Minimally invasive biomarkers to detect maternal physiological status in sow saliva and milk.

In this study, we aimed to validate existing plasma assays to measure biomarkers for maternal signalling in milk and saliva of lactating sows. These biological samples are minimally invasive to the animal and could give a physiological profile of maternal qualities available to their piglets. Sows were farrowed in a zero-confinement system, and their colostrum and milk samples were manually collected during naturally occurring let-downs (i.e. not induced) over the lactation period. Saliva sampling involved sows voluntarily accepting cotton buds to chew without restraint. Commercial kits designed for blood plasma were tested, and any modifications and results are given. We successfully measured total protein, cortisol, tumour necrosis factor-α (TNF-α) and oxytocin in pig milk and saliva and immunoglobulin G (IgG) in pig milk samples. We were unsuccessful at measuring relaxin and serotonin in these biological samples. We observed higher levels of biomarkers in milk than in saliva. The measurement of TNF-α in pig milk for the first time revealed increased levels with larger litters. This development will allow more detailed understanding of biomarkers in milk. There was also evidence that the minimally invasive technique of using saliva sampling did not interrupt natural oxytocin production around parturition.

Development and validation of a multiplex fluorescent microsphere immunoassay assay for detection of porcine cytokines.

Cytokines are cell signalling proteins that mediate a number of different physiological responses. The accurate measurement of cytokine profiles is important for a variety of diagnostic and prognostic scenarios in relation to animal health and welfare. Simultaneous quantification of cytokine profiles in a single sample is now possible using fluorescent microsphere immunoassays (FMIA). We describe the development and validation of a novel multiplex assay using the Bio-Plex® 200 system to quantify cytokines in five different porcine tissues (brain, placenta, synovial tissue and fluid, plasma). The cytokine profiles are both tissue, and research hypothesis, -dependent but include Interleukin-1beta (IL-1ß), Interleukin-4 (IL-4), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10) and Tumor necrosis factor (TNF-α). This methods paper is reported in two parts: the development of a FMIA for porcine tissues and validation of pre-treatment for optimal cytokine recovery in porcine brain, placenta, synovial tissue and plasma. Validation steps are critical in ensuring an assay is suitable for novel sample types. This technique advances traditional ELISAs by: •FMIA provides insight into the profiles of multiple porcine cytokines in certain situations (e.g. disease, parturition).•Use of the Bio-Plex® 200 system to investigate novel sample types, including brain, placenta and synovial tissue.•Multiplexing utilises a fraction of the sample volume compared with multiple ELISAs.

Multi-scale mechanics of granular solids from grain-resolved X-ray measurements.

This work discusses an experimental technique for studying the mechanics of three-dimensional (3D) granular solids. The approach combines 3D X-ray diffraction and X-ray computed tomography to measure grain-resolved strains, kinematics and contact fabric in the bulk of a granular solid, from which continuum strains, grain stresses, interparticle forces and coarse-grained elasto-plastic moduli can be determined. We demonstrate the experimental approach and analysis of selected results on a sample of 1099 stiff, frictional grains undergoing multiple uniaxial compression cycles. We investigate the inter-particle force network, elasto-plastic moduli and associated length scales, reversibility of mechanical responses during cyclic loading, the statistics of microscopic responses and microstructure-property relationships. This work serves to highlight both the fundamental insight into granular mechanics that is furnished by combined X-ray measurements and describes future directions in the field of granular materials that can be pursued with such approaches.

Investigating the evolving microstructure of lithium metal electrodes in 3D using X-ray computed tomography.

The growth of electrodeposited lithium microstructures on metallic lithium electrodes has prevented their use in rechargeable lithium batteries due to early performance degradation and safety implications. Understanding the evolution of lithium microstructures during battery operation is crucial for the development of an effective and safe rechargeable lithium-metal battery. This study employs both synchrotron and laboratory X-ray computed tomography to investigate the morphological evolution of the surface of metallic lithium electrodes during a single cell discharge and over numerous cycles, respectively. The formation of surface pits and the growth of mossy lithium deposits through the separator layer are characterised in three-dimensions. This has provided insight into the microstructural evolution of lithium-metal electrodes during rechargeable battery operation, and further understanding of the importance of separator architecture in mitigating lithium dendrite growth.

Quantifying Interparticle Forces and Heterogeneity in 3D Granular Materials.

Interparticle forces in granular materials are intimately linked to mechanical properties and are known to self-organize into heterogeneous structures, or force chains, under external load. Despite progress in understanding the statistics and spatial distribution of interparticle forces in recent decades, a systematic method for measuring forces in opaque, three-dimensional (3D), frictional, stiff granular media has yet to emerge. In this Letter, we present results from an experiment that combines 3D x-ray diffraction, x-ray tomography, and a numerical force inference technique to quantify interparticle forces and their heterogeneity in an assembly of quartz grains undergoing a one-dimensional compression cycle. Forces exhibit an exponential decay above the mean and partition into strong and weak networks. We find a surprising inverse relationship between macroscopic load and the heterogeneity of interparticle forces, despite the clear emergence of two force chains that span the system.

Identification and disruption of bacteria associated with sheep scab mites-novel means of control?

Psoroptes ovis mites, which cause psoroptic mange (sheep scab), were investigated to identify potential bacterial targets for endosymbiont control of sheep scab. In addition, transmission of bacteria to the sheep skin was investigated through the characterisation of bacteria present in P. ovis faecal trails and on the fleece environment by internal transcribed spacer (ITS) sequencing. A diverse range of bacteria was identified in addition to a potential endosymbiont candidate, Comamonas sp, which was detected in P. ovis by both ITS PCR and endosymbiont-specific PCR. Disruption of these bacteria within P. ovis, through the use of antibiotics, was explored; with significant reduction in mean mite survival when administered antibiotic diets compared with controls (LR4 = 23.12, P < 0.001). The antibiotic treatments also significantly affected the bacterial density (CFU/mite) within P. ovis, indicating that mite survival may be linked to the bacterial communities that they harbour. Although antibiotics are not suitable for practical application, these results suggest disrupting bacteria associated with P. ovis should be further investigated for novel control.

Progression and remission of urologic symptoms in the community: results of a longitudinal cluster analysis approach.

OBJECTIVE: To investigate the natural history of urologic symptom progression and remission by means of cluster analysis in a large, well-characterized cohort of men and women. METHODS: Cluster analysis was used to assign men and women to symptom clusters on the basis of the prevalence of 14 self-reported urologic symptoms. Data were analyzed from the Boston Area Community Health study at baseline (T1) and 5-year follow-up (T2). Cluster progression was defined as any change from a less symptomatic to a more symptomatic cluster; conversely, cluster remission was defined as movement from more symptomatic to less symptomatic clusters. Logistic regression models examined the association of sociodemographic, psychosocial, and health outcome measures with cluster progression and remission. RESULTS: Follow-up data were available from 4145 participants (1610 men; 2535 women). More than two thirds of men (69.2%) and women (68.2%) had stable symptom cluster assignments. Cluster progression occurred in 280 of 1610 (15.2%) men and 390 of 2535 (14.6%) women; cluster remission in 280 of 1610 (15.6%) men and 409 of 2535 (17.4%) women. In multivariate analyses, cluster progression was twice as common in men with incident depression (odds ratio = 2.43, 95% confidence interval 1.26-4.67) and 3 times more likely in men with ≥ 3 comorbidities at baseline. Urologic surgeries were uncommon in men and women and were not consistently related to cluster progression or remission. CONCLUSION: Urologic symptom clusters were relatively stable over a 5-year follow-up period for more than two thirds of men and women in our sample. Specific risk factors for progression were identified in men and women.

Trajectories of change in sagebrush steppe vegetation communities in relation to multiple wildfires.

Repeated perturbations, both biotic and abiotic, can lead to fundamental changes in the nature of ecosystems, including changes in state. Sagebrush steppe communities provide important habitat for wildlife and grazing for livestock. Fire is an integral part of these systems, but there is concern that increased ignition frequencies and invasive species are fundamentally altering them. Despite these issues, the majority of studies of fire effects in systems dominated by Artemisia tridentata wyomingensis have focused on the effects of single burns. The Arid Lands Ecology Reserve (ALE), in south-central Washington (U.S.A.), was one of the largest contiguous areas of sagebrush steppe habitat in the state until large wildfires burned the majority of it in 2000 and 2007. We analyzed data from permanent vegetation transects established in 1996 and resampled in 2002 and 2009. Our objective was to describe how the fires, and subsequent postfire restoration efforts, affected communities' successional pathways. Plant communities differed in response to repeated fire and restoration; these differences could largely be ascribed to the functional traits of the dominant species. Low-elevation communities, previously dominated by obligate seeders, moved furthest from their initial composition and were dominated by weedy, early-successional species in 2009. Higher-elevation sites with resprouting shrubs, native bunchgrasses, and few invasive species were generally more resilient to the effects of repeated disturbances. Shrub cover has been almost entirely removed from ALE, although there was some recovery where communities were dominated by resprouters. Bromus tectorum dominance was reduced by herbicide application in areas where it was previously abundant, but it increased significantly in untreated areas. Several resprouting species, notably Phlox longifolia and Poa secunda, expanded remarkably following competitive release from shrub canopies and/or abundant B. tectorum. Our results suggest that community dynamics can be understood through a state and transition model with two axes (shrub/grass and native/invasive abundance), although such models also need to account for differences in plant functional traits and disturbance regimes. We use our results to develop a conceptual model that will be validated with further research.

Relative contributions of multiple determinants to bone mineral density in men.

SUMMARY: Focus on individual risk factors for osteoporosis could allocate disproportionate attention to trivial relationships. We tested many recognized risk factors of osteoporosis for their association with bone mineral density (BMD) in multivariate models among men. Lean mass accounted for the most variance, with substantially less accounted for by demographic, strength, and health factors. INTRODUCTION: Osteoporosis in men has gained recognition as a public health problem, generating an interest in the search for risk factors. Isolation of individual risk factors could allocate disproportionate attention to relationships that may be of limited consequence. METHODS: The Boston Area Community Health/Bone (BACH/Bone) Survey is a population-based study of randomly selected community-dwelling men (age, 30-79 years). BMD and lean mass were measured by dual X-ray absorptiometry. Socioeconomic status, health history, and lifestyle factors were obtained via interview. Hormone levels and markers of bone turnover were obtained from non-fasting blood samples. Multivariate analyses measured relative contributions of covariates to femoral neck (hip), one-third distal radius (wrist), and lumbar spine BMD. RESULTS: Factors positively associated with BMD in multivariate models at the three sites were black race and appendicular lean mass. Asthma was consistently negatively associated. Various other risk factors also contributed significantly to each of the individual sites. R (2) values for the hip, wrist, and spine were 41%, 30%, and 24%, respectively. Lean mass accounted for the most explained variance at all three sites. CONCLUSIONS: These data emphasize the limitation of focusing on individual risk factors and highlight the importance of potentially modifiable lean mass in predicting BMD.

Litter and dead wood dynamics in ponderosa pine forests along a 160-year chronosequence.

Disturbances such as fire play a key role in controlling ecosystem structure. In fire-prone forests, organic detritus comprises a large pool of carbon and can control the frequency and intensity of fire. The ponderosa pine forests of the Colorado Front Range, USA, where fire has been suppressed for a century, provide an ideal system for studying the long-term dynamics of detrital pools. Our objectives were (1) to quantify the long-term temporal dynamics of detrital pools; and (2) to determine to what extent present stand structure, topography, and soils constrain these dynamics. We collected data on downed dead wood, litter, duff (partially decomposed litter on the forest floor), stand structure, topographic position, and soils for 31 sites along a 160-year chronosequence. We developed a compartment model and parameterized it to describe the temporal trends in the detrital pools. We then developed four sets of statistical models, quantifying the hypothesized relationship between pool size and (1) stand structure, (2) topography, (3) soils variables, and (4) time since fire. We contrasted how much support each hypothesis had in the data using Akaike's Information Criterion (AIC). Time since fire explained 39-80% of the variability in dead wood of different size classes. Pool size increased to a peak as material killed by the fire fell, then decomposed rapidly to a minimum (61-85 years after fire for the different pools). It then increased, presumably as new detritus was produced by the regenerating stand. Litter was most strongly related to canopy cover (r2 = 77%), suggesting that litter fall, rather than decomposition, controls its dynamics. The temporal dynamics of duff were the hardest to predict. Detrital pool sizes were more strongly related to time since fire than to environmental variables. Woody debris peak-to-minimum time was 46-67 years, overlapping the range of historical fire return intervals (1 to > 100 years). Fires may therefore have burned under a wide range of fuel conditions, supporting the hypothesis that this region's fire regime was mixed severity.

Assessment of aortic regurgitation by transesophageal color Doppler imaging of the vena contracta: validation against an intraoperative aortic flow probe.

OBJECTIVES: This study was performed to validate the accuracy of color flow vena contracta (VC) measurements of aortic regurgitation (AR) severity by comparing them to simultaneous intraoperative flow probe measurements of regurgitant fraction (RgF) and regurgitant volume (RgV). BACKGROUND: Color Doppler imaging of the vena contracta has emerged as a simple and reliable measure of the severity of valvular regurgitation. This study evaluated the accuracy of VC imaging of AR by transesophageal echocardiography (TEE). METHODS: A transit-time flow probe was placed on the ascending aorta during cardiac surgery in 24 patients with AR. The flow probe was used to measure RgF and RgV simultaneously during VC imaging by TEE. Flow probe and VC imaging were interpreted separately and in blinded fashion. RESULTS: A good correlation was found between VC width and RgF (r = 0.85) and RgV (r = 0.79). All six patients with VC width >6 mm had a RgF >0.50. All 18 patients with VC width <5 mm had a RgF <0.50. Vena contracta area also correlated well with both RgF (r = 0.81) and RgV (r = 0.84). All six patients with VC area >7.5 mm2 had a RgF >0.50, and all 18 patients with a VC area <7.5 mm2 had a RgF <0.50. In a subset of nine patients who underwent afterload manipulation to increase diastolic blood pressure, RgV increased significantly (34 +/- 26 ml to 41 +/- 27 ml, p = 0.042) while VC width remained unchanged (5.4 +/- 2.8 mm to 5.4 +/- 2.8 mm, p = 0.41). CONCLUSIONS: Vena contracta imaging by TEE color flow mapping is an accurate marker of AR severity. Vena contracta width and VC area correlate well with RgF and RgV obtained by intraoperative flow probe. Vena contracta width appears to be less afterload-dependent than RgV.

Comparison of vena contracta width by multiplane transesophageal echocardiography with quantitative Doppler assessment of mitral regurgitation.

Mitral regurgitation (MR) severity is routinely assessed by Doppler color flow mapping, which is subject to technical and hemodynamic variables. Vena contracta width may be less influenced by hemodynamic variables and has previously been shown to correlate with angiographic estimates of MR severity. This study was performed to compare mitral vena contracta width by multiplane transesophageal echocardiography (TEE) with simultaneous quantitative Doppler echocardiography in 35 patients with MR. The vena contracta width was measured at the narrowest portion of the MR jet as it emerged through the coaptation of the leaflets; it was identified in 97% of the patients. Vena contracta width correlated well with regurgitant volume (R2 = 0.81) and regurgitant orifice area (R2 = 0.81) by quantitative Doppler technique. A vena contracta width > or = 0.5 cm always predicted a regurgitant volume >60 ml and an effective regurgitant orifice area > or = 0.4 cm2 in all patients. A vena contracta width < or = 0.3 cm always predicted a regurgitant volume <45 ml and a regurgitant orifice area < or = 0.35 cm2. Thus, vena contracta width by multiplane TEE correlates well with mitral regurgitant volume and regurgitant orifice area by quantitative Doppler echocardiography and provides a simple method for the identification of patients with severe MR.

Detection of myocardial perfusion defects by contrast echocardiography in the setting of acute myocardial ischemia with residual antegrade flow.

Although myocardial contrast echocardiography accurately demarcates area at risk during total coronary occlusion, the ability of MCE to delineate area at risk in the presence of residual antegrade flow is unknown. We hypothesized that perfusion defects in myocardial segments supplied by severe coronary stenoses with residual antegrade flow could be detected by MCE using intravenous FS069. We studied 13 open-chest dogs using an intravenous injection of FS069 during intermittent harmonic imaging. Images were collected at baseline, during acute ischemia with residual antegrade flow, physiologic hyperemia (release of stenosis), and total coronary occlusion. Regional myocardial blood flow was assessed using colored microspheres. MCE risk area during acute ischemia with residual antegrade flow and total occlusion was planimetered and compared with pathologic risk area (area unstained by monastral blue). Background-subtracted peak videointensity in the risk area was assessed for all flow states. Regional myocardial blood flow confirmed expected flow states, being significantly greater during physiologic hyperemia (4.16 +/- 1.22 ml/min/g) than at baseline (0.71 +/- 0.19 ml/min/g) and significantly diminished during coronary stenosis with residual antegrade flow (0.20 +/- 0.16 ml/min/g) and total occlusion (0.09 +/- 0.06 ml/min/g; p < 0.0001). Myocardial risk area by MCE during coronary stenosis with residual antegrade flow correlated well with pathologic risk area determined by monastral blue staining (r = 0.86). Peak videointensity during coronary stenosis (111 +/- 27) was significantly less than at baseline (157 +/- 50) but greater than during total occlusion (81 +/- 34; p < 0.0001). In conclusion, intravenous FS069 in conjunction with intermittent harmonic imaging delineates area at risk in ischemic myocardium supplied by a coronary stenoses with residual antegrade flow. The presence of a perfusion defect on MCE does not necessarily imply that the coronary artery is totally occluded.

Safety and efficacy of QW7437, a new fluorocarbon-based echocardiographic contrast agent.

The aim of this study was to evaluate the safety and efficacy of QW7437, a new fluorocarbon-based transpulmonary myocardial echocardiographic contrast agent. QW7437 is an anionically charged 2% dodecafluoropentane emulsion molecule, similar to EchoGen, a contrast agent previously shown to be efficacious in providing myocardial opacification by means of venous injection. This new agent has theoretical potential to provide greater safety and efficacy as a result of (1) reduced adherence to the negatively charged vascular endothelium and (2) reduced microbubble coalescence. Myocardial contrast echocardiography was performed in 10 dogs to evaluate the safety and efficacy of this agent. QW7437 (0.05 ml/kg) was injected as an intravenous bolus during intermittent harmonic epicardial imaging. Hemodynamic variables including heart rate, blood pressure, pulmonary artery pressure, pulmonary capillary wedge pressure, cardiac output, and arterial blood gases were determined at baseline and serially after contrast administration. Left ventricular fractional area shortening the regional myocardial blood flow at rest and during hyperemia (adenosine 140 micrograms/kg/min) were measured before and after contrast echocardiography. QW7437 provided dense myocardial opacification which persisted for more than 2 minutes in all subjects. This prolonged contrast effect did not result in significant changes in any hemodynamic variables, left ventricular function, or myocardial blood flow. Future studies should address the potential of this agent for human use.

Does digoxin provide additional hemodynamic and autonomic benefit at higher doses in patients with mild to moderate heart failure and normal sinus rhythm?

OBJECTIVES: This study sought to examine the hemodynamic and autonomic dose response to digoxin. BACKGROUND: Previous studies have demonstrated an increase in contractility and heart rate variability with digitalis preparations. However, little is known about the dose-response to digoxin, which has a narrow therapeutic window. METHODS: Nineteen patients with moderate heart failure and a left ventricular ejection fraction < 0.45 were studied hemodynamically using echocardiography and blood pressure at baseline and after 2 weeks of low dose (0.125 mg daily) and 2 weeks of moderate dose digoxin (0.25 mg daily). Loading conditions were altered with nitroprusside at each study. Autonomic function was studied by assessing heart rate variability on 24-h Holter monitoring and plasma norepinephrine levels during supine rest. RESULTS: Low dose digoxin provided a significant increase in ventricular performance, but no further increase was seen with the moderate dose. Low dose digoxin reduced heart rate and increased heart rate variability. Moderate dose digoxin produced no additional increase in heart rate variability or reduction in sympathetic activity, as manifested by heart rate, plasma norepinephrine or low frequency/high frequency power ratio. In addition, we did not find that either low or moderate dose digoxin increased parasympathetic activity. CONCLUSIONS: We conclude that moderate dose digoxin provides no additional hemodynamic or autonomic benefit for patients with mild to moderate heart failure over low dose digoxin. Because higher doses of digoxin may predispose to arrhythmogenesis, lower dose digoxin should be considered in patients with mild to moderate heart failure.

Assessment of mitral regurgitation severity by Doppler color flow mapping of the vena contracta.

BACKGROUND: Although Doppler color flow mapping is widely used to assess the severity of mitral regurgitation (MR), a simple, accurate, and quantitative marker of MR by color flow mapping remains elusive. We hypothesized that vena contracta width by color flow mapping would accurately predict the severity of MR. METHODS AND RESULTS: We studied 80 patients with MR. Vena contracta width was measured in multiple views with zoom mode and nonstandard angulation to optimize its visualization. Flow volumes across the left ventricular outflow tract and mitral annulus were calculated by pulsed-Doppler technique to determine regurgitant volume. Effective regurgitant orifice area was calculated by dividing the regurgitant volume by the continuous-wave Doppler velocity-time integral of the MR jet. The cause of MR was ischemia in 24, dilated cardiomyopathy in 34 mitral valve prolapse in 12, endocarditis in 2, rheumatic disease in 2, mitral annular calcification in 1, and uncertain in 5. Regurgitant volumes ranged from 2 to 191 mL. Regurgitant orifice area ranged from 0.01 to 1.47 cm2. Single-plane vena contracta width from the parasternal long-axis view correlated well with regurgitant volume (r = .85, SEE = 20 mL) and regurgitant orifice area (r = .86, SEE = 0.15 cm2). Biplane vena contracta width from apical views correlated well with regurgitant volume (r = .85, SEE = 19 mL) and regurgitant orifice area (r = .88, SEE = 0.14 cm2). A biplane vena contracta width > or = 0.5 cm was always associated with a regurgitant volume > 60 mL and a regurgitant orifice area > 0.4 cm2. A biplane vena contracta width < or = 0.3 cm predicted a regurgitant volume < 60 mL and a regurgitant orifice area < 0.4 cm2 in 24 of 29 patients. No other parameter, including jet area, left atrial size, pulmonary flow reversal, or semiquantitative MR grade, correlated significantly with regurgitant volume or regurgitant orifice area in a multivariate analysis. CONCLUSIONS: Our results demonstrate that careful color flow mapping of the vena contracta of the MR jet provides a simple quantitative assessment of MR that correlates well with quantitative Doppler techniques.

An analysis of dilemmas posed by prehospital DNR orders.

This article briefly recounts the development of the prehospital do-not-resuscitate (DNR) order and indicates certain situations, such as a choking episode or a suicide attempt, in which the presence of a DNR order may provoke a moral dilemma for the emergency medical technician as to whether or not the patient should be treated. An ethical analysis of this question is performed and concludes that resuscitative treatment is ethical and mandatory.