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Facioscapulohumeral muscular dystrophy (FSHD) is an incurable skeletal myopathy. In absence of therapy, lifestyle factors impacting disease progression are important for clinical management. Monozygotic twins with FSHD often exhibit dramatically different disease progression, indicating existence of environmental disease modifiers. Here we analyse the USA National Registry for Myotonic Dystrophy & Facioscapulohumeral Dystrophy, comprising 511 FSHD1 patients followed up annually for an average of 8 years. This multimodal, longitudinal dataset comprises 189 baseline and 37 annually assessed features. We developed a workflow for prospective cohort analysis and identify cigarette smoking as associated with a two-fold increase in risk of facial and lower limb involvement in FSHD1 patients. Our definition of lower limb involvement includes inability to run and climb steps unaided, important functional outcomes for FSHD patients. We then employed an assay to test the effects of cigarette smoke extract on human myoblasts in vitro. Cigarette smoke extract drove disproportionate defects in proliferation and myogenic differentiation of FSHD1 patient-derived myoblasts, compared to matched controls. Mitochondrial function was also inordinately affected in FSHD1 myoblasts exposed to cigarette smoke extract, with increased mitochondrial membrane potential and mitochondrial radical oxygen species (mitoROS) generation. Our findings support recommending smoking cessation in clinical management of FSHD.

Asthma is a common and ubiquitous chronic respiratory disease that is associated with airway inflammation and hyperreactivity resulting in airway obstruction. It is now accepted that asthma is controlled by a combination of host genetics and environment in a rather complex fashion; however, the link between sensing of the environment and development and exacerbation of allergic lung inflammation is unclear. Human populations expressing cosegregating D299G and T399I polymorphisms in the TLR4 gene are associated with a decreased risk for asthma in adults along with hyporesponsiveness to inhaled LPS, the TLR4 ligand. However, these data do not account for other human genetic or environmental factors. Using a novel mouse strain that expresses homologous human TLR4 polymorphisms (TLR4-single nucleotide polymorphism [SNP]), we directly tested the effect of these TLR4 polymorphisms on in vivo responses to allergens using two models of induction. We report that intact TLR4 is required for allergic inflammation when using the OVA and LPS model of induction, as cellular and pathological benchmarks were diminished in both TLR4-SNP and TLR4-deficent mice. However, in the more clinically relevant model using house dust mite extract for induction, responses were enhanced in the TLR4-SNP mice, as evidenced by greater levels of eosinophilic inflammation, Th2 cytokine production, and house dust mite–specific IgG1 production compared with wild-type mice; however, mucus production and airway hyperreactivity were not affected. These results suggest that the TLR4 polymorphic variants (genes) interact differently with the allergic stimulation (environment).

Introduction There are no randomized controlled trials comparing low and high activated partial thromboplastin time (aPTT) targets in heparinized adult veno-arterial (VA) extracorporeal membrane oxygenation (ECMO) patients. Our systematic review and meta-analysis summarized complication rates in adult VA ECMO patients treated with low and high aPTT targets. Methods Studies published from January 2000 to May 2022 were identified using Pubmed, Embase, Cochrane Library, and LILACS (Latin American and Caribbean Health Sciences Literature). Studies were included if aPTT was primarily used to guide heparin anticoagulation. For the low aPTT group, we included studies where aPTT goal was ≤60 seconds and for the high aPTT group, we included studies where aPTT goal was ≥60 seconds. Proportional meta-analysis with a random effects model was used to calculate pooled complication rates for patients in the two aPTT groups. Results Twelve studies met inclusion criteria (5 in the low aPTT group and 7 in the high aPTT group). The pooled bleeding complication incidence for low aPTT studies was 53.6% (95% CI = 37.4%–69.4%, I2 = 60.8%) and for high aPTT studies was 43.8% (95% CI = 21.7%–67.1%, I2 = 91.8%). No studies in the low aPTT group reported overall thrombosis incidence, while three studies in the high aPTT group reported overall thrombosis incidence. The pooled thrombosis incidence for high aPTT studies was 16.1% (95% CI = 9.0%–24.5%, I2 = 13.1%). Conclusions Adult ECMO patients managed with low and high aPTT goals appeared to have similar bleeding and other complication rates further highlighting the need for a randomized controlled trial.

Supplemental Material for Outcomes of heparinized adult veno-arterial extracorporeal membrane oxygenation patients managed with low and high activated partial thromboplastin time targets: A systematic review and meta-analysis by Michael Mazzeffi, Dora Lin, Alberto Gonzalez-Almada, D Keegan Stombaugh, Jonathan Curley, Venkat Mangunta, Nicholas Teman, Leora T Yarboro and Robert Thiele in Perfusion.

Acute aortic dissection is a highly morbid condition with high mortality that requires emergent surgical evaluation and repair. The intraoperative management of acute aortic dissection requires the anesthesiologist to do far more than administer anesthesia and begins before the patient arrives at the operative theater. High-fidelity communication with the surgeon, knowledge of the surgical plan, knowledge of the anatomy of the dissection, and a nuanced understanding of aortic dissection pathophysiology are all critical aspects of anesthetic management.

Pulmonary hypertension (PH) is a devastating disease characterized by progressive elevation of pulmonary vascular resistance, right ventricular failure, and ultimately death. We have shown previously that insulin receptor substrate 2 (IRS2), a molecule highly critical to insulin resistance and metabolism, has an anti-inflammatory role in Th2-skewed lung inflammation and pulmonary vascular remodeling. Here, we investigated the hypothesis that IRS2 has an immunomodulatory role in human and experimental PH. Expression analysis showed that IRS2 was significantly decreased in the pulmonary vasculature of patients with pulmonary arterial hypertension and in rat models of PH. In mice, genetic ablation of IRS2 enhanced the hypoxia-induced signaling pathway of Akt and Forkhead box O1 (FOXO1) in the lung tissue and increased pulmonary vascular muscularization, proliferation, and perivascular macrophage recruitment. Furthermore, mice with homozygous IRS2 gene deletion showed a significant gene dosage-dependent increase in pulmonary vascular remodeling and right ventricular hypertrophy in response to hypoxia. Functional studies with bone marrow-derived macrophages isolated from homozygous IRS2 gene-deleted mice showed that hypoxia exposure led to enhancement of the Akt and ERK signaling pathway followed by increases in the pro-PH macrophage activation markers, vascular endothelial growth factor-A and arginase 1. Our data suggest that IRS2 contributes to anti-inflammatory effects by regulating macrophage activation and recruitment, which may limit the vascular inflammation, remodeling, and right ventricular hypertrophy that are seen in PH pathology. Restoring the IRS2 pathway may be an effective therapeutic approach for the treatment of PH and right heart failure.

Summary There are various health benefits associated with people consuming sufficient quantities of omega-3 fatty acids, with docosahexaenoic acid (DHA), in particular, impacting cardiovascular and mental health as well as influencing the immune response. Dietary deficiencies of omega-3 are common globally, and an emerging strategy to compensate is the enrichment of frequently consumed foods. Chicken meat represents a good target for enrichment as the fatty acid concentrations of the edible tissues can be easily altered by including omega-3–rich ingredients in their diets. However, the optimum duration of supplementation remains unclear. In this study, we describe the results of 2 studies in which the DHA-rich protist, Aurantiochytrium limacinum was fed to broilers at a rate of 0, 0.5, 1, and 2% of the diet. In the first study, the birds were supplemented for their whole lives (42 D), whereas in the second study, the birds were supplemented for the final 21-D fattening period only. Productivity parameters were affected in the whole-life study but were unaffected in the fattening-period study. In both studies, higher A. limacinum inclusion rates led to higher levels of DHA enrichment. Supplementation at a rate of 1% resulted in 49 and 97 mg DHA/100 g of breast or thigh meat, respectively, whereas 2% inclusion rate resulted in 97 and 156 mg DHA/100 g of breast or thigh meat, respectively, enough DHA to be considered a source of omega-3 in the European Union and Canada.

Summary Omega-3 fatty acids play an important role in various aspects of human health, but many people do not consume them in sufficient quantities, resulting in deficiencies in some populations. The enrichment of commonly consumed foods with omega-3 fatty acids has been proposed to address this deficit. Feeding omega-3–rich ingredients to animals can enrich their products, increasing the consumption by the population without requiring any major dietary changes. Eggs can be enriched to a high degree, and currently, omega-3–enriched eggs are widely available. Oftentimes, a missed opportunity for the poultry industry is in valorizing the spent hen. In this short-term study, a docosahexaenoic acid (DHA)–rich protist was fed to layer hens at three different inclusion levels to determine the degree of enrichment observed in the eggs and concomitantly in the breast, thigh, liver, and kidney. The addition of the protist ingredient had no negative impact on the bird health or performance. Significant increases in egg DHA concentration were observed, with 60, 164, 259, and 410 mg DHA/100g of egg collected from birds supplemented with the DHA-rich protist at a rate of 0, 0.5, 1, and 2.5% of the diet, respectively. This enrichment could increase the value of spent hen meat when used in the production of human and companion animal food as DHA-enriched products can be sold for a premium. Moreover, this enriched spent hen meat could potentially be used as a partial substitute for less-sustainable sources of dietary omega-3 including fish meal and fish oil.

Semaphorin 4A (Sema4A) exerts a stabilizing effect on human Treg cells in PBMC and CD4+ T cell cultures by engaging Plexin B1. Sema4A deficient mice display enhanced allergic airway inflammation accompanied by fewer Treg cells, while Sema4D deficient mice displayed reduced inflammation and increased Treg cell numbers even though both Sema4 subfamily members engage Plexin B1. The main objectives of this study were: 1. To compare the in vitro effects of Sema4A and Sema4D proteins on human Treg cells; and 2. To identify function-determining residues in Sema4A critical for binding to Plexin B1 based on Sema4D homology modeling. We report here that Sema4A and Sema4D display opposite effects on human Treg cells in in vitro PBMC cultures; Sema4D inhibited the CD4+CD25+Foxp3+ cell numbers and CD25/Foxp3 expression. Sema4A and Sema4D competitively bind to Plexin B1 in vitro and hence may be doing so in vivo as well. Bayesian Partitioning with Pattern Selection (BPPS) partitioned 4505 Sema domains from diverse organisms into subgroups based on distinguishing sequence patterns that are likely responsible for functional differences. BPPS groups Sema3 and Sema4 into one family and further separates Sema4A and Sema4D into distinct subfamilies. Residues distinctive of the Sema3,4 family and of Sema4A (and by homology of Sema4D) tend to cluster around the Plexin B1 binding site. This suggests that the residues both common to and distinctive of Sema4A and Sema4D may mediate binding to Plexin B1, with subfamily residues mediating functional specificity. We mutated the Sema4A-specific residues M198 and F223 to alanine; notably, F223 in Sema4A corresponds to alanine in Sema4D. Mutant proteins were assayed for Plexin B1-binding and Treg stimulation activities. The F223A mutant was unable to stimulate Treg stability in in vitro PBMC cultures despite binding Plexin B1 with an affinity similar to the WT protein. This research is a first step in generating potent mutant Sema4A molecules with stimulatory function for Treg cells with a view to designing immunotherapeutics for asthma.

All UK H&I laboratories and transplant units operate under a single national kidney offering policy, but there have been variations in approach regarding when to undertake the pre-transplant crossmatch test. In order to minimize cold ischaemia times for deceased donor kidney transplantation we sought to find ways to be able to report a crossmatch result as early as possible in the donation process. A panel of experts in transplant surgery, nephrology, specialist nursing in organ donation and H&I (all relevant UK laboratories represented) assessed evidence and opinion concerning five factors that relate to the effectiveness of the crossmatch process, as follows: when the result should be ready for reporting; what level of donor HLA typing is needed; crossmatch sample type and availability; fairness and equity; risks and patient safety. Guidelines aimed at improving practice based on these issues are presented, and we expect that following these will allow H&I laboratories to contribute to reducing CIT in deceased donor kidney transplantation.

Interleukin-4 (IL-4) is a four-α-helical bundle type I cytokine with broad pleiotropic actions on multiple lineages. Major actions of IL-4 were initially discovered for B and T cells, but this cytokine acts on more than a dozen different target cells spanning the innate and adaptive immune systems and is produced by multiple different cellular sources. While IL-4 was discovered just under 40 years ago in 1982, the interest in and discoveries related to this cytokine continue to markedly expand. There are important new advances related to its biological actions and to its mechanisms of signaling, including critical genes and downstream targets in a range of cell types. IL-4 is critical not only for careful control of immunoglobulin production but also related to inflammation, fibrosis, allergic reactions, and antitumor activity, with actions of IL-4 occurring through two different types of receptors, one of which is also used by IL-13, a closely related cytokine with partially overlapping actions. In this review, we cover critical older information but also highlight newer advances. An area of evolving interest relates to the therapeutic blockade of IL-4 signaling pathway to treat atopic dermatitis and asthma. Thus, this cytokine is historically important, and research in this area has both elucidated major biological pathways and led to therapeutic advances for diseases that affect millions of individuals.

SUMMARY Pelleting is commonly used in the production of poultry feed as it can reduce waste, kill pathogenic bacteria, and improve productivity. However, high temperatures used during production can negatively impact some feed ingredients. Omega-3 long-chain polyunsaturated fatty acid (n-3 LC-PUFA)-rich ingredients are often added to poultry feed to increase the n-3 LC-PUFA concentration of meat and eggs, adding value due to the health benefits associated with n-3 LC-PUFA consumption. Fish oils have traditionally been used for this purpose, but the n-3 LC-PUFA they contain are susceptible to oxidation, which can result in rancidity in enriched poultry products. Alternatively, protists that are primary producers of n-3 LC-PUFA are used. When provided in an unextracted form, these organisms could potentially protect the n-3 LC-PUFA they contain from oxidation. Aurantiochytrium limacinum is a marine protist, rich in docosahexaenoic acid (DHA), an n-3 LC-PUFA associated with various health benefits. We supplemented layer feed with A. limacinum and investigated the effect of pelleting on the recovery of DHA from feed and the efficiency of DHA transfer from feed to eggs. Pelleting did not decrease the concentration of DHA in feed. Also, no differences were observed between the groups fed mash or pelleted feeds in terms of the level of DHA enrichment of eggs. These results indicate that DHA, in the form of an A. limacinum biomass, is resistant to pelleting conditions and can be used as an ingredient in pelleted poultry diets to increase the DHA concentration of poultry products.

Stock-type mares (498 ± 9 kg BW; 12 ± 7 yr) were used in a completely randomized design for 56 d to test the hypothesis that concentrate fortification improves apparent digestion and enhances lean mass over the topline. Horses were stratified by age, BW, and BCS and randomly assigned to either a custom pelleted concentrate (CON; n = 13), or an iso-caloric, iso-nitrogenous pellet that included amino acid fortification, complexed trace minerals, and fermentation metabolites (FORT; n = 10). Concentrate was offered at a total 0.75% BW/d (as-fed) twice daily, and diets were designed to meet or exceed maintenance requirements for mature horses. Horses had ad libitum access to Coastal bermudagrass hay (7.4% CP, 67% NDF, and 40% ADF). Every 14 d BW and BCS were recorded, and ultrasound images were captured every 28 d. longissimus dorsi area (LDA) and subcutaneous fat thickness (FT) were measured between the 12th and 13th ribs (12th/13th) and 17th and 18th ribs (17th/18th). Intramuscular fat at the 17th/18th ribs and rump fat-thickness were also obtained. Horses were dosed with 10 g/d of titanium dioxide (TiO2) for 14 d to estimate forage dry matter intake (DMI). To account for diurnal variation, fecal samples were collected twice daily at 12-h intervals during the last 4 days, advancing by 3 h each day to represent a 24-h period. Fecal samples were composited by horse and analyzed for TiO2 to estimate fecal output and acid detergent insoluble ash was used to calculate forage DMI. To evaluate body composition, horses were infused with a 0.12 g/kg BW deuterium oxide (D2O) on d 0 and 56. Body fat percentage (BF) was determined by quantifying D2O in plasma samples collected at pre- and 4-h postinfusion via mass spectrometry. All data were analyzed using PROC MIXED (SAS v9.4). The model contained a fixed effect of diet; horse (diet) was a random effect. Horses receiving FORT gained 17th/18th FT (P < 0.01) and increased 17th/18th LDA from d 0 to 56 (P < 0.01) while 17th/18th FT and LDA were unchanged in CON. Regardless of diet, BF estimated by D2O infusion increased in all horses from d 0 to 56 (P < 0.01). Average hay DMI was 2.1% BW, but did not differ between diets. In this study, concentrate fortification did not significantly (P ≥ 0.27) affect apparent digestion. In conclusion, concentrate fortification may promote greater muscle development along the posterior topline.

1. The consumption of adequate amounts of the long-chain polyunsaturated omega-3 fatty acids (n-3 LC-PUFA) has been associated with beneficial effects on human health. Eggs are commonly con...

SUMMARY Changes in modern human diets have resulted in a deficiency in omega-3 fatty acids. Rectifying this deficiency and increasing dietary levels of omega-3 fatty acids, such as docosahexaenoic acid (DHA), can improve overall human health. Fish are a rich source of omega-3 fatty acids; however, low levels of consumption in many parts of the world, coupled with declining fish stocks, mean that alternative dietary omega-3 sources are required. The enrichment of commonly consumed foods has been suggested as a strategy that could increase omega-3 consumption. Eggs are an obvious target for enrichment as they are consumed worldwide and can have their omega-3 content altered through dietary manipulation of hens. In the current study, algae were included in the diet of hens, at 3 inclusion levels, in order to increase the omega-3 content of their eggs. For each treatment, the DHA content of eggs reached a plateau, proportional to the level of inclusion in the diet, which was then maintained for the duration of the experimental period. Higher inclusion levels reached this plateau quicker; however, the efficiency of transfer was reduced with increased dietary DHA. This maintenance of an increased egg DHA content over a prolonged period of time emphasizes the suitability of dietary supplementation with algae as a means to improve the nutritional value of hen's eggs.

1. The consumption of sufficient quantities of long chain omega-3 fatty acids (n-3 LCPUFA) from meat and other animal products can lead to a variety of health benefits in humans. The fatty acid content of poultry meat can be increased by feeding birds ingredients that are rich in n-3 LCFUFA 2. The effect of feeding a docosahexaenoic acid (DHA) rich Aurantiochytrium limacinum biomass (AURA) on the fatty acid content of breast and thigh tissues was investigated in a feeding trial with 2880 male Ross 308 broilers. The broiler diets were supplemented with either 0, 0.25, 0.5 or 1% AURA from day 21 to 42 of age. 3. Supplementation significantly increased the DHA content of both breast and thigh meat at an inclusion rate of 1% in the diet, leading to a total of 42 and 46 mg DHA/100 g of fresh breast or thigh tissue respectively. Significant increases in the tissue eicosapentaenoic acid (EPA) concentration were seen alongside a reduced omega-6/omega-3 ratio, improving the nutritional value of the meat for consumers and identifying supplementation of broiler diets with A. limacinum as an effective and sustainable method to increase n-3 LCPUFA consumption in the human population.

Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Health Service Executive Health and Wellbeing, Saolta University Healthcare Group OnBehalf Croí the West of Ireland Cardiac Foundation, Galway, Ireland and the National Institute for Prevention and Cardiovascular Health Introduction People living with obesity are at an increased risk of cardiovascular disease (CVD). While development of obesity is multifactorial, lifestyle modification is fundamental to obesity treatment and risk factor reduction. We sought to measure the effects of a structured lifestyle modification programme on the physical and mental health of people living with obesity. Purpose This study investigated the impact of a 10-week, community based, lifestyle modification programme on CVD risk factors in people living with obesity (BMI ≥35kg/m2 with a co-morbidity or BMI ≥40kg/m2) who were referred from a specialist bariatric service. Methods Delivered by an interdisciplinary team (Nurse, Dietitian & Physiotherapist) the programme included weekly group-based exercise sessions and health promotion workshops. A wide range of topics were addressed in workshops, including nutrition, food labels, emotional eating, physical activity, sedentary behaviour, stress management and CVD risk factor reduction. Outcomes were measured at initial and end of programme assessments. Results 1122 people participated in the intervention between 2013 and 2019 with 78% (n = 877) completing the programme. At initial assessment 26.7% of participants had a diagnosis of type 2 diabetes; 37.3% were at high or very high risk of CVD; 44.7% were hypertensive and 31.4% had a history of depression. Mean BMI was 47.0kg/m2 with 56.4% of participants having a BMI >45kg/m2. The intervention had significant positive impacts on key outcomes such as psychosocial health, lipid profiles, blood pressure, adiposity and cardiovascular fitness. One of the most significant outcomes observed was the improvement in psycho-social health. Scores of anxiety and depression, assessed using the HADS, decreased by 1.5 and 2.2 points respectively (p 3% of initial bodyweight. The percentage of participants achieving the recommended physical activity guidelines increased by 31% (p Conclusions A lifestyle modification programme delivered by an interdisciplinary team, aimed at individuals living with obesity, is not only acceptable to participants but also significantly reduces CVD risk factors. These findings should influence the design of future programmes and healthcare policies in Ireland and abroad.

Three experiments were conducted to evaluate the feasibility of using a commercial slow-release urea product (SRU, Optigen®, Alltech Inc., Nicholasville, KY, USA) as a partial replacement for vegetable protein sources in cattle diets. The first experiment was an in vitro rumen fermentation that evaluated the effect of replacing soybean meal (SBM) nitrogen with nitrogen from either SRU or free urea in diets varying in forage:concentrate ratios. The second experiment examined the effect of replacing SBM with SRU on in situ dry matter and nitrogen degradability in the rumen. In the third experiment, a feeding trial was conducted to evaluate the effect of replacing SBM (0% as-fed SRU) with 1% or 3% as-fed SRU on feed carbon footprint (CFP, total greenhouse gas emissions associated with the life cycle of feed raw materials) and the toxicity potential of SRU in growing beef cattle. Results showed that replacing SBM with SRU up to 1.3% did not negatively affect in vitro rumen fermentation parameters. Supplementing SRU favourably decreased ruminal accumulation of ammonia and lactic acid when compared to free urea. There was no significant effect on effective rumen degradability of dry matter and nitrogen when one-third of SBM was replaced by SRU in the in situ study. Compared with the 0% SRU diet, feed CFP decreased by 18% and 54% in 1% SRU and 3% SRU diets, respectively. Additionally, feeding up to 3% SRU diet to beef cattle did not affect health and intake, and blood hematological and biochemical indices were within the physiological range for healthy bulls, suggesting no indication of ammonia toxicity. Overall, these results indicate that SRU can be used as a sustainable alternative to partially replace vegetable protein sources in ruminant diets without compromising rumen function and health of ruminants.

INTRODUCTION This study evaluated a multimodal, simulation-based course in veterinary anesthesia integrated into an existing veterinary curriculum. METHODS A simulation-based, multimodal training course in clinical anesthesia was evaluated using outcomes from multiple levels of the Kirkpatrick Model of Training Evaluation. Cognitive and affective outcomes were evaluated before and after instruction. Head-mounted cameras were used to record clinical performance during students' first live patient anesthesia experience in the curriculum. RESULTS Pretest-posttest analysis of cognitive and affective outcomes for course participants revealed significant increases in knowledge and self-efficacy. Course participants received higher ratings on clinical task performance and professional skills (ie, communication and collaboration) compared with a matched control group when evaluated by blinded, external raters using a standardized rubric. CONCLUSIONS These findings indicate that implementing a multimodal anesthesia simulation-based course directly into the curriculum can enhance cognitive and affective outcomes and prepare students for subsequent anesthesia-related patient care experiences.

Psychological stress is known to have profound effects on immune function and to promote inflammatory conditions. Elevated circulating levels of cytokines associated with stress are known to increase the risk to several diseases, but little is known about this mechanism. This study assessed the role of T cells on cytokine levels after exposure to stress in the learned helplessness paradigm. Adoptive transfer of CD4+ T cells into Rag2-/- mice did not change cytokine levels to stress while CD8+ T cells resulted in an increase in TNF-α, IL-6 and IFN-γ in stressed Rag2-/- mice. Moreover, depletion of CD8+ T cells in WT mice abolished these cytokine responses to stress. Corticosterone and behavioral stress responsiveness was impaired in Rag2-/- mice reconstituted with CD8+ T cells. Notably, depletion of these cells in WT mice had no effect on behavior or corticosterone levels. Exposure to stress did not change the expression of canonical markers of T cell activation including CD62L and CD44 or modified intracellular cytokine content, suggesting that they are not the main producers of circulating cytokines in response to stress. These results show that CD8+ T cells promote TNF-α, IL-6 and IFN-γ responses to stress, possibly by stimulating non-lymphoid cells.

INTRODUCTION: Emerging evidence suggests that effective treatment of glioblastoma (GBM), the most common and deadly form of adult primary brain cancer, will likely require concurrent treatment of multiple aspects of tumor pathobiology to overcome tumor heterogeneity and the complex tumor-supporting microenvironment. Recent studies in non-central nervous system (CNS) tumor cells have demonstrated that oxaliplatin (OXA) can induce multi-faceted anti-tumor effects, in particular at drug concentrations below those required to induce apoptosis. These findings motivated re-investigation of OXA for the treatment of GBM. METHODS: The effects of OXA on murine KR158 and GL261 glioma cells including cell growth, cell death, inhibition of signal transducer and activator of transcription (STAT) activity, O-6-methylguanine-DNA methyltransferase (MGMT) expression, and immunogenic cell death (ICD) initiation, were evaluated by cytotoxicity assays, Western blot analysis, STAT3-luciferase reporter assays, qRT-PCR assays, and flow cytometry. Chemical inhibitors of endoplasmic reticulum (ER) stress were used to investigate the contribution of this cell damage response to the observed OXA effects. The effect of OXA on bone marrowderived macrophages (BMDM) exposed to glioma conditioned media (GCM) was also analyzed by Western blot analysis. RESULTS: We identified the OXA concentration threshold for induction of apoptosis and from this determined the drug dose and treatment period for sub-cytotoxic treatments of glioma cells. Under these experimental conditions, OXA reduced STAT3 activity, reduced MGMT levels and increased temozolomide sensitivity. In addition, there was evidence of immunogenic cell death (elevated EIF2α phosphorylation and calreticulin exposure) following prolonged OXA treatment. Notably, inhibition of ER stress reversed the OXA-mediated inhibition of STAT3 activity and MGMT expression in the tumor cells. In BMDMs exposed to GCM, OXA also reduced levels of phosphorylated STAT3 and decreased expression of Arginase 1, an enzyme known to contribute to pro-tumor functions in the tumor-immune environment. CONCLUSIONS: OXA can induce notable multi-faceted biological effects in glioma cells and BMDMs at relatively low drug concentrations. These findings may have significant therapeutic relevance against GBM and warrant further investigation.

Targeted selective treatment (TST) has been proposed as a sustainable method of gastrointestinal nematode control that reduces the number of anthelmintic treatments administered, thereby preserving a susceptible nematode population in refugia. In order to minimise the impact of withholding treatment on animal performance, animals that would benefit most should be selected for treatment. However, the most suitable criteria for selecting which animals to treat remain a subject of research. The impact of implementing a TST strategy based on lamb live weight was investigated, and whether heavy lambs were more resilient than light lambs if left untreated. The study was conducted using weaned lambs on 3 sheep farms, and over 2 years. On each farm lambs were weighed and divided into heavy, medium and light weight classes. Within the heavy (n = 225) and light (n = 218) weight classes lambs were randomly allocated to two treatments; anthelmintic treatment or no anthelmintic treatment. All lambs in the medium weight class were treated. Animal performance and parasitological parameters were assessed over a 28-day period. Anthelmintic treatment had a significant effect (P 0.01) on faecal egg count, average daily weight gain, body condition score and dag score; there was no treatment by weight class interaction. Anthelmintic treatment had no effect on plasma pepsinogen concentration or on blood leukocyte numbers. Withholding anthelmintic treatment from lambs therefore had a negative effect on worm egg count and animal performance with no evidence that heavy lambs were more resilient than light lambs when left untreated.

Docosahexaenoic acid (DHA), is an omega 3 fatty acid (n-3 FA) that has been shown to play a role in canine growth and physiological integrity and improvements in skin and coat condition. However, potential adverse effects of n-3 FA specifically, impaired cellular immunity has been observed in dogs fed diets with elevated levels of n-3 FA. As such, a safe upper limit (SUL) for total n-3 FAs (DHA and EPA) in dogs has been established. Considering this SUL, sensitive methods detecting DHA in blood serum as a biomarker when conducting n-3 FA supplementation trials involving dogs are required. In this study, an LC-ESI-MS/MS method of DHA detection in dog serum was validated and verified. Recovery of DHA was optimized and parallelism tests were conducted with spiked samples demonstrating that the serum matrix did not interfere with quantitation. The stability of DHA in serum was also investigated, with −80 °C considered suitable when storing samples for up to six months. The method was linear over a calibration range of 1–500 μg/mL and precision and accuracy were found to meet the requirements for validation. This method was verified in an alternative laboratory using a different analytical system and operator, with the results meeting the criteria for verification.

OBJECTIVE The aim of this experiment was to evaluate the effect of dietary supplementation with the docosahexaenoic acid (DHA)-rich microalgae, Aurantiochytrium limacinum (AURA) on pig performance, carcass traits, and the fatty acid composition of pork Longissimus lumborum (LL) and backfat. METHODS A total of 144 Pig Improvement Company (PIC)×Goland finishing pigs (72 females and 72 castrated males) of mean weight 117.1 (±13.1) kg were blocked by sex and body weight and provided with 0% or 1% AURA in isonutritive and isocaloric diets. A total of 24 pens provided 12 replicates per treatment. Animals were weighed on day 0 and 28 with feed and water intake recorded per pen. After 31 days supplementation (28 days of study and 3 days until the slaughtering date) three animals per pen (n = 72) were slaughtered and the LL and backfat thickness, lean meat content and dressing percentage were recorded for the carcasses. The fatty acid (FA) profile of the LL and backfat was established by direct FA methyl ester synthesis. RESULTS No differences were observed for any performance parameters or carcass traits. Supplementation with AURA resulted in significant changes to the FA profiles of both the LL and backfat with male and female pigs responding differently to supplementation in terms of particular FAs. Overall, pork LL samples had significantly higher eicosapentaenoic acid (p