A Conditionally Fluorescent Peptide Reporter of Secondary Structure Modulation

Proteins containing intrinsic disorder often form secondary structure upon interaction with a binding partner. Modulating such structures presents an approach for manipulating the resultant functional outcomes. Translational repressor protein 4E-BP1 is an example of an intrinsically disordered protein that forms an α-helix upon binding to its protein ligand, eIF4E. Current biophysical methods for analyzing binding-induced structural changes are low-throughput, require large amounts of sample, or are extremely sensitive to signal interference by the ligand itself. Herein, we describe the discovery and development of a conditionally fluorescent 4E-BP1 peptide that reports structural changes of its helix in high-throughput format. This reporter peptide is based on conditional quenching of fluorescein by thioamides. In this case, fluorescence signal increases as the peptide becomes more ordered. Conversely, destabilization of the α-helix results in decreased fluorescence signal. The low concentration and low volume of peptide required make this approach amenable for high-throughput screening to discover ligands that alter peptide secondary structure.     

The distribution,density, and biomass of the zebra mussel (Dreissena polymorpha) on natural substrates in Lake Winnipeg 2017–2019

The distribution, density, biomass and size-structure of the zebra mussel (Dreissena polymorpha) population in Lake Winnipeg were examined between 2017 and 2019. Zebra mussels have colonized most of the available hard substrate in the south basin and Narrows region, but colonization of the north basin remains low at present, even on suitable substrate. Numerical densities and shell free biomass peaked at 5530 ± 953 m^?2 and 64.7 ± 57.9 g shell free dry mass m^?2 respectively. The distribution appeared to be strongly limited by substrate type and availability, with further limitations on the distribution imposed by physical disturbance in shallow waters and unsuitable substrate in deeper areas of the lake. Zebra mussels <1 year old dominated the populations, and individuals >18 mm were exceedingly rare. Poor recruitment was observed at sites along the eastern side of the south basin compared to elsewhere in the lake. The proximate causes of these differences in colonization success and recruitment are not clear, but may be in part due to heterogeneous patterns of key physico-chemical environmental conditions such as calcium concentrations required for successful development of juvenile mussels and colder water temperatures in the north basin. This study provides a baseline of information on which to track further expansion of zebra mussels in Lake Winnipeg and assist efforts to develop an understanding of how zebra mussels may affect the ecology of Lake Winnipeg.     

A microfluidic toolbox for cell fusion

Cellular fusion is a key process in many fields ranging from historical gene mapping studies and monoclonal antibody production, through to cell reprogramming. Traditional methodologies for cell fusion rely on the random pairing of different cell types and generally result in low and variable fusion efficiencies. These approaches become particularly limiting where substantial numbers of bespoke one‐to‐one fusions are required, for example, for in‐depth studies of nuclear reprogramming mechanisms. In recent years, microfluidic technologies have proven valuable in creating platforms where the manipulation of single cells is highly efficient, rapid and controllable. These technologies also allow the integration of different experimental steps and characterisation processes into a single platform. Although the application of microfluidic methodologies to cell fusion studies is promising, current technologies that rely on static trapping are limited both in terms of the overall number of fused cells produced and their experimental accessibility. Here we review some of the most exciting breakthroughs in core microfluidic technologies that will allow the creation of integrated platforms for controlled cell fusion at high throughput. © 2015 Society of Chemical Industry     

Characterization of Microencapsulated Rosemary Essential Oil and Its Antimicrobial Effect on Fresh Dough

This study aimed to characterize rosemary essential oil particles obtained by spray-drying and to evaluate their antimicrobial activity. Measurements of mycelial growth of Penicillium and Aspergillus fungi, isolated from fresh dough, were made applying oil concentrations of 1.0, 5.0, 10.0, 20.0, and 50.0 μL/mL. Fungi and yeast counts in the fresh dough submitted to the control (no oil), pure oil, and microencapsulated oil treatments were also conducted. The microcapsules showed 50 % oil retention after drying, and the major oil components identified were 1,8-cineole (29.0 %), camphor (26.6 %), and α-pinene (10.6 %). The analysis of the particles revealed surfaces without fissures, with a mean particle size of 12.2 μm and presenting an amorphous structure. The growth inhibiting effect of Penicillium sp. fungus, compared to the control, was verified at concentrations of 1.0, 5.0, and 10.0 μL/mL rosemary essential oil, which did not differ among them. For Aspergillus sp., the application of 10.0 μL/mL oil provided greater inhibition compared to 1.0 and 5.0 μL/mL. Complete inhibition occurred with the application of 50.0 μL/mL for both fungal genera tested. At 8 days of dough storage at 25 °C, a decrease of at least 0.7 and 1.5 log cycles of fungal growth was observed in the dough with pure oil and that with microencapsulated oil, respectively, relative to the control. The microencapsulation process retained the antimicrobial property of rosemary essential oil and provided further extension of this activity over time when applied to fresh dough.     

Variation in Preen Oil Composition Pertaining to Season,Sex, and Genotype in the Polymorphic White-Throated Sparrow

Evidence for the the ability of birds to detect olfactory signals is now well documented, yet it remains unclear whether birds secrete chemicals that can be used as social cues. A potential source of chemical cues in birds is the secretion from the uropygial gland, or preen gland, which is thought to waterproof, maintain, and protect feathers from ectoparasites. However, it is possible that preen oil also may be used for individual recognition, mate choice, and signalling social/sexual status. If preen oil secretions can be used as socio-olfactory signals, we should be able to identify the volatile components that could make the secretions more detectable, determine the seasonality of these secretions, and determine whether olfactory signals differ among relevant social groups. We examined the seasonal differences in volatile compounds of the preen oil of captive white-throated sparrows, Zonotrichia albicollis. This species is polymorphic and has genetically determined morphs that occur in both sexes. Mating is almost exclusively disassortative with respect to morph, suggesting strong mate choice. By sampling the preen oil from captive birds in breeding and non-breeding conditions, we identified candidate chemical signals that varied according to season, sex, morph, and species. Linear alcohols with a 10–18 carbon chains, as well as methyl ketones and carboxylic acids, were the most abundant volatile compounds. Both the variety and abundances of some of these compounds were different between the sexes and morphs, with one morph secreting more volatile compounds in the non-breeding season than the other. In addition, 12 compounds were seasonally elevated in amount, and were secreted in high amounts in males. Finally, we found that preen oil signatures tended to be species-specific, with white-throated sparrows differing from the closely related Junco in the abundances and/or prevalence of at least three compounds. Our data suggest roles for preen oil secretions and avian olfaction in both non-social as well as social interactions.     

Peaceful,Pleasant and Private: The British Domestic Garden as an Ordinary Landscape

This paper uses narrative accounts of private gardens in Britain from the Mass-Observation Archive (MO) to explore ideas of landscape, privacy and attachment that emerge from daily practices and routines in these ordinary domestic spaces. We argue for the domestic garden as a vernacular or ordinary landscape that displays tensions between the private and the public nature of home within ambivalent emotional responses. Extended personal narratives offer privileged access to a site of intense engagement and carefully guarded privacy, yet with varying levels of attachment. The garden is a space well described in Britain in its public form but less well known as a private, everyday landscape. In this way a cultural landscape study becomes a contemporary critical geography of an ordinary space.     

Structure–Property Relationships in Aligned Electrospun Barium Titanate Nanofibers

Barium titanate nanofibers were uniaxially aligned by electrospinning onto a rotating copper wire drum and alignment was maintained during calcination of the fibers. Two methods for maintaining alignment during calcination were tested, by either using carbon tape or a peeling off method to remove the aligned fibers from the mandrel followed by calcination. The carbon tape removal method led to the formation of shorter aligned nanowires while the peeling off method resulted in longer nanofibers. Additionally, the effects of calcination temperature and time on crystal structure were also examined. The degree of tetragonality in the barium titanate nanofibers increased at higher calcination temperatures and times. Piezoelectricity was confirmed in the nanofibers calcined using piezoeresponse force microscopy, yielding a d₃₃ value of 15.5 pm/V. Using the methods presented here, large quantities of aligned piezoelectric barium titanate and other ceramic fibers or wires can be produced to fulfill their demand in novel microelectronics.