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Ecological restoration is increasingly employed to restore degraded or destroyed ecosystems and evaluation of restoration success requires that natural community dynamics be understood. Wet prairies in the Southeast US have diverse plant communities subject to disturbances including fire, drought, flooding, tropical storms, and freezes. This habitat covers a fraction of its former range and reversing that trend requires ecological restoration; but, long-term data on the dynamics of this system are rare. We analyzed a 12-year plant community composition dataset from a fire-maintained Gulf of Mexico Coastal Plain wet prairie to characterize plant community dynamics and identify indicator species. The site was compared to a nearby wetland mitigation project attempting to restore a wet prairie that had been converted into slash pine (Pinus elliottii) plantation. The reference site had higher species richness, was dominated by Aristida spp., and was stable despite extreme climatic conditions. The restoration site transitioned from dense pine and understory canopy to an open, grassy community, but did not follow a trajectory toward the reference state. Restoration efforts were impeded by woody debris, accumulated duff, soil disturbance from logging, and storm surge from hurricanes. Continued application of prescribed fire and decomposition activity should remove the organic matter and promote establishment of wet prairie species, but on a longer time scale than expected. Although the pine plantation did not return to a typical wet prairie during the study period, the record of community dynamics of the reference wet prairie provides information on the resilience of the habitat type under prescribed fire management, and serves as a reference for restoration efforts and management of this threatened habitat.

We examined the static and dynamic characters of the red luminescence in the protein-Au(III) compounds, directly comparing multiple proteins: BSA, OVA, trypsin, and insulin. These four protein-Au(III) complexes showed a nearly identical excitation-emission pattern, not only the wavelength of luminescence (λem ∼ 640 nm). Lifetimes of the red luminescence shared a common value of ∼300 ns. Kinetics of the luminophore formation was consistently described by a Langmuir-type chemisorption of Au(III) for these proteins, coinciding with the protein conformation change at pH ∼ 10. These observations and the protein structural analyses support that the red luminophore formation involves Au(III) coordination to a common motif within these proteins.

BACKGROUND AND OBJECTIVES The aim of this study was to evaluate the role of T- and B-regulatory cells (Tregs and Bregs) in the pathogenesis of idiopathic granulomatous mastitis (IGM). METHODS This study includes 47 patients with pathologically proven IGM (Group P) and 26 healthy subjects (Group C). The patients in Group P were divided into two groups according to whether their lesions were active (Group PA, n: 21) or in remission (Group PR, n: 26). By using flow-cytometry, the frequencies of CD3+CD4+CD45RA-Foxp3high activated Tregs (aTregs), CD3+CD4+CD45RA-Foxp3low non-suppressive Tregs, CD3+CD4+CD45RA+Foxp3low resting Tregs (rTregs), CD3+CD4+CD25+Foxp3- T-effector cells (Teff), total Tregs and Bregs were analyzed in all subjects. RESULTS The frequency of the Teff cells was statistically higher in Group P when compared with Group C (p =.004). The Foxp3 expression of Treg cells and the frequency of non- suppressive Tregs in Group P were statistically lower than Group C (p =.032 and p =.02, respectively). In addition, Group PR's Foxp3 expressions were statistically lower than Group C (p =.027); Group PR's aTregs ratio was statistically lower than Group PA (p =.021); and the non-suppressive Tregs ratio of Group PR was lower than both Group PA and Group C (p =.006 and p

The spatial dissemination of three prevalent taxa of sooty blotch and flyspeck (SBFS) fungi under several levels of precipitation was compared during 2015 and 2016 in an Iowa apple orchard. Overhead irrigation was used to supplement ambient precipitation in order to insure SBFS spore dissemination and colony development. There were five irrigation levels, involving 1-min-long periods of irrigation that were imposed either once or twice per hour at intervals of 3, 6, or 12 h, as well as a nonirrigated control. Preselected apple fruit were inoculated with one of the three SBFS taxa to serve as sources of inoculum. Dissemination from these inoculated apple fruit was assessed at harvest by counting SBFS colonies on water-sprayed and nontreated fruit. As a further control, additional fruit were enclosed in fruit bags throughout the fruit development period. In both 2015 and 2016, the number of colonies of the SBFS fungus Peltaster gemmifer per apple increased sharply as the duration of irrigation increased, whereas the number of colonies of Microcyclosporella mali increased to a lesser extent and Stomiopeltis sp. RS1 showed no increase. In 2015, the linear relationship between the duration of irrigation-imposed precipitation levels and the number of colonies on the water-sprayed apple fruit was similar for P. gemmifer (slope = 0.09), Stomiopeltis sp. RS1 (slope = 0.07), and Microcyclosporella mali (slope = 0.13); whereas, in 2016, the slope was higher for P. gemmifer (0.28) than for Stomiopeltis sp. RS1 (−0.09) or M. mali (0.06). The results indicated that dissemination of P. gemmifer increased sharply in response to increased irrigation-imposed precipitation, and that dissemination patterns differed considerably among the three SBFS taxa. The apparent advantage of P. gemmifer in precipitation-triggered dissemination may stem from its ability to produce spores rapidly by budding. To our knowledge, this is the first article to assess splash dispersal by SBFS fungi in the field and the first to document taxon-specific patterns of dissemination in this pathogen complex.

Summary Background Very low birthweight or preterm infants are at increased risk of adverse outcomes including sepsis, necrotising enterocolitis, and death. We assessed whether supplementing the enteral diet of very low-birthweight infants with lactoferrin, an antimicrobial protein, reduces all-cause mortality or major morbidity. Methods We did a multicentre, double-blind, pragmatic, randomised superiority trial in 14 Australian and two New Zealand neonatal intensive care units. Infants born weighing less than 1500 g and aged less than 8 days, were eligible and randomly assigned (1:1) using minimising web-based randomisation to receive once daily 200 mg/kg pasteurised bovine lactoferrin supplements or no lactoferrin supplement added to breast or formula milk until 34 weeks' post-menstrual age (or for 2 weeks, if longer), or until discharge from the study hospital if that occurred first. Designated nurses preparing the daily feeds were not masked to group assignment, but other nurses, doctors, parents, caregivers, and investigators were unaware. The primary outcome was survival to hospital discharge or major morbidity (defined as brain injury, necrotising enterocolitis, late-onset sepsis at 36 weeks' post-menstrual age, or retinopathy treated before discharge) assessed in the intention-to-treat population. Safety analyses were by treatment received. We also did a prespecified, PRISMA-compliant meta-analysis, which included this study and other relevant randomised controlled trials, to estimate more precisely the effects of lactoferrin supplementation on late-onset sepsis, necrotising enterocolitis, and survival. This trial is registered with the Australian and New Zealand Clinical Trials Registry, ACTRN12611000247976. Findings Between June 27, 2014, and Sept 1, 2017, we recruited 1542 infants; 771 were assigned to the intervention group and 771 to the control group. One infant who had consent withdrawn before beginning lactoferrin treatment was excluded from analysis. In-hospital death or major morbidity occurred in 162 (21%) of 770 infants in the intervention group and in 170 (22%) of 771 infants in the control group (relative risk [RR] 0·95, 95% CI 0·79–1·14; p=0·60). Three suspected unexpected serious adverse reactions occurred; two in the lactoferrin group, namely unexplained late jaundice and inspissated milk syndrome, but were not attributed to the intervention and one in the control group had fatal inspissated milk syndrome. Our meta-analysis identified 13 trials completed before Feb 18, 2020, including this Article, in 5609 preterm infants. Lactoferrin supplements significantly reduced late-onset sepsis (RR 0·79, 95% CI 0·71–0·88; p Interpretation Lactoferrin supplementation did not improve death or major morbidity in this trial, but might reduce late-onset sepsis, as found in our meta-analysis of over 5000 infants. Future collaborative studies should use products with demonstrated biological activity, be large enough to detect moderate and clinically important effects reliably, and assess greater doses of lactoferrin in infants at increased risk, such as those not exclusively receiving breastmilk or infants of extremely low birthweight. Funding Australian National Health and Medical Research Council.

Developing effective butterfly monitoring strategies is key to understanding how butterflies interact with urban environments, and, in turn, to developing local conservation practices. We investigated two urban habitat types (public gardens and restored/reconstructed prairies) and compared three survey methods (Pollard transects, purposive point counts, and random point counts) to determine which was most productive for detecting butterflies and assessing family diversity. We conducted 66 butterfly surveys by using each method (198 total) from May through September in 2015 and 2016 at six sites (three public gardens and three prairies) in Ames, Ankeny and Des Moines, Iowa. All survey methods were used on 11 sampling dates at each site. Overall, we observed 2,227 butterflies representing 38 species: 1,076 in public gardens and 1,151 in prairie areas. We used a smaller data set standardized for survey effort, including 1,361 of these sightings, to compare survey methods and habitat types. Although there were no significant differences in number of butterfly sightings between the two habitats, more sightings (798) were documented by using purposive point counts when compared to Pollard transects (297) or random point counts (266) (for both comparisons, p

The activation mechanism of the ErbB family of receptors is of considerable medical interest as they are linked to a number of human cancers, including an aggressive form of breast cancer. In the rat analogue of the human ErbB2 receptor, referred to as Neu, a point mutation in the transmembrane domain (V664E) has been shown to trigger oncogenic transformation. While the structural impact of this mutation has been widely studied in the past to yield models for the active state of the Neu receptor, little is known about the impact of cholesterol on its structure. Given previous reports of the influence of cholesterol on other receptor tyrosine kinases (RTKs), as well as the modulation of lipid composition in cancer cells, we wished to investigate how cholesterol content impacts the structure of the Neu transmembrane domain. We utilized high-resolution magic angle spinning solid-state NMR to measure 13C–13C coupling of selectively labelled probe residues in the Neu transmembrane domain in lipid bilayers containing cholesterol. We observe inter-helical coupling between residues that support helix–helix interactions on both dimerization motifs reported in the literature (A661-XXX-G665 and I659-XXX-V663). We further explore how changes in cholesterol concentration alter transmembrane domain interactions and the properties and mechanics of the bilayer. We interpret our results in light of previous studies relating RTK activity to cholesterol enrichment and/or depletion, and propose a novel model to explain our data that includes the recognition and binding of cholesterol by the Neu transmembrane domain through a putative cholesterol-recognition/interaction amino acid consensus sequence.\ud \ud

Much field research on perennial bioenergy crops confounds effects of plant age with those of the growing season, which increases uncertainty and the potential for erroneous conclusions, particularly in maturing stands. Most studies rely on stands planted in a single year and measured across multiple subsequent seasons. These “single-start” designs lack statistical power to separate temporal from environment effects. We used a staggered start experimental design to learn if increased statistical power clarified understanding of Miscanthus × giganteus nitrogen (N) needs. We conducted a staggered start experiment with three planting years and five N rates during the M. × giganteus yield-building phase at three sites across IA, USA. Third-year yields were 21.0, 25.0, and 27.1 Mg dry matter (DM) ha−1 at the northwest (NW), central, and southeast (SE) sites, respectively. Nitrogen fertilization effects changed with establishment conditions, but not with plant age. At the most N responsive site, N fertilizer changed yields at all stand ages, but not in every year. Yield increases of 150%, 36%, and 40% were observed in 1-, 2-, and 3-year-old stands, respectively, with N addition. Nitrogen effects on 1-year-old stands were positive in SE IA (2.7 kg DM kg−1 N added), negative (− 2.3 kg DM kg−1 N) in NW IA, and variable in central IA (− 2.2–9.6 kg DM kg−1 N), suggesting a site–year-specific response. Yield increases between the first and second years varied by > 100% depending on establishment conditions, highlighting the need for repeated planting before determining economic and agronomic crop viability.

We revisit the prevailing hypothesis that the red fluorophore (λem = 640 nm) in the bovine serum albumin (BSA)–gold (Au) compound is a Au25 nanocluster. To examine the hypothesis, we investigated t...

Antibiotic resistance is a significant threat to human health, with natural products remaining the best source for new antimicrobial compounds. Antimicrobial peptides (AMPs) are natural products with great potential for clinical use as they are small, amenable to customization, and show broad‐spectrum activities. Lynronne‐1 is a promising AMP identified in the rumen microbiome that shows broad‐spectrum activity against pathogens such as methicillin‐resistant Staphylococcus aureus and Acinetobacter baumannii. Here we investigated the structure of Lynronne‐1 using solution NMR spectroscopy and identified a 13‐residue amphipathic helix containing all six cationic residues. We used biophysical approaches to observe folding, membrane partitioning and membrane lysis selective to the presence of anionic lipids. We translated our understanding of Lynronne‐1 structure to design peptides which varied in the size of their hydrophobic helical face. These peptides displayed the predicted continuum of membrane‐lysis activities in vitro and in vivo, and yielded a new AMP with 4‐fold improved activity against A. baumannii and 32‐fold improved activity against S. aureus., Here we describe the biophysical characterization of Lynronne‐1, an antimicrobial peptide found in the bovine rumen microbiome. We reveal the location of an amphipathic helix containing all six cationic residues, and observe lipid‐selective folding, membrane‐binding and lysis. Our structural data were used to design three variants with changes in the size of the hydrophobic helical face, one of which yielded 32‐fold improved activity in vivo.

The reticulon family of integral membrane proteins are conserved across all eukaryotes and typically localize to the endoplasmic reticulum (ER), where they are involved in generating highly-curved tubules. We recently demonstrated that Reticulon-like protein B13 (RTNLB13) from Arabidopsis thaliana contains a curvature-responsive amphipathic helix (APH) important for the proteins’ ability to induce curvature in the ER membrane, but incapable of generating curvature by itself. We suggested it acts as a feedback element, only folding/binding once a sufficient degree of curvature has been achieved, and stabilizes curvature without disrupting the bilayer. However, it remains unclear whether this is unique to RTNLB13 or is conserved across all reticulons—to date, experimental evidence has only been reported for two reticulons. Here we used biophysical methods to characterize a minimal library of putative APH peptides from across the 21 A. thaliana isoforms. We found that reticulons with the closest evolutionary relationship to RTNLB13 contain curvature-sensing APHs in the same location with sequence conservation. Our data reveal that a more distantly-related branch of reticulons developed a ~ 20-residue linker between the transmembrane domain and APH. This may facilitate functional flexibility as previous studies have linked these isoforms not only to ER remodeling but other cellular activities.

Ketogenic diets have the potential to lower glucose availability to cancer cells. However, the effect that the resulting increase in ketone bodies has on cancer cells is not fully understood. The present study explored the effect of β-hydroxybutyrate (BHB) on glucose-deprived MCF-7 and T47D breast cancer cells. Cell proliferation was decreased in response to lower glucose conditions, which could not be rescued consistently by 10 or 25 mM BHB supplementation. In addition, gene expression levels were altered when cells were glucose deprived. Reducing glucose availability of cancer cells to 225 mg/l for 4 days significantly decreased the expression of 113 genes and increased the expression of 100 genes in MCF-7 breast cancer cells, and significantly decreased the expression of 425 genes and increased the expression of 447 genes in T47D breast cancer cells. Pathway enrichment analysis demonstrated that glucose deprivation decreased activity of the Hippo-Yap cell signaling pathway in MCF-7 breast cancer cells, whereas it increased the expression of genes in the NRF2-pathaway and genes regulating ferroptosis in T47D breast cancer cells. Treatment of glucose-deprived cells with 10 or 25 mM BHB significantly changed the expression of 14 genes in MCF-7 breast cancer cells and 40 genes in T47D breast cancer cells. No significant pathway enrichment was detected when glucose-deprived cells were treated with BHB. Both cell lines expressed the enzymes (OXCT1/2, BDH1 and ACAT1/2) responsible for metabolizing BHB to acetyl-CoA, yet expression of these enzymes was not altered by either glucose deprivation or BHB treatment. In the publicly available The Cancer Genome Atlas (TCGA), increased expression of ketone body-catabolizing enzymes was observed in various types of cancer based on mRNA expression z-scores. Increased expression of BDH1 and ACAT1 significantly decreased overall survival of patients with breast cancer in TCGA studies, while decreased OXCT1 expression non-significantly decreased overall survival. In conclusion, neither MCF-7 nor T47D breast cancer cells were affected by BHB during glucose deprivation; however, screening of tumors for activation of ketone body-metabolizing enzymes may be able to identify patients that will benefit from ketogenic diet interventions.

The geometric morphometrics method (GMM) is a technique to study scale and shape relationships of structures using Cartesian geometric coordinates rather than linear, areal (of area), or volumetric variables. The GMM has been shown to be of great value in many biological studies, but does not appear to have been used to examine equine skulls. In this exploratory study, 29 normal equine heads of three different age groups: 16 years old (N=10) were examined using the GMM technique. Computed tomography (CT) bone window DICOM images of equine skulls were reconstructed into isosurfaces (3-dimensional contoured surfaces), onto which anatomical landmarks were added using Stratovan Checkpoint® software. Data from 29 different landmarks were analysed using MorphoJ analysis, which applies a Procrustes fit, prior to reducing data dimensionality through principal component (PC) analysis. PCs with and without allometry (shape variation that can be explained by size changes) were considered. Allometric shape described by PC1 analysis accounted for 27% of variance. Loading pertaining to the following skull landmarks: the pterygoid process, bilaterally; caudal aspect of hard palate; tip of nasal bone; ethmoid sinuses, bilaterally; caudal aspect of the ventral conchal bulla, bilaterally and caudal aspect of the vomer bone suggest that these anatomical structures are predictive of age group. When allometric effects were removed, PC1 analysis was unable to distinguish horses by age group. Allometric shape differences could distinguish the youngest versus the two older age groups. The potential applications of GMM in equine diagnostic imaging are wide ranging and include the investigation of genetic-related changes in the equine skull shape and a more complete characterisation of the many conformation-related diseases affecting the teeth, jaws and sinonasal compartments of horses.

Superoxide dismutase (SOD) is known to be protective against oxidative stress-mediated skin dysfunction. Here we explore the potential therapeutic activities of RM191A, a novel SOD mimetic, on skin. RM191A is a water soluble, dimeric copper (Cu2+-Cu3+)-centred polyglycine coordination complex. It displays 10-fold higher superoxide quenching activity compared to SOD as well as significant anti-inflammatory activity through beneficial modulation of several significant inflammatory pathways in cells.We tested the therapeutic potential of RM191A in a topical gel using a human skin explant model and observed that it significantly inhibits UV-induced DNA damage in the epidermis and dermis, including cyclobutane pyrimidine dimers (CPD), 8-oxo-guanine (8-oxoG) and 8-nitroguanine (8NGO). RM191A topical gel is found to be safe and non-toxic in mice following month-long daily dosing at 0.19 mL/kg body weight. Moreover, it significantly accelerates excisional wound healing, and reduces 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin inflammation in mice.HighlightsSuperoxide dismutase mimetic RM191A is a highly stable copper (Cu2+-Cu3+)-polyglycine coordination complexRM191A exhibits potent antioxidant (10-fold more than that of superoxide dismutase) properties in vitroRM191A exhibits potent anti-inflammatory properties in vitro and in vivoRM191A protects human skin explants against UV-induced oxidative stress and DNA damageRM191A is non-toxic and readily bioavailable in miceRM191A attenuates TPA-induced skin inflammation and improves wound healing in mice

Superoxide dismutase (SOD) is known to be protective against oxidative stress-mediated skin dysfunction. Here we explore the potential therapeutic activities of RM191A, a novel SOD mimetic, on skin. RM191A is a water-soluble dimeric copper (Cu2+-Cu3+)-centred polyglycine coordination complex. It displays 10-fold higher superoxide quenching activity compared to SOD as well as significant antioxidant, anti-inflammatory and immunomodulatory activities through beneficial modulation of several significant inflammatory cytokines in vitro and in vivo. We tested the therapeutic potential of RM191A in a topical gel using a human skin explant model and observed that it significantly inhibits UV-induced DNA damage in the epidermis and dermis, including cyclobutane pyrimidine dimers (CPD), 8-oxo-guanine (8-oxoG) and 8-nitroguanine (8NGO). RM191A topical gel is found to be non-toxic, non-teratogenic and readily distributed in the body of mice. Moreover, it significantly accelerates excisional wound healing, reduces 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation and attenuates age-associated oxidative stress in skin, demonstrating both skin regenerative and geroprotective properties of RM191A., Graphical abstract Image 1, Highlights • RM191A is a Cu3+ containing coordination complex with 10-fold higher superoxide quenching activity compared to superoxide dismutase. • RM191A exhibits potent antioxidant, anti-inflammatory and immunomodulatory properties in vitro and in vivo. • RM191A protects human skin explants against UV-induced oxidative stress and DNA damage. • RM191A is non-toxic, non-teratogenic and readily bioavailable in mice. • RM191A promotes wound healing, and attenuates TPA-induced inflammation as well as age-associated oxidative stress in mouse skin.

Serum albumin-gold complexes exhibit UV-excitable red luminescence (λem = 640 nm) with unusual Stokes shifts compared with the innate UV/blue fluorescence arising from the aromatic residues. In order to understand the mechanism of this luminescence, we employed limited proteolysis and molecular cloning techniques and assessed the domain containing the red luminophore in bovine serum albumin (BSA) and human serum albumin (HSA). We identified that the luminophore is localized in a domain of serum albumin, residing within the N-terminus half.

The glycopeptide antibiotic vancomycin has been widely used to treat infections of Gram-positive bacteria including Clostridium difficile and methicillin-resistant Staphylococcus aureus. However, since its introduction, high level vancomycin resistance has emerged. The genes responsible require the action of the two-component regulatory system VanSR to induce expression of resistance genes. The mechanism of detection of vancomycin by this two-component system has yet to be elucidated. Diverging evidence in the literature supports activation models in which the VanS protein binds either vancomycin, or Lipid II, to induce resistance. Here we investigated the interaction between vancomycin and VanS from Streptomyces coelicolor (VanSSC), a model Actinomycete. We demonstrate a direct interaction between vancomycin and purified VanSSC, and traced these interactions to the extracellular region of the protein, which we reveal adopts a predominantly α-helical conformation. The VanSSC-binding epitope within vancomycin was mapped to the N-terminus of the peptide chain, distinct from the binding site for Lipid II. In targeting a separate site on vancomycin, the effective VanS ligand concentration includes both free and lipid-bound molecules, facilitating VanS activation. This is the first molecular description of the VanS binding site within vancomycin, and could direct engineering of future therapeutics.

Hollow-bearing trees provide critical habitat resources for forest fauna, yet there is evidence of a ubiquitous decline in the large, old trees most likely to provide this resource. Fire can influence the formation and persistence of tree hollows. In this study, we investigated the effects of stand-level fire history and individual tree attributes on tree hollow abundance in two forest types in Namadgi National Park in the Australian Capital Territory: subalpine woodlands dominated by Eucalyptus pauciflora Sieber ex Spreng (snow gum); and tall-open E. delegatensis R.T.Baker (alpine ash) montane forest. These forests can be differentiated by their distinct response to fire; E. pauciflora resprouts following fire and E. delegatensis reproduces exclusively via seed. We employed a ground-based approach to measure 1044 trees across 36 sites selected by forest type and fire history as recorded since 1920. For both species, hollow abundance decreased with total fire count at stand level and increased for E. delegatensis in response to an extensive wildfire that occurred in the study area in 2003. The probability of a tree containing a hollow increased with tree diameter and if the tree was dead. Our results show that fire frequency and severity have strong implications for tree hollow abundance in montane and subalpine eucalypt forests.

Novel single nucleotide polymorphism loci were identified for teak (Tectona grandis Linn. f.) using a combined Next Generation Sequencing approach on the Illumina MiSeq system with genotyping on the Agena MassARRAY iPLEX™ platform. Three hundred and forty four putative loci were identified through sequencing of DNA fragments following a modified double digest Restriction Associated DNA Sequencing method. One hundred and eighty nine loci successfully amplified in 172 individuals from 20 populations in Myanmar. A total of 33 loci were removed from the final panel as two showed evidence of deviation from Hardy–Weinberg equilibrium, 17 were significantly linked and 23 were non-polymorphic. For the remaining 156 loci, expected heterozygosity ranged from 0.041 to 0.456 and FST from 0.062 to 0.632. These genetic resources will prove useful for future studies into the population genetics and phylogeography of this important and iconic timber species.

Ehrlichioses and anaplasmosis have undergone dramatic increases in incidence, and the geographic ranges of their occurrence and vectors have also expanded. There is marked underreporting of these diseases owing to deficient physician awareness and knowledge of the illnesses as well as limited access to appropriate diagnostic tests. Human monocytic ehrlichiosis and anaplasmosis are life threatening diseases with estimated case fatality rates of 2.7 and 0.3%, respectively. However, knowledge of their full range of signs and symptoms is incomplete, and the incidence of subclinical infections is unknown. Currently available laboratory diagnostic methods are poorly utilized, and with the exception of nucleic acid amplification tests are not useful for diagnosis during the acute stage of illness when timely treatment is needed. The Ehrlichiosis and Anaplasmosis Subcommittee of the Tick-Borne Disease Working Group recommended active clinical surveillance to determine the true incidence, full clinical spectrum, and risk factors for severe illness, as well as standardized surveillance of ticks for these pathogens, and enhanced education of primary medical caregivers and the public regarding these diseases. The subcommittee identified the needs to develop sensitive, specific acute stage diagnostic tests for local clinical laboratories and point-of-care testing, to develop approaches for utilizing electronic medical records, data mining, and artificial intelligence for assisting early diagnosis and treatment, and to develop adjunctive therapies for severe disease.

The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), vectors the bacterium causing citrus greening disease, which has devastated citrus production worldwide wherever it has been introduced. To help monitor and target D. citri populations in commercial groves, thereby facilitating more effective management of citrus greening disease, a prototype device has been developed that mimics D. citri female vibrational communication signals, attracting males to a trap. For this report, effects of the device on male D. citri searching behavior were assessed to consider potential improvements in field applications. Forty-five percent of the males that searched towards the female signal mimic reached the source. In addition, the mean latencies before the initiation of calling and searching responses by males that reached the source were significantly lower than for those that missed, which suggests that trapping efficiency is strongly influenced by variability in male responsiveness to searching cues. Consequently, it is likely that the trapping efficiency of vibration traps could be increased further if they were modified to make use of additional cues strongly attractive to males, such as citrus flush olfactory and visual cues. ResumenEl silido asiatico de los citricos Diaphorina citri Kuwayama (Hemiptera: Liviidae) es un vector de la bacteria causante de la enfermedad del enverdecimiento de los citricos, que ha devastado la produccion de citricos en todo el mundo dondequiera que este ha sido introducido. Para ayudar a monitorear y enfocarse a las poblaciones de D. citri en los bosques comerciales, facilitando asi un manejo mas efectivo de la enfermedad del enverdecimiento de los citricos, se ha desarrollado un aparato prototipo que imita las senales vibracionales de comunicacion de las hembras de D. citri, que atraen a machos a las trampas. Para este informe, se evaluaron los efectos del aparato sobre el comportamiento de busqueda de los machos de D. citri para considerar posibles mejoras en las aplicaciones de campo. El 45% de los machos que buscaban hacia la senal que imitaba la hembra llegaron a la fuente. Ademas, el promedio de la latencia antes de la iniciacion de la llamada y las respuestas de busqueda por los machos que llegarona la fuente fueron significativamente mas bajos que para los que se fallaron, lo que sugiere que la eficiencia de captura esta fuertemente influenciada por la variabilidad en la capacidad de respuesta de los machos a buscar senales. En consecuencia, es probable que la eficacia de captura de trampas de vibracion podria aumentar aun mas si se modificaron para hacer uso de senales adicionales fuertemente atractivas para los machos, tales como senales olfativas y visuales de los brotes de nuevas hojas en citricos.View this article in BioOne

A warning system for the sooty blotch and flyspeck (SBFS) fungal disease complex of apple, developed originally for use in the southeastern United States, was modified to provide more reliable assessment of SBFS risk in Iowa. Modeling results based on previous research in Iowa and Wisconsin had suggested replacing leaf wetness duration with cumulative hours of relative humidity (RH) ≥97% as the weather input to the SBFS warning system. The purpose of the present study was to evaluate the performance of a RH-based SBFS warning system, and to assess the potential economic benefits for its use in Iowa. The warning system was evaluated in two separate sets of trials—trial 1 during 2010 and 2011, and trial 2 during 2013–2015—using action thresholds based on cumulative hours of RH ≥97% and ≥90%, respectively, in conjunction with two different fungicide regimes. The warning system was compared with a traditional calendar-based system that specified spraying at predetermined intervals of 10 to 14 days. In trial 1, use of the RH ≥97% threshold caused substantial differences between two RH sensors in recording number of hours exceeding the threshold. When both RH thresholds were compared for 2013–2015, on average, RH ≥90% resulted in a 53% reduction in variation of cumulative hours between two identical RH sensors placed adjacent to each other in an apple tree canopy. Although both the SBFS warning system and the calendar-based system resulted in equivalent control of SBFS, the warning system required fewer fungicide sprays than the calendar-based system, with an average of 3.8 sprays per season (min = 2; max = 5) vs. 6.4 sprays per season (min = 5; max = 8), respectively. The two fungicide regimes provided equivalent SBFS control when used in conjunction with the warning system. A partial budget analysis showed that using the SBFS warning system with a threshold of RH ≥90% was cost effective for orchard sizes of >1 ha. The revised warning system has potential to become a valuable decision support tool for Midwest apple growers because it reduces fungicide costs while protecting apples as effectively as a calendar-based spray schedule. The next step toward implementation of the SBFS warning system in the North Central U.S. should be multiyear field testing in commercial orchards throughout the region.

The El Niño Southern Oscillation (ENSO) is an oceanic-atmospheric phenomenon influencing worldwide weather and climate. Its occurrence is determined by the sea surface temperature (SST) anomaly of the 3.4 Niño region in the Pacific Ocean (5°N–5°S, 120°–170°W). El Niño (EN), Neutral (NT), and La Niña (LN) are the three possible phases of ENSO, respectively, for warm, normal, and cold SST anomaly. As in other regions around the world, weather in Brazil is influenced by ENSO phases. The country is the major coffee producer in the world, and production is strongly influenced by weather conditions, which affect plant yield, harvest quality, and interactions with pests and diseases. Coffee leaf rust (CLR), caused by the fungus Hemileia vastatrix, is a major cause of coffee yield and quality losses in Brazil, and requires fungicide spray applications every season. Because CLR is highly influenced by weather conditions, it is possible to use weather variables to simulate its progress during the cropping cycle. Therefore, the aims of this study were to estimate CLR infection rate based on a validated empirical model, which has daily minimum air temperature and relative humidity as inputs, and to assess the extent of ENSO influence on the annual risk of this disease at 45 sites in Brazil. Cumulative infection rates (CIR) were estimated daily from October to June of each growing season and location, based on the prevailing ENSO phase. Differences between the extreme phases (EN-LN) were assessed by the Two-One-Sided-Tests (TOST) method. Analysis of data from eight sites, located mainly in Paraná State, provided evidence of CIR differences between EN and LN phases (G1). Evidence of no difference of CIR between EN and LN was found in 18 sites (G2), whereas 19 sites showed no evidence of differences (G3) due to relatively large variation of CIR within the same ENSO phase. The G1 sites are located mostly in Southern Brazil, where ENSO exerts a well-defined influence on rainfall regime. In contrast, the G2 sites are mainly in Minas Gerais State, which is characterized as a transition region for ENSO influence on rainfall. The G3 sites are located between the northern region of Minas Gerais State and southern region of Bahia State, which is characterized by a subhumid climate that is usually very dry during winter, and where rainfall can vary up to 300% from one year to another, influencing relative humidity and resulting in a high CIR variability. Therefore, ENSO had a well-defined influence on CIR only in Paraná State, a region with minor importance for coffee production in Brazil. No ENSO influence was found in more northerly zones where the majority of Brazilian coffee is produced. This is the first evidence of ENSO-linked regional impact on the risk of coffee rust.

Batzer, J. C., Gleason, M. L., Weldon, B., Dixon, P. M., and Nutter, F. W., Jr. 2002. Evaluation of postharvest removal of sooty blotch and flyspeck on apples using sodium hypochlorite, hydrogen peroxide with peroxyacetic acid, and soap. Plant Dis. 86:1325-1332. Postharvest dips of apples (Malus × domestica) in commercial disinfestants were used to remove signs of the flyspeck (FS) pathogen, Schizothyrium pomi, and the sooty blotch (SB) complex (Peltaster fructicola, Leptodontium elatius, and Geastrumia polystigmatis). Apples were dipped for 7 or 15 min in buffered sodium hypochlorite (Agclor 310 plus Decco 312 Buffer) at 200, 400, 500, 600, or 800 ppm chlorine, a mixture of hydrogen peroxide and peroxyacetic acid (Tsunami 100) at 60 ppm/80 ppm, 120 ppm/160 ppm, or 360 ppm/480 ppm, respectively, or soap (Kleen 440), then brushed and rinsed for 30 s on a commercial grading line. Disease severity was assessed as percent diseased area using a quantitative rating system, and by counting the number of colonies of three mycelial types of SB and FS. Percent diseased area on apples was converted to USDA apple grade ratings and retail values. Both assessment methods provided similar results, but the percent-diseased-area method was less labor intensive. A 7-min dip in 800 ppm chlorine resulted in a mean increase from 25 and 55% to 100% Extra Fancy grade for ‘Jonathan’ and ‘Golden Delicious’ apples, respectively, and increased market value by 31 and 14%, respectively. The 7-min, 200-ppm chlorine dip resulted in an increase from 28 and 45% to 92.5 and 96.5% Extra Fancy after treatment for ‘Jonathan’ and ‘Golden Delicious’, respectively. Blemishes were removed more effectively from ‘Jonathan’ and ‘McIntosh’ apples than from ‘Golden Delicious’. Mycelial types of the sooty blotch and flyspeck fungi were removed differentially by the disinfestant dip treatments.

Pethybridge, S. J., Esker, P., Dixon, P., Hay, F., Groom, T., Wilson, C., and Nutter, F. W., Jr. 2007. Quantifying loss caused by ray blight disease in Tasmanian pyrethrum fields. Plant Dis. 91:1116-1121. The efficacy of newly implemented fungicide recommendations on reducing the intensity of ray blight disease caused by Phoma ligulicola to achieve site-specific attainable yield potentials in Tasmanian pyrethrum fields was quantified over two seasons in 46 and 51 fields during the 2003 and 2004 growing seasons, respectively. Disease intensity and yield in two plots (10 × 24 m), one following the commercial fungicide protocol recommendations and the second receiving no fungicide, were assessed in each pyrethrum field. The commercial fungicide protocol consisted of one application of azoxystrobin at 150 g a.i./ha, followed by two applications of a tank mixture of difenoconazole at 125 g a.i./ha and chlorothalonil at 1,008 liters a.i./ha at 14- to 21-day intervals. This program resulted in significant decreases in defoliation severity and the incidence of stems and flowers with ray blight, and increases in the height of stems and number of flowers produced per stem in October and November. In plots receiving the commercial fungicide protocol, the dry weight of flowers was increased by 76 and 68% in 2003 and 2004, respectively. Moreover, pyrethrin yield increased by 81 and 78% when the commercial fungicide protocol was used compared with the nontreated plots. Tobit regression was used to examine the relationships and thresholds among disease intensity measures (defoliation severity, stem severity, and incidence of flowers with ray blight) assessed just prior to harvest. This regression utilized a leftcensored regression model to define subminimal thresholds, as none of the disease intensity measures could be less than 0. Defoliation severity had a threshold of 35.3% before stem severity linearly increased and a threshold of 38.2% before the incidence of flowers with ray blight linearly increased. Finally, the threshold for stem severity was 13.7% before the incidence of flowers with ray blight linearly increased. These thresholds can be used to assist growers in making disease management decisions with the objective of minimizing loss of flowers by maintaining defoliation severity below the critical point at which the incidence of flowers with ray blight begins to linearly increase.

Xu, Z., Gleason, M. L., Mueller, D. S., Esker, P. D., Bradley, C. A., Buck, J. W., Benson, D. M., Dixon, P. M., and Monteiro, J. E. B. A. 2008. Overwintering of Sclerotium rolfsii and S. rolfsii var. delphinii in different latitudes of the United States. Plant Dis. 92:719-724. Previously known only from the southern United States, hosta petiole rot recently appeared in the northern United States. Sclerotium rolfsii var. delphinii is believed to be the predominant petiole rot pathogen in the northern United States, whereas S. rolfsii is most prevalent in the southern United States. In order to test the hypothesis that different tolerance to climate extremes affects the geographic distribution of these fungi, the survival of S. rolfsii and S. rolfsii var. delphinii in the northern and southeastern United States was investigated. At each of four locations, nylon screen bags containing sclerotia were placed on the surface of bare soil and at 20-cm depth. Sclerotia were recovered six times from November 2005 to July 2006 in North Dakota and Iowa, and from December 2005 to August 2006 in North Carolina and Georgia. Survival was estimated by quantifying percentage of sclerotium survival on carrot agar. Sclerotia of S. rolfsii var. delphinii survived until at least late July in all four states. In contrast, no S. rolfsii sclerotia survived until June in North Dakota or Iowa, whereas 18.5% survived until August in North Carolina and 10.3% survived in Georgia. The results suggest that inability to tolerate low temperature extremes limits the northern range of S. rolfsii. Additional keywords: Hosta spp., ornamental crops

Although famous for photic courtship displays, fireflies (Coleoptera: Lampyridae) are also notable for emitting strong odors when molested. The identity of volatile emissions and their possible role, along with photic signals, as aposematic warnings of unpalatability have been little explored, especially in tropical species. Pursuant to the observation that the widespread Neotropical fireflies, Photuris trivittata and Bicellonycha amoena, emit pungent odors, glows, and flashes when handled, we investigated their cuticular and headspace chemistry. Gas chromatography–mass spectrometry analyses revealed that both fireflies have species-specific cuticular hydrocarbon profiles. Photuris trivittata headspace was dominated by 2-methoxy-3-(1-methylpropyl) pyrazine (hereafter, pyrazine), on the order of 1.59 ng/individual and a suite of sesquiterpenes, while B. amoena emitted 3-methoxy-2-butenoic acid methyl ester and a few ketones. This is the first report of such compounds in fireflies. We investigated the role of pyrazine in P. trivittata's interactions with potential predators: sympatric ants, toads, and bats. Solvent-washed P. trivittata painted with pyrazine incurred lower ant predation than did their solvent-washed counterparts. Pyrazine significantly repelled ants at baits in concentrations as low as 9.8 × 10−4 ng/μl. The toad, Rhinella marina, readily accepted intact fireflies, pyrazine-coated and uncoated mealworms. Both Myotis nigricans and Molossus molossus bats rejected fireflies, but accepted both pyrazine-coated and uncoated mealworms. While pyrazine repels ants, its role as an aposematic signal warning other potential predators of firefly distastefulness requires further investigation. Our results underscore the idea that multiple enemies exert conflicting selection on firefly defenses.

182 SNP markers were developed for Ayous (Triplochiton scleroxylon K. Schum) by incorporating information from two next generation sequencing approaches (RADseq Floragenex and AFLPseq IonTorrent PGM) into a single genotyping panel for MassARRAY® iPLEX™. This set of markers was successfully used to genotype 753 individuals from 43 populations in five Tropical West and Central African Countries. These loci have an expected heterozygosity range of 0.007–0.501 and F ST from 0 to 0.306.

Illegal logging is one of the largest illicit trades in the world, with high profits and generally low risks of detection and prosecution. Timber identification presents problems for law enforcement as traditionally used forensic methods such as wood anatomy and dendrochronology are often unable to confidently match wood evidence to the remains of illegally felled trees. Here we have developed and validated a set of genetic markers for individualisation in bigleaf maple (Acer macrophyllum), a high value timber species often felled illegally in the USA. Using 128 single nucleotide polymorphisms and three insertion/deletion markers developed through massively parallel sequencing, 394 individuals were genotyped on the MassARRAY® iPLEX™ platform (Agena Bio-science™, San Diego, USA) to produce a population reference database for the species. We demonstrate that the resulting DNA assay is reliable, species specific, effective at low DNA concentrations (

Class II Major Histocompatibility complex (MHC-II) is a polymorphic heterodimer that binds antigen-derived peptides and presents them on the surface of antigen presenting cells. This mechanism of antigen presentation leads to recognition by CD4 T-cells and T-cell activation, making it a critical element of adaptive immune response. For this reason, the structural determinants of MHC-II function have been of great interest for the past 30 years, resulting in a robust structural understanding of the extracellular regions of the complex. However, the membrane-localized regions have also been strongly implicated in protein-protein and protein-lipid interactions that facilitate Class II assembly, transport and function, and it is these regions that are the focus of this review. Here we describe studies that reveal the strong and selective interactions between the transmembrane domains of the MHC α, and invariant chains which, when altered, have broad reaching impacts on antigen presentation and Class II function. We also summarize work that clearly demonstrates the link between membrane lipid composition (particularly the presence of cholesterol) and MHC-II conformation, subsequent peptide binding, and downstream T-cell activation. We have integrated these studies into a comprehensive view of Class II transmembrane domain biology. [Abstract copyright: Copyright © 2018. Published by Elsevier Inc.]

The enzyme carboxypeptidase G2 (CPG2) is used in antibody‐directed enzyme prodrug therapy (ADEPT) to catalyse the formation of an active drug from an inert prodrug. Free CPG2 in the bloodstream must be inhibited before administration of the prodrug in order to avoid a systemic reaction in the patient. Although a few small‐molecule CPG2 inhibitors have been reported, none has been taken forward thus far. This lack of progress is due in part to a lack of structural understanding of the CPG2 active site as well as the absence of small molecules that can block the active site whilst targeting the complex for clearance. The work described here aimed to address both areas. We report the structural/functional impact of extensive point mutation across the putative CPG2 catalytic site and adjacent regions for the first time, revealing that residues outside the catalytic region (K208A, S210A and T357A) are crucial to enzyme activity. We also describe novel molecules that inhibit CPG2 whilst maintaining the accessibility of galactosylated moieties aimed at targeting the enzyme for clearance. This work acts as a platform for the future development of high‐affinity CPG2 inhibitors that occupy new chemical space and will advance the safe application of ADEPT in cancer treatment.\ud \ud

Muscular (i.e., quadriceps) weakness contributes to disease progression and precedes the appearance of patient-reported symptoms, such as pain and perceived physical dysfunction, in knee osteoarthritis (OA). It is unknown, however, if muscular-based and patient-reported outcomes differentially associate with systemic biomarkers reflective of the local mediators in knee OA. The purpose of this study was to identify if muscular-based and patient-reported outcomes differentially associate with circulating superoxide dismutase (SOD) and cytokines in knee OA. Subjects (n = 29) with pain, muscular weakness, and radiographic evidence (Kellgren-Lawrence grade ≥2) of knee OA in the involved (INV) leg were included in this study. Serum Cu/Zn and Mn SOD and cytokine concentrations were measured in fasting blood samples. Pain and physical dysfunction were subjectively assessed and muscle strength (i.e., peak isometric force and torque, and peak isokinetic-concentric knee-extension and -flexion torques) was determined unilaterally in the INV and non-involved (NI) legs. Peak isometric and peak isokinetic-concentric knee-flexion torques in the INV leg correlated with serum Cu/Zn SOD (both p 0.05). Peak isometric force and torque and peak isokinetic-concentric knee-extension and -flexion torques in the INV leg correlated with serum Mn SOD (all p 0.05). Pain and dysfunction inversely associated with serum IL-1β, IL-4, IL-5, IL-12, IL-13, and/or IFN-γ (p 0.05). Neither SOD associated with pain or dysfunction, and none of the cytokines associated with muscular-based outcomes. We conclude that common outcome measures used in the clinical evaluation of OA differentially associate with circulating SOD and cytokines.

The purpose of the presented protocols is to study the process of Au(III) binding to BSA, yielding conformation change-induced red fluorescence (λ(em) = 640 nm) of BSA-Au(III) complexes. The method adjusts the pH to show that the emergence of the red fluorescence is correlated with the pH-induced equilibrium transitions of the BSA conformations. Red fluorescent BSA-Au(III) complexes can only be formed with an adjustment of pH at or above 9.7, which corresponds to the "A-form" conformation of BSA. The protocol to adjust the BSA to Au molar ratio and to monitor the time-course of the process of Au(III) binding is described. The minimum number of Au(III) per BSA, to produce the red fluorescence, is less than seven. We describe the protocol in steps to illustrate the presence of multiple Au(III) binding sites in BSA. First, by adding copper (Cu(II)) or nickel (Ni(II)) cations followed by Au(III), this method reveals a binding site for Au(III) that is not the red fluorophore. Second, by modifying BSA by thiol capping agents, another nonfluorophore-forming Au(III) binding site is revealed. Third, changing the BSA conformation by cleaving and capping of the disulfide bonds, the possible Au(III) binding site(s) are illustrated. The protocol described, to control the BSA conformations and Au(III) binding, can be generally applied to study the interactions of other proteins and metal cations.

Alphaviruses are mosquito-transmitted RNA viruses which generally cause acute disease including mild febrile illness, rash, arthralgia, myalgia and more severely, encephalitis. In the mouse, peripheral infection with Semliki Forest virus (SFV) results in encephalitis. With non-virulent strains, infectious virus is detectable in the brain, by standard infectivity assays, for around ten days. As we have shown previously, in severe combined immunodeficient (SCID) mice, infectious virus is detectable for months in the brain. Here we show that in MHC-II-/- mice, with no functional CD4 T-cells, infectious virus is also detectable in the brain for long periods. In contrast, in the brains of CD8-/- mice, virus RNA persists but infectious virus is not detectable. In SCID mice infected with SFV, repeated intraperitoneal administration of anti-SFV immune serum rapidly reduced the titer of infectious virus in the brain to undetectable, however virus RNA persisted. Repeated intraperitoneal passive transfer of immune serum resulted in maintenance of brain virus RNA, with no detectable infectious virus, for several weeks. When passive antibody transfer was stopped, antibody levels declined and infectious virus was again detectable in the brain. In aged immunocompetent mice, previously infected with SFV, immunosuppression of antibody responses many months after initial infection also resulted in renewed ability to detect infectious virus in the brain. In summary, antiviral antibodies control and determine whether infectious virus is detectable in the brain but immune responses cannot clear this infection from the brain. Functional virus RNA capable of generating infectious virus persists and if antibody levels decline, infectious virus is again detectable.

Codium tenue, previously known reliably only from estuarine habitats in South Africa, is recorded from a similar habitat in the Walpole and Nornalup Inlet system, on the south coast of Western Australia. The Australian C. tenue has a repeatedly divaricately dichotomously branched thallus to 11.5 cm in height, with markedly compressed axes up to 1 cm in width at branch dichotomies, but distally attenuating to terete branch apices. Structurally, thalli have cortices with distinctive cuneate utricles up to 1310 μm long and 650 μm in diameter. Both the habit and structural morphology essentially agree with C. tenue as known in South Africa. Sequences generated from the Australian specimens are also wholly comparable with those of South African specimens newly generated in this study. While similarly disjunct South African/Western Australian distributions are known for other algae, that of C. tenue is particularly remarkable in that the species is apparently an estuarine specialist.

Numerous predictive models have been developed to determine the likelihood that non-native plants will escape from cultivation and potentially become invasive. Given the substantial biological and economic costs that can result from the introduction of a new invasive plant and the unending pressures of world trade and transport, the creation and implementation of effective predictive models are becoming increasingly important. One key question in the development of such models focuses on the geographic scope at which models can best be developed and applied. We have developed models to predict woody-plant naturalization in five local areas within the Upper Midwest (United States). Herein, we consider whether naturalization can be reasonably predicted from a single model for the entire region or whether local models are required for each specific area. We develop a random forest model to predict the probability of naturalization in the region and compare out-of-sample prediction errors between the regional and local models. The regional model makes better predictions of the probability of naturalization for those species observed to naturalize but worse predictions for those not currently observed to naturalize. This model development process has given us an opportunity (not previously addressed in the literature) to examine the strengths and weaknesses of local and regional approaches, with the ultimate intent of optimizing geographic scope.

Novel single nucleotide polymorphisms (SNPs) and insertion/deletions (INDELs) were identified for the Bigleaf maple (Acer macrophyllum Pursch) using a combined next generation sequencing approach on the Ion Torrent PGM system with genotyping on the MassARRAY® iPLEX™ platform. Five hundred and ninety-eight putative loci were identified through sequencing of DNA fragments following a double restriction enzyme digest method. Two hundred and four polymorphic loci (199 SNPs and five INDELs) were successfully amplified across 65 individuals from seven populations across the native range of the species. Twenty-nine loci showed evidence of deviation from Hardy–Weinberg Equilibrium, and 85 were significantly linked. Expected heterozygosity ranged from 0.015 to 0.472 and F ST from 0.011 to 0.359. These genetic resources will prove useful for future studies into the population genetics and phylogeography of this important and iconic timber species.

Codium fragile subsp. fragile (Suringar) Hariot is a large, dichotomously branched green alga that has invaded many temperate locations around the globe. Despite its wide distribution and pest status, there are few accounts of Codium fragile subsp. fragile that provide morphological descriptions, including illustrations, for comparison. In 2008, a seaweed sample suspected to be Codium fragile subsp. fragile was collected from Albany, Western Australia. As this taxon superficially resembles other local erect and dichotomously branching Codium species, a detailed morphological and molecular analysis of the suspect alga was conducted. Investigations confirmed the identity of this alga as the invasive subspecies Codium fragile subsp. fragile. This represents the first record of this species in Western Australia.

Topical 1,25-dihydroxyvitamin D (1,25D) and other vitamin D compounds have been shown to protect skin from damage by ultraviolet radiation (UVR) in a process that requires the vitamin D receptor. Yet, while mice which do not express the vitamin D receptor are more susceptible to photocarcinogenesis, mice unable to 1α-hydroxylate 25-hydroxyvitamin D to form 1,25D do not show increased susceptibility to UVR-induced skin tumors. A possible explanation is that an alternative pathway, which does not involve 1α-hydroxylation, may produce photoprotective compounds from vitamin D. The cholesterol side chain cleavage enzyme CYP11A1 is expressed in skin and produces 20-hydroxyvitamin D3 (20OHD) as a major product of vitamin D3. We examined whether topical 20OHD would affect UVR-induced DNA damage, inflammatory edema or immune suppression produced in Skh:hr1 mice. Photoprotection by 20OHD at 23 or 46pmol/cm(2) against cyclobutane pyrimidine dimers (DNA lesions) after UVR in mice was highly effective, up to 98±0.8%, (p

We report new findings on the red fluorescent (λem = 640 nm) bovine serum albumin (BSA)-gold (Au) compound initially described by Xie et al. (J. Am. Chem. Soc. 2009, 131, 888-889) as Au25 nanoclusters. The BSA-Au compounds were further reducible to yield nanoparticles, suggesting that these compounds were BSA-cationic Au complexes. We examined the correlations between BSA conformations (pH-induced as well as denatured) and the resulting fluorescence of BSA-Au complexes, to understand the possible cationic Au binding sites. The red fluorescence of the BSA-Au complex was associated with a particular isoform of BSA, the aged form (pH > 10) of the five pH-dependent BSA conformations, while the other conformations, expanded (pH < 2.7), fast (2.7 < pH < 4.3), normal (4.3 < pH < 8), and basic (8 < pH < 10) did not result in red fluorescence. There could be internal energy transfer mechanisms to produce red fluorescence, deduced from excitation-emission map measurements. The ensemble minimum number of Au(III) per BSA to yield red fluorescence was

Detection of somatic mutations in human leukocyte antigen (HLA) genes using whole-exome sequencing (WES) is hampered by the high polymorphism of the HLA loci, which prevents alignment of sequencing reads to the human reference genome. We describe a computational pipeline that enables accurate inference of germline alleles of class I HLA-A, B and C genes and subsequent detection of mutations in these genes using the inferred alleles as a reference. Analysis of WES data from 7,930 pairs of tumor and healthy tissue from the same patient revealed 298 nonsilent HLA mutations in tumors from 266 patients. These 298 mutations are enriched for likely functional mutations, including putative loss-of-function events. Recurrence of mutations suggested that these 'hotspot' sites were positively selected. Cancers with recurrent somatic HLA mutations were associated with upregulation of signatures of cytolytic activity characteristic of tumor infiltration by effector lymphocytes, supporting immune evasion by altered HLA function as a contributory mechanism in cancer.

Background Ulcerative colitis (UC) is a chronic inflammatory bowel disease. In UC, a wide range of criteria is used for disease remission, with few studies investigating the differences between disease remission and normal control groups. This paper compares known inflammatory and healing mediators in the mucosa of UC in clinical remission and normal controls, in order to better describe the remission state and the fundamental pathology Methods Mucosal biopsies from 72 study participants (44 UC and 24 normal controls) were included from the Advanced Study of inflammatory bowel disease (ASIB Study), Arctic University of Norway, Norway. Clinical remission was defined as Mayo clinical score ≤ 2, with endoscopic subscores of ≤ 1. Targeted gene transcription analyses were performed by quantitative polymerase chain reaction (qPCR) and using hydrolysis probes and SYBR-green. Results Among the mucosal transcripts examined, 10 genes were regulated in remission vs. normal controls, 8 upregulated pro-inflammatory transcripts (IL1B, IL33, TNF, TRAF1, CLDN2, STAT1, STAT3 and IL13Ra2) and 2 downregulated (pro-inflammatory TBX21 and anti-inflammatory TGFB1). Between the active and normal group, 5 additional genes were differently regulated CASP8, ADAM17, DFB1 and TFF3 were up-regulated while CHUK was down-regulated. Several master transcription factors for T-cell development (TBX21, GATA3, FOXP3, RORC and SPI1) and important inflammatory mediators (IL33, IL6, TGFB, IL10 and TLR4) were upregulated in patients with Mayo endoscopic score of 1 in comparison to 0. Conclusion The mucosa of UC in clinical and endoscopic remission differs from normal mucosa, suggesting a remaining dysregulation of inflammatory and wound healing mechanisms. In addition, there is a significant expression difference between mayo endoscopic score of 1 and 0. This difference indicates that t-cell differentiation is still ongoing in the Mayo endoscopic subscore (MES) of 1. Interestingly, TNF did not differ between MES 0 and 1 suggesting an alternative inflammatory pathway in low-grade UC inflammation.

G protein-coupled receptors (GPCRs) are the largest family of cell-surface receptors in mammals and facilitate a range of physiological responses triggered by a variety of ligands. GPCRs were thought to function as monomers, however it is now accepted that GPCR homo- and hetero-oligomers also exist and influence receptor properties. The Schizosaccharomyces pombe GPCR Mam2 is a pheromone-sensing receptor involved in mating and has previously been shown to form oligomers in vivo. The first transmembrane domain (TMD) of Mam2 contains a small-XXX-small motif, overrepresented in membrane proteins and well-known for promoting helix–helix interactions. An ortholog of Mam2 in Saccharomyces cerevisiae, Ste2, contains an analogous small-XXX-small motif which has been shown to contribute to receptor homo-oligomerization, localization and function. Here we have used experimental and computational techniques to characterize the role of the small-XXX-small motif in function and assembly of Mam2 for the first time. We find that disruption of the motif via mutagenesis leads to reduction of Mam2 TMD1 homo-oligomerization and pheromone-responsive cellular signaling of the full-length protein. It also impairs correct targeting to the plasma membrane. Mutation of the analogous motif in Ste2 yielded similar results, suggesting a conserved mechanism for assembly. Using co-expression of the two fungal receptors in conjunction with computational models, we demonstrate a functional change in G protein specificity and propose that this is brought about through hetero-dimeric interactions of Mam2 with Ste2 via the complementary small-XXX-small motifs. This highlights the potential of these motifs to affect a range of properties that can be investigated in other GPCRs.

Dendritic cell‐specific intercellular adhesion molecule 3‐grabbing non‐integrin‐related (DC‐SIGNR), also known as liver/lymph node‐specific intercellular adhesion molecule 3‐grabbing non‐integrin, CLEC4M, CD209L, and CD299, is a Ca2+‐dependent lectin that has been implicated in increasing the infection rates of several viruses, including HIV, but the physiological role of DC‐SIGNR in healthy cells is currently not known with certainty. A close homologue of DC‐SIGNR, dendritic‐cell specific intercellular adhesion molecule 3‐grabbing non‐integrin, has been shown to act as a recycling endocytic receptor, which binds pathogens at the cell's surface and then releases them in the low pH environment of endosomal compartments. However, it is currently under debate in the literature as to whether DC‐SIGNR plays a similar role. In this work, we used NMR to explore whether the DC‐SIGNR carbohydrate recognition domain (CRD) shows any pH dependence in its ability to bind carbohydrates and Ca2+. We found clear evidence of reduced or abolished CRD‐binding affinities for three different glycans at low pH (4.2) as compared to neutral pH (6.8). We also report the assignment of the DC‐SIGNR CRD in the apo form, and use these new results to characterize the degree of structural rearrangement upon binding (or release) of Ca2+. Finally, we report a differential effect of pH on the affinities of glycans containing mannose exclusively versus glycans containing GlcNAc moieties. Our results lead us to propose that the DC‐SIGNR CRD rapidly and reversibly releases glycan ligands and Ca2+ at reduced pH (behaviour that would be expected for an endocytic receptor), and that the binding of mannose‐containing oligosaccharides is more strongly affected by pH than the binding of GlcNAc‐containing oligosaccharides., DC‐SIGNR (CLEC4M) is a potential recycling receptor on hepatic endothelia. Solution NMR demonstrates alternate binding affinities for different glycans at low and neutral pH, and shows structural rearrangements upon binding & release of Ca2+ ions. DC‐SIGNR rapidly and reversibly releases glycan ligands and Ca2+ions at reduced pH, with oligomannose glycans showing greater pH sensitivity compared with GlcNAc‐containing glycans.

Summary Weed seeds in and on the soil are the primary cause of weed infestations in arable fields. Previous studies have documented reductions in weed seedbanks due to cropping system diversification through extended rotation sequences, but the impacts of different rotation systems on additions to and losses from weed seedbanks remain poorly understood. We conducted an experiment in Iowa, USA, to determine the fates of Setaria faberi and Abutilon theophrasti seeds in 2-, 3- and 4-year crop rotation systems when seed additions to the soil seedbank were restricted to a single pulse at the initiation of the study. Over the course of the experiment, seedlings were removed as they emerged and prevented from producing new seeds. After 41 months, seed population densities dropped >85% for S. faberi and >65% for A. theophrasti, but differences between rotation systems in the magnitude of seedbank reductions were not detected. Most of the reductions in seedbank densities took place from autumn through early spring in the first 5 months following seed deposition, before seedling emergence occurred, suggesting that seed predation and/or seed decay was important. For S. faberi, total cumulative seedling emergence and total seed mortality did not differ between rotation systems. In contrast, for A. theophrasti, seedling emergence was 71% lower and seed mortality was 83% greater in the 3- and 4-year rotation systems than in the 2-year system. Results of this study indicate that for certain weed species, such as A. theophrasti, crop rotation systems can strongly affect life-history processes associated with soil seedbanks.

The Sargassum subgenera Bactrophycus and Arthrophycus were considered to be geographically restricted to the northern and southern hemispheres, respectively. Phylogenetic analyses of 19 Sargassum subgenus Bactrophycus species and eight Sargassum subgenus Arthrophycus species, based on a concatenated dataset of the loci ITS-2, cox3 and the rbcL-S spacer, showed that they formed a single clade, with Arthrophycus species nested within Bactrophycus section Halochloa. We merged the two subgenera as subgenus Bactrophycus and transferred “Arthrophycus” species to Sargassum section Halochloa. The genus now includes only the two subgenera, Sargassum and Bactrophycus, and both were found at temperate and subtropical latitudes; only subgenus Sargassum occurred at low latitudes near the equator, whereas subgenus Bactrophycus had an antitropical, disjunct distribution.

The sap-sucking insects (order Hemiptera), including aphids, planthoppers, whiteflies and stink bugs, present one of the greatest challenges for pest management in global agriculture. Insect neurotoxins offer an alternative to chemical insecticides for controlling these pests, but require delivery into the insect hemocoel. Here we use the coat protein of a luteovirus, an aphid-vectored plant virus, to deliver a spider-derived, insect-specific toxin that acts within the hemocoel. The luteovirid coat protein is sufficient for delivery of fused proteins into the hemocoel of pea aphids, Acyrthosiphon pisum, without virion assembly. We show that when four aphid pest species-A. pisum, Rhopalosiphum padi, Aphis glycines and Myzus persicae-feed on a recombinant coat protein-toxin fusion, either in an experimental membrane sachet or in transgenic Arabidopsis plants, they experience significant mortality. Aphids fed on these fusion proteins showed signs of neurotoxin-induced paralysis. Luteovirid coat protein-insect neurotoxin fusions represent a promising strategy for transgenic control of aphids and potentially other hemipteran pests.

Terrestrial habitats surrounding isolated wetlands are a critical resource for many pond-breeding amphibian species, yet few studies have examined the terrestrial distribution of post-metamorphic juveniles and adults. We used an encircling drift fence at a breeding pond in conjunction with partial fences at 90, 172, and 332 m from the wetland to estimate the terrestrial distribution of adult marbled salamanders (Ambystoma opacum; four breeding seasons) and mole salamanders (A. talpoideum; two seasons), as well as the dispersion of newly metamorphosed A. opacum (one summer). For newly metamorphosed A. opacum, 79% emigrated ,90 m from the wetland, and 8% moved beyond 172 m; movement distance was unrelated to body size. Distribution of adult A. opacum varied among years, with an average of 28% (range 23-31%) occurring beyond 172 m in all years. Averaged across two years, 51% of adult A. talpoideum occurred beyond 172 m. Lognormal models provided a good fit to both the juvenile and adult ambystomatid distributions, and parameters differed between age classes, sexes, species, and years within species. For adult A. opacum a buffer radius of 300 m or 340 m, depending on the year, is estimated to include 95% of adults; for A. talpoideum the estimate is 464 m or 501 m. A reanalysis of distribution data for seven ambystomatid species shows that a previous estimate of a 164-m radius to protect 95% of a population underestimates the needed buffer radius by 185 m. Because our study wetland requires a nearly 500 m wide radius to protect 95% of its ambystomatid adults, preservation of similar communities may require much more surrounding terrestrial habitat than previously thought.