Targeted mutagenesis in a human-parasitic nematode.

Parasitic nematodes infect over 1 billion people worldwide and cause some of the most common neglected tropical diseases. Despite their prevalence, our understanding of the biology of parasitic nematodes has been limited by the lack of tools for genetic intervention. In particular, it has not yet been possible to generate targeted gene disruptions and mutant phenotypes in any parasitic nematode. Here, we report the development of a method for introducing CRISPR-Cas9-mediated gene disruptions in the human-parasitic threadworm Strongyloides stercoralis. We disrupted the S. stercoralis twitchin gene unc-22, resulting in nematodes with severe motility defects. Ss-unc-22 mutations were resolved by homology-directed repair when a repair template was provided. Omission of a repair template resulted in deletions at the target locus. Ss-unc-22 mutations were heritable; we passed Ss-unc-22 mutants through a host and successfully recovered mutant progeny. Using a similar approach, we also disrupted the unc-22 gene of the rat-parasitic nematode Strongyloides ratti. Our results demonstrate the applicability of CRISPR-Cas9 to parasitic nematodes, and thereby enable future studies of gene function in these medically relevant but previously genetically intractable parasites

Supervised machine learning for the early prediction of acute respiratory distress syndrome (ARDS).

PURPOSE: Acute respiratory distress syndrome (ARDS) is a serious respiratory condition with high mortality and associated morbidity. The objective of this study is to develop and evaluate a novel application of gradient boosted tree models trained on patient health record data for the early prediction of ARDS. MATERIALS AND METHODS: 9919 patient encounters were retrospectively analyzed from the Medical Information Mart for Intensive Care III (MIMIC-III) data base. XGBoost gradient boosted tree models for early ARDS prediction were created using routinely collected clinical variables and numerical representations of radiology reports as inputs. XGBoost models were iteratively trained and validated using 10-fold cross validation. RESULTS: On a hold-out test set, algorithm classifiers attained area under the receiver operating characteristic curve (AUROC) values of 0.905 when tested for the detection of ARDS at onset and 0.827, 0.810, and 0.790 for the prediction of ARDS at 12-, 24-, and 48-h windows prior to onset, respectively. CONCLUSION: Supervised machine learning predictions may help predict patients with ARDS up to 48 h prior to onset

Role of Forkhead Box P3 in IFNγ-Mediated PD-L1 Expression and Bladder Cancer Epithelial-to-Mesenchymal Transition.

UNLABELLED: Antagonism of the PD-1/PD-L1 axis is a critical therapeutic strategy for patients with advanced bladder cancer. IFNγ functions as a key regulator of PD-L1 in both immune as well as cancer cells. Forkhead box P3 (FOXP3) is a transcription factor synonymous in T regulatory cell function but with increasingly described functions in cancer cells. Here, we investigated the relationship between FOXP3 and PD-L1 in bladder cancer. We showed that FOXP3 is critical in the ability for IFNγ to activate PD-L1 in bladder cancer cells. FOXP3 can bind to the PD-L1 promoter and induces a gene program that leads to regulation of multiple immune-related genes and genes involved in epithelial-to-mesenchymal transition (EMT). Using in vitro and in vivo human and murine models, we showed that FOXP3 can influence bladder cancer EMT as well as promote cancer metastases. Furthermore, FOXP3 may be a convergent factor for multiple activators of PD-L1, including the chemotherapeutic drug cisplatin. SIGNIFICANCE: Historically a key transcription factor driving T regulatory cell function, FOXP3 has an increasingly recognized role in cancer cells. In bladder cancer, we defined a novel mechanism whereby FOXP3 mediates the activation of the immune checkpoint PD-L1 by the cytokine IFNγ. We also showed that FOXP3 induces other immune checkpoints as well as genes involved in EMT, promoting immune resistance and cancer metastases

A mammalian methylation array for profiling methylation levels at conserved sequences

Infinium methylation arrays are not available for the vast majority of non-human mammals. Moreover, even if species-specific arrays were available, probe differences between them would confound cross-species comparisons. To address these challenges, we developed the mammalian methylation array, a single custom array that measures up to 36k CpGs per species that are well conserved across many mammalian species. We designed a set of probes that can tolerate specific cross-species mutations. We annotate the array in over 200 species and report CpG island status and chromatin states in select species. Calibration experiments demonstrate the high fidelity in humans, rats, and mice. The mammalian methylation array has several strengths: it applies to all mammalian species even those that have not yet been sequenced, it provides deep coverage of conserved cytosines facilitating the development of epigenetic biomarkers, and it increases the probability that biological insights gained in one species will translate to others

The role of stakeholders in creating societal value from coastal and ocean observations

The importance of stakeholder engagement in ocean observation and in particular the realization of economic and societal benefits is discussed, introducing a number of overarching principles such as the convergence on common goals, effective communication, co-production of information and knowledge and the need for innovation. A series of case studies examine the role of coordinating frameworks such as the United States’ Interagency Ocean Observing System (IOOS®), and the European Ocean Observing System (EOOS), public–private partnerships such as Project Azul and the Coastal Data Information Program (CDIP) and finally the role of the “third” or voluntary sector. The paper explores the value that stakeholder engagement can bring as well as making recommendations for the future

BHPR research: qualitative1. Complex reasoning determines patients' perception of outcome following foot surgery in rheumatoid arhtritis

Background: Foot surgery is common in patients with RA but research into surgical outcomes is limited and conceptually flawed as current outcome measures lack face validity: to date no one has asked patients what is important to them. This study aimed to determine which factors are important to patients when evaluating the success of foot surgery in RA Methods: Semi structured interviews of RA patients who had undergone foot surgery were conducted and transcribed verbatim. Thematic analysis of interviews was conducted to explore issues that were important to patients. Results: 11 RA patients (9 ♂, mean age 59, dis dur = 22yrs, mean of 3 yrs post op) with mixed experiences of foot surgery were interviewed. Patients interpreted outcome in respect to a multitude of factors, frequently positive change in one aspect contrasted with negative opinions about another. Overall, four major themes emerged. Function: Functional ability & participation in valued activities were very important to patients. Walking ability was a key concern but patients interpreted levels of activity in light of other aspects of their disease, reflecting on change in functional ability more than overall level. Positive feelings of improved mobility were often moderated by negative self perception ("I mean, I still walk like a waddling duck”). Appearance: Appearance was important to almost all patients but perhaps the most complex theme of all. Physical appearance, foot shape, and footwear were closely interlinked, yet patients saw these as distinct separate concepts. Patients need to legitimize these feelings was clear and they frequently entered into a defensive repertoire ("it's not cosmetic surgery; it's something that's more important than that, you know?”). Clinician opinion: Surgeons' post operative evaluation of the procedure was very influential. The impact of this appraisal continued to affect patients' lasting impression irrespective of how the outcome compared to their initial goals ("when he'd done it ... he said that hasn't worked as good as he'd wanted to ... but the pain has gone”). Pain: Whilst pain was important to almost all patients, it appeared to be less important than the other themes. Pain was predominately raised when it influenced other themes, such as function; many still felt the need to legitimize their foot pain in order for health professionals to take it seriously ("in the end I went to my GP because it had happened a few times and I went to an orthopaedic surgeon who was quite dismissive of it, it was like what are you complaining about”). Conclusions: Patients interpret the outcome of foot surgery using a multitude of interrelated factors, particularly functional ability, appearance and surgeons' appraisal of the procedure. While pain was often noted, this appeared less important than other factors in the overall outcome of the surgery. Future research into foot surgery should incorporate the complexity of how patients determine their outcome Disclosure statement: All authors have declared no conflicts of interes

The Fool of the Family: Nativism and the Ku Klux Klan in Oregon's 1922 Election

77 pages. A thesis presented to the Department of History and the Clark Honors College of the University of Oregon in partial fulfillment of the requirements for degree of Bachelor of Arts, Spring 2014.This thesis examines the intersection of national nativist trends with local peculiarities in Oregon's 1922 election. A surge of nativism in the United States during the post World War I period aided in the rapid expansion of the Second Ku Klux Klan, which came to be the prevailing political force in Oregon in the early 1920s. The KKK did not bring nativism to Oregon; rather, the state's unique social and political culture, coupled with the refusal of sitting Governor, Ben W. Olcott to recognize the Klan as a threat to social harmony, aided in the achievement of unprecedented Klan success in Oregon. The scope of this success is witnessed in the 1922 election, which resulted in a victory for a Klan backed governor, Walter M. Pierce, and the passage of the nation's first Compulsory Education Bill. To understand the prevailing sentiments before, during, and after the election this thesis analyzes the correspondence and speeches of both governors, as well as contemporary newspapers. As Oregon attempted to answer questions of the relationship between state and identity plaguing the nation it took actions that retain significance not only for Oregonians, but all Americans

Show Entry: Little Peter Cotton Tail

Little Peter Cotton Tail by Amy Zelasco and Emily Pellegrini was an entry in the Student Entry category of the 2014 Edible Book Festival. Amy and Emily were among many participating EIU students.https://thekeep.eiu.edu/ediblebook_2014/1031/thumbnail.jp

Show Entry: Little Peter Cotton Tail

Little Peter Cotton Tail by Amy Zelasco and Emily Pellegrini was an entry in the Student Entry category of the 2014 Edible Book Festival. Amy and Emily were among many participating EIU students.https://thekeep.eiu.edu/ediblebook_2014/1031/thumbnail.jp

Laterality in Motor Preference, Spatial Attention, and Auditory Processing

John Pellegrini, Associate Professor of Biology, and Emily Love, student researcher, received a $2,000 award from the 3M Small Scale Grant program to examine whether individuals tend to be left-hemisphere dominant for a variety of functions, or instead show a mixed pattern of laterality