A heterologous in-cell assay for investigating intermicrovillar adhesion complex interactions reveals a novel protrusion length-matching mechanism

Solute transporting epithelial cells build arrays of microvilli on their apical surface to increase membrane scaffolding capacity and enhance function potential. In epithelial tissues such as the kidney and gut, microvilli are length-matched and assembled into tightly packed “brush borders,” which are organized by ∼50-nm thread-like links that form between the distal tips of adjacent protrusions. Composed of protocadherins CDHR2 and CDHR5, adhesion links are stabilized at the tips by a cytoplasmic tripartite module containing the scaffolds USH1C and ANKS4B and the actin-based motor MYO7B. Because several questions about the formation and function of this “intermicrovillar adhesion complex” remain open, we devised a system that allows one to study individual binary interactions between specific complex components and MYO7B. Our approach employs a chimeric myosin consisting of the MYO10 motor domain fused to the MYO7B cargo-binding tail domain. When expressed in HeLa cells, which do not normally produce adhesion complex proteins, this chimera trafficked to the tips of filopodia and was also able to transport individual complex components to these sites. Unexpectedly, the MYO10–MYO7B chimera was able to deliver CDHR2 and CDHR5 to distal tips in the absence of USH1C or ANKS4B. Cells engineered to localize high levels of CDHR2 at filopodial tips acquired interfilopodial adhesion and exhibited a striking dynamic length-matching activity that aligned distal tips over time. These findings deepen our understanding of mechanisms that promote the distal tip accumulation of intermicrovillar adhesion complex components and also offer insight on how epithelial cells minimize microvillar length variability.     

Floristic elements in European vascular plants: an analysis based on Atlas Florae Europaeae

Aim To identify floristic elements in the European flora by an analysis of the distributions of species and species groups mapped in Atlas Florae Europaeae. Location Europe, as defined by Flora Europaea. Methods We analysed the native distributions of 2762 species and 33 species’ aggregates from 79 families, which represent c. 20% of the European flora. The distributional data base, derived from Atlas Florae Europaeae, includes records from 4420 50 × 50‐km UTM grid squares. We classified species into floristic elements by a three‐stage clustering procedure, which consisted of: (1) constructing a dissimilarity hierarchy by complete linkage clustering, using a distance measure based on Jaccard’s coefficient; (2) cutting the hierarchical tree at the 0.95 level to create initial clusters, and forcing small clusters to link with larger ones until the sum of within‐group pairwise distances exceeded a threshold value; and (3) checking the allocation of all species to the redefined clusters and reassigning species if appropriate, using the cosine of the angle between the species and cluster centres to measure the similarity of species to clusters. Results The clustering procedure classified 2793 taxa into 18 floristic elements, which included between 66 and 289 taxa; two species had unique, non‐overlapping distributions and could not be classified. Main conclusions The analysis highlights the floristic diversity of the mountains of central and southern Europe, and of the Mediterranean region. The floristic elements of northern latitudes and the temperate lowlands tend to be composed of wide‐ranging species and include only a low proportion of European endemics. The montane elements, including those centred on montane areas in the Mediterranean region, are composed predominantly of perennial species and include high or very high proportions of European endemics. Classifications that recognize one ‘Alpine’ and one ‘Mediterranean’ biogeographical zone in Europe fail to reflect this floristic diversity.     

BRCT domains: Easy as one,two, three

BRCA1 C-terminal (BRCT) domains are integral signaling modules in the DNA damage response (DDR). Aside from their established roles as phospho-peptide binding modules, BRCT domains have been implicated in phosphorylation-independent protein interactions, DNA binding and poly(ADP-ribose) (PAR) binding. These numerous functions can be attributed to the diversity in BRCT domain structure and architecture, where domains can exist as isolated single domains or assemble into higher order homo- or hetero-domain complexes. In this review, we incorporate recent structural and biochemical studies to demonstrate how structural features allow single and tandem BRCT domains to attain a high degree of functional diversity.Key words: BRCT domain, DNA repair, phosphorylation, phospho-peptide interaction, protein interaction, DNA binding, DNA damage response     

Reduced scent marking and ultrasonic vocalizations in the BTBR T+tf/J mouse model of autism

Qualitative impairments in communication, such as delayed language and poor interactive communication skills, are fundamental to the diagnosis of autism. Investigations into social communication in adult BTBR T+tf/J (BTBR) mice are needed to determine whether this inbred strain incorporates phenotypes relevant to the second diagnostic symptom of autism, communication deficits, along with its strong behavioral phenotypes relevant to the first and third diagnostic symptoms, impairments in social interactions and high levels of repetitive behavior. The aim of the present study was to simultaneously measure female urine‐elicited scent marking and ultrasonic vocalizations in adult male BTBR mice, in comparison with a standard control strain with high sociability, C57BL/6J (B6), for the assessment of a potential communication deficit in BTBR. Adult male BTBR mice displayed lower scent marking and minimal ultrasonic vocalization responses to female urine obtained from both B6 and BTBR females. Lower scent marking and ultrasonic vocalizations in a social setting by BTBR, as compared with B6, are consistent with the well‐replicated social deficits in this inbred mouse strain. Our findings support the interpretation that BTBR incorporate communication deficits, and suggest that scent marking and ultrasonic vocalizations offer promising measures of interest in social cues that may be widely applicable to investigations of mouse models of autism.     

Feasibility of outpatient management after intra-articular yttrium-90: comparison of two regimens.

In a study comparing two regimens of treatment after intra-articular irradiation of the knee with yttrium-90 one group of patients was allocated to bed rest for 48 hours in hospital and the other to mobilisation at home. Initially a Robert-Jones orthopaedic bandage was applied to the knee in all patients, serving as a semi-rigid splint, but as loss of isotope from the knee was appreciable in the mobilised patients, subsequent patients were sent home with the knee in a plaster-of-Paris cylinder. No difference in extra-articular spread or chromosomal damage was found between the patients sent home with their knee in a rigid splint and those treated by bed rest. Clinical outcome at three months was satisfactory in all three groups. These results show that rigid splinting is essential in reducing extra-articular spread of the isotope but that bed rest is not necessary. Increases in intra-articular pressure associated with quadriceps muscle activity combined with flexion of the knee may be the most important factor affecting extra-articular spread of isotope.     

Pyrrole[2,3-d]azepino compounds as agonists of the farnesoid X receptor (FXR)

Pyrrole[2,3-d]azepines have been identified as potent agonists of the farnesoid X receptor (FXR). Based on the planar X-ray crystal structure of WAY-362450 1 in the ligand binding domain and molecular modeling studies, non-planar reduced compounds were designed which led to agonists that exhibit high aqueous solubility and retain moderate in vitro potency.     

Orosensory detection of fatty acids by obesity-prone and obesity-resistant rats: strain and sex differences

A series of brief-access (15s) behavioral assays following the formation of a conditioned taste aversion (CTA) to linoleic acid were performed in order to follow up on observations showing differences in the chemosensory responses to dietary fat in obesity-prone (Osborne-Mendel [O-M]) and obesity-resistant (S5B/Pl) rat strains. Strong aversions to linoleic acid (conditioned stimulus 100 microM) were generated in both O-M and S5B/Pl rats to concentrations as low as 2.5 microM. Observed strain differences were in contrast to expectations based upon electrophysiological studies previously showing greater fatty acid-induced inhibition of delayed rectifying K+ channels in S5B/Pl rats. In the CTA assays, the O-M rats showed aversions at lower fatty acid concentrations with more resistance to extinction in brief-access orosensory tests, suggesting that the obesity-prone strain may be more sensitive in the detection and subsequent avoidance of linoleic acid than the obesity-resistant strain. The independent variable of sex produced even greater differences in the avoidance of linoleic acid following conditioning than the effects of strain. Female rats of both strains were significantly more sensitive to fatty acids, showed greater cross-generalization from linoleic to oleic acid, and showed greater avoidance of linoleic acid than male counterparts. These findings suggest genetic influences on yet to be identified mechanisms potentially within the gustatory system that affect the sensitivity to detect the fatty acid chemicals found in dietary fat during brief-access orosensory testing.