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This dataset contains all data on which the following publication below is based. Paper Citation: Risch Anita C., Zimmermann, Stefan, Schütz, Martin, Borer, Elizabeth T., Broadbent, Arthur A.D., Caldeira, Maria C., Davies, Kendi F., Eisenhauer, Nico, Eskelinen, Anu, Fay, Philip A., Hagedorn, Frank, Knops, Johannes M.H., Lembrechts, Jonas, J., MacDougall, Andrew S., McCulley, Rebecca L., Melbourne, Brett A., Moore, Joslin L., Power, Sally A., Seabloom, Eric W., Silveira, Maria L., Virtanen, Risto, Yahdjian, Laura, Ochoa-Hueso, Raul (accepted). Drivers of the microbial metabolic quotient across global grasslands. Global Ecology and Biogeography Please cite this paper together with the citation for the datafile. The microbial metabolic quotient (MMQ; mg CO2-C mg MBC-1 h-1), defined as the amount of microbial CO2 respired (MR; mg CO2-C kg soil-1 h-1) per unit of microbial biomass C (MBC; mg C kg soil-1), is a key parameter for understanding the microbial regulation of the carbon (C) cycle, including soil C sequestration. Here, we experimentally tested hypotheses about the individual and interactive effects of multiple nutrient addition (NPK+micronutrients) and herbivore exclusion on MR, MBC, and MMQ across 23 sites (5 continents). Our sites encompassed a wide range of edaphoclimatic conditions, thus we assessed which edaphoclimatic variables affected MMQ the most and how they interacted with our treatments. Soils were collected in plots with established experimental treatments. MR was assessed in a five-week laboratory incubation without glucose addition, MBC via substrate-induced respiration. MMQ was calculated as MR/MBC and corrected for soil temperatures (MMQsoil). Using LMMs and SEMs, we analysed how edaphoclimatic characteristics and treatments interactively affected MMQsoil. MMQsoil was higher in locations with higher mean annual temperature, lower water holding capacity, and soil organic C concentration, but did not respond to our treatments across sites as neither MR nor MBC changed. We attributed this relative homeostasis to our treatments to the modulating influence of edaphoclimatic variables. For example, herbivore exclusion, regardless of fertilization, led to greater MMQsoil only at sites with lower soil organic C (-1.7%). Our results pinpoint the main variables related to MMQsoil across grasslands and emphasize the importance of the local edaphoclimatic conditions in controlling the response of the C cycle to anthropogenic stressors. By testing hypotheses about MMQsoil across global edaphoclimatic gradients, this work also helps to align the conflicting results of prior studies.

Brain edema; Intracranial hemorrhage; Serum biomarker Edema cerebral; Hemorragia intracraneal; Biomarcador sérico Edema cerebral; Hemorràgia intracranial; Biomarcador sèric Background: Early identification of patients developing symptomatic intracranial hemorrhage and symptomatic brain edema after acute ischemic stroke is essential for clinical decision-making. Astroglial protein S-100B is a marker of blood-brain barrier disruption, which plays an important role in the formation of intracranial hemorrhage and brain edema. In this study, we assessed the prognostic value of serum S-100B for the development of these complications. Methods: Serum S-100B levels were measured within 24 h from symptom onset in 1749 consecutive acute ischemic stroke patients from the prospective, observational, multicenter BIOSIGNAL cohort study (mean age 72.0 years, 58.3% male). To determine symptomatic intracranial hemorrhage or symptomatic brain edema, follow-up neuroimaging was performed in all patients receiving reperfusion therapy or experiencing clinical worsening with an NIHSS increase of ⩾4. Results: Forty six patients (2.6%) developed symptomatic intracranial hemorrhage and 90 patients (5.2%) developed symptomatic brain edema. After adjustment for established risk factors, log10S-100B levels remained independently associated with both symptomatic intracranial hemorrhage (OR 3.41, 95% CI 1.7–6.9, p = 0.001) and symptomatic brain edema (OR 4.08, 95% CI 2.3–7.1, p

The NF-κB transcription factor c-Rel plays a crucial role in promoting and regulating immune responses and inflammation. However, the function of c-Rel in modulating the mucosal immune system is poorly understood. T follicular helper (Tfh) cells and IgA production in gut-associated lymphoid tissues (GALT) such as Peyer's patches (PPs) are important for maintaining the intestinal homeostasis. Here, c-Rel was identified as an essential factor regulating intestinal IgA generation and function of Tfh cells. Genetic deletion of c-Rel resulted in the aberrant formation of germinal centers (GCs) in PPs, significantly reduced IgA generation and defective Tfh cell differentiation. Supporting these findings, the Ag-specific IgA response to Citrobacter rodentium was strongly impaired in c-Rel-deficient mice. Interestingly, an excessive expansion of segmented filamentous bacteria (SFB) was observed in the small intestine of animals lacking c-Rel. Yet, the production of IL-17A, IgA, and IL-21, which are induced by SFB, was impaired due to the lack of transcriptional control by c-Rel. Collectively, the transcriptional activity of c-Rel regulates Tfh cell function and IgA production in the gut, thus preserving the intestinal homeostasis.

Additional co-authors: Ivan Ropovik, Balazs Aczel, Lena F. Aeschbach, Luca Andrighetto, Jack D. Arnal, Holly Arrow, Peter Babincak, Bence E. Bakos, Gabriel Banik, Ernest Baskin, Radomir Belopavlovic, Michael H. Bernstein, Michal Bialek, Nicholas G. Bloxsom, Bojana Bodroža, Diane B. V. Bonfiglio, Leanne Boucher, Florian Bruhlmann, Claudia C. Brumbaugh, Erica Casini, Yiling Chen, Carlo Chiorri, William J. Chopik, Oliver Christ, Antonia M. Ciunci, Heather M. Claypool, Sean Coary, Marija V. Cˇolic, W. Matthew Collins, Paul G. Curran, Chris R. Day, Anna Dreber, John E. Edlund, Filipe Falcao, Anna Fedor, Lily Feinberg, Ian R. Ferguson, Maire Ford, Michael C. Frank, Emily Fryberger, Alexander Garinther, Katarzyna Gawryluk, Kayla Ashbaugh, Mauro Giacomantonio, Steffen R. Giessner, Jon E. Grahe, Rosanna E. Guadagno, Ewa Halasa, Rias A. Hilliard, Joachim Huffmeier, Sean Hughes, Katarzyna Idzikowska, Michael Inzlicht, Alan Jern, William Jimenez-Leal, Magnus Johannesson, Jennifer A. Joy-Gaba, Mathias Kauff, Danielle J. Kellier, Grecia Kessinger, Mallory C. Kidwell, Amanda M. Kimbrough, Josiah P. J. King, Vanessa S. Kolb, Sabina Kolodziej, Marton Kovacs, Karolina Krasuska, Sue Kraus, Lacy E. Krueger, Katarzyna Kuchno, Caio Ambrosio Lage, Eleanor V. Langford, Carmel A. Levitan, Tiago Jesse Souza de Lima, Hause Lin, Samuel Lins, Jia E. Loy, Dylan Manfredi, Łukasz Markiewicz, Madhavi Menon, Brett Mercier, Mitchell Metzger, Venus Meyet, Jeremy K. Miller, Andres Montealegre, Don A. Moore, Rafal Muda, Gideon Nave, Austin Lee Nichols, Sarah A. Novak, Christian Nunnally, Ana Orlic, Anna Palinkas, Angelo Panno, Kimberly P. Parks, Ivana Pedovic, Emilian Pekala, Matthew R. Penner, Sebastiaan Pessers, Boban Petrovic, Thomas Pfeiffer, Damian Pienkosz, Emanuele Preti, Danka Puric, Tiago Ramos, Jonathan Ravid, Timothy S. Razza, Katrin Rentzsch, Juliette Richetin, Sean C. Rife, Anna Dalla Rosa, Kaylis Hase Rudy, Janos Salamon, Blair Saunders, Przemyslaw Sawicki, Kathleen Schmidt, Kurt Schuepfer, Thomas Schultze, Stefan Schulz-Hardt, Astrid Schutz, Ani N. Shabazian, Rachel L. Shubella, Adam Siegel, Ruben Silva, Barbara Sioma, Lauren Skorb, Luana Elayne Cunha de Souza, Sara Steegen, L. A. R. Stein, R. Weylin Sternglanz, Darko Stojilovic, Daniel Storage, Gavin Brent Sullivan, Barnabas Szaszi, Peter Szecsi, Orsolya Szoke, Attila Szuts, Manuela Thomae, Natasha D. Tidwell, Carly Tocco, Ann-Kathrin Torka, Francis Tuerlinckx, Wolf Vanpaemel, Leigh Ann Vaughn, Michelangelo Vianello, Domenico Viganola, Maria Vlachou, Ryan J. Walker, Sophia C. Weissgerber, Aaron L. Wichman, Bradford J. Wiggins, Daniel Wolf, Michael J. Wood, David Zealley, Iris Žeželj, Mark Zrubka, and Brian A. Nosek

In Brazil, the country with the highest plant species richness in the world, biodiverse savannas and grasslands – i.e., grassy ecosystems, which occupy 27% of the country – have historically been neglected in conservation and scientific treatments. Reasons for this neglect include misconceptions about the characteristics and dynamics of these ecosystems, as well as inconsistent or regionally restricted terminology that impeded a more adequate communication about Brazil's savannas and grasslands, both within the country and internationally. Toward improved communication and recognition of Brazil's diversity of ecosystems, we present the key drivers that control the main types of grassy ecosystems across Brazil (including in regions of the country where forests dominate). In doing so, we synthesize the main features of each grassy ecosystem in terms of physiognomy and ecological dynamics (e.g., relationships with herbivores and fire). We propose a terminology both for major grassland regions and for regionally relevant vegetation physiognomies. We also discuss terms associated with human land management and restoration of grassy ecosystems. Finally, we suggest key research needs to advance our understanding of the ecology and conservation values of Brazil's grassy ecosystems. We expect that a common and shared terminology and understanding, as proposed here, will stimulate more integrative research that will be fundamental to developing improved conservation and restoration strategies.

This is the first study exclusively dedicated to the study of Hebeloma in Greenland. It is based on almost 400 collections, the great majority of which were collected by three of the co-authors over a period of 40 years and were lodged in the fungarium of the Natural History Museum in Copenhagen. The material was identified using molecular and morphological methods. In total, 28 species were recognized, 27 belonging to three sections, H. sects Hebeloma, Denudata and Velutipes. One species sampled was new to science and is here described as H. arcticum. For all species, a description, a distribution map within Greenland and macro and microphotographs are presented. A key is provided for the 28 species. The distribution of species within Greenland is discussed. The findings are placed in the context of studies of arctic and alpine Hebeloma from other parts of the world where comparable data exist. Notably, H. grandisporum, H. louiseae and H. islandicum, previously only known from Romania, Svalbard, Iceland or Norway, respectively, have been found in Greenland. The latter is also the only species encountered that does not belong to any of the above sections. Hebeloma excedens and H. colvinii – for the latter we here publish the first modern description – are to date only known from continental North America and now Greenland.

In 1994 Corner published five new species within the genus Psathyrella, all having been collected on the Malay Peninsula between 1929 and 1930. Three of these species belong to the genus Hebeloma and with their vinaceous colored lamellae and spore print, when fresh, they belong to H. sect. Porphyrospora. Of these three species, only one, P. flavidifolia, was validly published and thus we herewith recombine it as H. flavidifolium. The other two species, P. splendens and P. verrucispora, are synonyms of H. parvisporum and H. lactariolens, respectively. We also describe a new Malayan species, H. radicans, which also belongs to H. sect. Porphyrospora. These findings confirm the western Pacific Rim as a diversity hotspot for H. sect. Porphyrospora. The records described within this paper, represent the first recognition that the genus Hebeloma, and indeed that members of the ectomycorrhizal Hymenogastraceae, are present on the Malay Peninsula.

The COVID-19 pandemic created increased interest in monitoring patients at home to allow timely recognition of health deteriorations. Hospital care is particularly demanding in these patients because of the necessity for isolation to avoid further spread of the disease. Therefore, home care is a preferred treatment setting for these patients. This is, to our knowledge, the first report indicating the potential of an affordable, contactless, and unobtrusive ambient sensor system for the detection of signs of health deterioration in a patient with COVID-19 by a caregiver from a distance. Prospective data acquisition and correlation of the data with clinical events were obtained from an 81-year-old senior with COVID-19 before and, in particular, over a period of 10 days prior to hospitalization. Clinical signs included weakness, increased respiration rate, sleep disturbances, and confusion. The visualization of a combination of this information on a dedicated dashboard allowed the caregiver to recognize a serious health deterioration that required a lifesaving hospitalization. The potential of such ambient sensor systems to detect signs of serious health deterioration in patients with COVID-19 opens new opportunities for use in asymptomatic or oligosymptomatic patients who live alone and are sent back to their homes for isolation in quarantine after diagnosis.

ObjectiveThe severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic challenges national health systems and the global economy. Monitoring of infection rates and seroprevalence can guide public health measures to combat the pandemic. This depends on reliable tests on active and former infections. Here, we set out to develop and validate a specific and sensitive enzyme linked immunosorbent assay (ELISA) for detection of anti-SARS-CoV-2 antibody levels.MethodsIn our ELISA, we used SARS-CoV-2 receptor-binding domain (RBD) and a stabilized version of the spike (S) ectodomain as antigens. We assessed sera from patients infected with seasonal coronaviruses, SARS-CoV-2 and controls. We determined and monitored IgM-, IgA- and IgG-antibody responses towards these antigens. In addition, for a panel of 22 sera, virus neutralization and ELISA parameters were measured and correlated.ResultsThe RBD-based ELISA detected SARS-CoV-2-directed antibodies, did not cross-react with seasonal coronavirus antibodies and correlated with virus neutralization (R2 = 0.89). Seroconversion started at 5 days after symptom onset and led to robust antibody levels at 10 days after symptom onset. We demonstrate high specificity (99.3%;N = 1000) and sensitivity (92% for IgA, 96% for IgG and 98% for IgM; > 10 days after PCR-proven infection;N = 53) in serum.ConclusionsWith the described RBD-based ELISA protocol, we provide a reliable test for seroepidemiological surveys. Due to high specificity and strong correlation with virus neutralization, the RBD ELISA holds great potential to become a preferred tool to assess thresholds of protective immunity after infection and vaccination.

Hebeloma radicosum, known for its long ‘root’ and membranous ring, has regularly been recorded in Japan and been the subject of many studies revolving around ectomycorrhizal fungi exposed to high levels of nitrogen compounds and tripartite associations between fungi, host trees and mammals, including moles, wood mice and shrews. However, the species recorded in Japan is in fact not H. radicosum but a closely related species, also in Hebeloma section Myxocybe, described here as Hebeloma sagarae. This mushroom is macroscopically very similar to Hebeloma radicosum, also with a long ‘root’ and membranous ring, but distinct molecularly and in its distribution. We also examine other ‘rooting’ Hebeloma species from Japan: H. luchuense and H. radicosoides; both are shown to be members of H. sect. Scabrispora. A fourth ‘rooting’ Hebeloma from Japan is shown to be the same as or a close relative of Hebeloma danicum originally described from Europe.

The human ATPase p97, also known as valosin containing protein or Cdc48, is a highly abundant AAA+ engine that fuels diverse energy-consuming processes in the human cell. p97 represents a potential target for cancer therapy and its malfunction causes a degenerative disease. Here, we monitor the enzymatic activity of p97 in real time via an NMR-based approach that allows us to follow the steps that couple ATP turnover to mechanical work. Our data identify a transient reaction intermediate, the elusive ADP.P-i nucleotide state, which has been postulated for many ATPases but has so far escaped direct detection. In p97, this species is crucial for the regulation of adenosine triphosphate turnover in the first nucleotide-binding domain. We further demonstrate how the enzymatic cycle is detuned by disease-associated gain-of-function mutations. The high-resolution insight obtained into conformational transitions in both protein and nucleotide bridges the gap between static enzyme structures and the dynamics of substrate conversion. Our approach relies on the close integration of solution- and solid-state NMR methods and is generally applicable to shed light on the mechanochemical operating modes of large molecular engines.

Transporters of the solute carrier 6 (SLC6) family mediate the reuptake of neurotransmitters such as dopamine, norepinephrine, serotonin, GABA, and glycine. SLC6 family members are 12 transmembrane helix‐spanning proteins that operate using the transmembrane sodium gradient for transport. These transporters assume various quaternary arrangements ranging from monomers to complex stoichiometries with multiple subunits. Dopamine and serotonin transporter oligomerization has been implicated in trafficking of newly formed proteins from the endoplasmic reticulum to the plasma membrane with a pre‐fixed assembly. Once at the plasma membrane, oligomers are kept fixed in their quaternary assembly by interaction with phosphoinositides. While it remains unclear how oligomer formation precisely affects physiological transporter function, it has been shown that oligomerization supports the activity of release‐type psychostimulants. Most recently, single molecule microscopy experiments unveiled that the stoichiometry differs between individual members of the SLC6 family. The present overview summarizes our understanding of the influence of plasma membrane constituents on transporter oligomerization, describes the known interfaces between protomers and discusses open questions., We review the current knowledge on oligomerization of the solute carrier 6 (SLC6) family, which includes the transporters for the neurotransmitters dopamine, serotonin, and GABA. Early biochemical studies suggested dimer and tetramer configurations and identified several interaction motifs. Not all interfaces could be confirmed. Oligomers seem to become defined during transporter trafficking, are dependent on the plasma membrane PIP2 content, and affect substrate efflux. Simulations are starting to contribute to our understanding of transporter oligomerization, whereas recent single molecule experiments refined the oligomeric state by finding several non‐exchanging configurations for SERT, but only monomers and dimers for DAT.

Tuft cells are a rare population of chemosensory cells at the mucosal surface epithelia of hollow organs. Their name-giving morphological feature is an apical tuft of stiff microvilli. Accordingly, the actin-binding protein, villin, was identified as one of the first tuft cell markers in immunohistochemical analysis. Unfortunately, villin expression is not restricted to tuft cells, but is also prominent e.g. in enterocytes, which limits the use of this gene as a marker and as an experimental tool to genetically target tuft cells. Here, we report that the villin-related protein, advillin, is a specific tuft cell marker in the gastro-intestinal and biliary tract epithelia. In situ hybridization and immunohistochemistry revealed that advillin expression, unlike villin, was restricted to solitary cholinergic tuft cells in the mucosal linings of the small and large intestine, and in the gall bladder. In the glandular stomach, villin and advillin mRNA were present in all epithelial cells, while detectable protein levels were confined to solitary tuft cells. Advillin expression was no longer detectable in the mucosa of the intestinal and biliary tract from Pou2f3 deficient mice that lack tuft cells. Finally, crossing Avil-Cre transgenic mice with a double-fluorescent reporter mouse line resulted in specific targeting of gastro-intestinal and biliary tuft cells. Our analysis introduces advillin as a selective marker and tool in histological and functional analysis of the alimentary tract tuft cell system. Electronic supplementary material The online version of this article (doi:10.1007/s10735-020-09893-6) contains supplementary material, which is available to authorized users.

In the version of this Article originally published, there were unit conversion errors in the dataset when calculating specific leaf area (SLA) values at 10 of the 27 sites; errors were made when converting SLA from cm 2 g –1 to mm 2 g –1 and from mm 2 mg –1 to mm 2 g –1. This resulted in two incorrect data points: Phleum pratense in plot 27 at the Frue.ch site (SLA >300,000 mm 2 g –1) and Poa secunda in plot 31 at the shps.us site (SLA >60 mm 2 g –1). These two values were changed to ‘NA’ in the dataset, and therefore some reported estimates were changed, resulting in changes to the data points in Fig. 2a, the top bar of Fig. 3 and several values in the text.

Species of the genus Trichoderma are filamentous fungi commonly used in research, industry and agriculture. Trichoderma reesei strains are prominent producers of cellulolytic and hemicellulolytic enzymes as well as being expression hosts; several other species such as T. atroviride might be exploited as biocontrol agents. A careful preparation of Trichoderma inocula, which consists mainly of conidia (asexual spores), is of immense importance. Conidia concentration is still mostly determined with the help of a hemocytometer; however, as a more accurate and time-saving alternative, absorbance can be used to estimate fungal spore counts. We established a spectrophotometric method for fast and reliable preparation of Trichoderma inocula by evaluating the effect of size, shape and pigmentation of the conidia at different wavelengths.

Monocyte-derived and tissue-resident macrophages are ontogenetically distinct components of the innate immune system. Assessment of their respective functions in pathology is complicated by changes to the macrophage phenotype during inflammation. Here, we find that Cxcr4-CreER enables permanent genetic labeling of hematopoietic stem cells (HSCs) and distinguishes HSC-derived monocytes from microglia and other tissue-resident macrophages. By combining Cxcr4-CreER-mediated lineage tracing with Cxcr4 inhibition or conditional Cxcr4 ablation in photothrombotic stroke, we find that Cxcr4 promotes initial monocyte infiltration and subsequent territorial restriction of monocyte-derived macrophages to infarct tissue. After transient focal ischemia, Cxcr4-deficiency reduces monocyte infiltration and blunts the expression of pattern recognition and defense response genes in monocyte-derived macrophages. This is associated with an altered microglial response and deteriorated outcomes. Thus, Cxcr4 is essential for an innate immune-system-mediated defense response after cerebral ischemia. We further propose Cxcr4-CreER as a universal tool to study functions of HSC-derived cells.

Numerous allelopathic plant secondary metabolites impact plant–microorganism interactions by injuring plant-associated beneficial bacteria and fungi. Fungi belonging to the genus Trichoderma positively influence crops, including benzoxazinone-containing maize. However, benzoxazinones and their downstream metabolites such as benzoxazolinone and phenoxazinones are often fungitoxic. Specimen Trichoderma viride F-00612 was found to be insensitive to 100-µM phenoxazinone and 500-µM benzoxazolinone. Screening of 46 additional specimens of ascomycetes revealed insensitivity to phenoxazinones among fungi that cause disease in benzoxazinone-producing cereal crops, whereas many other ascomycetes were highly sensitive. In contrast, most of the screened fungi were insensitive to benzoxazolinone. T. viride F-00612 was associated with bacteria and, thus, existed as a consortium. By contrast, Enterobacter species and Acinetobacter calcoaceticus were prominent in the original specimen, and Bacillus species predominated after antibiotic application. Prolonged cultivation of T. viride F-00612 in liquid medium and on Czapek agar in the presence of T. viride F-00612 consortium may contribute to the ability to manage benzoxazinone downstream products and facilitate BOA-6-OH degradation via nitration.

Hebeloma parvisporum is described as new and placed within H. sect. Porphyrospora. This mushroom is sold as an edible in markets of Laos under the local name “wai khom.” Hebeloma sect. Porphyrospora is discussed and expanded to include the species formerly included in the genus Anamika and recently transferred to Hebeloma. Hebeloma sect. Porphyrospora currently comprises 16 species, 14 of which are known only from the western Pacific and Indian subcontinent. All species in this section share the character of having red-brown spores when fresh, atypical for other sections of Hebeloma, which causes the lamellae to be red-brown. However, this red-brown color fades when the material is dried. The close links, morphologically and molecularly, between H. parvisporum and other members of H. sect. Porphyrospora, particularly H. victoriense, are shown.

For patients suffering from neurodegenerative disorders, the behavior and activities of daily living are an indicator of a change in health status, and home-monitoring over a prolonged period of time by unobtrusive sensors is a promising technology to foster independent living and maintain quality of life. The aim of this pilot case study was the development of a multi-sensor system in an apartment to unobtrusively monitor patients at home during the day and night. The developed system is based on unobtrusive sensors using basic technologies and gold-standard medical devices measuring physiological (e.g., mobile electrocardiogram), movement (e.g., motion tracking system), and environmental parameters (e.g., temperature). The system was evaluated during one session by a healthy 32-year-old male, and results showed that the sensor system measured accurately during the participant’s stay. Furthermore, the participant did not report any negative experiences. Overall, the multi-sensor system has great potential to bridge the gap between laboratories and older adults’ homes and thus for a deep and novel understanding of human behavioral and neurological disorders. Finally, this new understanding could be utilized to develop new algorithms and sensor systems to address problems and increase the quality of life of our aging society and patients with neurological disorders.

1- Plant damage by invertebrate herbivores and pathogens influences the dynamics of grassland ecosystems, but anthropogenic changes in nitrogen and phosphorus availability can modify these relationships. 2- Using a globally distributed experiment, we describe leaf damage on 153 plant taxa from 27 grasslands worldwide, under ambient conditions and with experimentally elevated nitrogen and phosphorus. 3- Invertebrate damage significantly increased with nitrogen addition, especially in grasses and non-leguminous forbs. Pathogen damage increased with nitrogen in grasses and legumes but not forbs. Effects of phosphorus were generally weaker. Damage was higher in grasslands with more precipitation, but climatic conditions did not change effects of nutrients on leaf damage. On average, invertebrate damage was relatively higher on legumes and pathogen damage was relatively higher on grasses. Community-weighted mean damage reflected these functional group patterns, with no effects of N on community-weighted pathogen damage (due to opposing responses of grasses and forbs) but stronger effects of N on community-weighted invertebrate damage (due to consistent responses of grasses and forbs). 4- Synthesis. As human-induced inputs of nitrogen and phosphorus continue to increase, understanding their impacts on invertebrate and pathogen damage becomes increasingly important. Our results demonstrate that eutrophication frequently increases plant damage and that damage increases with precipitation across a wide array of grasslands. Invertebrate and pathogen damage in grasslands is likely to increase in the future, with potential consequences for plant, invertebrate and pathogen communities, as well as the transfer of energy and nutrients across trophic levels. EEA Santa Cruz Fil: Ebeling, Anne. University of Jena. Institute of Ecology and Evolution; Alemania Fil: Strauss, Alex T. University of Minnesota. Department of Ecology, Evolution, and Behavior; Estados Unidos Fil: Strauss, Alex T. University of Georgia. Odum School of Ecology; Estados Unidos Fil: Adler, Peter B. Utah State University. Department of Wildland Resources and the Ecology Center; Estados Unidos Fil: Arnillas, Carlos Alberto. University of Toronto —Scarborough. Department of Physical and Environmental Sciences; Canadá Fil: Barrio, Isabel C. Agricultural University of Iceland. Faculty of Environmental and Forest Sciences; Islandia Fil: Biederman, Lori A. Iowa State University. Department of Ecology, Evolution, and Organismal Biology; Estados Unidos Fil. Borer, Elizabeth T. University of Minnesota. Department of Ecology, Evolution, and Behavior; Estados Unidos Fil: Bugalho, Miguel N. University of Lisbon. Centre for Applied Ecology (CEABN-InBIO). School of Agriculture; Portugal. Fil: Caldeira, Maria C. University of Lisbon. Forest Research Centre. School of Agriculture; Portugal. Fil: Cadotte, Marc W. University of Toronto Scarborough. Department of Biological Sciences; Canadá Fil: Peri, Pablo Luis. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Santa Cruz; Argentina. Fil: Peri, Pablo Luis. Universidad Nacional de la Patagonia Austral; Argentina. Fil: Peri, Pablo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Fil: Blumenthal, Dana M. USDA-ARS, Rangeland Resources & Systems Research Unit; Estados Unidos

Background Digital sensing devices have become an increasingly important component of modern biomedical research, as they help provide objective insights into individuals’ everyday behavior in terms of changes in motor and nonmotor symptoms. However, there are significant barriers to the adoption of sensor-enhanced biomedical solutions in terms of both technical expertise and associated costs. The currently available solutions neither allow easy integration of custom sensing devices nor offer a practicable methodology in cases of limited resources. This has become particularly relevant, given the need for real-time sensor data that could help lower health care costs by reducing the frequency of clinical assessments performed by specialists and improve access to health assessments (eg, for people living in remote areas or older adults living at home). Objective The objective of this paper is to detail the end-to-end development of a novel sensor recording software system that supports the integration of heterogeneous sensor technologies, runs as an on-demand service on consumer-grade hardware to build sensor systems, and can be easily used to reliably record longitudinal sensor measurements in research settings. Methods The proposed software system is based on a server-client architecture, consisting of multiple self-contained microservices that communicated with each other (eg, the web server transfers data to a database instance) and were implemented as Docker containers. The design of the software is based on state-of-the-art open-source technologies (eg, Node.js or MongoDB), which fulfill nonfunctional requirements and reduce associated costs. A series of programs to facilitate the use of the software were documented. To demonstrate performance, the software was tested in 3 studies (2 gait studies and 1 behavioral study assessing activities of daily living) that ran between 2 and 225 days, with a total of 114 participants. We used descriptive statistics to evaluate longitudinal measurements for reliability, error rates, throughput rates, latency, and usability (with the System Usability Scale [SUS] and the Post-Study System Usability Questionnaire [PSSUQ]). Results Three qualitative features (event annotation program, sample delay analysis program, and monitoring dashboard) were elaborated and realized as integrated programs. Our quantitative findings demonstrate that the system operates reliably on consumer-grade hardware, even across multiple months (>420 days), providing high throughput (2000 requests per second) with a low latency and error rate ( Conclusions Overall, this sensor recording software could be leveraged to test sensor devices, as well as to develop and validate algorithms that are able to extract digital measures (eg, gait parameters or actigraphy). The proposed software could help significantly reduce barriers related to sensor-enhanced biomedical research and allow researchers to focus on the research questions at hand rather than on developing recording technologies.

Fertilisation experiments have demonstrated that nutrient availability is a key determinant of biomass production and carbon sequestration in grasslands. However, the influence of nutrients in explaining spatial variation in grassland biomass production has rarely been assessed. Using a global dataset comprising 72 sites on six continents, we investigated which of 16 soil factors that shape nutrient availability associate most strongly with variation in grassland aboveground biomass. Climate and N deposition were also considered. Based on theory-driven structural equation modelling, we found that soil micronutrients (particularly Zn and Fe) were important predictors of biomass and, together with soil physicochemical properties and C:N, they explained more unique variation (32%) than climate and N deposition (24%). However, the association between micronutrients and biomass was absent in grasslands limited by NP. These results highlight soil properties as key predictors of global grassland biomass production and point to serial co-limitation by NP and micronutrients.

Probiotics consumption has been associated with various health promoting benefits, including disease prevention and even treatment by modulating gut microbiota. Contrary to this, probiotics may also overstimulate the immune system, trigger systemic infections, harmful metabolic activities, and promote gene transfer. In children, the fragile immune system and impaired intestinal barrier may boost the occurrence of adverse effects following probiotics' consumption. To overcome these health challenges, the key focus has been shifted toward non-viable probiotics, also called paraprobiotics. Cell wall polysaccharides, peptidoglycans, surface proteins and teichoic acid present on cell's surface are involved in the interaction of paraprobiotics with the host, ultimately providing health benefits. Among other benefits, paraprobiotics possess the ability to regulate innate and adaptive immunity, exert anti-adhesion, anti-biofilm, anti-hypertensive, anti-inflammatory, antioxidant, anti-proliferative, and antagonistic effects against pathogens, while also enhance clinical impact and general safety when administered in children in comparison to probiotics. Clinical evidence have underlined the paraprobiotics impact in children and young infants against atopic dermatitis, respiratory and gastrointestinal infections, in addition to be useful for immunocompromised individuals. Therefore, this review focuses on probiotics-related issues in children's health and also discusses the Lactobacillus and Bifidobacterium spp. qualities for qualifying as paraprobiotics and their role in promoting the children's health.

Prostate cancer (PCa) in dogs is a highly malignant disease akin to its human counterpart. In contrast to the situation in humans, multi-gene approaches facilitating risk stratification of canine PCa are barely established. The aims of this study were the characterization of the transcriptional landscape of canine PCa and the identification of diagnostic, prognostic and/or therapeutic biomarkers through a multi-step screening approach. RNA-Sequencing of ten malignant tissues and fine-needle aspirations (FNA), and 14 nonmalignant tissues and FNAs was performed to find differentially expressed genes (DEGs) and deregulated pathways. The 4098 observed DEGs were involved in 49 pathways. These 49 pathways could be grouped into five superpathways summarizing the hallmarks of canine PCa: (i) inflammatory response and cytokines, (ii) regulation of the immune system and cell death, (iii) cell surface and PI3K signaling, (iv) cell cycle, and (v) phagosome and autophagy. Among the highly deregulated, moderately to strongly expressed DEGs that were members of one or more superpathways, 169 DEGs were listed in relevant databases and/or the literature and included members of the PCa pathway, oncogenes, prostate-specific genes, and druggable genes. These genes are novel and promising candidate diagnostic, prognostic and/or therapeutic canine PCa biomarkers.

Modern sensor technology is increasingly used in older adults to not only provide additional safety but also to monitor health status, often by means of sensor derived digital measures or biomarkers. Social isolation is a known risk factor for late-life depression, and a potential component of social-isolation is the lack of home visits. Therefore, home visits may serve as a digital measure for social isolation and late-life depression. Late-life depression is a common mental and emotional disorder in the growing population of older adults. The disorder, if untreated, can significantly decrease quality of life and, amongst other effects, leads to increased mortality. Late-life depression often goes undiagnosed due to associated stigma and the incorrect assumption that it is a normal part of ageing. In this work, we propose a visit detection system that generalizes well to previously unseen apartments - which may differ largely in layout, sensor placement, and size from apartments found in the semi-annotated training dataset. We find that by using a self-training-based domain adaptation strategy, a robust system to extract home visit information can be built (ROC AUC = 0.773). We further show that the resulting visit information correlates well with the common geriatric depression scale screening tool (rho = -0.87, p = 0.001), providing further support for the idea of utilizing the extracted information as a potential digital measure or even as a digital biomarker to monitor the risk of late-life depression.

Safety along the food and feed supply chain is an emerging topic and closely linked to the ability to analytical trace the geographical origin of food or feed. In this study, ultra-performance liquid chromatography coupled with electrospray ionization quadrupole-time-of-flight mass spectrometry was used to trace back the geographical origin of 151 grain maize (Zea mays L.) samples from seven countries using a high resolution non-targeted metabolomics approach. Multivariate data analysis and univariate statistics were used to identify promising marker features related to geographical origin. Classification using only 20 selected markers with the Random Forest algorithm led to 90.5% correctly classified samples with 100 times repeated 10-fold cross-validation. The selected markers were assigned to the class of triglycerides, diglycerides and phospholipids. The marker set was further evaluated for its ability to separate between one sample class and the rest of the dataset, yielding accuracies above 89%. This demonstrates the high potential of the non-polar metabolome to authenticate the geographic origin of grain maize samples. Furthermore, this suggests that focusing on only a few lipids with high potential for grain maize authentication could be a promising approach for later transfer of the method to routine analysis.

Summary Cxcl12-null embryos have dysplastic, misaligned, and hyperplastic semilunar valves (SLVs). In this study, we show that CXCL12 signaling via its receptor CXCR4 fulfills distinct roles at different stages of SLV development, acting initially as a guidance cue to pattern cellular distribution within the valve primordia during the endocardial-to-mesenchymal transition (endoMT) phase and later regulating mesenchymal cell proliferation during SLV remodeling. Transient, anteriorly localized puncta of internalized CXCR4 are observed in cells undergoing endoMT. In vitro, CXCR4+ cell orientation in response to CXCL12 requires phosphatidylinositol 3-kinase (PI3K) signaling and is inhibited by suppression of endocytosis. This dynamic intracellular localization of CXCR4 during SLV development is related to CXCL12 availability, potentially enabling activation of divergent downstream signaling pathways at key developmental stages. Importantly, Cxcr7-/- mutants display evidence of excessive CXCL12 signaling, indicating a likely role for atypical chemokine receptor CXCR7 in regulating ligand bioavailability and thus CXCR4 signaling output during SLV morphogenesis., Graphical abstract, Highlights • Semilunar valves are dysplastic, misaligned, and hyperplastic in Cxcl12-null mice • Cells undergoing endoMT express punctate CXCR4 and require CXCL12 for guidance • CXCL12 regulates proliferation of valve mesenchymal cells after E12.5 • CXCR7 is required as a sink for CXCL12 during valve development, Using in vivo and in vitro analysis, Ridge et al. show that the CXCL12/CXCR4/CXCR7 signaling axis fulfills distinct roles at different stages of mouse semilunar valve development. Regulation of ligand availability and CXCR4 activity by CXCR7 may be key to enabling alternative functional outcomes from chemokine signaling within a single tissue.

Understanding the course of the antibody response directed to individual epitopes of SARS-CoV-2 proteins is crucial for serological assays and establishment of vaccines. Twenty-one synthetic peptides were synthesized that have ten amino acids overlap and cover the complete membrane (M) protein. Plasma samples from 32 patients having acute disease and 30 patients from the convalescent phase were studied. Only peptide M01 (aa 1–20) and to a lesser extent peptide M21 (aa 201–222) showed specific reactivity as compared to historical control plasma samples. Peptide M01 was recognized by IgM- (71.9%) and IgG-specific antibodies (43.8%) during the acute phase as early as day 8 PIO. In a longitudinal analysis, a higher reactivity was observed for the IgM response directed to peptide M01 following day 20 PIO as compared to earlier time points of the acute phase. In the convalescent phase, antibody reactivity to the two M-specific peptides was significantly lower (

Fast, reliable, and comprehensive biodiversity monitoring data are needed for environmental decision making and management. Recent work on fish environmental DNA (eDNA) metabarcoding shows that aquatic diversity can be captured fast, reliably, and non-invasively at moderate costs. Because water in a catchment flows to the lowest point in the landscape, often a stream, it can often collect traces of terrestrial species via surface or subsurface runoff along its way or when specimens come into direct contact with water (e.g., for drinking purposes). Thus, fish eDNA metabarcoding data can provide information on fish but also on other vertebrate species that live in riparian habitats. This additional data may offer a much more comprehensive approach for assessing vertebrate diversity at no additional costs. Studies on how the sampling strategy affects species detection especially of stream-associated communities, however, are scarce. We therefore performed an analysis on the effects of biological replication on both fish as well as (semi-)terrestrial species detection. Along a 2 km stretch of the river Mulde (Germany), we collected 18 1-L water samples and analyzed the relation of detected species richness and quantity of biological replicates taken. We detected 58 vertebrate species, of which 25 were fish and lamprey, 18 mammals, and 15 birds, which account for 50%, 24%, and 7% of all native species to the German federal state of Saxony-Anhalt. However, while increasing the number of biological replicates resulted in only 25% more detected fish and lamprey species, mammal, and bird species richness increased disproportionately by 69% and 84%, respectively. Contrary, PCR replicates showed little stochasticity. We thus emphasize to increase the number of biological replicates when the aim is to improve general species detections. This holds especially true, when the focus is on rare aquatic taxa or on (semi-)terrestrial species, the so-called ‘bycatch’. As a clear advantage, this information can be obtained without any additional sampling or laboratory effort when the sampling strategy is chosen carefully. With the increased use of eDNA metabarcoding as part of national fish bioassessment and monitoring programs, the complimentary information provided on bycatch can be used for biodiversity monitoring and conservation on a much broader scale.

Exposure of Zea maysseedlings to 0.5µM benzoxazolinone-4/5/6/7-OH allelochemicals elicits root zone specific reactions by formation of dark rings and spots in the zone of lateral roots,high catalase activity at root hairsand no visible defense reaction at the root tip. We studied short term effects on membrane lipids and fatty acids in maize root tips in comparison to those of Abutilontheophrastiafter BOA-6-OH incubations. We found a decrease mainly of phosphatidylinositol in Abutilon and phosphatidylcholinein maize after 10-30 min of exposure. In the youngest tissue, α-linoleic acid (18:2), decreased considerably in both species but amounts recovered within one hour.Disturbances in membrane phospholipid contents were balanced in both species within 30-60 min. The findings that triacylglycerols(TAGs) were also affected, but levels of maize diacylglycerols (DAG) were almost unchanged, suggestedthat fatty acids for membrane lipid regeneration are released from TAGs while resulting DAGs are buildings blocks for phospholipid reconstitution,concomitant with BOA-6-OH glucosylation. Theexpression of superoxide dismutase (SOD2)and of ER-bound oleoyldesaturase (FAD2-2) genes were contemporaneously up regulated in contrast to the catalase gene CAT1,while CAT3was arguably integrated at a later stage of the detoxification process. Major catalase activity seems to be attributed to root colonizing microorganisms. Immuno-responses were not elicited in short-terms, since expression of NPR1, POX12were rather affected,PR4 after 6h with BOA-4/7-OH and PR1 after 24hwith BOA-5/6-OH.The rapid membrane recovery, reactive oxygen species and allelochemical detoxification may be characteristic forBOA-OH insensitive plants.

Wading birds play an important role in coastal and wetland ecosystems. As a result of anthropogenic disturbances, numbers of wading birds have declined over the past years, and therefore, the monitoring of bird species is crucial to preserve their habitats and counteract this. Due to known limitations of conventional monitoring, molecular approaches are increasingly becoming complementary methods. Thus, we evaluated the potential of an environmental DNA (eDNA) monitoring approach for wading birds. We developed three species-specific primer sets for wading birds targeting the COI region. After examining the primers in silico and on extracted bird DNA, the primers were used to successfully amplify eDNA from water samples.

Downregulation of ubiquitin (Ub) ligase activity prevents premature ubiquitination and is critical for cellular homeostasis. Nedd4 Ub ligases share a common domain architecture and yet are regulated in distinct ways through interactions of the catalytic HECT domain with the N-terminal C2 domain or the central WW domain region. Smurf1 and Smurf2 are two highly related Nedd4 ligases with ~70% overall sequence identity. Here, we show that the Smurf1 C2 domain interacts with the HECT domain and inhibits ligase activity in trans. However, in contrast to Smurf2, we find that full-length Smurf1 is a highly active Ub ligase, and we can attribute this striking difference in regulation to the lack of one WW domain (WW1) in Smurf1. Using NMR spectroscopy and biochemical assays, we identified the WW1 region as an additional inhibitory element in Smurf2 that cooperates with the C2 domain to enhance HECT domain binding and Smurf2 inhibition. Our work provides important insights into Smurf regulation and highlights that the activities of highly related proteins can be controlled in distinct ways.

Dimerization or the formation of higher-order oligomers is required for the activation of ErbB receptor tyrosine kinases. The heregulin (HRG) receptor, ErbB3, must heterodimerize with other members of the family, preferentially ErbB2, to form a functional signal transducing complex. Here, we applied single molecule imaging capable of detecting long-lived and mobile associations to measure their stoichiometry and mobility and analyzed data from experiments globally, taking the different lateral mobility of monomeric and dimeric molecular species into account. Although ErbB3 was largely monomeric in the absence of stimulation and ErbB2 co-expression, a small fraction was present as constitutive homodimers exhibiting a ∼40% lower mobility than monomers. HRG stimulation increased the homodimeric fraction of ErbB3 significantly and reduced the mobility of homodimers fourfold compared to constitutive homodimers. Expression of ErbB2 elevated the homodimeric fraction of ErbB3 even in unstimulated cells and induced a ∼2-fold reduction in the lateral mobility of ErbB3 homodimers. The mobility of ErbB2 was significantly lower than that of ErbB3, and HRG induced a less pronounced decrease in the diffusion coefficient of all ErbB2 molecules and ErbB3/ErbB2 heterodimers than in the mobility of ErbB3. The slower diffusion of ErbB2 compared to ErbB3 was abolished by depolymerizing actin filaments, whereas ErbB2 expression induced a substantial rearrangement of microfilaments, implying a bidirectional interaction between ErbB2 and actin. HRG stimulation of cells co-expressing ErbB3 and ErbB2 led to the formation of ErbB3 homodimers and ErbB3/ErbB2 heterodimers in a competitive fashion. Although pertuzumab, an antibody binding to the dimerization arm of ErbB2, completely abolished the formation of constitutive and HRG-induced ErbB3/ErbB2 heterodimers, it only slightly blocked ErbB3 homodimerization. The results imply that a dynamic equilibrium exists between constitutive and ligand-induced homo- and heterodimers capable of shaping transmembrane signaling.

INTRODUCTION: The WHO classification of pulmonary neuroendocrine tumors (PNETs) is also used to classify thymic NETs (TNETs) into typical and atypical carcinoid (TC and AC), large cell neuroendocrine carcinoma (LCNEC), and small cell carcinoma (SCC), but little is known about the usability of alternative classification systems. METHODS: One hundred seven TNET (22 TC, 51 AC, 28 LCNEC, and 6 SCC) from 103 patients were classified according to the WHO, the European Neuroendocrine Tumor Society, and a grading-related PNET classification. Low coverage whole-genome sequencing and immunohistochemical studies were performed in 63 cases. A copy number instability (CNI) score was applied to compare tumors. Eleven LCNEC were further analyzed using targeted next-generation sequencing. Morphologic classifications were tested against molecular features. RESULTS: Whole-genome sequencing data fell into three clusters: CNIlow, CNIint, and CNIhigh. CNIlow and CNIint comprised not only TC and AC, but also six LCNECs. CNIhigh contained all SCC and nine LCNEC, but also three AC. No morphologic classification was able to predict the CNI cluster. Cases where primary tumors and metastases were available showed progression from low-grade to higher-grade histologies. Analysis of LCNEC revealed a subgroup of intermediate NET G3 tumors that differed from LCNEC by carcinoid morphology, expression of chromogranin, and negativity for enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2). CONCLUSIONS: TNETs fall into three molecular subgroups that are not reflected by the current WHO classification. Given the large overlap between TC and AC on the one hand, and AC and LCNEC on the other, we propose a morphomolecular grading system, Thy-NET G1-G3, instead of histologic classification for patient stratification and prognostication. peerReviewed

Bacteria often express numerous virulence factors. These virulence factors make them successful pathogens, by e.g. mediating attachment to host cells and thereby facilitating persistence or invasion, or by contributing to the evasion of the host immune system to allow proliferation and spread within the host and in the environment. The site of first contact of Gram negative bacteria with the host is the bacterial outer membrane (OM). Consisting of an asymmetrical lipid bilayer with phospholipids forming the inner, and lipopolysaccharides forming the outer leaflet, the OM harbors numerous integral membrane proteins that are almost exclusively β-barrel proteins. One distinct family of OM β-barrel proteins strongly linked to bacterial virulence are the autotransporter (AT) proteins. During the last years huge progress has been made to better understand the mechanisms underlying the insertion of AT proteins into the OM and also AT function for interaction with the host. This review shortly summarizes our current knowledge about outer membrane protein (OMP) and more specifically AT biogenesis and function. We focused on the AT proteins that we haved studied in most detail: i.e. the Yersinia adhesin A (YadA) and invasin of Yersinia enterocolitica (Ye) as well as its homolog intimin (Int) expressed by enteropathogenic Escherichia coli. In addition, this review provides a short outlook about how we could possibly use this knowledge to fight infection.

ZusammenfassungDas physiologische intestinale Mikrobiom (Bakterien des Darmes) ist wesentlich für Funktion und Integrität des Darmes. Mikrobiom-Abweichungen können für die onkologische Therapie wichtige Hinweise liefern: Ein vermehrtes Auftreten des als invasiv bekannten Sulfatreduzierers Fusobacterium nucleatum (FN) im Mikrobiom zeigt eine hohe Korrelation mit dem Auftreten des kolorektalen Karzinoms. Hintergrund ist die Eigenschaft von FN, am Darmepithel ein entzündliches Mikroklima zu erzeugen, das den Darm schädigt und die Tumorprogression ebenso fördert wie die durch FN erzeugte Freisetzung des Transkriptionsfaktors NF-κB über die Expression von Onkogenen und weiteren proinflammatorischen Genprodukten. Darüber hinaus lässt FN die T-Zell-Dichte und -Aktivität sinken und hemmt so die Tumorabwehr. Weiterhin aktivieren proentzündliche Zytokine den Abbau der Aminosäure Tryptophan (TRP) zu Kynurenin. Damit wird einerseits TRP der Proteinbio- und der Serotoninsynthese entzogen. Andererseits sind erhöhte Kynurenin-Spiegel mit verringerten Überlebensraten assoziiert, da Kynurenin immunsuppressiv wirkt. Eine Therapie, die auf den Darm, seine Mikrobiota und das Eindämmen der Entzündung abzielt, kann die negativen Auswirkungen eingrenzen und die Regeneration und Remission des Patienten unterstützen.

Volatile organic compounds (VOCs) from alive Anoplophora glabripennis (ALB) imagoes, larvae and ovipositions were obtained by enrichment on adsorbents. They were analysed qualitatively by thermal desorption coupled with gas chromatography and a mass selective detector. All together 229 substances were identified: 34 from larvae, 19 from imago and 176 from oviposition. Ovipositions of two generations were analysed, both over an 8 week period with focus on aging and repeatability. (+)-α-Longipinene occurred in 100%, (+)-Cyclosativene and α-Cubebene in 99% of all oviposition measurements. 1R-α-Pinene occurred in 100% of all imago measurements. 2,4-Dimethyl-1-Heptene and (+)-Cyclosativene were detectable in all three ALB sample types.

The human dopamine transporter (hDAT) regulates the level of the neurotransmitter dopamine (DA) in the synaptic cleft and recycles DA for storage in the presynaptic vesicular pool. Many neurotransmitter transporters exist as oligomers, but the physiological role of oligomerization remains unclear; for example, it has been speculated to be a prerequisite for amphetamine-induced release and protein trafficking. Previous studies point to an oligomeric quaternary structure of hDAT; however, the exact stoichiometry and the fraction of co-existing oligomeric states are not known. Here, we used single-molecule brightness analysis to quantify the degree of oligomerization of heterologously expressed hDAT fused to monomeric GFP (mGFP–hDAT) in Chinese hamster ovary (CHO) cells. We observed that monomers and dimers of mGFP–hDAT co-exist and that higher-order molecular complexes of mGFP–hDAT are absent at the plasma membrane. The mGFP–hDAT dimers were stable over several minutes, and the fraction of dimers was independent of the mGFP–hDAT surface density. Furthermore, neither oxidation nor depletion of cholesterol had any effect on the fraction of dimers. Unlike for the human serotonin transporter (hSERT), in which direct binding of phosphatidylinositol 4,5-bisphosphate (PIP2) stabilized the oligomers, the stability of mGFP–hDAT dimers was PIP2 independent.

Only a few cobalamin-producing bacterial species are known which are suitable for food fermentations. The strain of Acetobacter pasteurianus DSM 3509 was found to have the capability to synthesize cobalamin. A survival test and a preliminary genetic study of the gene of uroporphyrinogen-III synthase indicated the ability to synthesize cobalamin. By a modified microbiological assay based on Lactobacillus delbrueckii ssp. lactis DSM 20355, 4.57 ng/mL of cyanocorrinoids and 0.75 ng/mL of noncorrinoid growth factors were detected. The product extracted and isolated by immunoaffinity chromatography in its cyanide form had the similar UV spectrum as standard cyanocobalamin and Coα-[α-(7-adenyl)]-(Coβ-cyano) cobamide also known as pseudovitamin B12 produced by Lactobacillus reuteri DSM 20016. The chromatographically separated product of A. pasteurianus was subjected to mass spectrometrical analysis. There, its fragmentation pattern turned out to be equivalent to that of cyanocobalamin also produced by Propionibacterium freudenreichii ssp. freudenreichii DSM 20271 and clearly differs from pseudovitamin B12. Due to the presence of this species in several food applications, there might be cobalamin residues in food fermented with these bacteria.

Numerous taxa ofHebelomahave been reported in association withSalix,Dryas, andBetulain arctic-alpine habitats. However, species are notoriously difficult to delineate because morphological features overlap, and previously there was little reliable molecular data available. Recent progress in ITS-sequencing within the genus, coupled with an extensive database of parametrically described collections, now allows comparisons between species and their distributions. Here we report 16 species ofHebelomafrom the Rocky Mountain alpine zone from some of the lowest latitudes (latitude 36°–45°N) and highest elevations (3000–4000 m) for arctic-alpine fungi in the northern hemisphere. Twelve of these species have been reported from arctic-alpine habitats in Europe and Greenland and are now molecularly confirmed from the Middle and Southern Rockies, greatly expanding their distribution. These are:Hebelomaalpinum,H.aurantioumbrinum,H.dunense,H.hiemale,H.marginatulum,H.mesophaeum,H.nigellum,H.oreophilum,H.subconcolor,H.spetsbergense,H.vaccinum, andH.velutipes. Hebelomahygrophilumis known from subalpine habitats in Europe, but was never recorded in arctic-alpine ecology. Three species recorded from the Rockies, but as yet not reported from Europe, areH.alpinicola,H.avellaneum, andH.excedens. The last two have never previously been reported from an arctic-alpine habitat. For all three of these species, the holotypes have been studied morphologically and molecularly, and have been incorporated into the analysis.

We used an information theoretic approach to assess the effects of an ecologically adjusted sediment management scheme on grayling (Thymallus thymallus L. 1758) populations. Additionally to reservoir operation, candidate models included a variety of parameters and processes that may influence grayling populations such as flow, temperature, density dependence, and bird predation. Population parameters analyzed included total densities, young of the year numbers, and larval densities. These analyses were supplemented by a characterization of sediments and sedimentation patterns in the reach. Investigations were carried out in six sites affected by flushing and in one control site. A total of thirteen flushing operations have been undertaken within the study period leading to considerable remobilization of fine sediments and gravel. Due to seasonal and hydrological restrictions, not every flood could be used for flushing. These limitations led to an interrupted management throughout the chain of reservoirs as well as to long time intervals between flushing events with possible effects on spawning habitat quality. None of the investigated population parameters was affected by flushing, and thus, the study generally supports the current reservoir management scheme. Our analyses revealed the magnitude and timing of high water events, temperature, and density‐dependent effects, that is, population densities the year before, as the most influential variables for grayling population dynamics in the investigated stretch. The siltation of reservoirs is a significant problem for reservoir storage, flood protection, river deltas, and coastal zones. Its management—which is inevitable to safeguard river deltas and secure flood protection—poses also the challenge to safeguard riverine ecosystems below reservoirs. Based on our experience, we propose a periodic flushing regime in concordance with the hydrograph thereby mimicking the timing, magnitude, frequency, and duration of natural SSC pulses and gravel transport. This flushing regime minimizes adverse downstream environmental impacts and maximizes benefits.

The utilization of fluorescent ligands to study the monoamine transporters (MATs) has increased our knowledge of their function and distribution in live cell systems. In this study, we extend SAR for nisoxetine and talopram as parent compounds, to identify high affinity rhodamine-labeled fluorescent probes for the norepinephrine transporter (NET). Nisoxetine-based fluorescent probe 6 demonstrated high binding affinity (K(i) = 43 nM) for NET and an overall selectivity compared to the other transporters for dopamine (DAT; K(i) = 1540 nM) and serotonin (SERT; K(i) = 785 nM) in competitive radioligand binding assays. Using confocal microscopy, compound 6 was shown to stain both NET and SERT, but not DAT, at low nanomolar concentrations, in transporter-expressing cells.