Clinorotation Effect on Response of Cress Leaves to Red and Far-Red Light

Responses of cress (Lepidium sativum L.) seedling leaves to separate and simultaneous illumination with red (660 nm) and far-red (735 nm) light were studied under fast clinorotation (50 rpm) and usual gravity (1g) conditions. The monochromatic light emitting diodes (LEDs) have been used for illumination of seedlings from above. The growth and spatial orientation of leaves and the location of presumable gravisensors in petioles were analysed. Clinorotation in the dark promoted the radial expansion of leaf lamina and unfolding of leaves. It was shown that clinorotation in red light inhibited significantly the elongation of petioles as compared with that under the action of gravity force. Simultaneous red and far-red illumination promoted the growth of petioles under clinorotation, but did not affect the orientation of laminas, which remained the same as of the 1-g control ones. Red light, applied separately and simultaneously with far-red light, guided the bending of laminas as well as the unfolding of leaf petioles in both usual and clinorotation conditions. Histological and cytological analyses of petioles revealed the presence of movable amyloplasts in endodermic cells in proximal region of petioles. Comparison of intracellular positioning of amyloplasts in petioles of leaves grown under clinorotation and the action of gravity allows a presumption that these plastids may be identified as gravisensors of garden cress leaves. The obtained data imply that clinorotation and exposition to monochromatic red light or simultaneous illumination by red and far-red light affect the elongation of petioles of cress seedling leaves. Spectral components guide the unfolding of laminas in a gravity-independent manner.     

Analysis of Statoliths Displacement in <Emphasis Type="Italic">Chara</Emphasis> Rhizoids for Validating the Microgravity-Simulation Quality of Clinorotation Modes

In single-celled rhizoids of the green algae Chara, positively gravitropic growth is governed by statoliths kept in a dynamically stable position 10–25 μ m above the cell tip by a complex interaction of gravity and actomyosin forces. Any deviation of the tube-like cells from the tip-downward orientation causes statoliths to sediment onto the gravisensitive subapical cell flank which initiates a gravitropic curvature response. Microgravity experiments have shown that abolishing the net tip-directed gravity force results in an actomyosin-mediated axial displacement of statoliths away from the cell tip. The present study was performed to critically assess the quality of microgravity simulation provided by different operational modes of a Random Positioning Machine (RPM) running with one axis (2D mode) or two axes (3D mode) and different rotational speeds (2D), speed ranges and directions (3D). The effects of 2D and 3D rotation were compared with data from experiments in real microgravity conditions (MAXUS sounding rocket missions). Rotational speeds in the range of 60–85 rpm in 2D and 3D modes resulted in a similar kinetics of statolith displacement as compared to real microgravity data, while slower clinorotation (2–11 rpm) caused a reduced axial displacement and a more dispersed arrangement of statoliths closer to the cell tip. Increasing the complexity of rotation by adding a second rotation axis in case of 3D clinorotation did not increase the quality of microgravity simulation, however, increased side effects such as the level of vibrations resulting in a more dispersed arrangement of statoliths. In conclusion, fast 2D clinorotation provides the most appropriate microgravity simulation for investigating the graviperception mechanism in Chara rhizoids, whereas slower clinorotation speeds and rotating samples around two axes do not improve the quality of microgravity simulation.     

From ROOTS to GRAVI-1: Twenty Five Years for Understanding How Plants Sense Gravity

In the 1970s, when I started to work on gravitropism at the University Pierre and Marie Curie, Paris), it was well known that statocytes contain voluminous amyloplasts (statoliths) that sediment under the influence of gravity. The role of these organelles in gravisensing was strongly disputed. In 1974, I attended a session of a meeting on gravitropism in Würzburg, where I presented results that supported the involvement of statoliths in the perception of gravity. This meeting had a strong impact on my research, since at that time the Council of Europe was looking for people interested in performing experiments in Space. Our first experiment (ROOTS) was carried out in the Biorack Facility (ESA) in the frame of the Spacelab D1 mission (1985). We had a very efficient help from CNES which developed a very fine hardware to grow lentil seedlings and to chemically fix them at the end of the experiment. The results obtained were surprising since we observed that in microgravity the statoliths were located at one pole of the statocyte and not distributed at random as it was expected. The goal of the following experiment (Spacelab IML-1 mission, 1992) was to determine the threshold stimulation time at 1 ×g (created by centrifugation). It was estimated at 25 s. In the frame of the SMM/03 and SMM/06 missions (1996, 1997), we proved that the statoliths are attached to actin filaments by motor proteins (myosin) that make these organelles move in one preferential direction in microgravity. The analysis of gravisensitivity with clinostats incited us to compare gravisensitivity of lentil roots grown in microgravity or on a 1 × g centrifuge (SMM05 mission, 1997). It was found that the latter were less sensitive than the former. We showed that this was due to the fact that the statoliths are not distributed in the same way in both cases (microgravity or 1 × g centrifuge). All these studies led us to propose a mode of gravity sensing by plants in which elements of the cytoskeleton and stretch activated ion channels are involved. The last experiment (GRAVI-1) which has been carried out (in ISS with the EMCS facility, 2007) dealt with the threshold acceleration that is perceived by roots.     

Effects of Clinorotation on Growth and Chlorophyll Content of Rice Seeds

Microgravity, as a different environment, has been shown to affect plant growth and development (Sievers et al. 1996; Sack 1997). In the present study, effects of slow clinorotation (2 rpm) on growth and chlorophyll content in rice (variety: PRH-10) seedlings were investigated. Rice seeds were clinorotated continuously for 3, 5 and 7 days under ambient conditions. Root and shoot lengths and weights of rice seedlings were measured on the third, fifth and seventh day. Chlorophyll was extracted using N, N-Dimethylformamide (DMF). Absorption and fluorescence spectra of chlorophyll were recorded. Chlorophyll a, chlorophyll b and total chlorophyll contents were calculated from absorption spectra using Arnon’s method. Results showed an increase in root and shoot lengths in clinorotated samples. Similar results were obtained for root and shoot weights. Absorption spectra of chlorophyll showed no shift in the absorption peaks. Chlorophyll content was increased in clinorotated samples as compared to the controls. Interestingly, the difference between chlorophyll content in control and clinorotated samples decreased as the number of days of clinorotation increased. Chlorophyll a/b ratio was lowered in clinorotated samples as compared to the controls. These results suggest that slow clinorotation (2 rpm) affects plant growth and chlorophyll content in rice seedlings.     

Tubulin Cytoskeleton in Arabidopsis thaliana Root Cells Under Clinorotation

The effect of horizontal clinorotation (2 rpm) on the spatial organization of cortical microtubules (CMTs) in the living Arabidopsis root cells has been investigated. MTs were visualized by using a stably transformed line of transgenic Arabidopsis thaliana expressing GFP-MAP4 fusion protein. To monitor total changes in MT dynamics, the sensitivity to MT-inhibitor 5 μM/l oryzalin was used as an indicator. It was shown that CMTs in the distal elongation zone (DEZ) of clinorotated 5 day old seedlings exhibited a higher degree of disorder and also they were more sensitive to oryzalin. We suggest that rearrangement of CMTs in DEZ cells under clinorotation is caused by fast rate of MT dynamic turnover.     

Actin Microfilament Organization in the Transition Zone of Arabidopsis-ABD2-GFP Roots Under Clinorotation

Seedlings of Arabidopsis thaliana-ABD2-GFP were grown under slow clinorotation (2?rpm) and treated with actin and tubulin disrupting drugs in order to characterize the role of actin microfilaments in cell growth and gravisensing. Changes in microfilament organization and cell parameters have shown that the transition root zone (TZ) is rather sensitive to microfilament disruption in control plants. It is assumed that under clinorotation, organization of actin cytoskeleton in the TZ is coordinated in a different way than in the control. Organization of microfilaments depends upon organization of microtubules and clinorotation does not influence this interrelation significantly.     

Distribution of Calcium Ions in Cells of the Root Distal Elongation Zone Under Clinorotation

Currently, calcium ions are known to play a crucial role in the vital activity of plant cells and in stimulus–response coupling for many environmental signals, altered gravity included. The available data on changes in Ca^2?+? distribution and concentration in the cells of different organisms influenced by altered gravity allow to suggest that microgravity affects the calcium messenger system, and provide new insight for the understanding of calcium-and gravity-dependent cellular processes. We have studied with confocal microscopy the distribution and relative content of calcium ions in the Beta vulgaris root distal elongation zone cells grown under slow horizontal clinorotation, reproducing one of the microgravity particularities, namely the absence of an orienting action of the gravity vector, compared to control conditions. We demonstrate that Ca^2?+? relative content is 1.3 times higher in the roots of seedlings grown upwards and 1.2 times higher in the seedlings grown downwards compared to the control. Based on obtained data, taking into account the specific physiological properties of cells in the distal elongation zone, it is supposed that, under clinorotation, enhanced Ca^2?+? relative content affects Ca^2?+?-dependent cytoskeleton reorganization involved in cell gravisensing in altered gravity.     

The stress protein level under clinorotation in context of the seedling developmental program and the stress response

Heat-shock proteins (HSP70 and HSP90) are present in plant cells under the normal growth conditions. At the same time, a variety of environmental disruptions results in their rapid synthesis as a substantial part of adaptation. HSP amounts can be indicative of a cellular stress level. Altered gravity (clinorotation) is unnatural for plants, so it may be a kind of stress. The aim of this study was to analyze the influence of horizontal clinorotation on the HSP70 and HSP90 level during seedling development. Pea (Pisum sativum L.) seedlings grown for 3 days from seed imbibitions in stationary control and under slow clinorotation (2 rpm) are used for this investigation. Western blot analysis indicated that HSP70 and HSP90 were abundant in the embryos of dry seeds and their amount decreased significantly during seed germination. But under horizontal clinorotation, their level in seedlings remained higher compared to the control. Furthermore, a comparison of the influence of horizontal and vertical clinorotation on the HSP level was carried out. On the ELISA data, HSP70 and HSP90 amounts in the 3-day old seedlings were higher after horizontal clinorotation than after vertical. The obtained data show an increased HSP70 and HSP90 level in pea seedlings under clinorotation. Both, rotation and change in the cell position relatively to a gravity vector affect the HSP level.     

Effects of Altered Gravity on the Cytoskeleton of Neonatal Rat Cardiocytes

As an intracellular load-bearing structure, the cytoskeleton is hypothesized to play a crucial role in gravity perception of the cell. Recent data show that the cytoskeleton, which includes actin microfilaments and microtubules, is involved in modulating both the electrical and the mechanical activities of the myocardium. The present study employed observation and quantified analyses of fluorescent images of cardiocytes under different gravity conditions. In acute gravitational change (micro- and hypergravity) induced by parabolic flight, we found disassembly of microtubules but enhanced polymerization of microfilaments, with rearrangement from G-actin to F-actin. In ground-based experiments, exposure of cardiocytes to 2×g hypergravity (centrifugation) led to increased width and number of actin fibers from 2 to 48 h, while microtubules showed no significant changes except polarization at 24 and 48 h. In contrast, exposure of cardiocytes to clinorotation led to disassembly of microtubules from 1 to 48 h, while microfilaments showed no significant changes except redistribution, which was accompanied by rounding of the cells (48 h). We assume that the sensitivity of microfilaments to hypergravity and that of microtubules to microgravity might contribute to the specific cytoskeletal changes observed in parabolic flight. These findings indicate different sensitivity and responses of microfilaments and microtubules to different gravitational changes, which might be part of functional adaptations of the cardiocytes to altered gravitational environments.     

Comparative studies on gravisensitive protists on ground (2D and 3D clinostats) and in microgravity

In order to prepare and support space experiments, 2D and 3D clinostats are widely applied to study the influence of simulated weightlessness on biological systems. In order to evaluate the results a comparison between the data obtained in simulation experiments and in real microgravity is necessary. We are currently analyzing the gravity-dependent behavior of the protists Paramecium biaurelia (ciliate) and Euglena gracilis (photosynthetic flagellate) on these different experimental platforms. So far, first results are presented concerning the behaviour of Euglena on a 2D fast rotating clinostat and a 3D clinostat as well as under real microgravity conditions (TEXUS sounding rocket flight), of Paramecium on a 2D clinostat and in microgravity. Our data show similar results during 2D and 3D clinorotation compared to real microgravity with respect to loss of orientation (gravitaxis) of Paramecium and Euglena and a decrease of linearity of the cell tracks of Euglena. However, the increase of the mean swimming velocities, especially during 3D clinorotation (Euglena) and 2D clinorotation of Paramecium might indicate a persisting mechanostimulation of the cells. Further studies including long-term 2D and 3D clinostat exposition will enable us to demonstrate the qualification of the applied simulation methods.     

Modeling the Kinetics of Root Gravireaction

The known “sun-flower equation” (SFE), which was originally proposed to model root circumnutating, was used to describe the simplest tip root graviresponse. Two forms of the SFE (integro-differential and differential-delayed) were solved, analyzed and compared with each other. The numerical solutions of these equations were found to be matching with arbitrary accuracy. The analysis of the solutions focused on time-lag effects on the kinetics of tip root bending. The results of the modeling are in good correlation with an experiment at the initial stages of root tips graviresponse. Further development of the model calls for its systematic comparison with some specially designed experiments, which would include measuring the kinetics of root tip bending before gravistimulation over the period of time longer than the time lag.     

Cress seed mucilage based buccal mucoadhesive gel of venlafaxine: in vivo,in vitro evaluation

Venlafaxine is a newer antipsychotic drug which shows first pass effect. Cress seed is also called as garden cress or green salad. This study examined the mechanical (gel strength, adhesiveness) and rheological properties of cress seed mucilage based gels that contain different ratios of carbopol 934 P (0.5–1.5%). In addition, diffusion of venlafaxine from gel formulations was evaluated. The selected formulation was further analyzed for pharmacokinetic parameters in rabbits. All formulations exhibited pseudoplastic flow with thixotropy. Formulation F5 showed the C_max of 24.19 ± 0.72 ng/ml by buccal route of administration and 17.98 ± 1.15 ng/ml by oral route of administration. The bioavailability of F5 by buccal route was 54.44% and that of by oral route was 39.60%. A combination of the cress seed mucilage and carbopol 934 P resulted in a prolonged and higher venlafaxine delivery by buccal route of administration.     

Notch root contraction as a fracture measurement

The value of displacement measurements in fracture toughness testing is discussed; in particular the measurement of notch root contraction. The mechanisms of plastic deformation at the notch tip are described, leading to a discussion of theoretical relationship between notch root contraction and crack opening displacement. Experimental work is described in which notched bend specimens of a low alloy steel were loaded to fracture. Each specimen was simultaneously instrumented with notch contraction, COD, and “clip-in” gauges. The equality between notch root contraction and COD was confirmed and observations are made on the relative value of the different measurements. Further observations on the distribution of contraction around the notch tip were made.     

Chlorophyll,Carotenoid and Anthocyanin Accumulation in Mung Bean Seedling Under Clinorotation

The accumulation of plant pigments in mung bean (Vigna radiata L.) seedlings was measured after clinorotation (2 rpm for 2-4 days), and compared to a stationary control. The pigments measured included chlorophyll and carotenoid in primary leaves, and the anthocyanin in seedlings. While significant changes in chlorophyll and carotenoid accumulation were not observed during the initial 2 to 4 days of cultivation, by day 4 the seedlings grown on the clinostat had lower levels of anthocyanin, compared to those in the control seedlings. To further detail the cause for the observed reduction in anthocyanin accumulation under altered gravity conditions, seedlings were grown in the presence of silver nitrate, a known ethylene inhibitor, for 4 days, since it is known ethylene has a negative impact on anthocyanin accumulation. Silver nitrate promoted anthocyanin accumulation in the clinostat seedlings, and as a result there was no significant difference between the control and clinostat seedlings in anthocyanin accumulation. The results suggest that slow clinorotation negatively impacts anthocyanin pigmentation in mung bean seedlings, with endogenous ethylene suspected to be involved in this.     

基于多模态展开的黏弹性材料复纵波波速反演方法

黏弹性材料广泛应用于水下航行器的降噪、减振或消声,精确获取其动力学参数的需求随着数值仿真技术发展而迅速提升.针对参数获取的声管测量方式,提出了一种使用复反射系数对黏弹性材料复纵波波速快速反演的方法.通过引入多模态展开方法,将复纵波波数超越方程的复平面搜根问题转化为展开系数系统矩阵特征值分解问题,推导出了三种典型背衬情况...     

Description of the kinetics of growth of a fatigue crack on the basis of the criterion of current crack tip opening displacement

We suggest a microprocessor-based data-processing and measuring system and a special device aimed at the determination of current values of the crack tip opening displacement. They are used to establish current values of the crack tip opening displacement at working frequencies of loading of the specimen and describe the kinetics of fatigue crack growth. A decrease in the loading frequency of the specimen from 15 to 0.1 Hz in the course of measuring the current values of the crack tip opening displacement does not affect the reliability of determination of the characteristics of cyclic crack-growth resistance. The reliability of the results is affected only by the inertia of the mass of the gauge case. Institute for Problems of Strength, National Academy of Sciences of Ukraine, Kiev, Ukraine. Translated from Problemy Prochnosti, No. 4, pp. 60–67, July–August, 1998.     

Fatigue cracks emanating from sharp notches in high-strength aluminium alloys: The effect of loading direction, kinking, notch geometry and microstructure

The effect of notch geometry on the propagation of fatigue cracks emanating from sharp V-shaped notches was investigated. To this purpose, an experimental campaign has been conducted on Al-7075–T651 specimens carrying notches with aperture angles of 45°, 90°, and 135°. In order to investigate the role of microstructure texture, specimens were extracted from the plates with the main axis either in the longitudinal rolling direction (L-samples) or in the transversal direction (T-samples), or 45° inclined with respect to both directions (LT-samples). The effect of stress amplitude was investigated by performing tests at two load levels. Three loading directions θ = 0°, 45° and 90° were considered. Some specimens experienced pure Mode I loading condition, whereas the remaining ones were subjected to combined Mode I and Mode II loading condition. The crack deflection induced by the variation in loading direction was determined by measuring the kinking angle. A linear elastic fracture mechanics approach was adopted for the analysis of experimental results. Stress intensity factors (SIF) of straight cracks were calculated using an appropriate weight function set up for studying inclined edge cracks emanating from sharp V-notches. On the contrary, a finite element model has been built up to derive the SIFs at the tip of the kinked cracks. The influence of K_II on the crack propagation was discussed on the basis of theoretical and semi-empirical models. It has been found that (i) the crack initiation at the notch root occurred in mixed mode conditions, (ii) a decreasing Mode II component with growing crack length was observed under initial loading direction θ₀ = 45° and θ₀ = 90°, (iii) a crack deflection was observed after 45° rotation of the initial loading direction; a good prediction of the kinking angle was obtained using the maximum tangential stress criterion, and (iv) a fairly good rationalization of all the collected crack growth rate data is obtained if the driving force for crack propagation is expressed in terms of K_I.     

Rocket seedling production on the international space station: Growth and nutritional properties

Producing sprouts directly during space missions may represent an interesting opportunity to offer high-quality fresh ready to eat food to the astronauts. The goal of this work was to compare, in terms of growth and nutritional quality, rocket (Eruca sativa Mill.) seedlings grown in the International Space Station during the ENEIDE mission with those grown in a ground-based experiment (in presence and absence of clinorotation). The rocket seedlings obtained from the space-experiment were thinner and more elongated than those obtained in the ground-based experiment. Cotyledons were often closed in the seedlings grown in the space experiment. Quantitative (germination, fresh and dry weight) and qualitative (glucose, fructose, sucrose and starch) traits of rocket seedling were negatively affected by micrograv-ity, especially those recorded on seedlings grown under real microgravity conditions The total chlorophyll, and carotenoids of seedlings obtained in the space experiment were strongly reduced in comparison to those obtained in the ground-based experiment (presence and absence of clinorotation). The results showed that it is possible to produce rocket seedlings in the ISS; however, further studies are needed to define the optimal environmental conditions for producing rocket seedlings with high nutritional value     

Development of a three-point bending fatigue testing methodology at 20 kHz frequency

A novel testing methodology on three-point bending fatigue test at 20 kHz has been developed based on ultrasonic vibration theory to determine the bending fatigue strength of structural materials in very high cycle regime. In this paper, the working feature of the testing machine has been introduced. On the basis of vibration theory analysis, the differential equation of longitudinal and flexural vibration were given, finite element model was introduced to design and calculate the amplifier, specimen and the connector in the testing system. Furthermore, to produce the shapes of the components necessary to achieve the required displacement and stress states. Longitudinal cyclic load was transferred to transverse vibration of specimen by the tip in the testing system. In order to understand the effect of the tip on the testing system, matching analysis between amplifier and tip in the bending vibration system has been discussed. Finally, a successful application of the methodology is demonstrated by the experimental results from TiAl alloy specimens subjected to three-point bending stress states.     

Crack resistance behavior of particulate reinforced composites at various test speeds and temperatures

In this study, the fracture behavior and characteristics of particulate-reinforced composite materials were evaluated by performing wedge splitting tests. The crack resistance of the materials was evaluated using the crack tip opening displacement and crack tip opening angle. The composites were tested under various temperatures and test speeds. The digital image correlation method was used to analyze the strain field at the crack tip. The fracture surface under test conditions was observed using a scanning electron microscope. The test results showed that the fracture energy increased with decreasing temperature, and the crack resistance increased with increasing test speed. The crack tip opening angle is divided into an unstable region and stable region. The critical crack tip opening angle can be defined as the fracture mechanics parameter measured in a stable region. The surface strain fields obtained by digital image correlation method are distributed in the range from 1.5 % to 4.5 % at the initiation of the crack. A crack grows with dewetting phenomenon at the temperature range from 60 °C to −40 °C, and the crack propagates with fracture of ammonium perchlorate oxidizer particles at the glass transition temperature of −70 °C.