' individual thresholds.

Overproduction and purification of initiation factor IF2 and pNUSA proteins from a recombinant plasmid bearing strain.

The genes for translational initiation factor, IF2 and pNusA have been cloned into a plasmid vector where they are placed under the control of the inducible lambdapL promoter and the c1857 thermosensitive repressor. When a strain carrying this plasmid is heat induced, IF2 alpha, IF2 beta and pNusA are overproduced 15 to 20 fold. This has allowed us to purify the IF2 and NusA proteins in large amounts.

Double-stranded RNA-dependent protein kinase and 2-5A system are both activated in interferon-treated, encephalomyocarditis virus-infected HeLa cells.

Activation of the interferon-inducible, double-stranded RNA-dependent protein kinase was monitored in monolayer cultures of control and interferon-treated HeLa cells infected with encephalomyocarditis virus. The extent of phosphorylation in the intact cell of the alpha-subunit of eucaryotic protein synthesis initiation factor eIF2 by the kinase was determined for the first time in this type of system, using a two-dimensional immunoblot technique. Virus protein synthesis and the kinetics of activation of the ppp(A2'p)nA (n greater than or equal to 2) system were analyzed in parallel.

Regulation of initiation factors during translational repression caused by serum depletion. Abundance, synthesis, and turnover rates.

During growth in unreplenished medium, the fraction of active, polysomal ribosomes progressively decreases about 3-fold from 80-90% to only 20-40% due to a reduced rate of initiation. To assess whether the abundance of initiation factors could be involved in this repression of translational activity. HeLa cell cytoplasmic lysates were resolved by two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and spots corresponding to the initiation factor proteins were quantitated.

Regulation of initiation factors during translational repression caused by serum depletion. Covalent modification.

One to 2 h after transfer of HeLa cells into fresh serum-containing medium, when translation rates are maximal, the initiation factor proteins were examined on immunoblots of two-dimensional gels. Eukaryotic initiation factor (eIF)-2 alpha, eIF-2 beta, and eIF-4A each formed a single immunoreactive spot; eIF-2 gamma formed 2 spots; and eIF-4B formed a complex array of 12-20 spots. After 4 days of growth in unreplenished medium, when translation rates have dropped 4-6-fold, several alterations in the isoelectric forms were observed: eIF-2 alpha now occurred in 2 forms, eIF-2 beta was present in 3-4 forms, and the most acidic cluster of eIF-4B variants was decreased or absent while a new isoelectric variant appeared at the basic end of the array.

Shutoff of host translation by encephalomyocarditis virus infection does not involve cleavage of the eucaryotic initiation factor 4F polypeptide that accompanies poliovirus infection.

Studies were conducted to determine whether encephalomyocarditis virus infection causes proteolytic cleavage of any of the polypeptides which comprise eucaryotic initiation factor 4F. Since no such alterations in the components of the initiation factor were detected, these observations confirmed that the mechanisms whereby encephalomyocarditis virus and poliovirus shut off host translation are different.