Upstream Sensors Research Articles

807: CD36 expression in trophoblasts: implications for preeclampsia

out using SP600125 (different concentrations: 10, 20, or 50 uM) prior to above stress stimulations. Expressional levels of these stress response elements and MAPK signal pathway effectors were assessed via western blots. RESULTS: Gadd45a protein was induced together with the downstream phospho-JNK activation in response to different preeclampsia associated stressors including hypoxia, inflammatory cytokines IL-6, TNFand Angiotensin II. The activation of phosphor-JNK was inhibited by SP600125 at 10uM, 10uM, 50uM and 10uM for hypoxia, IL-6, TNFand Angiotensin II respectively. CONCLUSION: While evaluating pathways to preeclampsia, our data using placental explants model provides evidence that preeclampsia associated stressors that reported to be increased in preeclampsia activate phospho-JNK together with its up-stream stress sensor protein Gadd45a. Further evidence, on these basic pathways ending clinically with preeclampsia will be obtained by in-vivo experiments using Gadd45a knockout mice.

Regulation of Manganese Antioxidants by Nutrient Sensing Pathways in Saccharomyces cerevisiae

In aerobic organisms, protection from oxidative damage involves the combined action of enzymatic and nonproteinaceous cellular factors that collectively remove harmful reactive oxygen species. One class of nonproteinaceous antioxidants includes small molecule complexes of manganese (Mn) that can scavenge superoxide anion radicals and provide a backup for superoxide dismutase enzymes. Such Mn antioxidants have been identified in diverse organisms; however, nothing regarding their physiology in the context of cellular adaptation to stress was known. Using a molecular genetic approach in Bakers' yeast, Saccharomyces cerevisiae, we report that the Mn antioxidants can fall under control of the same pathways used for nutrient sensing and stress responses. Specifically, a serine/threonine PAS-kinase, Rim15p, that is known to integrate phosphate, nitrogen, and carbon sensing, can also control Mn antioxidant activity in yeast. Rim15p is negatively regulated by the phosphate-sensing kinase complex Pho80p/Pho85p and by the nitrogen-sensing Akt/S6 kinase homolog, Sch9p. We observed that loss of either of these upstream kinase sensors dramatically inhibited the potency of Mn as an antioxidant. Downstream of Rim15p are transcription factors Gis1p and the redundant Msn2/Msn4p pair that typically respond to nutrient and stress signals. Both transcription factors were found to modulate the potency of the Mn antioxidant but in opposing fashions: loss of Gis1p was seen to enhance Mn antioxidant activity whereas loss of Msn2/4p greatly suppressed it. Our observed roles for nutrient and stress response kinases and transcription factors in regulating the Mn antioxidant underscore its physiological importance in aerobic fitness.

Multiple TORC1-Associated Proteins Regulate Nitrogen Starvation-Dependent Cellular Differentiation in Saccharomyces cerevisiae

BackgroundThe budding yeast Saccharomyces cerevisiae undergoes differentiation into filamentous-like forms and invades the growth medium as a foraging response to nutrient and environmental stresses. These developmental responses are under the downstream control of effectors regulated by the cAMP/PKA and MAPK pathways. However, the upstream sensors and signals that induce filamentous growth through these signaling pathways are not fully understood. Herein, through a biochemical purification of the yeast TORC1 (Target of Rapamycin Complex 1), we identify several proteins implicated in yeast filamentous growth that directly associate with the TORC1 and investigate their roles in nitrogen starvation-dependent or independent differentiation in yeast.MethodologyWe isolated the endogenous TORC1 by purifying tagged, endogenous Kog1p, and identified associated proteins by mass spectrometry. We established invasive and pseudohyphal growth conditions in two S. cerevisiae genetic backgrounds (Σ1278b and CEN.PK). Using wild type and mutant strains from these genetic backgrounds, we investigated the roles of TORC1 and associated proteins in nitrogen starvation-dependent diploid pseudohyphal growth as well as nitrogen starvation-independent haploid invasive growth.ConclusionsWe show that several proteins identified as associated with the TORC1 are important for nitrogen starvation-dependent diploid pseudohyphal growth. In contrast, invasive growth due to other nutritional stresses was generally not affected in mutant strains of these TORC1-associated proteins. Our studies suggest a role for TORC1 in yeast differentiation upon nitrogen starvation. Our studies also suggest the CEN.PK strain background of S. cerevisiae may be particularly useful for investigations of nitrogen starvation-induced diploid pseudohyphal growth.

Damage to the glycoshield activates PMT‐directed O‐mannosylation via the Msb2–Cek1 pathway in Candida albicans

Protein-O-mannosyltransferases (Pmt) transfer mannosyl residues to secretory proteins. Five isoforms of Pmt proteins in the human fungal pathogen Candida albicans have distinct functions in growth, morphogenesis and antifungal resistance. We found that PMT genes encoding the major isoforms Pmt1, Pmt2, Pmt4 are regulated differently in response to impaired glycostructures. While the PMT1 transcript level increased in cell wall mutants and under inhibition of N-glycosylation by tunicamycin, PMT2 and PMT4 transcripts were upregulated only by inhibition of Pmt1 activity. Reporter fusions revealed specific promoter sequences to be required for PMT1 repression in undamaged cells, which was de-repressed by tunicamycin. Constitutive PMT1 de-repression was observed in mutants lacking the Cek1 MAP kinase and its upstream sensor Msb2. In contrast, in msb2 and cek1 mutants, upregulation of PMT2/PMT4 by Pmt1 inhibition did not occur and basal expression of both transcripts were decreased. We identified Ace2 as a novel transcription factor, which upregulates PMT basal expression and induction in response to glycostructure damage. Mutants lacking Msb2, Cek1 and Ace2 were supersensitive to glycosylation and cell wall inhibitors. We propose that a Msb2, Cek1 and Ace2 signalling pathway addresses PMT genes as downstream targets and that different modes of regulation have evolved for PMT1 and PMT2/PMT4 genes.

Multiple Plant Surface Signals are Sensed by Different Mechanisms in the Rice Blast Fungus for Appressorium Formation

Surface recognition and penetration are among the most critical plant infection processes in foliar pathogens. In Magnaporthe oryzae, the Pmk1 MAP kinase regulates appressorium formation and penetration. Its orthologs also are known to be required for various plant infection processes in other phytopathogenic fungi. Although a number of upstream components of this important pathway have been characterized, the upstream sensors for surface signals have not been well characterized. Pmk1 is orthologous to Kss1 in yeast that functions downstream from Msb2 and Sho1 for filamentous growth. Because of the conserved nature of the Pmk1 and Kss1 pathways and reduced expression of MoMSB2 in the pmk1 mutant, in this study we functionally characterized the MoMSB2 and MoSHO1 genes. Whereas the Momsb2 mutant was significantly reduced in appressorium formation and virulence, the Mosho1 mutant was only slightly reduced. The Mosho1 Momsb2 double mutant rarely formed appressoria on artificial hydrophobic surfaces, had a reduced Pmk1 phosphorylation level, and was nonresponsive to cutin monomers. However, it still formed appressoria and caused rare, restricted lesions on rice leaves. On artificial hydrophilic surfaces, leaf surface waxes and primary alcohols-but not paraffin waxes and alkanes- stimulated appressorium formation in the Mosho1 Momsb2 mutant, but more efficiently in the Momsb2 mutant. Furthermore, expression of a dominant active MST7 allele partially suppressed the defects of the Momsb2 mutant. These results indicate that, besides surface hydrophobicity and cutin monomers, primary alcohols, a major component of epicuticular leaf waxes in grasses, are recognized by M. oryzae as signals for appressorium formation. Our data also suggest that MoMsb2 and MoSho1 may have overlapping functions in recognizing various surface signals for Pmk1 activation and appressorium formation. While MoMsb2 is critical for sensing surface hydrophobicity and cutin monomers, MoSho1 may play a more important role in recognizing rice leaf waxes.

A lightweight, self‐adaptive lock gate designation scheme for data collection in long‐thin wireless sensor networks

ABSTRACTConstrained by the physical environments, the long‐thin topology has recently been promoted for many practical deployments of wireless sensor networks (WSNs). In general, a long‐thin topology is composed of a number of long branches of sensor nodes, where along a branch each sensor node has only one potential parent node toward the sink node. Although data aggregation may alleviate excessive packet contention, the maximum payload size of a packet and the dynamically changing traffic loads may severely affect the amount of sensor readings that may be collected along a long branch of sensor nodes. In addition, many practical applications of long‐thin WSNs demand the exact sensor readings at each location along the deployment areas for monitoring and analysis purposes, so sensor readings may not be aggregated when they are collected. This paper proposes a lightweight, self‐adaptive scheme that designates multiple collection nodes, termed lock gates, along a long‐thin network to collect sensor readings sent from their respective upstream sensor nodes. The self‐adaptive lock gate designation scheme balances between the responsiveness and the congestion of data collection while mitigating the funneling effect. The scheme also dynamically adapts the designation of lock gates to accommodate the time‐varying sensor reading generation rates of different sensor nodes. A testbed of 100 Jennic sensor nodes is developed to demonstrate the effectiveness of the proposed lock gate designation scheme. Copyright © 2011 John Wiley & Sons, Ltd.

Applications of wavelet transform for analysis of freeway traffic: Bottlenecks, transient traffic, and traffic oscillations

This paper demonstrates the capabilities of wavelet transform (WT) for analyzing important features related to bottleneck activations and traffic oscillations in congested traffic in a systematic manner. In particular, the analysis of loop detector data from a freeway shows that the use of wavelet-based energy can effectively identify the location of an active bottleneck, the arrival time of the resulting queue at each upstream sensor location, and the start and end of a transition during the onset of a queue. Vehicle trajectories were also analyzed using WT and our analysis shows that the wavelet-based energies of individual vehicles can effectively detect the origins of deceleration waves and shed light on possible triggers (e.g., lane-changing). The spatiotemporal propagations of oscillations identified by tracing wavelet-based energy peaks from vehicle to vehicle enable analysis of oscillation amplitude, duration and intensity.

Flux Footprints in the Convective Boundary Layer: Large-Eddy Simulation and Lagrangian Stochastic Modelling

We investigated the flux footprints of receptors at different heights in the convective boundary layer (CBL). The footprints were derived using a forward Lagrangian stochastic (LS) method coupled with the turbulent fields from a large-eddy simulation model. Crosswind-integrated flux footprints shown as a function of upstream distances and sensor heights in the CBL were derived and compared using two LS particle simulation methods: an instantaneous area release and a crosswind linear continuous release. We found that for almost all sensor heights in the CBL, a major positive flux footprint zone was located close to the sensor upstream, while a weak negative footprint zone was located further upstream, with the transition band in non-dimensional upwind distances −X between approximately 1.5 and 2.0. Two-dimensional (2D) flux footprints for a point sensor were also simulated. For a sensor height of 0.158 z i, where z i is the CBL depth, we found that a major positive flux footprint zone followed a weak negative zone in the upstream direction. Two even weaker positive zones were also present on either side of the footprint axis, where the latter was rotated slightly from the geostrophic wind direction. Using CBL scaling, the 2D footprint result was normalized to show the source areas and was applied to real parameters obtained using aircraft-based measurements. With a mean wind speed in the CBL of U = 5.1 m s−1, convective velocity of w * = 1.37 m s−1, CBL depth of z i = 1,000 m, and flight track height of 159 m above the surface, the total flux footprint contribution zone was estimated to range from about 0.1 to 4.5 km upstream, in the case where the wind was perpendicular to the flight track. When the wind was parallel to the flight track, the total footprint contribution zone covered approximately 0.5 km on one side and 0.8 km on the other side of the flight track.

Expanding the Substantial Interactome of NEMO Using Protein Microarrays

Signal transduction by the NF-kappaB pathway is a key regulator of a host of cellular responses to extracellular and intracellular messages. The NEMO adaptor protein lies at the top of this pathway and serves as a molecular conduit, connecting signals transmitted from upstream sensors to the downstream NF-kappaB transcription factor and subsequent gene activation. The position of NEMO within this pathway makes it an attractive target from which to search for new proteins that link NF-kappaB signaling to additional pathways and upstream effectors. In this work, we have used protein microarrays to identify novel NEMO interactors. A total of 112 protein interactors were identified, with the most statistically significant hit being the canonical NEMO interactor IKKbeta, with IKKalpha also being identified. Of the novel interactors, more than 30% were kinases, while at least 25% were involved in signal transduction. Binding of NEMO to several interactors, including CALB1, CDK2, SAG, SENP2 and SYT1, was confirmed using GST pulldown assays and coimmunoprecipitation, validating the initial screening approach. Overexpression of CALB1, CDK2 and SAG was found to stimulate transcriptional activation by NF-kappaB, while SYT1 overexpression repressed TNFalpha-dependent NF-kappaB transcriptional activation in human embryonic kidney cells. Corresponding with this finding, RNA silencing of CDK2, SAG and SENP2 reduced NF-kappaB transcriptional activation, supporting a positive role for these proteins in the NF-kappaB pathway. The identification of a host of new NEMO interactors opens up new research opportunities to improve understanding of this essential cell signaling pathway.

Investigation on the determination of flow direction using two parallel cylindrical hot film sensors

Measurement of instantaneous air flow velocity with high frequency can be carried out by using a hot wire anemometer (HWA). HWA works on the basis of heat transfer rate from hot wire to the fluid flow, therefore directional identification of the air flow using hot wire anemometer is a difficult task. By using two parallel cylindrical hot film sensors a probe was built. By considering the wake and heat effect of the upstream sensor on the downstream sensor, direction of the air flow can be identified. In this work, the wake and heat effect resulting from the upstream sensor to the velocity measurement, by the downstream sensor was studied. This measured velocity is dependent of the following factors namely; air velocity, upstream sensor overheat ratio, distance between the two sensors and turbulence intensity of the flow. As a result it was found that the manufactured probe with sensor distance of 1 mm apart is capable of measuring reverse flow measurements of up to 20 m/s for a moderate turbulent flow.

Gadd45 proteins as critical signal transducers linking NF-kappaB to MAPK cascades.

The growth arrest and DNA damage-inducible 45 (Gadd45) proteins are a group of critical signal transducers that are involved in regulations of many cellular functions. Accumulated data indicate that all three Gadd45 proteins (i.e., Gadd45alpha, Gadd45beta, and Gadd45gamma) play essential roles in connecting an upstream sensor module, the transcription Nuclear Factor-kappaB (NF-kappaB), to a transcriptional regulating module, mitogen-activated protein kinase (MAPK). This NF-kappaB-Gadd45(s)-MAPK pathway responds to various kinds of extracellular stimuli and regulates such cell activities as growth arrest, differentiation, cell survival, and apoptosis. Defects in this pathway can also be related to oncogenesis. In the first part of this review, the functions of Gadd45 proteins, and briefly NF-kappaB and MAPK, are summarized. In the second part, the mechanisms by which Gadd45 proteins are regulated by NF-kappaB, and how they affect MAPK activation, are reviewed.

CONDUCTANCE SENSOR FOR MEASUREMENT OF THE FLUID WATER CUT AND FLOW RATE IN PRODUCTION WELLS

A new type of conductance sensor has been studied to simultaneously measure both flow rate and water cut of oil-water two-phase fluid in production wells. The water cut is obtained by the ratio of the conductivity of the oil-water mixture fluid to that of the water, and the flow rate is obtained by cross-correlation flow rate measurement, which can determine the transit time of the flowing noise signals of fluid passing through the upstream and downstream sensors in the conductance sensor. In this article, the structure of the conductance sensor and the principles of measurement of flow rate and water cut are introduced. The results of the dynamic experiment in a multiphase flow loop and the in situ application of the technology are introduced herein.

Bz-423 superoxide signals B cell apoptosis via Mcl-1, Bak, and Bax

Bz-423 is a pro-apoptotic 1,4-benzodiazepine with therapeutic properties in murine models of lupus demonstrating selectivity for autoreactive lymphocytes. Bz-423 modulates the F 1F 0-ATPase, inducing the formation of superoxide within the mitochondrial respiratory chain, which then functions as a second messenger initiating apoptosis. In order to understand some of the features that contribute to the increased sensitivity of lymphocytes, we report the signaling pathway engaged by Bz-423 in a Burkitt lymphoma cell line (Ramos). Following the generation of superoxide, Bz-423-induced apoptosis requires the activation of Bax and Bak to induce mitochondrial outer membrane permeabilization and cytochrome c release. Knockdown of the BH3-only proteins Bad, Bim, Bik, and Puma inhibits Bz-423 apoptosis, suggesting that these proteins serve as upstream sensors of the oxidant stress induced by Bz-423. Treatment with Bz-423 results in superoxide-dependent Mcl-1 degradation, implicating this protein as the link between Bz-423-induced superoxide and Bax and Bak activation. In contrast to fibroblasts, B cell death induced by Bz-423 is independent of c-Jun N-terminal kinase. These results demonstrate that superoxide generated from the mitochondrial respiratory chain as a consequence of a respiratory transition can signal a specific apoptotic response that differs across cell types.

Insulation and Specificity in Bacterial Two‐Component Signaling Systems

Two‐component systems are one of the major modes of signal transduction in bacteria. In their simplest forms, these regulatory circuits are characterized by two proteins ‐ an upstream sensor kinase and a downstream response regulator. They mediate responses to diverse environmental signals and play a central role in regulating many aspects of microbial physiology. Two‐component circuits have been uncovered in remarkable numbers within individual organisms and across different bacterial species. This raises interesting questions regarding the mechanisms that these systems use to minimize inappropriate cross‐talk between homologous circuits. I will present recent work in which we have explored design aspects of the simplest examples of these circuits in E. coli. I will primarily focus on mechanisms that maintain signal strength and provide insulation against cross‐talk.

다결정 실리콘 카바이드를 이용한 마이크로 유량센서

A thermal flow sensor has been fabricated and characterized, consisting of a center resistive heater surrounded by two upstream and one downstream temperature sensing resistors. The heater and temperature sensing resistors are fabricated from nitrogen-doped(n-type) polycrystalline silicon carbide(poly-SiC) deposited by LPCVD(low pressure chemical vapor deposition) on LPCVD silicon nitride films on a Si substrate. Cavities were etched into the Si substrate from the front side to create suspended silicon nitride membranes carrying the poly-SiC elements. One upstream sensor is located <TEX>$50{\mu}m$</TEX> from the heater and has a sensitivity of <TEX>$0.73{\Omega}$</TEX>/sccm with <TEX>${\sim}15\;ms$</TEX> rise time in a dynamic range of 1000 sccm. N-type poly-SiC has a linear negative temperature coefficient and a TCR(temperature coefficient of resistance) of <TEX>$-1.24{\times}10^{-3}/^{\circ}C$</TEX> from room temperature to <TEX>$100^{\circ}C$</TEX>.

CARF Is a Vital Dual Regulator of Cellular Senescence and Apoptosis

The tumor suppressor protein, p53, is central to the pathways that monitor the stress, DNA damage repair, cell cycle, aging, and cancer. Highly complex p53 networks involving its upstream sensors and regulators, downstream effectors and regulatory feedback loops have been identified. CARF (Collaborator of ARF) was shown to enhance ARF-dependent and -independent wild-type p53 function. Here we report that (i) CARF overexpression causes premature senescence of human fibroblasts, (ii) it is vital for replicative and stress-induced senescence, and (iii) the lack of CARF function causes aneuploidy and apoptosis. We provide evidence that CARF plays a dual role in regulating p53-mediated senescence and apoptosis, the two major tumor suppressor mechanisms.

Optical tomography system using Digital Signal Processor: velocity profile measurement

The DSP system was customised to control the data acquisition process of 16 × 16 optical sensors array, arranged in parallel beam projection. The data collected was used to reconstruct the cross sectional image of the pipeline conveyor. For image display purpose, the reconstructed image was sent to a personal computer via serial communication. By measuring the flow velocity at each pixel, a profile of flow velocity is obtained and termed as Velocity Profile. The method used to measure the flow velocity at each pixel by cross correlating the upstream pixel values with respective downstream pixel values or upstream sensor values with respective downstream sensor values. The algorithm to perform cross correlation is discussed.

ArabidopsiseIF2α kinase GCN2 is essential for growth in stress conditions and is activated by wounding

BackgroundPhosphorylation of eIF2α provides a key mechanism for down-regulating protein synthesis in response to nutrient starvation or stresses in mammalian and yeast cells. However, this process has not been well characterized in plantsResultsWe show here that in response to amino acid and purine starvations, UV, cold shock and wounding, the Arabidopsis GCN2 kinase (AtGCN2) is activated and phosphorylates eIF2α. We show that AtGCN2 is essential for plant growth in stress situations and that its activity results in a strong reduction in global protein synthesis.ConclusionOur results suggest that a general amino acid control response is conserved between yeast and plants but that the plant enzyme evolved to fulfill a more general function as an upstream sensor and regulator of diverse stress-response pathways. The activation of AtGCN2 following wounding or exposure to methyl jasmonate, the ethylene precursor 1-Aminocyclopropane-1-carboxylic acid (ACC) and salicylic acid, further suggests that this enzyme could play a role in plant defense against insect herbivores.

Dual-Channel Pseudorandom Sequence Generator With Precise Time Delay Between Its Two Channels

In this paper, a dual-channel pseudorandom sequence generator is presented for the calibration of cross correlators in flow metering. In the sequence generator, the two channels have completely identical hardware structures so that they generate two identical sequences of pseudorandom signals to simulate two signals obtained from the upstream and downstream sensors of a cross correlator, respectively. Special control strategies in both hardware and software have been implemented in such a way that the error of time delay between the two channels can be reduced to less than a microsecond. The shift frequency and sequence length of each sequence channel and the time delay between the two channels can be adjusted in wide ranges to satisfy various requirements. The instrument has been evaluated by using a digital oscilloscope. Results obtained show that the control accuracy of time delay is equal to or better than 0.015% when the time delay is 1 ms or longer. Comparison with two commercially available noise signal generators shows that the error of time delay of the new generator is much smaller than those of the other two.

BAD: undertaker by night, candyman by day

The BH3-only pro-apoptotic proteins are upstream sensors of cellular damage that selectively respond to specific, proximal death and survival signals. Genetic models and biochemical studies indicate that these molecules are latent killers until activated through transcriptional or post-translational mechanisms in a tissue-restricted and signal-specific manner. The large number of BH3-only proteins, their unique subcellular localization, protein-interaction network and diverse modes of activation suggest specialization of their damage-sensing function, ensuring that the core apoptotic machinery is poised to receive input from a wide range of cellular stress signals. The apoptotic response initiated by the activation of BH3-only proteins ultimately culminates in allosteric activation of pro-apoptotic BAX and BAK, the gateway proteins to the mitochondrial pathway of apoptosis. From activation of BH3-only proteins to oligomerization of BAX and BAK and mitochondrial outer membrane permeabilization, an intricate network of interactions between the pro- and anti-apoptotic members of the BCL-2 family orchestrates the decision to undergo apoptosis. Beyond regulation of apoptosis, multiple BCL-2 proteins have recently emerged as active components of select homeostatic pathways carrying other cellular functions. This review focuses on BAD, which was the first BH3-only protein linked to proximal survival signals through phosphorylation by survival kinases. In addition to findings that delineated the physiological role of BAD in apoptosis and its dynamic regulation by phosphorylation, studies pointing to new roles for this protein in other physiological pathways, such as glucose metabolism, are highlighted. By executing its 'day' and 'night' jobs in metabolism and apoptosis, respectively, BAD helps coordinate mitochondrial fuel metabolism and the apoptotic machinery.