Differences in SOM Decomposition and Temperature Sensitivity among Soil Aggregate Size Classes in a Temperate Grasslands

The principle of enzyme kinetics suggests that the temperature sensitivity (Q₁₀) of soil organic matter (SOM) decomposition is inversely related to organic carbon (C) quality, i.e., the C quality-temperature (CQT) hypothesis. We tested this hypothesis by performing laboratory incubation experiments with bulk soil, macroaggregates (MA, 250–2000 μm), microaggregates (MI, 53–250 μm), and mineral fractions (MF, <53 μm) collected from an Inner Mongolian temperate grassland. The results showed that temperature and aggregate size significantly affected on SOM decomposition, with notable interactive effects (P<0.0001). For 2 weeks, the decomposition rates of bulk soil and soil aggregates increased with increasing incubation temperature in the following order: MA>MF>bulk soil >MI(P <0.05). The Q₁₀ values were highest for MA, followed (in decreasing order) by bulk soil, MF, and MI. Similarly, the activation energies (E_a) for MA, bulk soil, MF, and MI were 48.47, 33.26, 27.01, and 23.18 KJ mol⁻¹, respectively. The observed significant negative correlations between Q₁₀ and C quality index in bulk soil and soil aggregates (P<0.05) suggested that the CQT hypothesis is applicable to soil aggregates. Cumulative C emission differed significantly among aggregate size classes (P <0.0001), with the largest values occurring in MA (1101 μg g⁻¹), followed by MF (976 μg g⁻¹) and MI (879 μg g⁻¹). These findings suggest that feedback from SOM decomposition in response to changing temperature is closely associated withsoil aggregation and highlights the complex responses of ecosystem C budgets to future warming scenarios.     

Modulation of the Voltage-gated Potassium Channel (Kv4.3) and the Auxiliary Protein (KChIP3) Interactions by the Current Activator NS5806

KChIP3 (potassium channel interacting protein 3) is a calcium-binding protein that binds at the N terminus of the Kv4 voltage-gated potassium channel through interactions at two contact sites and has been shown to regulate potassium current gating kinetics as well as channel trafficking in cardiac and neuronal cells. Using fluorescence spectroscopy, isothermal calorimetry, and docking simulations we show that the novel potassium current activator, NS5806, binds at a hydrophobic site on the C terminus of KChIP3 in a calcium-dependent manner, with an equilibrium dissociation constant of 2–5 μm in the calcium-bound form. We further determined that the association between KChIP3 and the hydrophobic N terminus of Kv4.3 is calcium-dependent, with an equilibrium dissociation constant in the apo-state of 70 ± 3 μm and 2.7 ± 0.1 μm in the calcium-bound form. NS5806 increases the affinity between KChIP3 and the N terminus of Kv4.3 (K_d = 1.9 ± 0.1 μm) in the presence and absence of calcium. Mutation of Tyr-174 or Phe-218 on KChIP3 abolished the enhancement of Kv4.3 site 1 binding in the apo-state, highlighting the role of these residues in drug and K4.3 binding. Kinetic studies show that NS5806 decreases the rate of dissociation between KChIP3 and the N terminus of K_V4.3. Overall, these studies support the idea that NS5806 directly interacts with KChIP3 and modulates the interactions between this calcium-binding protein and the T1 domain of the Kv4.3 channels through reorientation of helix 10 on KChIP3.     

Environmental manipulation treatment effects on the reactivity of water-soluble phenolics in a subalpine tundra ecosystem

Low soil organic matter content and limited soil water holding are the major natural constraint of dryland cropping on sandy soils in the Quebec boreal regions. We conducted a 3-yr (1994–1996) study in a boreal sandy soil, Ferro-Humic Podzol (Spodosols), to determine the potential of Sphagnum peat for improving soil organic matter (SOM), water holding capacity, bulk density (BD), plant leaf nutrient status, and potato and barley yields. The cropping was a rotation of 2-yr potato (Solanum tuberosum L. Superior) and 1-yr barley (Hordeum vulgare L. Chapais). The treatments consisted of Sphagnum peat at rates of 0, 29, 48, and 68 Mg ha^–1 3-yr^–1 on a dry weight basis, and granular N-P-K fertilizers (12-7.5-7) at rates of 1.4, 1.6, and 1.8 Mg ha^–1 yr^–1, respectively, arranged in a split-block design. The peat-amended soils were higher in water content (SWC), SOM and total porosity but lower in BD and N than neighboring non-peat soils (P < 0.05). Effects of peat and fertilizer treatments and their interaction were significant on potato leaf N, Ca, Mg, and P, tuber yield, dry weight, harvested N and tuber specific gravity (P < 0.05), depending on year. Potato tuber yield and N increased simultaneously up to 30% (compared to the control), and were significantly correlated with SWC, SOM, BD, and NO₃-N (–0.52 r 0.80). In the 3rd year, the linear effect of peat treatments was significant on barley grain yield. In 1995 there was a decline of 4.5–7.3% of SOM of the previous year level. It is suggested that Sphagnum peat at a rate of 48 Mg ha^–1 had the potential for improving sandy soil productivity. A longer-term investigation of soil water, N, SOM pool and crop yield changes is necessary to better understand the physical, chemical and biological processes of peat in cropping systems and to maximize the benefits of peat applications.     

Dynamics of Soil Organic Carbon and Aggregate Stability with Grazing Exclusion in the Inner Mongolian Grasslands

Grazing exclusion (GE) has been deemed as an important approach to enhance the soil carbon storage of semiarid grasslands in China; however, it remains unclear how different organic carbon (OC) components in soils vary with the duration of GE. Here, we observed the changing trends of different OC components in soils with increased GE duration in five grassland succession series plots, ranging from free grazing to 31-year GE. Specifically, we measured microbial biomass carbon (MBC), easily oxidizable OC (EOC), water-soluble OC (WSOC), and OC in water stable aggregates (macroaggregates [250–2000 μm], microaggregates [53–250 μm], and mineral fraction [< 53 μm]) at 0–20 cm soil depths. The results showed that GE significantly enhanced EOC and WSOC contents in soils, but caused a decline of MBC at the three decade scale. Macroaggregate content (F = 425.8, P < 0.001), OC stored in macroaggregates (F = 84.1, P < 0.001), and the mean weight diameter (MWD) of soil aggregates (F = 371.3, P < 0.001) increased linearly with increasing GE duration. These findings indicate that OC stored in soil increases under three-decade GE with soil organic matter (SOM) stability improving to some extent. Long-term GE practices enhance the formation of soil aggregates through higher SOM input and an exclusion of animal trampling. Therefore, the practice of GE may be further encouraged to realize the soil carbon sequestration potential of semi-arid grasslands, China.     

Fertilization regimes affect the soil biological characteristics of a sudangrass and ryegrass rotation system

The sudangrass (Sorghum sudanense) and ryegrass (Lolium multiflorum L.) rotation is an intensive and new cropping system in Central China. Nutrient management practices in this rotation system may influence soil fertility, the important aspects of which are soil biological properties and quality. As sensitive soil biological properties and quality indicators, soil microbial community activity, microbial biomass, enzyme activities, soil organic matter (SOM) and total N resulting from different fertilization regimes in this rotation system were studied through a four-year field experiment from April 2005 to May 2009. Treatments included control (CK), fertilizer phosphorus and potassium (PK), fertilizer nitrogen and potassium (NK), fertilizer nitrogen and phosphorus (NP) and a fertilizer nitrogen, phosphorus and potassium combination (NPK). Soil microbial community activities in the NK, NP and NPK treatments were significantly lower than those in the CK and PK treatments after the sudangrass and ryegrass trial. The highest microbial biomass C, microbial biomass N, SOM, total N, sucrase and urease activities were found in the NPK treatment, and these soil quality indicators were significantly higher in the NK, NP and NPK treatments than in the PK and CK treatments. Soil microbial biomass and enzyme activities were positively associated with SOM in the sudangrass and ryegrass rotation system, indicating that fertilization regimes, especially N application, reduced microbial community activity in the soil. Proper fertilization regimes will increase microbial biomass, enzyme activity and SOM and improve soil fertility.     

Echinacoside protects adenine-induced uremic rats from sciatic nerve damage by up-regulating α-Klotho

Objectives:To investigate the therapeutic effect of Echinacoside on uremia-induced sciatic nerve injury and explore the specific molecular mechanism and role of α-Klotho.Methods:SD rats were given continuous gavage of adenine to prepare a uremia-induced sciatic nerve injury model. The model was given either Echinacoside or α-Klotho by gavage. Histopathological changes of kidney and sciatic nerve were detected by H&E staining. The changes of creatinine, urea nitrogen, and urine protein were detected by biochemical detection. The changes of IL-1β and IL-18 were detected by ELISA. Nerve activity-related indicators were detected by biochemical detection. Changes in related mRNA and protein expression were detected by qPCR and western blot.Results:Creatinine, urea nitrogen, urine protein, and malondialdehyde (MDA) in the model group were significantly increased and inhibited by Echinacoside and α-Klotho treatment with Echinacoside dose-dependence. Meanwhile, the activities of ATP concentration, potassium adenosine triphosphate (Na+, K+ ATPase), succinate dehydrogenase (SDH), glutathione peroxidase (GSH-Px), and superoxide dismutase (SOD) showed opposite trends.Conclusions:Echinacoside can significantly relieve uremia-induced sciatic nerve injury in rats. Its specific molecular mechanism is related to the inhibition of the classical cellular pyroptosis pathway, which is likely achieved by promoting α-Klotho expression.     

Stabilization mechanisms of soil organic matter: Implications for C-saturation of soils

The relationship between soil structure and the ability of soil to stabilize soil organic matter (SOM) is a key element in soil C dynamics that has either been overlooked or treated in a cursory fashion when developing SOM models. The purpose of this paper is to review current knowledge of SOM dynamics within the framework of a newly proposed soil C saturation concept. Initially, we distinguish SOM that is protected against decomposition by various mechanisms from that which is not protected from decomposition. Methods of quantification and characteristics of three SOM pools defined as protected are discussed. Soil organic matter can be: (1) physically stabilized, or protected from decomposition, through microaggregation, or (2) intimate association with silt and clay particles, and (3) can be biochemically stabilized through the formation of recalcitrant SOM compounds. In addition to behavior of each SOM pool, we discuss implications of changes in land management on processes by which SOM compounds undergo protection and release. The characteristics and responses to changes in land use or land management are described for the light fraction (LF) and particulate organic matter (POM). We defined the LF and POM not occluded within microaggregates (53–250 m sized aggregates as unprotected. Our conclusions are illustrated in a new conceptual SOM model that differs from most SOM models in that the model state variables are measurable SOM pools. We suggest that physicochemical characteristics inherent to soils define the maximum protective capacity of these pools, which limits increases in SOM (i.e. C sequestration) with increased organic residue inputs.     

Rhizosphere respiration varies with plant species and phenology: A greenhouse pot experiment

Assessment of particulate (>53-m) and mineral-associated (<53-m) soil organic matter (SOM) fractions is a useful approach to understand the dynamic of organic matter in soils. This study aimed to compare the long-term (9-yr) effects of no-tillage (NT) and conventional tillage (CT) on C and N stocks in the two above mentioned organic fractions in a Brazilian Acrisol. The degree of SOM humification, which has been associated with the concentration of semiquinone-type free radicals (`spin') determined by electron spin resonance (ESR), was also evaluated. Soil under no-tillage had 7.55 Mg ha<sup>–1</sup> (25%) more C and 741 kg ha<sup>–1</sup> (29%) more N than conventionally tilled soil in the 0–175-mm depth. Both particulate and mineral-associated SOM increased in the no-tilled soil. The increase of C and N stocks in the mineral-associated SOM accounted for 75% and 91% of the difference in total soil C and N stocks between NT and CT, respectively. Averaged across tillage systems, C and N stocks were respectively 4.6 and 16.8 times higher in the mineral-associated SOM than in particulate SOM. The higher C and N stocks were associated with greater recalcitrance of mineral-associated SOM to biological decomposition, resulting, probably, from its interaction with variable charge minerals. This is corroborated by a positive relationship between concentrations of C and iron oxides and kaolinite in the 53–20, 20–2 and <2-m particle size classes, of the 0–25-mm soil layer. The degree of SOM humification, assessed by ESR, decreased in both the 53–20 and 20–2-m fractions under NT. However, it was unaffected by tillage in the <2-m fraction, which normally presented the lowest `spin' concentration. Since quality as well as quantity of SOM improved in the no-tillage soil, adoption of this system is highly recommended for amelioration of degraded tropical and subtropical soils.     

Effects of Different Organic Manures on the Biochemical and Microbial Characteristics of Albic Paddy Soil in a Short-Term Experiment

This study aimed to evaluate the effects of chemical fertilizer (NPK), NPK with livestock manure (NPK+M), NPK with straw (NPK+S), and NPK with green manure (NPK+G) on soil enzyme activities and microbial characteristics of albic paddy soil, which is a typical soil with low productivity in China. The responses of extracellular enzyme activities and the microbial community diversity (determined by phospholipid fatty acid analysis [PLFA] and denaturing gradient gel electrophoresis [DGGE]) were measured. The results showed that NPK+M and NPK+S significantly increased rice yield, with NPK+M being approximately 24% greater than NPK. The NPK+M significantly increased soil organic carbon (SOC) and available phosphate (P) and enhanced phosphatase, β-cellobiosidase, L-leucine aminopeptidase and urease activities. The NPK+S significantly increased SOC and available potassium (K) and significantly enhanced N-acetyl-glucosamidase, β-xylosidase, urease, and phenol oxidase activities. The NPK+G significantly improved total nitrogen (N), ammonium N, available P, and N-acetyl-glucosamidase activity. The PLFA biomass was highest under NPK+S, followed by NPK+M and NPK+G treatments. Principal component analysis (PCA) of the PLFA indicated that soils with NPK+M and NPK+S contained higher proportions of unsaturated and cyclopropane fatty acids (biomarkers of fungi and gram-negative bacteria) and soil under NPK+G contained more straight chain saturated fatty acids (representing gram-positive bacteria). PCA of the DGGE patterns showed that organic amendments had a greater influence on fungal community. Cluster analysis of fungal DGGE patterns revealed that NPK+G was clearly separated. Meanwhile, the bacterial community of NPK+M treatment was the most distinct. RDA analysis revealed changes of microbial community composition mostly depended on β-xylosidase, β-cellobiosidase activities, total N and available K contents. The abundances of gram-negative bacterial and fungal PLFAs probably effective in improving fertility of low-yield albic paddy soil because of their significant influence on DGGE profile.     

Changes in Forest Soil Properties in Different Successional Stages in Lower Tropical China

Background

Natural forest succession often affects soil physical and chemical properties. Selected physical and chemical soil properties were studied in an old-growth forest across a forest successional series in Dinghushan Nature Reserve, Southern China.

Methodology/Principal Findings

The aim was to assess the effects of forest succession change on soil properties. Soil samples (0–20 cm depth) were collected from three forest types at different succession stages, namely pine (Pinus massoniana) forest (PMF), mixed pine and broadleaf forest (PBMF) and monsoon evergreen broadleaf forest (MEBF), representing early, middle and advanced successional stages respectively. The soil samples were analyzed for soil water storage (SWS), soil organic matter (SOM), soil microbial biomass carbon (SMBC), pH, NH₄ ⁺-N, available potassium (K), available phosphorus (P) and microelements (available copper (Cu), available zinc (Zn), available iron (Fe) and available boron (B)) between 1999 and 2009. The results showed that SWS, SOM, SMBC, Cu, Zn, Fe and B concentrations were higher in the advanced successional stage (MEBF stage). Conversely, P and pH were lower in the MEBF but higher in the PMF (early successional stage). pH, NH₄ ⁺-N, P and K declined while SOM, Zn, Cu, Fe and B increased with increasing forest age. Soil pH was lower than 4.5 in the three forest types, indicating that the surface soil was acidic, a stable trend in Dinghushan.

Conclusion/Significance

These findings demonstrated significant impacts of natural succession in an old-growth forest on the surface soil nutrient properties and organic matter. Changes in soil properties along the forest succession gradient may be a useful index for evaluating the successional stages of the subtropical forests. We caution that our inferences are drawn from a pseudo-replicated chronosequence, as true replicates were difficult to find. Further studies are needed to draw rigorous conclusions regarding on nutrient dynamics in different successional stages of forest.     

Partitioning of CH4 and CO2 Production Originating from Rice Straw,Soil and Root Organic Carbon in Rice Microcosms

Flooded rice fields are an important source of the greenhouse gas CH₄. Possible carbon sources for CH₄ and CO₂ production in rice fields are soil organic matter (SOM), root organic carbon (ROC) and rice straw (RS), but partitioning of the flux between the different carbon sources is difficult. We conducted greenhouse experiments using soil microcosms planted with rice. The soil was amended with and without ¹³C-labeled RS, using two ¹³C-labeled RS treatments with equal RS (5 g kg⁻¹ soil) but different δ¹³C of RS. This procedure allowed to determine the carbon flux from each of the three sources (SOM, ROC, RS) by determining the δ¹³C of CH₄ and CO₂ in the different incubations and from the δ¹³C of RS. Partitioning of carbon flux indicated that the contribution of ROC to CH₄ production was 41% at tillering stage, increased with rice growth and was about 60% from the booting stage onwards. The contribution of ROC to CO₂ was 43% at tillering stage, increased to around 70% at booting stage and stayed relatively constant afterwards. The contribution of RS was determined to be in a range of 12–24% for CH₄ production and 11–31% for CO₂ production; while the contribution of SOM was calculated to be 23–35% for CH₄ production and 13–26% for CO₂ production. The results indicate that ROC was the major source of CH₄ though RS application greatly enhanced production and emission of CH₄ in rice field soil. Our results also suggest that data of CH₄ dissolved in rice field could be used as a proxy for the produced CH₄ after tillering stage.     

Manure and mineral fertilization change enzyme activity and bacterial community in millet rhizosphere soils

Fertilization is a key agricultural practice for increasing millet yields and influencing soil properties, enzyme activities and rhizosphere bacterial communities. High throughput Illumina sequencing of the 16S rDNA was applied to compare the bacterial community structures and diversities among six different soil samples. The experiments involved the following: no fertilizer (CK), phosphate (P) and potassium (K) plus organic manure (M) (PKM), nitrogen (N) and K plus M (NKM), NPM, NPK and NPKM fertilization. The results showed that the NPKM fertilization of the millet field had a maximal yield of 3617 kg ha^?1 among the six different treatments. The abundances of the Actinobacteria and Bacteroidetes phyla, especially the Devosia, Mycobacterium, Opitutus and Chitinophaga genera, were higher in NPKM than those in the other samples. Redundancy analysis showed that the soil organic matter (SOM), available phosphorus (AP), and urease (UR) activity were significantly correlated with bacterial communities, while SOM and AP were strongly correlated with soil enzyme activities. Pearson’s correlation showed that the available nitrogen was strongly correlated with Devosia and Mycobacterium, and SOM was strongly correlated with Opitutus and Chitinophaga. Besides, catalase was significantly related to Iamia, the UR was significantly related to Devosia, phosphatase was significantly related to Luteimonas and Chitinophaga. Based on the soil quality and millet yield, NPKM treatment was a better choice for the millet field fertilization practices. These findings provide a better understanding of the importance of fertilization in influencing millet yield, soil fertility and microbial diversity, and they lead to a choice of scientific fertilization practices for sustainable development of the agroecosystem.     

Concentrations and resorption patterns of 13 nutrients in different plant functional types in the karst region of south-western China

Background and Aims

Elucidating the stoichiometry and resorption patterns of multiple nutrients is an essential requirement for a holistic understanding of plant nutrition and biogeochemical cycling. However, most studies have focused on nitrogen (N) and phosphorus (P), and largely ignored other nutrients. The current study aimed to determine relationships between resorption patterns and leaf nutrient status for 13 nutrient elements in a karst vegetation region.

Methods

Plant and soil samples were collected from four vegetation types in the karst region of south-western China and divided into eight plant functional types. Samples of newly expanded and recently senesced leaves were analysed to determine concentrations of boron (B), calcium (Ca), copper (Cu), iron (Fe), potassium (K), magnesium (Mg), manganese (Mn), molybdenum (Mo), N, sodium (Na), P, sulphur (S) and zinc (Zn).

Key Results

Nutrient concentrations of the karst plants were lower than those normally found in other regions of China and the rest of the world, and plant growth was mainly limited by P. Overall, four nutrients revealed resorption [N (resorption efficiency 34·6 %), P (48·4 %), K (63·2 %) and Mg (13·2 %)], seven nutrients [B (–16·1 %), Ca (–44·0 %), Cu (–14·5 %), Fe (–205·5 %), Mn (–72·5 %), Mo (–35·6 %) and Zn (–184·3 %)] showed accumulation in senesced leaves and two nutrients (Na and S) showed no resorption or accumulation. Resorption efficiencies of K and Mg and accumulation of B, Ca, Fe and Mn differed among plant functional types, and this strongly affected litter quality. Resorption efficiencies of N, P and K and accumulation of Ca and Zn increased with decreasing concentrations of these nutrients in green leaves. The N:P, N:K and N:Mg ratios in green leaves predicted resorption proficiency for N, K and Mg, respectively.

Conclusions

The results emphasize the fact that nutrient resorption patterns strongly depend on element and plant functional type, which provides new insights into plant nutrient use strategies and nutrient cycling in karst ecosystems.     

Components of Sodium and Chloride Flux Across Toad Bladder

The effect of transepithelial potential difference (ψ) on Na and Cl flux across toad bladder was assessed by measuring isotopic flux between identical media at various values of ψ. The contribution of edge damage to ionic permeability was eliminated, resulting in relatively high spontaneous ψ (-97 ±4 mv) and low electrical conductance g. Bidirectional Na fluxes were measured simultaneously. Unidirectional Cl fluxes were measured in paired hemibladders at ψ = 0 mv or -97 mv. Net Na flux J_Na, at ψ = 0 mv, was slightly less than short-circuit current (SCC). At ψ = -97 mv, J_Na averaged 17% of SCC, and was sometimes zero. ΔJ_Na/Δψ (= g₊) averaged 60% of g between -97 mv and +75 mv; at -150 mv, g₊ fell, indicating rectification. Analysis of unidirectional Na fluxes indicates low passive conductance (1.5 μmho/mg wet weight), a bidirectional, electrically neutral flux of approximately 0.13 μa/mg, and relatively large conductance of the active transport path at ψ ≥ -97 mv. The absence of appreciable transstimulation of serosal (S)-to-mucosal (M) Na flux (in response to increasing mucosal Na concentration) indicates that the electrically neutral flux is not exchange diffusion in the usual sense. Analysis of Cl fluxes indicates similar values for passive conductance and neutral flux, suggesting linked neutral flux of Na and Cl. Either the electromotive force of the Na pump E, its conductance g_a, or both are strong functions of ψ. The product of these two quantities, Eg_a, is a measure of the “transport capacity” at any given value of ψ, independent of the direct effect of ψ on J_Na through the pump path. Eg_a varies with ψ. Hence estimation of the net Na flux or current at any one value of ψ, including ψ = 0, fails to reveal the maximal transport capacity of the pump, its resting electromotive force (when J_Na = 0 through the pump), or the dependence of transport capacity on potential.     

Path Analysis to Identify Factors Influencing Health Skills and Behaviors in Adolescents: A Cross-Sectional Survey

Background

Studies conducted in the past mostly rely on models of functional health literacy in adult populations. However, such models do not satisfy the need for health intervention in adolescents. The identification of key factors influencing adolescents'' health literacy is essential in developing effective prevention and intervention measures. This study aimed to test a theoretical model of predictors on health skills and health behaviors in adolescents.

Methods

A cross-sectional survey was conducted in Guangdong using a multi-stage stratified cluster sample design. A representative random sample of 3821 students aged 13–25 years was selected using multi-stage stratified cluster sampling. The path analysis was used to test a hypothesized model of health literacy.

Results

The path analysis showed that knowledge of infectious disease (β = 0.26), health skills (β = 0.22), health concept (β = 0.20), general health knowledge (β = 0.15), gender (β = 0.12), and school performance (β = 0.06) had positive direct effect on health behaviors in adolescents. The explanatory variables accounted for 43% of the variance in explaining health behaviors. Knowledge of infectious disease (β = 0.30), health concept (β = 0.17), general health knowledge (β = 0.13), and school performance (β = 0.05) had positive indirect effect on health behaviors through the impacts on health skills.

Conclusion

This study identified several direct and indirect factors influencing health skills and health behaviors in adolescents. These findings will assist health professionals designing effective health interventions that aim to improve health skills and health behaviors in adolescents.     

Do the Levels of Maternal Plasma Trace Elements Affect Fetal Nuchal Translucency Thickness?

Objective

Fetal nuchal translucency (NT) thickness is an important marker for prenatal screening; however, studies focusing on the correlation between maternal trace element levels and NT thickness are limited. The aim of this study was to evaluate maternal trace element levels during the first trimester and to investigate the association between maternal trace element levels and fetal NT thickness.

Methods

In total, 113 samples were obtained from singleton pregnant women. Maternal plasma samples were collected in the first trimester of gestation. Plasma trace element levels were measured using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Nuchal translucency thickness was measured using ultrasonography at 10–14 weeks of gestation.

Results

We found that maternal plasma potassium (K) levels had a significant negative correlation with both NT (r = -0.230, p < 0.05) and NT Multiples of the Median (NT MoM) (r = -0.206, p < 0.05). After adjustment for potential confounders, log-transformed maternal plasma potassium levels in the first trimester were significantly associated with fetal NT (NT MoM: β = -0.68, p < 0.05; NT: β = -1.20, p < 0.01). Although not statistically significant, the As, Hg and Pb levels in maternal plasma were positively correlated with NT, and the Mg, Cu, Zn, Na and Ca levels were negatively correlated with NT.

Conclusion

Maternal plasma K levels during the first trimester appeared to be associated with NT thickness. The essential elements tended to decrease NT thickness, and non-essential elements tended to increase it.     

GEOTROPIC ORIENTATION IN ARTHROPODS : I. MALACOSOMA LARV?.

The geotropic orientation of caterpillars of Malacosoma americana during progression upon a surface inclined at angle α to the horizontal is such that the path makes an average angle θ upward on the plane, of a magnitude proportional to log sin α. More precisely, the product (sin α) (sin θ) is constant. This is traced to the fluctuation of the pull of the head region upon the lateral musculature of the upper side during the side to side swinging implicated in progression.     

α1-AMP-Activated Protein Kinase Regulates Hypoxia-Induced Na,K-ATPase Endocytosis via Direct Phosphorylation of Protein Kinase Cζ

Hypoxia promotes Na,K-ATPase endocytosis via protein kinase Cζ (PKCζ)-mediated phosphorylation of the Na,K-ATPase α subunit. Here, we report that hypoxia leads to the phosphorylation of 5′-AMP-activated protein kinase (AMPK) at Thr172 in rat alveolar epithelial cells. The overexpression of a dominant-negative AMPK α subunit (AMPK-DN) construct prevented the hypoxia-induced endocytosis of Na,K-ATPase. The overexpression of the reactive oxygen species (ROS) scavenger catalase prevented hypoxia-induced AMPK activation. Moreover, hypoxia failed to activate AMPK in mitochondrion-deficient ρ⁰-A549 cells, suggesting that mitochondrial ROS play an essential role in hypoxia-induced AMPK activation. Hypoxia-induced PKCζ translocation to the plasma membrane and phosphorylation at Thr410 were prevented by the pharmacological inhibition of AMPK or by the overexpression of the AMPK-DN construct. We found that AMPK α phosphorylates PKCζ on residue Thr410 within the PKCζ activation loop. Importantly, the activation of AMPK α was necessary for hypoxia-induced AMPK-PKCζ binding in alveolar epithelial cells. The overexpression of T410A mutant PKCζ prevented hypoxia-induced Na,K-ATPase endocytosis, confirming that PKCζ Thr410 phosphorylation is essential for this process. PKCζ activation by AMPK is isoform specific, as small interfering RNA targeting the α1 but not the α2 catalytic subunit prevented PKCζ activation. Accordingly, we provide the first evidence that hypoxia-generated mitochondrial ROS lead to the activation of the AMPK α1 isoform, which binds and directly phosphorylates PKCζ at Thr410, thereby promoting Na,K-ATPase endocytosis.When exposed to low oxygen levels (hypoxia), cells develop adaptative strategies to maintain adequate levels of ATP (21). These strategies include increasing the efficiency of energy-producing pathways, mostly through anaerobic glycolysis, while decreasing energy-consuming processes such as Na,K-ATPase activity (30). Alveolar hypoxia occurs in many respiratory disorders, and it has been shown to decrease epithelial active Na⁺ transport, leading to impaired fluid reabsorption (37, 41, 42). Active Na⁺ transport and, thus, alveolar fluid reabsortion are effected mostly via apical sodium channels and the basolateral Na,K-ATPase (32, 38, 42). We have reported previously that hypoxia inhibits Na,K-ATPase activity by promoting its endocytosis from the plasma membrane by a mechanism that requires the generation of mitochondrial reactive oxygen species (ROS) and the phosphorylation of the Na,K-ATPase α subunit at Ser18 by protein kinase Cζ (PKCζ) (8, 9).The 5′-AMP-activated protein kinase (AMPK) is a heterotrimeric Ser/Thr kinase composed of a catalytic α subunit and regulatory β and γ subunits. Both isoforms of the AMPK catalytic subunit (α1 and α2) form complexes with noncatalytic subunits. The α1 subunit is ubiquitously expressed, whereas the α2 subunit isoform is expressed predominantly in tissues like the liver, heart, and skeletal muscle (36). The α1 and α2 subunit isoforms have ∼90% homology in their N-terminal catalytic domains and ∼60% homology in their C-terminal domains (36), suggesting that they may have distinct downstream targets (31). AMPK activation requires phosphorylation at Thr172 in the activation loop of the α subunit by upstream kinases (12, 19). Findings from recent studies suggest that AMPK is an important signaling intermediary in coupling ion transport and metabolism (15). Indeed, it has been reported that the pharmacological activation of AMPK inhibits amiloride- and ouabain-sensitive epithelial Na⁺ transport (15). Moreover, the activities of the epithelial Na⁺ channel (ENaC) (2, 17), the Na,K-ATPase (40), and the cystic fibrosis transmembrane conductance regulator (17) have been shown to be inhibited by AMPK. Here, we provide evidence that hypoxia, via mitochondrial ROS, leads to AMPK activation and that AMPK binds to and directly phosphorylates PKCζ in an isoform-specific manner, thus promoting Na,K-ATPase endocytosis in alveolar epithelial cells (AEC).     

Altered PI3-Kinase/Akt Signalling in Skeletal Muscle of Young Men with Low Birth Weight

Background

Low birth weight (LBW) is associated with increased future risk of insulin resistance and type 2 diabetes mellitus. The underlying molecular mechanisms remain poorly understood. We have previously shown that young LBW men have reduced skeletal muscle expression of PI3K p85α regulatory subunit and p110β catalytic subunit, PKCζ and GLUT4 in the fasting state. The aim of this study was to determine whether insulin activation of the PI3K/Akt and MAPK signalling pathways is altered in skeletal muscle of young adult men with LBW.

Methods

Vastus lateralis muscle biopsies were obtained from 20 healthy 19-yr old men with BW</ = 10^th percentile for gestational age (LBW) and 20 normal birth weight controls (NBW), matched for physical fitness and whole-body glucose disposal, prior to (fasting state) and following a 4-hr hyperinsulinemic euglycemic clamp (insulin stimulated state). Expression and phosphorylation of selected proteins was determined by Western blotting.

Principal Findings

Insulin stimulated expression of aPKCζ (p<0.001) and Akt1 (p<0.001) was decreased in muscle of LBW men when compared to insulin stimulated controls. LBW was associated with increased insulin stimulated levels of IRS1 (p<0.05), PI3K p85α (p<0.001) and p110β (p<0.05) subunits, while there was no significant change in these proteins in insulin stimulated control muscle. In addition LBW had reduced insulin stimulated phospho-Akt (Ser 473) (p<0.01), indicative of reduced Akt signalling. Insulin stimulated expression/phosphorylation of all the MAPK proteins studied [p38 MAPK, phospho-p38 MAPK (Thr180/Tyr182), phospho-ERK (Thr 202/Tyr204), JNK1, JNK2 and phospho-JNK (Thr 183/Tyr185)] was not different between groups.

Conclusions

We conclude that altered insulin activation of the PI3K/Akt but not the MAPK pathway precedes and may contribute to development of whole-body insulin resistance and type 2 diabetes in men with LBW.     

Low Na,High K Diet and the Role of Aldosterone in BK-Mediated K Excretion

A low Na, high K diet (LNaHK) is associated with a low rate of cardiovascular (CV) disease in many societies. Part of the benefit of LNaHK relies on its diuretic effects; however, the role of aldosterone (aldo) in the diuresis is not understood. LNaHK mice exhibit an increase in renal K secretion that is dependent on the large, Ca-activated K channel, (BK-α with accessory BK-β4; BK-α/β4). We hypothesized that aldo causes an osmotic diuresis by increasing BK-α/β4-mediated K secretion in LNaHK mice. We found that the plasma aldo concentration (P[aldo]) was elevated by 10-fold in LNaHK mice compared with control diet (Con) mice. We subjected LNaHK mice to either sham surgery (sham), adrenalectomy (ADX) with low aldo replacement (ADX-LA), or ADX with high aldo replacement (ADX-HA). Compared to sham, the urinary flow, K excretion rate, transtubular K gradient (TTKG), and BK-α and BK-β4 expressions, were decreased in ADX-LA, but not different in ADX-HA. BK-β4 knockout (β4KO) and WT mice exhibited similar K clearance and TTKG in the ADX-LA groups; however, in sham and ADX-HA, the K clearance and TTKG of β4KO were less than WT. In response to amiloride treatment, the osmolar clearance was increased in WT Con, decreased in WT LNaHK, and unchanged in β4KO LNaHK. These data show that the high P[aldo] of LNaHK mice is necessary to generate a high rate of BK-α/β4-mediated K secretion, which creates an osmotic diuresis that may contribute to a reduction in CV disease.