Transcription of the antiatherogenic protein apolipoprotein AI is regulated by the thyroid hormone, L-triiodothyronine. Transient transfection and electrophoretic mobility shift assays were used to identify the cis-acting elements involved. In transient transfection assays, hormone bound to either thyroid hormone receptor alpha or beta exerts a positive effect through a thyroid hormone response element, site A (-208 to -193). In the absence of site A, liganded receptor alpha or beta have a negative effect on promoter activity. This negative effect is mediated by a 40 bp fragment spanning nucleotides -46 to -7. Closer examination of this region of the gene shows there to be a negative thyroid hormone response element at position -25 to -20 which is fused to the 3' end of the TATA element. Electrophoretic mobility shift assays show that bacterially expressed chicken or rat thyroid hormone receptor alpha 1 binds to site A, either as a homodimer or as a heterodimer with the human 9-cis-retinoic acid receptor alpha. In contrast, the negative thyroid hormone responsive element binds chicken thyroid hormone receptor alpha exclusively as a monomer. Site-directed mutagenesis of the negative thyroid hormone response element abolished the inhibitory effects of the hormone and increased basal promoter activity by up to 40-fold. These data suggest that functional positive and negative thyroid hormone response elements coexist within the rat apolipoprotein AI promoter and both elements contribute to the control of apolipoprotein AI gene expression.

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Little is known about the process of how nurses transition between vocational training and institutions of higher education. Understanding this process provides educators with the knowledge to support new groups of university students making this transition. Grounded theory (GT) was used to explore and understand this process. Three studies from a 7-year research program were used as data. The analysis led to the generation of a GT illuminating the process of students transitioning from post-LPN to BN. This GT illustrates how students overcome difficulties encountered moving to a more complex nursing role. The students' main concern was a lack of independence. The core variable, which resolves this main concern, and which emerged from the analysis of the data is developing independence. There are three sub-core variables, resisting, reaching out and re-imagining which support this core variable of developing independence.

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A chemically synthesized gene for human interferon-y has been cloned into a prokaryotic expression vector under the regulation of a synthetic constitutive transcriptional-translational control unit that contains a strong bacteriophage T5 early promoter and a strong ribosome-binding site. Cells harboring the recombinant plasmid express high levels (4 x 109 units per liter of culture) of antiviral activity specific for interferon-y. Analysis of total cell lysates on NaDodSO4/polyacrylamide gels revealed a 17,200-dalton protein, expected for the nonglycosylated form of human interferon-y, that constitutes >15% of total cell protein.Human interferon-y (HuIFN--y) is an interesting protein that exhibits a number of different biological activities. In addition to its antiviral activity, HuIFN-y has been shown to exhibit potent immunomodulation and cell proliferation-inhibition properties (reviewed in ref. 1). Recently, the nucleotide sequence of its gene has been determined (2, 3), and its expression in Escherichia coli has been achieved by using natural control signals from either the trp operon (2) or the lac UV5 operon (4). However, the reported yields were relatively low compared to those for IFN-a (5-7) and IFN-,B (8, 9).Among the large numbers of DNA fragments containing bacterial promoters that have been used for in vitro binding studies, the bacteriophage T5 early promoters (T5P25 and T5P26) have been found to far exceed other promoter fragments in the rate of complex formation with E. coli RNA polymerase (10, 11). Recently, we have reported the chemical synthesis and insertion of the T5P25 promoter in front of either the tetracycline-resistance (TcR) gene or the chloramphenicol acetyltransferase (CAT) gene, and have demonstrated that the synthetic T5P25 promoter is highly efficient in vivo (12).We have also shown that the introduction of a synthetic ribosome-binding site (RBS) in front of the coding sequence for simian virus 40 small tumor antigen, and its subsequent insertion at the Pst I site within the pBR322 ampicillin-resistance (ApR) gene, resulted in the synthesis of authentic simian virus 40 small tumor antigen in E. coli (13). Using this model system, we were able to compare the efficiency of different synthetic RBS sequences in vivo (14).In this communication, we describe the construction of a plasmid expression vector containing both the synthetic T5P25 promoter and a strong synthetic RBS; this vector (pJPlR3) is used for the efficient expression of a synthetic gene for human IFN-y. MATERIALS AND METHODSConstruction of the IFN-y Gene. The deoxyoligonucleotides (Fig. 1) comprising the entire IFN-y sequence including initiation and termination signals were chemically synthesized using a modified solid-phase phosphite method. -Details of the synthesis and construction of the gene will be published elsewhere.Construction of the pJPjR3 Expression Vector. The pJPj plasmid DNA, a derivative of pBR322 in which the TcI promoter between the EcoRI and HindIII sites has been replaced by the strong synth...

This study used Grounded Theory methodology and developed an emergent theory of Mutual Caring. The main concern was Habituation to ineffective lessons learned sharing practices. Habituation is resolved through Mutual Caring, a socially and psychologically adaptive process. Mutual Caring involves comfortable conversations, engaging/sharing, and developing self‐confidence, resulting in an enhanced wisdom pool. This paper extends the emphasis on tangible knowledge‐sharing mechanisms. Mutual Caring could lead to better outcomes, such as more accessible and usable knowledge, a project management wisdom pool, and criteria for improved dyadic relationships to enhance project learning.

BackgroundThe Self-Perception and Relationships Tool (S-PRT) is intended to be a clinically responsive and holistic assessment of patients' experience of illness and subjective Health Related Quality of Life (HRQL).MethodsA diversity of patients were involved in two phases of this study. Patient samples included individuals involved with renal, cardiology, psychiatric, cancer, chronic pelvic pain, and sleep services. In Phase I, five patient focus groups generated 128 perceptual rating scales. These scales described important characteristics of illness-related experience within six life domains (i.e., Physical, Mental-Emotional, Interpersonal Receptiveness, Interpersonal Contribution, Transpersonal Receptiveness and Transpersonal Orientation). Item reduction was accomplished using Importance Q-sort and Importance Checklist methodologies with 150 patients across the participating services. In Phase II, a refined item pool (88 items) was administered along with measures of health status (SF-36) and spiritual beliefs (Spiritual Involvements and Beliefs Scale – SIBS) to 160 patients, of these 136 patients returned complete response sets.ResultsFactor analysis of S-PRT results produced a surprisingly clean five-factor solution (Eigen values> 2.0 explaining 73.5% of the pooled variance). Items with weaker or split loadings were removed leaving 36 items to form the final S-PRT rating scales; Intrapersonal Well-being (physical, mental & emotional items), Interpersonal Receptivity, Interpersonal Contribution, Transpersonal Receptivity and Transpersonal Orientation (Eigen values> 5.4 explaining 83.5% of the pooled variance). The internal consistency (Cronbach's Alpha) of these scales was very high (0.82–0.97). Good convergent correlations (0.40 to 0.67) were observed between the S-PRT scales and the Mental Health scales of the SF-36. Correlations between the S-PRT Intrapersonal Well-being scale and three of SF-36 Physical Health scales were moderate (0.30 to 0.46). The criterion-related validity of the S-PRT spiritual scales was supported by moderate convergence (0.40–0.49) with three SIBS scales.ConclusionEvidence supports the validity of the S-PRT as a generally applicable measure of perceived health status and HRQL. The test-retest reliability was found to be adequate for most scales, and there is some preliminary evidence that the S-PRT is responsive to patient-reported changes in determinants of their HRQL. Clinical uses and directions for future research are discussed.

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