Validation of Biomarkers of CVD Risk from Dried Blood Spots in Community-Based Research: Methodologies and Study-Specific Serum Equivalencies.

Dried blood spot (DBS) methodology offers significant advantages over venipuncture in studies of vulnerable populations or large-scale studies, including reduced participant burden and higher response rates. Uncertainty about the validity of cardiovascular disease (CVD) risk biomarkers remains a barrier to wide-scale use. We determined the validity of DBS-derived biomarkers of CVD risk versus gold-standard assessments, and study-specific, serum-equivalency values for clinical relevance of DBS-derived values. Concurrent venipuncture serum and DBS samples (n = 150 adults) were assayed in Clinical Laboratory Improvement Amendments-certified and DBS laboratories, respectively. Time controls of DBS standard samples were assayed single-blind along with test samples. Linear regression analyses evaluated DBS-to-serum equivalency values; agreement and bias were assessed via Bland-Altman plots. Linear regressions of venipuncture values on DBS-to-serum equivalencies provided R(2) values for total cholesterol, high-density lipoprotein cholesterol (HDL-C), and C-reactive protein (CRP) of 0.484, 0.118, and 0.666, respectively. Bland-Altman plots revealed minimal systematic bias between DBS-to-serum and venipuncture values; precision worsened at higher mean values of CRP. Time controls revealed little degradation or change in analyte values for HDL-C and CRP over 30 weeks. We concluded that DBS-assessed biomarkers represent a valid alternative to venipuncture assessments. Large studies using DBS should include study-specific serum-equivalency determinations to optimize individual-level sensitivity, the viability of detecting intervention effects, and generalizability in community-level primary prevention interventions.

Work stress, sleep deficiency, and predicted 10-year cardiometabolic risk in a female patient care worker population.

OBJECTIVES: The aim of this study was to investigate the longitudinal effect of work-related stress, sleep deficiency, and physical activity on 10-year cardiometabolic risk among an all-female worker population. METHODS: Data on patient care workers (n=99) was collected 2 years apart. Baseline measures included: job stress, physical activity, night work, and sleep deficiency. Biomarkers and objective measurements were used to estimate 10-year cardiometabolic risk at follow-up. Significant associations (P<0.05) from baseline analyses were used to build a multivariable linear regression model. RESULTS: The participants were mostly white nurses with a mean age of 41 years. Adjusted linear regression showed that having sleep maintenance problems, a different occupation than nurse, and/or not exercising at recommended levels at baseline increased the 10-year cardiometabolic risk at follow-up. CONCLUSIONS: In female workers prone to work-related stress and sleep deficiency, maintaining sleep and exercise patterns had a strong impact on modifiable 10-year cardiometabolic risk.

Dried blood spot collection of health biomarkers to maximize participation in population studies.

Biomarkers are directly-measured biological indicators of disease, health, exposures, or other biological information. In population and social sciences, biomarkers need to be easy to obtain, transport, and analyze. Dried Blood Spots meet this need, and can be collected in the field with high response rates. These elements are particularly important in longitudinal study designs including interventions where attrition is critical to avoid, and high response rates improve the interpretation of results. Dried Blood Spot sample collection is simple, quick, relatively painless, less invasive then venipuncture, and requires minimal field storage requirements (i.e. samples do not need to be immediately frozen and can be stored for a long period of time in a stable freezer environment before assay). The samples can be analyzed for a variety of different analytes, including cholesterol, C-reactive protein, glycosylated hemoglobin, numerous cytokines, and other analytes, as well as provide genetic material. DBS collection is depicted as employed in several recent studies.

Relationship of sleep deficiency to perceived pain and functional limitations in hospital patient care workers.

OBJECTIVE: Health care workers are at high risk of developing musculoskeletal symptoms and pain. This study tested the hypothesis that sleep deficiency is associated with pain, functional limitations, and physical limitations that interfere with work. METHODS: Hospital patient care workers completed a survey (79% response rate) including measures of health, sociodemographic, and workplace factors. Associations of sleep deficiency with pain, work interference due to this pain, and functional limitations were determined. RESULTS: Of 1572 respondents (90% women; mean age, 41 years), 57% reported sleep deficiency, 73% pain in last 3 months, 33% work interference, and 18% functional limitation. Sleep deficiency was associated with higher rates of pain, work interference, and functional limitation controlling for socioeconomic, individual, and workplace characteristics. CONCLUSIONS: Sleep deficiency is significantly associated with pain, functional limitation, and workplace interference, suggesting modifiable outcomes for workplace health and safety interventions.

Adverse metabolic consequences in humans of prolonged sleep restriction combined with circadian disruption.

Epidemiological studies link short sleep duration and circadian disruption with higher risk of metabolic syndrome and diabetes. We tested the hypotheses that prolonged sleep restriction with concurrent circadian disruption, as can occur in people performing shift work, impairs glucose regulation and metabolism. Healthy adults spent >5 weeks under controlled laboratory conditions in which they experienced an initial baseline segment of optimal sleep, 3 weeks of sleep restriction (5.6 hours of sleep per 24 hours) combined with circadian disruption (recurring 28-hour "days"), followed by 9 days of recovery sleep with circadian re-entrainment. Exposure to prolonged sleep restriction with concurrent circadian disruption, with measurements taken at the same circadian phase, decreased the participants' resting metabolic rate and increased plasma glucose concentrations after a meal, an effect resulting from inadequate pancreatic insulin secretion. These parameters normalized during the 9 days of recovery sleep and stable circadian re-entrainment. Thus, in humans, prolonged sleep restriction with concurrent circadian disruption alters metabolism and could increase the risk of obesity and diabetes.

Activity-based metagenomic screening and biochemical characterization of bovine ruminal protozoan glycoside hydrolases.

The rumen, the foregut of herbivorous ruminant animals such as cattle, functions as a bioreactor to process complex plant material. Among the numerous and diverse microbes involved in ruminal digestion are the ruminal protozoans, which are single-celled, ciliated eukaryotic organisms. An activity-based screen was executed to identify genes encoding fibrolytic enzymes present in the metatranscriptome of a bovine ruminal protozoan-enriched cDNA expression library. Of the four novel genes identified, two were characterized in biochemical assays. Our results provide evidence for the effective use of functional metagenomics to retrieve novel enzymes from microbial populations that cannot be maintained in axenic cultures.

Effects of lexical status and morphological complexity in masked priming: An ERP study.

Two masked priming experiments examined behavioural and event-related potential responses to simplex target words (e.g., flex) preceded by briefly presented, masked, derived word primes (flexible-flex), complex nonword primes formed by an illegal combination of the target word and a real suffix (flexify-flex), and simplex nonword primes formed by adding a nonsuffix word ending to the target (flexint-flex). Subjects performed a lexical decision task. Behavioural results showed that all prime types significantly facilitated target recognition. Priming effects were reflected in the electrophysiological data by reduced N250 and N400 amplitudes, and these priming effects were statistically equivalent for the three types of prime. The strong priming effects found with simplex primes in the present study, compared with prior research, are thought to be due to the combination of targets always being completely embedded in prime stimuli plus the reduced lexical inhibition that arises with nonword primes. In line with prior behavioural research, however, there was evidence for differential priming effects as a function of prime type in the N400 ERP component in Experiment 2, with greater priming effects for derived and pseudocomplex primes relative to simplex primes at lateral posterior electrode sites.

Aging is associated with myocardial insulin resistance and mitochondrial dysfunction.

Aging is associated with insulin resistance, often attributable to obesity and inactivity. Recent evidence suggests that skeletal muscle insulin resistance in aging is associated with mitochondrial alterations. Whether this is true of the senescent myocardium is unknown. Twelve young (Y, 4 years old) and 12 old (O, 11 years old) dogs, matched for body mass, were instrumented with left-ventricular pressure gauges, aortic and coronary sinus catheters, and flow probes on left circumflex artery. Before surgery, all dogs participated in a 6-wk exercise program. Dogs underwent measurements of hemodynamics and plasma substrates before and during a 2-h hyperinsulinemic-euglycemic clamp to measure whole body and myocardial glucose and nonesterified fatty acid uptake. Following the protocol, myocardial and skeletal samples were obtained to measure components of the insulin-signaling cascade and mitochondrial structure. There was no difference in plasma glucose (Y, 90 +/- 4 mg/dl; O, 87 +/- 4 mg/dl), but old dogs had higher (P < 0.02) nonesterified fatty acids (Y, 384 +/- 48 micromol/l; O, 952 +/- 97 micromol/l) and plasma insulin (Y, 39 +/- 11 pmol/l; O, 108 +/- 18 pmol/l). Old dogs had impaired total body glucose disposition (Y, 11.5 +/- 1 mg x kg(-1) x min(-1); O, 8.0 +/- 0.5 mg x kg(-1) x min(-1); P < 0.05) and insulin-stimulated myocardial glucose uptake (Y, 3.5 +/- 0.3 mg x min(-1) x g(-1); O, 1.8 +/- 0.3 mg x min(-1) x g(-1); P < 0.05). The impaired insulin action was associated with altered insulin signaling and glucose transporter (GLUT4) translocation. There were myocardial mitochondrial structural changes observed in association with decreased expression of uncoupling protein-3. Aging is associated with both whole body and myocardial insulin resistance, independent of obesity and inactivity, but involving altered mitochondrial structure and impaired cellular insulin action.

Fiber digestibility by the orangutan (Pongo abelii): in vitro and in vivo.

Limited nutritional information exists on diets of free-ranging orangutans, Pongo abelii and P. pygmaeus. Although they are classified as frugivores, the chemical composition of their diet and their gastrointestinal anatomy suggest that they rely on fiber fermentation for a substantial portion of energy. However, the extent to which they can ferment fiber is not known. Continuous culture systems, inoculated with orangutan fecal bacteria, were established to determine the fiber-digesting capacity of orangutan hindgut microflora. The cultures received one of four treatments: soybean hulls, ground corncobs, corn starch, or no food. Neither dry matter nor neutral detergent fiber digestibilities differed significantly among treatments. However, neutral detergent fiber digestibilities were high for both the soybean hull (88.4%) and ground corncob (86.1%) treatments, indicating that the microflora had a strong fibrolytic capability. To determine whether the same fiber-degrading capacity occurred in vivo, two adult orangutans and one juvenile were fed four gel-matrix diets containing soybean hulls, ground corncobs, or ground primate biscuits. Neutral detergent fiber concentrations (dry matter basis) of the gel matrices were 52.9% with soybean hulls, 46.8% and 63.7% with ground corncobs, and 31.3% with ground primate biscuits. A fifth diet consisted of primate biscuits with 27.3% neutral detergent fiber (dry matter basis) and was considered the baseline diet. Neutral detergent fiber digestibility (74.5%) was greatest (P < 0.05) for the soybean hull gel diet and least (57.5% and 45.0%, respectively; P < 0.05) for the 63.7% neutral detergent fiber (dry matter basis) corncob gel diet and the baseline primate biscuit diet. Total volatile fatty acid concentrations in orangutan feces were not significantly different among diets; however, molar proportions of acetic, propionic, and butyric acid differed (P < 0.05) among diets. The results from both studies indicated that orangutans are capable of extensive fiber fermentation.

Fiber digestibility by black lemurs (Eulemur macaco macaco).

Black lemurs, Eulemur macaco, are classified as generalist feeders, consuming a proportion of fruits and leaves that varies with seasonal availability. It is hypothesized that black lemurs are capable of using neutral detergent fiber (NDF) as a source of energy through fiber fermentation in the cecum and large intestine. In captivity, they are typically fed a diet of commercially available primate biscuits and readily available produce, both of which are limited in NDE Digestibility trials were conducted on 14 black lemurs; 12 were housed in groups (four, three, three, and two) and 2 were individually housed. The lemurs were fed four manufactured feeds differing in fiber form and content. A commercially available primate biscuit, containing approximately 27% NDF, served as the control diet. The second diet contained the same primate biscuits, which were ground and then incorporated into a fiber-based gel matrix. The third and fourth diets were formulated using soybean hulls or ground corncobs as the fiber source, resulting in 53% and 47% NDF, respectively. Produce was added to the diet at 36% (dry matter basis). Dry matter digestibility differed significantly among all diets. Neutral detergent fiber digestibility was highest for the ground biscuit in gel and lowest for the two high-fiber gel diets, with biscuit digestibility values falling between the two extremes. The high-fiber gel diets were successful in increasing the NDF concentrations of the primate diets; however, black lemurs were capable of only limited fiber digestion.