Light scattering corrections to linear dichroism spectroscopy for liposomes in shear flow using calcein fluorescence and modified Rayleigh-Gans-Debye-Mie scattering

The interpretation of data from absorbance spectroscopy experiments of liposomes in flow systems is often complicated by the fact that there is currently no easy way to account for scattering artefacts. This has proved particularly problematic for linear dichroism (LD) spectroscopy, which may be used to determine binding modes of small molecules, peptides and proteins to liposomes if we can extract the absorbance signal from the combined absorbance/scattering experiment. Equations for a modified Rayleigh-Gans-Debye (RGD) approximation to the turbidity (scattering) LD spectrum are available in the literature though have not been implemented. This review summarises the literature and shows how it can be implemented. The implementation proceeds by first determining volume loss that occurs when a spherical liposome is subjected to flow. Calcein fluorescence can be used for this purpose since at high concentrations (>?60 mM) it has low intensity fluorescence with maxima at 525 and 563 nm whereas at low concentrations (<1 mM) the fluorescence intensity is enhanced and the band shifts to 536 nm. The scattering calculation process yields the average axis ratios of the distorted liposome ellipsoids and extent of orientation of the liposomes in flow. The scattering calculations require methods to estimate liposome integrity, volume loss, and orientation when subjected to shear stresses under flow.     

Multi-centre randomised controlled trial of a smart phone-based event recorder alongside standard care versus standard care for patients presenting to the Emergency Department with palpitations and pre-syncope - the IPED (Investigation of Palpitations in the ED) study: study protocol for a randomised controlled trial

Background

Palpitations and pre-syncope are together responsible for 300,000 annual Emergency Department (ED) attendances in the United Kingdom (UK). Diagnosis of the underlying rhythm is difficult as many patients are fully recovered on ED arrival; and examination and presenting electrocardiogram (ECG) are commonly normal. The only way to establish the underlying heart rhythm is to capture an ECG during symptoms. Recent technology advances have led to several novel ECG monitoring devices appearing on the market. This trial aims to compare the symptomatic rhythm detection rate at 90?days of one such smart phone-based event recorder (AliveCor Heart Monitor and AliveECG) with standard care for participants presenting to the ED with palpitations and pre-syncope and no obvious cause in the ED.

Methods/Design

This is a multi-centre hospital ED / Acute Medical Unit (AMU) open label, randomised controlled trial. Participants will be recruited in 10 tertiary and district general hospitals in the UK. Participants aged ≥?16?years presenting with an episode of palpitations or pre-syncope with no obvious cause and whose underlying ECG rhythm during these episodes remains undiagnosed after clinical assessment will be included. Participants will be randomised to either: (1) the intervention arm, standard care plus the use of a smart phone-based event recorder; or (2) the control arm, standard care. Primary endpoint will be symptomatic rhythm detection rate at 90?days. A number of secondary clinical, process and cost-effectiveness endpoints will be collected and analysed. Analysis will be on an intention-to-treat basis.

Discussion

The Investigation of Palpitations in the ED (IPED) study aims to recruit 242 participants across 10 hospital sites. It will be the first study to investigate the ability of a smart phone-based event recorder to detect symptomatic cardiac rhythms compared to standard care for ED patients with palpitations and pre-syncope with no obvious cause in the ED. This smart phone event recorder will allow ED patients who have presented with palpitations or pre-syncope to record their ECG tracing if they have a further episode and may increase the rate of underlying rhythm diagnosis.

Trial registration

ClinicalTrials.gov, NCT02783898. Registered on 26 May 2016.

     

Cellulose synthase ‘class specific regions’ are intrinsically disordered and functionally undifferentiated

Cellulose synthases(CESAs) are glycosyltransferases that catalyze formation of cellulose microfibrils in plant cell walls.Seed plant CESA isoforms cluster in six phylogenetic clades, whose non-interchangeable members play distinct roles within cellulose synthesis complexes(CSCs).A ‘class specific region’(CSR), with higher sequence similarity within versus between functional CESA classes, has been suggested to contribute to specific activities or interactions of different isoforms.We investigated CESA isoform specificity in the moss,Physcomitrella patens(Hedw.) B.S.G.to gain evolutionary insights into CESA structure/function relationships.Like seed plants, P.patens has oligomeric rosette-type CSCs, but the Pp CESAs diverged independently and form a separate CESA clade.We showed that P.patens has two functionally distinct CESAs classes, based on the ability to complement the gametophore-negative phenotype of a ppcesa5 knockout line.Thus, non-interchangeable CESA classes evolved separately in mosses and seed plants.However, testing of chimeric moss CESA genes for complementation demonstrated that functional classspecificity is not determined by the CSR.Sequence analysis and computational modeling showed that the CSR is intrinsically disordered and contains predicted molecular recognition features, consistent with a possible role in CESA oligomerization and explaining the evolution of class-specific sequences without selection for class-specific function.     

Male mate choice via cuticular hydrocarbon pheromones drives reproductive isolation between Drosophila species

Mate discrimination is a key mechanism restricting gene flow between species. While studied extensively with respect to female mate choice, mechanisms of male mate choice between species are far less studied. Thus, we have little knowledge of the relative frequency, importance, or overall contribution of male mate discrimination to reproductive isolation. In the present study, we estimated the relative contributions of male and female choice to reproductive isolation between Drosophila simulans and D. sechellia, and show that male mate discrimination accounts for the majority of the current isolation between these species. We further demonstrate that males discriminate based on female cuticular hydrocarbon pheromones, and collect evidence supporting the hypothesis that male mate discrimination may alleviate the costs associated with heterospecific courtship and mating. Our findings highlight the potentially significant contribution of male mate choice to the formation of reproductive isolating barriers, and thus the speciation process.     

Assessing global range expansion in a cryptic species complex: insights from the red seaweed genus Asparagopsis (Florideophyceae)

The mitochondrial genetic diversity, distribution and invasive potential of multiple cryptic operational taxonomic units (OTUs) of the red invasive seaweed Asparagopsis were assessed by studying introduced Mediterranean and Hawaiian populations. Invasive behavior of each Asparagopsis OTU was inferred from phylogeographic reconstructions, past historical demographic dynamics, recent range expansion assessments and future distributional predictions obtained from demographic models. Genealogical networks resolved Asparagopsis gametophytes and tetrasporophytes into four A. taxiformis and one A. armata cryptic OTUs. Falkenbergia isolates of A. taxiformis L3 were recovered for the first time in the western Mediterranean Sea and represent a new introduction for this area. Neutrality statistics supported past range expansion for A. taxiformis L1 and L2 in Hawaii. On the other hand, extreme geographic expansion and an increase in effective population size were found only for A. taxiformis L2 in the western Mediterranean Sea. Distribution models predicted shifts of the climatically suitable areas and population expansion for A. armata L1 and A. taxiformis L1 and L2. Our integrated study confirms a high invasive risk for A. taxiformis L1 and L2 in temperate and tropical areas. Despite the differences in predictions among modelling approaches, a number of regions were identified as zones with high invasion risk for A. taxiformis L2. Since range shifts are likely climate‐driven phenomena, future invasive behavior cannot be excluded for the rest of the lineages.     

Early Neolithic agriculture (2700–2000 <Emphasis Type="SmallCaps">bc</Emphasis>) and Kushan period developments (<Emphasis Type="SmallCaps">ad</Emphasis> 100–300): macrobotanical evidence from Kanispur in Kashmir,India

Here we report the results of an archaeobotanical study carried out on Ceramic Neolithic (2700–2000 bc) and Kushan period (ad 100–300) deposits at Kanispur, Kashmir, northern India. Botanical evidence from the Ceramic Neolithic phase reflects a nascent agriculture based on cold-tolerant crops (barley, wheats, lentil, field pea and grass pea) related to farming patterns of adjacent cultures and possibly evidence for the earliest cultivation in the valley. The Kushan period is characterized by a double-cropping pattern, suggesting a change in agricultural practices associated with a population recovery following a post-Neolithic decline. Finds of Juglans regia, and Prunus cf. amygdalus shells in ceramic Neolithic levels, and Vitis cf. vinifera, Emblica officinalis and Ziziphus cf. nummularia in addition to J. regia, and P. cf. amygdalus during Kushan times suggest that horticulture and foraging played an important role in the diet of the occupants. The present dataset provides new absolute dates for the subsistence economy of the Ceramic Neolithic population in the Kashmir Valley. We also present the oldest directly dated wheat and barley in Kashmir.     

Trends in sightings and population structure of white sharks, <Emphasis Type="Italic">Carcharodon carcharias,</Emphasis> at Seal Island,False Bay,South Africa,and the emigration of subadult female sharks approaching maturity

A long-term sightings and photographic identification (photo-ID) database documented the inter-annual and monthly trends in white shark (Carcharodon carcharias) sightings, sex ratios and life-history stages at Seal Island, False Bay, South Africa, over the period 2004–2012. A total of 1105 sightings were recorded during 171 scientific surveys incorporating 577 h of observation (annual mean 64.10 h, range 23.71–178.69 h). The mean annual sighting rate was 1.87 (range 0.90–3.19) sharks per hour and sighting rates declined over the nine-year study period. It is unknown whether the decline resulted from an actual population decline, or was due to changes in shark distribution or environmental conditions, but it highlights the need for ongoing monitoring at this site. The overall sex ratio was 1.0: 1.0: 2.3 for male: female: unsexed sharks, respectively. Of the 1105 sightings, 39% (n = 433) were photo-ID’d, representing 303 individual sharks (112: 111: 80, male: female: unsexed) and 130 resightings. Of the 303 photo-ID’d sharks, 71% were sighted in a single year only, indicative of transient behaviour. Of the 29% of sharks that were resighted in more than one year, 65% were resighted in the following year, indicating site fidelity to Seal Island, whereas 35% skipped one or more years between encounters, indicative of a temporary absence. The majority (60%) of the photo-ID’d sharks were immature, 32% were subadult and only 8% were mature. No young-of-the-year sharks and few adults were recorded, indicating that Seal Island is not an adult aggregation site, nor a pupping or nursery area, but rather is best described as a seasonal feeding ground. Large females were rarely resighted again after they approached length-at-maturity (≥450 cm TL), whereas some adult males were recorded consistently across years. The emigration of subadult female sharks approaching maturity from Seal Island, combined with the small number of mature sharks of both sexes reported from any South African location, indicate that adult aggregation sites, and thus areas of reproductive importance, still remain unknown.