Substance misuse among health care workers: national survey of occupational physicians.

BACKGROUND: National Health Service (NHS) occupational health departments assist in the identification and assessment of substance-use disorders among health care workers (HCWs) and are involved in the management of an individual's return to work after treatment. AIMS: To determine the experience and training of NHS occupational health physicians (OHPs) in identifying substance misuse among HCWs. METHODS: A national, cross-sectional, postal-based questionnaire was administered to the Association of National Health Occupational Physicians membership. RESULTS: A total of 145/224 (65%) OHPs (55% male), with a mean age of 49 years (SD ± 9.1; range 28-76), who had worked in the NHS on average for 9.6 years took part. The majority of respondents were consultant grade (59%). Since taking up their NHS post, 26% had received no training in substance misuse. Of those who had undergone formal training, the mean duration received was 2.8 days for drugs and 3.5 days for alcohol. OHPs reported that they did not feel sufficiently trained in this area. Most (65%) did not routinely include standardized screening tools or deliver 'brief interventions' (78%), although many reported that they would routinely ask about substance use when there was no clear indication of use (42%). The majority did not feel they were adequately supported (54%) in this work, nor did they have adequate resources for these patients within their organization (68%). CONCLUSIONS: OHPs see HCWs with substance-use problems as part of their work, but the support provided is likely limited by insufficient training and inadequate support.

The outward transport of cortisol by mammalian cells in vitro.

It has been determined that cortisol and a few other steroids are transported outward from certain mammalian cells growing in vitro. The extrusion process is temperature dependent, glucose dependent, saturable, and operates for only a few selected steroids. Many, but not all, steroids are able to block the extrusion process but are not themselves transported. The outward transport process for steroids has been found in mouse fibroblasts, mouse lymphoma cells, and functional mouse adrenal gland tumor cells growing in vitro. The transport process is not present in two varieties of cells cultured from human sources-HeLa or diploid fibroblasts, WI-38.

The product of the leu-3 cistron as a regulatory element for the production of the leucine biosynthetic enzymes of Neurospora.

A class of intracistronic (or closely linked) partial reversions of leu-3 mutations has been found to be conditionally constitutive with respect to the synthesis of isopropylmalate isomerase (specified by the leu-2 cistron) and beta-isopropylmalate dehydrogenase (specified by the leu-1 cistron), two of the enzymes of leucine biosynthesis in Neurospora. The intermediate level of enzyme production by these leu-3(cc) mutants is independent of the obligatory inducer effector, alpha-isopropylmalate, but dependent upon the presence of the branched-chain amino acids, isoleucine, valine and leucine. The properties of leu-3+, leu-3 and leu-3(cc) in heterokaryons indicate that the transnuclear regulatory activity of the leu-3 product varies specifically as a function of available effector molecules. The information presented suggests that the leu-3 cistron is responsible not only for the production of a "positive" regulatory substance necessary for the expression of the leu-1 and leu-2 cistrons, but that it probably serves also a coordinating role in the expression of many of the genes involved in branched-chain amino acid metabolism.

Change in chromosome number associated with a double deletion in the Neurospora crassa mitochondrial chromosome.

The mitochondrial genome of Neurospora is usually found in a single covalently closed circular 62-kbp DNA molecule. We report here that the mitochondrial genome of a phenotypic revertant of a stopper mutant (stp-ruv) is contained primarily in two separate, nonoverlapping, autonomously replicating circular chromosomes. The circles, one about 21 kbp and the other somewhat less than 36 kbp are derived from the most frequent classes of recombinant chromosomes (21 and 41 kbp) in the chromosomal population of mitochondria in the original stopper mutant. The new, more stable chromosomal configuration, is associated with the deletion of two sequences (1 kbp and 4 kbp) at the splice junctions of the two circles. The data suggest that both deletions are likely to have originated from a single recombinational event involved in generating the 36-kbp circle. Secondary, spontaneously arising derivatives of stp-ruv have been found to yield, at high copy number, short sections of the 21-kbp circle in covalently closed supercoiled circles varying from unit length to very high multimers. The amplified segments span a common segment likely to contain the replication origin of the 21-kbp chromosome.

Recombination and replication of plasmid-like derivatives of a short section of the mitochondrial chromosome of Neurospora crassa.

The 21-kbp mitochondrial chromosome of the stp-ruv strain of Neurospora crassa undergoes regional amplification yielding plasmid-like supercoiled circles varying in size from subunit length to very high multimers. A comparison of the base sequence of the five plasmids studied, with the region of the chromosome from which they were derived, indicated that the amplified chromosomal segments were determined by a recombination-excision process near or within two structurally distinctive regions. One of these, consisting of nearly uninterrupted strings of Cs and Gs straddling tandem PstI site direct repeats, could form an extended hairpin loop with only a few mismatches. It was found at or near the 5' exchange point of all of the plasmids. An extended 35-bp sequence containing 17-bp direct repeats was the primary 3' site of exchange. Base sequence changes were found in the vicinity of exchange points. Most notable of these was a G insertion and T to C transition within a section of the 5' region likely to form a hairpin loop, suggesting the involvement of a mismatch repair-like mechanism in the recombination process. The sequence, TATATAGACATATA, was identified as a likely candidate for the site of replication initiation. A nearly identical sequence was found common to all of the corresponding plasmids of Podospora anserina and was reported near the presumed replication origin of the Drosophila yakuba mitochondrial chromosome. A search of GenBank revealed a remarkable association of the consensus sequence, TATATAGAXATATA, with the plus strand of organelle DNA.

Phosphorylase kinase mediating the effects of cyclic AMP in muscle.

In the classic view of the control of phosphorylase b to a conversion by catecholamines, cyclic AMP acts as the second messenger stimulating the activity of cyclic AMP-dependent protein kinase to covalently modify phosphorylase kinase. Phosphorylation of phosphorylase kinase converts this enzyme form with a nonactivated to an activated form with a markedly higher activity at pH 7. There is now considerable evidence that the activity of phospphorylase kinase is also regulated by changeds in the Ca-2+ concentration. The activity of both nonactivated and activated phosphorylase kinase is stimulated by Ca-2+ in the range of concentrations that have been reported to occur in the sacroplasm of contracting muscle, with the activated pphosphorylase kinase having a lower K-alpha for Ca-2+. Thus there are at leaset two mechanisms for the regulation of phosphorylase kinase activity in muscle. These mechanisms may act independently or in concert in controlling glycogenolysis stimulated by catecholamines, anoxia, or tetanic electrical stimulation...

Role of the leu-3 cistron in the regulation of the synthesis of isoleucine and valine biosynthetic enzymes of Neurospora.

The production by Neurospora of the enzymes of isoleucine and valine synthesis in response to specific end product-derived signals depends upon the presence of an effective leu-3 regulatory product and its effector alpha-isopropylmalate (alpha-IPM). In leu-3(+) strains, threonine deaminase production is repressed as a function of available isoleucine, acetohydroxy acid synthetase as a function of valine, and the isomeroreductase and dihydroxy acid dehydratase as a function of isoleucine and leucine. In the absence of an effective leu-3 regulatory product, alpha-isopropylmalate, or both, the production of isoleucine and valine biosynthetic enzymes is fixed at or near fully repressed levels even under conditions of severe end product limitation. Thus, in addition to its involvement in the regulation of expression of the three structural genes of leucine synthesis, the leu-3 alpha-IPM regulatory product is necessary for full expression of at least four genes specifying the structure of the enzymes of isoleucine and valine synthesis. It is suggested that the leu-3 alpha-IPM regulatory element may facilitate transcription of the genetically dispersed cistrons either by imposing specificity on ribonucleic acid polymerase for structurally similar promoters adjacent to each of the cistrons or by "opening" promoters after interaction with nearly identical stretches of deoxyribonucleic acid near each of the structural genes.

The "golden section" and bias in perceptions of social consensus.

Meta-analytic examination of I28 false consensus effect issues supports the hypothesis that the "Golden Section" (61.8% group size) approximates the level of actual consensus that separates overestimation of consensus (group size < 61.8%) from underestimation (group size > 61.8%). Overestimation of the actual percentage of others who endorse one's own view increases as actual consensus decreases from 61.8%, and underestimation increases as it exceeds 61.8%. The form of the response (viz, a yes or no answer to a question) moderates this conclusion. The Golden Section holds for majorities and minorities defined by agreement with an issue. For majority and minority groups defined by disagreement, the inflection point is higher. Contrary to Mullen and Hu (1988), for agreeing majorities, the slope for consensus underestimation as a function of increased majority size does not differ from that of minority overestimation.

Ca++ inhibition of isoproterenol responses in mammalian skeletal muscle.

In noncontracting mouse hemidiaphragms incubated in modified Krebs-Ringer--bicarbonate buffer with 10 mM Ca++, isoproterenol-stimulated phosphorylase a formation, conversion of phosphorylase kinase to the activated form, elevation of cyclic AMP-dependent protein kinase activity ratios and increase in cyclic AMP concentrations were reduced 35 to 50% over the responses in buffer with 2.5 mM Ca++. In buffer with 10 mM Ca++, the initial rate of isoproterenol-stimulated cyclic AMP accumulation was 59% of that in buffer with 2.5 mM Ca++. The inhibitory action of Ca++ on cyclic AMP accumulation was antagonized by verapamil, but not by inhibitors of cyclic nucleotide phosphodiesterase activity. In buffer with 2.5 mM Ca++, isoproterenol-stimulated cyclic AMP accumulation was inhibited by A23187 and caffeine, agents that can increase intracellular Ca++ concentrations. In addition to Ca++, high concentrations of Co++, Ni++, Mn++ and, to a lesser extent, Sr++ inhibited the isoproterenol response. The results of these studies indicate that high buffer Ca++ concentrations inhibit the response of the glycogenolytic pathway to isoproterenol by an action on cyclic AMP formation. We propose that the site of the inhibitory action of Ca++ is the divalent metal activator site associated with hormone-stimulated adenylate cyclase activity.

Frequency modulation of the inotropic action of isoproterenol in mouse heart.

In mouse right ventricular strips, field-stimulated to contract isometrically in an oxygenated bicarbonate-buffered physiological salt solution at 22--24 degrees C, the EC50 for the inotropic action of isoproterenol decreased from 37 nM in muscles stimulated at 0.2 Hz to 5 nM in muscles stimulated at 3.3 Hz. At higher rates of contraction, there was also an increased sensitivity to the inotropic actions of norepinephrine and epinephrine but not to those of Ca++ and N6,O2'-dibutyryl cyclic AMP. Increasing the Ca++ concentration further decreased the EC50 for isoproterenol at 3.3 Hz but had no effect on the EC50 at 0.2 Hz. The leftward shift of the contractile response curve at 3.3 Hz was inhibited by verapamil (0.6 microM) and Mn++ (0.25 mM). The stimulation of cyclic AMP accumulation was approximately 6-fold more sensitive to isoproterenol at 3.3 than at 0.2 Hz, but isoproterenol increased contractile force at concentrations two orders of magnitude lower than those that significantly increased cyclic AMP accumulation. Inhibition of cyclic AMP phosphodiesterase activity further increased the sensitivity to the inotropic actions of isoproterenol but did not attenuate the frequency difference. The results indicate that isoproterenol-stimulated Ca++ influx through the slow channel plays an important role in the mechanism of the increased sensitivity to the inotropic action of isoproterenol found at higher frequencies of contraction. Although cyclic AMP accumulation was also frequency dependent, its role in the inotropic action of isoproterenol in mouse heart is not clear.

Rate dependence of isoproterenol-stimulated phosphorylase a formation in mouse heart.

In mouse right ventricular strips, field-stimulated to contract isometrically in an oxygenated bicarbonate-buffered physiological salt solution at 22--24 degrees C, isoproterenol (3 microM) stimulation of phosphorylase a accumulation was approximately a linear function of the rate of contraction from 0.2 to 5 Hz. In muscles incubated in the absence of isoproterenol, the phosphorylase a activity did not increase when the contraction frequency was varied over this range. At a low frequency of stimulation (0.2 Hz), phosphorylase a activity was not increased after a 5-min exposure to 3 microM isoproterenol, as compared to a 4-fold increase in phosphorylase a activity at a high frequency (3.-3 Hz). Isoproterenol (3 microM) increased tissue cyclic AMP content and the activated form of phosphorylase kinase activity to similar extents at both frequencies. N6,O2'-dibutyryl cyclic AMP increased the phosphorylase a activity at both frequencies but the increase at 3.3 Hz was approximately 3-fold greater than at 0.2 Hz. Verapamil did not block isoproterenol-stimulated phosphorylase a activity at 3.3 Hz at a concentration (0.6 microM) that inhibited the increased sensitivity to the inotropic action of isoproterenol seen at high frequencies of contraction. Isoproterenol stimulation of phosphorylase a accumulation did not correlate with developed tension. It is proposed that the difference in isoproterenol stimulation of phosphorylation b to a conversion at 0.2 and 3.3 Hz primarily results from a difference in Ca++ control of the activity of the activated form of phosphorylase kinase.

Purification of the three nuclear RNA polymerases from Neurospora crassa.

Nuclear RNA polymerases I, II, and III from Neurospora crassa have been purified 3,000-, 1,500-, and 10,000-fold, respectively, by a procedure that minimizes proteolysis of the 220-kDa subunit of polymerase II. The Neurospora enzymes resemble, in polypeptide composition, the corresponding polymerases isolated from other eukaryotes. The 220-kDa subunit of Neurospora polymerase II cross-reacts with antisera directed against the 220-kDa subunits of type II polymerases from Drosophila and wheat germ. However, the proteolyzed 180-kDa derivative of the Neurospora 220-kDa subunit fails to cross-react with the heterologous antisera, suggesting that the region removed by proteolysis contains or contributes to structural features of the enzyme that have been highly conserved during evolution. A 700-kDa complex of 12 polypeptides was found associated with polymerase II during purification. The complex was eventually separated from the enzyme, and its properties suggest that it might be associated with polymerase II in the nucleus. We describe two additional examples of polypeptides associated in variable amounts with Neurospora polymerase II.

Structural studies on Neurospora RNA polymerases and associated proteins.

Extensive structural homology between the three nuclear RNA polymerases of Neurospora crassa has been observed by peptide and immunological analyses. Within each polymerase, we found structural similarity between subunits in the 65- to 75-kDa range and one of the two large subunits. We observed also that polymerase II, as isolated, is associated with a 700-kDa complex of 12 polypeptides which is localized in the nucleus. A 75-kDa subunit of the 700-kDa complex was found to be structurally related to the 220-kDa subunit of polymerase II. We suggest, on the basis of the in vitro association, the common nuclear localization and the structural homology, that the 700-kDa complex and polymerase II may be associated in vivo. Evidence is also presented which suggests that polymerase III may interact with chromatin via two of its smallest subunits. A simple procedure for isolating nuclei from Neurospora is described.

Specific regulatory interconnection between the leucine and histidine pathways of Neurospora crassa.

Leucine auxotrophs of Neurospora fall into two discrete categories with respect to sensitivity to the herbicide, 3-amino-1,2,4-triazole. The pattern of resistance corresponds exactly to the ability to produce the leucine pathway control elements, alpha-isopropylmalate and the leu-3 product. An analysis of the regulatory response of the production of enzymes of histidine biosynthesis to alpha-isopropylmalate implicates the control elements of the leucine pathway as important components of the mechanism governing the production of the target enzyme of aminotriazole inhibition, imidazoleglycerol-phosphate dehydratase (EC 4.2.1.19). The evidence suggests that the regulatory interconnection between the two pathways is direct and is independent of other general integrating regulatory mechanisms which appear to be operative in both pathways. A general method for isolating leu-1 and leu-2, as well as other regulatory mutants, is described, which takes advantage of the specificity of the resistance to the inhibitor. Use of analogous systems is prescribed for the analysis of other regulatory interconnections which, like this one, might not be anticipated directly from structural or biosynthetic considerations.

Regulation of cytoplasmic and mitochondrial leucyl-transfer ribonucleic acid synthetases in Neurospora crassa.

The production of cytoplasmic leucyl-transfer ribonucleic acid synthetase activity was found to be reciprocally proportional to that of the corresponding mitochondrial enzyme during logarithmic growth of strains of Neurospora crassa with normal mitochondria. In the presence of cni-3 mutant mitochondria, production of mitochondrial enzyme activity was greatly increased whereas cytoplasmic enzyme production remained constant. The effect of cni-3 on the yield of the two enzyme activities indicated that the regulatory mechanism involved is a complicated one that cannot be accounted for by the relatively simple transcription competition model proposed previously.