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Background and objective: The aim of the Pulmonary Rehabilitation Guidelines (Guidelines) is to provide evidence-based recommendations for the practice of pulmonary rehabilitation (PR) specific to Australian and New Zealand healthcare contexts.

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Plasmid vectors remain a valuable yet capricious tool for the genetic manipulation of human embryonic stem (hES) cells. We have compared the efficacy of four promoters to mediate transient and stable transfection in hES and human embryonal carcinoma cell lines with the reporter enhanced green fluorescent protein (eGFP). In transient assays, the two mammalian promoters, UbiquitinC and Rosa26 (pUbiC and pR26), the human cytomegalovirus major immediate early promoter (HCMV-MIE; pCMV), and the HCMV-MIE/ chicken ␤-actin/rabbit ␤-globin hybrid promoter (pCAGG) gave variable results that depended upon the cell line transfected but in an unpredictable way: each promoter supported strong transient expression in at least one cell line. The results for stable transfection were generally at variance with the transient assays. In each case, transgene silencing was quite marked, most notably with the pCMV, with which no eGFP-positive clones were obtained. An exception was the pCAG vector, in which the CAGG composite promoter is linked to the polyoma virus mutant enhancer PyF101; stable eGFP-positive transfectants were obtained, and these clones retained eGFP expression for over 120 passages, even in the absence of selection. However, if the PyF101 elements were removed, the resulting transfectants were also subjected to progressive gene silencing. Thus, the choice of promoter is critical for determining the desired effect of transgene expression in hES cells. Our data also demonstrate that pUbiC, pR26, pCAGG, and pCAG are more superior to the pCMV for generation of stable transfectants in hES cells. STEM CELLS 2007;25:1521-1528 Disclosure of potential conflicts of interest is found at the end of this article.

Embryonal carcinoma (EC) cells, the pluripotent stem cells of teratocarcinomas, show many similar-ities to embryonic stem (ES) cells. Since EC cells are malignant but their terminally differentiated derivatives are not, understanding the molecular mechanisms that regulate their differentiation maybe of value for diagnostic and therapeutic purposes. We have examined the expression of multiple components of two developmentally important cell-cell signalling pathways, Wnt and Notch, in the pluripotent human EC cell line, NTERA2, and the human ES cell line, H7. Both pathways have well-documented roles in controlling neurogenesis, a process that occurs largely in response to retinoicacid (RA) treatment of NTERA2 cultures and spontaneously in H7 cultures. In NTERA2, many ofthe genes tested showed altered transcriptional regulation following treatment with RA. These include members of the frizzled gene family (FZDI, FZD3, FZD4, FZD5, FZD6), encoding receptors forWnt proteins, the Frizzled Related Protein family (SFRPI, SFRP2, FRZB, SFRP4), encoding solubleWnt antagonists and also ligands and receptors of the Notch pathway (Dlkl, Jaggedl; Notchl, Notch2, Notch3). Few differences were found in the repertoire of Wnt and Notch pathway genes expressed by NTERA2 EC cells and H7 ES cells. We present a model in which interactions between and regulation of Wnt and Notch signalling are important in maintaining EC/ES stem cells and also controlling their differentiation.

Background: The study aimed to determine the effects of adding cognitive behavioural therapy (CBT) to pulmonary rehabilitation to treat patients with chronic lung disease and comorbid anxiety and/or depression symptoms.Methods: An open, parallel group, randomised controlled trial (RCT) was conducted, with longitudinal follow-up of 12 months. CBT was delivered in 2 face-to-face sessions and 4 phone sessions to patients with depression or anxiety undergoing pulmonary rehabilitation. The main outcome measures were change in Geriatric Depression Scale (GDS) and Geriatric Anxiety Inventory (GAI); secondary outcomes were St.Georges Respiratory Questionnaire (SGRQ), 6-minute walk test (6MWT) and pulmonary rehabilitation attendance.Results: A total of 65 patients were randomized to Intervention (n=24) and Control (n=41) groups. Of the 24 patients in the Intervention group, 6 patients (25%) withdrew and 4 patients (12.5%) failed to attend more than 2 CBT sessions, which was significantly more than the Control group. The majority of patients (75.4%) had chronic obstructive pulmonary disease. Fourteen (21.5%) had symptoms of depression only, 12 (18.4%) had symptoms of anxiety only, and 39 (60.0%) had symptoms of both anxiety and depression. In the Intervention group, GDS significantly improved at the end of pulmonary rehabilitation (mean difference −3.1, 95% CI: −4.39 to −1.70; P=0.0001), 3 months follow-up (mean difference −1.5, 95% CI: −4.17 to −0.75; P=0.008), and at 12 months follow-up (mean difference −1.6, 95% CI: −3.29 to −0.03, P=0.04), compared to baseline. The Control group demonstrated improvement in GDS by the end of pulmonary rehabilitation (mean difference −1.3, 95% CI: −2.4 to −0.27; P=0.01) which was not maintained at 3 months (P=0.14) and 12 months (P=0.25). GAI significantly improved by the end of rehabilitation in both the Intervention (mean difference −2.6, 95% −4.69 to −0.57; P=0.01) and Control groups (mean difference −2.6, 95% −4.16 to −1.14; P=0.001) and there was no significant improvement at 3 and 12 months. No statistically significant differences in changes in GDS or GAI were observed between the Intervention and Control groups at any time point. There was no significant improvement in SGRQ or 6MWT. There was a significant increase in attended pulmonary rehabilitation sessions in the Intervention group, compared to the Control group (mean difference 1.59; 95% CI: 0.11 to 3.07; P=0.03). Conclusions: In this RCT of patients with chronic lung diseases attending pulmonary rehabilitation, there was no evidence found for improved symptoms of anxiety or depression or health-related quality of life with the addition of CBT given in a mixed face-to-face and telephone format, compared to usual care. Slower