DNA binding and transcription activation by chicken interferon regulatory factor-3 (chIRF-3).

Interferon regulatory factors (IRFs) are a family of transcription factors involved in the cellular response to interferons and viral infection. Previously we isolated an IRF from a chicken embryonic liver cDNA library. Using a PCR-based binding site selection assay, we have characterised the binding specificity of chIRF-3. The optimal binding site (OBS) fits within the consensus interferon-stimulated response element (ISRE) but the specificity of chIRF-3 binding allows less variation in nucleotides outside the core IRF-binding sequence. A comparison of IRF-1 and chIRF-3 binding to ISREs in electrophoretic mobility shift assays confirmed that the binding specificity of chIRF-3 was clearly distinguishable from IRF-1. The selection assay also showed that chIRF-3 is capable of binding an inverted repeat of two half OBSs separated by 10-13 nt. ChIRF-3 appears to bind both the OBS and inverted repeat sites as a dimer with the protein-protein interaction requiring a domain between amino acids 117 and 311. In transfection experiments expression of chIRF-3 strongly activated a promoter containing the OBS. The activation domain was mapped to between amino acids 138 and 221 and a domain inhibitory to activation was also mapped to the C-terminal portion of chIRF-3.

Cloning and promoter analysis of the chicken interferon regulatory factor-3 gene.

Interferon regulatory factors (IRFs) are a family of DNA-binding proteins involved in mediating the cellular response to interferons (IFNs) and viral infection. Although extensively studied in mammals, IRFs of other vertebrates have been less well characterized. Previously, we cloned chicken interferon regulatory factor-3 (chIRF-3) mRNA, which is rapidly and transiently induced by double-stranded (ds)RNA. The chIRF-3 mRNA encodes a protein distinct from any known mammalian IRF. Here, we show that chIRF-3 is activated additively by type I and type II IFNs. To delineate the sequence elements required to regulate chIRF-3 expression, we cloned chlRF-3 and 0.48 kb of 5' flanking sequence. Computer analysis of the proximal promoter revealed three putative binding sites for nuclear factor (NF)-kappaB, two overlapping interferon-stimulated response elements (ISREs), and an interferon gamma activating sequence (GAS). The presence of both GAS and ISRE consensus sequences in the chIRF-3 promoter is unique among IRF family members. Both type I and II IFNs, as well as dsRNA and IRF-1, trans-activate the promoter in short-term transfection experiments. Mutational analysis of the promoter demonstrated that the putative NF-kappaB binding sites are needed for stimulation by dsRNA but not by either type I or type II IFN and that both the overlapping ISREs and GAS are required for full induction by type I or type II IFN.

Establishment of interrater reliability for a nursing Extrapyramidal Side Effects (EPS) Assessment Scale.

Many patients who receive antipsychotic medications experience dystonia, akinesia, dyskinesia, and akathisia, collectively called Extrapyramidal Symptom Side Effects (EPS). The purpose of this study was to establish interrater reliability for a Nursing EPS Assessment Scale developed to focus on all four symptom areas. Twenty RNs and 12 patients participated in the instrument development studies. After several revisions, interrater reliability significance was demonstrated at the 0.01 level. It was concluded that the Nursing EPS Assessment Scale possessed good interrater reliability for nursing assessment of EPS.

The relationship between self-induced water intoxication and severity of psychiatric symptoms.

The purpose of this study was to test a theoretical framework that proposed a relationship between severe psychiatric symptoms and self-induced water intoxication (SIWI) by using reliable and valid measures. Twenty of 28 community-dwelling individuals with severe mental illness (SMI) in the sample exhibited excess fluid consumption as reflected by a mean urine specific gravity of 1.003 mEq/L. The psychometric measures included the Positive and Negative Syndrome Scale (PANSS), the Self Deficit Syndrome Scale (SDSS), and the Spielberger State-trait Anxiety Scale (STAI From X-1). A significant relationship was found at p .01 between severity of psychotic symptoms and severity of SIWI and between severity of SIWI and psychopathology symptoms within the PANSS. Anxiety was higher in those with SIWI before fluid loading compared with those with less excess fluid intake (i.e., USG < 1.010), and anxiety decreased from morning to evening in those with SIWI compared with those who did not exhibit excess fluid intake. The findings revealed a strong relationship between SIWI and severe psychiatric symptoms, including psychosis and a broad range of psychiatric symptoms. The findings provided initial support for the proposed theory, and consideration needs to be given to the development of interventions to augment existing treatment of fluid control.

Patient perceptions of self-induced water intoxication.

This study comprehensively examines self-induced water intoxication (SIWI) from a patient perspective including demographics, reasons for seeking fluids, patterns/behaviors of fluid seeking, and symptoms frequently experienced while in a state of SIWI. The subjects were 45 of an original convenience sample of 62 individuals with a serious mental illness (SMI), hospitalized in a long-term state psychiatric facility, who engaged in self-induced water intoxication. All participants were interviewed in a structured format to complete a 40-item Likert-type questionnaire developed for the study, titled the Self-induced Water Intoxication Questionnaire (SIWIQ). In the study, the majority of participants were smokers, and reported no past problem with alcohol. SIWI occurred more in males than females, and was more predominant in those participants who had longer hospital stays. Anger and vomiting were found to be the two most predominant symptoms experienced when excess fluid consumption occurred. Behaviors of drinking from the shower, the toilet, and one's own urine are consistent with findings of previous studies and illustrate the difficulty in keeping individuals with SIWI from fluids. Data show that participants with SIWI experience considerable anxiety and cognitive difficulties and express these as reasons for engaging in excess fluid consumption. Boredom, obtaining a high, and sad mood were also predominant reasons identified for excess fluid drinking. Significant relationships were found and discussed. The findings provide support for the position that SIWI represents an attempt at treatment by the dysfunctional individual and is pursued for anxiolytic effects and alleviation of boredom. The data support the idea of approaching the problem from a dysfunctional coping framework, realizing that SIWI is a very complex problem, needing examination and intervention at multiple levels, beyond exclusive focus on fluid control.

Isolation and characterization of the chicken homeodomain protein AKR.

Expression of the avian apoVLDLII gene is liver specific and completely dependent on estrogen. Previous analyses of protein binding sites in the apoVLDLII promoter revealed interactions between liver-enriched and ubiquitous factors at a location, site F', between nucleotides -229 and -260 relative to the major transcriptional start site. Site-directed mutagenesis of G residues contacted by these factors decreased expression from the promoter approximately 5-fold in the avian hepatoma cell line LMH2A. We have used this site to screen a cDNA expression library constructed from day 9 embryonic liver RNA. One of the two DNA binding factors isolated is a novel homeodomain protein. With the exception of the homeodomain itself, which is atypically located close to the protein N-terminus, the factor displays little similarity to any known DNA binding protein. Its homeodomain is most similar to that of the maize protein Knotted-1, while the most closely related vertebrate domain is that of the human proto-oncoprotein Pbx1. We demonstrate that the DNA binding specificity of the factor is consistent with its involvement in the ubiquitous complex formed with site F' and that it is capable of suppressing expression from the apoVLDLII promoter in short-term transfection experiments. These studies, combined with its DNA binding specificity, the tissue distribution of its mRNA and its developmental regulation, suggest a role as a negative regulator of gene expression in non-hepatic tissues and in the liver early during embryogenesis.

A comparison of clinical, morphological and immunohistochemical features of lymphomatoid papulosis and primary cutaneous CD30(Ki-1)-positive anaplastic large cell lymphoma.

The advent of immunoperoxidase technique on paraffin embedded tissue has identified a number of shared immunologic markers present in various lymphoproliferative cutaneous disorders. Two such disorders are the recently described primary cutaneous CD30-positive anaplastic large cell lymphoma (ALCL) and lymphomatoid papulosis; both entities are characterized by CD30-positive large atypical cells predominantly of T cell origin. We have compared the clinical, morphological and immunohistochemical features of 50 patients with lymphomatoid papulosis to a group of 27 patients with cutaneous CD30-positive ALCL. There are clear differences between the clinical presentation in these two diseases, and although both are characterized by similar atypical cells, the histologic pattern and distribution of atypical cells is sufficiently different to allow distinction and specific diagnosis based on hematoxylin and eosin stained sections supported by the immunohistochemical stains. In addition, both diseases are characterized by a long benign course, rarely complicated by development of lymphoreticular malignancy and invariably demonstrate CD30 (Ki-1) antigen positive large atypical cells.

Differential induction of cytolytic susceptibility by E1A, myc, and ras oncogenes in immortalized cells.

The E1A oncogene of adenovirus serotypes 2 and 5 induces susceptibility to the cytolytic effects of natural killer lymphocytes and activated macrophages when expressed in infected and transformed mammalian cells (cytolysis-susceptible phenotype). E1A and the oncogenes v-myc, long-terminal-repeat-promoted c-myc, and activated c-ras share the ability to immortalize transfected low-passage rodent cells. The cytolytic phenotypes of well-characterized rodent cell lines immortalized by these three oncogenes were defined. In contrast to target cells expressing the intact E1A gene, myc- and ras-expressing, immortalized primary transfectants were resistant to lysis by both types of killer cell populations. The same patterns of susceptibility (E1A) and resistance (myc and ras) to cytolysis were observed in oncogene-transfected continuous rat (REF52) and mouse (NIH 3T3) cell lines, indicating that differences in the cytolytic phenotypes associated with expression of these oncogenes are not due to cell selection during immortalization. The results suggest that the E1A oncogene may possess a functional domain that is different from those of other oncogenes, such as myc and ras, and that the activity linked to this postulated domain is dissociable from the process of immortalization.

Role of tumor necrosis factor-alpha in E1A oncogene-induced susceptibility of neoplastic cells to lysis by natural killer cells and activated macrophages.

NIH-3T3 cells transfected with adenovirus E1A oncogene cDNA were found to exhibit cytolytic susceptibility to murine NK cells and activated macrophages associated with a threshold level of oncogene product expression exceeding that required for morphological transformation. A similar correlation was observed between threshold levels of E1A gene product expression and target cell susceptibility to direct cytotoxicity by rTNF. Inhibition of splenic NK cell and peritoneal macrophage cytolysis by antisera specific for murine rTNF confirmed the importance of E1A-induced TNF susceptibility as one determinant of target cell cytolytic susceptibility. Anti-TNF antibody was, however, unable to block killing of E1A-expressing targets by the NK cell line, NKB61A2. These results suggest a direct link between the functions of E1A oncogene products and cellular mechanisms of action of TNF elaborated by host effector cells and indicate that E1A expression also affects target cell susceptibility to TNF-independent cytolytic mechanisms.

Adenovirus E1A gene induction of susceptibility to lysis by natural killer cells and activated macrophages in infected rodent cells.

Rodent cells immortalized by the E1A gene of nononcogenic adenoviruses are susceptible to lysis by natural killer (NK) cells and activated macrophages. This cytolysis-susceptible phenotype may contribute to the rejection of adenovirus-transformed cells by immunocompetent animals. Such increased cytolytic susceptibility has also been observed with infected rodent cells. This infection model provided a means to study the role of E1A gene products in induction of cytolytic susceptibility without cell selection during transformation. Deletion mutations outside of the E1A gene had no effect on adenovirus type 2 (Ad2) or Ad5 induction of cytolytic susceptibility in infected hamster cells, while E1A-minus mutant viruses could not induce this phenotype. E1A mutant viruses that induced expression of either E1A 12S or 13S mRNA in infected cells were competent to induce cytolytic susceptibility. Furthermore, there was a correlation between the accumulation of E1A gene products in Ad5-infected cells and the level of susceptibility of such target cells to lysis by NK cells. The results of coinfection studies indicated that the E1A gene products of highly oncogenic Ad12 could not complement the lack of induction of cytolytic susceptibility by E1A-minus Ad5 virus in infected cells and also could not block induction of this infected-cell phenotype by Ad5. These data suggest that expression of the E1A gene of nononcogenic adenoviruses may cause the elimination of infected cells by the immunologically nonspecific host inflammatory cell response prior to cellular transformation. The lack of induction of this cytolysis-susceptible phenotype by Ad12 E1A may result in an increased persistence of Ad12-infected cells in vivo and may lead to an increased Ad12-transformed cell burden for the host.

Treatment of multiple keratoacanthomas with intralesional fluorouracil.

A keratoacanthoma is a benign epithelial growth most often found in sun-exposed skin of the elderly. Two main groups of multiple keratoacanthomas have been described: the Ferguson-Smith type and the Grzybowski type. A patient with multiple keratoacanthomas of the Ferguson Smith type is described and the treatment with intralesional 5-fluorouracil is explained. Intralesional therapy with 5-fluorouracil is a safe treatment and may be the treatment of choice for multiple keratoacanthomas and solitary lesions which are difficult to remove surgically.