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This work demonstrates that in photothermal experiments performed in frequency domain the heat losses due to convection and radiation should be taken into account at low frequencies for poor heat conductors. From a model-solution of the heat diffusion equation a dimensionless frequency dependent parameter M = ZH, with sample's thermal impedance Z and H the convection-radiation heat transfer coefficient, turns out to adequately quantify the importance of the effect of those heat losses. A straightforward photothermal infrared radiometry setup was designed to demonstrate the above hypothesis. Disc shaped samples of different test materials were heated by square wave modulated illumination at one of their surfaces at different frequencies using an amplitude modulated laser beam, and the temperature at the rear surfaces was monitored as a function of time using an infrared sensor. The frequency dependence of peak-to-peak values of the temperature signals was found to be consistent with the amplitude spectrum obtained by Fourier transforming the data. The frequency dependence of the peak-to-peak amplitude was compared with a theoretical model with and without taking convection and radiation induced heat losses (CRHL) into consideration. It is found that for poor heat conductors at low modulation frequencies the conventional model without CRHL does not fit well the experimental data, while the extended model leads to good agreement, resulting in reliable values for the thermal diffusivity. For the investigated samples, the contribution to the signal of thermal wave reflection at the back side of the sample turn out to have a minor effect on the signal spectrum.

Plants grown under iron (Fe)-deficient conditions induce a set of genes that enhance the efficiency of Fe uptake by the roots. In Arabidopsis (Arabidopsis thaliana), the central regulator of this response is the basic helix-loop-helix transcription factor FER-LIKE IRON DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (FIT). FIT activity is regulated by protein-protein interactions, which also serve to integrate external signals that stimulate and possibly inhibit Fe uptake. In the search of signaling components regulating FIT function, we identified ZINC FINGER OF ARABIDOPSIS THALIANA12 (ZAT12), an abiotic stress-induced transcription factor. ZAT12 interacted with FIT, dependent on the presence of the ethylene-responsive element-binding factor-associated amphiphilic repression motif. ZAT12 protein was found expressed in the root early differentiation zone, where its abundance was modulated in a root layer-specific manner. In the absence of ZAT12, FIT expression was upregulated, suggesting a negative effect of ZAT12 on Fe uptake. Consistently, zat12 loss-of-function mutants had higher Fe content than the wild type at sufficient Fe. We found that under Fe deficiency, hydrogen peroxide (H2O2) levels were enhanced in a FIT-dependent manner. FIT protein, in turn, was stabilized by H2O2 but only in the presence of ZAT12, showing that H2O2 serves as a signal for Fe deficiency responses. We propose that oxidative stress-induced ZAT12 functions as a negative regulator of Fe acquisition. A model where H2O2 mediates the negative regulation of plant responses to prolonged stress might be applicable to a variety of stress conditions.

A b s t r a c t. It is important to know the temperature changes in seeds that have been irradiated with laser light because this could have substantial practical and theoretical importance. Thus, the thermal effects of low intensity laser irradiation on seeds was studied, showing variation of temperature produced by laser light applied during 60 s on two maize seed genotypes, ‘Toluqueno’ and ‘Cacahuazintle’: crystalline and floury, respectively, under two different conditions: natural colour and dyed black, evaluating the temperature changes by a thermal camera. The optical absorption spectra and the non-radiative relaxation time of the seeds were obtained using photoacoustic spectroscopy. The results indicate that it is possible to produce temperature changes, detected by an infrared camera, in crystalline and floury seeds when they are irradiated with a laser beam at a 650 nm wavelength and 27.4 mW power. The highest variation of temperature in the seeds was obtained for the black-dyed condition, these variations being 5.56 and 9.28°C for crystalline and floury seeds, respec tively. Among the seeds, in the dyed condition, the floury seed had the lower non-radiative relaxation time, the higher optical absorption coefficient and a lower optical penetration length at the laser wavelength (650 nm). K e y w o r d s: maize, temperature, thermal camera, laser

Plants are the principal source of dietary iron (Fe) for most of Earth's population and Fe deficiency can lead to major health problems. Developing strategies to improve plant Fe content is a challenge because Fe is essential and toxic and therefore regulating Fe uptake is crucial for plant survival. Acquiring soil Fe relies on complex regulatory events that occur in root epidermal cells. We review recent advances in elucidating many aspects of the regulation of Fe acquisition. These include the expanding protein network involved in FER-LIKE IRON DEFICIENCY INDUCED TRANSCRIPTION FACTOR (FIT)-dependent gene regulation and novel findings on the intracellular trafficking of the Fe transporter IRON-REGULATED TRANSPORTER 1 (IRT1). We outline future challenges and propose strategies, such as exploiting natural variation, to further expand our knowledge.

In this paper, we present an alternative technique for the demodulation of single interferograms using a sliding 2-D continuous wavelet transform (2-D CWT) method. The sliding strategy proposed in this work is used with two purposes: to reduce the processing time when the 2-D CWT is applied, and to solve the problem of the phase ambiguity when closed fringes are present. Experimental results with real and simulated interferograms show that the proposed multiresolution method is a proper alternative for many applications of interferogram demodulation with closed fringes.

In this paper a measurement methodology is proposed to avoid some limitations of conventional front photopyroelectric technique for thermal effusivity measurements, related to the necessity of calibration using a reference sample. The same pyroelectric sensor heated at high frequencies acts as a reference sample. The proposed method is tested with measurements of thermal effusivity in liquid samples.

Endomembrane protein trafficking has emerged as important means of regulating stress responses in plants. The Arabidopsis SNX1 protein is involved in recycling the iron transporter IRT1, thus promoting its presence at the plasma membrane. SNX1 and its interacting partners undergo stress-related regulation at both transcriptional and posttranslational level, which may include differential regulation at tissue level. Based on this, we explore the tissue-specific regulation of iron import, specifically concentrating on the factors involved in the expression and recycling of IRT1 in root tissues. We propose that different processes affecting IRT1 regulation may lead to similar outcomes, allowing for fine-tuning iron acquisition and distribution.

Summary Nutrient acquisition is entangled with growth and stress in sessile organisms. The bHLH transcription factor FIT is a key regulator of Arabidopsis iron (Fe) acquisition and post-translationally activated upon low Fe. We identified CBL-INTERACTING PROTEIN KINASE CIPK11 as a FIT interactor. Cytosolic Ca2+ concentration and CIPK11 expression are induced by Fe deficiency. cipk11 mutant plants display compromised root Fe mobilization and seed Fe content. Fe uptake is dependent on CBL1/CBL9. CIPK11 phosphorylates FIT at Ser272, and mutation of this target site modulates FIT nuclear accumulation, homo-dimerization, interaction with bHLH039, and transcriptional activity and affects the plant’s Fe-uptake ability. We propose that Ca2+-triggered CBL1/9-mediated activation of CIPK11 and subsequent phosphorylation of FIT shifts inactive into active FIT, allowing regulatory protein interactions in the nucleus. This biochemical link between Fe deficiency and the cellular Ca2+ decoding machinery represents an environment-sensing mechanism to adjust nutrient uptake.

The sessile lifestyle of plants requires them to cope with stresses in situ. Plants overcome abiotic stresses by altering structure/morphology, and in some extreme conditions, by compressing the life cycle to survive the stresses in the form of seeds. Genetic and molecular studies have uncovered complex regulatory processes that coordinate stress adaptation and tolerance in plants, which are integrated at various levels. Investigating natural variation in stress responses has provided important insights into the evolutionary processes that shape the integrated regulation of adaptation and tolerance. This review primarily focuses on the current understanding of how transcriptional, post-transcriptional, post-translational, and epigenetic processes along with genetic variation orchestrate stress responses in plants. We also discuss the current and future development of computational tools to identify biologically meaningful factors from high dimensional, genome-scale data and construct the signaling networks consisting of these components. Published version false (Extension publication?)

Protein sorting in the endomembrane system is responsible for the coordination of cellular functions. Plant intracellular trafficking has its own unique features, which include specific regulatory aspects of endosomal sorting and recycling of cargo proteins, mediated by the retromer complex. Recent work has led to significant progress in understanding the role of Arabidopsis retromer subunits in recycling vacuolar sorting receptors and plasma membrane proteins. As a consequence, members of the sorting nexin (SNX) protein family and their interaction partners have emerged as critical protein trafficking regulators, in particular with regard to adaptation to environmental change, such as temperature fluctuations and nutrient deficiency. In this Review, we discuss the known and proposed functions of the comparatively small Arabidopsis SNX protein family. We review the available information on the role of the three Bin-Amphiphysin-Rvs (BAR)-domain-containing Arabidopsis thaliana (At)SNX proteins and discuss their function in the context of their potential participation in the plant retromer complex. We also summarize the role of AtSNX1-interacting proteins in different aspects of SNX-dependent protein trafficking and comment on the potential function of three novel, as yet unexplored, Arabidopsis SNX proteins.

Our knowledge of vacuole biogenesis and the transport of proteins to the vacuole has advanced consistently over the last 30 years. In meristematic cells, the tonoplast appears to develop directly out of the endoplasmic reticulum ([Viotti et al., 2013][1]). Once it is established as a functioning

A smart illumination system for greenhouses and growing rooms capable of controlling the quantity and quality of light emitted by a number of LEDs is described. The system uses lamps containing blue and red LEDs programmed to emit various spectra at sixteen different frequencies and ten different pulse widths. The performance of the system is evaluated by determining the effect of pulsed light emission at different frequencies with a pulse width of 50% on tomato plants ( Lycopersicon esculentum). The results show that low frequencies (0.1, 1, 10 Hz) have higher quantum efficiency in photosystem II compared to higher frequencies (50 and 100 kHz) and continuous light. They also show that the electron transport rate decreases when the frequency of pulses increases.

A smart illumination system (ILSys) to study the growth process of green plants was developed. This system can control commercial LED lamps that can be turned on to emit continuous or pulsed light. ILSys was applied to determine the effect of pulsed light on the fluorescence emission in tomato plants (Lycopersicon esculentum). The fluorescence emission was measured when the plants were radiated with continuous light and light pulsed at different frequencies (0.1Hz, 1Hz, 10Hz, 50Hz, 100Hz, 500Hz, 1kHz, 5kHz, 10kHz, 50kHz and 100kHz). For the experiment three red and blue-LEDs lamps and a Pulse Modulated Chlorophyll Fluorescence Monitoring System were connected to the ILSys. Results showed the highest quantum efficiencies in the Photosystem II (@fPSII) was obtained when applying pulse frequency of 0.1Hz and 1Hz with 50% light pulse width. ILSys allowed the finding of a significant difference in the response of the quantum efficiency in the Photosystem II between pulsed light at a number of frequencies and continuous light.

The effects of pulsed light based-LEDs at eleven frequencies (0.1, 1, 10, 50, 100, 500 Hz, 1, 5, 10, 50 and 100 kHz) programmed at 50 % duty cycle were analyzed, obtaining important parameters of the fluorescence emission of chlorophyll such as: maximum fluorescence (Fm′), minimum fluorescence, the fluorescence emission in steady state, maximum efficiency of PSII (Fv′/Fm′), the fraction of PSII centers that are open, photochemical quenching, nonphotochemical quenching (NPQ), quantum efficiency of photosystem II (ΦPSII), electron transport rate (ETR) and quantum yield of CO2 assimilation (ϕCO2). For the study and validation of the results obtained in the experiments, the analysis of variance (ANOVA) was applied 0for each parameter with confidence intervals of 95 %. The results show that the frequencies of pulsed light had positive and negative effects on the fluorescence parameters with respect to the control treatment (continuous light). The frequencies that generated the best performance of Fv′/Fm′, NPQ, ΦPSII, ETR, ϕCO2 in tomato plants were 0.1, 1, 100 Hz, and 1 kHz. The increase obtained in these parameters can represent an optimal growth and productivity conditions for optimal energy consumption.

The so-called electropyroelectric technique uses amplitude modulated electrical current as excitation source of thermal waves in a pyroelectric sensor in contact with a liquid sample. Here it is showed that this technique allows absolute measurements of the thermal effusivity using the quotient of the signal amplitudes obtained from the inverse and the direct configurations. In this way the influence on the experimental results of the frequency dependent instrumental transfer function can be straightforwardly eliminated. Measurements performed in various test liquid samples are presented that demonstrate the good accuracy and usefulness of the method.

Signaling mediated by reactive oxygen species (ROS) has emerged as a key component of plants' responses to environmental stress. The ROS-regulated transcription factor ZAT12 was revealed as a negative regulator of iron (Fe) deficiency responses through its direct interaction with the bHLH protein FIT. In the epidermis of the early root differentiation zone, ZAT12 stability depended on the presence of the ZAT12 EAR motif. It was concluded that ZAT12 may be the target of 2 alternative degradation pathways. Here, we present a model aiming to explain the regulatory mechanisms by which ZAT12 could be targeted for degradation and to predict the types of potential regulators involved. In addition to an E3 ubiquitin ligase, we predict 2 critical regulatory factors, namely a protein interacting with the ZAT12 EAR motif and a ROS-responsive regulatory protein.

This article deals with the photopyroelectric (PPE) technique, a useful tool for optical spectroscopy and for measurement of thermal properties of condensed mater samples, in particular in the liquid state. It has been demonstrated here that, for constant laser power, the pyroelectric (PE) sensor response does not depend on the spatial distribution of the laser beam intensity. As a consequence of this, it follows that in the case of measurements in voltage mode, the signal amplitude is inversely proportional to the active area of the sensor. Then, if the active area is diminished, leaving the total area of the sensor constant, it is possible to increase the pyroelectric signal. On the basis of this result, a method is proposed to enhance the PPE signal measured in voltage mode by means of the optimization in the configuration of the sensor metal electrodes. It was experimentally demonstrated that using this improved technique, it is possible to increase up to ten times the amplitude of the signal without electrical noise rise.

Recently the so-called electropyroelectric technique for thermal characterization of liquids has been proposed (Ivanov et al., J. Phys. D: Appl. Phys. 43, 225501 (2010)). In this method a pyroelectric sensor, in good thermal contact with the investigated sample, is heated by passing an amplitude-modulated electrical current through the electrical contacts. As a result of the heat dissipated to the sample, the pyroelectric signal measured as a voltage drop across the electrical contacts changes in a periodical way. The amplitude and phase of this signal can be measured by lock-in detection as a function of the electrical current modulation frequency. Because the signal amplitude and phase depend on the thermal properties of the sample, these can be determined straightforwardly by fitting the experimental data to a theoretical model based on the solution of the heat diffusion equation with proper boundary conditions. In general, the experimental conditions are selected so that the thermal effusivity becomes the measured magnitude. The technique has the following handicap. As the result of heating and wear of the metal coating layers (previously etched to achieve a serpentine form) with time, their electrical resistance changes with time, so that the heat power dissipated by the Joule effect can vary, and thermal effusivity measurement can become inaccurate. To avoid this problem in this study, a method is proposed that allows maintaining stable the Joule dissipated power. An electronic circuit is designed whose stability and characteristics are investigated and discussed.

The implementation of an automated system based on the hot-wire technique is described for the measurement of the thermal conductivity of liquids using equipment easily available in modern physics laboratories at high schools and universities (basically a precision current source and a voltage meter, a data acquisition card, a personal computer and a high purity platinum wire). The wire, which is immersed in the investigated sample, is heated by passing a constant electrical current through it, and its temperature evolution, ΔT, is measured as a function of time, t, for several values of the current. A straightforward methodology is then used for data processing in order to obtain the liquid thermal conductivity. The start point is the well known linear relationship between ΔT and ln(t) predicted for long heating times by a model based on a solution of the heat conduction equation for an infinite lineal heat source embedded in an infinite medium into which heat is conducted without convective and radiative heat losses. A criterion is used to verify that the selected linear region is the one that matches the conditions imposed by the theoretical model. As a consequence the method involves least-squares fits in linear, semi-logarithmic (semi-log) and log–log graphs, so that it becomes attractive not only to teach about heat transfer and thermal properties measurement techniques, but also as a good exercise for students of undergraduate courses of physics and engineering learning about these kinds of mathematical functional relationships between variables. The functionality of the experiment was demonstrated by measuring the thermal conductivity in samples of liquids with well known thermal properties.

During the late stages of seed development, the embryo patterning program is completed and maturation is initiated. One of the main events during the maturation phase is the acquisition of dormancy, characterized by the failure of a normally developed seed to germinate precociously. Dormancy is controlled by a complex regulatory mechanism that involves the phytohormone gibberellin (GA) and the transcription factor FUSCA3 (FUS3). Here, we demonstrate the importance of the previously characterized GA regulator EFFECTOR OF TRANSCRIPTION2 (AtET2) for correct seed development. We show that entering the maturation phase, seeds of the et2-1 mutant, which contain a non-functional AtET2 gene, fail to induce dormancy. This correlates well with the observed activity pattern of the AtET2 promoter, which is active in the maturing embryo. AtET2 action during seed development is dependent on a complex interaction with GA and the FUS3 gene, the latter evidenced by the phenotypes of the et2-1 fus3-T double mutant. We show that in vitro expressed AtET2 protein can bind to both linear and supercoiled DNA without any obvious sequence preference. This suggests that, within a larger protein complex, AtET2 might be required for the correct positioning upon the DNA.

In this work we present a phase to (x,y,z)-coordinates transformation method for the calibration of a fringe projection profilometer. Our technique is divided in two parts: (1) phase to z transformation (for axial calibration) based on the typical polynomial fitting which uses a flat plane placed at several z positions to measure the phase of the projected fringes. (2) Phase to x and y transformation (for transverse calibration) based on the use of a crossed gratings pattern and a Fourier phase measurement method to determine x and y coordinates at several z positions. As will be shown the use of the crossed gratings pattern and the Fourier phase measurement method for transverse calibration is advantageous in several aspects: an unique image can give us x and y information at once. It provides x and y coordinates at each pixel in the image avoiding the use of interpolation methods. We present some experimental results and explain the viability of the proposed technique.

Iron is an essential element for life on Earth and its shortage, or excess, in the living organism may lead to severe health disorders. Plants serve as the primary source of dietary iron and improving crop iron content is an important step towards a better public health. Our review focuses on the control of iron acquisition in dicotyledonous plants and monocots that apply a reduction-based strategy in order to mobilize and import iron from the rhizosphere. Achieving a balance between shortage and excess of iron requires a tight regulation of the activity of the iron uptake system. A number of studies, ranging from single gene characterization to systems biology analyses, have led to the rapid expansion of our knowledge on iron uptake in recent years. Here, we summarize the novel insights into the regulation of iron acquisition and internal mobilization from intracellular stores. We present a detailed view of the main known regulatory networks defined by the Arabidopsis regulators FIT and POPEYE (PYE). Additionally, we analyze the root and leaf iron-responsive regulatory networks, revealing novel potential gene interactions and reliable iron-deficiency marker genes. We discuss perspectives and open questions with regard to iron sensing and post-translational regulation.

A principal objective in life sciences is the visualization of biochemical processes. Fluorescence-based techniques are widely used to demonstrate transport of relevant substances across cellular membranes. In this paper we report a novel noninvasive, real-time fluorescence lifetime imaging microscopy method for visualizing uptake and release of divalent copper ions (Cu(2+) ) in vivo. For this purpose, we employed a green fluorescent protein (GFP) form able to change its fluorescence lifetime upon Cu(2+) binding. We demonstrate that this technique is selective for Cu(2+) . We show the reversible decrease of the fluorescence lifetime of GFP from 2.2 to 1.6 ns in Escherichia coli and from 1.8 to 1.3 ns in root cells of Arabidopsis after the addition of Cu(2+) . Cu(2+) uptake of epidermal tobacco cells leads to a drop of the GFP lifetime from 2.5 to 2.2 ns. In summary, the spatially resolved visualization of Cu(2+) distribution in vivo is demonstrated in prokaryote and eukaryote cells.

A pyroelectric sensor can be seen as a layered system consisted on a pyroelectric material sandwiched between two thin metal layers acting as electrical contacts for measurements of the voltage drop that can be induced by heating. This kind of sensor can be used as a detector of electromagnetic radiation but also for thermal characterization of materials using the photopyroelectric technique. In this work we perform a theoretical analysis based in the so-called thermal wave approach to show that, when this sensor is heated periodically by the absorption of intensity modulated light by one of the metalized surfaces, while the other metal surface is in contact with a liquid sample, the resulting pyroelectric voltage signal amplitude enhances respecting the one resulting from the bare sensor, for certain values of the modulation frequency. This contradicts the intuitively expectation based in the assumption that the sample provides a new channel for heat conduction, thereby decreasing the pyroelectric temperature. We will show that the back and forth propagation and the superposition of thermal waves through the metal coatings must be taken into account in order to explain the observed behavior. The proposed model was experimentally tested for water and glycerin samples, and using a polyvinylidene difluoride (PVDF) polymer film, with Ni–Cu metal electrodes, as a pyroelectric sensor.

The understanding of cellular processes and functions and the elucidation of their physiological mechanisms is an important aim in the life sciences. One important aspect is the uptake and the release of essential substances as well as their interactions with the cellular environment. As green fluorescent protein (GFP) can be genetically encoded in cells it can be used as an internal sensor giving a deeper insight into biochemical pathways. Here we report that the presence of copper(II) ions leads to a decrease of the fluorescence lifetime (τ fl) of GFP and provide evidence for Forster resonance energy transfer (FRET) as the responsible quenching mechanism. We identify the His6-tag as the responsible binding site for Cu2 + with a dissociation constant K d = 9 ±2 μM and a Forster radius R 0 = 2.1 ±0.1 nm. The extent of the lifetime quenching depends on [Cu2 + ] which is comprehended by a mathematical titration model. We envision that Cu2 + can be quantified noninvasively and in real-time by measuring τ fl of GFP.

In this paper the use of the photothermal radiometry technique in the frequency domain (PRTF) and the use of multivariate methods in the study of two types of skin phantoms: (a) one in which skin pigmentation was simulated dyeing the gel phantom and (b) the other consists of exposure of animal skin samples to different degrees of thermal damage. In experiment (a), gel phantoms were prepared with different concentrations of methylene blue (MB). The mean values of the radiometry signal (RS) show significant differences in only those cases in which changes in the concentration of MB were higher than 0.38 mM. This result was confirmed with a t test for independent samples of the data (p < 0.05). The mean values of the amplitude and phase signal do not permit discrimination between phantoms with changes in pigmentation equal to or lower than this value. However, principal component analysis (PCA) demonstrated that it is possible to discriminate between phantoms with changes in molar concentration equal to 0.38 mM (for the phase signal). In the case of experiment (b), the following four groups of pork skin samples were analyzed: one consists of samples of fresh skin, while the other three consist of samples exposed to thermal damage at 45 °C (the exposure time was 4 s) and 80 °C (exposure times were 4 s and 8 s, respectively). The mean values of the RS for each group of samples did not show a clear visual discrimination. However, the t test for independent samples applied to the data demonstrated significant differences only between fresh skin and skin exposure to thermal damage at 80 °C (with exposure times of 4 s and 8 s). PCA was used to discriminate between the four different skin samples.

The aim of the present study was to obtain qualitative content of carotenoids in the leaves of maize seedlings the seeds of which were irradiated by the 650 nm diode laser. Four different irradiation doses have been applied; and the control (non-irradiated) seeds used for comparison purposes. Optical absorption spectra of leaves from seedlings were recorded by means of photoacoustic spectroscopy in the 400 to 500 nm range. The experimental unit comprised 10 seeds, among which one seedling was selected at random for each repetition. Data was subjected to variance analysis (ANOVA) using the SAS GLM procedures (SAS, 1998 version). The ANOVA revealed significant (p < 0.05) differences among treatments in the 471 to 478 nm range.

This paper describes an alternative methodology to determine the thermal effusivity of a liquid sample using the recently proposed electropyroelectric technique, without fitting the experimental data with a theoretical model and without having to know the pyroelectric sensor related parameters, as in most previous reported approaches. The method is not absolute, because a reference liquid with known thermal properties is needed. Experiments have been performed that demonstrate the high reliability and accuracy of the method with measurement uncertainties smaller than 3%.

Single Cell Gel Electrophoresis (SCGE) or Comet assay is a very sensitive method for assessing damages in DNA on a single cell level. It has found many applications in fields where genotoxic activity could be an issue. In environmental monitoring, health care, food industry Comet assay is used with increasing popularity. For verifying the results obtained by this method many parameters could be monitored. To that end several software packages exist. In the conditions that we are suggesting one more parameter could be measured - comet shape. We argue that this parameter could be an advantage of the Comet Assay when the way of DNA damaging needs to be predicted.

Розглядаються два методи блокування шкідливих “капканів”, які реалізовані за допомогою лічильників та на основі часу з початку запуску програми.

Total of 35 shells were found and identified during the archeological excavations 2012-2014, studied Slavic dwellings-dugouts which reuse earlier structures of the roman forum inPlovdiv, Bulgaria. From both localities shells from 7 species of snails were found. Most of the shellswere from two species: Zebrina detrita and Helix philibinensis – total of 28 shells, other wererepresented by 1-3 specimens per species. The lack of shell materials from Eobania vermiculatasupports the theory of later introduction of this species in the city of Plovdiv.

We have determined and compared nucleotide excision repair capability of several rat tissues by a method based on restoration of the transformation activity of UV-irradiated pBlueScript by incubation in repair-competent protein extracts. After 3 h of incubation, plasmid DNA was isolated and used to transform competent Escherichia coli cells. Damaged plasmids showed low transformation efficiency prior to incubation in repair-competent extracts. After incubation the transformation efficiency was restored to different extents permitting calculation of the repair capacity of the extracts. Our results showed that rapidly proliferating tissues such as liver, kidney and testis showed higher nucleotide excision repair capacity than slowly proliferating tissues such as heart, muscle, lung and spleen. When liver and splenocytes were stimulated to proliferation by partial hepatectomy and mitogen stimulation, their repair capability increased in parallel with the respective proliferative rates.

Copper ions play a fundamental role in plant metabolism where its uptake and distribution within the organism is highly regulated, allowing the cells to sustain an adequate concentration. Shortage or excess of Cu can cause severe damage to the organisms endangering their survival. We recently reported a non-invasive method to follow the intracellular uptake of bivalent copper ion concentration by fluorescence lifetime microscopy of green fluorescent protein within plant cells. Measuring the fluorescence lifetime has the advantage of being independent on the fluorophore concentration and the excitation intensity. The use of GFP is beneficial because the protein can be introduced nondestructively. Here, we discuss the benefits of this approach as well as the possibility of applying this concept for the investigation of Cu redistribution and storage at the subcellular level. The fluorescence lifetime-encoded microscopic images are envisioned to map the copper distribution within plant cells not only qualitatively but even quantitatively. Time-lapse microscopy enables the following of cellular processes and the study of relevant transport mechanisms of copper in plant cells. Perspectives and necessary improvements are discussed.

Dicotyledonous plants growing under limited iron availability initiate a response resulting in the solubilization, reduction, and uptake of soil iron. The protein factors responsible for these steps are transmembrane proteins, suggesting that the intracellular trafficking machinery may be involved in iron acquisition. In search for components involved in the regulation of Arabidopsis thaliana iron deficiency responses, we identified the members of the SORTING NEXIN (SNX) protein family. SNX loss-of-function plants display enhanced susceptibility to iron deficiency in comparison to the wild type. The absence of SNX led to reduced iron import efficiency into the root. SNX1 showed partial colocalization with the principal root iron importer IRON-REGULATED TRANSPORTER1 (IRT1). In SNX loss-of-function plants, IRT1 protein levels were decreased compared with the wild type due to enhanced IRT1 degradation. This resulted in diminished amounts of the IRT1 protein at the plasma membrane. snx mutants exhibited enhanced iron deficiency responses compared with the wild type, presumably due to the lower iron uptake through IRT1. Our results reveal a role of SNX1 for the correct trafficking of IRT1 and, thus, for modulating the activity of the iron uptake machinery.

Self-incompatibility (SI) has emerged as an evolutionary strategy to enhance the genetic variability of plant species. In Brassica, it is controlled by a single multiallelic locus, the S-locus, encoding a receptor kinase (SRK) expressed in the stigma papilla cells and its ligand, a small protein (SCR) located in the pollen coat. Pollen rejection is achieved only when the receptor recognizes SCR coming from the same S-allele. If a single papilla cell is simultaneously pollinated by a self- and a cross-pollen grain, it is capable of distinguishing between the two and responding accordingly, rejecting self while accepting cross pollen. This phenomenon reveals that SI response is strictly localized and does not involve the whole papilla cell. It also suggests that the distribution of SRK inside the cell may play an important role in regulating this dual response. We have recently demonstrated that SRK is mostly intracellular, only small amounts being present in distinct domains of the plasma membrane (PM), where interaction with SCR occurs. Following ligand recognition, the receptor-ligand complex is endocytosed and degraded. Based on this, we propose a model of the significance of SRK intracellular trafficking for the functioning and specificity of SI response.

This paper shows method for realization of hidden “trap” in the beginning of malicious code with using of time of program., Предлагается метод реализации скрытого “капкана” для старта вредительского кода с использованием содержания системных часов., Пропонується метод реалізації прихованого “капкана” для старту шкідливого коду з використанням змісту системних годинників.

Reliable and accurate testing methods are essential to guiding the polishing process during the figuring of optical telescope mirrors. With the natural advancement of technology, the procedures and instruments used to carry out this delicate task have consistently increased in sensitivity, but also in complexity and cost. Fortunately, throughout history, the Foucault knife-edge test has shown the potential to measure transverse aberrations in the order of the wavelength, mainly when described in terms of physical theory, which allows a quantitative interpretation of its characteristic shadowmaps. Our previous publication on this topic derived a closed mathematical formulation that directly relates the knife-edge position with the observed irradiance pattern. The present work addresses the quite unexplored problem of the wavefront's gradient estimation from experimental captures of the test, which is achieved by means of an optimization algorithm featuring a proposed ad hoc cost function. The partial derivatives thereby calculated are then integrated by means of a Fourier-based algorithm to retrieve the mirror's actual surface profile. To date and to the best of our knowledge, this is the very first time that a complete mathematical-grounded treatment of this optical phenomenon is presented, complemented by an image-processing algorithm which allows a quantitative calculation of the corresponding slope at any given point of the mirror's surface, so that it becomes possible to accurately estimate the aberrations present in the analyzed concave device just through its associated foucaultgrams.

From the Bayesian regularization theory we derive a quadratic cost-function for integrating wave-front gradient fields. In the proposed cost-function, the term of conditional distribution uses a central-differences model to make the estimated function well consistent with the observed gradient field. As will be shown, the results obtained with the central-differences model are superior to the results obtained with the backward-differences model, commonly used in other integration techniques. As a regularization term we use an isotropic first-order differences Markov Random-Field model, which acts as a low-pass filter reducing the errors caused by the noise. We present simulated and real experiments of the proposal applied in the Foucault test, obtaining good results.

In a recent paper published in this journal [R. Ivanov et al., Rev. Sci. Instrum. 86, 064902 (2015)], a methodology free of fitting procedures for determining the thermal effusivity of liquids using the electropyroelectric technique was reported. Here the same measurement principle is extended to the well-known photopyroelectric technique. The theoretical basis and experimental basis of the method are presented and its usefulness is demonstrated with measurements on test samples.

Self-incompatibility (SI) is an archetypal cell-to-cell communication system in which self-pollen is rejected to prevent inbreeding. In crucifers (or Brassicaceae family), the pollen SI determinant is a small peptide, the S-LOCUS CYSTEINE-RICH PROTEIN (SCR, also known as SP11). During self-pollination, SCR binds to the extracellular domain of its cognate stigmatic receptor, the S-LOCUS RECEPTOR KINASE (SRK). This initiates a signaling cascade leading to self-pollen inhibition. The genes encoding both SI determinants are tightly linked in a multiallelic genomic region defined as the S-locus. Among S alleles, SCR shows extreme variability, and few residues in the protein have been reported to be critical for its specificity. In a heterozygous situation, SCR alleles display complex dominance relationships based on the silencing of certain SCR genes. This chapter provides a summary of the role of SCR in SI, the relationships between SCR alleles, and the role SCR loss played in the model plant Arabidopsis thaliana.

Separation of functions within the cytoplasm of the eukaryotic cell has resulted in the development of a highly dynamic network of membranous compartments. Among them, the endosomes represent a crossing point for the major cellular trafficking pathways. Indeed, they are responsible for the communication between the cellular compartments, as well as between the cell and its environment, through the regulation of signalling networks and transport of material. The proteins from the sorting nexin (SNX) family have an established role as major regulators of endosomal protein transport in mammals, insects, worms, and yeast. By contrast, research on plant SNXs has been initiated relatively recently. Despite that, the accumulated knowledge suggests that plant SNXs may have evolved different mechanisms of action and might work in a different protein complex environment, compared to their yeast and animal counterparts. In this chapter, we highlight both common and specific characteristics of plant SNX protein function and regulation in comparison to the well-studied mammalian SNX machinery.

A principal objective in life sciences is the visualization of biochemical processes. Fluorescence-based techniques are widely used to demonstrate transport of relevant substances across cellular membranes. In this paper we report a novel noninvasive, real-time fluorescence lifetime imaging microscopy method for visualizing uptake and release of divalent copper ions (Cu(2+) ) in vivo. For this purpose, we employed a green fluorescent protein (GFP) form able to change its fluorescence lifetime upon Cu(2+) binding. We demonstrate that this technique is selective for Cu(2+) . We show the reversible decrease of the fluorescence lifetime of GFP from 2.2 to 1.6 ns in Escherichia coli and from 1.8 to 1.3 ns in root cells of Arabidopsis after the addition of Cu(2+) . Cu(2+) uptake of epidermal tobacco cells leads to a drop of the GFP lifetime from 2.5 to 2.2 ns. In summary, the spatially resolved visualization of Cu(2+) distribution in vivo is demonstrated in prokaryote and eukaryote cells.

International audience; More than half of the flowering plants have a sophisticated mechanism for self-pollen rejection, named self-incompatibility (SI). In Brassicaceae, recognition specificity is achieved by the interaction of the stigmatic S-RECEPTOR KINASE (SRK) and its ligand S-LOCUS CYSTEINE-RICH PROTEIN (SCR). Recent years have seen significant advances in understanding the SI response. Progress has been made on elucidating the regulation and function of proteins that act as either molecular partners of SRK or modulators of SI. Thus, modules controlling the specificity of the central receptor-ligand interaction have been identified on both SRK and SCR proteins. A role for intracellular protein trafficking in SI has also been demonstrated. Here, we integrate the novel findings into the existing model to present the current understanding of SI signaling.

Self-incompatibility (SI) has emerged as an evolutionary strategy to enhance the genetic variability of plant species. In Brassica, it is controlled by a single multiallelic locus, the S-locus, encoding a receptor kinase (SRK) expressed in the stigma papilla cells and its ligand, a small protein (SCR) located in the pollen coat. Pollen rejection is achieved only when the receptor recognizes SCR coming from the same S-allele. If a single papilla cell is simultaneously pollinated by a self- and a cross-pollen grain, it is capable of distinguishing between the two and responding accordingly, rejecting self while accepting cross pollen. This phenomenon reveals that SI response is strictly localized and does not involve the whole papilla cell. It also suggests that the distribution of SRK inside the cell may play an important role in regulating this dual response. We have recently demonstrated that SRK is mostly intracellular, only small amounts being present in distinct domains of the plasma membrane (PM), where interaction with SCR occurs. Following ligand recognition, the receptor-ligand complex is endocytosed and degraded. Based on this, we propose a model of the significance of SRK intracellular trafficking for the functioning and specificity of SI response.

A brief review of the state of the art in quantitative photoacoustic spectroscopy for thermally thick samples is presented. We demonstrate that it is possible to find an analytical expression that allows studying the optical absorption coefficient by combining the amplitude and phase of the photoacoustic signal, obtained from the heat diffusion model, for thermally thick samples.

The plant hormone gibberellin (GA) is known to modulate various aspects of plant cell differentiation and development. The current model of GA-mediated regulation is based on a de-repressible system and includes specific protein modification and degradation. HRT, a zinc finger protein from barley has been shown to have GA-dependent transcriptional repressing activity on the seed-specific alpha-amylase promoter [Raventos, D., Skriver, K., Schlein, M., Karnahl, K., Rogers, S.W., Rogers, J.C. and Mundy, J. 1998. J. Biol. Chem. 273: 23313-23320]. Here we report the characterization of a dicot homologue from Brassica napus (BnET) and provide evidence for its role in GA response modulation suggesting that this could be a conserved feature of this gene family. When BnET is ectopically expressed in either Arabidopsis or tobacco the phenotypes include dwarfism due to shorter internodes and late flowering, reduced germination rate, increased anthocyanin content and reduced xylem lignification as a marker for terminal cell differentiation. Transient expression in protoplasts supports the notion that this most likely is due to a transcriptional repression of GA controlled genes. Finally, histological analysis showed that in contrast to other GA deficient mutants the shorter internodes were due to fewer but not smaller cells, suggesting a function of BnET in GA-mediated cell division control.

Preliminary studies of erythrocyte sedimentation rate in real time, using the inverse photopyroelectric method, are presented. The advantages of this methodology is that the sample is not illuminated directly and that the process can be monitored through the measurement of the thermal contact between a pyroelectric sensor and the blood. The thermal effusivity was determined using a one‐dimensional heat diffusion model, where the pyroelectric sensor is thermally thin and the blood is thermally thick. It was demonstrated that during the process, the thermal effusivity decreases in the region of the contact between the blood and the pyroelectric sensor. The usefulness of these results in real time monitoring using photothermal techniques is discussed.