Quantification of the magnification and distortion effects of a pediatric flexible video-bronchoscope

Background

Flexible video bronchoscopes, in particular the Olympus BF Type 3C160, are commonly used in pediatric respiratory medicine. There is no data on the magnification and distortion effects of these bronchoscopes yet important clinical decisions are made from the images. The aim of this study was to systematically describe the magnification and distortion of flexible bronchoscope images taken at various distances from the object.

Methods

Using images of known objects and processing these by digital video and computer programs both magnification and distortion scales were derived.

Results

Magnification changes as a linear function between 100 mm (×1) and 10 mm (×9.55) and then as an exponential function between 10 mm and 3 mm (×40) from the object. Magnification depends on the axis of orientation of the object to the optic axis or geometrical axis of the bronchoscope. Magnification also varies across the field of view with the central magnification being 39% greater than at the periphery of the field of view at 15 mm from the object. However, in the paediatric situation the diameter of the orifices is usually less than 10 mm and thus this limits the exposure to these peripheral limits of magnification reduction. Intraclass correlations for measurements and repeatability studies between instruments are very high, r = 0.96. Distortion occurs as both barrel and geometric types but both types are heterogeneous across the field of view. Distortion of geometric type ranges up to 30% at 3 mm from the object but may be as low as 5% depending on the position of the object in relation to the optic axis.

Conclusion

We conclude that the optimal working distance range is between 40 and 10 mm from the object. However the clinician should be cognisant of both variations in magnification and distortion in clinical judgements.     

Agronomic biofortification of Brassica with selenium—enrichment of SeMet and its identification in Brassica seeds and meal

Selenium (Se) is an essential micronutrient and is circulated to the food chain through crops. Brassica species are efficient in Se accumulation and thus, good species for Se biofortification purposes. The residual fraction obtained after oil processing of Brassica seeds, the meal, is an important protein source in animal diets and used in feed concentrates. The accumulation of soil or foliar applied Se in the seeds and meal of Brassica napus and B. rapa as well as its effects on growth and yield formation was studied in two field experiments. Also, a HPLC-ICP-MS based method for the identification and quantification of Se species in Brassica seeds and meal was developed. Selenium application did not affect the yield or oil content. High accumulation of Se in the seeds and meal (1.92–1.96 μg Se g^?1) was detected. Biotransformation of inorganic Se was evaluated by using HPLC-ICP-MS previous enzymatic hydrolysis for species extraction. The Se speciation studies showed that up to 85% of the total Se was SeMet whereas other Se-species were not detected. We conclude that the agronomic biofortification of Brassica species can improve the nutritive quality of the protein rich meal fraction as it contains significant amount of SeMet.     

A Panel of Stably Expressed Reference Genes for Real-Time qPCR Gene Expression Studies of Mallards (Anas platyrhynchos)

Determining which reference genes have the highest stability, and are therefore appropriate for normalising data, is a crucial step in the design of real-time quantitative PCR (qPCR) gene expression studies. This is particularly warranted in non-model and ecologically important species for which appropriate reference genes are lacking, such as the mallard—a key reservoir of many diseases with relevance for human and livestock health. Previous studies assessing gene expression changes as a consequence of infection in mallards have nearly universally used β-actin and/or GAPDH as reference genes without confirming their suitability as normalisers. The use of reference genes at random, without regard for stability of expression across treatment groups, can result in erroneous interpretation of data. Here, eleven putative reference genes for use in gene expression studies of the mallard were evaluated, across six different tissues, using a low pathogenic avian influenza A virus infection model. Tissue type influenced the selection of reference genes, whereby different genes were stable in blood, spleen, lung, gastrointestinal tract and colon. β-actin and GAPDH generally displayed low stability and are therefore inappropriate reference genes in many cases. The use of different algorithms (GeNorm and NormFinder) affected stability rankings, but for both algorithms it was possible to find a combination of two stable reference genes with which to normalise qPCR data in mallards. These results highlight the importance of validating the choice of normalising reference genes before conducting gene expression studies in ducks. The fact that nearly all previous studies of the influence of pathogen infection on mallard gene expression have used a single, non-validated reference gene is problematic. The toolkit of putative reference genes provided here offers a solid foundation for future studies of gene expression in mallards and other waterfowl.     

Single cell transcriptomics of neighboring hyphae of <Emphasis Type="Italic">Aspergillus niger</Emphasis>

Single cell profiling was performed to assess differences in RNA accumulation in neighboring hyphae of the fungus Aspergillus niger. A protocol was developed to isolate and amplify RNA from single hyphae or parts thereof. Microarray analysis resulted in a present call for 4 to 7% of the A. niger genes, of which 12% showed heterogeneous RNA levels. These genes belonged to a wide range of gene categories.     

Living with myotonic dystrophy; what can be learned from couples? a qualitative study

Background  

Myotonic dystrophy type 1 (MD1) is one of the most prevalent neuromuscular diseases, yet very little is known about how MD1 affects the lives of couples and how they themselves manage individually and together. To better match health care to their problems, concerns and needs, it is important to understand their perspective of living with this hereditary, systemic disease.     

Approaches for inclusion of forest carbon cycle in life cycle assessment – a review

Forests are a significant pool of terrestrial carbon. A key feature related to forest biomass harvesting and use is the typical time difference between carbon release into and sequestration from the atmosphere. Traditionally, the use of sustainably grown biomass has been considered as carbon neutral in life cycle assessment (LCA) studies. However, various approaches to account for greenhouse gas (GHG) emissions and sinks of forest biomass acquisition and use have also been developed and applied, resulting in different conclusions on climate impacts of forest products. The aim of this study is to summarize, clarify, and assess the suitability of these approaches for LCA. A literature review is carried out, and the results are analyzed through an assessment framework. The different approaches are reviewed through their approach to the definition of reference land‐use situation, consideration of time frame and timing of carbon emissions and sequestration, substitution credits, and indicators applied to measure climate impacts. On the basis of the review, it is concluded that, to account for GHG emissions and the related climate impacts objectively, biomass carbon stored in the products and the timing of sinks and emissions should be taken into account in LCA. The reference situation for forest land use has to be defined appropriately, describing the development in the absence of the studied system. We suggest the use of some climate impact indicator that takes the timing of the emissions and sinks into consideration and enables the use of different time frames. If substitution credits are considered, they need to be transparently presented in the results. Instead of carbon stock values taken from the literature, the use of dynamic forest models is recommended.     

SET8 is degraded via PCNA-coupled CRL4(CDT2) ubiquitylation in S phase and after UV irradiation

The eukaryotic cell cycle is regulated by multiple ubiquitin-mediated events, such as the timely destruction of cyclins and replication licensing factors. The histone H4 methyltransferase SET8 (Pr-Set7) is required for chromosome compaction in mitosis and for maintenance of genome integrity. In this study, we show that SET8 is targeted for degradation during S phase by the CRL4(CDT2) ubiquitin ligase in a proliferating cell nuclear antigen (PCNA)-dependent manner. SET8 degradation requires a conserved degron responsible for its interaction with PCNA and recruitment to chromatin where ubiquitylation occurs. Efficient degradation of SET8 at the onset of S phase is required for the regulation of chromatin compaction status and cell cycle progression. Moreover, the turnover of SET8 is accelerated after ultraviolet irradiation dependent on the CRL4(CDT2) ubiquitin ligase and PCNA. Removal of SET8 supports the modulation of chromatin structure after DNA damage. These results demonstrate a novel regulatory mechanism, linking for the first time the ubiquitin-proteasome system with rapid degradation of a histone methyltransferase to control cell proliferation.     

Beclin1 controls the levels of p53 by regulating the deubiquitination activity of USP10 and USP13

Autophagy is an important intracellular catabolic mechanism that mediates the degradation of cytoplasmic proteins and organelles. We report a potent small molecule inhibitor of autophagy named "spautin-1" for specific and potent autophagy inhibitor-1. Spautin-1 promotes the degradation of Vps34 PI3 kinase complexes by inhibiting two ubiquitin-specific peptidases, USP10 and USP13, that target the Beclin1 subunit of Vps34 complexes. Beclin1 is a tumor suppressor and frequently monoallelically lost in human cancers. Interestingly, Beclin1 also controls the protein stabilities of USP10 and USP13 by regulating their deubiquitinating activities. Since USP10 mediates the deubiquitination of p53, regulating deubiquitination activity of USP10 and USP13 by Beclin1 provides a mechanism for Beclin1 to control the levels of p53. Our study provides a molecular mechanism involving protein deubiquitination that connects two important tumor suppressors, p53 and Beclin1, and a potent small molecule inhibitor of autophagy as a possible lead compound for developing anticancer drugs.