Reverse regulation of soluble receptor for advanced glycation end products and proinflammatory factor resistin and S100A12 in Kawasaki disease

Introduction

Kawasaki disease (KD), an acute febrile disease, characterized by systemic vasculitis, predominantly affects infants and children under 5 years of age. Coronary artery lesions (CALs) are its most critical complication, and the etiology remains unknown yet. In order to explore the value of resistin, S100A12 and soluble receptor for advanced glycation end products (sRAGE) in the pathophysiology of KD, we studied the serum levels of resistin, S100A12 and sRAGE in different stages of KD.

Methods

Serum levels of resistin, S100A12 and sRAGE were measured by enzyme-linked immunosorbent assay (ELISA) method in 15 healthy children and 40 KD patients at acute, afebrile and subacute stage.

Results

The resistin and S100A12 levels, including the ratio of resistin to sRAGE and S100A12 to sRAGE increased significantly in the acute stage, and decreased progressively in the afebrile and subacute stage. However, the sRAGE levels decreased significantly in the acute stage, and increased progressively in the afebrile and subacute stage. In the acute, afebrile and subacute stage, the resistin levels were higher in intravenous immunoglobulin (IVIG) non-responders (0.64 ± 0.30, 0.48 ± 0.35, 0.28 ± 0.19, × 10² ng/ml) than in IVIG responders (0.35 ± 0.24, 0.21 ± 0.19, 0.12 ± 0.05, × 10² ng/ml). In the acute and subacute stage, the S100A12 levels were higher in IVIG non-responders (7.92 ± 2.61, 4.98 ± 4.75, × 10² ng/ml) than in IVIG responders (5.05 ± 3.22, 2.35 ± 2.26, × 10² ng/ml). In the afebrile and subacute stage, the sRAGE levels were lower in IVIG non-responders (3.51 ± 2.64, 3.65 ± 3.27, × 10² pg/ml) than in IVIG responders (6.00 ± 2.78, 7.19 ± 2.88, × 10² pg/ml). The resistin levels were positively correlated with S100A12 levels. The sRAGE levels were negatively related with S100A12 and resistin levels.

Conclusions

Resistin, S100A12 and sRAGE are involved in the pathophysiology of KD.     

Plasma C-Type Natriuretic Peptide as a Predictor for Therapeutic Response to Metoprolol in Children with Postural Tachycardia Syndrome

POTS is a global public-health disease, but predictor for therapeutic response to metoprolol in children with POTS is lacking. This study was designed to investigate predictive value of plasma C-type natriuretic peptide (CNP) in the therapeutic efficacy of metoprolol on postural tachycardia syndrome (POTS) in children. Totally 34 children with POTS and 27 healthy children were included in the study. The head-up test or head-up tilt test was used to check heart rate and blood pressure from supine to upright in subjects. A double antibody (competitive) sandwich immunoluminometric assay was used to detect plasma CNP. Metoprolol was used to treat children with POTS. The difference in plasma concentrations of CNP between responders and non-responders was compared. An ROC curve was used to analyze plasma CNP to predict efficacy of metoprolol on POTS in children. Plasma CNP in children with POTS was significantly higher than that of healthy children [(51.9 ± 31.4) vs. (25.1 ± 19.1) pg/ml, P <0.001]. Plasma CNP in responders to metoprolol was significantly higher than non-responders [(59.1 ± 33.5) vs. (34.8 ± 16.7) pg/ml, P = 0.037] before treatment. The ROC curve showed that area under the curve was 0.821 (95% CI 0.642–0.999). The cut-off value of plasma CNP > 32.55 pg/ml yielded a sensitivity of 95.8% and specificity of 70% in predicting therapeutic efficacy of metoprolol on POTS children. Plasma CNP might serve as a useful predictor for the therapeutic efficacy of metoprolol on POTS in children.     

The Relationship between the Blood Pressure Responses to Exercise following Training and Detraining Periods

Background

Exercise training lowers blood pressure (BP), while BP increases and returns to pre-training values with detraining. Yet, there is considerable variability in these BP responses. We examined the relationship between the BP responses after 6 months of training followed by 2 weeks of detraining among the same people.

Methodology/Principal Findings

Subjects (n = 75) (X+SD, 50.2±10.6 yr) were sedentary, obese, and had prehypertension. They completed an aerobic (n = 34); resistance (n = 28); or aerobic + resistance or concurrent (n = 13) exercise training program. We calculated a metabolic syndrome z score (MetSz). Subjects were classified as BP responders (BP decreased) or non-responders (BP increased) to training and detraining. Linear and multivariable regression tested the BP response. Chi Square tested the frequency of responders and non-responders. The systolic BP (SBP, r = −0.474) and diastolic (DBP, r = −0.540) response to training negatively correlated with detraining (p<0.01), independent of modality (p>0.05). Exercise responders reduced SBP 11.5±7.8 (n = 29) and DBP 9.8±6.2 mmHg (n = 31); non-responders increased SBP 7.9.±10.9 (n = 46) and DBP 4.9±7.1 mmHg (n = 44) (p<0.001). We found 65.5% of SBP training responders were SBP detraining non-responders; while 60.9% of SBP training non-responders were SBP detraining responders (p = 0.034). Similarly, 80.6% of DBP training responders were DBP detraining non-responders; while 59.1% of DBP training non-responders were DBP detraining responders (p<0.001). The SBP detraining response (r = −0.521), resting SBP (r = −0.444), and MetSz (r = 0.288) explained 44.8% of the SBP training response (p<0.001). The DBP detraining response (r = −0.553), resting DBP (r = −0.450), and MetSz (r = 0.463) explained 60.1% of the DBP training response (p<0.001).

Conclusions/Significance

As expected most subjects that decreased BP after exercise training, increased BP after detraining. An unanticipated finding was most subjects that increased BP after exercise training, decreased BP after detraining. Reasons why the negative effects of exercise training on BP maybe reversed with detraining among some people should be explored further.

Trial Registration Information

ClinicalTrials.gov 1R01HL57354; 2003–2008; {"type":"clinical-trial","attrs":{"text":"NCT00275145","term_id":"NCT00275145"}}NCT00275145     

PRE-EXERCISE ARGININE SUPPLEMENTATION INCREASES TIME TO EXHAUSTION IN ELITE MALE WRESTLERS

Dietary supplements containing arginine are among the most popular ergogenics intended to enhance strength, power and muscle recovery associated with both anaerobic and aerobic exercise. The aim of the present study was to evaluate the possible effect of pre-exercise acute intake of arginine on performance and exercise metabolism during incremental exhaustive exercise in elite male wrestlers. Nine volunteer elite male wrestlers (24.7±3.8 years) participated in this study. The test-retest protocol was used on the same subjects. The study was conducted using a cross-over design. A single dose of arginine (1.5 g · 10 kg^-1 body weight) or placebo was given to the subjects after 12 hours fasting (during the night) for both test and retest. Subjects were allowed to drink water but not allowed to eat anything between arginine or placebo ingestion and the exercise protocol. An incremental exercise protocol was applied and oxygen consumption was measured during the exercise. Heart rate and plasma lactate levels were measured during the exercise and recovery. Results showed that in the same working loads there was no significant difference for the mean lactate levels and no difference in maximum oxygen consumption (arginine 52.47±4.01 mL · kg^-1 · min^-1, placebo 52.07±5.21 mL · kg^-1 · min^-1) or in maximum heart rates (arginine 181.09±13.57 bpm, placebo 185.89±7.38 bpm) between arginine and placebo trials. Time to exhaustion was longer with arginine supplementation (1386.8±69.8 s) compared to placebo (1313±90.8 s) (p < 0.05). These results suggest that L-arginine supplementation can have beneficial effects on exercise performance in elite male wrestlers but cannot explain the metabolic pathways which are responsible from these effects.     

Improvement of Left Ventricular Function under Cardiac Resynchronization Therapy Goes along with a Reduced Incidence of Ventricular Arrhythmia

Objectives

The beneficial effects of cardiac resynchronization therapy (CRT) are thought to result from favorable left ventricular (LV) reverse remodeling, however CRT is only successful in about 70% of patients. Whether response to CRT is associated with a decrease in ventricular arrhythmias (VA) is still discussed controversially. Therefore, we investigated the incidence of VA in CRT responders in comparison with non-responders.

Methods

In this nonrandomized, two-center, observational study patients with moderate-to-severe heart failure, LV ejection fraction (LVEF) ≤35%, and QRS duration >120 ms undergoing CRT were included. After 6 months patients were classified as CRT responders or non-responders. Incidence of VA was compared between both groups by Kaplan-Meier analysis and Cox regression analysis. ROC analysis was performed to determine the aptitude of LVEF cut-off values to predict VA.

Results

In total 126 consecutive patients (64±11years; 67%male) were included, 74 were classified as responders and 52 as non-responders. While the mean LVEF at baseline was comparable in both groups (25±7% vs. 24±8%; P = 0.4583) only the responder group showed an improvement of LVEF (36±6% vs. 24±7; p<0.0001) under CRT. In total in 56 patients VA were observed during a mean follow-up of 28±14 months, with CRT responders experiencing fewer VA than non-responders (35% vs. 58%, p<0.0061). Secondary preventive CRT implantation was associated with a higher likelihood of VA. As determined by ROC analysis an increase of LVEF by >7% was found to be a predictor of a significantly lower incidence of VA (AUC = 0.606).

Conclusions

Improvement of left ventricular function under cardiac resynchronization therapy goes along with a reduced incidence of ventricular arrhythmia.     

Determination of base and backbone contributions to the thermodynamics of premelting and melting transitions in B DNA

In previous papers of this series the temperature-dependent Raman spectra of poly(dA)·poly(dT) and poly(dA–dT)·poly(dA–dT) were used to characterize structurally the melting and premelting transitions in DNAs containing consecutive A·T and alternating A·T/T·A base pairs. Here, we describe procedures for obtaining thermodynamic parameters from the Raman data. The method exploits base-specific and backbone-specific Raman markers to determine separate thermodynamic contributions of A, T and deoxyribosyl-phosphate moieties to premelting and melting transitions. Key findings include the following: (i) Both poly(dA)·poly(dT) and poly(dA–dT)· poly(dA–dT) exhibit robust premelting transitions, due predominantly to backbone conformational changes. (ii) The significant van’t Hoff premelting enthalpies of poly(dA)·poly(dT) [ΔH_vH^pm = 18.0 ± 1.6 kcal·mol^–1 (kilocalories per mole cooperative unit)] and poly(dA–dT)·poly(dA–dT) (ΔH_vH^pm = 13.4 ± 2.5 kcal·mol^–1) differ by an amount (~4.6 kcal·mol^–1) estimated as the contribution from three-centered inter-base hydrogen bonding in (dA)_n·(dT)_n tracts. (iii) The overall stacking free energy of poly(dA)· poly(dT) [–6.88 kcal·mol_bp^–1 (kilocalories per mole base pair)] is greater than that of poly(dA–dT)· poly(dA–dT) (–6.31 kcal·mol_bp^–1). (iv) The difference between stacking free energies of A and T is significant in poly(dA)·poly(dT) (ΔΔG_st = 0.8 ± 0.3 kcal· mol_bp^–1), but marginal in poly(dA–dT)·poly(dA–dT) (ΔΔG_st = 0.3 ± 0.3 kcal·mol_bp^–1). (v) In poly(dA)· poly(dT), the van’t Hoff parameters for melting of A (ΔH_vH^A = 407 ± 23 kcal·mol^–1, ΔS_vH^A = 1166 ± 67 cal·°K^–1·mol^–1, ΔG_vH(25°C)^A = 60.0 ± 3.2 kcal·mol^–1) are clearly distinguished from those of T (ΔH_vH^T = 185 ± 38 kcal·mol^–1, ΔS_vH^T = 516 ± 109 cal·°K^–1·mol^–1, ΔG_vH(25°C)^T = 27.1 ± 5.5 kcal·mol^–1). (vi) Similar relative differences are observed in poly(dA–dT)· poly(dA–dT) (ΔH_vH^A = 333 ± 54 kcal·mol^–1, ΔS_vH^A = 961 ± 157 cal·°K^–1·mol^–1, ΔG_vH(25°C)^A = 45.0 ± 7.6 kcal· mol^–1; ΔH_vH^T = 213 ± 30 kcal·mol^–1, ΔS_vH^T = 617 ± 86 cal·°K^–1·mol^–1, ΔG_vH(25°C)^T = 29.3 ± 4.9 kcal·mol^–1). The methodology employed here distinguishes thermodynamic contributions of base stacking, base pairing and backbone conformational ordering in the molecular mechanism of double-helical B DNA formation.     

Can Positron Emission Tomography/Computed Tomography with the Dual Tracers Fluorine-18 Fluoroestradiol and Fluorodeoxyglucose Predict Neoadjuvant Chemotherapy Response of Breast Cancer? ----A Pilot Study

Objective

To assess the clinical value of dual tracers Positron emission tomography/computed tomography (PET/CT) ¹⁸F-fluoroestradiol (¹⁸F-FES) and ¹⁸F-fluorodeoxyglucose (¹⁸F-FDG) in predicting neoadjuvant chemotherapy response (NAC) of breast cancer.

Methods

Eighteen consecutive patients with newly diagnosed, non-inflammatory, stage II and III breast cancer undergoing NAC were included. Before chemotherapy, they underwent both ¹⁸F-FES and ¹⁸F-FDG PET/CT scans. Surgery was performed after three to six cycles of chemotherapy. Tumor response was graded and divided into two groups: the responders and non-responders. We used the maximum standardized uptake value (SUVmax) to qualify each primary lesion.

Results

Pathologic analysis revealed 10 patients were responders while the other 8 patients were non-responders. There was no statistical difference of SUVmax-FDG and tumor size between these two groups (P>0.05). On the contrary, SUVmax-FES was lower in responders (1.75±0.66 versus 4.42±1.14; U=5, P=0.002); and SUVmax-FES/FDG also showed great value in predicting outcome (0.16±0.06 versus 0.54±0.22; U=5, P=0.002).

Conclusions

Our study showed ¹⁸F-FES PET/CT might be feasible to predict response of NAC. However, whether the use of dual tracers ¹⁸F-FES and ¹⁸F-FDG has complementary value should be further studied.     

Evaluation of novel SexedULTRA-4M technology for in vitro bovine embryo production

SexedULTRA-4M™ is made using an improved method of sex-sorting sperm in a less damaging environment for better retaining sperm integrity throughout the sorting process. The objective of this research was to compare conventional (CONV) and SexedULTRA-4M™ (ULTRA-4M) semen for bovine IVP using four Angus bulls. Matured slaughterhouse oocytes (n = 4000) were divided into the CONV group and the ULTRA-4M group (2000 COCs for each semen type). The IVF process was implemented with CONV and ULTRA-4M semen from the same bull. The cleavage rates, eight cell embryos and blastocysts on day 7 of culture were evaluated for each semen type and each bull. The statistical analysis was carried out with the ANOVA procedure SAS software. The results were 54.45% ± 1.03 and 58.10% ± 1.07; 35% ± 1.57 and 39.15% ± 1.62; 22.8% ± 1.09 and 27.15% ± 1.12 for CONV and ULTRA-4M, respectively, for cleavage rate, eight cell embryos and blastocysts on day 7 for the average of all bulls, comparing only the semen type. Concerning only the semen type, ULTRA-4M was significantly superior to CONV for cleavage rates (P = 0.01) and blastocysts on day 7 (P = 0.009). There were no significant differences between the CONV and ULTRA-4M groups (P>0.05) for all variables analyzed for Bull 1 and Bull 4, however, for Bull 2 ULTRA-4M was significantly superior to CONV for cleavage rates and blastocysts on day 7 (P< 0.05). In Bull 3, ULTRA-4M was significantly higher (P< 0.05) for blastocysts on day 7 compared to CONV. In conclusion, under the conditions of this research the ULTRA-4M and CONV semen produced similar bovine IVP results overall.     

Mass Change of Glaciers in Muztag Ata–Kongur Tagh,Eastern Pamir,China from 1971/76 to 2013/14 as Derived from Remote Sensing Data

The assessment of glacier mass budget is crucial for assessing water reserves stored in glaciers. Derived glacier mass changes in the Muztag Ata and Kongur Tagh (MAKT) region in the eastern Pamir, northwestern China, is helpful in improving our knowledge of the dynamics of glaciers under a changing climate in High Mountain Asia. Here, glacier area and mass changes derived from remote sensing data are investigated for the period 1971/76–2013/14 for glaciers in MAKT. We have used ASTER images (2013/14), Cartosat-1 (2014) and Landsat, SRTM (Shuttle Radar Terrain Mission) digital elevation model (DEM) (2000), topographic maps (1971/76) and the first and second Chinese glacier inventories (CGIs). Our results indicated that the glacier area of MAKT decreased from 1018.3 ± 12.99 km² in 1971/76 to 999.2 ± 31.22 km² in 2014 (–1.9 ± 0.2%). Weak area shrinkage of glaciers by 2.5 ± 0.5 km² (0.2 ± 0.1%) happened after 2000 and the period 2009–2014 even saw a slight expansion by 0.5 ± 0.1 km² (0.1 ± 0.0%). The glaciers in this region have experienced an overall loss of –6.99 ± 0.80 km³ in ice volume or –0.15 ± 0.12 m water equivalent (w.e.) a^–1 from 1971/76 to 2013/14. The mass budget of MAKT was –0.19 ± 0.19 m w.e. a⁻¹ for the period ~1971/76–1999 and –0.14 ± 0.24 m w.e. a⁻¹ during 1999–2013/2014. Similar to previous studies, there has been little mass change in the Pamir over recent decades despite such uncertainties. Glacier mass change showed spatial and temporal heterogeneity, with strong mass loss on debris-covered glaciers with an average of –0.32 ± 0.12 m w.e. a⁻¹ from the 1970s to 2013/14.     

Microzooplankton Growth Rates Examined across a Temperature Gradient in the Barents Sea

Growth rates (µ) of abundant microzooplankton species were examined in field experiments conducted at ambient sea temperatures (−1.8–9.0°C) in the Barents Sea and adjacent waters (70–78.5°N). The maximum species-specific µ of ciliates and athecate dinoflagellates (0.33–1.67 d⁻¹ and 0.52–1.14 d⁻¹, respectively) occurred at temperatures below 5°C and exceeded the µ_max predicted by previously published, laboratory culture-derived equations. The opposite trend was found for thecate dinoflagellates, which grew faster in the warmer Atlantic Ocean water. Mixotrophic ciliates and dinoflagellates grew faster than their heterotrophic counterparts. At sub-zero temperatures, microzooplankton µ_max matched those predicted for phytoplankton by temperature-dependent growth equations. These results indicate that microzooplankton protists may be as adapted to extreme Arctic conditions as their algal prey.     

Validity of Cooper’s 12-minute run test for estimation of maximum oxygen uptake in male university students

The present study was conducted to validate the applicability of Cooper''s 12-minute run test (CRT) for predicting VO₂max in male university students of Kolkata, India, to bypass the exhaustive and complicated protocol of direct estimation of VO₂max. Eighty-eight sedentary male university students recruited by simple random sampling from the University of Calcutta, Kolkata, were randomly assigned to the study group (N = 58) and the confirmatory group (N = 30). VO₂max of each participant was determined by the direct procedure and the indirect CRT method. The mean value of predicted VO₂max (PVO₂max) (42.8±4.0 ml · kg⁻¹ · min⁻¹ with a range of 33.7–50.9) showed a significant difference with VO₂max (39.8±4.0 ml · kg⁻¹ · min⁻¹ with a range of 33.5–47.7) in the study group. Limits of agreement between PVO₂max and VO₂max were large enough (0.10 to 5.94 ml · kg⁻¹ · min⁻¹) with poor confidence intervals indicating inapplicability of the current protocol of CRT in the studied population. The prediction norm [Y = 21.01X – 11.04 (SEE = 0.193 ml · kg⁻¹ · min⁻¹)] was computed from the significant correlation (r = 0.93, P < 0.001) between distance covered in CRT and VO₂max. Application of this norm in the confirmatory group revealed an insignificant difference between PVO₂max and VO₂max. The modified equation is recommended for application of CRT as a valid method to evaluate the cardiorespiratory fitness in terms of VO₂max in sedentary male Indian youth.     

Whole-Body Insulin Sensitivity Rather than Body-Mass-Index Determines Fasting and Post-Glucose-Load Growth Hormone Concentrations

Background

Obese, non-acromegalic persons show lower growth hormone (GH) concentrations at fasting and reduced GH nadir during an oral glucose tolerance test (OGTT). However, this finding has never been studied with regard to whole-body insulin-sensitivity as a possible regulator.

Methods

In this retrospective analysis, non-acromegalic (NonACRO, n = 161) and acromegalic (ACRO, n = 35), non-diabetic subjects were subdivided into insulin-sensitive (IS) and –resistant (IR) groups according to the Clamp-like Index (CLIX)-threshold of 5 mg·kg⁻¹·min⁻¹ from the OGTT.

Results

Non-acromegalic IS (CLIX: 8.8±0.4 mg·kg⁻¹·min⁻¹) persons with similar age and sex distribution, but lower (p<0.001) body-mass-index (BMI = 25±0 kg/m², 84% females, 56±1 years) had 59% and 70%, respectively, higher (p<0.03) fasting GH and OGTT GH area under the curve concentrations than IR (CLIX: 3.5±0.1 mg·kg⁻¹·min⁻¹, p<0.001) subjects (BMI = 29±1 kg/m², 73% females, 58±1 years). When comparing on average overweight non-acromegalic IS and IR with similar anthropometry (IS: BMI: 27±0 kg/m², 82% females, 58±2 years; IR: BMI: 27±0 kg/m², 71% females, 60±1 years), but different CLIX (IS: 8.7±0.9 vs. IR: 3.8±0.1 mg·kg⁻¹·min⁻¹, p<0.001), the results remained almost the same. In addition, when adjusted for OGTT-mediated glucose rise, GH fall was less pronounced in IR. In contrast, in acromegalic subjects, no difference was found between IS and IR patients with regard to fasting and post-glucose-load GH concentrations.

Conclusions

Circulating GH concentrations at fasting and during the OGTT are lower in non-acromegalic insulin-resistant subjects. This study seems the first to demonstrate that insulin sensitivity rather than body-mass modulates fasting and post-glucose-load GH concentrations in non-diabetic non–acromegalic subjects.     

Decadal Changes in the World's Coastal Latitudinal Temperature Gradients

Most of the world''s living marine resources inhabit coastal environments, where average thermal conditions change predictably with latitude. These coastal latitudinal temperature gradients (CLTG) coincide with important ecological clines,e.g., in marine species diversity or adaptive genetic variations, but how tightly thermal and ecological gradients are linked remains unclear. A first step is to consistently characterize the world''s CLTGs. We extracted coastal cells from a global 1°×1° dataset of weekly sea surface temperatures (SST, 1982–2012) to quantify spatial and temporal variability of the world''s 11 major CLTGs. Gradient strength, i.e., the slope of the linear mean-SST/latitude relationship, varied 3-fold between the steepest (North-American Atlantic and Asian Pacific gradients: −0.91°C and −0.68°C lat⁻¹, respectively) and weakest CLTGs (African Indian Ocean and the South- and North-American Pacific gradients: −0.28, −0.29, −0.32°C lat⁻¹, respectively). Analyzing CLTG strength by year revealed that seven gradients have weakened by 3–10% over the past three decades due to increased warming at high compared to low latitudes. Almost the entire South-American Pacific gradient (6–47°S), however, has considerably cooled over the study period (−0.3 to −1.7°C, 31 years), and the substantial weakening of the North-American Atlantic gradient (−10%) was due to warming at high latitudes (42–60°N, +0.8 to +1.6°C,31 years) and significant mid-latitude cooling (Florida to Cape Hatteras 26–35°N, −0.5 to −2.2°C, 31 years). Average SST trends rarely resulted from uniform shifts throughout the year; instead individual seasonal warming or cooling patterns elicited the observed changes in annual means. This is consistent with our finding of increased seasonality (i.e., summer-winter SST amplitude) in three quarters of all coastal cells (331 of 433). Our study highlights the regionally variable footprint of global climate change, while emphasizing ecological implications of changing CLTGs, which are likely driving observed spatial and temporal clines in coastal marine life.     

Carriers of Loss-of-Function Mutations in EXT Display Impaired Pancreatic Beta-Cell Reserve Due to Smaller Pancreas Volume

Exotosin (EXT) proteins are involved in the chain elongation step of heparan sulfate (HS) biosynthesis, which is intricately involved in organ development. Loss of function mutations (LOF) in EXT1 and EXT2 result in hereditary exostoses (HME). Interestingly, HS plays a role in pancreas development and beta-cell function, and genetic variations in EXT2 are associated with an increased risk for type 2 diabetes mellitus. We hypothesized that loss of function of EXT1 or EXT2 in subjects with hereditary multiple exostoses (HME) affects pancreatic insulin secretion capacity and development. We performed an oral glucose tolerance test (OGTT) followed by hyperglycemic clamps to investigate first-phase glucose-stimulated insulin secretion (GSIS) in HME patients and age and gender matched non-affected relatives. Pancreas volume was assessed with magnetic resonance imaging (MRI). OGTT did not reveal significant differences in glucose disposal, but there was a markedly lower GSIS in HME subjects during hyperglycemic clamp (iAUC HME: 0.72 [0.46–1.16] vs. controls 1.53 [0.69–3.36] nmol·l⁻¹·min⁻¹, p<0.05). Maximal insulin response following arginine challenge was also significantly attenuated (iAUC HME: 7.14 [4.22–10.5] vs. controls 10.2 [7.91–12.70] nmol·l⁻¹·min⁻¹ p<0.05), indicative of an impaired beta-cell reserve. MRI revealed a significantly smaller pancreatic volume in HME subjects (HME: 72.0±15.8 vs. controls 96.5±26.0 cm³ p = 0.04). In conclusion, loss of function of EXT proteins may affect beta-cell mass and insulin secretion capacity in humans, and render subjects at a higher risk of developing type 2 diabetes when exposed to environmental risk factors.     

Hydropathic analysis of the free energy differences in anthracycline antibiotic binding to DNA

Molecular models of six anthracycline antibiotics and their complexes with 32 distinct DNA octamer sequences were created and analyzed using HINT (Hydropathic INTeractions) to describe binding. The averaged binding scores were then used to calculate the free energies of binding for comparison with experimentally determined values. In parsing our results based on specific functional groups of doxorubicin, our calculations predict a free energy contribution of –3.6 ± 1.1 kcal mol^–1 (experimental –2.5 ± 0.5 kcal mol^–1) from the groove binding daunosamine sugar. The net energetic contribution of removing the hydroxyl at position C9 is –0.7 ± 0.7 kcal mol^–1 (–1.1 ± 0.5 kcal mol^–1). The energetic contribution of the 3′ amino group in the daunosamine sugar (when replaced with a hydroxyl group) is –3.7 ± 1.1 kcal mol^–1 (–0.7 ± 0.5 kcal mol^–1). We propose that this large discrepancy may be due to uncertainty in the exact protonation state of the amine. The energetic contribution of the hydroxyl group at C14 is +0.4 ± 0.6 kcal mol^–1 (–0.9 ± 0.5 kcal mol^–1), largely due to unfavorable hydrophobic interactions between the hydroxyl oxygen and the methylene groups of the phosphate backbone of the DNA. Also, there appears to be considerable conformational uncertainty in this region. This computational procedure calibrates our methodology for future analyses where experimental data are unavailable.     

Pulse Wave Transit Time Measurements of Cardiac Output in Septic Shock Patients: A Comparison of the Estimated Continuous Cardiac Output System with Transthoracic Echocardiography

Background

We determined reliability of cardiac output (CO) measured by pulse wave transit time cardiac output system (esCCO system; CO_esCCO) vs transthoracic echocardiography (CO_TTE) in mechanically ventilated patients in the early phase of septic shock. A secondary objective was to assess ability of esCCO to detect change in CO after fluid infusion.

Methods

Mechanically ventilated patients admitted to the ICU, aged >18 years, in sinus rhythm, in the early phase of septic shock were prospectively included. We performed fluid infusion of 500ml of crystalloid solution over 20 minutes and recorded CO by EsCCO and TTE immediately before (T0) and 5 minutes after (T1) fluid administration. Patients were divided into 2 groups (responders and non-responders) according to a threshold of 15% increase in CO_TTE in response to volume expansion.

Results

In total, 25 patients were included, average 64±15 years, 15 (60%) were men. Average SAPSII and SOFA scores were 55±21.3 and 13±2, respectively. ICU mortality was 36%. Mean cardiac output at T0 was 5.8±1.35 L/min by esCCO and 5.27±1.17 L/min by CO_TTE. At T1, respective values were 6.63 ± 1.57 L/min for esCCO and 6.10±1.29 L/min for CO_TTE. Overall, 12 patients were classified as responders, 13 as non-responders by the reference method. A threshold of 11% increase in CO_esCCO was found to discriminate responders from non-responders with a sensitivity of 83% (95% CI, 0.52-0.98) and a specificity of 77% (95% CI, 0.46-0.95).

Conclusion

We show strong correlation esCCO and echocardiography for measuring CO, and change in CO after fluid infusion in ICU patients.     

Rates of entry and oxidation of acetate,glucose, d(?)-β-hydroxybutyrate,palmitate, oleate and stearate,and rates of production and oxidation of propionate and butyrate in fed and starved sheep

1. Rates of entry and oxidation of a range of metabolites have been measured in tracheostomized sheep (diet, 800g. of lucerne chaff and 100g. of maize/day) by combining isotope-dilution techniques with the continuous measurement of total respiratory gas exchange, and ¹⁴CO₂ production during the intravenous or intraruminal infusion of ¹⁴C-labelled substrates. 2. Mean entry rates in fed and starved (24hr.) sheep respectively, expressed as mg./min./kg. body wt.^0·75, were: glucose, 5·0 (range 4·8–5·1, 2 observations) and 3·8 (3·2–4·2, 4); acetate, 10·8 (9·1–13·5, 4) and 5·8 (1); d(−)-β-hydroxybutyrate, 1·4 (1) and 1·5 (0·8–2·4, 4); palmitate, oleate and stearate (starved sheep only) 1·0 (0·6–1·9, 7), 0·9 (0·2–1·6, 10) and 0·9 (0·5–1·1, 11) respectively. 3. Production rates of propionate and butyrate in continuously feeding sheep were 6·4 (4·7–8·3, 4) and 4·3 (3·4–6·1, 4) mg./min./kg.^0·75 respectively, and in starved (24hr.) sheep were 2·5 (2·2–2·9, 2) and 1·0 (0·8–1·2, 2) mg./min./kg.^0·75 respectively. 4. Calculated terminal values for the specific radioactivity of respiratory ¹⁴CO₂ during measurements of entry rates and production rates were used to calculate the contributions of individual substrates to overall oxidative metabolism. Mean values for fed and starved sheep respectively were: glucose, 9·1 (8·6–9·6, 2) and 11·2 (5·9–15·1, 4)%; acetate, 31·6 (26·8–38·1, 4) and 22·1 (1)%; d(−)-β-hydroxybutyrate, 10·4 (1) and 4·8 (1·9–7·7, 4)%; propionate, 23·0 (13·8–29·9, 4) and 7·1 (6·8–7·4, 2)%; butyrate, 16·5 (13·7–20·5, 4) and 5·3 (5·2–5·3, 2)%; palmitate, oleate and stearate (starved sheep only), 4·7 (2·0–7·7, 7), 4·0 (1·2–6·6, 10) and 4·4 (3·8–5·8, 9)% respectively. The sum of these values for individual substrates in fed and starved sheep, excluding that of β-hydroxybutyrate and after correction of the glucose value for the known interrelations of this substrate with propionate, accounted for 76% and 58% respectively of total production of carbon dioxide. 5. Calculations based on the proportion of substrate entry directly oxidized indicated that the substrates studied accounted for 63% (fed sheep) and 43% (starved sheep) of total energy expenditure measured by oxygen uptake. The contribution of β-hydroxybutyrate was excluded, and corrections were made for glucose–propionate interrelations, and for the different rates of oxidation of the methyl and carboxyl fragments of acetate. 6. The present results have been combined with those obtained earlier in this Laboratory to examine the relationships between rates of substrate entry and oxidation, and concentrations of substrate in blood. Rates of entry of acetate, glucose, d(−)-β-hydroxybutyrate, palmitate and oleate (but not stearate) were well correlated with concentration in blood, and substrate contribution to production of carbon dioxide showed a similar correlation to blood concentration, except with glucose. 7. It was concluded that the general technique is of potential value in providing valid quantitative parameters of animal metabolism.     

Global trends and uncertainties in terrestrial denitrification and N2O emissions

Soil nitrogen (N) budgets are used in a global, distributed flow-path model with 0.5° × 0.5° resolution, representing denitrification and N₂O emissions from soils, groundwater and riparian zones for the period 1900–2000 and scenarios for the period 2000–2050 based on the Millennium Ecosystem Assessment. Total agricultural and natural N inputs from N fertilizers, animal manure, biological N₂ fixation and atmospheric N deposition increased from 155 to 345 Tg N yr⁻¹ (Tg = teragram; 1 Tg = 10¹² g) between 1900 and 2000. Depending on the scenario, inputs are estimated to further increase to 408–510 Tg N yr⁻¹ by 2050. In the period 1900–2000, the soil N budget surplus (inputs minus withdrawal by plants) increased from 118 to 202 Tg yr⁻¹, and this may remain stable or further increase to 275 Tg yr⁻¹ by 2050, depending on the scenario. N₂ production from denitrification increased from 52 to 96 Tg yr⁻¹ between 1900 and 2000, and N₂O–N emissions from 10 to 12 Tg N yr⁻¹. The scenarios foresee a further increase to 142 Tg N₂–N and 16 Tg N₂O–N yr⁻¹ by 2050. Our results indicate that riparian buffer zones are an important source of N₂O contributing an estimated 0.9 Tg N₂O–N yr⁻¹ in 2000. Soils are key sites for denitrification and are much more important than groundwater and riparian zones in controlling the N flow to rivers and the oceans.     

The characterization and localization of frog skin cholinesterase

Frog skin cholinesterase is largely of the serum (pseudocholinesterase) type. For whole skin, the activity at 10^–1 AcChCl is 4.9 µl./mg. N/hr. Tela subcutanea isolated by dissection exhibits an activity of 65 µl./mg. N/hr. at 10^–1 AcChCl. Since about one-tenth of the nitrogen of the skin is located in the tela subcutanea, it is estimated that more than 90 per cent of the enzyme is associated with this tissue layer.     

Movement Demands of Elite Under-20s and Senior International Rugby Union Players

This study compared the movement demands of elite international Under-20 age grade (U20s) and senior international rugby union players during competitive tournament match play. Forty elite professional players from an U20 and 27 elite professional senior players from international performance squads were monitored using 10Hz global positioning systems (GPS) during 15 (U20s) and 8 (senior) international tournament matches during the 2014 and 2015 seasons. Data on distances, velocities, accelerations, decelerations, high metabolic load (HML) distance and efforts, and number of sprints were derived. Data files from players who played over 60 min (n = 258) were separated firstly into Forwards and Backs, and more specifically into six positional groups; FR–Front Row (prop & hooker), SR–Second Row, BR–Back Row (Flankers & No.8), HB–Half Backs (scrum half & outside half), MF–Midfield (centres), B3 –Back Three (wings & full back) for match analysis. Linear mixed models revealed significant differences between U20 and senior teams in both the forwards and backs. In the forwards the seniors covered greater HML distance (736.4 ± 280.3 vs 701.3 ± 198.7m, p = 0.01) and severe decelerations (2.38 ± 2.2 vs 2.28 ± 1.65, p = 0.05) compared to the U20s, but performed less relative HSR (3.1 ± 1.6 vs 3.2 ± 1.5, p < 0.01), moderate (19.4 ± 10.5 vs 23.6 ± 10.5, p = 0.01) and high accelerations (2.2 ± 1.9 vs 4.3 ± 2.7, p < 0.01) and sprint•min^-1 (0.11 ± 0.06 vs 0.11 ± 0.05, p < 0.01). Senior backs covered a greater relative distance (73.3 ± 8.1 vs 69.1 ± 7.6 m•min^-1, p < 0.01), greater High Metabolic Load (HML) distance (1138.0 ± 233.5 vs 1060.4 ± 218.1m, p < 0.01), HML efforts (112.7 ± 22.2 vs 98.8 ± 21.7, p < 0.01) and heavy decelerations (9.9 ± 4.3 vs 9.5 ± 4.4, p = 0.04) than the U20s backs. However, the U20s backs performed more relative HSR (7.3 ± 2.1 vs 7.2 ± 2.1, p <0.01) and sprint•min^-1 (0.26 ± 0.07 vs 0.25 ± 0.07, p < 0.01). Further investigation highlighted differences between the 6 positional groups of the teams. The positional groups that differed the most on the variables measured were the FR and MF groups, with the U20s FR having higher outputs on HSR, moderate & high accelerations, moderate, high & severe decelerations, HML distance, HML efforts, and sprints•min^-1. For the MF group the senior players produced greater values for relative distance covered, HSR, moderate decelerations, HML distance and sprint•min^-1. The BR position group was most similar with the only differences seen on heavy accelerations (U20s higher) and moderate decelerations (seniors higher). Findings demonstrate that U20s internationals appear to be an adequate ‘stepping stone’ for preparing players for movement characteristics found senior International rugby, however, the current study highlight for the first time that certain positional groups may require more time to be able to match the movement demands required at a higher playing level than others. Conditioning staff must also bear in mind that the U20s players whilst maintaining or improving match movement capabilities may require to gain substantial mass in some positions to match their senior counterparts.