Maximal Strength, Muscle Activation, and Bar Velocity Comparisons Between Squatting With a Traditional or Safety Squat Bar.

ABSTRACT: Vantrease, WC, Townsend, JR, Sapp, PA, Henry, RN, and Johnson, KD. Maximal strength, muscle activation, and bar velocity comparisons between squatting with a traditional or safety squat bar. J Strength Cond Res 35(2S): S1-S5, 2021-The purpose of this study was to compare strength, muscle activation, and bar velocity between the traditional (TRAD) and safety squat bar (SSB) back squat. Thirty-two men (21.94 ± 3.1 years, 1.78 ± 0.8 m, 81.7 ± 10.1 kg) volunteered to complete this randomized, crossover-design study. Subjects completed 2 separate 1 repetition maximum (1RM) sessions using either the TRAD or SSB. Subsequently, subjects completed 1 session of 3 repetitions at 65 and 85% of their 1RM for each squat condition (SSB & TRAD). Peak muscle activation of 7 muscles from the lower body and trunk was recorded through surface electromyography (EMG), and mean velocity (MV) was recorded by a linear transducer. Electromyography and MV were analyzed by a 2 × 2 (bar × load) repeated-measures analysis of variance. A Pearson correlation was used to determine the relationship of 1RM load between bars. Squat 1RM was significantly higher (p < 0.001; 11.6%) for TRAD (144.7 kg) compared with SSB (128.8 kg), and a strong correlation (r = 0.94) was observed between 1RM values of each bar. A significant main effect was seen in EMG (p < 0.001) and MV for load (p < 0.001). No significant bar × load interaction was observed between conditions for any EMG or bar velocity measure (p > 0.05). The SSB produces similar muscle activation and bar velocities compared with the TRAD at relative intensities. However, absolute loads should be adjusted when changing squat bars during a training cycle.

The association of serum interleukin-6 levels with clinical outcomes in antineutrophil cytoplasmic antibody-associated vasculitis.

OBJECTIVE: To investigate serum IL-6 (sIL-6) levels during active disease, complete remission (CR), and relapse in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV), and to explore the association of changes in sIL-6 with clinical outcomes. METHODS: sIL-6 levels were measured at baseline and longitudinally over 18 months, in 78 patients with AAV enrolled in a randomized controlled trial comparing treatment with either rituximab (RTX) or cyclophosphamide (CYC)/azathioprine (AZA). Outcome variables included baseline clinical features, ANCA specificity, disease activity (active disease versus CR), time to relapse events, B cell repopulation, and ANCA titer increases. RESULTS: At baseline, sIL6 levels were detectable in 81% of patients; 73% (n = 57) of subjects were proteinase 3 (PR3)-ANCA positive, sIL-6 levels were higher in subjects with PR3-ANCAs and positively correlated with their levels (rs = 0.36,p < 0.01), but not with levels of myeloperoxidase (MPO)-ANCA (rs = -0.17,p = 0.47). Higher baseline sIL-6 levels were associated with PR3-ANCA positivity, fever, pulmonary nodules/cavities, conductive deafness, and absence of urinary red blood cell casts (p < 0.05). Baseline sIL6 levels did not predict CR at month 6 (p = 0.71), and the median sIL-6 level declined from baseline with induction therapy, regardless of CR achievement. An increase in sIL-6 during CR was a predictor for subsequent severe relapse in RTX-treated patients (hazard ratio (HR):7.24,p = 0.01), but not in CYC/AZA-treated patients (HR:0.62,p = 0.50). In contrast, a sIL-6 increase did not predict B cell repopulation or ANCA titer increase in either treatment arm (p > 0.05). CONCLUSION: At baseline, sIL-6 concentrations correlate with PR3-ANCA titers and are associated with specific clinical manifestations of AAV. Baseline sIL6 concentrations do not predict CR at 6 months, but the increase in sIL-6 concentrations during CR is associated with subsequent severe relapse among RTX-treated patients. Further investigation into the mechanistic role of IL6 in AAV might lead to identifying this pathway as a potential therapeutic target in this disease.

On-Board Sensor-Based NO x Emissions from Heavy-Duty Diesel Vehicles.

Real-world nitrogen oxides (NO x) emissions were estimated using on-board sensor readings from 72 heavy-duty diesel vehicles (HDDVs) equipped with a Selective Catalytic Reduction (SCR) system in California. The results showed that there were large differences between in-use and certification NO x emissions, with 12 HDDVs emitting more than three times the standard during hot-running and idling operations in the real world. The overall NO x conversion efficiencies of the SCR system on many vehicles were well below the 90% threshold that is expected for an efficient SCR system, even when the SCR system was above the optimum operating temperature threshold of 250 °C. This could potentially be associated with SCR catalyst deterioration on some engines. The Not-to-Exceed (NTE) requirements currently used by the heavy-duty in-use compliance program were evaluated using on-board NO x sensor data. Valid NTE events covered only 4.2-16.4% of the engine operation and 6.6-34.6% of the estimated NO x emissions. This work shows that low cost on-board NO x sensors are a convenient tool to monitor in-use NO x emissions in real-time, evaluate the SCR system performance, and identify vehicle operating modes with high NO x emissions. This information can inform certification and compliance programs to ensure low in-use NO x emissions.

Gasoline Particulate Filters as an Effective Tool to Reduce Particulate and Polycyclic Aromatic Hydrocarbon Emissions from Gasoline Direct Injection (GDI) Vehicles: A Case Study with Two GDI Vehicles.

We assessed the gaseous, particulate, and genotoxic pollutants from two current technology gasoline direct injection vehicles when tested in their original configuration and with a catalyzed gasoline particulate filter (GPF). Testing was conducted over the LA92 and US06 Supplemental Federal Test Procedure (US06) driving cycles on typical California E10 fuel. The use of a GPF did not show any fuel economy and carbon dioxide (CO2) emission penalties, while the emissions of total hydrocarbons (THC), carbon monoxide (CO), and nitrogen oxides (NOx) were generally reduced. Our results showed dramatic reductions in particulate matter (PM) mass, black carbon, and total and solid particle number emissions with the use of GPFs for both vehicles over the LA92 and US06 cycles. Particle size distributions were primarily bimodal in nature, with accumulation mode particles dominating the distribution profile and their concentrations being higher during the cold-start period of the cycle. Polycyclic aromatic hydrocarbons (PAHs) and nitrated PAHs were quantified in both the vapor and particle phases of the PM, with the GPF-equipped vehicles practically eliminating most of these species in the exhaust. For the stock vehicles, 2-3 ring compounds and heavier 5-6 ring compounds were observed in the PM, whereas the vapor phase was dominated mostly by 2-3 ring aromatic compounds.

Detailed Analysis of Criteria and Particle Emissions from a Very Large Crude Carrier Using a Novel ECA Fuel.

Ocean going vessels (OGVs) operating within emission control areas (ECA) are required to use fuels with ≤0.1 wt % sulfur. Up to now only distillate fuels could meet the sulfur limits. Recently refiners created a novel low-sulfur heavy-fuel oil (LSHFO) meeting the sulfur limits so questions were posed whether nitric oxide (NOx) and particulate matter (PM) emissions were the same for the two fuels. This project characterized criteria pollutants and undertook a detailed analysis of PM emissions from a very large crude oil carrier (VLCC) using a distillate ECA fuel (MGO) and novel LSHFO. Results showed emission factors of NOx were ∼5% higher with MGO than LSHFO. PM2.5 emission factors were ∼3 times higher with LSHFO than MGO, while both were below values reported by Lloyds, U.S. EPA and CARB. A detailed analysis of PM revealed it was >90% organic carbon (OC) for both fuels. Elemental carbon (EC) and soot measured with an AVL microsoot sensor (MSS) reflected black carbon. PM size distributions showed unimodal peaks for both MGO (20-30 nm) and LSHFO (30-50 nm). Particle number (PN) emissions were 28% and 17% higher with the PPS-M compared to the SMPS for LSHFO and MGO, respectively.

T cell PKCδ kinase inactivation induces lupus-like autoimmunity in mice.

Genetic and environmental factors contribute to the onset and progression of lupus. CD4+ T cells from patients with active lupus show a decreased ERK signaling pathway, which causes changes in gene expression. The defect points to its upstream regulator, PKCδ, which exhibits a deficient activity due to oxidative stress. Our aim was to investigate the effect of a defective PKCδ in the development of lupus. We generated a double transgenic C57BL6 × SJL mouse that expresses a doxycycline-induced dominant negative PKCδ (dnPKCδ) in T cells. The transgenic mice displayed decreased T cell ERK signaling, decreased DNMT1 expression and overexpression of methylation sensitive genes involved in the exaggerated immune response in the pathogenesis of lupus. The mice developed anti-dsDNA autoantibodies and glomerulonephritis with IgG deposition. The study indicates common pathogenic mechanisms with human lupus, suggesting that environmentally-mediated T cell PKCδ inactivation plays a causative role in lupus.

PF-1355, a mechanism-based myeloperoxidase inhibitor, prevents immune complex vasculitis and anti-glomerular basement membrane glomerulonephritis.

Small vessel vasculitis is a life-threatening condition and patients typically present with renal and pulmonary injury. Disease pathogenesis is associated with neutrophil accumulation, activation, and oxidative damage, the latter being driven in large part by myeloperoxidase (MPO), which generates hypochlorous acid among other oxidants. MPO has been associated with vasculitis, disseminated vascular inflammation typically involving pulmonary and renal microvasculature and often resulting in critical consequences. MPO contributes to vascular injury by 1) catabolizing nitric oxide, impairing vasomotor function; 2) causing oxidative damage to lipoproteins and endothelial cells, leading to atherosclerosis; and 3) stimulating formation of neutrophil extracellular traps, resulting in vessel occlusion and thrombosis. Here we report a selective 2-thiouracil mechanism-based MPO inhibitor (PF-1355 [2-(6-(2,5-dimethoxyphenyl)-4-oxo-2-thioxo-3,4-dihydropyrimidin-1(2H)-yl)acetamide) and demonstrate that MPO is a critical mediator of vasculitis in mouse disease models. A pharmacokinetic/pharmacodynamic response model of PF-1355 exposure in relation with MPO activity was derived from mouse peritonitis. The contribution of MPO activity to vasculitis was then examined in an immune complex model of pulmonary disease. Oral administration of PF-1355 reduced plasma MPO activity, vascular edema, neutrophil recruitment, and elevated circulating cytokines. In a model of anti-glomerular basement membrane disease, formerly known as Goodpasture disease, albuminuria and chronic renal dysfunction were completely suppressed by PF-1355 treatment. This study shows that MPO activity is critical in driving immune complex vasculitis and provides confidence in testing the hypothesis that MPO inhibition will provide benefit in treating human vasculitic diseases.

CD4(+) T cells epigenetically modified by oxidative stress cause lupus-like autoimmunity in mice.

Lupus develops when genetically predisposed people encounter environmental agents such as UV light, silica, infections and cigarette smoke that cause oxidative stress, but how oxidative damage modifies the immune system to cause lupus flares is unknown. We previously showed that oxidizing agents decreased ERK pathway signaling in human T cells, decreased DNA methyltransferase 1 and caused demethylation and overexpression of genes similar to those from patients with active lupus. The current study tested whether oxidant-treated T cells can induce lupus in mice. We adoptively transferred CD4(+) T cells treated in vitro with oxidants hydrogen peroxide or nitric oxide or the demethylating agent 5-azacytidine into syngeneic mice and studied the development and severity of lupus in the recipients. Disease severity was assessed by measuring anti-dsDNA antibodies, proteinuria, hematuria and by histopathology of kidney tissues. The effect of the oxidants on expression of CD40L, CD70, KirL1 and DNMT1 genes and CD40L protein in the treated CD4(+) T cells was assessed by Q-RT-PCR and flow cytometry. H2O2 and ONOO(-) decreased Dnmt1 expression in CD4(+) T cells and caused the upregulation of genes known to be suppressed by DNA methylation in patients with lupus and animal models of SLE. Adoptive transfer of oxidant-treated CD4(+) T cells into syngeneic recipients resulted in the induction of anti-dsDNA antibody and glomerulonephritis. The results show that oxidative stress may contribute to lupus disease by inhibiting ERK pathway signaling in T cells leading to DNA demethylation, upregulation of immune genes and autoreactivity.

Observer error, dental wear, and the inference of new world sundadonty.

Dental morphology provides important information on human evolution and interpopulation relationships. Dental wear is one of the major limitations of morphological data analysis. Wear figures heavily in existing debates about patterns of New World dental variation with some scholars finding evidence for a more generalized dentition in early New World populations (Powell: Doctoral Dissertation, Texas A&M University, TX (1995)) and others questioning these findings based on the probable effects of dental wear on trait scores (Turner, The First Americans: the Pleistocene Colonization of the New World. San Francisco: California Academy of Sciences (2002) 123-158; Turner: Am J Phys Anthropol 130 (2006) 455-461; Turner and Scott, Handbook of paleoanthropology, Vol. III: Phylogeny of Hominids. New York: Springer (2007) 1901-1941). Here we evaluate these competing claims using data from the Early Archaic Windover sample. Results confirm the dental distinctiveness of Windover with respect to other Old World Asian (i.e., sinodont/sundadont) populations. However, comparison of our results to those of Powell (1995) also highlights significant interobserver error. Statistical analysis of matched wear and morphology scores suggests trait downgrading for some traits. Patterns of missing data present a more challenging (and potentially serious) problem. Use of Little's MCAR test for missing data mechanisms indicates a complex process of data collection in which incidental and opportunistic recording of both highly worn and unerupted teeth introduce a "missing not at random" mechanism into our dataset that biases dental trait frequencies. We conclude that patterns of missingness and formal research designs for "planned missingness" are needed to help mitigate this bias.

Optimization of vancomycin dosing in very low-birth-weight preterm neonates.

OBJECTIVE: To compare vancomycin serum trough concentrations and 24-hour area under the serum concentration-versus-time curve (AUC24) among very low-birth-weight (VLBW) premature infants before and after implementation of an institution-wide increase in neonatal vancomycin dosing. STUDY DESIGN: We performed a retrospective analysis of vancomycin concentrations among preterm VLBW neonates before (2007-2010) and after (2010-2013) implementation of a new vancomycin dosing protocol consisting of increased vancomycin daily dose and frequency of administration. RESULTS: Neonates weighing < 1,500 g and receiving the new vancomycin dosing regimen had lower rates of undetectable trough concentrations (24 vs. 50%, p = 0.04), higher median trough concentrations (10.8 vs. 5.9 µg/mL, p = 0.003), a higher proportion of goal trough concentrations of 10 to 20 µg/mL (35 vs. 4%, p = 0.005), and a significantly higher vancomycin AUC24 (438 vs. 320 mg·h/L, p = 0.004) compared with historical controls. CONCLUSION: Increasing the vancomycin daily dose and dosing frequency led to an increase in vancomycin trough concentrations and AUC24, and a decrease in the proportion of undetectable (< 5.0 µg/mL) troughs, without an increase in toxicity among VLBW premature neonates.

Assessing the impacts of ethanol and isobutanol on gaseous and particulate emissions from flexible fuel vehicles.

This study investigated the effects of higher ethanol blends and an isobutanol blend on the criteria emissions, fuel economy, gaseous toxic pollutants, and particulate emissions from two flexible-fuel vehicles equipped with spark ignition engines, with one wall-guided direct injection and one port fuel injection configuration. Both vehicles were tested over triplicate Federal Test Procedure (FTP) and Unified Cycles (UC) using a chassis dynamometer. Emissions of nonmethane hydrocarbons (NMHC) and carbon monoxide (CO) showed some statistically significant reductions with higher alcohol fuels, while total hydrocarbons (THC) and nitrogen oxides (NOx) did not show strong fuel effects. Acetaldehyde emissions exhibited sharp increases with higher ethanol blends for both vehicles, whereas butyraldehyde emissions showed higher emissions for the butanol blend relative to the ethanol blends at a statistically significant level. Particulate matter (PM) mass, number, and soot mass emissions showed strong reductions with increasing alcohol content in gasoline. Particulate emissions were found to be clearly influenced by certain fuel parameters including oxygen content, hydrogen content, and aromatics content.

Comparison of particle mass and solid particle number (SPN) emissions from a heavy-duty diesel vehicle under on-road driving conditions and a standard testing cycle.

It is important to understand the differences between emissions from standard laboratory testing cycles and those from actual on-road driving conditions, especially for solid particle number (SPN) emissions now being regulated in Europe. This study compared particle mass and SPN emissions from a heavy-duty diesel vehicle operating over the urban dynamometer driving schedule (UDDS) and actual on-road driving conditions. Particle mass emissions were calculated using the integrated particle size distribution (IPSD) method and called MIPSD. The MIPSD emissions for the UDDS and on-road tests were more than 6 times lower than the U.S. 2007 heavy-duty particulate matter (PM) mass standard. The MIPSD emissions for the UDDS fell between those for the on-road uphill and downhill driving. SPN and MIPSD measurements were dominated by nucleation particles for the UDDS and uphill driving and by accumulation mode particles for cruise and downhill driving. The SPN emissions were ∼ 3 times lower than the Euro 6 heavy-duty SPN limit for the UDDS and downhill driving and ∼ 4-5 times higher than the Euro 6 SPN limit for the more aggressive uphill driving; however, it is likely that most of the "solid" particles measured under these conditions were associated with a combination release of stored sulfates and enhanced sulfate formation associated with high exhaust temperatures, leading to growth of volatile particles into the solid particle counting range above 23 nm. Except for these conditions, a linear relationship was found between SPN and accumulation mode MIPSD. The coefficient of variation (COV) of SPN emissions of particles >23 nm ranged from 8 to 26% for the UDDS and on-road tests.

Diet influences expression of autoimmune-associated genes and disease severity by epigenetic mechanisms in a transgenic mouse model of lupus.

OBJECTIVE: Lupus flares occur when genetically predisposed individuals encounter appropriate environmental agents. Current evidence indicates that the environment contributes by inhibiting T cell DNA methylation, causing overexpression of normally silenced genes. DNA methylation depends on both dietary transmethylation micronutrients and ERK-regulated DNA methyltransferase 1 (DNMT-1) levels. We used transgenic mice to study the effect of interactions between diet, DNMT-1 levels, and genetic predisposition on the development and severity of lupus. METHODS: A doxycycline-inducible ERK defect was bred into lupus-resistant (C57BL/6) and lupus-susceptible (C57BL/6 × SJL) mouse strains. Doxycycline-treated mice were fed a standard commercial diet for 18 weeks and then switched to a transmethylation micronutrient-supplemented (MS) or -restricted (MR) diet. Disease severity was assessed by examining anti-double-stranded DNA (anti-dsDNA) antibody levels, the presence of proteinuria and hematuria, and by histopathologic analysis of kidney tissues. Pyrosequencing was used to determine micronutrient effects on DNA methylation. RESULTS: Doxycycline induced modest levels of anti-dsDNA antibodies in C57BL/6 mice and higher levels in C57BL/6 × SJL mice. Doxycycline-treated C57BL/6 × SJL mice developed hematuria and glomerulonephritis on the MR and standard diets but not the MS diet. In contrast, C57BL/6 mice developed kidney disease only on the MR diet. Decreasing ERK signaling and methyl donors also caused demethylation and overexpression of the CD40lg gene in female mice, consistent with demethylation of the second X chromosome. Both the dietary methyl donor content and the duration of treatment influenced methylation and expression of the CD40lg gene. CONCLUSION: Dietary micronutrients that affect DNA methylation can exacerbate or ameliorate disease in this transgenic murine lupus model, and contribute to lupus susceptibility and severity through genetic-epigenetic interactions.

Serum proteins reflecting inflammation, injury and repair as biomarkers of disease activity in ANCA-associated vasculitis.

OBJECTIVE: To identify circulating proteins that distinguish between active anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) and remission in a manner complementary to markers of systemic inflammation. METHODS: Twenty-eight serum proteins representing diverse aspects of the biology of AAV were measured before and 6 months after treatment in a large clinical trial of AAV. Subjects (n=186) enrolled in the Rituximab in ANCA-Associated Vasculitis (RAVE) trial were studied. Erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) levels were available for comparison. The primary outcome was the ability of markers to distinguish severe AAV (Birmingham Vasculitis Activity Score for Wegener's granulomatosis (BVAS/WG)≥3 at screening) from remission (BVAS/WG=0 at month 6), using areas under receiver operating characteristic (ROC) curve (AUC). RESULTS: All subjects had severe active vasculitis (median BVAS/WG=8) at screening. In the 137 subjects in remission at month 6, 24 of the 28 markers showed significant declines. ROC analysis indicated that levels of CXCL13 (BCA-1), matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinases-1 (TIMP-1) best discriminated active AAV from remission (AUC>0.8) and from healthy controls (AUC>0.9). Correlations among these markers and with ESR or CRP were low. CONCLUSIONS: Many markers are elevated in severe active AAV and decline with treatment, but CXCL13, MMP-3 and TIMP-1 distinguish active AAV from remission better than the other markers studied, including ESR and CRP. These proteins are particularly promising candidates for future studies to address unmet needs in the assessment of patients with AAV.

Development of brake activity measurement method for heavy-duty vehicles.

Tailpipe PM (particulate matter) emissions have been reduced due to decades of tightening regulations, however non-tailpipe PM emissions are not regulated and are expected to become a significant source of traffic-related PM emissions. Previous studies have focused on emission measurement from laboratory and track tests. Their findings suggest brake wear PM emission rates are dependent on brake activity. Therefore, it is important to characterize brake emissions by first understanding the real-world brake activity from many different vehicle vocations and driving conditions. The goal of the current study is to establish a test method and analysis for brake activity measurements of heavy-duty vehicles. In this study, brake fluid pressure and brake pad temperature were measured for a heavy-duty vehicle during chassis and on-road driving tests. The chassis tests consisted of the Central Business District (CBD) cycle representative of a repetitive stop-and-go driving pattern of a bus, and the Urban Dynamometer Driving Schedule (UDDS) cycle representative of urban driving conditions of heavy-duty vehicles. The on-road tests consisted of a local Riverside City route focused on urban roads at low vehicle speeds with frequent braking, while the second route from Riverside City to Victorville focused on highway driving and downhill braking. The brake pad temperature of the triplicate CBD cycle gradually increased linearly with a slope of 2.3°C/min and the temperature per kinetic energy lost during braking increased by 2.3 × 10-5°C/J for the CBD cycle. The UDDS cycles had the largest kinetic energy loss between 3.2 × 103 to 3.0 × 105 J in the histogram. The local Riverside city route brake temperature increased by 2.0°C/min. The kinetic energy loss for the on-road tests were one order of magnitude larger than that of the dynamometer tests due to brake events occurring under higher speeds.Implications: The non-tailpipe source contributions to traffic related particulate matter (PM) emissions have surpassed that of tailpipe emissions. The results of this work provide a measurement method to obtain brake activity information for a heavy-duty vehicle, which is critical estimating emission inventory accurately.

Environmental exposure, estrogen and two X chromosomes are required for disease development in an epigenetic model of lupus.

Systemic lupus erythematosus (SLE) is an autoimmune disease primarily afflicting women. The reason for the gender bias is unclear, but genetic susceptibility, estrogen and environmental agents appear to play significant roles in SLE pathogenesis. Environmental agents can contribute to lupus susceptibility through epigenetic mechanisms. We used (C57BL/6xSJL)F1 mice transgenic for a dominant-negative MEK (dnMEK) that was previously shown to be inducibly and selectively expressed in T cells. In this model, induction of the dnMEK by doxycycline treatment suppresses T cell ERK signaling, decreasing DNA-methyltransferase expression and resulting in DNA demethylation, overexpression of immune genes Itgal (CD11a) and Tnfsf7 (CD70), and anti-dsDNA antibody. To examine the role of gender and estrogen in this model, male and female transgenic mice were neutered and implanted with time-release pellets delivering placebo or estrogen. Doxycycline induced IgG anti-dsDNA antibodies in intact and neutered, placebo-treated control female but not male transgenic mice. Glomerular IgG deposits were also found in the kidneys of female but not male transgenic mice, and not in the absence of doxycycline. Estrogen enhanced anti-dsDNA IgG antibodies only in transgenic, ERK-impaired female mice. Decreased ERK activation also resulted in overexpression and demethylation of the X-linked methylation-sensitive gene CD40lg in female but not male mice, consistent with demethylation of the second X chromosome in the females. The results show that both estrogen and female gender contribute to the female predisposition in lupus susceptibility through hormonal and epigenetic X-chromosome effects and through suppression of ERK signaling by environmental agents.

Maternal micronutrient supplementation suppresses T cell chemokine receptor expression and function in F1 mice.

Prenatal environmental exposures play a critical role in determining late-life chronic disease susceptibility. However, the mechanisms linking the in utero environment and disease development in the offspring are poorly understood. Recent investigations have confirmed a central pathogenic role of T cell chemokine receptors, particularly C-C chemokine receptor (CCR) 2 and CCR5, in chronic inflammatory conditions. This study was designed to determine the effect of a synthetic prenatal micronutrient supplementation (MS) diet rich in methionine pathway metabolites on the T cell chemokine system in F1 C57Bl/6 mice. Female mice were fed either an MS or control diet 3 wk prior to mating, during pregnancy, and lactation. At 4 wk of age, F1 mice were killed for experiments or were fed the standard NIH-31 diet and allowed to age. Food consumption, maternal weight gain, and litter size were similar in dams fed the control and MS diets. However, the F1 offspring of dams fed the MS diet were smaller in size (P < 0.001). T cells from the MS F1 offspring had global hypermethylation compared with control F1 offspring (P < 0.005), corresponding to lower T cell chemokine receptor expression [CCR2 (P < 0.001), CCR5 (P < 0.001), and C-x-C chemokine receptor 3 (P < 0.01)] and cytokine expression [TNFα (P < 0.05), IL-2 (P < 0.001), and IL-4 (P < 0.01)]. Reduced T cell chemokine receptor gene expression in MS F1 mice was associated with decreased chemotaxis in vitro to C-C chemokine ligand (CCL) 2 and C-X-C chemokine ligand 10 (P < 0.01) and in vivo to CCL2 (P < 0.01). Taken together, the results suggest that epigenetic alteration through prenatal diet manipulation reduces the response to proinflammatory signals in mice.

Is blood pressure reduction a valid surrogate endpoint for stroke prevention? An analysis incorporating a systematic review of randomised controlled trials, a by-trial weighted errors-in-variables regression, the surrogate threshold effect (STE) and the Biomarker-Surrogacy (BioSurrogate) Evaluation Schema (BSES).

BACKGROUND: Blood pressure is considered to be a leading example of a valid surrogate endpoint. The aims of this study were to (i) formally evaluate systolic and diastolic blood pressure reduction as a surrogate endpoint for stroke prevention and (ii) determine what blood pressure reduction would predict a stroke benefit. METHODS: We identified randomised trials of at least six months duration comparing any pharmacologic anti-hypertensive treatment to placebo or no treatment, and reporting baseline blood pressure, on-trial blood pressure, and fatal and non-fatal stroke. Trials with fewer than five strokes in at least one arm were excluded. Errors-in-variables weighted least squares regression modelled the reduction in stroke as a function of systolic blood pressure reduction and diastolic blood pressure reduction respectively. The lower 95% prediction band was used to determine the minimum systolic blood pressure and diastolic blood pressure difference, the surrogate threshold effect (STE), below which there would be no predicted stroke benefit. The STE was used to generate the surrogate threshold effect proportion (STEP), a surrogacy metric, which with the R-squared trial-level association was used to evaluate blood pressure as a surrogate endpoint for stroke using the Biomarker-Surrogacy Evaluation Schema (BSES3). RESULTS: In 18 qualifying trials representing all pharmacologic drug classes of antihypertensives, assuming a reliability coefficient of 0.9, the surrogate threshold effect for a stroke benefit was 7.1 mmHg for systolic blood pressure and 2.4 mmHg for diastolic blood pressure. The trial-level association was 0.41 and 0.64 and the STEP was 66% and 78% for systolic and diastolic blood pressure respectively. The STE and STEP were more robust to measurement error in the independent variable than R-squared trial-level associations. Using the BSES3, assuming a reliability coefficient of 0.9, systolic blood pressure was a B + grade and diastolic blood pressure was an A grade surrogate endpoint for stroke prevention. In comparison, using the same stroke data sets, no STEs could be estimated for cardiovascular (CV) mortality or all-cause mortality reduction, although the STE for CV mortality approached 25 mmHg for systolic blood pressure. CONCLUSIONS: In this report we provide the first surrogate threshold effect (STE) values for systolic and diastolic blood pressure. We suggest the STEs have face and content validity, evidenced by the inclusivity of trial populations, subject populations and pharmacologic intervention populations in their calculation. We propose that the STE and STEP metrics offer another method of evaluating the evidence supporting surrogate endpoints. We demonstrate how surrogacy evaluations are strengthened if formally evaluated within specific-context evaluation frameworks using the Biomarker- Surrogate Evaluation Schema (BSES3), and we discuss the implications of our evaluation of blood pressure on other biomarkers and patient-reported instruments in relation to surrogacy metrics and trial design.