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Obesity is associated with chronic inflammation and is a risk factor for insufficient milk production. Inflammation-mediated suppression of lipoprotein lipase could inhibit mammary uptake of long-chain fatty acids (LCFAs,16C).In an ancillary case control analysis, we investigated whether women with low milk production despite regular breast emptying have elevated inflammation and disrupted transfer of LCFAs from plasma into milk.Data and specimens from a low milk supply study and an exclusively breastfeeding control group were analyzed, with milk production measured by 24-hour test-weighing at 2-10 weeks postpartum. Low milk supply groups were defined as very low (VL,300 mL/d; n = 23) or moderate (MOD, ≥300 mL/d; n = 20) milk production, and compared to controls (≥699 mL/d; n = 18). Serum and milk fatty acids (weight% of total) were measured by gas chromatography, serum and milk TNF-α by ELISA, and serum high-sensitivity C-reactive protein (hsCRP) by clinical analyzer. Group differences were assessed by linear regression models, Chi-square exact tests, and Kruskal Wallis nonparametric tests.VL cases, as compared to MOD and controls, had higher prevalence of elevated serum hsCRP (5 mg/L, 57%, 15%, 22%, respectively, p = 0.004), detectable milk TNF-α (67%, 32%, 33%, respectively, p = 0.04), and obesity (78%, 40%, 22%, respectively, p = 0.003). VL cases had lower LCFAs in milk (Mean[±SD], 60[±3%]) compared to MOD (65[±4%]) and controls (66[±5%]; (p 0.001). Milk and serum LCFAs were strongly correlated in controls (r = 0.82, p 0.001), but not in MOD (r = 0.25, p = 0.30) or VL (r = 0.20, p = 0.41) groups (pint 0.001).Mothers with very low milk production have significantly higher obesity and inflammatory biomarkers, lower LFCAs in milk, and disrupted association between plasma and milk LCFAs. These data support the hypothesis that inflammation disrupts normal mammary gland fatty acid uptake. Further research should address impacts of inflammation and obesity on mammary fatty acid uptake for milk production.

Aims Free fatty acid receptor 4 (Ffar4) is a G-protein-coupled receptor for endogenous medium-/long-chain fatty acids that attenuates metabolic disease and inflammation. However, the function of Ffar4 in the heart is unclear. Given its putative beneficial role, we hypothesized that Ffar4 would protect the heart from pathologic stress. Methods and results In mice lacking Ffar4 (Ffar4KO), we found that Ffar4 is required for an adaptive response to pressure overload induced by transverse aortic constriction (TAC), identifying a novel cardioprotective function for Ffar4. Following TAC, remodelling was worsened in Ffar4KO hearts, with greater hypertrophy and contractile dysfunction. Transcriptome analysis 3-day post-TAC identified transcriptional deficits in genes associated with cytoplasmic phospholipase A2α signalling and oxylipin synthesis and the reduction of oxidative stress in Ffar4KO myocytes. In cultured adult cardiac myocytes, Ffar4 induced the production of the eicosapentaenoic acid (EPA)-derived, pro-resolving oxylipin 18-hydroxyeicosapentaenoic acid (18-HEPE). Furthermore, the activation of Ffar4 attenuated cardiac myocyte death from oxidative stress, while 18-HEPE rescued Ffar4KO myocytes. Systemically, Ffar4 maintained pro-resolving oxylipins and attenuated autoxidation basally, and increased pro-inflammatory and pro-resolving oxylipins, including 18-HEPE, in high-density lipoproteins post-TAC. In humans, Ffar4 expression decreased in heart failure, while the signalling-deficient Ffar4 R270H polymorphism correlated with eccentric remodelling in a large clinical cohort paralleling changes observed in Ffar4KO mice post-TAC. Conclusion Our data indicate that Ffar4 in cardiac myocytes responds to endogenous fatty acids, reducing oxidative injury, and protecting the heart from pathologic stress, with significant translational implications for targeting Ffar4 in cardiovascular disease.

In adipose, insulin functions to suppress intracellular lipolysis and secretion of nonesterified fatty acid (NEFA) into plasma. We applied glucose and NEFA minimal models (MM) following a frequently sampled intravenous glucose tolerance test (FSIVGTT) to assess glucose-specific and NEFA-specific insulin resistance. We used total NEFA and individual fatty acids in the NEFA MM, comparing the model parameters in metabolic syndrome (MetSyn) subjects ( n = 52) with optimally healthy controls (OptHC; n = 14). Results are reported as mean difference (95% confidence interval). Using the glucose MM, MetSyn subjects had lower [−73% (−82, −57)] sensitivity to insulin (Si) and higher [138% (44, 293)] acute insulin response to glucose (AIRg). Using the NEFA MM, MetSyn subjects had lower [−24% (−35, −13)] percent suppression, higher [32% (15, 52)] threshold glucose (gs), and a higher [81% (12, 192)] affinity constant altering NEFA secretion (ϕ). Comparing fatty acids, percent suppression was lower in myristic acid (MA) than in all other fatty acids, and the stearic acid (SA) response was so unique that it did not fit the NEFA MM. MA and SA percent of total were increased at 50 min after glucose injection, whereas oleic acid (OA) and palmitic acid (PA) were decreased ( P < 0.05). We conclude that the NEFA MM, as well as the response of individual NEFA fatty acids after a FSIVGTT, differ between OptHC and MetSyn subjects and that the NEFA MM parameters differ between individual fatty acids.

Background Regulation of the endoplasmic reticulum (ER) stress pathway is critical to mammary epithelial cell function throughout pregnancy, lactation, and involution. Treatment with trans-10, cis-12 conjugated linoleic acid (t10c12CLA) suppresses mammary lipogenesis and stimulates the ER stress pathway. The ER stress pathway includes tribbles pseudokinase 3 (TRB3), a protein that regulates cellular energy and insulin signaling. Objectives Our objective was to describe the effect of TRB3 deficiency on milk fat synthesis and determine if TRB3 deficiency protects against suppression of mammary lipogenesis. Methods First, mammary Trb3 expression was observed throughout pregnancy and lactation using ancillary microarray data (n = 4/time point). Second, intake, litter growth, and milk clot fatty acid (FA) profile of Trb3 knockout (KO) C57BL/6N mice were compared with wild-type (WT) and heterozygous (HET) mice throughout first (n ≥ 8/group) and second (n ≥ 6/group) lactation. Lastly, the interaction between Trb3 genotype and 2 treatments that suppress mammary lipogenesis, t10c12CLA and high safflower oil (HO) diet, was investigated in a 2 × 2 factorial design (n ≥ 6/group). Results Trb3 expression was higher during late pregnancy and lactation. Trb3 KO and HET mice had lower feed intake, dam weight, and litter growth throughout first, but not second, lactation than WT mice. Treatment with t10c12CLA decreased litter growth (28%; P, This study shows that, although the gene tribbles pseudokinase 3 (Trb3) is upregulated during lactation, it does not play an important role in regulating milk fat synthesis in lactating mice.

OBJECTIVES: Identification of physiologic factors that impede lactation is critical for developing interventions to support milk production. Our objective was to examine the role of tumor necrosis factor–alpha (TNF-A), a potent regulator of lipid metabolism, in the suppression of human lactation. We hypothesized that mothers with very low, versus sufficient milk production, would have elevated milk TNF-A and differential expression of TNF-A signaling pathways in the mammary epithelial cells (MECs). METHODS: Among mothers in a study of low milk supply, extracellular mammary epithelial cell mRNA was isolated from fresh milk fat and submitted to RNA-sequencing. Aligned and quantified reads were examined for differentially expressed genes using t-tests (DEG, p 0.05. Serum fasting triglyceride and BMI were significantly elevated in LMP versus HMP, P

Oxylipins are metabolites of polyunsaturated fatty acids that mediate cardiovascular health by attenuation of inflammation, vascular tone, hemostasis, and thrombosis. Very low‐density lipoproteins (VLDL) contain oxylipins, but it is unknown whether the liver regulates their concentrations. In this study, we used a perfused liver model to observe the effect of inflammatory lipopolysaccharide (LPS) challenge and soluble epoxide hydrolase inhibition (sEHi) on VLDL oxylipins. A compartmental model of deuterium‐labeled linoleic acid and palmitic acid incorporation into VLDL was also developed to assess the dependence of VLDL oxylipins on fatty acid incorporation rates. LPS decreased the total fatty acid VLDL content by 30% [6%,47%], and decreased final concentration of several oxylipins by a similar amount (13‐HOTrE, 35% [4%,55%], −1.3 nM; 9(10)‐EpODE, 29% [3%,49%], −2.0 nM; 15(16)‐EpODE, 29% [2%,49%], −1.6 nM; AA‐derived diols, 32% [5%,52%], −2.4 nM; 19(20)‐DiHDPA, 31% [7%,50%], −1.0 nM). However, the EPA‐derived epoxide, 17(18)‐EpETE, was decreased by 75% [49%,88%], (−0.52 nM) with LPS, double the suppression of other oxylipins. sEHi increased final concentration of DHA epoxide, 16(17)‐EpDPE, by 99% [35%,193%], (2.0 nM). Final VLDL‐oxylipin concentrations with LPS treatment were not correlated with linoleic acid kinetics, suggesting they were independently regulated under inflammatory conditions. We conclude that the liver regulates oxylipin incorporation into VLDL, and the oxylipin content is altered by LPS challenge and by inhibition of the epoxide hydrolase pathway. This provides evidence for delivery of systemic oxylipin signals by VLDL transport., Appearance of linoleate label in epoxides (EpOMEs) was greater than for alcohols (HODEs).

Background Methods to increase the amount of omega-3 (n-3) PUFAs in milk are desirable for neonatal health. The n-3 PUFA, α-linolenic acid (18:3n-3), can be elongated to EPA (20:5n-3) and DHA (22:6n-3). n-6 PUFAs suppress tissue n-3 PUFA incorporation, but the effect of SFAs is not clear. Objectives In this study, we compared the effects of SFAs and n-6 PUFAs on n-3 PUFA incorporation into milk and tissues of lactating mice and tissues of their offspring. Methods Female CD-1 mice were bred at 8 wk of age. All experimental diets included 3% flaxseed oil and were begun on day 8 of lactation: low-fat diet (LFD); high-SFA diet (SAT), with an additional 12% saturated oil; or high-linoleic-acid diet (HLA), with 12% high-linoleic-acid oil (% kcal, carbohydrates:fat:protein: LFD, 49:24:27; both SAT and HLA, 35:46:19; n = 5/treatment). After 5 d, pup stomach milk clot FA profiles, tissue FA profiles in dams and pups, and mammary and hepatic expression of lipid metabolism genes in dams were analyzed. Data were analyzed by ANOVA with treatment diet as a fixed effect. Results Dams in all groups had similar total milk fat concentrations, but both SAT and HLA decreased the concentration of n-3 PUFAs (SAT: -23%; HLA: -31%) compared with LFD, and HLA increased milk n-6 FAs by 347% compared with SAT. SAT pups had n-3 PUFA tissue concentrations similar to LFD, but HLA pups had lower n-3 PUFAs than SAT pups in multiple tissues (liver, -32%; kidney, -29%; heart, -28%; muscle, -18%). Mammary expression of lipid metabolism genes was mostly unchanged, but hepatic expression of elongases and desaturases was decreased with SAT compared with LFD [elongation of very-long-chain fatty acid (Elov)5, -42%; Elov6, -64%; fatty acid desaturase (Fads)1, -33%; Fads2, -44%]. Conclusions HLA decreased n-3 PUFA concentrations across multiple pup tissues compared with SAT. This suggests that high dietary n-6 PUFAs suppress n-3 PUFA incorporation in neonates.

OBJECTIVES: Oxylipins are derived from polyunsaturated fatty acids (PUFAs) and mediate inflammatory responses, but the time-dependent response of oxylipins to an acute inflammatory challenge in humans is unknown. Although omega-3 acid ethyl esters (O3EEs) reduce plasma triglycerides, it is unclear whether they alter the response of oxylipins in triglyceride rich lipoproteins (TGRLs). We investigated the effect of 3.4 g/d O3EEs on the oxylipin response to an inflammatory challenge. METHODS: Healthy young men (N = 16) received either placebo or 3.4 g/d O3EEs for 8–12 weeks in a crossover study design. After each treatment period, subjects underwent a low-dose endotoxin challenge (0.6 ng lipopolysaccharide (LPS)/kg body weight). Plasma samples were collected at baseline and 1, 2, 4, 8, 24, 48, 72, and 168 hours after LPS injection. TGRLs were separated from plasma, and esterified oxylipins were hydrolyzed and quantified by LC-MS/MS. Changes over time were assessed by the use of mixed models with repeated measures. Data is reported as mean (95% CI). RESULTS: The peak total oxylipin response occurred at 4 hours after LPS injection, with the exception of hydroxyeicosatetraenoic acids (HETEs). Regardless of treatment, the peak concentration of HETEs occurred at 2 hours. This timing differed from that of hydroxyoctadecadienoic acids (HODEs), which peaked at 4 hours (P = 0.004). Of the omega-3 derived oxylipins, O3EE treatment significantly increased hydroxyeicosapentaenoic acids (HEPEs) by 356% (133%, 792%), hydroxydocosahexaenoic acids (HDoHEs) by 172% (34%, 452%), and epoxydocosapentaeinoic acids (EpDPEs) by 316% (76%, 887%) compared to placebo at 4 hours after LPS challenge (P

OBJECTIVES: High fat meals can increase inflammation and plasma triglycerides. Poor suppression of postprandial adipocyte lipolysis results in higher levels of non-esterified fatty acids (NEFA) in insulin resistant subjects. This impaired suppression of NEFA may exacerbate hypertriglyceridemia following a high fat meal by increasing available fatty acids. We have shown myristic acid (MA) and stearic acid (SA) are less suppressed than other fatty acids 1 hour following a glucose challenge, and this pattern may indicate optimal adipocyte insulin sensitivity. It is unknown if the same pattern occurs in other contexts, such as high fat meal. METHODS: Plasma samples were collected from 12 obese male subjects at baseline and 4 time points following a high fat meal (1076 kcal, 39% kcal from saturated fat) on three visits. Meals contained either 0, 2, or 6 grams of a spice blend. Plasma was analyzed by GC-MS to measure multiple fatty acids, including MA, SA, palmitic acid (PA), oleic acid (OA), and linoleic acid (LA). RESULTS: At 1 hour following the high fat meal, MA and SA were less suppressed than OA, PA, and LA (P

BackgroundNon-­resolving activation of immune responses is central to the pathogenesis of heart failure (HF). Free fatty acid receptor 4 (Ffar4) is a G-protein coupled receptor (GPR) for medium-and long-chain fatty acids (FA) that regulates metabolism and attenuates inflammation in diabetes and obesity. Here, we tested the hypothesis that Ffar4 functions as a cardioprotective nutrient sensor that resolves inflammation to maintain cardiac homeostasis.MethodsMice with systemic deletion of Ffar4 (Ffar4KO) were subjected to pressure overload by transverse aortic constriction (TAC). Transcriptome analysis of cardiac myocytes was performed three days post-TAC. Additionally, Ffar4-mediated effects on inflammatory oxylipin production in cardiac myocytes and oxylipin composition in plasma lipoproteins were evaluated.ResultsIn Ffar4KO mice, TAC induced more severe remodeling, identifying an entirely novel cardioprotective role for Ffar4 in the heart. Transcriptome analysis 3-days post-TAC indicated a failure to induce cell death and inflammatory genes in Ffar4KO cardiac myocytes, as well as a specific failure to induce cytoplasmic phospholipase A2α (cPLA2α) signaling genes. In cardiac myocytes, Ffar4 signaling through cPLA2α-cytochrome p450 ω/ω-1 hydroxylase induced production of the EPA-derived anti-inflammatory oxylipin 18-hydroxyeicosapentaenoic acid (18-HEPE). Systemically, loss of Ffar4 altered oxylipin content in circulating plasma lipoproteins consistent with a loss of anti-inflammatory oxylipins at baseline, and inability to produce both pro-inflammatory and pro-resolving oxylipins following TAC. Finally, we confirmed that Ffar4 is expressed in human heart and down-regulated in HF.ConclusionsOur results identify a novel function for Ffar4 in the heart as a FA nutrient sensor that resolves inflammation to maintain cardiac homeostasis.

BACKGROUND Supplemental long-chain omega-3 (n-3) fatty acids (EPA and DHA) raise erythrocyte EPA + DHA [omega-3 index (O3I)] concentrations, but the magnitude or variability of this effect is unclear. OBJECTIVE The purpose of this study was to model the effects of supplemental EPA + DHA on the O3I. METHODS Deidentified data from 1422 individuals from 14 published n-3 intervention trials were included. Variables considered included dose, baseline O3I, sex, age, weight, height, chemical form [ethyl ester (EE) compared with triglyceride (TG)], and duration of treatment. The O3I was measured by the same method in all included studies. Variables were selected by stepwise regression using the Bayesian information criterion. RESULTS Individuals supplemented with EPA + DHA (n = 846) took a mean ± SD of 1983 ± 1297 mg/d, and the placebo controls (n = 576) took none. The mean duration of supplementation was 13.6 ± 6.0 wk. The O3I increased from 4.9% ± 1.7% to 8.1% ± 2.7% in the supplemented individuals ( P

Oxylipins are lipid mediators produced from polyunsaturated fatty acid (PUFA) metabolism, and are thought to be a molecular explanation for the diverse biological effects of PUFAs. Like PUFAs, oxylipins are distinguished by their omega-6 (n6) or omega-3 (n3) chemistry. We review the use of n6 oxylipins as biomarkers of disease and their use in diagnosis and risk assessment. We show cases where oxylipins derived from linoleate (LA) or arachidonate (AA) produced by the activities of lipoxygenase, cyclooxygenase, epoxygenase, ω/ω-1 hydroxylase, and autooxidation are useful as biomarkers or risk markers. HODEs, KODEs, EpOMEs, DiHOMEs, and other metabolites of LA as well as prostanoids, HETEs, KETEs, EpETrEs, and DiHETrEs, and other metabolites of AA were useful for understanding the different signaling environments in conditions from traumatic brain injury, to major coronary events, dyslipidemia, sepsis, and more. We next evaluate interventions that alter the concentrations of n6 oxylipins in plasma. We note the utility and response of each plasma fraction, and the generally increasing utility from the non-esterified, to the esterified, to the lipoprotein fractions. Finally, we review the effects which are specifically related to n6 oxylipins and most likely to be beneficial. Both n6 and n3 oxylipins work together in an exceedingly complex matrix to produce physiological effects. This overview should provide future investigators with important perspectives for the emerging utility of n6 oxylipins as products of n6 PUFAs in human health.

SummaryThe gut microbiota plays a key role in host metabolism. Toll-like receptor 5 (TLR5), a flagellin receptor, is required for gut microbiota homeostasis. Accordingly, TLR5-deficient (T5KO) mice are prone to develop microbiota-dependent metabolic syndrome. Here we observed that T5KO mice display elevated neutral lipids with a compositional increase of oleate [C18:1 (n9)] relative to wild-type littermates. Increased oleate contribution to hepatic lipids and liver SCD1 expression were both microbiota dependent. Analysis of short-chain fatty acids (SCFAs) and 13C-acetate label incorporation revealed elevated SCFA in ceca and hepatic portal blood and increased liver de novo lipogenesis in T5KO mice. Dietary SCFAs further aggravated metabolic syndrome in T5KO mice. Deletion of hepatic SCD1 not only prevented hepatic neutral lipid oleate enrichment but also ameliorated metabolic syndrome in T5KO mice. Collectively, these results underscore the key role of the gut microbiota-liver axis in the pathogenesis of metabolic diseases.

Objective: We have previously observed fatty acid epoxides, a class of potent anti-inflammatory oxylipins, in circulating VLDL. The source of these epoxides is unknown. Cytochrome P450 (CYP450) produces them via oxygenation of polyunsaturated fatty acids (PUFAs), and soluble epoxide hydrolase (sEH) converts them to diols. Our objectives were 1) to investigate if incorporation of epoxides into VLDL occurs via hepatic VLDL synthesis and 2) to determine if incorporation is modulated by inflammation or by inhibition of hepatic sEH. Approach and Results: A 2х2 factorial design was used for treatment assignment. Livers were isolated from rats treated with pro-inflammatory lipopolysaccharide (LPS, 10 mg/kg ip) or saline. AUDA, an inhibitor of sEH (10 μM), was included or excluded in the perfusate (Control, N=3; LPS, N=4; AUDA, N=4; LPS+AUDA, N=4). Livers were perfused for 180 minutes. VLDL was isolated by ultra-centrifugation, then analyzed by LC-MS/MS for oxylipin content. Analyzed epoxides and diols were derived from alpha-linolenic acid (ALA), linoleic acid (LA), arachidonic acid (AA), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA). Two-way ANOVA’s were used with triglyceride concentration as a covariate. Concentrations (nM) are reported as mean [95% CI]. DHA-derived epoxides increased with AUDA treatment (3.91 [3.01, 5.07]) compared to livers without AUDA (2.06 [1.58, 2.67]) (p=0.004), but other epoxides were unchanged by AUDA. EPA and ALA-derived epoxides decreased with LPS treatment (0.32 [0.22, 0.47]; 2.44 [2.07, 2.87]) compared to animals without LPS (0.73 [0.46, 1.16]; 3.28 [2.71, 3.96]) (p=0.01; 0.02). AA and DHA-derived diols decreased with LPS treatment (1.01 [0.82, 1.25]; 0.21 [0.17, 0.26]) compared to animals without LPS (1.46 [1.15, 1.86]; 0.31 [0.24, 0.39]) (p=0.03; 0.03). Conclusions: Treatment with LPS and AUDA have significant effects on incorporation of epoxides and diols into VLDL, supporting hepatic incorporation controlled by inflammation. Inflammation decreased select EPA- and ALA-derived epoxides. In contrast, sEH inhibition increased only DHA-derived epoxides. Surprisingly, in VLDL only epoxides derived from omega-3 fatty acids were affected by either inflammation or inhibition of sEH.

Background: Insulin resistance is a major contributor to metabolic syndrome, disrupting both glucose and non-esterified fatty acid (NEFA) dynamics through ineffective glucose clearance and decreased suppression of lipid droplet lipolysis. The minimal model of glucose dynamics is used for glycemic insulin sensitivity however it does not measure adipocyte insulin sensitivity, the primary determinant of plasma NEFA. An in-vivo approach to measuring adipocyte insulin sensitivity using NEFA is employed, comparing healthy and metabolic syndrome subjects. Both the models are employed to estimate insulin sensitivity and validate the NEFA approach. Objective: To test the use of NEFA kinetics to measure adipocyte insulin sensitivity compared to the glucose minimal model. Approach and results: Metabolic syndrome (n=56) and optimally healthy (n=14) subjects underwent a frequently sampled intravenous glucose tolerance test, and plasma analyzed for insulin, glucose, and NEFA. Insulin sensitivity ( S I ) and glucose effectiveness ( S G ) were calculated from the glucose minimal model. S I was 1.7 (mU/L) -1 min -1 and 0.40 (mU/L) -1 /min -1 and S G was 0.027 min -1 and 0.017 min -1 for the healthy and metabolic syndrome groups, respectively, indicating substantial glycemic insulin resistance in the latter. A model using glucose as the driver for NEFA kinetics was then applied. We found the initial rate of NEFA utilization by tissues (NU) was less, but the threshold glucose (tG) and glucose concentration required for a unit change in lipolysis inhibition ( G i ) were greater in metabolic syndrome verses healthy (NU: 0.050[0.045, 0.057] vs. 0.068[0.054, 0.086] p=0.03; tG: 6.7[6.2, 7.2] vs. 5.0[4.3, 5.9] p=0.001; G i : 0.30[0.25, 0.35] vs. 0.17[0.07, 0.27] p=0.02). No differences were found in initial rate of NEFA production or glucose utilization. Conclusion: Our results indicate that suppression of lipid-droplet lipolysis requires greater stimulus in metabolic syndrome compared to insulin sensitive adipocytes. Further, the rate of NEFA removal is less in metabolic syndrome. These results reveal components of insulin sensitivity not demonstrated by the glucose model. The NEFA model provides a measurement of adipocyte insulin sensitivity not captured by glycemic indices.

Military personnel engage in strenuous physical activity and load carriage. This study evaluated the role of body mass and of added mass on aerobic performance (uphill treadmill exercise) and pulmonary function. Performance on a traditional unloaded run test (4.8 km) was compared with performance on loaded tasks. Subjects performed an outdoor 4.8-km run and 4 maximal treadmill tests wearing loads of 0, 10, 20, and 30 kg. Subjects' pulmonary function (forced expired volume in 1 second [FEV1], forced vital capacity [FVC], and maximal voluntary ventilation [MVV]) was tested with each load, and peak values of heart rate, oxygen consumption ((Equation is included in full-text article.)), ventilation (VE), and respiratory exchange ratio (RER) were measured during each treadmill test. Performance on the 4.8-km run was correlated with treadmill performance, measured as time to exhaustion (TTE), with the strength of the correlation decreasing with load (r = 0.87 for 0 kg to 0.76 for 30 kg). Body mass was not correlated with TTE, other than among men with the 30-kg load (r = 0.48). During treadmill exercise, all peak responses other than RER decreased with load. Pulmonary function measures (FEV1, FVC, and MVV) decreased with load. Body mass was poorly correlated with treadmill performance, but added mass decreased performance. The decreased performance may be in part because of decreased pulmonary function. Unloaded 4.8-km run performance was correlated to unloaded uphill treadmill performance, but less so as load increased. Therefore, traditional run tests may not be an effective means of evaluating aerobic performance for military field operations.

This paper examines regulations for oil pipeline repair and considers some best practices to streamline permitting and assure compliance in the U.S. In the U.S., the federal government does not regulate construction or abandonment for oil pipelines, as it does with natural gas pipelines through the Federal Energy Regulatory Commission (the FERC). Instead, a number of federal, state, and local authorities have jurisdiction over oil pipelines and may regulate these activities differently. Because oil pipelines cross many jurisdictional boundaries, applicants find it challenging to navigate permitting requirements. For example, determining which permits are required for pipeline repair in U.S. waters can be difficult because, while such waters are defined by federal statute, the application of those rules is not uniform in all jurisdictions. In this paper, we present best practices for assuring compliance and streamlining the permitting process for oil pipeline construction, routine maintenance and repair in the U.S. To the extent possible, best practices mimic the FERC process, promote dialogue between regulators and oil pipeline companies, and foster a more transparent and effective environmental regulatory process. This paper presents the challenges encountered by the oil pipeline industry as a result of multiple agencies regulating the repair of oil pipelines. The paper then presents some discussion of the regulatory process and approaches to addressing challenges associated with this process. Following best practices to meet regulatory requirements is not only efficient and cost effective, but also helps assure long term regulatory compliance which affects the success of both small and larger scale projects.Copyright © 2010 by ASME

Background Myocardial infarction (MI) is a common cause of death, with approximately 175000 inpatient episodes of acute myocardial infarction in the UK in 2012. Following an MI the loss of cardiac myocytes triggers a remodelling process depositing extracellular matrix in the infarct region. Hypocontractile, damaged cardiac myocytes surround the infarct, forming a border zone. One of the key structural components regulating excitation-contraction coupling in the heart is the t-tubule network. Conventional confocal microscopy (resolution ˜100 nm) of isolated cardiac myocytes from the border zone post-MI has shown a loss of t-tubules. Here we apply, for the first time to our knowledge, serial block face scanning electron microscopy (SBF-SEM), to investigate the morphology of the t-tubule network within the infarct border zone cardiac myocytes to provide nano-scale structural details of the remodelling process. Methods A porcine model of MI was employed for this study. All animal work was approved by the University of Manchester local ethics committee and was covered by the necessary UK Home Office project and personal licences. Tissue (˜0.5 mm 3 ) was collected from the border and remote zones of the MI pigs and corresponding regions were also taken from control animals and processed for SBF-SEM. Blocks were imaged using an FEI Quanta 250 FEG SEM equipped with a Gatan 3View system. Serial images were collected at different magnifications ranging from 5.4 to 90.0 nm per pixel in the X-Y plane, while the cutting depth along the Z-axis was fixed at 50 nm for all the datasets. Images were segmented and rendered in Fiji or IMOD. Results 3D reconstruction of the t-tubule network from the left ventricle of control animals revealed a spoke-like arrangement, similar to that observed in human cardiac myocytes and other large mammals e.g. the sheep. Cardiac myocytes within the remote region of the infarcted heart have a t-tubule network indistinguishable from that of the control myocytes. In contrast, border zone myocytes showed large areas that were devoid of t-tubules. 3D modelling revealed that the surviving transverse tubules presented gross deformations, appearing to be the result of t-tubules fusing with each other to form a large complex that adopts a variety of orientations within the cardiac myocyte. Conclusion Employing SBF-SEM we have collected 3D datasets of cardiac myocytes in situ within the left ventricle of infarcted pigs at magnifications corresponding to ˜5 nm per pixel in the X-Y plane. This has allowed the resolution of nano-scale details of the remodelled t-tubules, including features such as the basal lamina. Together the loss of t-tubules within parts of the cell, coupled with the formation of super-tubule networks, provide novel structural insights towards unravelling the hypocontractile properties of the border zone cardiac myocytes.