Acute supra-therapeutic oral terbutaline administration has no ergogenic effect in non-asthmatic athletes.

This study aimed to investigate the effects on a possible improvement in aerobic and anaerobic performance of oral terbutaline (TER) at a supra-therapeutic dose in 7 healthy competitive male athletes. On day 1, ventilatory threshold, maximum oxygen uptake [Formula: see text] and corresponding power output were measured and used to determine the exercise load on days 2 and 3. On days 2 and 3, 8 mg of TER or placebo were orally administered in a double-blind process to athletes who rested for 3 h, and then performed a battery of tests including a force-velocity exercise test, running sprint and a maximal endurance cycling test at Δ50 % (50 % between VT and [Formula: see text]). Lactatemia, anaerobic parameters and endurance performance ([Formula: see text] and time until exhaustion) were raised during the corresponding tests. We found that TER administration did not improve any of the parameters of aerobic performance (p > 0.05). In addition, no change in [Formula: see text] kinetic parameters was found with TER compared to placebo (p > 0.05). Moreover, no enhancement of the force-velocity relationship was observed during sprint exercises after TER intake (p > 0.05) and, on the contrary, maximal strength decreased significantly after TER intake (p < 0.05) but maximal power remained unchanged (p > 0.05). In conclusion, oral acute administration of TER at a supra-therapeutic dose seems to be without any relevant ergogenic effect on anaerobic and aerobic performances in healthy athletes. However, all participants experienced adverse side effects such as tremors.

Skeletal muscle mitochondria in the elderly: effects of physical fitness and exercise training.

Context: Sarcopenia is thought to be associated with mitochondrial (M) loss. It is unclear whether the decrease in M content is consequent to aging per se or to decreased physical activity. Objectives: To examine the influence of fitness on M content and function, and to assess whether exercise could improve M function in older adults. Design and subjects: Three distinct studies were conducted: 1) a cross-sectional observation comparing M content and fitness in a large heterogeneous cohort of older adults; 2) a case-control study comparing chronically endurance-trained older adults (A) and sedentary (S) subjects matched for age and gender; 3) a 4-month exercise intervention in S. Setting: University-based clinical research center Outcomes: M volume density (Mv) was assessed by electron microscopy from vastus lateralis biopsies, electron transport chain proteins (ETC) by western blotting, mRNAs for transcription factors involved in M biogenesis by qRT-PCR and in-vivo oxidative capacity (ATPmax) by (31)P-MR spectroscopy. Peak oxygen uptake (VO2peak) was measured by GXT. Results: VO2peak was strongly correlated with Mv in eighty 60-80 yo adults. Comparison of A vs. S revealed differences in Mv, ATPmax and some ETC complexes. Finally, exercise intervention confirmed that S are able to recover Mv, ATPmax and specific transcription factors. Conclusions: These data suggest that 1) aging per se is not the primary culprit leading to M dysfunction, 2) an aerobic exercise program, even at an older age, can ameliorate the loss in skeletal muscle M content and may prevent aging muscle comorbidities and 3) the improvement of M function is all about content.

Transport of a biocontrol Pseudomonas fluorescens through 2.5-m deep outdoor lysimeters and survival in the effluent water

Application of wild-type or genetically-modified bacteria to the soil environment entails the risk of dissemination of these organisms to the groundwater. To measure vertical transport of bacteria under natural climatic conditions, Pseudomonas fluorescens strain CHA0 was released together with bromide as a mobile tracer at the surface of large outdoor lysimeters. Two experiments, one starting in autumn 1993 and the other in spring 1994 were performed. Shortly after a heavy rainfall in late spring 1994, the released bacteria were detected for the first time in effluent water from the 2.5-m-deep lysimeters in both experiments, i.e. 210 d and 21 d, respectively, after inoculation. Only a 10−9 to 10−8 fraction of the inoculum was recovered as culturable cells in the effluent water, but a larger fraction of the CHA0 cells was in a non-culturable state as detected with immunofluorescence microscopy. As much as 50% of the mobile tracer percolated through the lysimeters, indicating that, compared with bromide, bacterial cells were retained in soil. In the second part of this study, persistence of CHA0 in groundwater microcosms consisting of lysimeter effluent water was studied for 380 d. Survival of the inoculant as culturable cells was better under anaerobic than under aerobic conditions. However, a large fraction of the cells became non-culturable in both cases. When the experiment was performed with filter-sterilized effluent water, the total count of introduced bacteria did not decline with time. In conclusion, the biocontrol strain was transported in low numbers to a potential groundwater level under natural climatic conditions, but could persist for an extended period in groundwater microcosms.

Microbial alterations of the soil influenced by induced compaction

Compaction is one of the most destructive factors of soil quality, however the effects on the microbial community and enzyme activity have not been investigated in detail so far. The objective of this study was to evaluate the effects of soil compaction caused by the traffic of agricultural machines on the soil microbial community and its enzyme activity. Six compaction levels were induced by tractors with different weights driving over a Eutrustox soil and the final density was measured. Soil samples were collected after corn from the layers 0-0.10 and 0.10-0.20 m. The compaction effect on all studied properties was evident. Total bacteria counts were reduced significantly (by 22-30 %) and by 38-41 % of nitrifying bacteria in the soil with highest bulk density compared to the control. On the other hand, fungi populations increased 55-86 % and denitrifying bacteria 49-53 %. Dehydrogenase activity decreased 20-34 %, urease 44-46 % and phosphatase 26-28 %. The organic matter content and soil pH decreased more in the 0-0.10 than in the 0.10-0.20 m layer and possibly influenced the reduction of the microbial counts, except denitrifying bacteria, and all enzyme activities, except urease. Results indicated that soil compaction influences the community of aerobic microorganisms and their activity. This effect can alter nutrient cycling and reduce crop yields.

Glucose is arrhythmogenic in the anoxic-reoxygenated embryonic chick heart.

Unlike in adult heart, embryonic myocardium works at low PO2 and depends preferentially on glucose. Therefore, activity of the embryonic heart during anoxia and reoxygenation should be particularly affected by changes in glucose availability. Hearts excised from 4-d-old chick embryos were submitted in vitro to strictly controlled anoxia-reoxygenation transitions at glucose concentrations varying from 0 to 20 mmol/L. Spontaneous and regular heart contractions were detected optically as movements of the ventricle wall and instantaneous heart rate, amplitude of contraction, and velocities of contraction and relaxation were determined. Anoxia induced transient tachycardia and rapidly depressed contractile activity, whereas reoxygenation provoked a temporary and complete cardioplegia (oxygen paradox). In the presence of glucose, atrial rhythm became irregular during anoxia and chaotic-periodic during reoxygenation. The incidence of these arrhythmias depended on duration of anoxia, and no ventricular ectopic beats were observed. Removal of glucose or blockade of glycolysis suppressed arrhythmias. These results show similarities but also differences with respect to the adult heart. Indeed, glucose 1) delayed and anoxic contractile failure, shortened the reoxygenation-induced cardiac arrest, and improved the recovery of contractile activity; 2) attenuated stunning at 20 mmol/L but worsened it at 8 mmol/L; and 3) paradoxically, was arrhythmogenic during anoxia and reoxygenation, especially when present at the physiologic concentration of 8 mmol/L. The last named phenomenon seems to be characteristic of the young embryonic heart, and our findings underscore that fluctuations of glycolytic activity may play a role in the reactivity of the embryonic myocardium to anoxiareoxygenation transitions.

Effect of compaction on microbial activity and carbon and nitrogen transformations in two oxisols with different mineralogy

The use of machinery in agricultural and forest management activities frequently increases soil compaction, resulting in greater soil density and microporosity, which in turn reduces hydraulic conductivity and O2 and CO2 diffusion rates, among other negative effects. Thus, soil compaction has the potential to affect soil microbial activity and the processes involved in organic matter decomposition and nutrient cycling. This study was carried out under controlled conditions to evaluate the effect of soil compaction on microbial activity and carbon (C) and nitrogen (N) mineralization. Two Oxisols with different mineralogy were utilized: a clayey oxidic-gibbsitic Typic Acrustox and a clayey kaolinitic Xantic Haplustox (Latossolo Vermelho-Amarelo ácrico - LVA, and Latossolo Amarelo distrófico - LA, respectively, in the Brazil Soil Classification System). Eight treatments (compaction levels) were assessed for each soil type in a complete block design, with six repetitions. The experimental unit consisted of PVC rings (height 6 cm, internal diameter 4.55 cm, volume 97.6 cm³). The PVC rings were filled with enough soil mass to reach a final density of 1.05 and 1.10 kg dm-3, respectively, in the LVA and LA. Then the soil samples were wetted (0.20 kg kg-1 = 80 % of field capacity) and compacted by a hydraulic press at pressures of 0, 60, 120, 240, 360, 540, 720 and 900 kPa. After soil compression the new bulk density was calculated according to the new volume occupied by the soil. Subsequently each PVC ring was placed within a 1 L plastic pot which was then tightly closed. The soils were incubated under aerobic conditions for 35 days and the basal respiration rate (CO2-C production) was estimated in the last two weeks. After the incubation period, the following soil chemical and microbiological properties were detremined: soil microbial biomass C (C MIC), total soil organic C (TOC), total N, and mineral N (NH4+-N and NO3--N). After that, mineral N, organic N and the rate of net N mineralization was calculated. Soil compaction increased NH4+-N and net N mineralization in both, LVA and LA, and NO3--N in the LVA; diminished the rate of TOC loss in both soils and the concentration of NO3--N in the LA and CO2-C in the LVA. It also decreased the C MIC at higher compaction levels in the LA. Thus, soil compaction decreases the TOC turnover probably due to increased physical protection of soil organic matter and lower aerobic microbial activity. Therefore, it is possible to conclude that under controlled conditions, the oxidic-gibbsitic Oxisol (LVA) was more susceptible to the effects of high compaction than the kaolinitic (LA) as far as organic matter cycling is concerned; and compaction pressures above 540 kPa reduced the total and organic nitrogen in the kaolinitic soil (LA), which was attributed to gaseous N losses.

Oxygen tension controls the expression of the monocarboxylate transporter MCT4 in cultured mouse cortical astrocytes via a hypoxia-inducible factor-1α-mediated transcriptional regulation.

The monocarboxylate transporter MCT4 is a high capacity carrier important for lactate release from highly glycolytic cells. In the central nervous system, MCT4 is predominantly expressed by astrocytes. Surprisingly, MCT4 expression in cultured astrocytes is low, suggesting that a physiological characteristic, not met in culture conditions, is necessary. Here we demonstrate that reducing oxygen concentration from 21% to either 1 or 0% restored in a concentration-dependent manner the expression of MCT4 at the mRNA and protein levels in cultured astrocytes. This effect was specific for MCT4 since the expression of MCT1, the other astrocytic monocarboxylate transporter present in vitro, was not altered in such conditions. MCT4 expression was shown to be controlled by the transcription factor hypoxia-inducible factor-1α (HIF-1α) since under low oxygen levels, transfecting astrocyte cultures with a siRNA targeting HIF-1α largely prevented MCT4 induction. Moreover, the prolyl hydroxylase inhibitor dimethyloxalylglycine (DMOG) induced MCT4 expression in astrocytes cultured in presence of 21% oxygen. In parallel, glycolytic activity was enhanced by exposure to 1% oxygen as demonstrated by the increased lactate release, an effect dependent on MCT4 expression. Finally, MCT4 expression was found to be necessary for astrocyte survival when exposed for a prolonged period to 1% oxygen. These data suggest that a major determinant of astrocyte MCT4 expression in vivo is likely the oxygen tension. This could be relevant in areas of high neuronal activity and oxygen consumption, favouring astrocytic lactate supply to neurons. Moreover, it could also play an important role for neuronal recovery after an ischemic episode.

Incubation methods for assessing mineralizable nitrogen in soils under sugarcane

Considering nitrogen mineralization (N) of soil organic matter is a key aspect for the efficient management of N fertilizers in agricultural systems. Long-term aerobic incubation is the standard technique for calibrating the chemical extraction methods used to estimate the potentially mineralizable N in soil. However, the technique is laborious, expensive and time-consuming. In this context, the aims of this study were to determine the amount of soil mineralizable N in the 0-60 cm layer and to evaluate the use of short-term anaerobic incubation instead of long-term aerobic incubation for the estimation of net N mineralization rates in soils under sugarcane. Five soils from areas without previous N fertilization were used in the layers 0-20, 20-40 and 40-60 cm. Soil samples were aerobically incubated at 35 ºC for 32 weeks or anaerobically incubated (waterlogged) at 40 ºC for seven days to determine the net soil N mineralization. The sand, silt and clay contents were highly correlated with the indexes used for predicting mineralizable N. The 0-40 cm layer was the best sampling depth for the estimation of soil mineralizable N, while in the 40-60 cm layer net N mineralization was low in both incubation procedures. Anaerobic incubation provided reliable estimates of mineralizable N in the soil that correlated well with the indexes obtained using aerobic incubation. The inclusion of the pre-existing NH4+-N content improved the reliability of the estimate of mineralizable N obtained using anaerobic incubation.

Key electrophysiological, molecular, and metabolic signatures of sleep and wakefulness revealed in primary cortical cultures.

Although sleep is defined as a behavioral state, at the cortical level sleep has local and use-dependent features suggesting that it is a property of neuronal assemblies requiring sleep in function of the activation experienced during prior wakefulness. Here we show that mature cortical cultured neurons display a default state characterized by synchronized burst-pause firing activity reminiscent of sleep. This default sleep-like state can be changed to transient tonic firing reminiscent of wakefulness when cultures are stimulated with a mixture of waking neurotransmitters and spontaneously returns to sleep-like state. In addition to electrophysiological similarities, the transcriptome of stimulated cultures strikingly resembles the cortical transcriptome of sleep-deprived mice, and plastic changes as reflected by AMPA receptors phosphorylation are also similar. We used our in vitro model and sleep-deprived animals to map the metabolic pathways activated by waking. Only a few metabolic pathways were identified, including glycolysis, aminoacid, and lipids. Unexpectedly large increases in lysolipids were found both in vivo after sleep deprivation and in vitro after stimulation, strongly suggesting that sleep might play a major role in reestablishing the neuronal membrane homeostasis. With our in vitro model, the cellular and molecular consequences of sleep and wakefulness can now be investigated in a dish.

APPRAISAL OF THE SNAP MODEL FOR PREDICTING NITROGEN MINERALIZATION IN TROPICAL SOILS UNDER EUCALYPTUS

The Soil Nitrogen Availability Predictor (SNAP) model predicts daily and annual rates of net N mineralization (NNM) based on daily weather measurements, daily predictions of soil water and soil temperature, and on temperature and moisture modifiers obtained during aerobic incubation (basal rate). The model was based on in situ measurements of NNM in Australian soils under temperate climate. The purpose of this study was to assess this model for use in tropical soils under eucalyptus plantations in São Paulo State, Brazil. Based on field incubations for one month in three, NNM rates were measured at 11 sites (0-20 cm layer) for 21 months. The basal rate was determined in in situ incubations during moist and warm periods (January to March). Annual rates of 150-350 kg ha-1 yr-1 NNM predicted by the SNAP model were reasonably accurate (R2 = 0.84). In other periods, at lower moisture and temperature, NNM rates were overestimated. Therefore, if used carefully, the model can provide adequate predictions of annual NNM and may be useful in practical applications. For NNM predictions for shorter periods than a year or under suboptimal incubation conditions, the temperature and moisture modifiers need to be recalibrated for tropical conditions.

pH Sensitive surfactants from lysine: assessment of their cytotoxicity and environmental behavior

The toxicity and environmental behavior of new pH-sensitive surfactants from lysine are presented. Three different chemical structures are studied: surfactants with one amino acid and one alkyl chain, surfactants with two amino acids on the polar head and one alkyl chain, and gemini surfactants. The pH sensitivity of these compounds can be tuned by modifying their chemical structures. Cytotoxicity has been evaluated using erythrocytes and fibroblast cells. The toxic effects against these cells depend on the hydrophobicity of the molecules as well as their cationic charge density. The effect of hydrophobicity and cationic charge density on toxicity is different for each type of cells. For erythrocytes, the toxicity increases as hydrophobicity and charge density increases. Nevertheless, for fibroblasts cationic charge density affects cytotoxicity in the opposite way: the higher charge density, the lower the toxicity. The effect of the pH on hemolysis has been evaluated in detail. The aquatic toxicity was established using Daphnia magna. All surfactants yielded EC50 values considerably higher than that reported for cationic surfactants based on quaternary ammonium groups. Finally, their biodegradability was evaluated using the CO2 headspace test (ISO 14593). These lysine derivatives showed high levels of biodegradation under aerobic conditions and can be classified as"readily biodegradable compounds".

Microbiota present in cystic fibrosis lungs as revealed by whole genome sequencing.

Determination of the precise composition and variation of microbiota in cystic fibrosis lungs is crucial since chronic inflammation due to microorganisms leads to lung damage and ultimately, death. However, this constitutes a major technical challenge. Culturing of microorganisms does not provide a complete representation of a microbiota, even when using culturomics (high-throughput culture). So far, only PCR-based metagenomics have been investigated. However, these methods are biased towards certain microbial groups, and suffer from uncertain quantification of the different microbial domains. We have explored whole genome sequencing (WGS) using the Illumina high-throughput technology applied directly to DNA extracted from sputa obtained from two cystic fibrosis patients. To detect all microorganism groups, we used four procedures for DNA extraction, each with a different lysis protocol. We avoided biases due to whole DNA amplification thanks to the high efficiency of current Illumina technology. Phylogenomic classification of the reads by three different methods produced similar results. Our results suggest that WGS provides, in a single analysis, a better qualitative and quantitative assessment of microbiota compositions than cultures and PCRs. WGS identified a high quantity of Haemophilus spp. (patient 1) or Staphylococcus spp. plus Streptococcus spp. (patient 2) together with low amounts of anaerobic (Veillonella, Prevotella, Fusobacterium) and aerobic bacteria (Gemella, Moraxella, Granulicatella). WGS suggested that fungal members represented very low proportions of the microbiota, which were detected by cultures and PCRs because of their selectivity. The future increase of reads' sizes and decrease in cost should ensure the usefulness of WGS for the characterisation of microbiota.

Short-term overexpression of a constitutively active form of AMP-activated protein kinase in the liver leads to mild hypoglycemia and fatty liver.

AMP-activated protein kinase (AMPK) is a major therapeutic target for the treatment of diabetes. We investigated the effect of a short-term overexpression of AMPK specifically in the liver by adenovirus-mediated transfer of a gene encoding a constitutively active form of AMPKalpha2 (AMPKalpha2-CA). Hepatic AMPKalpha2-CA expression significantly decreased blood glucose levels and gluconeogenic gene expression. Hepatic expression of AMPKalpha2-CA in streptozotocin-induced and ob/ob diabetic mice abolished hyperglycemia and decreased gluconeogenic gene expression. In normal mouse liver, AMPKalpha2-CA considerably decreased the refeeding-induced transcriptional activation of genes encoding proteins involved in glycolysis and lipogenesis and their upstream regulators, SREBP-1 (sterol regulatory element-binding protein-1) and ChREBP (carbohydrate response element-binding protein). This resulted in decreases in hepatic glycogen synthesis and circulating lipid levels. Surprisingly, despite the inhibition of hepatic lipogenesis, expression of AMPKalpha2-CA led to fatty liver due to the accumulation of lipids released from adipose tissue. The relative scarcity of glucose due to AMPKalpha2-CA expression led to an increase in hepatic fatty acid oxidation and ketone bodies production as an alternative source of energy for peripheral tissues. Thus, short-term AMPK activation in the liver reduces blood glucose levels and results in a switch from glucose to fatty acid utilization to supply energy needs.

Assessment of physical activity with a pedometer and its relationship with VO2max among adolescents in Switzerland.

OBJECTIVES: In the absence of a gold standard, the assessment of physical activity in children remains difficult. To record physical activity with a pedometer and to examine to what extent it is correlated with VO2max. METHODS: Survey on physical activity and fitness; 233 Swiss adolescents aged 11 to 15 carried a pedometer (Pedoboy) during seven consecutive days. VO2max was estimated through an endurance shuttle run test. RESULTS: The physical activity recorded by the pedometer did not vary from one day to the other (p > 0.05). The physical activity was higher among boys than among girls (p < 0.001) and higher among younger adolescents (6th versus 8th grade; p < 0.001). The correlation between physical activity and estimated VO2max was 0.30 (p < 0.01). CONCLUSIONS: The use of a pedometer to assess physical activity over one entire week is feasible among adolescents. The record provided by the pedometer gives an objective measure of the usual physical activity and, as such, is relatively well correlated with aerobic capacity.

Effect of a lifestyle intervention on adiposity and fitness in socially disadvantaged subgroups of preschoolers: a cluster-randomized trial (Ballabeina).

OBJECTIVE: A multidimensional lifestyle intervention performed in 652 preschoolers (72% of migrant, 38% of low educational level (EL) parents) reduced body fat, but not BMI and improved fitness. The objective of this study is to examine whether the intervention was equally effective in children of migrant and/or low EL parents.¦METHODS: Cluster-randomized controlled single blinded trial, conducted in 2008/09 in 40 randomly selected preschools in Switzerland. The culturally tailored intervention consisted of a physical activity program and lessons on nutrition, media use and sleep. Primary outcomes included BMI and aerobic fitness. Secondary outcomes included %body fat, waist circumference and motor agility.¦RESULTS: Children of migrant parents benefitted similarly from the intervention compared to their counterparts (p for interaction≥ 0.09). However, children of low EL parents benefitted less, although these differences did not reach statistical significance (p for interaction≥ 0.06). Average intervention effect sizes for BMI were -0.10, -0.05, -0.11 and 0.04 kg/m(2) and for aerobic fitness were 0.55, 0.20, 0.37 and -0.05 stages for children of non-migrant, migrant, middle/high EL and low EL parents, respectively.¦CONCLUSIONS: This intervention was similarly effective among preschoolers of migrant parents compared to their counterparts, while children of low EL parents benefitted less.