Genome sequence and genetic linkage analysis of Shiitake mushroom _Lentinula edodes_

_Lentinula edodes_ (Shiitake/Xianggu) is an important cultivated mushroom. Understanding the genomics and functional genomics of _L. edodes_ allows us to improve its cultivation and quality. Genome sequence is a key to develop molecular genetic markers for breeding and genetic manipulation. We sequenced the genome of _L. edodes_ monokaryon L54A using Roche 454 and ABI SOLiD genome sequencing. Sequencing reads of about 1400Mb were de novo assembled into a 40.2 Mb genome sequence. We compiled the genome sequence into a searchable database with which we have been annotating the genes and analyzing the metabolic pathways. In addition, we have been using many molecular techniques to analyze genes differentially expressed during development. Gene ortholog groups of _L. edodes_ genome sequence compared across genomes of several fungi including mushrooms identified gene families unique to mushroom-forming fungi. We used a mapping population of haploid basidiospores of dikaryon L54 for genetic linkage analysis. High-quality variations such as single nucleotide polymorphisms, insertions, and deletions of the mapping population formed a high-density genetic linkage map. We compared the linkage map to the _L. edodes_ L54A genome sequence and located selected quantitative trait loci. The Shiitake community will benefit from these resources for genetic studies and breeding.
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Evaluation of the ClearVoice Strategy in Adults Using HiResolution Fidelity 120 Sound Processing

ObjectivesThis study aimed to evaluate the benefits of ClearVoice strategy on speech perception in noise and in everyday listening situations in Cantonese-speaking cochlear implant users.MethodsTwelve experienced adult users of the Harmony implant and HiRes 120 sound processing participated in the study. The study employed a prospective within-subjects design wherein speech recognition in adults using HiRes 120 without ClearVoice turned on (control option) was compared to their performance with HiRes 120 with ClearVoice turned on. Each subject was evaluated with two different ClearVoice gain settings: -12 dB (ClearVoice medium) and -18 dB (ClearVoice high) after one-week of use. The Cantonese hearing in noise test and a questionnaire were used as the outcome measures.ResultsSubjects performed significantly better with ClearVoice medium than with control option in noise. No significant difference in performance was noted among the 3 settings in quiet. Most subjects reported high level of satisfaction with ClearVoice in daily listening situations and preferred to keep ClearVoice on.ConclusionClearVoice can help cochlear implant recipients to hear better in noise

Mindfulness-based cognitive therapy v. group psychoeducation for people with generalised anxiety disorder: randomised controlled trial

Background: Research suggests that an 8-week mindfulness-based cognitive therapy (MBCT) course may be effective for generalised anxiety disorder (GAD). Aims: To compare changes in anxiety levels among participants with GAD randomly assigned to MBCT, cognitive–behavioural therapy-based psychoeducation and usual care. Method: In total, 182 participants with GAD were recruited (trial registration number: CUHK_CCT00267) and assigned to the three groups and followed for 5 months after baseline assessment with the two intervention groups followed for an additional 6 months. Primary outcomes were anxiety and worry levels. Results: Linear mixed models demonstrated significant group × time interaction (F(4,148) = 5.10, P = 0.001) effects for decreased anxiety for both the intervention groups relative to usual care. Significant group × time interaction effects were observed for worry and depressive symptoms and mental health-related quality of life for the psychoeducation group only. Conclusions: These results suggest that both of the interventions appear to be superior to usual care for the reduction of anxiety symptoms

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Protective effect of herba cistanches against oxidant injury

Earlier findings have demonstrated that pretreatment of Herba Cistanches, a Yang-invigorating Chinese tonic herb, stimulated the ATP-generation capacity (ATP-GC) in mitochondria isolated from rat hearts ex vivo. The enhancement of mitochondrial ATP-GC by Herba Cistanches was associated with the induction of mitochondrial uncoupling and the protection against ischemic/reperfusion (I/R) injury in rat hearts. This thesis study aimed at (1) identifying the active fraction(s) in Herba Cistanches and (2) further investigating its biological activities and their underlying mechanism. Using ATP-GC stimulation as an activity monitor, a semi-purified fraction of Herba Cistanches ethanol extract (HCF1), which contained relatively non-polar ingredients, was isolated and subjected to further investigations. Pretreatment of HCF1 was found to increase ATP-GC and induce mitochondrial uncoupling as well as cellular antioxidant response in H9c2 cardiomyocytes, with the latter being assessed by the enhancement of glutathione reductase (GR)-mediated glutathione redox cycling. It was hypothesized that the increase in mitochondrial reactive oxygen species (ROS) production associated with the stimulation of ATP-GC can trigger cellular responses including mitochondrial uncoupling and GR-mediated glutathione redox cycling, with resultant protection against oxidant injury. To test this hypothesis, the effect of HCF1 on mitochondrial ROS production was examined. HCF1 pretreatment increased mitochondrial ROS generation in H9c2 cardiomyocytes. The removal of mitochondrial ROS by antioxidants (dimethylthiourea and Trolox) abrogated the HCF1-induced cellular responses and the associated cytoprotective effect. Studies using specific inhibitors of reduced glutathione synthesis and regeneration suggested the important role of GR-mediated glutathione redox cycling in the cytoprotection afforded by HCF1. Results obtained from animal studies showed that HCF1 pretreatment protected against myocardial I/R injury in rats. In conclusion, HCF1, an active fraction of Herba Cistanches that contained relatively non-polar ingredients, could enhance mitochondrial ATP-GC in H9c2 cardiomyocytes and mitochondria isolated from rat heart. The associated increase in mitochondrial ROS production induced mitochondrial uncoupling and GR-mediated glutathione redox cycling in H9c2 cardiomyocytes. The enhancement of glutathione redox cycling by HCF1 protected against oxidant injury in cardiomyocytes. The long-term intake of HCF1 can offer a prospect for the prevention of cardiovascular diseases

Are mitochondria the main contributor of reactive oxygen species in cells

Physiologists often assume that mitochondria are the main producers of reactive oxygen species (ROS) in cells. Consequently, in biomedicine, mitochondria are considered as important targets for therapeutic treatments, and in evolutionary biology, they are considered as mediators of life-history tradeoffs. Surprisingly, data supporting such an assumption are lacking, at least partially due to the technical difficulties in accurately measuring the level of ROS produced by different subcellular compartments in intact cells. In this Commentary, we first review three potential reasons underlying the misassumption of mitochondrial dominance in the production of cellular ROS. We then introduce some other major sites/enzymes responsible for cellular ROS production. With the use of a recently developed cell-based assay, we further discuss the contribution of mitochondria to the total rate of ROS release in cell lines and primary cells of different species. In these cells, the contribution of mitochondria varies between cell types but mitochondria are never the main source of cellular ROS. This indicates that although mitochondria are one of the significant sources of cellular ROS, they are not necessarily the main contributor under normal conditions. Intriguingly, similar findings were also observed in cells under a variety of stressors, life-history strategies and pathological stages, in which the rates of cellular ROS production were significantly enhanced. Finally, we make recommendations for designing future studies. We hope this paper will encourage investigators to carefully consider non-mitochondrial sources of cellular ROS in their study systems or models

Peer coaching in action research as a lived practice for teacher professional development

abstractpublished_or_final_versionEducationMasterMaster of Educatio

Mitochondrial and cytosolic sources of hydrogen peroxide in resting C2C12 myoblasts

International audienc