Hourly Blood Samples Research Articles

The Timing of the Melatonin Onset and Phase Angle to Sleep Onset in Older Adults after Uncontrolled vs. Controlled Lighting Conditions.

The main aim of this study was to explore how melatonin onset timing and phase angle to bedtime in healthy older adults are impacted by prior light exposure. A total of 13 healthy older (ages 56-74) individuals were studied on two successive evenings. Prior to the first evening, the participants were in self-selected lighting conditions for the first 4-6 h of the day and then were in dim light (3 lux) until their scheduled bedtime. On the second day, individuals from Project A remained in the dim lighting conditions throughout the entire day but those in Project B were in more typical indoor lighting (~90 lux) throughout the day. On both evenings, hourly blood samples were collected and assayed for melatonin, and melatonin onset timing and phase angle to sleep onset was determined. Overall, melatonin onset was earlier and the phase angle was larger on Night 1 than on Night 2. In Project A there was no significant difference between melatonin onset on night 1 vs. night 2. However, in Project B melatonin onset was significantly later on Night 2 (in typical indoor lighting) than on Night 1 (in dim lighting). Our results suggest that in older people, uncontrolled bright light early in the day did not impact the timing of dim light melatonin onset (DLMO) when assessed later that same evening. However, in older adults, exposure to ordinary room light during melatonin phase assessment appeared to suppress melatonin, leading to a later observed time of melatonin onset, as has been reported previously for young adults.

The use of levothyroxine absorption tests in clinical practice.

Although levothyroxine (LT4) is a widely prescribed drug, over 30% of LT4-treated patients fail to achieve the recommended serum level of thyroid-stimulating hormone with a body-weight-based dose of LT4. An LT4 absorption test (LT4AT) is part of the work-up for confirming normal LT4 absorption or diagnosing malabsorption. We searched PubMed with the terms levothyrox*, L-T4, LT4, TT4, FT4, FT3, TT3, test, loading, uptake, absorp*, "absorb*, bioavailab*, bioequiv* malabsorb*, pseudomalabsorb*. 43 full-text publications were analyzed. The published procedures for LT4AT differ markedly in the test dose, formulation, test duration, frequency of blood collection, analyte (total thyroxine (TT4) or free thyroxine (FT4)), metric (absolute or relative peak or increment, or area under the curve) and the threshold for normal absorption. In a standardized LT4AT for routine use, the physician could: advise the patient to not consume food, beverages or medications on the morning of the test; administer 1000 µg of LT4 in the patient's usual formulation as the test dose; ensure that the patient is supervised throughout the LT4AT; perform a 4-hour test, with hourly blood samples; assay FT4; and consider that normal LT4 absorption corresponds to an FT4 increment of more than 0.40 ng/dL (5.14 pmol/L) or a TT4 increment of more than 6 μg/dL (77.23 nmol/L) for a test dose of at least 300 µg, or a percentage TT4 absorption of more than 60%. If the test indicates abnormal LT4 absorption, the physician can increase the LT4 dose, change the formulation or administration route, and/or refer the patient to a gastroenterologist.

A comparison of four methods to estimate dim light melatonin onset: a repeatability and agreement study

ABSTRACT Dim light melatonin onset (DLMO) is considered the most reliable circadian phase marker in humans. However, the methods to calculate it are diverse, which limits the comparability between studies. Given the key role of DLMO to diagnose circadian rhythm sleep-wake disorders and determine the optimal timing of chronotherapies, the establishment of clear and validated guidelines on the methodology to assess DLMO is very important. We performed a repeatability study (n = 31) and an agreement study (n = 62) in healthy young adults with hourly blood samples collected under dim light conditions (<8 lux) during a chronobiological protocol. We assessed the repeatability of DLMO with three different methods (fixed threshold, dynamic threshold and hockey stick) across two nights and assessed agreement of each method with the mean visual estimation made by four chronobiologists. Analyses included Bland-Altman diagrams, intraclass correlation coefficients and equivalence tests. The repeatability of the four methods across two nights ranged from good to perfect. The agreement study highlighted that the hockey stick showed equivalent or superior performance (ICC: 0.95, mean difference with visual estimation: 5 min) in healthy subjects compared to the dynamic and fixed thresholds. Thanks to its objective nature, the hockey stick method may provide better estimates than the mean of the visual estimations of several raters. These findings suggest that the hockey stick method provides the most reliable estimate of DLMO within the tested methods and should be considered for use in future studies.

Physiological circadian rhythm of cardiac myosin-binding protein C (cMyC) and cardiac troponin

Abstract Introduction Cardiac myosin-binding protein C (cMyC) is a novel protein biomarker of myocardial injury, with a promising role in the triage and risk stratification of patients with cardiac disease. Understanding the physiological diurnal oscillation of cMyC and cardiac troponin is important for the interpretation of single and serial measurements within the biomarker-assisted triage and risk stratification algorithms. Purpose In this study, we aim to assess and compare the physiological diurnal oscillation of cMyC and cardiac troponin cTnT and cTnI. Method Twenty-six consecutive hourly blood samples were drawn between 08.30 am and 09.30 am (+1 day) from normotensive 24 individuals without a recent history of acute myocardial infarction, for the measurement of cMyC, cardiac troponin T (Roche hs-cTnT) and I (Abbott hs-cTnI). Fitted cosinor sine regression model (with R, version 3.6.1) was used to assess the presence and significance of circadian oscillation of the biomarker, and to estimate the respective amplitude and acrophase (the time of peak activity). Amplitude and acrophase were compared across the biomarkers that exhibited significant circadian rhythm. Results Mean age was 72±7. 79% of participants (n=19) were men. All participants were free from renal disease. On population-mean cosinor analysis, hs-cTnI exhibited random diurnal oscillation, whereas significant circadian rhythm was detected for cMyC and hs-cTnT (p=0.015 and &amp;lt;0.001, respectively) (Figure 1). The circadian rhythm of cMyC is characterised by gradually increasing concentrations from early afternoon until early morning (acrophase 03:03 am, 95% CI 01:54–04:26 am) compared to hs-cTnT concentrations which exhibits delayed increase and a later peak (acrophase, 08:01, 95% CI 07:10–08:51 am), p=0.028 for acrophase difference (Figure 1). Diurnal rhythm remained significant after correction for possible posture-induced changes in plasma volume. To allow direct comparison between amplitudes, the measurements of cMyC and hs-TnT were normalised to the respective 08:30 am value, re-fitted cosinor model did not show significant difference between the amplitudes (amplitude ng/L, 0.12, 95% CI 0.07–0.15 vs 0.11, 95% CI 0.08–0.12, for normalised cMyC vs hs-cTnT, respectively; p=0.67). Conclusion Significant circadian rhythm exists for cMyC and hs-cTnT, with 5-hours phase difference between the two biomarkers (cMyC ahead of hs-cTnT). The cause of this rhythmic variation is unknown, but the phase difference is consistent with the previously described disparity in the release of cMyC and cTnT after iatrogenic myocardial injury, raising the possibility of an underlying diurnal variation in myocardial vulnerability. Studies are required to assess the impact of this physiological phenomenon on the performance of the biomarkers within unadjused diagnostic algorithms Funding Acknowledgement Type of funding sources: Foundation. Main funding source(s): British Heart FoundationStichting de Weijerhorst

Continuous Glucose Monitoring in the Management of Neonates With Persistent Hypoglycemia and Congenital Hyperinsulinism.

BackgroundPersistent hypoglycemia is common in the newborn and is associated with poor neurodevelopmental outcome. Adequate monitoring is critical in prevention, but is dependent on frequent, often hourly blood sampling. Continuous glucose monitoring (CGM) is increasingly being used in children with type 1 diabetes mellitus, but use in neonatology remains limited. We aimed to introduce real-time CGM to provide insights into patterns of dysglycemia and to support the management of persistent neonatal hypoglycemia.MethodsThis is a single-center retrospective study of real-time CGM use over a 4-year period in babies with persistent hypoglycemia.ResultsCGMs were inserted in 14 babies: 8 term and 6 preterm infants, 9 with evidence of congenital hyperinsulinism (CHI). A total of 224 days of data was collected demonstrating marked fluctuations in glucose levels in babies with CHI, with a higher sensor glucose SD (1.52 ± 0.79 mmol/L vs 0.77 ± 0.22 mmol/L) in infants with CHI compared with preterm infants. A total of 1254 paired glucose values (CGM and blood) were compared and gave a mean absolute relative difference of 11%.ConclusionCGM highlighted the challenges of preventing hypoglycemia in these babies when using intermittent blood glucose levels alone, and the potential application of CGM as an adjunct to clinical care.

Unacylated ghrelin, leptin, and appetite display diurnal rhythmicity in lean adults.

Constant routine and forced desynchrony protocols typically remove the effects of behavioral/environmental cues to examine endogenous circadian rhythms, yet this may not reflect rhythms of appetite regulation in the real world. It is therefore important to understand these rhythms within the same subjects under controlled diurnal conditions of light, sleep, and feeding. Ten healthy adults (9 M/1 F, means ±SD: age, 30 ± 10 yr; body mass index, 24.1 ± 2.7 kg·m-2) rested supine in the laboratory for 37 h. All data were collected during the final 24 h of this period (i.e., 0800-0800 h). Participants were fed hourly isocaloric liquid meal replacements alongside appetite assessments during waking before a sleep opportunity from 2200 to 0700 h. Hourly blood samples were collected throughout the 24-h period. Dim light melatonin onset occurred at 2318 ± 46 min. A diurnal rhythm in mean plasma unacylated ghrelin concentration was identified (P = 0.04), with the acrophase occurring shortly after waking (0819), falling to a nadir in the evening with a relative amplitude of 9%. Plasma leptin concentration also exhibited a diurnal rhythm (P < 0.01), with the acrophase occurring shortly after lights-out (0032 h) and the lowest concentrations at midday. The amplitude for this rhythm was 25%. Diurnal rhythms were established in all dimensions of appetite except for sweet preference (P = 0.29), with both hunger (2103 h) and prospective food consumption (1955 h) reaching their peak in the evening before falling to their nadir shortly after waking. Under controlled diurnal conditions, simultaneous measurement of leptin, unacylated ghrelin, and subjective appetite over a 24-h period revealed rhythmicity in appetite regulation in lean, healthy humans.NEW & NOTEWORTHY Simultaneous assessment of subjective appetite, unacylated ghrelin, and leptin was carried out over a continuous 37-h protocol for the first time under conditions of controlled light, sleep, and feeding in healthy, lean adults. Rhythms were observed in unacylated ghrelin, leptin, and components of subjective appetite, such as hunger, prospective consumption, and fullness. Concurrent measurement of rhythms in these variables is important to fully understand the temporal relationships between components of appetite as well as the influence of diurnal factors such as sleep, light, and feeding.

Concentrations of progesterone and a PGF2α metabolite during the interovulatory interval compared to the corresponding days of pregnancy in mares

The concentrations of progesterone (P4) and a metabolite of PGF2α (PGFM) in mares were compared between the interovulatory interval (IOI; n = 8) and the corresponding days of pregnancy (n = 9). In daily blood samples, P4 increased between the day of ovulation (Day 0) and ∼Day 6 and then gradually decreased until the beginning of luteolysis in the IOI group. Before the beginning of luteolysis, there were no significant differences in P4 concentrations between the IOI and early pregnancy. In the IOI, PGFM concentration on the day before the beginning of luteolysis began to increase (P < 0.04) and reached a maximum mean (42.9 ± 11.6 pg/mL) on Day 14. In pregnancy, a novel increase in PGFM occurred from Day 12 to a maximum mean on Day 15 (16.7 ± 3.1 pg/mL). Daily PGFM concentrations were not different between the two groups until the increase just before luteolysis in the IOI. During 8-h sessions of hourly blood sampling, the mean and maximum PGFM concentrations were significantly greater in IOI than in pregnancy for each 8-h session on Days 13, 14, and 15. The minimum was not different between groups on any day. Pulses of PGFM were identified by coefficient of variation during the hourly 8-h sessions on day-sets of Days 4–7, 9–11, and 13–16. Despite the PGFM increase in daily samples between Days 12 and 15 of pregnancy, the amplitude and peaks of CV-identified pulses did not differ in the pregnant mares among the three day-sets. The pulses were similarly small for day-sets 4–7 and 9–11 in the IOI and for all day-sets in pregnancy (eg, amplitude on Days 13–16: 43.4 ± 15.6 pg/mL vs 5.4 ± 1.1 pg/mL for IOI vs pregnancy). Hypothesis 1 was not supported that daily PGFM concentrations in an IOI increase at the intersection between the end of the rapid P4 increase and the gradual P4 decrease. Hypothesis 2 was supported that pregnant mares have low amplitude PGFM pulses during the days of the high amplitude pulses at luteolysis in the IOI.

Measuring Oral Vitamin B12 Bioavailability Using [13C]-cyanocobalamin in Humans

Abstract Objectives To biosynthesize a stable-isotope labeled [13C]-cyanocobalamin and to measure its bioavailability in humans. Methods Salmonella enterica was precultured in Luria-Bertani medium and single-cell colonies were grown in No carbon E medium, supplemented with [13C]-ethanolamine as a sole-carbon source along with other precursors (dicyanocobinamide and dimethylbenzimidazole) and incubated for 48 h at 30°C and centrifuged. The pellets were resuspended in methanol/sodium cyanide and incubated overnight and extracts were purified by HPLC. The peaks corresponding to standard cyanocobalamin were collected and characterized by MALDI-MS. After animal testing, healthy young subjects were orally administered with [13C]-cyanocobalamin in a protocol that included hourly blood sampling for 12 h post-dose, and daily sampling for the next 5 days, to characterize its kinetics at a higher dose than the daily requirement (∼10x, n = 4, 2 males and females) and its appearance was modeled using a two-compartment model with early and late absorption phases. The same model was applied for lower dose at daily requirement (∼2.5 μg, n = 11 males), with sampling for 12 h post-dose. In addition, the effect of co-ingested food (n = 1, male) and the effect of parenteral replenishment of existing body vitamin B12 stores by intramuscular injection of 1 mg hydroxocobalamin, (n = 3, males) on bioavailability was also studied. The appearance of tracer in plasma was quantified by Q-Exactive-Orbitrap-MS. Results Biosynthesized [13C]-cyanocobalamin was labeled with up to 7–13[C] atoms and at a daily requirement dose (∼2.5 μg), its mean bioavailability was 50.4 ± 8.2%, but this was also dependent on the existing body store of vitamin B12. Two-weeks after replenishing the body stores, bioavailability increased by 1.5-fold (from 45.6 ± 0.02% to 67.8 ± 0.06%). At a higher dose (∼20 μg), its bioavailability was 6.0 ± 1.2% but the absolute amount absorbed was similar to that with the low dose. Conclusions A novel stable isotope-based vitamin B12 bioavailability assessment is described at low and high doses. This has implications toward rationalizing dosages used in clinical supplementation and public health fortification programs. Funding Sources Department of Biotechnology, India (to SD and AVK) and by the Wellcome Trust/DBT India Alliance Margdarshi Fellowship [No. IA/M/14/1/501,681] awarded to AVK.

Some Negative Effects of Heat Stress in Feedlot Heifers May Be Mitigated via Yeast Probiotic Supplementation.

This study was designed to determine if supplementation of a combination live yeast and yeast cell wall product in feed could mitigate the negative impacts associated with heat stress (HS). Crossbred, phenotypically similar beef heifers (n = 32; BW = 385 ± 43 kg) were fed a standard finishing ration without (CON) or with a combination of a live yeast (1.5 g/hd/d) and yeast cell wall product (2.5 g/hd/d; YEAST; Phileo Lesaffre Animal Care, Milwaukee, WI). After 50 d of supplementation, heifers were transported to an environmentally-controlled facility and placed in individual bleeding stalls after indwelling jugular catheters and vaginal temperature (VT) loggers were inserted. Heifers were kept in thermoneutral (TN) conditions for 48 h [temperature-humidity index (THI) ~67; d 1–2] then were subjected to HS for 4 d (peak THI ~80; d 3–6). From d 2–6, hourly blood samples were collected for serum isolation from 1400 to 1800 h and again from 2200 to 0200 h which represented the daily targeted peak and nadir of THIs. A whole blood sample was collected twice daily at 1400 and 2200 h for complete blood counts (CBC). There was no difference in BW (P = 0.14) or ADG (P = 0.53) between the treatments during HS. Yeast-supplemented heifers exhibited reduced VT during HS compared to CON heifers (P < 0.01). There was no difference in water intake during the TN phase (P = 0.25); however, YEAST heifers consumed more water/h (P < 0.01) and had increased drinking bouts (P < 0.01) during HS compared to CON heifers. Respiration rates (RR) did not differ (P = 0.21) during TN, but YEAST heifers tended (P = 0.09) to have decreased RR during HS compared to CON heifers. There were no differences between treatments when evaluating CBC parameters (P ≥ 0.10). There was a tendency (P = 0.08) for greater cortisol in the CON than YEAST heifers during HS; however, glucose (P = 0.38) and NEFA (P = 0.70) concentrations did not differ. In summary, supplementation of live yeast and yeast cell wall products to feedlot heifers may mitigate some of the negative effects associated with HS in feedlot cattle as observed in decreased RR and VT and increased water intake.

HPA axis in psychotic major depression and schizophrenia spectrum disorders: Cortisol, clinical symptomatology, and cognition

The Hypothalamic Pituitary Adrenal (HPA) axis has been implicated in the pathophysiology of a variety of mood and cognitive disorders. Neuroendocrine studies have demonstrated HPA axis overactivity in major depression, a relationship of HPA axis activity to cognitive performance, and a potential role of HPA axis genetic variation in cognition. In schizophrenia differential HPA activity has been found, including higher rates of non-suppression to dexamethasone challenge and higher salivary cortisol levels, which have been a premonitory risk factor for conversion to psychosis in adolescents at risk for developing schizophrenia. The present study investigated the simultaneous roles HPA axis activity and clinical symptomatology play in poor cognitive performance. Patients with major depression with psychosis (PMD) or schizophrenia spectrum disorder (SCZ) and healthy controls (HC) were studied. All participants underwent a diagnostic interview and psychiatric ratings, a comprehensive neuropsychological battery, and overnight hourly blood sampling for cortisol. Cognitive performance did not differ between the clinical groups, though they both performed more poorly than the HC's across a variety of cognitive domains. Across all subjects, cognitive performance was negatively correlated with higher cortisol, and PMD patients had higher evening cortisol levels than did SCZ and HCs. Cortisol and clinical symptoms, as well as age, sex, and antipsychotic use predicted cognitive performance. Diathesis stress models and their links to symptomatology, cognition, and HPA function are discussed.

Evaluation of in vitro hemolysis and platelet activation of a newly developed maglev LVAD and two clinically used LVADs with human blood.

In vitro hemolysis testing remains one of the most important performance measures to judge the hemocompatibility of a left ventricular assist device (LVAD). Clinically relevant operating conditions and appropriate testing blood are essential to infer in vitro data for potential clinical use. This in vitro study was carried out to evaluate and compare the hemolytic performance of a newly developed magnetically levitated (maglev) LVAD (CH-VAD) with two clinically used LVADs (HVAD and HeartMate II (HMII)) using fresh human blood. A small volume (~300 mL) in vitro circulating flow loop was constructed with a LVAD generated flow of 4.5 L/min at the nominal or reported clinical operating speed for each LVAD. The blood was circulated in the loop for 4 hours with samples drawn at baseline and hourly. Plasma-free hemoglobin (PFH) concentrations in the hourly blood samples were determined with spectrophotometry. Normalized index of hemolysis (NIH) was calculated to compare the hemolytic performance of the CH-VAD and the two reference LVADs. Platelet activation was measured with flow cytometry. The experimental test for each device was repeated at least 7 times. The data from this study showed that all the three LVADs generated very low hemolysis (NIH <0.01 g/100L). The CH-VAD was found to have a significantly lower NIH value (0.00135 ± 0.00032 g/100L) compared to the HVAD (0.00525 ± 0.00183 g/100L) and the HMII (0.00583 ± 0.00182 g/100L). No statistically significant difference in device-generated hemolysis was found between the HVAD and the HMII. The level of platelet activation induced by the CH-VAD is significantly lower than those by the HVAD and the HMII. The data suggest that the shear-induced hemolysis and platelet activation of the CH-VAD are acceptable relative to the two LVADs currently in clinical use.

Endogenous and exogenous effects of PGF2α during luteolysis in mares

An inhibitor of PGF2α biosynthesis (flunixin meglumine, FM) was used to study the role of endogenous PGF2α on the luteolytic effect of exogenous PGF2α in mares. A 2-h infusion of PGF2α at a constant rate (total dose, 0.1 mg) on Day 10 (ovulation = Day 0) was used to mimic the maximal concentrations of a spontaneous pulse of a PGF2α metabolite (PGFM). Treatment with FM (1.7 mg/kg) was done 1 h before and 5 h after the start of PGF2α infusion. In hourly blood samples beginning 1 h before the start of PGF2α infusion, progesterone decreased (P < 0.05) similarly by 5 h in each of the PGF2α and PGF2α+FM groups but not in the controls (n = 5). In a study of spontaneous luteolysis, the same FM dose was given every 6 h from Day 13 until Day 17 or earlier if CL regression was indicated by an 80% decrease in luteal blood-flow signals. Blood was sampled for progesterone assay each day and 8 h of hourly blood sampling was done each day to characterize PGFM concentrations and pulses. Progesterone (P4) was lower (P < 0.05) in controls than in an FM group (n = 7) by Day 15. Luteolysis (P4 < 1 ng/mL) ended on Days 14–19 in individual controls. In contrast, luteolysis did not end until after Day 20 in 4 of 7 FM-treated mares. In the three mares with completion of luteolysis before Day 20 in the FM group, the interval from beginning to end of luteolysis was longer (P < 0.02) (4.5 ± 0.6 days) than in the controls (3.0 ± 0.4 days). During 8-h sessions of hourly blood sampling on Day 14, concentration of PGFM was significantly lower in the FM group for the minimal, mean, and maximal per session. Pulses of PGFM were identified by a CV methodology on each day in 7 of 7 and 3 of 7 mares in the controls and FM group, respectively. The four FM-treated mares without a CV-identified pulse were the four mares in which luteolysis did not occur before Day 20. In mares with detected pulses, PGFM was lower at each nadir and at the peak (86% lower) in the FM group than in controls, but the interval between nadirs or base of a pulse was not different between groups. Hypothesis 1 that endogenous PGF plays a role in the luteolytic effect of exogenous PGF2α was not supported. Hypothesis 2 that an inhibitor of PGF2α biosynthesis prevented or minimized the prominence of PGFM pulses and increased the frequency of persistent CL was supported.

Leptin and HPA axis activity in diabetic rats: Effects of adrenergic agonists

Type I Diabetes (T1D) is associated with reduced leptin levels and increased stress axis activity marked by elevations in norepinephrine (NE) levels in the paraventricular nucleus (PVN) of the hypothalamus. We hypothesized that leptin suppresses stress axis activity in T1D through central and peripheral mechanisms. In the first experiment, adult male Sprague Dawley rats were implanted with a cannula in the PVN and randomly divided into a non-diabetic group treated with vehicle (n = 6) and a diabetic group treated with streptozotocin (n = 13). Food intake and water intake was measured for 14 days. On the last day, a subset of diabetic rats were treated with 500 µg of leptin i.p. Rats were subjected to push-pull perfusion of the PVN and hourly blood sampling for 5 h. In the next experiment, diabetic rats were treated either with an alpha-2 adrenergic agonist, clonidine (CLON), or a beta adrenergic agonist isoproterenol (ISO), to reverse the effects of leptin. Rats were subjected to push pull perfusion and hourly blood sampling. In experiment 1, T1D increased food intake, water intake, NE release in the PVN and circulating CS levels. Leptin treatment decreased NE release modestly but produced a robust reduction in corticosterone (CS) levels. In experiment 2, CLON but not ISO was able to reverse the effect of leptin on NE levels in the PVN, however, both agonists were capable of blocking leptin’s effects on circulating CS. These results suggest that in diabetic rats, the sensitivity of the hypothalamus to beta adrenergic agonists is altered, while the adrenals remain sensitive to both alpha and beta adrenergic agonists.

0117 Impact of Light Exposure during Sleep on Cardiometabolic Function

Artificial light exposure is increasingly widespread, particularly at night. Light exposure at night is known to impact circadian rhythms such as melatonin and sleep, and disturbances in these rhythms have been shown to affect cardiometabolic function. As such, the aim of this study was to test the hypothesis that light exposure at night during sleep negatively impacts cardiometabolic outcomes, possibly via disruptions to sleep architecture and melatonin profiles. Twenty healthy adults 18–40 years of age were randomized into Dark-Dark (DD) or Dark-Light (DL) groups and run in parallel for a three day and two night stay. Participants had 8 hours of sleep opportunity each night starting at habitual bedtime determined from one week of actigraphy with sleep diary. The DL group (n=10, 2 males, ages 26.61 ± 4.64 years, BMI 23.25 ± 3.94 kg/m^2) slept in the dark < 3 lux on Night 1 and slept in overhead room light of 100 lux on Night 2, while the DD group (n=10, 4 males, ages 26.78 ± 5.15 years, BMI 24.25 ± 3.71 kg/m^2) slept in the dark <3 lux on both Nights 1 and 2. Overnight polysomnography and hourly blood sampling for melatonin were collected on both nights. Oral glucose tolerance tests were performed on both mornings following sleep in the dark or 100 lux of light. Repeated measures ANOVA were performed between groups (DL vs. DD) with Day (Day 1 vs Day 2) and Time of Day as within-subject factors. Homeostatic model assessment of insulin resistance values were significantly higher (p<0.05) in the morning following sleep in the light (DL group) compared to sleep in the dark (DD group). This effect was primarily due to increased insulin levels for DL compared to DD group, for which there was a trend for group by day effect (p<0.09). A single night of light exposure during sleep acutely impacts insulin resistance, and chronic overnight exposure may have long-term effects on metabolic function. This research was supported National Institutes of Health grants 5T32HL7909, P01AG11412, and 8UL1TR000150-05.

The effect of aspirin on prostaglandin F2α secretion in lactating dairy cows

Early embryonic loss may be related to untimely secretion of prostaglandin F2α. The objective of this study was to examine the effect of aspirin, a nonsteroidal anti-inflammatory drug (NSAID), on prostaglandin F2α secretion and luteal function in lactating dairy cows by characterizing plasma prostaglandin metabolites (PGFM) and progesterone (P4) during the luteal phase. Estrous cycles were synchronized in 23 lactating Holstein cows using a 5-d CIDR-Ovsynch protocol. Transrectal ultrasonography was performed to confirm ovulation and formation of a corpus luteum. On d 14, after after the second injection of gonadotropin-releasing hormone, cows were assigned randomly to receive aspirin (n = 11, 46.65 g/dose) orally on d 14 (2×) and d 15 (1×), 12 h apart, or no aspirin (n = 12, control). On d 15, hourly blood samples were collected for PGFM, and daily blood samples were collected (d 14 to 22) for P4 concentrations. Mean P4 concentrations before treatment (d 14) did not differ between groups. There was an effect of treatment and treatment × time on PGFM (P < 0.05). Mean PGFM concentrations decreased (P < 0.05) after treatment and remained low for 11 h after last aspirin dose, whereas in the control PGFM remained unchanged. For cows that underwent luteolysis (n = 16), P4 concentrations were greater (P = 0.05), and the days to luteolysis tended (P = 0.09) to be longer for the aspirin compared with the control. This study indicated that oral administration of aspirin may suppress prostaglandin F2α secretion during d 14 to 15 after estrus and may prevent early luteal tissue regression.

HPA axis in major depression: cortisol, clinical symptomatology and genetic variation predict cognition.

The Hypothalamic Pituitary Adrenal (HPA) axis has been implicated in the pathophysiology of a variety of mood and cognitive disorders. Neuroendocrine studies have demonstrated HPA axis overactivity in major depression, a relationship of HPA axis activity to cognitive performance, and a potential role of HPA axis genetic variation in cognition. The present study investigated the simultaneous roles HPA axis activity, clinical symptomatology, and HPA genetic variation play in cognitive performance. Patients with major depression with psychosis (PMD) and without psychosis (NPMD) and healthy controls (HC) were studied. All participants underwent a diagnostic interview and psychiatric ratings, a comprehensive neuropsychological battery, overnight hourly blood sampling for cortisol, and genetic assessment. Cognitive performance differed as a function of depression subtype. Across all subjects, cognitive performance was negatively correlated with higher cortisol, and PMD patients had higher cortisol than did NPMDs and HCs. Cortisol, clinical symptoms, and variation in genes, NR3C1 (glucocorticoid receptor - GR) and NR3C2 (minercorticoid receptor – MR) that encode for glucocorticoid and mineralcorticoid receptors, predicted cognitive performance. Beyond the effects of cortisol, demographics, and clinical symptoms, NR3C1 variation predicted attention and working memory, whereas NR3C2 polymorphisms predicted memory performance. These findings parallel the distribution of GR and MR in primate brain and their putative roles in specific cognitive tasks. HPA axis genetic variation and activity were important predictors of cognition across the entire sample of depressed subjects and healthy controls. GR and MR genetic variation predicted unique cognitive functions, beyond the influence of cortisol and clinical symptoms. GR genetic variation was implicated in attention and working memory, whereas MR was implicated in verbal memory.

Effect of acute interval sprinting exercise on postprandial lipemia of sedentary young men.

[Purpose]Postprandial lipemia (PPL) contributesto the development of atherosclerosis. In females, repeated 8-second bouts of interval sprinting exercise reduced PPL, however, the effect of 8-second bouts of interval sprinting on PPL of overweight males is undetermined. Thus, the effect of 8-secondsof interval sprinting for 20 min, the night before ingestion of a high-fat meal (HFM), on plasma triacylglycerol(TG) levelswas examined.[Methods]Ten overweight males acted as participants (BMI = 26±3.0kg/m2, age 22 ± 2.5 years). A crossover design was employed withinterval sprinting and a noexercise condition separated by 7days. Participants consumed a milkshake (high-fat meal;HFM = 4170 kJ/993 Kcal) the morning after an overnight fast, followed by 4 hourly blood samples. Participants performedone bout of interval sprinting (8seconds sprinting at 110-115rpm, 12seconds active recovery at ~60rpm for 20 minutes) the evening before the consumption of the HFM.[Results]Postprandial TG was 22.5% lower in the interval sprinting compared to the noexercise condition when comparing the change in total area under the curve (ΔAUCT): ISE(7.15±1.90mmolL-1h-1) versus noexercise (9.22±3.44mmolL-1h-1), p=.014. The correlation between fasting TG levels in the noexercise condition and total reduction in AUCT between the conditions was significant (r=.87, p=.001).[Conclusion]One 20-min bout of interval sprinting,the night before consumption of a HFM,significantly attenuated the PPL response of sedentary males.

Fibrinogen concentrate administration inhibits endogenous fibrinogen synthesis in pigs after traumatic hemorrhage.

Fibrinogen plays a central role in coagulation and falls to critical levels early after trauma. Administration of fibrinogen concentrate (FC) to improve hemostasis after severe bleeding seems beneficial, but it is unclear whether its use introduces excessive fibrinogen with a potential risk of thrombosis. This study investigated changes of endogenous fibrinogen metabolism from FC administration following traumatic hemorrhage in pigs. Anesthetized, instrumented pigs were randomized into lactated Ringer's (LR) solution only and LR plus FC groups (n = 7 each). Femur fracture of each pig's left leg was followed by hemorrhage of 60% total blood volume and resuscitation with LR (3× bled volume, LR group) or LR plus FC at 250 mg/kg (LR-FC group). Afterward, a constant infusion of stable isotopes 1-C-phenylalanine (phe, 6 hours) and d5-phe (3 hours) was performed with hourly blood sampling and subsequent gas chromatography-mass spectrometry analysis to quantify fibrinogen synthesis and breakdown rates, respectively. Blood gas and coagulation indices (thromboelastography) were measured on intermittent blood samples, and hemodynamics was continuously monitored. Animals were euthanized after the 6-hour isotope period. Mean arterial pressure decreased by 50% after hemorrhage but improved after LR resuscitation in both groups. Hemorrhage and LR resuscitation reduced total protein, hematocrit, fibrinogen, and platelets to 50% of baseline values. Moreover, hemorrhage and resuscitation decreased fibrinogen concentration (207 ± 6 vs. 132 ± 7 mg/dL) and clot strength (72 ± 2 vs. 63 ± 2 mm) in both groups (p < 0.05). FC administration restored plasma fibrinogen concentrations and clot strength within 15 minutes, while no changes occurred in the LR group. Fibrinogen synthesis rates in the LR-FC group (1.3 ± 0.2 mg/kg/h) decreased versus the LR group (3.1 ± 0.5; p < 0.05), whereas fibrinogen breakdown rates were similar. Our data suggest an effective feedback mechanism that regulates host fibrinogen availability and thereby suggests that acute thrombosis from FC administration is an unlikely risk.

Breaking up of prolonged sitting over three days sustains, but does not enhance, lowering of postprandial plasma glucose and insulin in overweight and obese adults.

To compare the cumulative (3-day) effect of prolonged sitting on metabolic responses during a mixed meal tolerance test (MTT), with sitting that is regularly interrupted with brief bouts of light-intensity walking. Overweight/obese adults (n=19) were recruited for a randomized, 3-day, outpatient, cross-over trial involving: (1) 7-h days of uninterrupted sitting (SIT); and (2) 7-h days of sitting with light-intensity activity breaks [BREAKS; 2-min of treadmill walking (3.2 km/h) every 20 min (total: 17 breaks/day)]. On days 1 and 3, participants underwent a MTT (75 g of carbohydrate, 50 g of fat) and the incremental area under the curve (iAUC) was calculated from hourly blood samples. Generalized estimating equation (GEE) models were adjusted for gender, body mass index (BMI), energy intake, treatment order and pre-prandial values to determine effects of time, condition and time × condition. The glucose iAUC was 1.3 ± 0.5 and 1.5 ± 0.5 mmol·h·l(-1) (mean differences ± S.E.M.) higher in SIT compared with BREAKS on days 1 and 3 respectively (condition effect: P=0.001), with no effect of time (P=0.48) or time × condition (P=0.8). The insulin iAUC was also higher on both days in SIT (day 1: ∆151 ± 73, day 3: ∆91 ± 73 pmol·h·l(-1), P=0.01), with no effect of time (P=0.52) or time × condition (P=0.71). There was no between-treatment difference in triglycerides (triacylglycerols) iAUC. There were significant between-condition effects but no temporal change in metabolic responses to MTT, indicating that breaking up of sitting over 3 days sustains, but does not enhance, the lowering of postprandial glucose and insulin.

Diets rich in starch increase the posthepatic availability of amino acids in dairy cows fed diets at low and normal protein levels

Five mid-lactation multicatheterized Jersey cows were used in a 4×4 Latin square design to investigate whether the increase in milk N yield associated with diets rich in starch versus fiber could originate from changes in the splanchnic AA metabolism and if these changes depended upon the dietary crude protein (CP) content. Four isoenergetic diets were formulated to provide 2 different carbohydrate compositions [diets rich in starch (350g of starch and 310g of neutral detergent fiber/kg of dry matter) versus rich in fiber (45g of starch and 460g of neutral detergent fiber/kg of dry matter)] crossed by 2 different CP contents (12.0 vs. 16.5% CP). At the end of each treatment period, 6 hourly blood samples were collected from the portal and hepatic veins as well as the mesenteric artery to determine net nutrient fluxes across the portal-drained viscera (PDV), liver, and total splanchnic tissues. Dry matter and calculated energy intake as well as total absorbed energy were similar across treatments. However, the net portal appearance (NPA) of acetate, total volatile fatty acids, and β-hydroxybutyrate were higher with diets rich in fiber versus starch, whereas that of oxygen, glucose, butyrate, and insulin were lower. Concomitant to these changes, the percentage of N intake recovered as total AA (TAA) in the portal vein was lower for diets rich in fiber versus starch (42.3 vs. 51.4%, respectively), without, however, any difference observed in the NPA of the main AA used as energy fuels by the PDV (Glu, Gln, and Asp). Despite a higher NPA of TAA with starch versus fiber diets, no differences in the net hepatic flux of TAA, essential and nonessential AA were observed, resulting in a higher (+22%) net splanchnic release of AA and, hence, a greater (+7%) milk N yield. The net hepatic flux and hepatic fractional removal of none of the individual AA was affected as the main carbohydrate changed from fiber to starch, except for Gly and Lys, which were higher for the latter. After correcting for differences in NPA of TAA, the net hepatic uptake of TAA tended to be lower with starch versus fiber diets. The higher transfer of N from feed to milk with diets rich in starch is not the consequence of a direct sparing AA effect of glucogenic diets but rather the result of lower energy requirements by the PDV along with a higher microbial N flow to the duodenum. A better AA use by peripheral tissues with starch versus fiber diets was also hypothesized but more studies are warranted to clarify this issue.