This study seeks to gain initial insight into what is talked about and whose voices are heard at Medical Library Association (MLA) annual meetings.Meeting abstracts were downloaded from the MLA website and converted to comma-separated values (CSV) format. Descriptive analysis in Python identified the number of presentations, disambiguated authors, author collaboration, institutional affiliation type, and geographic affiliation. Topics were generated using Mallet's Latent Dirichlet Allocation algorithm for topic modeling.There were 5,781 presentations at MLA annual meetings from 2001–2019. Author disambiguation resulted in approximately 5,680 unique authors. One thousand ninety-three records included a hospital-related keyword in the author field, and 4,517 records included an academic-related keyword. There were 438 presentations with at least 1 international author. The topic model identified 16 topics in the MLA abstract corpus: events, electronic resources, publications, evidence-based practice, collections, academic instruction, librarian roles and relationships, technical systems, special collections, general instruction, literature searching, surveys, research support, community outreach, patient education, and library services.Academic librarians presented more frequently than hospital librarians, though more research should be done to determine if this discrepancy was disproportionate to hospital librarians' representation in MLA. Geographic affiliation was concentrated in the United States and appeared to be related to population density. Health sciences librarians in the early twenty-first century are spending more time at MLA annual meetings talking about communities, relationships, and visible services, and less time talking about library collections and operations. Further research will be needed to boost the participation of underrepresented members. Professional conferences enable librarians and library staff to share projects, trends, resources, and ideas; network with their colleagues; and build relationships. Librarians consider conferences a valuable use of their time and an opportunity for “professional rejuvenation” . The MedJournal of the Medical Library Association (JMLA), previously known as the Bulletin of the Medical Library Association (BMLA). JMLA and other journals serve as platforms for professional discourse and preserve the scholarship of the profession. However, research suggests that librarians are more likely to present at a conference than publish a peer-reviewed article [MLA also has a peer-reviewed journal, the  article , 3. Libr article , and man article . The pubBMLA and JMLA articles to identify key topics from 1961–2010 [The Janet Doe lecture, presented by Mark E. Funk, AHIP, FMLA, at the 2012 MLA annual meeting, analyzed the content of Knowing what librarians talk about at MLA annual meetings is important because what we talk about is what we are doing, and what we are doing is creating the future work environments, services, and relationships that will shape and define libraries. Knowing whose voices are heard at annual meetings is important because MLA has identified diversity and inclusion as a top priority . FurtherThe purpose of this study was to use computational methods to describe the MLA annual meeting abstracts by analyzing the number of presentations, presentation dates, author information, and abstract content. Meeting abstracts were downloaded from the MLA website, which had portable document format (PDF) versions of meeting programs available for the years 2001 to 2018 . The PDFAfter conversion to text format, most of the meeting files contained a mix of line breaks, semicolons, and numbering to separate the presentations. Structured fields were created from this unstructured data through manually modifying meeting information, such as removing dates and locations, and by using regular expressions to extract presentation information and create consistent line breaks. Semicolons in the unstructured data were consistently used to separate each author's information, but the provided information varied between authors and was not consistently ordered within the semicolon separators. As an example, a record's author section might look like:Author name, job title, institution name, institution city, institution state;Author name, library name, institution name, institution city;Author name, job title, department, institution name, institution city and stateAfter data cleaning, the meeting programs contained the following fields: Title, Author (including affiliation), Abstract, Type (poster or paper), Year, and uniform resource locator (URL), links to the PDF version of the meeting program. The complete CSV file is available to browse and search online .Basic descriptive analysis was done using Python pandas . Author university, school, college, and department. For hospitals, the following keywords were used: hospital, health system, clinic, health center, and medical center. These searches were not mutually exclusive. Some abstracts had hospital librarians presenting alongside academic librarians, while other abstracts contained affiliation information such as “University Medical Center.”Institutional names were not normalized due to the highly variable representation of information in the author column. Inconsistent ordering and punctuation in the descriptions of job titles, institution names, library or department names, and geographic locations, as described above, prevented straightforward automation of a standardized institution field. Since it was beyond the scope of this study to manually extract and reconcile accurate institutional affiliations for each author with authority files, records were instead searched for common keywords related to institution type. For academic institutions, the Author field was searched for the following keywords: university and counted in the academic result set and would be matched on the phrase medical center and counted in the hospital result set. Identifying and categorizing other institution types was not attempted in this study due to a lack of standard descriptive keywords.For each abstract, the first occurrence of any of the academic-related keywords added the record to the academic authorship results, and the first occurrence of any of the hospital-related keywords added the record to the hospital authorship results. Records that contained keywords in the author field from both the academic- and hospital-related keyword lists were counted in both result sets, so an author from “University Medical Center” would be matched for the word Geographic affiliation was not consistently described in the abstracts. Author fields did not commonly indicate “United States,” “U.S.,” or “USA,” and more often they simply listed the US state of the institution. To determine geographic affiliation, a list of all the US states and territories, as well as state abbreviations, was compared to the Author field to extract the first occurrence of any state name. The resulting list counted the instances of each state name occurring at least once in each Author field. For example, a record containing three authors from Georgia and one author from Florida would be counted once for Georgia and once for Florida. A map was generated using Plotly's choropleth maps library  and can A similar approach was used for counting international contributions to MLA meetings. A list of all countries was compared to the Author field to extract the first occurrence of any country name in any Author field. These results were manually reviewed to ensure that personal names or American place names were not included. After double checking that no genuine records existed, the following countries were removed from the list: Georgia, Jamaica, Jordan, Lebanon, and Monaco. As with the institutional and US state search, these results were not mutually exclusive, so international authors who coauthored with each other had their presentations counted once for each represented country.The descriptive analysis code is available online in a Jupyter notebook .The abstracts were analyzed using topic modeling. Topic modeling is a method of computationally identifying topics in texts. It is an unsupervised approach, not requiring predetermined lists of categories, which is useful for initial exploration of textual data. For this analysis, the topic modeling algorithm chosen was Latent Dirichlet Allocation (LDA) . In simpAfter importation, data were cleaned and preprocessed. The data preprocessing was done with the Python libraries spacy , NLTK 2, and GenThe topics were visualized using pyLDAvis . This viJMLA from January 2001–July 2019, an average of 65 items per year.Presentations over time are shown in Fuzzy matching was used to disambiguate an original count of 6,732 author names, as described in the methods section, which resulted in approximately 5,680 unique authors. The most prolific author presented 49 times during this 19-year period. There were 28 authors who had 19 or more presentations, averaging at least 1 presentation per year. The top 1%  of authors' names appeared 1,195 times; however, many of these top-presenting authors copresented with other top-presenting authors, so this number does not reflect 1,195 separate presentations.The most collaborative presentation had 26 authors. Two thousand one hundred ninety-three presentations  had just 1 author, 882 presentations (15%) had 2 authors, and 1,011 (17%) had 3 authors; 1,695 presentations (29%) had between 3 and 26 authors.One thousand ninety-three records included a hospital-related keyword in the Author field, which might suggest that hospital librarians authored or coauthored approximately 19% of the total content. An academic-related keyword was included in the Author field for 4,517 records, indicating that academic librarians authored or coauthored approximately 78% of MLA annual meeting content over this 19-year period. There were 888 records that did not include either an academic-related keyword or a hospital-related keyword. These 888 records represented government, public, and special libraries as well as unique or abbreviated names of academic or hospital libraries. For example, “UCLA” would not be captured in the academic result set, although records that contained “University of California, Los Angeles,” would have been captured. US states appeared 7,420 times in the dataset, indicating numerous collaborations between institutions in different states. The state that most often appeared at least once per record was Maryland, appearing in 501 records, followed by California, New York, Washington, Pennsylvania, Virginia, Michigan, North Carolina, Texas, Illinois, and Florida as the 10 most frequently appearing states. The only US territory that appeared was Guam, listed on 3 records. An interactive map of the results can be viewed online . Figure There were 438 presentations with at least 1 international author. Canada was the country that was most frequently represented, with 193 presentations. There were 85 presentations with authors from the United Kingdom, 38 from Taiwan, 22 from China, 22 from Japan, 19 from Australia, 11 from Nigeria, and 10 from the Netherlands. The countries with fewer than 10 presentations were: India (9 presentations); Israel (8 presentations); Qatar (7 presentations); Belgium, Finland, France, Mexico, Switzerland, and Zimbabwe (5 presentations each); Botswana and South Africa (4 presentations each); Brazil, Ireland, and Uganda (3 presentations each); Kenya, Malaysia, Norway, Romania, and Zambia (2 presentations each); and Antigua, Armenia, Costa Rica, Dominica, Estonia, Ethiopia, Germany, Guatemala, Italy, Latvia, Madagascar, Malawi, Mali, Mozambique, Panama, Rwanda, Spain, Turkey, and the United Arab Emirates (1 presentation each).The goal of the automated content analysis was to programmatically group keywords in the corpus of the past nineteen years of MLA annual meeting abstracts. The topic model generated groups of word lists, or “topics,” enabling the author to assign a category to each topic based on its highly relevant keywords. The LDA topic model's sixteen categories are identified in the following list. The order reflects the ordering in pyLDAvis' output .The words in parentheses that follow the topic names are the first five words in each topic from the topic model. Italicized words in the descriptions also come from the topic model.Events: This topic seemed to represent a variety of interpersonal events and the communication that went into successful events. It showed librarian involvement in professional organizations with terms like MLA and conference, as well as involvement in broader communities. Librarians planned, marketed, and hosted these events.Electronic resources: Terms like portal, interface, and software appeared to describe platforms, while create, develop, and design might imply librarian creative control or influence. Terms that represented educational aspects, like tutorial and guide, also appeared.Publications: This topic coherently described librarians' responsibility for journal publications and might indicate librarian research as well as scholarly communications support. Open and access appeared here. Bibliometric studies might also be represented with words like impact, analysis, compare, and report.Evidence-based practice: This topic clearly represented evidence-based practice, with librarians providing and presenting on information resources in a clinical setting . Terms like quality, competency, and evaluate seemed to indicate outcomes assessment following evidence-based practice interventions.Collections: Unlike topic #3, which appeared to specifically describe librarians' interest in journals, this topic represented collection management. Usage was a highly relevant term here, measured by the keywords change, number, and increase. Purchase and cost also appeared.Academic instruction: This topic represented instruction, teaching, and learning in a specifically academic context, evidenced by terms like college, university, and program. Librarians helped learners develop literacy and skills, fulfilling competencies and enabling them to complete their assignments.Librarian roles and relationships: This topic showed how librarians perceived their professional contexts. Hospital and academic appeared here as workplace descriptions. Nurse and nursing seemed to represent nurses as common partners of health sciences librarians. Liaison described many librarians' relationship with their communities. Librarians brought knowledge and experience to their community collaborations, including through committee participation.Technical systems: This topic described technical and software issues or problems, with librarians seeking to implement solutions and improvements. Staff, manage, management, and department might describe personnel. Library-specific systems like catalog, database, and electronic resource also appeared.Special collections: History, historical, archive, and unique appeared to represent special collections and efforts to document those collections digitally via description and metadata. Exhibit also indicated sharing initiatives. There were also terms that seemed to show large-scale concern for the value of medical special collections, such as national, world, and future.General instruction: Terms like module and workshop indicated that this topic described instruction more broadly than the academic instruction of topic #6 and might point frequently to an online mode of delivery. There was an emphasis on instructional design and assessment, with terms like feedback, evaluate, develop, and improve.Literature searching: This topic reflected expert searching with guideline and systematic reviews, as well as references to specific sources like MEDLINE and PubMed. Librarians conducted and performed searches, retrieved search results, and studied the search process.Surveys: Surveys and questionnaires, common library research methodologies, were highly relevant to this topic. The topic also included other qualitative approaches such as interviews. Terms like measure, assess, and determine implied motivation for librarian-led research.Research support: This topic appeared to include research data management and bioinformatics services. It described collaborative institutional partnerships, with librarians as informationists, team members working to secure or facilitate grants and funding.Community outreach: Terms like community, local, state, and national highlighted the geography of outreach, while training, resource, and access pointed to specific activities and needs. NLM, network, and region also appeared, indicating the work of the National Library of Medicine (NLM) and the National Network of Libraries of Medicine (NLM).Patient education: This topic seemed to describe information provision to patients, with librarians facilitating consumer health literacy. It also might describe patient education information intended for a physician and provider audience. Family, child, senior, and cancer indicated health information needs of specific populations.Library services: This topic related to libraries' mission to serve their users, patrons, and customers by providing space and services, including changing models of reference services .LDA's output is a list of fractions per abstract. Each fraction in the list represents 1 topic and the level of “aboutness” that the topic contributed to each abstract. Each topic contributes a percentage of content to each abstract, with the total amount of content being 100%.Abstracts were classified according to which topic contributed the highest percentage to each. The topic that appeared most often as the most dominant topic was community outreach. The 16 topics were normalized so that each year's total amount of topic content added up to 100%. Normalization enables insight into how much discussion around each topic happened per year, regardless of the number of abstracts presented that year. For example, the research support topic's total percentage of contribution  in the year 2002 was 10.4%, and in 2013, the sum of research support's total contribution had risen to 33.8%. However, 2013 happened to be the meeting with the highest number of total presentations in this 19-year period, with 471 abstracts, so this contribution percentage might not be representative of the proportional discussion of the research support topic in other years with fewer total abstracts. After normalization, research support had its largest proportionate appearance in 2019, with 8.1% of the abstract content, and its lowest proportionate appearance was in 2004, in which its proportional representation was only 5.1%. The minimum and maximum normalized results for each topic are shown in The number of paper presentations has been rising since 2017, with a corresponding decrease in the relative number of poster presentations. In 2019, for the first time since 2001, more papers were presented than posters. This reversal might be related to the submission deadline. Prior to 2019, recent years had a single deadline for both poster and paper submissions –29. In 2Given the typically higher number of poster presentations compared to papers, submitters may believe that they have a greater chance of having their presentation accepted as a poster, and with only one chance to submit, they might prefer to have their submission accepted as a poster rather than rejected as a paper. In 2019, the ability to submit a paper presentation and still have a later chance to resubmit the same abstract as a poster might have encouraged more total paper submissions. This is a positive development, especially for early career librarians who are building a professional reputation. This analysis found that although MLA has been a venue for thousands of unique presenters, a minority of highly prolific authors' names have appeared with disproportionate frequency. If MLA's annual meeting is to prioritize diverse voices, additional research should be done to identify members who are underrepresented at the annual meeting and study the effects of the submission process and other changes on submitters' demographics. While MLA's Medical Library Education Caucus has encouraged new member participation with the New Voices travel stipend and forum , additioIn 2019, presentations averaged more than 3 authors per paper. Some annual meeting themes were more explicitly collaborative, for example, “Connections: Bridging the Gaps” in 2008 or “Reflect & Connect” in 2010, which theoretically could have encouraged submission of collaborative projects in particular years. The mean number of authors did reach a high point for posters in 2009, with 3.5 authors per poster, corresponding with a meeting theme of “iFusions.” However, collaborative authorship of paper presentations has been increasing every year since 2014, regardless of meeting theme. This collaboration might reflect the growing scale and complexity of librarians' responsibilities. Information contexts are rapidly changing: with more users , 34 and The number of hospital librarians' presentations at MLA annual meetings have consistently been much lower than that of academic librarians. Lessick et al. found in 2016 that hospital librarians were significantly less likely than their health sciences academic counterparts to engage in research activities, present research at conferences, or write research articles . HoweverIn the United States, authorship seems to largely reflect population levels. The mid-Atlantic states display particularly high authorship. This region is currently covered by three MLA chapters—Mid-Atlantic Chapter, New York-New Jersey Chapter, and Philadelphia Regional Chapter—indicating a high concentration of health sciences librarians in those states, so correspondingly high meeting representation may be expected. Participation from states or territories with lower populations can be facilitated by targeted solicitation , scholarships for travel assistance, or acceptance of virtual presentations.Outside the United States, librarian participation remains relatively low. MLA occasionally holds joint meetings with international partners, such as the Canadian Health Libraries Association/Association des bibliothèques de la santé du Canada (CHLA/ABSC). Most of MLA's international authors in the study period come from Canada, likely due to the 2016 joint meeting with CHLA/ABSC and to Canada's geographic proximity to meetings in the United States. After Canada, the United Kingdom has contributed the largest number of the authors to MLA, followed by East Asian countries. There is a long tail of remaining countries in MLA's more than 400 internationally authored abstracts. Funding presents a challenge for international participants, although MLA offers a number of international scholarships to attend the annual meeting, as well as discounted membership rates to members from low-income countries [The topic results can be approximately grouped into two categories:“Looking out” reflects the people who are part of librarians' jobs, the communities we connect with, and the knowledge and skills we bring, teach, and develop on their behalf: librarian roles and relationships, events, community outreach, general and academic instruction, patient education, literature searching, evidence-based practice, and research support.“Looking in” involves the materials and processes that are part of our jobs and the ways we define, improve, and reinvent them: electronic resources, collections, special collections, publications, technical systems, library services, and surveys.With a few exceptions, the “looking out” group topics have peaked in relevance in the last 4 years. There are 3 exceptions to the recent higher relevance of “looking out” topics. The first is community outreach, which peaked in 2003. As seen in Another factor may be that, in the early 2000s, NNLM and the Regional Medical Libraries began to focus on consumer health . MLA conThe final exception to the “looking out” topics late peak trend is evidence-based practice, which peaked in 2007. This may be a slightly delayed culmination of the 1990s–2000s evidence-based practice paradigm shift in health care. Librarians have used the emergence of evidence-based practice to promote their expertise in searching for and organizing evidence and educating students and providers on finding and using the best quality evidence –45. Thouassess and determine, implying that a primary motivation of librarians who undertook research was to make improvements to systems and services. This aligned with Lessick et al.'s study of health sciences librarians, who considered research to be “very important” for “guidance in evaluating, improving, and initiating new library collections, services, and operations” [The “looking in” topics generally peaked in the early to middle years of the 2001–2019 period. The one exception was the surveys topic, which had its high point in 2019, indicating that librarians see their own research as more important than ever. Health sciences librarian–authored research articles have been increasing in number , so it mrations” .However, MLA content related to collections and operations has declined. Technical systems peaked at the beginning of this data set, in 2001. Its terms might imply that the late 1990s and early 2000s were a time of implementing new systems—such as websites, integrated library systems, online public access catalogs, link resolvers, and resource sharing networks—and MLA was used a venue to discuss this technological change. The library services topic also peaked early, in 2003. Its terms implied discussion about physical space and reference service, two traditional library provisions that were forever changed by new forms of digital communication and the shift to online collections. Special collections peaked in 2005, and the other collections-related topics peaked in 2011–2012. Although collection development remains a core activity of librarians and information management is highly valued by our communities , 48, thelibrary as an entity into an era defined by the many skills of librarians [Health sciences librarians in the early twenty-first century are spending more time at MLA annual meetings talking about “looking out” at their communities, and prioritizing library engagement through their relationships and visible services. This shift toward people-centered librarianship, enabled by technology that uncouples librarians from physical buildings and print collections, has been noted for some time. Funk's 2012 analysis found that over the last forty years of the twentieth century, librarians began exploring embedded methods and new ways of outreach to users, prioritized instruction, and embraced technological change . The 201brarians .This study provides additional perspective from librarians using MLA's annual meeting to discuss our current challenges and shape our profession's journey into a twenty-first century, librarian-defined future. As we develop and utilize our knowledge, skills, and expertise to build new relationships with our communities, update our resources and services, and invent new roles for ourselves in the future of health care, we must ensure that MLA and other meetings and conferences represent diverse voices of librarians doing diverse work.


Ambient recreational waters can act as both recipients and natural reservoirs for antimicrobial resistant (AMR) bacteria and antimicrobial resistant genes (ARGs), where they may persist and replicate. Contact with AMR bacteria and ARGs potentially puts recreators at risk, which can thus decrease their ability to fight infections. A variety of point and nonpoint sources, including contaminated wastewater effluents, runoff from animal feeding operations, and sewer overflow events, can contribute to environmental loading of AMR bacteria and ARGs. The overall goal of this article is to provide the state of the science related to recreational exposure and AMR, which has been an area of increasing interest. Specific objectives of the review include (1) a description of potential sources of antibiotics, AMR bacteria, and ARGs in recreational waters, as documented in the available literature; (2) a discussion of what is known about human recreational exposures to AMR bacteria and ARGs, using findings from health studies and exposure assessments; and (3) identification of knowledge gaps and future research needs. To better understand the dynamics related to AMR and associated recreational water risks, future research should focus on source contribution, fate and transport—across treatment and in the environment; human health risk assessment; and standardized methods.  Staphylococcus aureus (MRSA) compared to people infected with a sensitive form of the bacteria [The increasing rise of antimicrobial resistance (AMR) is one of the greatest threats to human health in the 21st century . Today, bacteria . As pathbacteria ,8. BecauAn increasing body of research identifies the environment as not only a recipient of drug-resistant bacteria, but as a reservoir and source of resistance genes ,11,12,13Escherichia coli (E. coli) and enterococci, and to bacterial pathogens are of interest in ambient recreational waters. Bacterial pathogens, include but are not limited to, those in the Enterobacteriaceae family  and Staphylococcus aureus , which have all been detected in ambient waters [Resistance to traditional fecal indicator bacteria (FIB), such as t waters ,18,19,20t waters ,29,30,31t waters ,32. The t waters ,28,31,33Both point and nonpoint sources of fecal contamination, and even naturally occurring bacteria, are potential origins of AMR bacteria and ARGs in recreational waters ,34,35,36The goal of this review article is to provide the state of the science related to recreational water exposure and AMR bacteria and ARGs. Objectives of this review are to (1) describe potential sources of antibiotics, AMR bacteria, and ARGs in recreational waters as documented in the available literature; (2) discuss what is known about human recreational exposures to AMR bacteria and ARGs using findings from epidemiological studies and exposure assessments; and (3) identify knowledge gaps and future research needs.Antibiotic resistance and resistance genes are intrinsic microbial attributes that existed long before the first antibiotic was utilized by humans in the early 1900s ,44,45,46Bacteria have proven adept at developing resistance to antibiotics produced by humans ,55. HumaWhile resistance genes provide adaptive advantages to bacteria when antibiotic exposure does occur, ARGs can also be carried and shared without the selective pressure of antibiotic exposure. For example, ARGs can be transferred within a bacterial population when maintenance of the gene does not affect a cell’s ability to thrive in its environment . AdditioAeromonas spp., which can be pathogenic and indigenous, may play a role in disseminating ARGs because they (1) naturally produce three β-lactamases; (2) contain mobile genetic elements that can be transferred to other human pathogens [Aeromonas spp. as a surface water indicator of ARGs when fecal contamination is not expected in environmental samples. Berendonk et al. [E. coli and enterococci as surface water indicators of ARGs because they are currently used to monitor microbiological water quality in recreational waters and are well characterized in terms of acquired antibiotic resistance [E. coli during recreational activities can be associated with gut colonization by these bacteria [E. coli have been isolated from recreational water in the United Kingdom, and gut colonization with ESBL-E. coli was confirmed among surfers recreating in those waters. Resistant genes can move between pathogenic and nonpathogenic microorganisms ,72, and athogens ; and (3)athogens ,75,76,77sistance ,78,79,80bacteria ,17,81. FPoint and nonpoint sources contribute to the presence and proliferation of AMR bacteria and ARGs in surface waters ,34,35,36E. coli discharged from combined sewer overflows using the abundance of AMR, E. coli concentrations, and the flow rate of combined sewage and secondary treatment effluents. Annual concentrations of AMR E. coli discharged from combined sewer overflows were approximately 5000 times higher than from WWTP effluent [Much of the research in the emerging field of environmental AMR has focused on wastewater as a source of AMR bacteria and ARGs to water. Untreated wastewater influent (or raw sewage) contains concentrated levels of pathogenic and nonpathogenic microbes ,84 and aeffluent . Additioeffluent ,92,93. Beffluent . There have been many studies evaluating AMR in raw sewage (or influent) of residential areas, hospitals, and municipal WWTPs ,95,96,97The occurrence of resistance genes in raw wastewater can vary both geographically and temporally. Pärnänen et al.  studied Originating in hospitals and other healthcare facilities, pathogenic bacteria with resistance against all or almost all existing antibiotic treatments are of increasing concern . AntibioEnterobacteriaceae (CRE) and ESBL-resistance genes. For this study, the level of onsite treatment prior to discharge was not reported. The study also found a positive association between levels of ESBL-resistant bacteria in hospital wastewater and the relative abundance of ESBL-resistance genes, validating the assumption that more genes can lead to more phenotypically resistant bacteria [Divyashree et al.  analyzedbacteria . The sambacteria . Similarbacteria  reportedP. aeruginosa and ESBL-producing Klebsiella spp., Enterobacter spp., and Citrobacter spp. Their results highlight the role of clinical wastewater in the dissemination and development of multidrug resistance [Healthcare wastewaters also contain a higher-than-average concentration of pathogenic bacteria, which may share or acquire genes (via HGT) or be selected  during wastewater pre-treatment processes ,108. Thesistance . Pharmaceutical production facilities are another source of antibiotics in both raw sewage and surface waters and can contribute to AMR. In a national study of twenty U.S. WWTPs, influents impacted by pharmaceutical production facilities had significantly higher concentrations of 33 pharmaceuticals (of 120 tested) compared to control WWTPs not impacted by pharmaceutical production facilities . The conE. coli, to aquatic environments [Treated WWTP effluent may act as a point source of AMR once discharged to surface waters ,114,115.ronments ,120,121.Studies evaluating the prevalence of AMR in WWTP effluent indicate that wastewater treatment processes, including some secondary and tertiary treatment and disinfection processes, may not fully eliminate AMR bacteria and ARGs and that surviving bacteria retain their resistance properties ,128,129.In the U.S., municipal waste is generally treated by primary and secondary wastewater treatment processes, with many plants employing additional disinfection steps to remove pathogens before discharging the effluent into surface waters. Depending on the designated uses of the receiving water bodies, some U.S. WWTPs disinfect effluent seasonally, rather than year-round . PrimarySome studies have shown a relative increase in AMR bacteria and ARGs in treated effluent as compared to influent ,41,127. A WWTP that utilized advanced treatment processes  was identified as a point source of ARGs into surface waters in Duluth-Superior Harbor . A statiE. coli than effluent from WWTPs absent of wastewater from healthcare facilities [Drug-resistant microbes can spread into surface water and from there into recreational waters through insufficient treatment of hospital wastes . Reinthacilities , a findicilities . Effluencilities . High lecilities . Comparicilities . The concilities .In the U.S., most healthcare facilities discharge to municipal waste systems and are categorized as indirect dischargers, meaning waste is treated at a WWTP prior to entering surface waters . Of the Biosolids, the sludge generated by the treatment of sewage at WWTPs, are used as a soil amendment for agriculture, landscape, and for use in home gardens . They maE. coli, MRSA, and vancomycin-resistant Enterococcus spp. (VRE) in biosolids [Regarding land application of biosolids, Pepper and colleagues  reviewediosolids ,150,151.iosolids ,151. Runoff from rainfall can carry excess contaminants from biosolids to surface waters, including those used for recreation. Proximity to surface water is an important consideration when applying biosolids . More daAntimicrobials are used in agriculture, including animal husbandry and plant crops, and aquaculture to treat and prevent diseases and to promote growth and feed efficiency . GloballAnimal feeding operations (AFOs) and confined animal feeding operations (CAFOs) are a large source of the antibiotics and it is estimated that between 60 and 80 percent of livestock and poultry routinely receive antimicrobials . The preCampylobacter jejuni and Campylobacter coli was assessed in a survey of 200 cecal samples of chickens ready for slaughter from 20 major producers in Australia [C. jejuni isolates, the most commonly detected resistance was to tetracycline (n = 24) followed by resistance to the quinolones, ciprofloxacin (n = 16) and nalidixic acid (n = 16) [C. coli isolates exhibited less overall AMR. However, 5 of the 96 isolates were resistant to ciprofloxacin, nalidixic acid, azithromycin, erythromycin, and clindamycin, respectively, 4 isolates were resistant to telithromycin, and 3 isolates were resistant to tetracycline [E. coli, and enterococci, with 119 violations of NPDES permits between January 2016 and June 2018 (note that EPA’s Meat and Poultry products Effluent Guidelines are incorporated into NPDES permits: https://www.epa.gov/eg/meat-and-poultry-products-effluent-guidelines) [The animal supply chain also includes slaughter, rendering, and carcass disposal steps of the animal husbandry process, and bacteria from these sources are known to enter ambient waters. AMR of ustralia . Of the (n = 16) . C. coliacycline . Additioacycline ,173,174.delines) . The meddelines) . During delines) . The disdelines) . The amoErwinia amylovora, the fire blight pathogen, and Pseudomonas syringae, the cause of blister spots [Plant crops are potential sources of AMR because antimicrobial pesticides are utilized to disinfect or inhibit growth of unwanted microorganisms on crops and equipment ,177. Foler spots ,181,182.er spots . No signer spots . In a seAquaculture (or “aquafarming”) is frequently used in seafood production. In the U.S., there are over 2250 freshwater aquaculture farms and 875 saltwater aquaculture farms equaling over 249,000 and 214,000 surface acres, respectively . AquaculAntibiotics, other pharmaceuticals, and metal-containing products are used in aquaculture to prevent fouling and to feed and treat fish which leads to selection of antibiotic resistance in the aquatic environment . Up to 8Campylobacter spp., tetQ, and ermB) and land-applied biosolids (tetQ) [E. coli in wildlife fecal samples and report that ESBL-producing E. coli has been isolated from a variety of animals, including geese, ducks, deer, fox, owl, gulls, brown and Norway rats, and wild boar. Seagulls, birds, land mammals, cetaceans, and other wildlife may also be contributors of AMR bacteria and ARGs in the environment ,195,196.s (tetQ) . Birds as (tetQ) . For exas (tetQ)  revieweds (tetQ)  reviewedSalmonella spp. and HF183, respectively), with carriage rates associated with proximity to sewage outfalls [E. coli and enterococci and carry markers of AMR from human waste sites  to beaches, where they may be further dispersed [The role of wild birds and seagulls in the spread of AMR bacteria and ARGs warrants further evaluation due to their long-distance migration patterns . The fecoutfalls ,200. Seaispersed . E. coli were detected in 45 of 139 E. coli isolates from seagull feces collected at beaches in Portugal [E. coli isolates collected from two colonies of yellow-legged gulls in France [A study in California identified patterns in antibiotic resistance that aligned with time periods when bird feces were the main source of enterococci contamination in the surf zone . AnotherPortugal ; in 16 on France ; in 65 in France ; and 37 n France . S. aureus can potentially cause more severe health outcomes when drug-resistant strains  are present. MRSA is a growing public health problem, can be fatal, and has been found in ambient recreational waters in the U.S. For example, Plano et al. [S. aureus. A total of 1050 methicillin-sensitive S. aureus (MSSA) and 17 MRSA isolates were collected and analyzed from samples collected near bathers; 272 MSSA and 2 MRSA were isolated from ambient water samples [S. aureus in marine water. Additionally, Thapaliya et al. [S. aureus, including MRSA and MSSA. The authors indicate that further studies are necessary to draw public health conclusions.A variety of health endpoints are associated with exposure to fecal contamination in ambient waters, including gastrointestinal (GI) illness, urinary tract infections (UTIs), respiratory illnesses, and eye, ear, and skin infections. GI illness has been used as an index health endpoint in many recreational microbial risk assessments because it is often considered the most sensitive and prevalent health endpoint ,206,207.o et al.  collecte samples . The auta et al.  evaluateThe human health risks related to exposure to AMR bacteria and ARGs in recreational waters have been evaluated using epidemiological methods and risk assessment models. Though limited in number, epidemiological studies have evaluated the association between potential AMR bacteria exposure through recreational water activities and health endpoints such as GI illness  and gut In California, Griffith et al.  conducteE. coli. Surfers were significantly more likely to be carriers of cefotaxime-resistant E. coli (risk ratio (RR) = 2.95; 95% CI: 1.05–8.32) and blaCTX-M-bearing E. coli . The blaCTX-M genes represent nearly 80% of all ESBL-producing Enterobacteriaceae, which confer resistance to multiple antibiotics, such as fluoroquinolones, aminoglycosides, and tetracyclines and are characterized as a serious threat by the CDC’s 2012 and 2019 antibiotic resistance threats reports [In the United Kingdom, Leonard et al.  conducteEnterobacteriaceae. Recreational freshwater swimming within the past year was identified as one of several independent risk factors . The authors suggest that swimming may be a risk factor for intestinal colonization with E. coli with ESBL, and any subsequent UTI may be caused by a newly acquired ESBL-producing strain from the water. While this may highlight a possible connection between environmental pollution and resistant infections, the authors note that the link needs to be further investigated.Søraas et al.  conducteE. coli in recreational waters and in water in ditches surrounding poultry farms and municipal wastewater. Taking into consideration the effects of dilution and inactivation, the potential of ESBL-producing E. coli in source waters to reach downstream recreational waters was modeled, and the probability of human exposure through swimming was estimated. The authors suggest that exposure to ESBL-producing E. coli is possible when swimming in recreational waters located downstream of municipal WWTPs or livestock farms. However, the authors also note that the public health risk cannot yet be determined because the exposure is complex and more research is needed, for example, on colonization following asymptomatic carriage and HGT within the gut.Few studies have been conducted to estimate human health risks associated with AMR in recreational waters, with preliminary work focusing on exposure assessments. In the Netherlands, Schijven et al.  measuredE. coli resistant to third generation cephalosporins (3GCs) in coastal water samples and incorporated E. coli density data and reported volumes of water ingested during various water sports . Together, these data provided the mean number of 3GC-resistant ESBL-producing E. coli ingested during each water sport. Despite a low prevalence of 3GC-resistance in E. coli (0.12%), the authors conclude that there is still an identifiable human exposure risk for recreational water users. Based on their analysis, over 6.3 million water sport sessions in 2012 were estimated to have resulted in the ingestion of at least one 3GC-resistant E. coli. Since this study only considered resistance to 3GCs, it likely underestimates the true recreational exposure to AMR bacteria.Leonard et al.  estimateE. coli and also estimated human exposure to AMR bacteria in recreational water. The authors analyzed sequence data from E. coli metagenomes collected from 13 bathing water sites in England to determine the diversity and abundance of associated ARGs. The number of ARGs ingested by recreators was estimated using these sequence data, E. coli concentration data collected as part of The Environment Agency’s routine monitoring of 215 English bathing waters (samples collected weekly between mid-May and the end of September), and estimated water ingestion volumes associated with various recreational activities. E. coli in these bathing waters were found to harbor, on average, 1.24 ARGs per cell, and it was estimated that recreators ingested at least 100 E. coli-borne ARGs during each recreational water activity session [In a separate study, Leonard et al.  develope session .E. coli in two rivers located near different WWTPs and recreational beaches in central Italy. The authors conducted a field survey to better understand how much water was consumed by beachgoers and created a quantitative risk assessment model using dilution and decay rates of antibiotic resistant E. coli from the WWTP effluent into the river, dilution rates from the river into the swimming areas, and incidental water ingestion rates from swimming. The mean predicted human exposure levels  to antibiotic resistant E. coli were between 0 and 345 colony-forming units per 100 milliliters. Such modeling suggests that human exposure to resistance occurs through recreational water activities.O’Flaherty et al.  examinedThis paper summarized available information on possible sources of AMR bacteria and ARGs in ambient surface waters and human recreational exposures to AMR bacteria and ARGs using available human health studies. Regarding our understanding of recreational water exposures and AMR risks, this review exposed a number of knowledge gaps and research challenges including the following: (1) information on contributions on various sources to environmental AMR; (2) behavior of AMR bacteria and ARGs in the environment ; (3) human health risks associated with recreational exposure to AMR bacteria and ARGs in water; and (4) standardized methods for culturable and gene targets. Additional data on environmental sources of AMR bacteria and ARGs and their associated contributions are needed. Amarasiri et al.  providedA better characterization of potential sources and their respective contributions would help with quantifying exposure, risk assessment modeling and possible mitigation strategies. Specifically, more data are needed on the variability and load contributions of potential key sources of AMR bacteria and ARGs  in surface waters. For medical/health care facilities, a better understanding of their impact on municipal wastewater systems would inform future best practices and treatment needs. Active surveillance in surface waters of clinically relevant AMR bacteria strains is needed, as environmental surveillance could provide added protection of human, animal, and ecosystem health . HuijberDespite the environmental and public health significance of the well-known fecal contamination sources, there are data gaps regarding how AMR from these sources persist in the environment, across wastewater treatments, and what concentrations may pose a human health risk. A better understanding of how AMR bacteria and ARGs persist in water and wastewater would help WWTPs develop a more targeted approach to removing AMR bacteria and ARGs, thereby reducing their presence in effluent discharged into surface waters. Treatment processes are complex, and the effectiveness of disinfectants may be affected by a variety of factors such as dose, contact time, temperature, and pH ,131. AnoWhile the presence of AMR bacteria and ARGs in ambient surface water has been well-documented, their behavior in the environment and response to selection pressure is unclear ,33,214. The existing literature lacks a well-developed approach for the development of human health risk assessments of AMR bacteria and ARG exposure in recreational water. Risk assessments of how AMR bacteria and ARGs interact with environmental bacteria and threaten human populations are lacking ,33,85. AAdditionally, before a quantitative risk assessment can be conducted, there is a need for formal dose–response models with respect to specific pathogenic AMR bacteria ,30,32,33intI [mcr-1 [sul1 [tetW [blaTEM [No standard methods exist for measuring AMR in recreational waters. A general observation of the available literature is that researchers are using a variety of different methods to study environmental AMR with variations in the bacteria or genes targeted and the antibiotics tested, which precludes cross comparison of datasets. To develop a standardized method, researchers will likely need to decide whether to adopt culture-based methods, molecular methods, or both. For the molecular methods, there will also need to be consensus on the appropriate gene targets. Several gene targets appear promising as indicators of environmental AMR, including intI ,217, mcrI [mcr-1 ,219, sul-1 [sul1 ,221, tetl1 [tetW ,221, bla [blaTEM ,223, and [blaTEM ,224,225. [blaTEM . Some re [blaTEM ). The coE. coli may be useful as it is a standard water quality indicator and its evaluation would complement existing efforts for surveillance of AMR. In the U.S., resistant E. coli is monitored by all three federal agencies  collaborating in the National Antimicrobial Resistance Monitoring System (NARMS) aimed at monitoring trends in AMR from humans, animals, and retail meats [E. coli, specifically ESBL-E. coli, for global surveillance spanning human, animal, and environmental samples [-E. coli in recreational waters would ensure testing of viable organisms with human health relevance. Use of culture-based E. coli would also facilitate comparison of data for recreational waters and other types of samples across national and international databases.Regarding a culturable method for targeting AMR in recreational waters, il meats . Additio samples ,230,231.Addressing these research needs will help inform policies and practices to reduce the spread of AMR into the environment and protect recreational water users from exposure.As microbial resistance to antibiotics and multidrug resistant strains rise within the natural environment and clinical settings, improving antibiotic stewardship has become more pressing . When ev


Burkholderia pseudomallei, that is widespread throughout the tropics. It is estimated to affect approximately 165,000 people every year and kill 89,000 of them Melioidosis is a severe and frequently fatal disease caused by an environmental bacterium, EBioMedicine, Madden and colleagues report the development of a tool, which they call ARDaP, for detecting antimicrobial resistance (AMR) determinants in next generation sequencing (NGS) data for B. pseudomallei, having found shortcomings with existing tools for this purpose B. pseudomallei isolates studied, and accurately predicted three previously undescribed determinants. In comparison, other commonly-used tools such as CARD, ResFinder and AMRFinderPlus failed to find any of the clinically-relevant variants in B. pseudomallei, and found only the intrinsic resistance genes present in all B. pseudomallei strains.In this issue of B. pseudomallei was a strange choice of organism for this study, reflecting the prior research interests of the authors. Although it is a species that is intrinsically resistant to many antibiotics which has a large and highly plastic genome organised in two chromosomes, and exhibits frequent horizontal gene acquisition B. pseudomallei has rarely if ever been seen, and so even when resistance does develop it is an evolutionary dead end. In addition, whilst ARDaP may be an extremely efficient way of detecting AMR determinants in B. pseudomallei, it is unlikely to be applicable in the majority of clinical laboratories that serve the poor, rural populations in the developing world that are most likely to suffer from melioidosis, at least for the foreseeable future.In many ways, B. pseudomallei, may not be included any datasets used to test AMR tools. There are many existing tools for determining AMR from sequencing data B. pseudomallei resistance determinants required Madden et al. to develop a custom database for this species. To extend the use to other species will require substantial effort to curate catalogues of species-specific resistance variants, and to make those available to the scientific community, as has been done for other widely used AMR databases B. pseudomallei disease. Despite these limitations, ARDaP represents an important step towards incorporating AMR detection from NGS into routine clinical practice in the future.In fact, the most important aspect of the study of Madden et al. is the potential for this approach to be adapted for use with any bacterial species which, like The authors confirm sole responsibility for the conception and preparation of this invited Commentary.Dr. Dance reports personal fees from InBios, outside the submitted work; Dr. Batty has nothing to disclose.


Lathyrus armenus (Boiss. & Huet) which are endemic in Turkey, were studied. Flavonol glycosides  rhamnoside) were isolated by different chromatographic methods and identified by 1H, 13C NMR, as well as 2D NMR and Mass spectroscopy techniques from ethyl acetate and aqueous fractions of L. armenus’s methanolic extract. This is the first study about secondary metabolites of Turkish Lathyrus species.In the last five decades study on plant secondary metabolites have been increasing. Higher plants with a wide range of secondary metabolites have been very important in the search of new therapeutic agents. In this study secondary metabolites of  Lathyrus  is the largest genus in tribe Fabeae and exhibits an extensive distribution  were investigated, and the constituents were isolated and identified based on the spectral data. Generaln-hexane, CHCl3, EtOAC were purchased from Sigma- Aldrich. Silica gel 60 (0.063-0.200 mm), acetonitril and RP-modified Silica Plates were purchased from Merck. The ACE 5, C18 250×4.6 mm, and ACE 10, 250×10 mm HPLC column was used for analysis. Varian Mercury 400MHz Nuclear Magnetic Resonance was used for NMR spectroscopy and Agilent 1100 HPLC series was used for HPLC analysis. Methanol, Plant material and isolation processL. armenus was collected from Sivas, Turkey, during the flowering period. Voucher species were identified and the plant sample was deposited for future reference in Ankara University, Faculty of Pharmacy (AEF: 26680). L. armenus (800 g) were extracted with methanol by Soxhlet extraction for 12 h. The residue was evaporated under the vacuum and dried (156.185 g) and then partitioned successively between H2O and n-hexane (47.238 g), CHCl3 (1.440 g), EtOAC (5.810 g), the residue or water fraction (97.129 g). The dried and powdered aerial parts of 2O (100:13.5:10 V:V:V) solvent system to obtain 9 fractions. Sub-fraction 7 was re-chromatographed on reverse phase TLC plates to purify compound 1 (0.133 mg). The MeOH-H2O (7:3 V:V) solvent system was used and Rf value for this compound was 0.4. Water fractions (97.129 g) also were fractionated by column chromatography on silica gel using a solvent system composed of EtOAC:MeOH:H2O (100:13.5:10 V:V:V) and EtOAC:MeOH (50:50 V:V:V) resulting in 10 sub-fractions. Sub-fraction 6 was purified on a C18 semi preparative HPLC column by Acetonitril: H2O mixture to obtain 2 (6 mg), 3 (10 mg), 4 (12 mg), 5 (13.5 mg) compounds. The retention time of these compounds in semi preparative HPLC, were 17,17.3, 19, 21, min respectively. The structure of the isolated compounds has been established by using spectroscopic methods such as 1H, 13C NMR, as well as 2D NMR and Mass spectroscopyThe EtOAc fraction (5.810 g) was subjected to column chromatography over silica gel with EtOAC:MeOH:HL.armenus methanolic extract. In current study, 5 flavonoid derivatives were isolated for the first time from 1, yellow amorphous powder, isolated from ethyl acetate fraction, ESI-MS fragmentation displayed [M+H]+ m/z 287.14 ion peak, indicating the kaempferol aglycone and [M+H]+ m/z 595.32 ion peak for kaempferol glucosyl rhamnoside. The molecular formula was established as C27H30O15 (Kaempferol -3-O-glucosyl (1-2) rhamnoside).Compound 2 yellow amorphous powder, was isolated from water fraction, ESI-MS fragmentation displayed [M+H]+m/z 287.14 ion peak, indicating the kaempferol aglycone and [M+H]+m/z 741.13 ion peak kaempferol rhamninoside. The molecular formula was established as C34H42O20 (Kaempferol - 3-O-rhamninoside).Compound 3 a yellow amorphous powder, was isolated from water fraction, ESI-MS fragmentation displayed [M+H] + m/z 317.37 ion peak, indicating the rhamnetin aglycone, and [M+H]+m/z 770.71 ion peak rhamnetin rhamninoside. The molecular formula was established as C34H43O20 (Rhamnetin-3-O-rhamninoside).Compound  4 a yellow amorphous powder, was isolated from water fraction, ESI-MS fragmentation displayed [M+H]+ m/z 301.24 ion peak, indicating the rhamnocitrin aglycone, and [M+H]+m/z 754.35 ion peak rhamnocitrin rhamninoside. The molecular formula was established as C34H43O20 (Rhamnocitrin-3-O-rhamninoside). Compound5 a yellow amorphous powder, was isolated from water fraction, ESI-MS fragmentation displayed [M+H]+m/z 331.37 ion peak, indicating the rhamnazin aglycone, and [M+H]+m/z 785.40 ion peak rhamnazin rhamninoside. The molecular formula was established as C34H43O20 (Rhamnazin- 3-O-rhamninoside). The NMR spectral data measured in methanol-d are shown in Compound Lathyrus genus plants have endemic importance due to their usage as a food, fodder and ornamental crops. This species of plants contains approximately 25% of protein, and they are similar to other commonly used grain legumes, such as peas and fava beans. However, nonprotein amino acid ) also is found in Lathyrus species in low concentration. Literature data have demonstrated that Lathyrus species contain functional compounds including phenolics with antioxidant activity. The Lathyrus species it has been reported that quercetin, kaempferol, and their glycosides such as 3-O- glucosides, 3-rutinoside, 3-3-glucoside, 3,7- glucoside, 3-galactoside, 3-sophoroside, 7- glucoside, 3-robinobioside and 3-lathyroside-7-rhamnoside were isolated (Related to chemistry of isolated .L. cicera released that seeds contain 37 glycosylated flavonoids which were identified by HPLC-DAD-ESI/MS method. Kaempferol glycosides were detected as the main glycosides and quercetin, isorhamnetin, apigenin as well as luteolin glycosides were also determined (Further studies on termined .L. digitatus aerial parts content of phenolic acids and flavonoids were determined. Most of the flavonoids were identified as kaempferol and quercetin derivatives according to their mass fragmentations (ntations . KB activation, aldose reductase inhibition, insulin receptor activator, anti-carcinogenic, anti-mutagenic , etc. There are so many in-vivo and in-vitro studies about usefulness of flavonoids. They are using as nutraceutical, pharmaceutical, medicinal, and cosmetical pupose. (Lathyrus species. According to the litrature survey Flavonoids with 3-sophoroside-7-glucoside, 3-robinobioside 3-sophoroside and the 3-lathyroside-7-rhamnoside moiety were isolated from this genus. of Cicer, Lens, Pisum and Vicia indicate that each genus has a distinctive flavonoid profile (Flavonoids are omnipresent in higher plants, and improve plant–microbe interactions, plant–animal interactions. Flavonoids prevalence in plants make them important components in the control of inflammation and cancer prevention . Flavono pupose. , 18, 19.O-rhamninoside, Rhamnetin-3-O-rhamninoside, Rhamnazin- 3-O-rhamninoside, Rhamnazin- 3-O-rhamninoside and Kaempferol-3-O-glucosyl (L. armenus. Kaempferol-3-O-glucosyl (L. armenus (The present paper describes NMR study and structure of Rhamnocitrin-3-glucosyl -2 rhamnoglucosyl -2 rhamno armenus . 1H, 13C- NMR, HMBC, HSQC, COSY, and TOCSY data of glycosides moiety is clearly indicated the presence of rhamninoside in 2,3,4,5 compounds and glucosyl (1. The obtained data was compared with literature and approved. 2,3,4,5 compounds were isolated from Rhamnus species (1 was isolated from Ginkgo biloba and Hymenophyllum crispatum ( species , 21 and rispatum ,23.Lathyrus species are used as food and nutrient supplements in traditional diets all over the world. These plants have high content of polyphenol and antioxidant compounds. In this study L. armenus was selected to screen for its flavonoid contents. Flavonoids with rhamninose and rhamnoside moiety were detected for the first time in Lathyrus genus. The best of our knowledge is searching for bioactive compounds from endemic species which is very important, climate and geografic conditions force plants to produce different secondary metabolites. Furture, studies should be performed to reseal Lathyrus genus chemistry in detail.


Photoresponsive functional materials have gained increasing attention due to their externally tunable properties. Molecular switches embedded in these materials enable the control of phenomena at the atomic level by light. Metal–organic frameworks (MOFs) provide a versatile platform to immobilize these photoresponsive units within defined molecular environments to optimize the intended functionality. For the application of these photoresponsive MOFs (pho‐MOFs), it is crucial to understand the influence of the switching state on the host–guest interaction. Therefore, we present a detailed insight into the impact of molecular switching on the intermolecular interactions. By performing atomistic simulations, we revealed that due to different interactions of the guest molecules with the two isomeric states of an azobenzene‐functionalized MOF, both the adsorption sites and the orientation of the molecules within the pores are modulated. By shedding light on the host–guest interaction, our study highlights the unique potential of pho‐MOFs to tailor molecular interaction by light. Light on interaction: Photoresponsive molecular switches embedded within metal–organic frameworks enable the modification of both the adsorption sites and the orientation of guest molecules. Atomistic simulations predict the high potential of these photoresponsive materials to change intermolecular interactions (see figure). Stimuli‐responsive molecular switches enable the transfer of external stimuli such as light into well‐defined molecular motion. Embedded in precise molecular environments, these stimuli‐responsive molecular units perform a variety of essential functions in biological systems.cis/trans), which differ distinctively in their physical and chemical properties.In this context, azobenzene has been one of the most employed molecular building units.4O tetrahedra corners in MOF‐5 by azobenzene derivatives , we show further that the effect of the switching unit depends on even minor structural modifications and differences of the guest molecules, opening the path to tailor multicomponent systems by light.Soft porous materials such as metal–organic frameworks (MOFs) enable, due to their tool‐kit assembly,trans isomer (transA) together with the energy differences (with respect to transA) of in total four stationary points of both cis and trans isomers, which differ mainly by the torsion around C5‐C4‐N6‐N7 . The derived classical model is able to describe both the structural and dynamic properties of the four isomers of the model in excellent agreement with the QM data . Going beyond the stationary points, we validated further our force field by calculating the energy profile connecting the transA/B conformers. Although the intermediate structures were not included in the fitting procedure, the energy data calculated by our parametrized forced are well in line with corresponding QM calculations  and 36° (B3LYP+GD3), respectively, see also Figures S2–S6, Supporting Information), a small deviation around 0° is found. However, since we are interested in calculating the host–guest interactions at 300 K, the low energy barrier between the two configurations around 0° of 4.3 kJ mol−1 at the DFT level can be readily be overcome during the molecular dynamics simulations and thus only play a minor role for the further analysis.The high potential of atomistic simulations applying classical force fields, which allow to cover large time and length scales, to advance the understanding of MOF‐based materials has been discussed extensively.The influence of the rotational degrees of freedom of the flexible units on the adsorption sites in MOFs has previously been investigated in different studies.all‐trans azo‐MOF‐5 point predominately into the B‐cells. In contrast, the free phenyl rings in all‐cis azo‐MOF‐5 can be considered as immobilized benzene molecules .Our atomistic simulations of the benzene loaded azo‐MOF‐5 indeed confirm this hypothesis, by revealing a change of the interaction of the benzene molecules with the primary adsorption site. In the . Figure  b. At thθ, which describes the orientation between the surface normal of the aromatic ring of the guest molecules and a vector representing the diagonal of the A‐cell nearest to the guest molecule . In contrast, inspecting the probability distribution of θ for mesitylene, a dependency with respect to the switching state is evidenced . Despite the preferred orientation for the mesitylene molecules at θ=30–60°, a distinct change in the probability distribution is observed between all‐cis and all‐trans azo‐MOF‐5. In all‐cis azo‐MOF‐5 the probability of molecules with θ=60–90° increases by a factor of almost two, marking the influence of the two switching states on the orientation of the mesitylene within the pho‐MOF. To get a detailed picture of the host–guest interaction, we dissected the orientation of each molecule with respect to its center of mass. The corresponding probability distribution illustrates a clear correlation between the adsorption sites and the orientation ; (ii) with both the inorganic unit and the organic linker (θ=30–60°) and (iii) primarily with the IBMs (θ=60–90°). This assignment is supported by the RDFs between the mesitylene molecules and the IBMs, which provide information about the linker–guest interaction at each ODAS  and 39° (B)). Note that the deviation compared to the reference models  and 56° (D), respectively, changes from 83:17 % for θ=0–30° to 75:25 % for θ=60–90°. The significantly different host–guest interactions for different ODAS depending on the photo‐switching state is further clarified by extracting the corresponding molecular configurations from our atomistic simulations  were equilibrated at 300 K for 0.5 ns using the Nose‐Hoover thermostat.The classical molecular mechanics calculations, including the Molecular Dynamic (MD) simulations, were performed using a locally modified version of the TINKER program package.All DFT calculations were done using the Gaussian 16 codeThe authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re‐organized for online delivery, but are not copy‐edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.SupplementaryClick here for additional data file.


Introduction: Khorana score (KS) stratifies patients into low, intermediate, and high risk groups for venous thromboembolism (VTE). We examined the generalizability of the KS to risk of VTE and association with mortality.Methods: A retrospective cohort study was conducted at Mount Auburn Hospital, Cambridge, Massachusetts. Patients aged 18 years or older undergoing chemotherapy were included. All patients were evaluated for a six-month period. Primary study endpoints were VTE or mortality.Results: Some 277 participants were included with a mean age of 63.95 . The incidence proportion was 6.13% and a total of 17 VTE events were reported over a 2.5-year period. Compared to those with a low KS (0), those with a high KS (3 or above) had 6.4 times (p=0.032) while with an intermediate KS (1-2) had 2.6 times the odds of having a VTE event (p=0.22).Those who had a VTE had 4.03 times the odds of death compared to those who did not have a VTE (p=0.006). Compared to those with a low KS, those with a high KS had 5.7 times (p=0.02) the odds of six-month mortality and 5.04 odds (p=0.001) of mortality at any time.Conclusion: High KS was associated with increased odds of VTE and mortality in our study. Venous thromboembolism (VTE) is one of the major causes of morbidity, and the second most frequent cause of death in a cancer population -2. CanceAdditional factors also contribute to a greater risk of VTE occurrence among cancer patients, such as increased BMI, increased platelet count, increased leukocyte count, and diminished hemoglobin levels . Also ofTo date the KS and other scoring systems for high risk of VTE have been assessed and studied at major research centers, but the majority of the US population seeks care in community setting hospitals. The prevalence and risk for thrombotic events may be different in this group of patients as in general they may be older, less mobile, and have higher rates of co-morbidities than patients seeking care at a major research center. In this study we assessed the generalizability of the KS to risk of VTE in a community hospital and determined association with mortality.A retrospective cohort study was conducted at the outpatient Hematology and Oncology clinic of Mount Auburn Hospital, Cambridge, Massachusetts. Patients 18 years or older undergoing chemotherapy for a newly diagnosed cancer or new chemotherapy treatment after disease progression/re-occurrence with at least three months of lapsed time from prior chemotherapy were included in this study. The Khorana predictive model for cancer and chemotherapy-associated VTE was used to evaluate the risk of developing a VTE . ExclusiPatients were evaluated for a six-month period beginning from January 2016 to December 2017 for VTE events. De-identified data collected during the study included patient characteristics , past medical history , characteristics of cancer . The outcomes we recorded were incident VTE, pulmonary embolism (PE), and death. Ultrasound dopplers or CT (PE Protocol) was used to confirm a diagnosis of VTE.Data were collected and analyzed using STATA . Simple logistic regression, chi-square, and frequencies were used for analysis. Primary study endpoints included VTE or mortality at six months. We also continued to follow the patients and their mortality was documented anytime during the subsequent 2.5-year study period. This study was approved by the Internal Review Board at Mount Auburn Hospital.Demographic characteristicsA total of 339 patients were screened, of which 277 were found to meet the eligibility criteria. The mean age was 63.95 SD ± 12.47) with a median of 65 years (25-89). Detailed demographics are presented in Table 2.47 withA total of 17 VTE events (6.1%) were observed during the study period, of which four were PE and the remaining events were classified as venous embolism of extremities or abdominal vasculature. Stage 1 cancer patients had no VTE events while stages 2-4 were associated with 3, 4, and 10 events respectively. Further findings are discussed in Table Of the 62 patients excluded from the study, five had a VTE event in less than three months, 26 had prior chemotherapy in the last three months, while 35 had a known comorbidity for which they were already on anticoagulation.An increase in KS was associated with VTECompared to those with a low KS (0), those with a high KS (3 or above) were more likely to have a VTE event . The difference between low and intermediate KS (1-2) was not significant . Patients with metastatic cancer had higher likelihood of having a VTE event than those without metastatic cancer, though this was not statistically significant . The risk of VTE tended to be higher with aspirin use compared to patients not using aspirin, though this difference did not reach statistical significance .VTE and KS were associated with deathDuring the entire study period of 2.5 years, VTE was associated with an increased risk of death . Compared to those with a low KS, those with a high KS were at greater risk of death in the first six months of follow-up , and mortality at any time in the study . Intermediate KS also showed increased mortality which was statistically significant during the overall study period  but not significant at a six month interval . Compared to those with a low KS, those with a high KS had 5.04 times the odds of mortality, adjusting for cancer stage (metastatic vs. nonmetastatic) . Compared to those with a low KS, those with an intermediate KS had 2.54 times the odds of mortality, adjusting for cancer stage . Findings remained unchanged even after adjusting for age and sex.Our study reports that even in a community hospital, high KS is associated with increased odds of VTE as well as mortality. The occurrence of VTE, either in the form of deep venous thrombosis (DVT) or PE, leads to significant morbidity and mortality in cancer patients. Of the 500,000 new cases of VTE diagnosed annually in the United States, approximately 20% are associated with malignancy . VTE amoIn the 277 patients we studied, the cumulative incidence of VTE was 6.13%, which is consistent with other published studies -13. The While multiple studies have validated the KS as a predictive tool for chemotherapy-associated VTE, there are some studies that say otherwise , 14-18. Consistent with other published literature, our study reported that patients with VTE, compared to no VTE, were at significantly increased risk of mortality. Kuderer et al. in their study reported that VTE was associated with significantly worse prognosis than patients without VTE . Based oOur observation that a majority of our study patients were on aspirin (76.9%), but there was a trend towards an increased risk of having VTE with aspirin is hypothesis-generating and warrants further exploration in other cohorts. This observation is in line with the American Society of Clinical Oncology’s current guidelines against using aspirin for thromboprophylaxis . DespiteThough this study gives an insight into the real-world experience of using KS, it also has several limitations. This is a small retrospective single center study. We were limited by the small sample size which may account for the large CI and lack of precision in the OR estimates and reliability of the results. The effect of specific chemotherapies was not evaluated or controlled for. Our institute caters to higher proportion of Caucasian elderly population and findings might not be applicable to racially diverse younger population.In this study high KS was associated with increased odds of VTE and mortality after adjusting for cancer stage. High KS and intermediate KS were also statistically associated with increased mortality at any time. Aspirin use showed no protective effect and was associated with increased odds of VTE events though not statistically significant.


Daphnia magna to both dopamine and the dopamine reuptake inhibitor bupropion, an antidepressant that enters aquatic environments via various pathways. We recorded a range of life-history traits across two food levels. Both treatments induced changes to the life-history reaction norm slopes. These were due to the effects of the treatments being more pronounced at restricted food ration, where controls had lower somatic growth rates, higher age and larger size at maturation. This translated into a higher population growth rate (r) of dopamine and bupropion treatments when food was restricted. Our findings show that the dopamine system is an important regulatory mechanism underlying life-history trait responses to food abundance and that bupropion can strongly influence the life history of aquatic species such as D. magna. We discuss why D. magna do not evolve towards higher endogenous dopamine levels despite the apparent fitness benefits.Expression of adaptive reaction norms of life-history traits to spatio-temporal variation in food availability is crucial for individual fitness. Yet little is known about the neural signalling mechanisms underlying these reaction norms. Previous studies suggest a role for the dopamine system in regulating behavioural and morphological responses to food across a wide range of taxa. We tested whether this neural signalling system also regulates life-history reaction norms by exposing the zooplankton  UnderC. elegans, dopamine is released from dopaminergic neurons when food is present [Apis mellifera carnica) and Drosophila larvae, dopamine is involved in learning to associate food odour with aversiveness of taste, and thereby mediates an avoidance behaviour towards toxic and/or unpalatable food [Strongylocentrotus purpuratus), dopamine reduces food acquisition through a shortening in arm length when food is abundant, which preserves energy that can be allocated to other functions [At the molecular level, the responses to environmental stimuli that produce these reaction norms are mediated by neural signalling mechanisms. Thus, knowledge about these mechanisms is important to understand how organisms adjust their phenotypes under environmental change , as well as directly affect their survival [Insights into the mediation of reaction norms by neurotransmitters are also of potential value for environmental risk assessment of pharmaceutical products. Specifically, in aquatic biota, neurotransmitter systems can be directly altered by anthropogenic activity through environmental release of antidepressants. Following administration to humans, antidepressants can be eliminated unmetabolized or as active metabolites and enter the aquatic environment through wastewater . Anothersurvival ,29. HencDaphnia magna in response to high versus restricted food ration. Daphnia are keystone zooplankton in freshwater ecosystems and model organisms for studying anthropogenic and natural stressors in these ecosystems [D. magna with natural dopamine levels will have life-history reaction norms that approach optimality with respect to fitness in response to food abundance, and that disruption of these levels will lead to a change in the response to food abundance and hence the slopes of these reaction norms. Furthermore, bupropion administration causes an increase in extracellular dopamine in the brain [In this study, we experimentally tested for the effects of dopamine and bupropion exposure on the reaction norms of life-history traits of osystems . They haosystems ,32. We hhe brain . Thus, i2.(a)D. magna were collected in November 2014, in a pond at Værøy Island , northern Norway. Ephippial eggs were hatched in the laboratory and propagated clonally. For this experiment, juveniles of a single clone (clone 47) of D. magna were asexually propagated for four successive generations prior to use. A maximum of 30 individuals of D. magna were cultured in 2.5 l aquaria at 20°C in a modified Aachener Daphnien Medium (ADaM) [2 concentration reduced by 50%), under long photoperiods (16 h L : 8 h D) using white fluorescent lamps. The medium was exchanged weekly and the animals were fed three times a week with Shellfish Diet 1800 (Reed Mariculture Inc.) at a final concentration of 3.2 × 105 cells ml−1.Ephippia containing resting eggs resulting from sexual reproduction of m (ADaM)  (SeO2 co(b)−1) was chosen for successfully inducing changes in D. magna growth based on a study by Weiss et al. [−1) was selected for being an environmentally relevant concentration that can be expected to influence life-history traits based on a pilot study we conducted prior to this experiment (see [−1) were prepared by dissolving bupropion hydrochloride  in ultrapure water . The stock solutions were then added to ADaM to create the desired bupropion exposure concentration. For the dopamine treatment, dopamine hydrochloride  was first dissolved in 100 ml ultrapure water before dilution in ADaM to the desired exposure concentration. Controls containing only ADaM medium were performed parallel to the exposure replicates.A full factorial design with control, dopamine, bupropion and two food rations (high versus restricted) was used, with thirty 50 ml replicate tubes for each of the six combinations . Aqueous exposure to dopamine allows us to directly manipulate this compound in the experimental organisms. The exposure concentration of dopamine  three times a week, and the animals were fed at each renewal event with Shellfish Diet 1800 at a final concentration of 2.88 × 105 cells ml−1 (ad lib at 20°C) for the high food ration and 8.6 × 104 cells ml−1 (30% ad lib at 20°C) for the restricted food ration. Day 0 marks the start of the experiment, which was completed when the last individual died. Male individuals (n = 9) and individuals that died from pipetting (n = 2) were removed and not replaced.For each replicate tube, a single female neonate (less than 24 h old) was introduced and kept at 20°C under long photoperiods (16 h L: 8 h D) until death. The medium was renewed in all replicates ((c)n = 27; nine samples collected in total from each of the dopamine, bupropion and control treatments). Simultaneously, the new exposure solutions and ADaM medium were sampled for bupropion and dopamine analysis .Conductivity (WTW LF 330 conductivity metre), pH (WTW pH 340i) and dissolved oxygen (WTW Multi 3410 multiprobe metre) were measured throughout the experiment, after medium renewal, in the new exposure solutions and ADaM medium used for the controls . Lower than expected concentrations of these compounds may have been caused by degradation during storage.The samples were stored at −20°C for a maximum of four months after collection, prior to analysis. Two complementary sample preparation protocols were employed to cover all concentration ranges: (i) dilute-and-shoot and (ii) liquid–liquid extraction. Subsequent analysis was performed by ultra-performance liquid chromatography coupled to a triple quadrupole mass analyser (UPLC-MS/MS). Further details on the method are provided in electronic supplementary material. Over the course of the experiment, pH, conductivity and dissolved oxygen were within the recommended range for testing of chemicals in idelines . The conet al. [2.72. Thereafter, individual replicates were checked daily to record age at maturation and age at second reproduction, defined as the time when eggs were first visible in the brood chamber. Body length at first reproduction was also measured using ImageJ as described above. Live progeny released were collected and counted to yield first and second clutch size. For each replicate, we measured the BL of three offspring that were randomly sampled from each of the first and second clutch. As a derived parameter, we calculated the first clutch biomass as the product of clutch size and average offspring DM for that clutch. Offspring from all subsequent clutches were removed at each medium change, and the longevity of the mothers was recorded.Immediately prior to exposure on day 0, neonates were photographed for body length measurements  using ImageJ v. 1.52a . BL measurements were then transformed to dry mass  using the equation by Yashchenko et al. : DM = 0.start is the dry mass (in mg) of the replicate at the neonatal stage, DMend is the dry mass (in mg) of the replicate at maturation and duration is the number of days between the two stages.The somatic growth rate (SGR) of each replicate was calculated using the equationr) was calculated based on the two first reproductive events from the Euler–Lotka equation,x can be either age at maturation or age at second reproduction, xl is the probability of survival to age x and xm is the average number of offspring produced by an individual of age x.The intrinsic population growth rate ((d)All statistical analyses and graphic illustrations were performed in R v. 3.5.2. . We firsnlme package was used to allow residual variance to differ among treatments and food (see [r), bootstrapped sample means were used to compute r values for which 95% confidence intervals were derived using the percentile method. Between-group differences in r were considered statistically significant in the case where 95% confidence intervals did not overlap. The models were implemented using the lme and gls functions in the package nlme [glm function in the package stats.Model selection followed a backwards selection procedure, where variables were removed sequentially, starting with random effects, using likelihood ratio tests . For GLSood (see  for an eood (see ). Poissoage nlme  and the p > 0.05 indicates that no significant paths are missing from the model and that it fits the dataset well [piecewiseSEM package [To determine the causal pathways from food ration to first clutch biomass through age and DM at maturation, we used confirmatory path analyses ,45. Becaset well . The pat package .3.(a)p < 0.001), whereas age at maturation, DM at maturation, DM of first and second clutch and age at second reproduction decreased when the resources became sufficient (p < 0.01) (figures p < 0.05 for bupropion treatment), whereas the opposite pattern was observed in the dopamine treatment (p < 0.001). Although the sign of the reaction norm (i.e. positive versus negative slope) did not depend on the exposure treatment for most traits, their steepness did . This was generally due to a more pronounced effect of dopamine and bupropion under restricted than under high food regimes . By contrast, at restricted food ration, the differences between control on one hand and dopamine and bupropion treatments on the other hand, became more pronounced (p < 0.01) . Moreover, exposure to dopamine and bupropion induced lower DM for first and second clutch compared to controls, independent of food level (p < 0.01) (r) at restricted food, this effect was steeper for the control than for the dopamine and bupropion treatments (p < 0.05). Under restricted food ration, r was 159% and 114% higher in the dopamine and bupropion treatments than in the control, respectively .Specifically, at high food ration, treatment had no effect for SGR, age at maturation and age at second reproduction (ns), whereas a strong effect of dopamine treatment was observed for DM at first reproduction ((b)p = 1 for all groups). A high food ration favoured earlier maturation (p < 0.05). The direct effect of food on age was however much smaller in magnitude in the dopamine and bupropion treatments compared to the control  (p < 0.05 for dopamine and bupropion treatments), but this effect was of smaller magnitude  than its indirect effect (through age at maturation), which is obtained by multiplying the path coefficients  . DM at mp < 0.05). In the control and dopamine treatments, the direct effect of age at maturation on biomass was positive, whereas it was negative in the bupropion treatment . Furthermore, the direct effect of food on biomass was larger in magnitude than its indirect effect  compared to the control. A positive effect of dopamine upregulation on somatic growth rate was also seen in Weiss et al. , who suset al. ,61. Evidet al. ,63. Accoher size ,65.βdirect > 0 and βindirect < 0 for both treatments). The opposite was true in the bupropion treatment (βdirect < 0 and βindirect > 0). Moreover, direct allocation at restricted food ration was, given its magnitude, sufficient to offset the negative effect of delayed maturation on clutch biomass seen in the bupropion treatment. The negative effect of delayed maturation on clutch biomass in the bupropion treatment was unexpected, given the positive association between adult age, adult size and ultimately offspring size, and it could be due to physiological disruptions specific to bupropion's mode of action. Previous studies on aquatic animals have reported a variety of negative effects of bupropion exposure on reproductive physiology and development. One study showed bupropion negatively affecting the testicular morphology and reproductive physiology of adult male fathead minnows [Danio rerio) development, as well as a disruption of enzymatic activity related to energy production, movement and detoxification [Despite the similar effects of bupropion and dopamine treatments on life-history reaction norms to food abundance, path analyses identified differences in their resource allocation responses. Specifically, the relative importance of direct and indirect resource allocation (through age at maturation) to reproduction (first clutch biomass) changed according to food abundance across treatments. In the control and dopamine treatments, indirect resource allocation to reproduction increased at restricted food ration while direct allocation decreased ( minnows . Anotherfication . Findingr). At restricted food ration, both treatments caused an increase in population growth rate (r). Individuals in these treatments allocated more resources to maturation and reproduction, advancing the timing of reproduction, which resulted in faster rates of population growth compared to the control. This boost in fitness did not induce any apparent long-term costs as longevity did not differ significantly across treatments at restricted food. This is an important finding as both the principle of allocation [D. magna do not evolve towards higher endogenous dopamine levels. One potential explanation for this may be that population growth rate estimates based on the timing and fecundity of the first two clutches is not always an appropriate fitness measure [Regardless of their mode of action, aqueous dopamine and bupropion induced similar changes with respect to population growth rates  on fitness. Nonetheless, our findings emphasize the role of the dopamine system as regulator of trait responses to food abundance and demonstrate that low but environmentally relevant concentrations of bupropion can alter the life history of