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The shift in political landscape in Virginia from former Democratic Governor Ralph Northam to current Republican Governor Glen Youngkin influenced educational policies in the commonwealth. Waving the banner of parental rights, the Youngkin administration began targeting LGBTQIA2S+ students and educators through legislation and policies such as SB 656 and Model Policies on Ensuring Privacy, Dignity, and Respect for All Students and Parents in Virginia's Public Schools. To understand the influence on rural school districts, this study asked how rural secondary English Language Arts educators understand and respond to these anti-LGBTQIA2S+ policies as well as the ways in which those educators feel that those policies influence the climate of their classrooms. Using a qualitative questionnaire and semi-structured interviews, we sought input from secondary English Language Arts teachers in one entirely rural region of Virginia. This study produced four findings: differing understandings and implementation of policies, curricular control, effects on student-teacher relationships, and humanizing practices. We provide a call-to-action and recommendations for classroom educators. [ABSTRACT FROM AUTHOR]

Abstract The amount and patterns of phylodiversity in a community are often used to draw inferences about the local and historical factors affecting community assembly and can be used to prioritize communities and locations for conservation. Because measures of phylodiversity are based on the topology and branch lengths of phylogenetic trees, which are affected by the number and diversity of taxa in the tree, these analyses may be sensitive to changes in taxon sampling and tree reconstruction methods. To investigate the effects of taxon sampling and tree reconstruction methods on measures of phylodiversity, we investigated the community phylogenetics of the Ordway‐Swisher Biological Station (Florida), which is home to over 600 species of vascular plants. We studied the effects of (a) the number of taxa included in the regional phylogeny; (b) random versus targeted sampling of species to assemble the regional species pool; (c) including only species from specific clades rather than broad sampling; (d) using trees reconstructed directly for the taxa under study compared to trees pruned from a larger reconstructed tree; and (e) using phylograms compared to chronograms. We found that including more taxa in a study increases the likelihood of observing significantly nonrandom phylogenetic patterns. However, there were no consistent trends in the phylodiversity patterns based on random taxon sampling compared to targeted sampling, or within individual clades compared to the complete dataset. Using pruned and reconstructed phylogenies resulted in similar patterns of phylodiversity, while chronograms in some cases led to significantly different results from phylograms. The methods commonly used in community phylogenetic studies can significantly impact the results, potentially influencing both inferences of community assembly and conservation decisions. We highlight the need for both careful selection of methods in community phylogenetic studies and appropriate interpretation of results, depending on the specific questions to be addressed.

Autotransporter proteins are localised to the bacterial surface and promote colonisation of host epithelial surfaces. Here, the authors present the crystal structure of autotransporter UpaB and show evidence for distinct binding sites for glycosaminoglycans and host fibronectin.

Summary: Recent availability of biodiversity data resources has enabled an unprecedented ability to estimate phylogenetically based biodiversity metrics over broad scales. Such approaches elucidate ecological and evolutionary processes yielding a biota and help guide conservation efforts. However, the choice of appropriate phylogenetic resources and underlying input data uncertainties may affect interpretation. Here, we address how differences among phylogenetic source trees and levels of phylogenetic uncertainty affect these metrics and test existing hypotheses regarding geographic biodiversity patterns across the diverse vascular plant flora of Florida, US. Ecological niche models for 1,490 Florida species were combined with a “purpose-built” phylogenetic tree (phylogram and chronogram), as well as with trees derived from community resources (Phylomatic and Open Tree of Life). There were only modest differences in phylodiversity metrics given the phylogenetic source tree and taking into account the level of phylogenetic uncertainty; we identify similar areas of conservation interest across Florida regardless of the method used. : Spatial Phylogenetics; Plant Biology; Biogeography Subject Areas: Spatial Phylogenetics, Plant Biology, Biogeography

Phosphoinositides are members of a family of minor phospholipids that make up about 1% of all lipids in most cell types. Despite their low abundance they have been found to be essential regulators of neuronal activities such as action potential firing, release and re-uptake of neurotransmitters, and interaction of cytoskeletal proteins with the plasma membrane. Activation of several different neurotransmitter receptors can deplete phosphoinositide levels by more than 90% in seconds, thereby profoundly altering neuronal behavior; however, despite the physiological importance of this mechanism we still lack a profound quantitative understanding of the connection between phosphoinositide metabolism and neuronal activity. Here, we present a model that describes phosphoinositide metabolism and phosphoinositide-dependent action potential firing in sympathetic neurons. The model allows for a simulation of activation of muscarinic acetylcholine receptors and its effects on phosphoinositide levels and their regulation of action potential firing in these neurons. In this paper, we describe the characteristics of the model, its calibration to experimental data, and use the model to analyze how alterations of surface density of muscarinic acetylcholine receptors or altered activity levels of a key enzyme of phosphoinositide metabolism influence action potential firing of sympathetic neurons. In conclusion, the model provides a comprehensive framework describing the connection between muscarinic acetylcholine signaling, phosphoinositide metabolism, and action potential firing in sympathetic neurons which can be used to study the role of these signaling systems in health and disease.

Human zinc transporter 1 (hZnT1) belongs to the cation diffusion facilitator (CDF) family. It plays a major role in transporting zinc (Zn2+) from the cytoplasm across the plasma membrane and into the extracellular space thereby protecting cells from Zn2+ toxicity. Through homology with other CDF family members, ZnT1 is predicted to contain a transmembrane region and a soluble C-terminal domain though little is known about its biochemistry. Here, we demonstrate that human ZnT1 and a variant can be produced by heterologous expression in Saccharomyces cerevisiae cells and purified in the presence of detergent and cholesteryl hemisuccinate. We show that the purified hZnT1 variant has Zn2+/H+ antiporter activity. Furthermore, we expressed, purified and characterized the soluble C-terminal domain of hZnT1 (hZnT1-CTD) in a bacterial expression system. We found that the hZnT1-CTD melting temperature increases at acidic pH, thus, we used an acetate buffer at pH 4.5 for purifications and concentration of the protein up to 12 mg/mL. Small-angle X-ray scattering analysis of hZnT1-CTD is consistent with the formation of a dimer in solution with a V-shaped core.

Premise Commonly used molecular techniques such as next‐generation sequencing require reliable methods to extract DNA quickly and efficiently. Secondary compounds within plant tissues make this requirement all the more challenging, often forcing researchers to test different extraction methods tailored to their particular species of interest in order to obtain large amounts of high‐quality genomic DNA. The opportunities provided by high‐throughput, next‐generation sequencing only exacerbate these problems, especially when trying to extract DNA from multiple species at the same time. Several methods have attempted to resolve the challenges of obtaining suitable DNA from plants; however, a rapid, high‐yield, high‐quality, and highly scalable DNA extraction method is still needed. Methods and Results We present a rapid DNA extraction protocol that utilizes a buffer with relatively large amounts of cetyltrimethylammonium bromide (CTAB) and sodium chloride, combined with a silica maxi‐column cleanup of the extracted DNA. The new method is easy to implement using standard equipment and inexpensive reagents. The entire procedure (from grinding to the final elution) can be completed in less than two hours for a single sample and can be easily scaled to meet desired research goals. It works on diverse green plants with highly varied secondary chemistries (e.g., ferns, gymnosperms, and phylogenetically divergent angiosperms). Conclusions Application of the protocol to various plant species yielded DNA of high quality in less than two hours and can be adjusted to extract DNA at large (maxi‐preps) or small (96‐well minipreps) scales. We anticipate that our method will be of wide utility for rapidly isolating large quantities of quality genomic DNA from diverse plant species and will have broad applications in phylogenetic studies utilizing PCR and short‐read DNA sequencing.

This perspective describes advances in determining membrane protein structures in lipid bilayers using small-angle neutron scattering (SANS). Differentially labeled detergents with a homogeneous scattering length density facilitate contrast matching of detergent micelles; this has previously been used successfully to obtain the structures of membrane proteins. However, detergent micelles do not mimic the lipid bilayer environment of the cell membrane in vivo. Deuterated vesicles can be used to obtain the radius of gyration of membrane proteins, but protein-protein interference effects within the vesicles severely limits this method such that the protein structure cannot be modeled. We show herein that different membrane protein conformations can be distinguished within the lipid bilayer of the bicontinuous cubic phase using contrast-matching. Time-resolved studies performed using SANS illustrate the complex phase behavior in lyotropic liquid crystalline systems and emphasize the importance of this development. We believe that studying membrane protein structures and phase behavior in contrast-matched lipid bilayers will advance both biological and pharmaceutical applications of membrane-associated proteins, biosensors and food science.

In order to introduce the concepts of biocatalysis and its utility in synthesis to organic chemistry students, a multistep synthesis of endogenous cannabinergic ligand 2-arachidonoylglycerol (2-AG) was tailored for use as a laboratory exercise. Over four weeks, students successfully produced 2-AG, purifying and characterizing products at each stage of the reaction series. This synthetic series also reinforces the importance of executing reactions under inert atmosphere, the strategic placement of protecting groups, and regioselectivity.

Music interventions support functional outcomes, improve mood, and reduce symptoms of depression in neurorehabilitation. Neurologic music therapy (NMT) has been reported as feasible and helpful in stroke rehabilitation but is not commonly part of multidisciplinary services in acute or subacute settings. This study assessed the feasibility and acceptability of delivering NMT one-day-per-week in a subacute neurorehabilitation centre over 15 months. Data were collected on the number of referrals, who referred, sessions offered, attended, and declined, and reasons why. Staff, patients, and their relatives completed questionnaires rating the interventions. Patients completed the Visual Analog Mood Scales (VAMS) pre and post a single session. Forty-nine patients received 318 NMT sessions (83% of sessions offered). NMT was rated as helpful or very helpful as part of the multidisciplinary team (n = 36). The highest ratings were for concentration, arm and hand rehabilitation, and motivation and mood. VAMS scores (n = 24) showed a reduction in ‘confused’ (−8.6, p = 0.035, effect size 0.49) and an increase in ‘happy’ (6.5, p = 0.021, effect size = 0.12) post NMT. The data suggest that a one-day-per-week NMT post in subacute neurorehabilitation was feasible, acceptable, and helpful, supporting patient engagement in rehabilitation exercises, mood, and motivation.

By using a combination of experimental neutron scattering techniques, it is possible to obtain a statistical perspective on red blood cell (RBC) shape in suspensions, and the inter-relationship with protein interactions and dynamics inside the confinement of the cell membrane. In this study, we examined the ultrastructure of RBC and protein–protein interactions of haemoglobin (Hb) in them using ultra-small-angle neutron scattering and small-angle neutron scattering (SANS). In addition, we used the neutron backscattering method to access Hb motion on the ns time scale and Å length scale. Quasi-elastic neutron scattering (QENS) experiments were performed to measure diffusive motion of Hb in RBCs and in an RBC lysate. By using QENS, we probed both internal Hb dynamics and global protein diffusion, on the accessible time scale and length scale by QENS. Shape changes of RBCs and variation of intracellular Hb concentration were induced by addition of the Na+-selective ionophore monensin and the K+-selective one, valinomycin. The experimental SANS and QENS results are discussed within the framework of crowded protein solutions, where free motion of Hb is obstructed by mutual interactions.

Although fMRI studies have produced considerable evidence for differences in the spatial connectivity of resting-state brain networks in persons with autism spectrum disorder (ASD) relative to typically developing (TD) peers, little is known about the temporal dynamics of these brain networks in ASD. The aim of this study was to examine the EEG microstate architecture in children with ASD as compared to TD at rest in two separate conditions – eyes-closed (EC) and eyes-open (EO). EEG microstate analysis was performed on resting-state data of 13 ASD and 13 TD children matched on age, gender, and IQ. We found that children with ASD and TD peers produced topographically similar canonical microstates at rest. Group differences in the duration and frequency of these microstates were found primarily in the EC resting-state condition. In line with previous fMRI findings that have reported differences in spatial connectivity within the salience network (previously correlated with the activity of microstate C) in ASD, we found that the duration of activation of microstate C was increased, and the frequency of microstate C was decreased in ASD as compared to TD in EC resting-state. Functionally, these results may be reflective of alterations in interoceptive processes in ASD. These results suggest a unique pattern of EEG microstate architecture in ASD relative to TD during resting-states and also that EEG microstate parameters in ASD are susceptible to differences in resting-state conditions.

Most species of modern iguanas (Iguania, Iguanidae) dig burrows for dwelling and nesting, yet neither type of burrow has been interpreted as trace fossils in the geologic record. Here we describe and diagnose the first known fossil example of an iguana nesting burrow, preserved in the Grotto Beach Formation (Early Late Pleistocene, ~115 kya) on San Salvador Island, The Bahamas. The trace fossil, located directly below a protosol, is exposed in a vertical section of a cross-bedded oolitic eolianite. Abundant root traces, a probable land-crab burrow, and lack of ghost-crab burrows further indicate a vegetated inland dune as the paleoenvironmental setting. The trace fossil matches dimensions and overall forms of burrows made by modern iguanas, and internal structures indicate active backfilling consistent with modern iguana nesting burrows. The trace fossil is also located on an island with a modern native species of rock iguana (Cyclura riyeli riyeli), suggesting a presence of iguanas on San Salvador since the Late Pleistocene. This nesting burrow may provide a search image for more fossil iguana burrows in The Bahamas and other places with long-established iguana species and favorable geological conditions for preserving their burrows.

Background: The major causes of hepatocellular carcinomas are Aflatoxin, hepatitis B and hepatitis C viruses.Alpha tocopherol and its acetate and succinate esters have each been reported as counteracting cancer development in humans and rodents. We have investigated their salutary effect in both poor and high quality diets in rainbow trout Oncorhynchus mykiss as a model. Methods: Hepatocellular carcinomas (HCCs) were induced in rainbow trout by dietary aflatoxin B1 (AfB1). A matrix of different levels of several vitamins and vitamin analogues were included in selected diets as possible anticancer agents. Identification of HCCs was made by histopathology. Results: 1.) Elevated dietary tocopheryl acetate (E-Ac) caused a marked increase in liver size and in AfB1-induced HCCs in rainbow trout. 2.) Poor diets increased the HCC incidence. 3.) Elevated dietary tocopheryl succinate (E-Su) nearly eliminated HCC development in fish fed complete diets. Tocopheryl succinate in poor diets reduced HCCs by 77% compared to tocopheryl acetate diets. 4.) Trans-retinoic acid also reduced HCC incidence. 5.) Vitamins A and D deficiency caused tumor increases but had no effect on liver size. 6.) The use of casein and dextrin in the place of soybean textured vegetable protein, in poor diets nearly eliminated the HCC risk. 7.) Trout sera showed all three vitamin forms; free α-tocopherol (E-OH), tocopheryl acetate (E-Ac) and tocopheryl succinate (E-Su), from diets containing any of these vitamin analogues, suggesting both de-esterification and trans-esterification. 8.) E-Su is discussed in the light of an anti-cancer agent that is non toxic to normal tissue but that cohorts to it are needed. Conclusions: Increased dietary E-Ac escalated AfB1 induced HCCs and caused hepatomegaly in rainbow trout, while E-Su eliminated the HCC risk as shown by histopathology. Keywords: Aflatoxin, Tocopheryl-acetate, Tocopheryl-succinate, Hepatocellular, carcinomas, Hepatomegaly

Polly Heine,1 Laura Sellar,1 Sue Whitten,2 Priti Bajaj2 1Fertility Team, The Agora Gyneacology and Fertility Centre, Hove, 2Medical Affairs, Ferring Pharmaceuticals, West Drayton, UK Purpose: The aim of this audit was to assess the overall experience and patient convenience of vaginal progesterone tablets (Lutigest®, marketed as Endometrin® in the USA) used for luteal phase support (LPS) during in vitro fertilization (IVF) treatment.Patients and methods: This questionnaire-based audit included responses from 100 patients undergoing IVF treatment at six IVF clinics in the UK from September 2015 to November 2016. Fourteen days after starting progesterone supplementation for LPS during their IVF treatment, patients rated overall experience and perceived convenience of the prescribed progesterone by completing a questionnaire.Results: Of the 100 patients included, 96 received vaginal progesterone tablets for LPS. Overall, 53.1% (51/96) indicated that the progesterone tablets were “very easy” to use; 42.7% (41/96) and 44.8% (43/96) found it “very convenient” or “neither convenient or inconvenient” to administer the tablet, respectively. Overall experience with using progesterone tablets was rated as “very comfortable” by 34.4% (33/96) and “neither comfortable or uncomfortable” by 56.3% (54/96) of patients. The applicator was used by 93.8% (90/96) of patients to administer the tablet, and 86.5% (83/96) indicated that the applicator was easy to clean for repeated use. A total of 33 patients had a previous IVF cycle during which they were prescribed vaginal progesterone pessaries for LPS. Compared with progesterone pessaries, the majority found treatment with progesterone tablets to be more comfortable (60.6%; 20/33) and more convenient (57.6%; 19/33) and indicated that the progesterone tablet was their preferred progesterone formulation for LPS (60.6%; 20/33).Conclusion: These findings offer insights into real-world patient experiences with the progesterone vaginal tablet formulation. The results suggest overall patient convenience, ease, and comfort with using progesterone vaginal tablets for LPS. The majority of patients found progesterone vaginal tablets more convenient and comfortable to use compared with progesterone pessaries. Keywords: vaginal progesterone tablets, progesterone pessary, progesterone gel, luteal phase support, in vitro fertilization, patient experience, convenience

The social change model has proven an effective and widely utilized model assisting college students in leadership development toward positive social change. However, while this particular model gives much needed attention to the process of development leading to social change, it fails to acknowledge the external factors which significantly affect the student's capacity to develop effectively. This article discusses these issues and proposes "comprehensive support" as an additional component of the model, including two additional "C's." This addition attempts to more fully acknowledge the external realities affecting the potential for effective student leadership among college students.

Women have comprised the majority of accounting graduates with bachelor and master degrees since the 1980s and 1990s, respectively, but currently only account for about 30% of doctoral students. Their representation in the accounting professoriate is essentially the same. Supportive strategies, such as mentoring, assist disciplines in which women are under-represented in attracting and retaining them in the work environment. Work engagement is one measure of a person's commitment to their current employer. Therefore, understanding which aspects of mentoring increase the level of work engagement of those women currently in academia is important and can guide institutions in developing mentoring programs. In my study, I investigated factors that predict work engagement for female accounting professors.

Bacterial disulfide isomerases shuffle incorrect disulfide bonds. Here, the authors structurally characterize the disulfide isomerase ScsC fromProteus mirabilisand identify a functionally important shape-shifting motif that allows ScsC to adopt a diverse range of conformations and enable swarming in the presence of copper stress.

Uniting diverse stakeholders through communication, education or building a collaborative 'common vision' for biodiversity management is a recommended approach for enabling effective conservation in regions with multiple uses. However, socially focused strategies such as building a collaborative vision can require sharing scarce resources (time and financial resources) with the on-ground management actions needed to achieve conservation outcomes. Here we adapt current prioritisation tools to predict the likely return on the financial investment of building a stakeholder-led vision along with a portfolio of on-ground management strategies. Our approach brings together and analyses expert knowledge to estimate the cost-effectiveness of a common vision strategy and on-ground management strategies, before any investments in these strategies are made. We test our approach in an intensively-used Australian biodiversity hotspot with 179 threatened or at-risk species. Experts predicted that an effective stakeholder vision for the region would have a relatively low cost and would significantly increase the feasibility of on-ground management strategies. As a result, our analysis indicates that a common vision is likely to be a cost-effective investment, increasing the expected persistence of threatened species in the region by 9 to 52%, depending upon the strategies implemented. Our approach can provide the maximum budget that is worth investing in building a common vision or another socially focused strategy for building support for on-ground conservation actions. The approach can assist with decisions about whether and how to allocate scarce resources amongst social and ecological actions for biodiversity conservation in other regions worldwide.

The α and polyproline II (PPII) basins are the two most populated regions of the Ramachandran map when constructed from the protein coil library, a widely used denatured state model built from the segments of irregular structure found in the Protein Data Bank. This indicates the α and PPII conformations are dominant components of the ensembles of denatured structures that exist in solution for biological proteins, an observation supported in part by structural studies of short, and thus unfolded, peptides. Although intrinsic conformational propensities have been determined experimentally for the common amino acids in short peptides, and estimated from surveys of the protein coil library, the ability of these intrinsic conformational propensities to quantitatively reproduce structural behavior in intrinsically disordered proteins (IDPs), an increasingly important class of proteins in cell function, has thus far proven elusive to establish. Recently, we demonstrated that the sequence dependence of the mean hydrodynamic size of IDPs in water and the impact of heat on the coil dimensions, provide access to both the sequence dependence and thermodynamic energies that are associated with biases for the α and PPII backbone conformations. Here, we compare results from peptide-based studies of intrinsic conformational propensities and surveys of the protein coil library to those of the sequence-based analysis of heat effects on IDP hydrodynamic size, showing that a common structural and thermodynamic description of the protein denatured state is obtained.