Ex-vivo drug screening of surgically resected glioma stem cells to replace murine avatars and provide personalise cancer therapy for glioblastoma patients.

With diminishing returns and high clinical failure rates from traditional preclinical and animal-based drug discovery strategies, more emphasis is being placed on alternative drug discovery platforms. Ex vivo approaches represent a departure from both more traditional preclinical animal-based models and clinical-based strategies and aim to address intra-tumoural and inter-patient variability at an earlier stage of drug discovery. Additionally, these approaches could also offer precise treatment stratification for patients within a week of tumour resection in order to direct tailored therapy. One tumour group that could significantly benefit from such ex vivo approaches are high-grade gliomas, which exhibit extensive heterogeneity, cellular plasticity and therapy-resistant glioma stem cell (GSC) niches. Historic use of murine-based preclinical models for these tumours has largely failed to generate new therapies, resulting in relatively stagnant and unacceptable survival rates of around 12-15 months post-diagnosis over the last 50 years. The near universal use of DNA damaging chemoradiotherapy after surgical resection within standard-of-care (SoC) therapy regimens provides an opportunity to improve current treatments if we can identify efficient drug combinations in preclinical models that better reflect the complex inter-/intra-tumour heterogeneity, GSC plasticity and inherent DNA damage resistance mechanisms. We have therefore developed and optimised a high-throughput ex vivo drug screening platform; GliExP, which maintains GSC populations using immediately dissociated fresh surgical tissue. As a proof-of-concept for GliExP, we have optimised SoC therapy responses and screened 30+ small molecule therapeutics and preclinical compounds against tumours from 18 different patients, including multi-region spatial heterogeneity sampling from several individual tumours. Our data therefore provides a strong basis to build upon GliExP to incorporate combination-based oncology therapeutics in tandem with SoC therapies as an important preclinical alternative to murine models (reduction and replacement) to triage experimental therapeutics for clinical translation and deliver rapid identification of effective treatment strategies for individual gliomas.

Fourth-Year Medical Students' Perceptions of Vascular Surgery: Can We Improve the Pipeline?

BACKGROUND: Since their inception, Integrated Vascular Surgery Residency (IVSR) programs have expanded widely and attracted highly competitive medical students by offering a more focused approach to learning both open surgical and endovascular techniques. However, despite substantial modifications to the training paradigm, a shortage of vascular surgeons is still projected through 2050. We aimed to gather and analyze fourth-year medical students' knowledge and perceptions of vascular surgery (VS) to further inform strategies for recruiting future vascular surgeons. METHODS: We sent anonymous electronic questionnaires to fourth-year medical students at 7 allopathic and 3 osteopathic medical schools, with questions detailing demographics, specialty preferences, and exposure to and perceptions of VS. Descriptive statistics were obtained, and responses were compared between students applying to surgical specialties (SS) and nonsurgical specialties (NSS). RESULTS: Two hundred eleven of 1,764 (12%) participants responded (56% female). 56% reported VS exposure, most commonly during the third year. 64 (30%) planned to apply to SS. 57% of respondents reported knowledge of the management of vascular disease, and 56% understood procedures performed by vascular surgeons. Ranking the importance of factors in choosing specialties, SS selected "experiences gained during medical school rotations" (P < 0.05), "types and/or variety of treatment modalities used in this field" (P < 0.001), and "interest in the pathology or disease processes treated" (P < 0.05) as highest priorities. NSS preferred "lifestyle (work-life balance) as an attending" (P < 0.001). Only 7% of all respondents believed vascular surgeons have a good work-life balance, with a larger percentage of SS (P < 0.001) agreeing. Stratified by gender, female students rated "limited ability of childbirth during residency and/or postponement of family plans" (P < 0.05), "gender-related concerns, such as discrimination at work or unfair career possibilities" (P < 0.001), and "fear of unfair competition" (P < 0.05) as potential negative aspects of VS careers. 55% of respondents believed the IVSR makes VS more appealing. CONCLUSIONS: Medical students perceive poor quality of life and work-life balance as deterring factors to a career in VS. Opportunities exist to educate students on the pathologies treated, procedures performed, and attainable quality of life available in our field. We should also continue to develop recruitment strategies to stimulate student interest and increase early exposure in VS.

The ongoing impact of the Covid-19 pandemic on endometriosis patients: A survey of 1,089 UK patients.

Background: The impact of Covid-19 on endometriosis patients is under-researched. Endometriosis has significant psychosocial effects on patients. Moreover, the mainstay of diagnosis and treatment of endometriosis is elective surgery, impacted as a result of healthcare strain. Objective: To better understand the effect of the Covid-19 pandemic on endometriosis patients. Materials and Methods: An online survey sent to adult UK endometriosis patients between 27th August and 15th September 2021. The study received HRA and HCRW research ethic committee approval. Main outcome measures: Effects of the Covid-19 pandemic on endometriosis symptoms and surgery. Results: We received 1,089 survey responses. Respondents had a median age of 34, and 82.0% of respondents were white British. 18.8% of respondents reported a previous positive Covid-19 PCR test. 84.6% of patients had been double vaccinated at time of response. 20 patients reported Covid-related hospital admission, with 1 requiring intubation. Large numbers of patients (31.4-55.2%) reported worsening of endometriosis symptoms during the pandemic. 69.2% of respondents reported worsening of associated mental health symptoms. Whilst 44% of respondents had elective endometriosis surgery planned, the majority of operations were disrupted, and 18.7% of total respondents did not have a new surgery date. Conclusions: More research and support are needed for endometriosis patients as they wait longer for surgery. A holistic approach, encompassing mental health needs, may be particularly beneficial for patients. What is new?: This is the first survey examining the effects of Covid-19 on endometriosis patients including data beyond January 2021.

Exposure of American lobster (Homarus americanus) to the pesticide chlorpyrifos results in changes in gene expression.

Chlorpyrifos is an organophosphate that is currently used to reduce arthropod pests for the protection of agricultural crops. Coastal marine ecosystems may be exposed to agricultural pesticides via runoff and pesticide exposure can impact the health and survival of non-target species such as the American lobster (Homarus americanus). In the current study, the gene expression changes of H. americanus stage IV larvae were evaluated to understand the physiological mechanisms affected by exposure to sublethal concentrations of chlorpyrifos. After 48 h chlorpyrifos exposure, surviving lobsters were processed for Illumina RNA sequencing (RNA-seq). Genes of interest that showed significant changes using RNA-seq were verified using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). Analysis of RNA-seq and the confirmation of gene expression patterns via RT-qPCR found altered expression in genes related to stress response (glutathione peroxidase 3 and heat shock protein 60), hypoxia response (hairy, astakine 2, hemocyanin), moulting (cytochrome P450 307a1 and chitinase), and immunity (astakine 2) pathways. Changes to gene expression were most notable in lobsters exposed to 0.57 µg/L chlorpyrifos.

The American lobster genome reveals insights on longevity, neural, and immune adaptations.

The American lobster, Homarus americanus, is integral to marine ecosystems and supports an important commercial fishery. This iconic species also serves as a valuable model for deciphering neural networks controlling rhythmic motor patterns and olfaction. Here, we report a high-quality draft assembly of the H. americanus genome with 25,284 predicted gene models. Analysis of the neural gene complement revealed extraordinary development of the chemosensory machinery, including a profound diversification of ligand-gated ion channels and secretory molecules. The discovery of a novel class of chimeric receptors coupling pattern recognition and neurotransmitter binding suggests a deep integration between the neural and immune systems. A robust repertoire of genes involved in innate immunity, genome stability, cell survival, chemical defense, and cuticle formation represents a diversity of defense mechanisms essential to thrive in the benthic marine environment. Together, these unique evolutionary adaptations contribute to the longevity and ecological success of this long-lived benthic predator.

Array-based dynamic allele specific hybridization (Array-DASH): Optimization-free microarray processing for multiple simultaneous genomic assays.

We report proof-of-principle experiments regarding a dynamic microarray protocol enabling accurate and semi-quantitative DNA analysis for re-sequencing, fingerprinting and genotyping. Single-stranded target molecules hybridise to surface-bound probes during initial gradual cooling with high-fidelity. Real-time tracking of target denaturation (via fluorescence) during a 'dynamic' gradual heating phase permits 'melt-curve' analysis. The probe most closely matching the target sequence is identified based on the highest melting temperature. We demonstrated a >99% re-sequencing accuracy and a potential detection rate of 1% for SNPs. Experiments employing Hypericum ribosomal ITS regions and HIV genomes illustrated a reliable detection level of 5% plus simultaneous re-sequencing and genotyping. Such performance suggests a range of potential real-world applications involving rapid sequence interrogation, for example, in the Covid-19 pandemic. Guidance is offered towards the development of a commercial platform and dedicated software required to bring this technique into mainstream science.

Materials genes of heterogeneous catalysis from clean experiments and artificial intelligence.

ABSTRACT: The performance in heterogeneous catalysis is an example of a complex materials function, governed by an intricate interplay of several processes (e.g., the different surface chemical reactions, and the dynamic restructuring of the catalyst material at reaction conditions). Modeling the full catalytic progression via first-principles statistical mechanics is impractical, if not impossible. Instead, we show here how a tailored artificial-intelligence approach can be applied, even to a small number of materials, to model catalysis and determine the key descriptive parameters ("materials genes") reflecting the processes that trigger, facilitate, or hinder catalyst performance. We start from a consistent experimental set of "clean data," containing nine vanadium-based oxidation catalysts. These materials were synthesized, fully characterized, and tested according to standardized protocols. By applying the symbolic-regression SISSO approach, we identify correlations between the few most relevant materials properties and their reactivity. This approach highlights the underlying physicochemical processes, and accelerates catalyst design. IMPACT STATEMENT: Artificial intelligence (AI) accepts that there are relationships or correlations that cannot be expressed in terms of a closed mathematical form or an easy-to-do numerical simulation. For the function of materials, for example, catalysis, AI may well capture the behavior better than the theory of the past. However, currently the flexibility of AI comes together with a lack of interpretability, and AI can only predict aspects that were included in the training. The approach proposed and demonstrated in this IMPACT article is interpretable. It combines detailed experimental data (called "clean data") and symbolic regression for the identification of the key descriptive parameters (called "materials genes") that are correlated with the materials function. The approach demonstrated here for the catalytic oxidation of propane will accelerate the discovery of improved or novel materials while also enhancing physical understanding. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1557/s43577-021-00165-6.

SPI-Hub™: a gateway to scholarly publishing information.

BACKGROUND: Advances in the health sciences rely on sharing research and data through publication. As information professionals are often asked to contribute their knowledge to assist clinicians and researchers in selecting journals for publication, the authors recognized an opportunity to build a decision support tool, SPI-Hub: Scholarly Publishing Information Hub™, to capture the team's collective publishing industry knowledge, while carefully retaining the quality of service. CASE PRESENTATION: SPI-Hub's decision support functionality relies on a data framework that describes journal publication policies and practices through a newly designed metadata structure, the Knowledge Management Journal Record™. Metadata fields are populated through a semi-automated process that uses custom programming to access content from multiple sources. Each record includes 25 metadata fields representing best publishing practices. Currently, the database includes more than 24,000 health sciences journal records. To correctly capture the resources needed for both completion and future maintenance of the project, the team conducted an internal study to assess time requirements for completing records through different stages of automation. CONCLUSIONS: The journal decision support tool, SPI-Hub, provides an opportunity to assess publication practices by compiling data from a variety of sources in a single location. Automated and semi-automated approaches have effectively reduced the time needed for data collection. Through a comprehensive knowledge management framework and the incorporation of multiple quality points specific to each journal, SPI-Hub provides prospective users with both recommendations for publication and holistic assessment of the trustworthiness of journals in which to publish research and acquire trusted knowledge.

Effects of Sublethal Chlorpyrifos Exposure on Postlarval American Lobster (Homarus americanus).

The organophosphate pesticide chlorpyrifos has been introduced to the marine environment via adsorption to agricultural soil runoff or as spray drift. Chlorpyrifos affects the survival of some larval decapod crustaceans, but no data exist on the impacts to the American lobster, Homarus americanus. The purpose of the present study was to assess the levels at which chlorpyrifos affects the survival of postlarval H. americanus. Using acute saltwater exposures, the 24- and 48-h median lethal concentrations were established for stage IV H. americanus (1.56 and 1.33 µg/L, respectively). Movement, acetylcholinesterase activity, intermoult period, specific growth rate, and moult increment were measured during exposure to sublethal concentrations. Movement patterns were assessed to establish a 48-h median inhibition concentration for cessation of normal movement (0.66 µg/L). Acetylcholinesterase activity was found to be inhibited immediately post-exposure to 0.50, 0.57, and 0.82 µg/L chlorpyrifos but could be recovered within a period (9-15 d) in clean seawater. Sublethal growth effects of increased intermoult period, decreased specific growth rate, and decreased moult increment were observed during exposure to an environmentally relevant concentration (0.82 µg/L). The present study suggests that H. americanus stage IV larvae were marginally less sensitive to chlorpyrifos compared with other decapods and that acute lethality of H. americanus postlarvae is not likely to occur with chlorpyrifos concentrations previously reported from aquatic environments. Environ Toxicol Chem 2019;38:1294-1301. © 2019 SETAC.

Sex matters in massive parallel sequencing: Evidence for biases in genetic parameter estimation and investigation of sex determination systems.

Using massively parallel sequencing data from two species with different life history traits, American lobster (Homarus americanus) and Arctic Char (Salvelinus alpinus), we highlight how an unbalanced sex ratio in the samples and a few sex-linked markers may lead to false interpretations of population structure and thus to potentially erroneous management recommendations. Here, multivariate analyses revealed two genetic clusters separating samples by sex instead of by expected spatial variation: inshore and offshore locations in lobster, or east and west locations in Arctic Char. To further investigate this, we created several subsamples artificially varying the sex ratio in the inshore/offshore and east/west groups and then demonstrated that significant genetic differentiation could be observed despite panmixia in lobster, and that FST values were overestimated in Arctic Char. This pattern was due to 12 and 94 sex-linked markers driving differentiation for lobster and Arctic Char, respectively. Removing sex-linked markers led to nonsignificant genetic structure in lobster and a more accurate estimation of FST in Arctic Char. The locations of these markers and putative identities of genes containing or nearby the markers were determined using available transcriptomic and genomic data, and this provided new information related to sex determination in both species. Given that only 9.6% of all marine/diadromous population genomic studies to date have reported sex information, we urge researchers to collect and consider individual sex information. Sex information is therefore relevant for avoiding unexpected biases due to sex-linked markers as well as for improving our knowledge of sex determination systems in nonmodel species.

Potassium and Magnesium Supplementation Do Not Protect Against Atrial Fibrillation After Cardiac Operation: A Time-Matched Analysis.

BACKGROUND: Despite a lack of demonstrated efficacy, potassium and magnesium supplementation are commonly thought to prevent postoperative atrial fibrillation (POAF) after cardiac operation. Our aim was to evaluate the natural time course of electrolyte level changes after cardiac operation and their relation to POAF occurrence. METHODS: Data were reviewed from 2,041 adult patients without preoperative AF who underwent coronary artery bypass grafting, valve operation, or both between 2009 and 2013. In patients with POAF, the plasma potassium and magnesium levels nearest to the first AF onset time were compared with time-matched electrolyte levels in patients without AF. RESULTS: POAF occurred in 752 patients (36.8%). At the time of AF onset or the matched time point, patients with POAF had higher potassium (4.30 versus 4.21 mmol/L, p < 0.001) and magnesium (2.33 versus 2.16 mg/dL, p < 0.001) levels than controls. A stepwise increase in AF risk occurred with increasing potassium or magnesium quintile (p < 0.001). On multivariate logistic regression analysis, magnesium level was an independent predictor of POAF (odds ratio 4.26, p < 0.001), in addition to age, Caucasian race, preoperative ß-blocker use, valve operation, and postoperative pneumonia. Prophylactic potassium supplementation did not reduce the POAF rate (37% versus 37%, p = 0.813), whereas magnesium supplementation was associated with increased POAF (47% versus 36%, p = 0.005). CONCLUSIONS: Higher serum potassium and magnesium levels were associated with increased risk of POAF after cardiac operation. Potassium supplementation was not protective against POAF, and magnesium supplementation was even associated with increased POAF risk. These findings help explain the poor efficacy of electrolyte supplementation in POAF prophylaxis.

Genome-wide methylation profile following prenatal and postnatal dietary omega-3 fatty acid supplementation in pigs.

The objective of this study was to determine how prenatal and postnatal dietary omega-3 fatty acids alter white blood cell (leukocyte) DNA methylation of offspring. Fifteen gilts (n = 5 per treatment) were selected from one of three treatments: (i) control diet throughout gestation, lactation and nursery phase (CON); (ii) algal omega-3 fatty acid supplementation enriched in EPA and DHA (Gromega™ ) fed throughout gestation, lactation and nursery phase (Cn3); or (iii) Gromega™ supplementation maternally, during gestation and lactation only, and control diet during the nursery phase (Mn3). At 11 weeks of age and after 8 weeks of post-weaning nursery feeding, buffy coat genomic DNA was subjected to methyl CpG binding protein sequencing. The methylation enriched profile mapped to 26% of the porcine genome. On chromosome 4, a 27.7-kb differentially methylated region downstream of RUNX1T1 was hypomethylated in the Mn3 and Cn3 groups by 91.6% and 85.0% respectively compared to CON pigs. Conversely, hypermethylation was detected in intergenic regions of chromosomes 4 and 12. Regulatory impact factor and differential hubbing methods were used to identify pathways that were coordinately regulated by methylation due to feeding EPA and DHA during pregnancy. Despite limited ability to detect differential methylation, we describe methods that allow the identification of coordinated epigenetic regulation that could not otherwise be detected from subtle single locus changes in methylation. These data provide evidence of novel epigenetic regulation by maternal and early life supplementation of omega-3 fatty acids that may have implications to growth and inflammatory processes.

Seascape genomics provides evidence for thermal adaptation and current-mediated population structure in American lobster (Homarus americanus).

Investigating how environmental features shape the genetic structure of populations is crucial for understanding how they are potentially adapted to their habitats, as well as for sound management. In this study, we assessed the relative importance of spatial distribution, ocean currents and sea surface temperature (SST) on patterns of putatively neutral and adaptive genetic variation among American lobster from 19 locations using population differentiation (PD) approaches combined with environmental association (EA) analyses. First, PD approaches (using bayescan, arlequin and outflank) found 28 outlier SNPs putatively under divergent selection and 9770 neutral SNPs in common. Redundancy analysis revealed that spatial distribution, ocean current-mediated larval connectivity and SST explained 31.7% of the neutral genetic differentiation, with ocean currents driving the majority of this relationship (21.0%). After removing the influence of spatial distribution, no SST were significant for putatively neutral genetic variation whereas minimum annual SST still had a significant impact and explained 8.1% of the putatively adaptive genetic variation. Second, EA analyses (using Pearson correlation tests, bayescenv and lfmm) jointly identified seven SNPs as candidates for thermal adaptation. Covariation at these SNPs was assessed with a spatial multivariate analysis that highlighted a significant temperature association, after accounting for the influence of spatial distribution. Among the 505 candidate SNPs detected by at least one of the three approaches, we discovered three polymorphisms located in genes previously shown to play a role in thermal adaptation. Our results have implications for the management of the American lobster and provide a foundation on which to predict how this species will cope with climate change.

Aerococcus viridans var. homari: The presence of capsule and the relationship to virulence in American lobster (Homarus americanus).

The relationship between virulence and encapsulation of Aerococcus viridans var. homari was evaluated by growing virulent (Rabin's) and avirulent (ATCC 10400) strains under varying culture conditions, and during challenge trials. Changes in capsule thickness were monitored using a modified lysine-ruthenium red (LRR) fixation method and transmission electron microscopy. The virulent Rabin's strain possessed a prominent capsule of 0.252 µm±0.061 µm that was diminished by in vitro growth conditions to 0.206 µm±0.076 µm. The ATCC 10400 strain capsule thickness decreased from 0.157 µm±0.043 µm to 0.117 µm±0.043 µm after 10 in vitro passages. The virulent Rabin's strain capsule was significantly thicker than the avirulent ATCC 10400 strain under all growth conditions. Rabin's strain, regardless of pre-challenge growth conditions or dose (high dose 10(7) or low dose 10(2)), was able to kill lobsters in 7 days at 15°C. ATCC 10400 strain, regardless of pre-challenge growth conditions, killed lobster only at high doses (10(7)) with varying median time to death of ∼15 days, while at low doses (10(2)) all lobsters survived and no bacteria were present after 42 days. This work demonstrates the importance of the thickness of the A. viridans capsule to virulence in the American lobster.

Using Best Practices to Extract, Organize, and Reuse Embedded Decision Support Content Knowledge Rules from Mature Clinical Systems.

Clinical decision support (CDS) knowledge, embedded over time in mature medical systems, presents an interesting and complex opportunity for information organization, maintenance, and reuse. To have a holistic view of all decision support requires an in-depth understanding of each clinical system as well as expert knowledge of the latest evidence. This approach to clinical decision support presents an opportunity to unify and externalize the knowledge within rules-based decision support. Driven by an institutional need to prioritize decision support content for migration to new clinical systems, the Center for Knowledge Management and Health Information Technology teams applied their unique expertise to extract content from individual systems, organize it through a single extensible schema, and present it for discovery and reuse through a newly created Clinical Support Knowledge Acquisition and Archival Tool (CS-KAAT). CS-KAAT can build and maintain the underlying knowledge infrastructure needed by clinical systems.

Comparative tissue expression of American lobster (Homarus americanus) immune genes during bacterial and scuticociliate challenge.

The American lobster (Homarus americanus) fishery is the most economically significant fishery in Canada; although comparatively little is known about the lobsters' response to pathogenic challenge. This is the first study to investigate the expression of immune genes in tissues outside of the lobster hepatopancreas in response to challenges by the Gram-positive bacteria, Aerococcus viridans var. homari or the scuticociliate parasite, Anophryoides haemophila. The hepatopancreas has been regarded as the major humoral immune organ in crustaceans, but the contribution of other organs and tissues to the molecular immune response has largely been overlooked. This study used RT-qPCR to monitor the gene expression of several immune genes including three anti-lipopolysaccharide isoforms (ALF) Homame ALF-B1, Homame ALF-C1 and ALFHa-1, acute phase serum amyloid protein A (SAA), as well as thioredoxin and hexokinase, in antennal gland and gill tissues. Our findings indicate that the gene expression of the SAA and all ALF isoforms in the antennal gland and gill tissues increased in response to pathogenic challenge. However, there was differential expression of individual ALF isoforms that were dependent on both the tissue, and the pathogen used in the challenge. The gene expression changes of several immune genes were found to be higher in the antennal gland than have been previously reported for the hepatopancreas. This study demonstrates that increased immune gene expression from the gill and antennal gland over the course of pathogen induced disease contributes to the immune response of H. americanus.

Decreased Bioenergetic Health Index in monocytes isolated from the pericardial fluid and blood of post-operative cardiac surgery patients.

Monitoring the bioenergetics of leucocytes is now emerging as an important approach in translational research to detect mitochondrial dysfunction in blood or other patient samples. Using the mitochondrial stress test, which involves the sequential addition of mitochondrial inhibitors to adherent leucocytes, we have calculated a single value, the Bioenergetic Health Index (BHI), which represents the mitochondrial function in cells isolated from patients. In the present report, we assess the BHI of monocytes isolated from the post-operative blood and post-operative pericardial fluid (PO-PCF) from patients undergoing cardiac surgery. Analysis of the bioenergetics of monocytes isolated from patients' PO-PCF revealed a profound decrease in mitochondrial function compared with monocytes isolated from their blood or from healthy controls. Further, patient blood monocytes showed no significant difference in the individual energetic parameters from the mitochondrial stress test but, when integrated into the BHI evaluation, there was a significant decrease in BHI compared with healthy control monocytes. These data support the utility of BHI measurements in integrating the individual parameters from the mitochondrial stress test into a single value. Supporting our previous finding that the PO-PCF is pro-oxidant, we found that exposure of rat cardiomyocytes to PO-PCF caused a significant loss of mitochondrial membrane potential and increased reactive oxygen species (ROS). These findings support the hypothesis that integrated measures of bioenergetic health could have prognostic and diagnostic value in translational bioenergetics.

Antiinflammatory Activity of a Novel Folic Acid Targeted Conjugate of the mTOR Inhibitor Everolimus.

Folate receptor (FR)-ß has been identified as a promising target for antimacrophage and antiinflammatory therapies. In the present study, we investigated EC0565, a folic acid-derivative of everolimus, as a FR-specific inhibitor of the mammalian target of rapamycin (mTOR). Because of its amphiphilic nature, EC0565 was first evaluated for water solubility, critical micelle formation, stability in culture and FR-binding specificity. Using FR-expressing macrophages, the effect of EC0565 on mTOR signaling and cellular proliferation was studied. The pharmacokinetics, metabolism and bioavailability of EC0565 were studied in normal rats. The in vivo activity of EC0565 was assessed in rats with adjuvant arthritis, a "macrophage-rich" model with close resemblance to rheumatoid arthritis. EC0565 forms micellar aggregates in physiological buffers and demonstrates good water solubility as well as strong multivalent FR-binding capacity. EC0565 inhibited mTOR signaling in rat macrophages at nanomolar concentrations and induced G0/G1 cell cycle arrest in serum-starved RAW264.7 cells. Subcutaneously administered EC0565 in rats displayed good bioavailability and a relatively long half-life (~12 h). When given at 250 nmol/kg, EC0565 selectively inhibited proliferating cell nuclear antigen expression in thioglycollate-stimulated rat peritoneal cells. With limited dosing regimens, the antiarthritic activity of EC0565 was found superior to that of etanercept, everolimus and a nontargeted everolimus analog. The in vivo activity of EC0565 was also comparable to that of a folate-targeted aminopterin. Folate-targeted mTOR inhibition may be an effective way of suppressing activated macrophages in sites of inflammation, especially in nutrient-deprived conditions, such as in the arthritic joints. Further investigation and improvement upon the physical and biochemical properties of EC0565 are warranted.

Effects of dietary enrichment with a marine oil-based n-3 LCPUFA supplement in sows with predicted birth weight phenotypes on growth performance and carcass quality of offspring.

Effects of a marine oil-based n-3 LCPUFA supplement (mLCPUFA) fed from weaning until the end of the next lactation to sows with a predicted low litter birth weight (LBW) phenotype on growth performance and carcass quality of litters born to these sows were studied, based on the hypothesis that LBW litters would benefit most from mLCPUFA supplementation. Sows were allocated to be fed either standard corn/soybean meal-based gestation and lactation diets (CON), or the same diets enriched with 0.5% of the mLCPUFA supplement at the expense of corn. The growth performance from birth until slaughter of the litters with the lowest average birth weight in each treatment (n=24 per treatment) is reported in this paper. At weaning, each litter was split between two nursery pens with three to six pigs per pen. At the end of the 5-week nursery period, two barrows and two gilts from each litter that had individual birth weights closest to their litter average birth weight, were moved to experimental grow-finish pens (barn A), where they were housed as two pigs per pen, sorted by sex within litter. Remaining pigs in each litter were moved to another grow-finish barn (barn B) and kept in mixed-sex pens of up to 10 littermates. After 8 weeks, one of the two pigs in each pen in barn A was relocated to the pens holding their respective littermates in barn B. The remaining barrows and gilts were individually housed in the pens in barn A until slaughter. Maternal mLCPUFA supplementation increased docosahexaenoic acid (DHA) concentration in the brain, liver and Semitendinosus muscle of stillborn pigs (P<0.01), did not affect eicosapentaenoic acid and DHA concentrations in sow serum at the end of lactation, and did not affect average daily gain, average daily feed intake or feed utilization efficiency of the offspring. BW was higher (P<0.01) in the second half of the grow-finish phase in pigs from mLCPUFA sows compared with controls in barn A, where space and competition for feed was minimal, but not barn B. Carcass quality was not affected by treatment for pigs from barn A, but maternal mLCPUFA supplementation negatively affected carcass quality in pigs from barn B. Collectively, these results suggest that nutritional supplementation of sows can have lasting effects on litter development, but that feeding mLCPUFA to sows during gestation and lactation was not effective in improving growth rates or carcass quality of LBW litters.