Identification of 51 novel exons of the Usher syndrome type 2A (USH2A) gene that encode multiple conserved functional domains and that are mutated in patients with Usher syndrome type II

The USH2A gene is mutated in patients with Usher syndrome type IIa, which is the most common subtype of Usher syndrome and is characterized by hearing loss and retinitis pigmentosa. Since mutation analysis by DNA sequencing of exons 1-21 revealed only ~63% of the expected USH2A mutations, we searched for so-far-uncharacterized exons of the gene. We identified 51 novel exons at the 3' end of the gene, and we obtained indications for alternative splicing. The putative protein encoded by the longest open reading frame harbors, in addition to the known functional domains, two laminin G and 28 fibronectin type III repeats, as well as a transmembrane region followed by an intracellular domain with a PDZ-binding domain at its C-terminal end. Semiquantitative expression profile analysis suggested a low level of expression for both the long and the short isoform(s) and partial overlap in spatial and temporal expression patterns. Mutation analysis in 12 unrelated patients with Usher syndrome, each with one mutation in exons 1-21, revealed three different truncating mutations in four patients and two missense mutations in one patient. The presence of pathogenic mutations in the novel exons indicates that at least one of the putative long isoforms of the USH2A protein plays a role in both hearing and vision.     

Environmental enrichment: room for reduction?

Environmental enrichment strategies are usually regarded as refinement. However, when the welfare of animals is enhanced through successful enrichment programmes, a reduction in the number of animals needed can be expected, because fewer animals might be lost during the course of experiments. Several examples of studies where enrichment can lead to reduction will be presented. They include the beneficial effects of nesting material for laboratory mice, the effects of husbandry procedures on controlling aggressive behaviour in male laboratory mice, and the effects of enrichment on variation in the results of experiments.     

Moderate Intense Physical Activity Depends on Selected Metabolic Equivalent of Task (MET) Cut-Off and Type of Data Analysis

Background

Accelerometry data are frequently analyzed without considering whether moderate-to-vigorous physical activities (MVPA) were performed in bouts of >10 minutes as defined in most physical activity guidelines. We aimed i) to quantify MVPA by using different commonly-applied physical activity guidelines, ii) to investigate the effect of bouts versus non-bouts analysis, and iii) to propose and validate a MVPA non-bouts cut-point to classify (in-) active subjects.

Methods

Healthy subjects (n=110;62±6yrs) and patients with Chronic Obstructive Pulmonary Disease (COPD) (n=113;62±5yrs) wore an activity monitor for 7 days. Three Metabolic Equivalent of Task (MET) cut-offs and one individual target (50% VO₂ reserve) were used to define MVPA. First, all minutes of MVPA were summed up (NON-BOUTS). Secondly, only minutes performed in bouts of >10 minutes continuous activity were counted (BOUTS). Receiver operating characteristic (ROC) curve analyses were used to propose and (cross-) validate new MVPA non-bout cut-points based on the criterion of 30 minutes MVPA per day (BOUTS). Likelihood ratios (sensitivity/[1-specificity]) were used to express the association between the proposed MVPA non-bout target and the MVPA bout target of 30 min*day^-1.

Results

MVPA was variable across physical activity guidelines with lowest values for age-specific cut-offs. Selecting a METs cut-point corresponding to 50% VO₂ reserve revealed no differences in MVPA between groups. MVPA’s analyzed in BOUTS in healthy subjects were 2 to 4 fold lower than NON-BOUTS analyses and this was even 3 to 12 fold lower in COPD. The MVPA non-bouts cut-point of 80 min*day^-1 using a 3 METs MVPA threshold delivered positive likelihood ratios of 5.1[1.5-19.6] (healthy subjects) and 2.3[1.6-3.3] (COPD).

Conclusion

MVPA varies upon the selected physical activity guideline/targets and bouts versus non-bouts analysis. Accelerometry measured MVPA non-bouts target of 80 min*day^-1, using a 3 METs MVPA threshold, is associated to the commonly-used MVPA bout target of 30 min*day^-1.     

Climate warming feedback from mountain birch forest expansion: reduced albedo dominates carbon uptake

Expanding high‐elevation and high‐latitude forest has contrasting climate feedbacks through carbon sequestration (cooling) and reduced surface reflectance (warming), which are yet poorly quantified. Here, we present an empirically based projection of mountain birch forest expansion in south‐central Norway under climate change and absence of land use. Climate effects of carbon sequestration and albedo change are compared using four emission metrics. Forest expansion was modeled for a projected 2.6 °C increase in summer temperature in 2100, with associated reduced snow cover. We find that the current (year 2000) forest line of the region is circa 100 m lower than its climatic potential due to land‐use history. In the future scenarios, forest cover increased from 12% to 27% between 2000 and 2100, resulting in a 59% increase in biomass carbon storage and an albedo change from 0.46 to 0.30. Forest expansion in 2100 was behind its climatic potential, forest migration rates being the primary limiting factor. In 2100, the warming caused by lower albedo from expanding forest was 10 to 17 times stronger than the cooling effect from carbon sequestration for all emission metrics considered. Reduced snow cover further exacerbated the net warming feedback. The warming effect is considerably stronger than previously reported for boreal forest cover, because of the typically low biomass density in mountain forests and the large changes in albedo of snow‐covered tundra areas. The positive climate feedback of high‐latitude and high‐elevation expanding forests with seasonal snow cover exceeds those of afforestation at lower elevation, and calls for further attention of both modelers and empiricists. The inclusion and upscaling of these climate feedbacks from mountain forests into global models is warranted to assess the potential global impacts.     

Movement and vertical stratification of fruit-feeding butterflies in a managed West African rainforest

We analysed movement parameters and vertical stratification in fruit-feeding butterflies between a control, a thinned and a plantation site within a West African rainforest. Overall, distances moved between traps were largest in the plantation. Movement parameters were generally largest in species feeding on early successional hostplants of gap and margin habitats. In these species, distances between recaptures were significantly shorter in the thinned compared to the control forest. Conversely, forest floor species feeding on climbers and understorey shrubs showed significantly larger movement in the thinned forest. Higher strata species also flew larger distances in the thinned understorey. Including higher strata samples, they were significantly less abundant in the thinned plot, although understorey sampling alone indicated the contrary. Comparing vertical distribution patterns between thinned and control sites indicated a disruption of vertical stratification after thinning. Canopy species seem to fly in the upper strata of the more closed-canopy control forest, whereas they descend more frequently in the forest opened by the thinning management. Understorey sampling might therefore lead to biased conclusions due to differences in vertical distribution between forest plots. This study showed that thinning can affect the restricted-range forest floor butterflies as well as the more widespread canopy butterfly fauna.     

Human immature dendritic cells efficiently bind and take up secretory IgA without the induction of maturation.

Immature dendritic cells (DC) reside in peripheral tissues, where they pick up and process incoming pathogens via scavenger receptors or FcR such as FcgammaR and FcepsilonR. At mucosal surfaces, IgA is the main Ig to protect the body from incoming pathogens. In addition, DC are present in high numbers at these sites. We detected expression of FcalphaR (CD89) on the CD14+ population of CD34+ progenitor-derived DC and on monocyte-derived DC (MoDC). However, CD89 expression was strongly decreased upon differentiation from monocyte to DC. We found only minimal binding of serum IgA to MoDC but strong binding of secretory IgA (SIgA). The SIgA binding to MoDC could not be blocked by anti-CD89 blocking Abs. DC efficiently internalized SIgA, but not serum IgA, and uptake of SIgA could be blocked by specific sugars or partially by Ab reactive with mannose receptor. Importantly, binding and uptake of SIgA was not accompanied by signs of DC maturation, such as increased expression of CD86 and CD83 or induction of cytokine secretion. These data indicate that SIgA can interact with DC not via CD89, but via carbohydrate-recognizing receptors like mannose receptor and suggest that uptake of SIgA-containing immune complexes by immature DC may be a mechanism to modulate mucosal immune responses.     

The N-terminal -barrel domain of the Escherichia coli K88 periplasmic chaperone FaeE determines fimbrial subunit recognition and dimerization

The K88 periplasmic chaperone FaeE is a homodimer, whereas the K99 chaperone FanE is a monomer. The structural requirements for dimerization of the K88 fimbrial periplasmic chaperone and for fimbrial subunit-binding specificity were investigated by analysis of mutant chaperones. FaeE contains a C-terminal extension of 19 amino acid residues when compared to FanE and most other fimbrial chaperones. A C-terminal truncate of the K88 chaperone FaeE was constructed that lacked 19 C-terminal amino acid residues. Expression and complementation experiments revealed that this C-terminal shortened chaperone was still functional in binding the K88 major subunit FaeG and K88 biosynthesis. Two hybrid chaperones were constructed. Each hybrid protein contained one -barrel domain of FaeE and the other -barrel domain of FanE (Fae/FanE or Fan/FaeE, respectively). Expression and complementation experiments revealed that the Fae/FanE but not the Fan/FaeE hybrid chaperone was functional in the formation of K88 fimbriae. The Fan/FaeE hybrid chaperone was active in the biosynthesis of K99 fimbriae. The truncated FaeE mutant chaperone and the hybrid Fae/FanE chaperone were able to form stable periplasmic protein complexes with the K88 major fimbrial subunit FaeG. Cross-linking experiments suggested that the C-terminal shortened chaperone and the Fae/FanE hybrid chaperone were homodimers, as is the wild-type K88 chaperone. Altogether, the data suggested that the N-terminal -barrel domain of a fimbrial chaperone determines subunit specificity. In the case of the K88 periplasmic chaperone, this N-terminal domain also determines dimerization of the protein.     

Therapist-supported Internet-based cognitive behaviour therapy yields similar effects as face-to-face therapy for psychiatric and somatic disorders: an updated systematic review and meta-analysis

Providing therapist-guided cognitive behaviour therapy via the Internet (ICBT) has advantages, but a central research question is to what extent similar clinical effects can be obtained as with gold-standard face-to-face cognitive behaviour therapy (CBT). In a previous meta-analysis published in this journal, which was updated in 2018, we found evidence that the pooled effects for the two formats were equivalent in the treatment of psychiatric and somatic disorders, but the number of published randomized trials was relatively low (n=20). As this is a field that moves rapidly, the aim of the current study was to conduct an update of our systematic review and meta-analysis of the clinical effects of ICBT vs. face-to-face CBT for psychiatric and somatic disorders in adults. We searched the PubMed database for relevant studies published from 2016 to 2022. The main inclusion criteria were that studies had to compare ICBT to face-to-face CBT using a randomized controlled design and targeting adult populations. Quality assessment was made using the Cochrane risk of bias criteria (Version 1), and the main outcome estimate was the pooled standardized effect size (Hedges’ g) using a random effects model. We screened 5,601 records and included 11 new randomized trials, adding them to the 20 previously identified ones (total n=31). Sixteen different clinical conditions were targeted in the included studies. Half of the trials were in the fields of depression/depressive symptoms or some form of anxiety disorder. The pooled effect size across all disorders was g=0.02 (95% CI: –0.09 to 0.14) and the quality of the included studies was acceptable. This meta-analysis further supports the notion that therapist-supported ICBT yields similar effects as face-to-face CBT.