Optimization of transgene action at the post-transcriptional level: high quality parthenocarpic fruits in industrial tomatoes

Background: Genetic engineering of parthenocarpy confers to horticultural plants the ability toproduce fruits under environmental conditions that curtail fruit productivity and quality. TheDefH9-iaaM transgene, whose predicted action is to confer auxin synthesis specifically in theplacenta, ovules and derived tissues, has been shown to confer parthenocarpy to several plantspecies (tobacco, eggplant, tomato) and varieties.Results: UC82 tomato plants, a typical cultivar used by the processing industry, transgenic for theDefH9-iaaM gene produce parthenocarpic fruits that are malformed. UC82 plants transgenic forthe DefH9-RI-iaaM, a DefH9-iaaM derivative gene modified in its 5'ULR by replacing 53 nucleotidesimmediately upstream of the AUG initiation codon with an 87 nucleotides-long sequence derivedfrom the rolA intron sequence, produce parthenocarpic fruits of high quality. In an in vitro translationsystem, the iaaM mRNA, modified in its 5'ULR is translated 3\u20134 times less efficiently than theoriginal transcript. An optimal expressivity of parthenocarpy correlates with a reduced transgenemRNA steady state level in DefH9-RI-iaaM flower buds in comparison to DefH9-iaaM flower buds.Consistent with the known function of the iaaM gene, flower buds transgenic for the DefH9-RI-iaaMgene contain ten times more IAA than control untransformed flower buds, but five times less thanDefH9-iaaM flower buds.Conclusions:: By using an auxin biosynthesis transgene downregulated at the post-transcriptionallevel, an optimal expressivity of parthenocarpy has been achieved in a genetic background notsuitable for the original transgene. Thus, the method allows the generation of a wider range ofexpressivity of the desired trait in transgenic plants

Plasma Retinol and Prognosis of Postmenopausal Breast Cancer Patients

Abstract Background: The role of retinol (vitamin A) in breast cancer prognosis has never been investigated in postmenopausal women. We prospectively assessed the long-term prognostic role of retinol plasma levels in a cohort of postmenopausal breast cancer patients. Patients and Methods: We investigated 208 women self-reported as postmenopausal operated on for T1-2N0M0 breast cancer who participated in a chemoprevention trial as controls and never received chemotherapy or hormone therapy. Plasma samples were collected 3 months (median) after surgery and assayed within 3 weeks for retinol. Minimum and median potential follow-up were 12 and 15 years, respectively. The main analyses were on all women and on a subgroup ages ≥55 years, assumed too old to be in perimenopause. The main end point was breast cancer death. Breast cancer survival was estimated by the Kaplan-Meier method. The hazard ratios of breast cancer death by retinol level were estimated by Cox models stratified for age, where relevant, and recruitment period, and adjusted for tumor size and histology. Results: At 12 years, patients with low retinol (<2.08 μmol/L, median of distribution) had lower breast cancer survival than those with high retinol (log-rank P = 0.052); the difference was significant for women ≥55 years (log-rank P = 0.006). The adjusted hazard ratios for low versus high retinol were 2.11 (95% confidence interval, 1.08-4.14) for all women and 3.58 (95% confidence interval, 1.50-8.57) for those ≥55 years. Conclusions: Low plasma retinol strongly predicts poorer prognosis in postmenopausal breast cancer patients. Retinol levels should be determined as part of the prognostic workup. (Cancer Epidemiol Biomarkers Prev 2009;18(1):42–8

Optimisation of transgene action at the post-transcriptional level: high quality parthenocarpic fruits in industrial tomatoes

Abstract Background Genetic engineering of parthenocarpy confers to horticultural plants the ability to produce fruits under environmental conditions that curtail fruit productivity and quality. The DefH9-iaaM transgene, whose predicted action is to confer auxin synthesis specifically in the placenta, ovules and derived tissues, has been shown to confer parthenocarpy to several plant species (tobacco, eggplant, tomato) and varieties. Results UC82 tomato plants, a typical cultivar used by the processing industry, transgenic for the DefH9-iaaM gene produce parthenocarpic fruits that are malformed. UC82 plants transgenic for the DefH9-RI-iaaM, a DefH9-iaaM derivative gene modified in its 5'ULR by replacing 53 nucleotides immediately upstream of the AUG initiation codon with an 87 nucleotides-long sequence derived from the rolA intron sequence, produce parthenocarpic fruits of high quality. In an in vitro translation system, the iaaM mRNA, modified in its 5'ULR is translated 3–4 times less efficiently than the original transcript. An optimal expressivity of parthenocarpy correlates with a reduced transgene mRNA steady state level in DefH9-RI-iaaM flower buds in comparison to DefH9-iaaM flower buds. Consistent with the known function of the iaaM gene, flower buds transgenic for the DefH9-RI-iaaM gene contain ten times more IAA than control untransformed flower buds, but five times less than DefH9-iaaM flower buds. Conclusions By using an auxin biosynthesis transgene downregulated at the post-transcriptional level, an optimal expressivity of parthenocarpy has been achieved in a genetic background not suitable for the original transgene. Thus, the method allows the generation of a wider range of expressivity of the desired trait in transgenic plants.</p

Yttrium-90 radioembolization for intermediate-advanced hepatocellular carcinoma: A phase 2 study

n/

Thiol-mediated protein retention in the endoplasmic reticulum: the role of ERp44

Formation of disulfide bonds, an essential step for the maturation and exit of secretory proteins from the endoplasmic reticulum (ER), is controlled by specific ER-resident enzymes. A pivotal element in this process is Ero1α, an oxidoreductin that lacks known ER retention motifs. Here we show that ERp44 mediates Ero1α ER localization through the formation of reversible mixed disulfides. ERp44 also prevents the secretion of an unassembled cargo protein with unpaired cysteines. We conclude that ERp44 is a key element in thiol-mediated retention. It might also favour the maturation of disulfide-linked oligomeric proteins and their quality control

Pulmonary nodules: volume repeatability at multidetector CT lung cancer screening.

To assess in vivo volumetric repeatability of an automated software algorithm in pulmonary nodules detected during a lung cancer screening trial.This study was approved by an institutional review board. Written informed consent was obtained from all participants. Data were collected from the Multicentric Italian Lung Detection project, a randomized controlled lung cancer screening trial. The first 1236 consecutive baseline computed tomographic (CT) studies performed at the Istituto Nazionale Tumori of Milan were evaluated. Among the enrolled participants, those who underwent repeat low-dose CT after 3 months and had at least one indeterminate nodule with a volume of more than 60 mm(3) (diameter of 4.8 mm or greater) were considered. Nonsolid, part-solid, and pleural-based nodules were excluded from this study. A descriptive analysis was performed by calculating means and standard deviations of nodule volumes at three assessment times (at baseline and 3 and 12 months later). The volume measurement repeatability was determined by using the approach described by Bland and Altman.One hundred one subjects (70 men, 31 women; mean age, 58 years) with 233 eligible nodules (mean volume, 98.3 mm(3); range, 5-869 mm(3)) were identified. The 95\% confidence interval for difference in measured volumes was in the range of +/-27\%. About 70\% of measurements had a relative difference in nodule volume of less than 10\%. No malignant lesions were registered during the follow-up of these subjects.Semiautomatic volumetry is sufficiently accurate and repeatable and may be useful in assisting with lung nodule management in a lung cancer screening program