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An improved Geneva mechanism is characterized by a driven star-wheel having a segmented cam-follower surface. Star-wheel driver includes a restraining cam having a segmented cam surface for engaging the cam-follower surface of the star-wheel and antifriction rollers pinned to the restraining cam for engaging the cam-follower surface.
The regulatory protein, calmodulin (CaM), controls the activity of a plasma membrane localized ATPase in plants which serves to pump calcium out of cells. Recent data are consistent with the hypothesis that activation of this pump is one of the early steps necessary for gravitropism. Chlorpromazine (CPZ), a CaM antagonist, reversibly inhibits gravitropism in oat coleoptiles at concentrations which permit normal growth rates. C-14-labeled CPZ was used to photo-affinity label endogenous CaM in vivo to learn whether the drug is actually binding to some portion of endogenous CaM when it inhibits gravitropism. Under conditions in which CPZ inhibits gravitropism for over an hour, at least 11% of the CaM in gravitropically stimulated coleoptiles is bound to CPZ. In a given CPZ experiment the degree of inhibition of gravitropism correlates well with the amount of CaM bound to CPZ.
AS 2 part of the first joint USA-Russian MIR/Shuttle program, fertilized quail eggs were flown on the MIR 18 mission. Post-flight examination indicated impaired survival of both the embryos in space and also of control embryos exposed to vibrational and g-forces simulating the conditions experienced during the launch of Progress 227. We hypothesized that excess mechanical forces and/or other conditions during the launch might cause abnormal development of the blood supply in the chorioallantoic membrane (CAM) leading to the impaired survival of the embryos. The CAM, a highly vascularized extraembryonic organ, provides for the oxygen exchange across the egg shell and is thus pivotal for proper embryonic development. To test our hypothesis, we compared angiogenesis In CAMS of eggs which were either exposed to the vibration and g-force profile simulating the conditions at launch of Progress 227 (synchronous controls), or kept under routine conditions in a laboratory Incubator (laboratory controls). At various time points during Incubation, the eggs were fixed in paraformaldehyde for subsequent dissection. At the time of dissection, the CAM was carefully lifted from the egg shell and examined as whole mounts by bright-field and fluorescent microscopy. The development or the vasculature (angiogenesis) was assessed from the density of blood vessels per viewing field and evaluated by computer aided image analysis. We observed a significant decrease In blood-vessel density in the synchronous controls versus "normal" laboratory controls beginning from day 10 of Incubation. The decrease in vascular density was restricted to the smallest vessels only, suggesting that conditions during the launch and/or during the subsequent Incubation of the eggs may affect the normal progress of angiogenesis in the CAM. Abnormal angiogenesis In the CAM might contribute to the impaired survival of the embryos observed in synchronous controls as well as in space.
Calcium regulates diverse cellular activities in plants through the action of calmodulin (CaM). By using (35)S-labeled CaM to screen an Arabidopsis seedling cDNA expression library, a cDNA designated as AtCh-CPN10 (Arabidopsis thaliana chloroplast chaperonin 10) was cloned. Chloroplast CPN10, a nuclear-encoded protein, is a functional homolog of E. coli GroES. It is believed that CPN60 and CPN10 are involved in the assembly of Rubisco, a key enzyme involved in the photosynthetic pathway. Northern analysis revealed that AtCh-CPN10 is highly expressed in green tissues. The recombinant AtCh-CPN10 binds to CaM in a calcium-dependent manner. Deletion mutants revealed that there is only one CaM-binding site in the last 31 amino acids of the AtCh-CPN10 at the C-terminal end. The CaM-binding region in AtCh-CPN10 has higher homology to other chloroplast CPN10s in comparison to GroES and mitochondrial CPN10s, suggesting that CaM may only bind to chloroplast CPN10s. Furthermore, the results also suggest that the calcium/CaM messenger system is involved in regulating Rubisco assembly in the chloroplast, thereby influencing photosynthesis. Copyright 2000 Academic Press.
Ca(2+)-dependent protein kinases (CDPK) have a calmodulin-like domain (CaM-LD) tethered to the C-terminal end of the kinase. Activation is proposed to involve intramolecular binding of the CaM-LD to a junction sequence that connects the CaM-LD to the kinase domain. Consistent with this model, a truncated CDPK (DeltaNC) in which the CaM-LD has been deleted can be activated in a bimolecular interaction with an isolated CaM-LD or calmodulin, similar to the activation of a calmodulin-dependent protein kinase (CaMK) by calmodulin. Here we provide genetic evidence that this bimolecular activation requires a nine-residue binding segment from F436 to I444 (numbers correspond to CPK-1 accession number L14771). Two mutations at either end of this core segment (F436/A and VI444/AA) severely disrupted bimolecular activation, whereas flanking mutations had only minor effects. Intramolecular activation of a full-length kinase was also disrupted by a VI444/AA mutation, but surprisingly not by a F436/A mutation (at the N-terminal end of the binding site). Interestingly, intramolecular but not bimolecular activation was disrupted by insertion mutations placed immediately downstream of I444. To show that mutant enzymes were not misfolded, latent kinase activity was stimulated through binding of an antijunction antibody. Results here support a model of intramolecular activation in which the tether (A445 to G455) that connects the CaM-LD to the kinase provides an important structural constraint and is not just a simple flexible connection.
The Microgravity Research Program is part of NASA's Office of Biological and Physical Research (OBPR). The mission of the Microgravity Fluid Physics research program is to facilitate and conduct the best possible fluid physics research using the space environment and make this knowledge available to the scientific community and the public at large. During the summer of 2004, I worked in this division with Dr. Patricia Parsons-Wingerter. Dr. Parsons was working on several projects that used the chorioallantoic membrane (CAM) of Japanese quail eggs. The CAM develops in the eggs of birds and reptiles and is a very vascular fetal membrane composed of the fused chorion and adjacent wall of the allantois. The CAM is formed on day 4 of incubation and its primary job is to mediate gas exchanges with the extra embryonic environment. The CAM of our Japanese quail eggs is easily identifiable to us because it is transparent and it sits on top of the yolk with the embryo in the center. The CAM is of interest because of its many applications in the field of medicine as it relates to vascular remodeling and angiogenesis. Angiogenesis is simply the growth or formation of new blood vessels and anti-angiogenesis is the inhibition of said vessels. Angiogenesis occurs naturally in a healthy body for healing wounds and for restoring blood flow to tissues after injury and in females during the monthly reproductive cycle. In many serious diseases, like several types of cancer and those that affect the heart and cardiovascular system, the body loses control over angiogenesis. These diseases, which are dependent on angiogenesis, result when new blood vessels either grow excessively or insufficiently. The chorioallantoic membrane of our Japanese quail eggs gives a good model of angiogenesis. We used angiogenic regulators to inhibit or stimulate vascular growth in the CAM in a healthy manner and they induced distinct vascular patterns in vivo. Certain dominant regulators can be recognized by their unique vascular patterns and from these patterns; we can deduce specific alterations in vascular remodeling and angiogenesis. This will aid us in early-stage diagnosis and customized therapies for patients with angiogenic-dependent diseases. This particular research is important to NASA because cardiovascular health issues are the second highest of ten categories that have been defined as risk factors in human space exploration. Also, cardiovascular-related diseases have been the leading cause of death in America since 1981. Therefore, this kind of research in the field of cardiovascular health is of great importance to humans on earth and in space. This particular research is important to NASA because cardiovascular health is of great importance to humans on earth and in space.
Abstract Antimony‐119 ( 119 Sb) is one of the most attractive Auger‐electron emitters identified to date, but it remains practically unexplored for targeted radiotherapy because no chelators have been identified to stably bind this metalloid in vivo. In a departure from current studies focused on chelator development for Sb(III), we explore the chelation chemistry of Sb(V) using the tris‐catecholate ligand TREN‐CAM. Through a combination of radiolabeling, spectroscopic, solid‐state, and computational studies, the radiochemistry and structural chemistry of TREN‐CAM with 1XX/nat Sb(V) were established. The resulting [ 1XX Sb]Sb–TREN‐CAM complex remained intact for several days in human serum, signifying high stability under biological conditions. Finally, the first in vivo single photon emission computed tomography and positron emission tomography imaging studies were carried out using 117 Sb, the diagnostic analogue of 119 Sb. These studies revealed marked differences in the uptake and distribution of activity in mice administered unchelated [ 117 Sb]Sb(OH) 6 – versus [ 117 Sb]Sb–TREN‐CAM, suggesting that 117 Sb is largely retained by TREN‐CAM over the time course of the study. Collectively, these findings demonstrate the most physiologically stable complex of no‐carrier‐added 1XX Sb yet reported, offering new promise for the clinical implementation of radioantimony in nuclear medicine. Our results also establish the feasibility of 117 Sb as an elementally matched partner to 119 Sb for theranostic applications.
Antimony-119 ( 119 Sb) is one of the most attractive Auger-electron emitters identified to date, but it remains practically unexplored for targeted radiotherapy because no chelators have been identified to stably bind this metalloid in vivo. In a departure from current studies focused on chelator development for Sb(III), we explore the chelation chemistry of Sb(V) using the tris-catecholate ligand TREN-CAM. Through a combination of radiolabeling, spectroscopic, solid-state, and computational studies, the radiochemistry and structural chemistry of TREN-CAM with 1XX/nat Sb(V) were established. The resulting [ 1XX Sb]Sb–TREN-CAM complex remained intact for several days in human serum, signifying high stability under biological conditions. Finally, the first in vivo single photon emission computed tomography and positron emission tomography imaging studies were carried out using 117 Sb, the diagnostic analogue of 119 Sb. These studies revealed marked differences in the uptake and distribution of activity in mice administered unchelated [ 117 Sb]Sb(OH) 6 – versus [ 117 Sb]Sb–TREN-CAM, suggesting that 117 Sb is largely retained by TREN-CAM over the time course of the study. Collectively, these findings demonstrate the most physiologically stable complex of no-carrier-added 1XX Sb yet reported, offering new promise for the clinical implementation of radioantimony in nuclear medicine. In conclusion, our results also establish the feasibility of 117 Sb as an elementally matched partner to 119 Sb for theranostic applications.
An electromagnetic brake/clutch device includes a drive shaft supported by at least one bearing for transmitting torque, a housing, affixed to prevent its rotation, surrounding the drive shaft, and an electromagnetically activated device within the housing to selectively prevent and allow rotation of the drive shaft. The electromagnetically activated device includes a plurality of cammed rollers to prevent counter-clockwise rotation of the drive shaft. The drive shaft includes a circumferential disk and the housing includes a reaction ring for engagement with the plurality of cammed rollers. The plurality of cammed rollers are released from engagement with the circumferential disk and the reaction ring by a plurality of tripping mechanisms within the housing. The tripping action uses the locking force to act as a release force merely by changing the boundary conditions of the roller interface angles. The tripping mechanisms include trippers for disengaging the plurality of cammed rollers and an anvil shaped portion for providing lateral movement of the trippers. The plurality of cammed rollers is preloaded to engagement with the circumferential disk and reaction ring by a spring, and is located with respect to an adjacent tripping mechanism with another spring.
Calmodulin (CaM) was used as an affinity tail to facilitate the purification of the green fluorescent protein (GFP), which was used as a model target protein. The protein GFP was fused to the C-terminus of CaM, and a factor Xa cleavage site was introduced between the two proteins. A CaM-GFP fusion protein was expressed in E. coli and purified on a phenothiazine-derivatized silica column. CaM binds to the phenothiazine on the column in a Ca(2+)-dependent fashion and it was, therefore, used as an affinity tail for the purification of GFP. The fusion protein bound to the affinity column was then subjected to a proteolytic digestion with factor Xa. Pure GFP was eluted with a Ca(2+)-containing buffer, while CaM was eluted later with a buffer containing the Ca(2+)-chelating agent EGTA. The purity of the isolated GFP was verified by SDS-PAGE, and the fluorescence properties of the purified GFP were characterized.
Cell adhesion molecules (CAMs) are proteins which anchor cells to each other and to the extracellular matrix (ECM), but whose functions also include signal transduction, differentiation, and apoptosis. We are testing a hypothesis that particle radiations modulate CAM expression and this contributes to radiation-induced lens opacification. We observed dose-dependent changes in the expression of beta 1-integrin and ICAM-1 in exponentially-growing and confluent cells of a differentiating human lens epithelial cell model after exposure to particle beams. Human lens epithelial (HLE) cells, less than 10 passages after their initial culture from fetal tissue, were grown on bovine corneal endothelial cell-derived ECM in medium containing 15% fetal bovine serum and supplemented with 5 ng/ml basic fibroblast growth factor (FGF-2). Multiple cell populations at three different stages of differentiation were prepared for experiment: cells in exponential growth, and cells at 5 and 10 days post-confluence. The differentiation status of cells was characterized morphologically by digital image analysis, and biochemically by Western blotting using lens epithelial and fiber cell-specific markers. Cultures were irradiated with single doses (4, 8 or 12 Gy) of 55 MeV protons and, along with unirradiated control samples, were fixed using -20 degrees C methanol at 6 hours after exposure. Replicate experiments and similar experiments with helium ions are in progress. The intracellular localization of beta 1-integrin and ICAM-1 was detected by immunofluorescence using monoclonal antibodies specific for each CAM. Cells known to express each CAM were also processed as positive controls. Both exponentially-growing and confluent, differentiating cells demonstrated a dramatic proton-dose-dependent modulation (upregulation for exponential cells, downregulation for confluent cells) and a change in the intracellular distribution of the beta 1-integrin, compared to unirradiated controls. In contrast, there was a dose-dependent increase in ICAM-1 immunofluorescence in confluent, but not exponentially-growing cells. These results suggest that proton irradiation downregulates beta 1-integrin and upregulates ICAM-1, potentially contributing to cell death or to aberrant differentiation via modulation of anchorage and/or signal transduction functions. Quantification of the expression levels of the CAMs by Western analysis is in progress.
We examined the effect of calmodulin (CaM) antagonists applied at the root tip on root growth, gravity-induced root curvature, and the movement of calcium across the root tip and auxin (IAA) across the elongation zone of gravistimulated roots. All of the CaM antagonists used in these studies delayed gravity-induced curvature at a concentration (1 micromole) that did not affect root growth. Calmodulin antagonists (> or = 1 micromole) inhibited downward transport of label from 45Ca2+ across the caps of gravistimulated roots relative to the downward transport of 45Ca2+ in gravistimulated roots which were not treated with CaM antagonists. Application of CaM antagonists at the root tip (> or = 1 micromole) also decreased the relative downward movement of label from 3H-IAA applied to the upper side of the elongation zone of gravistimulated roots. In general, tip application of antagonists inhibited neither the upward transport of 45Ca2+ in the root tip nor the upward movement of label from 3H-IAA in the elongation zone of gravistimulated roots. Thus, roots treated with CaM antagonists > or = 1 micromole become less graviresponsive and exhibit reduced or even a reversal of downward polarity of calcium transport across the root tip and IAA transport across the elongation zone. The results indicate that calmodulin-regulated events play a role in root gravitropism.
Objective: GeoCarb validation •Purpose: •Do the four TCCON sites in the GeoCarb domain characterize satellite validation? •Method: •Assess the error estimates globally vs. in the GeoCarb domain •Assess the use of GML/DOE aircraft for CH4 validation •Systematic error estimate (from Kulawik, 2016): •+ regional bias (stdevof average bias versus validation at each station) •+ correlated error (stdev of daily average versus validation) •-co-location error (above quantities for model@satelliteminus model@valid) •-validation error (systematic error of validation estimates, e.g. model extension) Aircraft validation error •CH4 profile extension error estimate: 7 ppb •Extend aircraft observations to the top of the atmosphere with different model configurations •CAMS, scaling to stratosphere, then taking stratosphere •Extend by repeating top aircraft value to the troposphere, then taking CAMS •Extend with GEOS-Chem run (from 2010). •Difference CAMS-Scale vs. CAMS-Tropo: •3.6 +-5.5 ppb •Systematic error: 6.6 ppb •Extending aircraft to tropopause is blatantly wrong, but these two have the same stratosphere. •Difference CAMS-Scale vs. GEOS-Scale: •6.2 +-8.7 ppb •Do not consider bias since model is 10 years off. Systematic error ~8.7 ppb. •Estimate of systematic error from profile extension (possibly underestimated, resulting in possibly higher TROPOMI error estimate) •7 ppb •Would likely vary some by season, site, other conditions •Aircraft measurement error: 1 ppb •1 ppb from measurement error3
The transition to electric vehicles (EVs) is pivotal for achieving energy security and integrating grid stability, with lithium-ion batteries (LIBs) playing a central role in this transformation. However, the conventional wet electrode manufacturing relying on N-methyl-2-pyrrolidone (NMP) solvent is energy intensive and costly. Dry processing (DP) has emerged as a promising alternative, eliminating solvents and using polytetrafluoroethylene (PTFE) binder for electrode fabrications. Despite its advantages, DP faces a critical challenge: poor interfacial adhesion between the hydrophobic PTFE binder and the hydrophilic cathode active material (CAM), particularly LiNi 0.8 Mn 0.1 Co 0.1 O 2 (NMC811), which undermines electrode performance. Here, to address this, we introduced a novel vapor-phase trimethoxymethylsilane (TMMS) coating to hydrophobize the CAM surface, enhancing compatibility with PTFE. This surface modification significantly enhances binder – CAM interactions, enabling uniform mixing and robust electrode integrity without damaging the CAM particles. Our findings advance the feasibility of environmentally sustainable and cost-effective dry processing, representing a significant step toward sustainable battery manufacturing.
All-solid-state lithium batteries (ASSLBs) are emerging as promising next-generation batteries for electric vehicles owing to their high energy densities and safety features. However, challenges such as inadequate material percolation and low cathode utilization often hinder their potential. This paper presents a core–shell approach to optimize the cathode active material (CAM) utilization. The resultant CAM composite showed high ionic conductivity, a highly dense microstructure with <10% porosity, and minimal stack pressure changes during electrochemical cycling. The maximum CAM utilization was achieved while effectively mitigating electrochemo-mechanical side reactions by applying a uniformly coated Li 6 PS 5 Cl solid electrolyte layer (≈500 nm) and a LiNbO 3 buffer layer (≈10 nm) onto LiNi 0.8 Mn 0.1 Co 0.1 O 2 particles (LPSCl@LNO@NMC). The engineered LPSCl@LNO@NMC composites, which incorporated a 5 wt% LPSCl coating on LNO@NMC powders, exhibited a dense microstructure that enhanced the mechanical stability at the cathode. Sulfide-based solid electrolyte (SSE)/SSE contact provided better ionic pathways within the composite and increased CAM utilization. Thus, an enhanced reversible capacity (197 mA h g -1 ) and exceptional high-rate cycling performance (86.3% capacity retention after 1000 cycles at 2C) were observed. These findings pave the way for the advancement and commercialization of high-performance ASSLBs.
Recently, gahnite (ZnAl 2 O 4 ) is gaining attraction as a potential refractory ceramic because of the similarity of its structure and properties with those of magnesium aluminate (MgAl 2 O 4 ) spinel refractories. Formation of MgO and hibonite solid solution (CaMg x Al 12−x O 19−0.5x ; 0 ≤ x ≤ 0.18), CAM-I (Ca 2 Mg 2−3x Al 28+2x O 46 (0 ≤ x ≤ 0.3), and CAM-II (CaMg 2−3x Al 16+2x O 27 , 0 ≤ x ≤ 0.2) phases with platelet and interlocking microstructure in the CaO–Al 2 O 3 –MgO ternary system significantly enhances the high temperature mechanical properties of refractory castables. The CaO–Al 2 O 3 –ZnO ternary system has been studied, for the first time to our knowledge, in a selected compositional range with reference to the CaO–Al 2 O 3 –MgO system from 1650°C to 1700°C. The formation of ZnO and hibonite solid solution (CaZn x Al 12−x O 19−0.5x ; 0 < x < 0.18), CAZ-I (Ca 2 Zn 2−3x Al 28+2x O 46 ; 0 ≤ x ≤ 0.3), and CAZ-II (CaZn 2−3x Al 16+2x O 27 ; 0 ≤ x ≤ 0.2) phases with platelet and interlocking morphology have been found. The crystal structures and lattice parameters of ZnO and hibonite solid solution, CAZ-I, and CAZ-II are comparable, respectively, with MgO and hibonite solid solution, CAM-I, and CAM-II. Furthermore, CAZ-I and CAZ-II phases also form due to reaction between hibonite (CaO·6Al 2 O 3 ) and ZnAl 2 O 4 .
Chronic hepatitis B virus (HBV) infection is strongly associated with increased risk of liver cancer and cirrhosis. While existing treatments effectively inhibit the HBV life cycle, viral rebound frequently occurs following treatment interruption. Consequently, functional cure rates of chronic HBV infection remain low and there is increased interest in a novel treatment modality, capsid assembly modulators (CAMs). Here, we develop a multiscale mathematical model of CAM treatment in chronic HBV infection. By fitting the model to participant data from a phase I trial of the first-generation CAM vebicorvir, we estimate the drug’s dose-dependent effectiveness and identify the physiological mechanisms that drive the observed biphasic decline in HBV DNA and RNA, and mechanistic differences between HBeAg-positive and negative infection. Finally, we demonstrate analytically and numerically that the relative change of HBV RNA more accurately reflects the antiviral effectiveness of a CAM than the relative change in HBV DNA.
Estimates of aviation effective radiative forcing (ERF) indicate that contrail cirrus is currently its largest contributor, although with a substantial associated uncertainty of ∼ 70 %. Here, we implement the contrail parameterisation developed for the Community Atmosphere Model (CAM) in the UK Met Office Unified Model (UM), allowing us to compare, for the first time, the impact of key features of the host climate model on contrail cirrus ERF. We find that differences in background humidity between the models result in the UM-simulated contrail fractions being 2 to 3 times larger than in CAM. Additionally, the models show contrasting responses in overall global cloud fraction, with contrails increasing the total cloud fraction in the UM and decreasing it in CAM. Differences in the complexity of the cloud microphysics schemes lead to significant differences in simulated changes to cloud ice water content due to aviation. After compensating for the unrealistically low contrail optical depth in the UM, we estimate the 2018 contrail cirrus ERF to be 40.8 mW m−2 in the UM, compared to 60.1 mW m−2 in CAM. These values highlight the substantial uncertainty in contrail cirrus ERF due to differences in microphysics and radiation schemes between the two models. We also find a factor-of-8 uncertainty in contrail cirrus ERF due to existing uncertainty in contrail cirrus optical depth. Future research should focus on better representing microphysical and radiative contrail characteristics in climate models and on improved observational constraints.