Real-time monitoring of adhesion and aggregation of platelets using thickness shear mode (TSM) sensor

Hemostasis is required to maintain vascular system integrity, but thrombosis, formation of a clot in a blood vessel, is one of the largest causes of morbidity and mortality in the industrialized world. Novel clinical and research tools for characterizing the hemostatic system are of continued interest, and the object of this research is to test the hypothesis that clinically relevant platelet function can be monitored using an electromechanical sensor. A piezoelectric thickness shear mode (TSM) biosensor coated with collagen-I fibers to promote platelet activation and adhesion was developed and tested for sensitivity to detect these primary events. Magnitude and frequency response of the sensor were monitored under static conditions at 37 degrees C, using platelet-rich plasma (PRP), and PRP with adenosine diphosphate (ADP), a clinical aggregation inhibitor (abciximab), or a collagen binding inhibitor. Sensors loaded with PRP exhibited a 3-stage response; no significant change in response for the first 20 min (Stage-1), followed by a larger drop in response (Stage-2) and subsequently, response gradually increased (Stage-3). Exogenous ADP stimulated an immediate Stage-2 response, while abciximab delayed and reduced the magnitude change of Stage-2. In the presence of collagen inhibitor, Stage-2 response was similar to that of control but was delayed by an additional 20 min. The obtained results, supported by epifluorescence and complementary SEM studies, demonstrated the selective sensitivity of TSM electromechanical biosensors to monitor platelet function and inhibition, particularly aggregation.     

Comparative toxic and sublethal effects of fluvalinate on two-spotted spider mite and European red mite

Four fluvalinate formulations differed in their residual toxicity to female two-spotted spider mite (TSM), Tetranychus urticae adults; the emulsifiable concentrate (EC) was the most toxic. In contrast, there was little difference in toxicity between the formulations with the European red mite (ERM) Panonychus ulmi with the exception of the EC formulation which was the least toxic. Fluvalinate 2F caused minimal (<10%) TSM and ERM egg mortality. Fluvalinate 2F was more toxic and caused greater larval dispersal for the TSM compared to the ERM at the field concentration and below. The toxicity of fluvalinate 2F to TSM and ERM protonymphs, deutonymphs and adults was low, approximately <20% at field concentration. Dispersal was the main response to fluvalinate and this was positively correlated with increasing concentration. The combined mortality and dispersal LC₅₀ was five times lower for ERM protonymphs and adults, but 11 times higher for ERM deutonymphs compared to equivalent TSM life stages. Fluvalinate 2F reduced TSM development from the protonymph and deutonymph stages to a greater extent compared to the ERM. The mortality response to fluvalinate 2F was unaffected by host type (peach or apple) for the TSM whereas ERM mortality was higher on apple compared to peach. TSM dispersal was higher from apple compared to peach whereas ERM dispersal was similar on both host types. Oviposition by both mite species was lower on apple than peach leaves. A 1 h exposure to fluvalinate 2F reduced ERM oviposition for 12 days.     

Effects of tracheal irritation and hypercapnia on tracheal smooth muscle in humans

Both hypercapnia and tracheal irritation are known to constrict the airways in animals. To see whether similar responses occur in humans, we investigated tracheal smooth muscle (TSM) responses to hypercapnia and tracheal irritation with water in 14 paralyzed and anesthetized humans. TSM tone was monitored by measuring the pressure in the saline-filled cuff of the endotracheal tube. Although, tracheal irritation caused TSM constriction in 10 of 14 patients, 4 patients showed no TSM response. Administration of intravenous atropine attenuated the TSM constriction response. Hypercapnia did not cause any change in TSM tone in any of the 14 patients. These results indicate that in paralyzed and anesthetized humans, there exist interindividual differences in the TSM responses to tracheal irritation and that hypercapnia cannot be an effective stimulus for the TSM constriction.     

Unravelling the effects of elevated temperature on the physiological energetics of Bellamya bengalensis

Temperature is one of the key environmental factors affecting the eco-physiological responses of living organisms and is considered one of the utmost crucial factors in shaping the fundamental niche of a species. The purpose of the present study is to delineate the physiological response and changes in energy allocation strategy of Bellamya bengalensis, a freshwater gastropod in the anticipated summer elevated temperature in the future by measuring the growth, body conditions (change in total weight, change in organ to flesh weight ratio), physiological energetics (ingestion rate, absorption rate, respiration rate, excretion rate and Scope for Growth) and thermal performance, Arrhenius breakpoint temperature (ABT), thermal critical maxima (CTmax), warming tolerance (WT) as well as thermal safety margin (TSM) through a mesocosm experiment. We exposed the animals to three different temperatures, 25 °C (average habitat temperature for this animal) and elevated temperatures 30 °C, 35 °C for 30 days and changes in energy budget were measured twice (on 15th and 30th day). Significant changes were observed in body conditions as well as physiological energetics. The total body weight as well as the organ/flesh weight ratio, ingestion followed by absorption rate decreased whereas, respiration and excretion rate increased with elevated temperature treatments resulting in a negative Scope for Growth in adverse conditions. Though no profound impact was found on ABT/CTmax, the peak of thermal curve was considerably declined for animals that were reared in higher temperature treatments. Our data reflects that thermal stress greatly impact the physiological functioning and growth patterns of B. bengalensis which might jeopardize the freshwater ecosystem functioning in future climate change scenario.     

SUMO‐modified insulin‐like growth factor 1 receptor (IGF‐1R) increases cell cycle progression and cell proliferation

Increasing number of studies have shown nuclear localization of the insulin‐like growth factor 1 receptor (nIGF‐1R) in tumor cells and its links to adverse clinical outcome in various cancers. Any obvious cell physiological roles of nIGF‐1R have, however, still not been disclosed. Previously, we reported that IGF‐1R translocates to cell nucleus and modulates gene expression by binding to enhancers, provided that the receptor is SUMOylated. In this study, we constructed stable transfectants of wild type IGF1R (WT) and triple‐SUMO‐site‐mutated IGF1R (TSM) using igf1r knockout mouse fibroblasts (R‐). Cell clones (R‐WT and R‐TSM) expressing equal amounts of IGF‐1R were selected for experiments. Phosphorylation of IGF‐1R, Akt, and Erk upon IGF‐1 stimulation was equal in R‐WT and R‐TSM. WT was confirmed to enter nuclei. TSM did also undergo nuclear translocation, although to a lesser extent. This may be explained by that TSM heterodimerizes with insulin receptor, which is known to translocate to cell nuclei. R‐WT proliferated substantially faster than R‐TSM, which did not differ significantly from the empty vector control. Upon IGF‐1 stimulation G1‐S‐phase progression of R‐WT increased from 12 to 38%, compared to 13 to 20% of R‐TSM. The G1‐S progression of R‐WT correlated with increased expression of cyclin D1, A, and CDK2, as well as downregulation of p27. This suggests that SUMO‐IGF‐1R affects upstream mechanisms that control and coordinate expression of cell cycle regulators. Further studies to identify such SUMO‐IGF‐1R dependent mechanisms seem important.     

Differential response of dog and pig tracheal smooth muscle to the acetylcholinesterase inhibitor soman

The effect of the acetylcholinesterase inhibitor soman on tracheal smooth muscle (TSM) from the dog and pig was studied. In response to soman, tracheal ring preparations contract more and the resting tension for TSM preparations is higher for the dog compared with the pig. Tension induced by electrical field stimulation (EFS) and the half-time of EFS-train induced contractions have a similar dependence on soman exposure in both dog and pig TSM. These results suggest that the basal acetylcholine secretion or leakage within the TSM nerve terminal is probably higher for the dog compared with the pig.     

Effects of constant or variable thermal rearing condition: a behavioural approach using an automated tracking system in Blaberus craniifer

An automatic tracking device was used to investigate exploratory behaviour of the cockroach, Blaberus craniifer, with regard to the influence of rearing temperature. Two groups were tested, one under variable rearing temperature, the other under constant rearing temperature. Rearing temperature influenced both the rate of decrease of linear speed and the spatial occupation of an open arena. Linear speed decreased with time and was higher during the first 10 minutes of recording for the animals reared under constant temperature conditions. The decrease fitted a linear model for the variable temperature group, whereas it was steeper than exponential for the constant temperature group. Frequency of occupancy in the peripheral zone of the arena was not affected by time and was always higher for the animals of the constant condition group. The results are interpreted as a function of the different thermal rearing conditions.     

Artificial tertiary motifs stabilize trans-cleaving hammerhead ribozymes under conditions of submillimolar divalent ions and high temperatures

Tertiary stabilizing motifs (TSMs) between terminal loops or internal bulges facilitate folding of natural hammerhead ribozymes (hRz) under physiological conditions. However, both substrate and enzyme strands contribute nucleotides to the TSMs of trans-cleaving hRz, complicating the design of hRz that exploit TSMs to target specific mRNA. To overcome this limitation, we used SELEX to identify new, artificial TSMs that are less sensitive to sequence context. Nucleotides in loop II or in a bulge within the ribozyme strand of stem I were randomized, while the interaction partner was held constant. All nucleotides of the substrate pair with the ribozyme, minimizing their possible recruitment into the TSM, as such recruitment could constrain choice of candidate target sequences. Six cycles of selection identified cis-acting ribozymes that were active in 100 microM MgCl2. The selected motifs partially recapitulate TSMs found in natural hRz, suggesting that the natural motifs are close to optimal for their respective contexts. Ribozyme "RzB" showed enhanced thermal stability by retaining trans-cleavage activity at 80 degrees C in 10 mM MgCl2 and at 70 degrees C in 2 mM MgCl2. A variant of ribozyme "RzB" with a continuously paired stem 1 rapidly lost activity as temperature was increased. The selected motifs are modular, in that they permit trans-cleavage of several substrates in submillimolar MgCl2, including two substrates derived from the U5 genomic region of HIV-1. The new, artificial tertiary stabilized hRz are thus nearly independent of sequence context and enable for the first time the use of highly active hRz targeting almost any mRNA at physiologically relevant magnesium concentrations.     

基于GOCI影像的湖泊悬浮物浓度分类反演

悬浮物直接影响到光在水体中的传播,进而影响着水生生态环境,最终决定了湖泊的初级生产力。传统的遥感反演估算模型大多是针对某一湖区进行统一建模,忽视了不同区域水体光学性质的复杂差异性,并且传统的传感器时间分辨率和空间分辨率受到一定限制。针对太湖、巢湖、滇池、洞庭湖4个湖区利用两步聚类法将高光谱模拟到GOCI影像上的波段进行分类,将水体类型分为三类,第一类水体为悬浮物主导的水体,第二类水体为悬浮物和叶绿素a共同主导的水体,第三类水体为叶绿素a主导的水体。针对不同类型水体的光学特征,分别构建了悬浮物浓度反演模型,结果表明第一类水体可以利用B7/B4,第二和第三类水体可以利用B7/(B8+B4)作为波段组合因子对悬浮物浓度进行模型构建。精度验证结果表明,分类建模后第一类和第三类水体悬浮物浓度估算精度都得到了较明显提高,第一类水体RMSE降低了9.19mg/L,MAPE降低了3%,第三类水体RMSE降低了5.63 mg/L,MAPE降低了13.97%,第二类水体精度稍有降低。最后将反演模型应用于2013年5月13日的GOCI影像,可知整体而言太湖西南部地区悬浮物浓度较高,东北部地区悬浮物浓度较低,并且从9:00到15:00,太湖南部悬浮物浓度较高的区域在逐渐缩小。     

Thermal niche variation among individuals of the poison frog,Oophaga pumilio,in forest and converted habitats

The conversion of natural habitats to human land uses often increases local temperatures, creating novel thermal environments for species. The variable responses of ectotherms to habitat conversion, where some species decline while others persist, can partly be explained by variation among species in their thermal niches. However, few studies have examined thermal niche variation within species and across forest‐land use ecotones, information that could provide clues about the capacity of species to adapt to changing temperatures. Here, we quantify individual‐level variation in thermal traits of the tropical poison frog, Oophaga pumilio, in thermally contrasting habitats. Specifically, we examined local environmental temperatures, field body temperatures (T_b), preferred body temperatures (T_pref), critical thermal maxima (CT_max), and thermal safety margins (TSM) of individuals from warm, converted habitats and cool forests. We found that frogs from converted habitats exhibited greater mean T_b and T_pref than those from forests. In contrast, CT_max and TSM did not differ significantly between habitats. However, CT_max did increase moderately with increasing T_b, suggesting that changes in CT_max may be driven by microscale temperature exposure within habitats rather than by mean habitat conditions. Although O. pumilio exhibited moderate divergence in T_pref, CT_max appears to be less labile between habitats, possibly due to the ability of frogs in converted habitats to maintain their T_b below air temperatures that reach or exceed CT_max. Selective pressures on thermal tolerances may increase, however, with the loss of buffering microhabitats and increased frequency of extreme temperatures expected under future habitat degradation and climate warming. Abstract in Spanish is available with online material.     

A critical assessment of the topomer search model of protein folding using a continuum explicit-chain model with extensive conformational sampling

Recently, a series of closely related theoretical constructs termed the "topomer search model" (TSM) has been proposed for the folding mechanism of small, single-domain proteins. A basic assumption of the proposed scenarios is that the rate-limiting step in folding is an essentially unbiased, diffusive search for a conformational state called the native topomer defined by an overall native-like topological pattern. Successes in correlating TSM-predicted folding rates with that of real proteins have been interpreted as experimental support for the model. To better delineate the physics entailed, key TSM concepts are examined here using extensive Langevin dynamics simulations of continuum C(alpha) chain models. The theoretical native topomers of four experimentally well-studied two-state proteins are characterized. Consistent with the TSM perspective, we found that the sizes of the native topomers increase with experimental folding rate. However, a careful determination of the corresponding probabilities that the native topomers are populated during a random search fails to reproduce the previously predicted folding rates. Instead, our results indicate that an unbiased TSM search for the native topomer amounts to a Levinthal-like process that would take an impossibly long average time to complete. Furthermore, intraprotein contacts in all four native topomers considered exhibit no apparent correlation with the experimental phi-values determined from the folding kinetics of these proteins. Thus, the present findings suggest that certain basic, generic yet essential energetic features in protein folding are not accounted for by TSM scenarios to date.     

Prey capture success of Sandwich Terns Sterna sandvicensis varies non‐linearly with water transparency

Human impacts on water transparency may affect plunge‐diving seabirds. We studied prey capture success of Sandwich Terns Sterna sandvicensis as a function of six environmental variables during the breeding season. We observed diving terns in the south eastern North Sea and found a non‐linear optimum curve for the relationship between Secchi transparency and prey capture probability. High capture probability was found at 1.5–2.0 m, with an optimum of 63% at 1.74 m. At a minimum transparency of 0.4 m and at a maximum transparency of 3.2 m, capture probabilities were about halved. Conversion of transparency to total suspended matter (TSM) concentration showed that the optimum concentration for foraging terns would be 5–10 mg/L under mean summer conditions for chlorophyll‐a. However, the summer‐averaged TSM concentrations in the nearshore Dutch coastal waters range between 10 and 30 mg/L, which implies that foraging Terns in the breeding season do not encounter optimal foraging conditions. The Full Plunge Dive, with which the largest diving depth can be achieved, was applied dominantly in clear water, while the Partial Plunge Dive and Contact Dip were applied more frequently in turbid water, thus showing that Sandwich Terns adjust their diving technique in response to water transparency.     

Induction of cytochrome P-450 by triterpensaponins in Vietnamese ginseng

It was found that rat liver cytochrome P-450 is induced by the Vietnamese ginseng triterpensaponines mixture (TSM) as well as by K5VN Panaxozides-11 (VP-11) purified from this mixture. Addition of TSM and VP-11 accelerates benz(alpha)pyrene and aminopyrine hydroxylation and increases the content of cytochrome P-450 isoforms with Mr of 57 kDa and 54 kDa in rat liver microsomes. Since VP-11 accounts for about 50% of TSM, the results obtained suggest that the microsomal monooxygenase system induction is caused by this triterpensaponine. Induction by TSM and VP-11 was compared to that by phenobarbital (PB) and 3-methylcholanthrene (MC). It was shown that according to their inductive action TSM and VP-11 belong neither to the PB- nor to the MC-type. Cytochrome P-450 induction may play an important role in the triterpensaponine action on the organism, because this enzyme participates in the metabolism of such endogenous compounds as prostaglandins, steroid hormones, cholesterol, etc.     

Comparative Estimation of Genetic Diversity in Population Studies using Molecular Sampling and Traditional Sampling Methods

Entomopathogenic nematodes (EPN) are efficient biological pest control agents. Population genetics studies on EPN are seldom known. Therefore, it is of interest to evaluate the significance of molecular sampling method (MSM) for accuracy, time needed, and cost effectiveness over traditional sampling method (TSM). The study was conducted at the Mohican Hills golf course at the state of Ohio where the EPN H. bacteriophora has been monitored for 18 years. The nematode population occupies an area of approximately 3700 m² with density range from 0.25-2 per gram soil. Genetic diversity of EPN was studied by molecular sampling method (MSM) and traditional sampling method (TSM) using the mitochondrial gene pcox1. The MSM picked 88% in compared to TSM with only 30% of sequenced cox 1 gene. All studied genetic polymorphism measures (sequence and haplotype) showed high levels of genetic diversity of MSM over TSM. MSM minimizes the chance of mitochondrial genes amplification from non target organisms (insect or other contaminating microorganisms). Moreover, it allows the sampling of more individuals with a reliable and credible representative sample size. Thus, we show that MSM supersedes TSM in labour intensity, time consumption and requirement of no special experience and efficiency.     

Heat tolerance variation reveals vulnerability of tropical herbivore–parasitoid interactions to climate change

Assessing the heat tolerance (CTmax) of organisms is central to understand the impact of climate change on biodiversity. While both environment and evolutionary history affect CTmax, it remains unclear how these factors and their interplay influence ecological interactions, communities and ecosystems under climate change. We collected and reared caterpillars and parasitoids from canopy and ground layers in different seasons in a tropical rainforest. We tested the CTmax and Thermal Safety Margins (TSM) of these food webs with implications for how species interactions could shift under climate change. We identified strong influence of phylogeny in herbivore–parasitoid community heat tolerance. The TSM of all insects were narrower in the canopy and parasitoids had lower heat tolerance compared to their hosts. Our CTmax-based simulation showed higher herbivore–parasitoid food web instability under climate change than previously assumed, highlighting the vulnerability of parasitoids and related herbivore control in tropical rainforests, particularly in the forest canopy.     

Monitoring cell adhesion by using thickness shear mode acoustic wave sensors

The thickness shear mode (TSM) acoustic wave sensor attached with living cells has been shown to be an effective functional biosensing device to monitor the process of cell adhesion to a surface in real time. In this study, a multilayer sensor model that includes a quartz substrate, a cell-substrate interfacial layer and a cell layer was constructed based on the state of cell adhesion to the substrate. The dynamic process of cell adhesions as a function of cell seeding densities was monitored using the acoustic wave sensor. The mechanisms that are responsible for the frequency and resistance change are discussed according to the predictions of the acoustic wave sensor model. In addition, knowing that the actin cytoskeleton is important for cell adhesion, we investigated the motional resistance change caused by the disruption of actin cytoskeleton induced by fungal toxin cytochalasin D in the human skin fibroblasts. The results indicate that resistance changes are related to the disruption of actin cytoskeleton and dosage-dependent.     

NO responsiveness in pulmonary artery and airway smooth muscle: the role of cGMP regulation

The purpose of this study was to assess intrinsic smooth muscle mechanisms contributing to greater nitric oxide (NO) responsiveness in pulmonary vascular vs. airway smooth muscle. Canine pulmonary artery smooth muscle (PASM) and tracheal smooth muscle (TSM) strips were used to perform concentration response studies to an NO donor, (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate (DETA-NO). PASM exhibited a greater NO responsiveness whether PASM and TSM were contracted with receptor agonists, phenylephrine and acetylcholine, respectively, or with KCl. The >10-fold difference in NO sensitivity in PASM was observed with both submaximal and maximal contractions. This difference in NO responsiveness was not due to differences in endothelial or epithelial barriers, since these were removed, nor was it due to the presence of cGMP-independent NO-mediated relaxation in either tissue. At equal concentrations of NO, the intracellular cGMP concentration ([cGMP]i) was also greater in PASM than in TSM. Phosphodiesterase (PDE) inhibition using isobutylmethylxanthine indicated that the greater [cGMP]i in PASM was not due to greater PDE activity in TSM. Expression of soluble guanylate cyclase (sGC) subunit mRNA (2 +/- 0.2 and 1.3 +/- 0.2 attomol/microg total RNA, respectively) and protein (47.4 +/- 2 and 27.8 +/- 3.9 ng/mg soluble homogenate protein, respectively) was greater in PASM than in TSM. sGCalpha1 and sGCbeta1 mRNA expression was equal in PASM but was significantly different in TSM, suggesting independent regulation of their expression. An intrinsic smooth muscle mechanism accounting for greater NO responsiveness in PASM vs. TSM is greater sGC activity.     

Hypermutagenesis in mutA cells is mediated by mistranslational corruption of polymerase, and is accompanied by replication fork collapse

Elevated mistranslation induces a mutator response termed translational stress‐induced mutagenesis (TSM) that is mediated by an unidentified modification of DNA polymerase III. Here we address two questions: (i) does TSM result from direct polymerase corruption, or from an indirect pathway triggered by increased protein turnover? (ii) Why are homologous recombination functions required for the expression of TSM under certain conditions, but not others? We show that replication of bacteriophage T4 in cells expressing the mutA allele of the glyV tRNA gene (Asp→Gly mistranslation), leads to both increased mutagenesis, and to an altered mutational specificity, results that strongly support mistranslational corruption of DNA polymerase. We also show that expression of mutA, which confers a recA‐dependent mutator phenotype, leads to increased lambdoid prophage induction (selectable in vivo expression technology assay), suggesting that replication fork collapse occurs more frequently in mutA cells relative to control cells. No such increase in prophage induction is seen in cells expressing alaV^Glu tRNA (Glu→Ala mistranslation), in which the mutator phenotype is recA‐independent. We propose that replication fork collapse accompanies episodic hypermutagenic replication cycles in mutA cells, requiring homologous recombination functions for fork recovery, and therefore, for mutation recovery. These findings highlight hitherto under‐appreciated links among translation, replication and recombination, and suggest that translational fidelity, which is affected by genetic and environmental signals, is a key modulator of replication fidelity.     

中国根层与表层土壤水分关系分析

根层与表层土壤水分的关系, 是由较易获取的表层土壤水分信息去探讨较难获取的深层土壤水分信息的重要桥梁。已有的根层与表层土壤水分关系(简称根表关系)大都基于一种作物或一种生态系统。该文根据我国生态系统研究网络, 包括森林、草地、农田、荒漠和沼泽生态系统的31个站点109个观测场2006年全年3437对根层和表层土壤水分数据, 研究了根表关系以及生态系统、土壤质地、湿润度、植被、土壤厚度和土壤水分量级对根表关系的影响。研究发现, 表层和根层土壤水分存在着线性关系。森林和沼泽的根层与表层土壤水分相关程度较高, 无论是率定段还是校核段, 其决定系数(R²)均大于0.79。农田和草地生态系统的率定段相关性较好, R ²均大于0.80, 校核段相关性稍弱, R ²分别为0.70和0.50。荒漠生态系统的相关关系最弱, 率定段的R²为0.62, 校核段的R²为0.49。土壤质地和生态系统因素对根表关系的影响较为一致。半湿润带、半干旱带和干旱带的根表关系空间分异性最强; 十分湿润带的根表关系与壤土和森林生态系统的根表关系相对应。湿润带内部的根表关系较为一致。将植被对根表关系的影响分为4类, 前两类为根表关系微弱的植被, 由植被本身或者植被以外的地域因素导致, 不适合用根表关系去由表层推算根层土壤水分; 后两类为根表关系良好植被, 区别为服从和不服从关系总线, 可分别用各自的根表关系或者关系总线从表层土壤水分获取根层土壤水分。表层土壤水分与0-20、0-30、……、0-100 cm土层的土壤水分均分别具有较好的相关关系, 但二者的相关性随土层厚度的增加而降低。不过, 即使是土层厚度抵及100 cm, R ²仍能维持在0.79。通过将土壤水分分别除以所有观测数据的最大值(“标甲”法)和各个生态系统数据的最大值(“标乙”法), 发现根表关系不受土壤水分量级本身的影响。