Electrophilic nitro-fatty acids suppress allergic contact dermatitis in mice.

BACKGROUND: Reactions between nitric oxide (NO), nitrite (NO2-), and unsaturated fatty acids give rise to electrophilic nitro-fatty acids (NO2 -FAs), such as nitro oleic acid (OA-NO2 ) and nitro linoleic acid (LNO2 ). Endogenous electrophilic fatty acids (EFAs) mediate anti-inflammatory responses by modulating metabolic and inflammatory signal transduction reactions. Hence, there is considerable interest in employing NO2 -FAs and other EFAs for the prevention and treatment of inflammatory disorders. Thus, we sought to determine whether OA-NO2 , an exemplary nitro-fatty acid, has the capacity to inhibit cutaneous inflammation. METHODS: We evaluated the effect of OA-NO2 on allergic contact dermatitis (ACD) using an established model of contact hypersensitivity in C57Bl/6 mice utilizing 2,4-dinitrofluorobenzene as the hapten. RESULTS: We found that subcutaneous (SC) OA-NO2 injections administered 18 h prior to sensitization and elicitation suppresses ACD in both preventative and therapeutic models. In vivo SC OA-NO2 significantly inhibits pathways that lead to inflammatory cell infiltration and the production of inflammatory cytokines in the skin. Moreover, OA-NO2 is capable of enhancing regulatory T-cell activity. Thus, OA-NO2 treatment results in anti-inflammatory effects capable of inhibiting ACD by inducing immunosuppressive responses. CONCLUSION: Overall, these results support the development of OA-NO2 as a promising therapeutic for ACD and provides new insights into the role of electrophilic fatty acids in the control of cutaneous immune responses potentially relevant to a broad range of allergic and inflammatory skin diseases.

An audit of smoking prevalence and awareness of HSE smoking cessation services among HSE staff.

This audit estimated smoking prevalence and awareness of quit services among Health Service Executive (HSE) staff. A questionnaire posted to a random sample of 1,064 staff received a 71% response rate. Staff smoking prevalence was 15.0% overall, and 4.4% among Medical/Dental staff. Front-line-healthcare staff were less likely to smoke than other staff categories (adjusted OR 0.38, p < 0.001). Only 63.6% of staff were aware of HSE quit services. Targeted interventions are required to help staff to quit smoking and to boost awareness of quit services.

The general transcription factor RAP30 binds to RNA polymerase II and prevents it from binding nonspecifically to DNA.

RAP30/74 is a human general transcription factor that binds to RNA polymerase II and is required for initiation of transcription in vitro regardless of whether the promoter has a recognizable TATA box (Z. F. Burton, M. Killeen, M. Sopta, L. G. Ortolan, and J. F. Greenblatt, Mol. Cell. Biol. 8:1602-1613, 1988). Part of the amino acid sequence of RAP30, the small subunit of RAP30/74, has limited homology with part of Escherichia coli sigma 70 (M. Sopta, Z. F. Burton, and J. Greenblatt, Nature (London) 341:410-414, 1989). To determine which sigmalike activities of RAP30/74 could be attributed to RAP30, we purified human RAP30 and a RAP30-glutathione-S-transferase fusion protein that had been produced in E. coli. Bacterially produced RAP30 bound to RNA polymerase II in the absence of RAP74. Both partially purified natural RAP30/74 and recombinant RAP30 prevented RNA polymerase II from binding nonspecifically to DNA. In addition, nonspecific transcription by RNA polymerase II was greatly inhibited by RAP30-glutathione-S-transferase. DNA-bound RNA polymerase II could be removed from DNA by partially purified RAP30/74 but not by bacterially expressed RAP30. Thus, the ability of RAP30/74 to recruit RNA polymerase II to a promoter-bound preinitiation complex may be an indirect consequence of its ability to suppress nonspecific binding of RNA polymerase II to DNA.

RAP30/74: a general initiation factor that binds to RNA polymerase II.

We have previously shown by affinity chromatography that RAP30 and RAP74 are the mammalian proteins that have the highest affinity for RNA polymerase II. Here we show that RAP30 binds to RAP74 and that the RAP30-RAP74 complex (RAP30/74) is required for accurate initiation by RNA polymerase II. RAP30/74 is required for accurate transcription from the following promoters: the adenovirus major late promoter, the long terminal repeat of human immunodeficiency virus, P2 of the human c-myc gene, the mouse beta maj-globin promoter (all of which have TATA boxes), and the mouse dihydrofolate reductase promoter (which lacks a TATA box). RAP30/74 is not required for initiation by RNA polymerase III at the adenovirus virus-associated RNA promoters. Therefore, RAP30/74 is a general initiation factor that binds to RNA polymerase II.

Multiple signaling mechanisms of the UNC-6/netrin receptors UNC-5 and UNC-40/DCC in vivo.

Cell and growth cone migrations along the dorsoventral axis of Caenorhabditis elegans are mediated by the UNC-5 and UNC-40 receptor subtypes for the secreted UNC-6 guidance cue. To characterize UNC-6 receptor function in vivo, we have examined genetic interactions between unc-5 and unc-40 in the migrations of the hermaphrodite distal tip cells. We report that cell migration defects as severe as those associated with a null mutation in unc-6 are produced only by null mutations in both unc-5 and unc-40, indicating that either receptor retains some partial function in the absence of the other. We show that hypomorphic unc-5 alleles exhibit two distinct types of interallelic genetic interactions. In an unc-40 wild-type genetic background, some pairs of hypomorphic unc-5 alleles exhibit a partial allelic complementation. In an unc-40 null background, however, we observed that unc-5 hypomorphs exhibit dominant negative effects. We propose that the UNC-5 and UNC-40 netrin receptors can function to mediate chemorepulsion in DTC migrations either independently or together, and the observed genetic interactions suggest that this flexibility in modes of signaling results from the formation of a variety of oligomeric receptor complexes.

Identification of three mammalian proteins that bind to the yeast TATA box protein TFIID.

The TATA box binding transcription factor TFIID of S. cerevisiae was used as a ligand for affinity chromatography. Polypeptides that bind specifically to yeast TFIID (TFIID-associated proteins, DAPs) were purified from human HeLa (heDAPs) and calf thymus (ctDAPs) whole cell extracts. Both heDAP and ctDAP fractions altered the binding of TFIID to the TATA element, and substituted for the TFIIA transcription activity in a reconstituted in vitro system. The heDAP fraction also behaved like TFIIA in its ability to form a promoter-TFIID-TFIIA complex and to recruit TFIIB to such a complex. The interaction of DAPs with TFIID can confer heat-resistance (47 degrees C) on recombinant yeast or human TFIID. SDS-PAGE analysis revealed that three polypeptides from HeLa extracts specifically bound to yTFIID columns (heDAP35, heDAP21, and heDAP12). These data suggest that a multi-subunit transcription factor with the properties of TFIIA can bind to TFIID in the absence of DNA.

Transbilayer migration (flip-flop) of 12-(4-azido-2-nitrophenoxy)stearoyl glucosamine in large unilamellar phospholipid vesicles.

The ability of the glycolipid photoprobe, 12-(4-azido-2-nitrophenoxy)-stearoyl[1-14C]glucosamine (12-APS-GlcN), to undergo transbilayer flip-flop and intermembrane transfer between liposomes was examined. It was found that probe which was incorporated into membranes during the preparation of large unilamellar vesicles (LUVs) could be rapidly and completely extracted by incubation of these donor vesicles (in the liquid-crystalline state) with probe-free acceptor vesicles.

Calibration of a horizontal pendulum-type tablet breaking strength tester.

A rapid, simplified method of checking the calibration of a new motorized pendulum type of tablet hardness tester and the accuracy and precision of three testers were determined. Two force gauges were calibrated, and both gauges were used by two operators to determine the accuracy of the new testers. The calibration method using the force gauge was reliable for checking the accuracy. The results show the various new testers to be in calibration and to give reproducible results.

Impact of case management on hospital charges of PWAs in Georgia.

To determine the effects of case management in controlling hospital based costs for persons with AIDS (PWAs), a retrospective review of records of deceased PWAs was performed. The study compared diagnosis-to-death hospital charges for PWAs who received care under a care management model (n = 60) with PWAs receiving care within a non-case managed approach (n = 60). Hospital charges were adjusted for inflation. PWAs receiving care within the case managed model had significantly lower hospital-based changes than the non-case managed group. Additionally, PWAs in the case managed group lived significantly longer between HIV diagnosis and death, and lived longer between their first AIDS-related hospital admission and death.

Getting through our grief. For caregivers of persons with AIDS.

Staff caring for AIDS patients experience a grief characterized by specific features. This article presents ways to understand these losses and develop supportive ways to manage their reactions.

Nursing fundamentals texts: where's the ethics?

A primary source of information on professional ethics for nursing students is the nursing fundamentals text used in the initial courses. This study systematically evaluated nursing fundamentals texts for their coverage of ethically relevant content. Forty-two nursing fundamentals texts published from 1965 to 1985 were evaluated for: inclusion of a professional code of ethics; interpretive statements, discussion and examples; and guidelines for ethical decision making. Forty-five percent of the texts contained no content on ethics, with the remainder varying in their depth of coverage. Analysis revealed a moderate correlation (r = 0.59) between the year of publication and the average number of pages accorded content on ethics in that year. The results reflect a gradual, although irregular, trend toward inclusion of greater amounts of content on ethics. Recommendations were proposed that would facilitate the inclusion of content on ethics into nursing curricula. These included: greater communication between faculty and text publishers regarding the importance of content on ethics, inclusion of ethics as a curricular thread, small-group workshops to increase faculty sensitivity to the need for preparation in ethical decision making, and an elective course in nursing ethics.

Maintenance of self-esteem by obese children.

Forty-five fifth grade children participated in a study to examine whether obese children differ in self-esteem from their average-weight peers. The study also explored whether obese children use the processes of discounting and cognitive distortion to maintain self-esteem. The obese children had lower scores than average-weight children in global self-worth, and in perceived competence in physical appearance and social acceptance, but not in scholastic competence, athletic competence, or behavioral conduct. The pattern of results indicate that obese children do not discount the importance of physical appearance and weight, but use distortion. Further, the use of distortion by obese children was associated with more positive self-perceptions. Although the use of defensive cognitions was effective in enhancing obese children's global self-worth, it was not powerful enough to prevent completely the negative effects of obesity on self-esteem.

Home care A to Z.

My home care agency gives out little gold pins to employees to mark an anniversary. Now that I have one I feel confident in telling the real story about home care. I don't mean the dull paragraphs buried in the Policies and Procedures Manual or the dry information presented in our agency's mission. I am talking about the truth that every social worker, nurse, home care aide, or physical or occupational or speech therapist faces daily in visits to clients' homes. I dream of the day that this primer will be integrated into each new staff person's orientation. Here is the working bible of home care from A to Z.

Acute atrial arrhythmogenesis in murine hearts following enhanced extracellular Ca(2+) entry depends on intracellular Ca(2+) stores.

AIM: To investigate the effect of increases in extracellular Ca(2+) entry produced by the L-type Ca(2+) channel agonist FPL-64176 (FPL) upon acute atrial arrhythmogenesis in intact Langendorff-perfused mouse hearts and its dependence upon diastolic Ca(2+) release from sarcoplasmic reticular Ca(2+) stores. METHODS: Confocal microscope studies of Fluo-3 fluorescence in isolated atrial myocytes were performed in parallel with electrophysiological examination of Langendorff-perfused mouse hearts. RESULTS: Atrial myocytes stimulated at 1 Hz and exposed to FPL (0.1 microm) initially showed (<10 min) frequent, often multiple, diastolic peaks following the evoked Ca(2+) transients whose amplitudes remained close to control values. With continued pacing (>10 min) this reverted to a regular pattern of evoked transients with increased amplitudes but in which diastolic peaks were absent. Higher FPL concentrations (1.0 microm) produced sustained and irregular patterns of cytosolic Ca(2+) activity, independent of pacing. Nifedipine (0.5 microm), and caffeine (1.0 mm) and cyclopiazonic acid (CPA) (0.15 microm) pre-treatments respectively produced immediate and gradual reductions in the F/F(0) peaks. Such nifedipine and caffeine, or CPA pre-treatments, abolished, or reduced, the effects of 0.1 and 1.0 microm FPL on cytosolic Ca(2+) signals. FPL (1.0 microm) increased the incidence of atrial tachycardia and fibrillation in intact Langendorff-perfused hearts without altering atrial effective refractory periods. These effects were inhibited by nifedipine and caffeine, and reduced by CPA. CONCLUSION: Enhanced extracellular Ca(2+) entry exerts acute atrial arrhythmogenic effects that is nevertheless dependent upon diastolic Ca(2+) release. These findings complement reports that associate established, chronic, atrial arrhythmogenesis with decreased overall inward Ca(2+) current.