Imbalances in endoplasmic reticulum (ER) proteostasis are associated with etiologically-diverse degenerative diseases linked to excessive extracellular protein misfolding and aggregation. ...Reprogramming of the ER proteostasis environment through genetic activation of the Unfolded Protein Response (UPR)-associated transcription factor ATF6 attenuates secretion and extracellular aggregation of amyloidogenic proteins. Here, we employed a screening approach that included complementary arm-specific UPR reporters and medium-throughput transcriptional profiling to identify non-toxic small molecules that phenocopy the ATF6-mediated reprogramming of the ER proteostasis environment. The ER reprogramming afforded by our molecules requires activation of endogenous ATF6 and occurs independent of global ER stress. Furthermore, our molecules phenocopy the ability of genetic ATF6 activation to selectively reduce secretion and extracellular aggregation of amyloidogenic proteins. These results show that small molecule-dependent ER reprogramming, achieved through preferential activation of the ATF6 transcriptional program, is a promising strategy to ameliorate imbalances in ER function associated with degenerative protein aggregation diseases.
The mammalian heart pumps blood through the vessels, maintaining the dynamic equilibrium in a circulatory system driven by two pumps in series. This vital function is based on the fine-tuning of ...cardiac performance by the Frank–Starling mechanism that relates the pressure exerted by the contracting ventricle (end systolic pressure) to its volume (end systolic volume). At the level of the sarcomere, the structural unit of the cardiac myocytes, the Frank–Starling mechanism consists of the increase in active forcewith the increase of sarcomere length (length-dependent activation). We combine sarcomere mechanics and micrometer–nanometer-scale X-ray diffraction from synchrotron light in intact ventricular trabeculae from the rat to measure the axial movement of the myosin motors during the diastole–systole cycle under sarcomere length control. We find that the number of myosin motors leaving the off, ATP hydrolysis-unavailable state characteristic of the diastole is adjusted to the sarcomere length-dependent systolic force. This mechanosensing-based regulation of the thick filament makes the energetic cost of the systole rapidly tuned to the mechanical task, revealing a prime aspect of the Frank–Starling mechanism. The regulation is putatively impaired by cardiomyopathycausing mutations that affect the intramolecular and intermolecular interactions controlling the off state of the motors.
C-C motif chemokine receptor-like 2 (CCRL2) is a non-signaling 7 transmembrane receptor that binds chemotactic ligands to shape leukocyte recruitment to sites of inflammation. However, there is a ...lack of consensus on the ligands that directly bind CCRL2 or their functional impact. Studies with CCRL2 knockout mice have demonstrated that neutrophils have impaired degranulation and migration in response to CXCL8, where the underlying molecular mechanism is proposed to be due to the formation of CCRL2 heterodimers with the chemokine receptor CXCR2. Herein, we characterized the ligands that bind directly to CCRL2 and interrogated the impact of CCRL2 neutralization on CXCL8 signaling in neutrophils using pharmacological antibody tools. Using flow cytometry and Surface Plasmon Resonance microscopy (SPRm) cell binding experiments, we confirmed that chemerin, but not previously reported C-C chemokines, binds CCRL2. Furthermore, we identified human and mouse CCRL2 antibodies that neutralized chemerin binding to CCRL2. Unexpectedly, we found that neutralization of CCRL2 with these antibodies did not attenuate CXCL8-induced human neutrophil degranulation nor CXCL8-induced murine neutrophil recruitment to the peritoneum. Based on the observed differences in modulating CCRL2 function with neutralizing antibodies compared to the reported CCRL2 deficient murine models, we hypothesize that the ligand binding function of CCRL2 is dispensable for CXCL8 signaling in neutrophils. Finally, extensive profiling of CCRL2 expression on peripheral blood leukocytes revealed monocytes, dendritic cells (DC), and subpopulations of natural killer T (NKT) cells as additional targets, highlighting potential roles for CCRL2 in human cell types beyond neutrophils that warrants future investigation.
Predicting future impacts of anthropogenic change on tropical forests requires a clear understanding of nutrient constraints on productivity. We compared experimental fertilization and litter ...manipulation treatments in an old-growth lowland tropical forest to distinguish between the effects of inorganic nutrient amendments and changes in nutrient cycling via litterfall. We measured the changes in soil and litter nutrient pools, litterfall, and fine root biomass in plots fertilized with nitrogen (N), phosphorus (P), or potassium (K), and in litter addition and litter removal treatments during 7 years. Soil inorganic N and litter N increased in double-litter plots but not in N-fertilized plots. Conversely, litter P and soil pools of P and increased in fertilized plots but not in the double-litter plots. Soil and litter pools of N and K decreased in the no-litter plots. Changes in litterfall with added nutrients or litter were only marginally significant, but fine root biomass decreased with both the litter and the K addition. Differences between the two experiments are mostly attributable to the coupled cycling of carbon and nutrients in litter. Increased nutrient inputs in litter may improve plant uptake of some nutrients compared to fertilization with similar amounts. The litter layer also appears to play a key role in nutrient retention. We discuss our findings in the context of possible impacts of anthropogenic change on tropical forests.
Lianas in tropical forests compete intensely with trees for above- and belowground resources and limit tree growth and regeneration. Liana competition with adult canopy trees may be particularly ...strong, and, if lianas compete more intensely with some tree species than others, they may influence tree species composition. We performed the first systematic, large-scale liana removal experiment to assess the competitive effects of lianas on multiple tropical tree species by measuring sap velocity and growth in a lowland tropical forest in Panama. Tree sap velocity increased 60% soon after liana removal compared to control trees, and tree diameter growth increased 25% after one year. Although tree species varied in their response to lianas, this variation was not significant, suggesting that lianas competed similarly with all tree species examined. The effect of lianas on tree sap velocity was particularly strong during the dry season, when soil moisture was low, suggesting that lianas compete intensely with trees for water. Under the predicted global change scenario of increased temperature and drought intensity, competition from lianas may become more prevalent in seasonal tropical forests, which, according to our data, should have a negative effect on most tropical tree species.
Translucent zirconias have been developed with better esthetics than high-strength zirconias by reducing opacity. However, studies on their translucency and strength are sparse.
The purpose of this ...in vitro study was to compare the relationship between translucency and biaxial flexural strength of recently developed high-translucency zirconia, high-strength zirconia, and lithium disilicate ceramics.
Disks (n=12) were fabricated for 5 ceramic materials: high-strength zirconia (BruxZir 16 shaded), translucent zirconia (BruxZir Anterior shaded, Katana UTML, Katana STML), and lithium disilicate (IPS e.max, Press HT, and LT). A standard tessellation language (STL) file was designed, and the specimen milled, finished, and glazed according to manufacturer's instructions for each material. The translucency parameter was calculated against black and white backgrounds and white and stump shade with ND4 background by using a spectrophotometer. Biaxial flexural strength was calculated by using the 3-ball test. The load was applied at a crosshead speed of 0.5 mm/min with a 49-N load cell until failure occurred. Translucency parameter and biaxial flexural strength data were analyzed with 1-way analysis of variance (ANOVA). A Tukey honest significant difference multiple comparison test was used to determine significant differences (α=.05).
The IPS e.max HT was more translucent against both backgrounds (32.85 for black/white and 15.34 for white/stump), while BruxZir 16 was the least translucent (19.78 for B/W and 8.83 for W/S). All groups tested differed in translucency (P<.001) except for BruxZir Anterior and Katana STML, which were not significantly different (P=.052). For biaxial flexural strength, BruxZir 16 had the highest strength (995.44 MPa) and e.max HT, the lowest (186.75 MPa). No significant differences were found between BruxZir anterior and Katana STML, Katana UTML and IPS e.max LT, or IPS e.max LT and IPS e.max HT (P>.05). Translucency parameter values using both backgrounds were strongly correlated (r=0.99). However, biaxial flexural strength values were inversely related to translucency parameter values when using black/white and white/stump shade but with high correlation (r=-0.777 and -0.756 respectively).
Lithium disilicate was the most translucent and yet the weakest material, whereas high-strength zirconia was the most opaque ceramic and the strongest. Katana UTML had the highest translucency but was weakest among translucent and high-strength zirconia materials. Overall, translucency was negatively correlated with biaxial flexural strength.
Nematodes are the most abundant and diverse metazoans on Earth, and are known to significantly affect ecosystem functioning. A better understanding of their biology and ecology, including potential ...adaptations to diverse habitats and lifestyles, is key to understanding their response to global change scenarios. Mitochondrial genomes offer high species level characterization, low cost of sequencing, and an ease of data handling that can provide insights into nematode evolutionary pressures. Generally, nematode mitochondrial genomes exhibited similar structural characteristics (e.g., gene size and GC content), but displayed remarkable variability around these general patterns. Compositional strand biases showed strong codon position specific G skews and relationships with nematode life traits (especially parasitic feeding habits) equal to or greater than with predicted phylogeny. On average, nematode mitochondrial genomes showed low non-synonymous substitution rates, but also high clade specific deviations from these means. Despite the presence of significant mutational saturation, non-synonymous (dN) and synonymous (dS) substitution rates could still be significantly explained by feeding habit and/or habitat. Low ratios of dN:dS rates, particularly associated with the parasitic lifestyles, suggested the presence of strong purifying selection. Nematode mitochondrial genomes demonstrated a capacity to accumulate diversity in composition, structure, and content while still maintaining functional genes. Moreover, they demonstrated a capacity for rapid evolutionary change pointing to a potential interaction between multi-level selection pressures and rapid evolution. In conclusion, this study helps establish a background for our understanding of the potential evolutionary pressures shaping nematode mitochondrial genomes, while outlining likely routes of future inquiry.
Key points
Myosin filament mechanosensing determines the efficiency of the contraction by adapting the number of switched ON motors to the load.
Accordingly, the unloaded shortening velocity (V0) is ...already set at the end of latency relaxation (LR), ∼10 ms after the start of stimulation, when the myosin filament is still in the OFF state.
Here the number of actin‐attached motors per half‐myosin filament (n) during V0 shortening imposed either at the end of LR or at the plateau of the isometric contraction is estimated from the relation between half‐sarcomere compliance and force during the force redevelopment after shortening.
The value of n decreases progressively with shortening and, during V0 shortening starting at the end of LR, is 1–4.
Reduction of n is accounted for by a constant duty ratio of 0.05 and a parallel switching OFF of motors, explaining the very low rate of ATP utilization found during unloaded shortening.
The maximum velocity at which a skeletal muscle can shorten (i.e. the velocity of sliding between the myosin filament and the actin filament under zero load, V0) is already set at the end of the latency relaxation (LR) preceding isometric force generation, ∼10 ms after the start of electrical stimulation in frog muscle fibres at 4°C. At this time, Ca2+‐induced activation of the actin filament is maximal, while the myosin filament is in the OFF state characterized by most of the myosin motors lying on helical tracks on the filament surface, making them unavailable for actin binding and ATP hydrolysis. Here, the number of actin‐attached motors per half‐thick filament during V0 shortening (n) is estimated by imposing, on tetanized single fibres from Rana esculenta (at 4°C and sarcomere length 2.15 μm), small 4 kHz oscillations and determining the relation between half‐sarcomere (hs) compliance and force during the force development following V0 shortening. When V0 shortening is superimposed on the maximum isometric force T0, n decreases progressively with the increase of shortening (range 30–80 nm per hs) and, when V0 shortening is imposed at the end of LR, n can be as low as 1–4. Reduction of n is accounted for by a constant duty ratio of the myosin motor of ∼0.05 and a parallel switching OFF of the thick filament, providing an explanation for the very low rate of ATP utilization during extended V0 shortening.
Key points
Myosin filament mechanosensing determines the efficiency of the contraction by adapting the number of switched ON motors to the load.
Accordingly, the unloaded shortening velocity (V0) is already set at the end of latency relaxation (LR), ∼10 ms after the start of stimulation, when the myosin filament is still in the OFF state.
Here the number of actin‐attached motors per half‐myosin filament (n) during V0 shortening imposed either at the end of LR or at the plateau of the isometric contraction is estimated from the relation between half‐sarcomere compliance and force during the force redevelopment after shortening.
The value of n decreases progressively with shortening and, during V0 shortening starting at the end of LR, is 1–4.
Reduction of n is accounted for by a constant duty ratio of 0.05 and a parallel switching OFF of motors, explaining the very low rate of ATP utilization found during unloaded shortening.
► We propose a novel rapid method using infrared spectroscopy to quantify total antioxidant capacity in plants in the substitution of traditional wet chemistry methodology. ► Several ...spectroscopic-based chemometric models were established and validated. Those models could be used in future for prediction of onion antioxidant profile. ► DPPH, TEAC and FRAP values of different types of onions and shallots were measured and those data are partially supplemented to previous relevant studies.
Total phenolic content (TPC) and total antioxidant capacity (TAC) of four onion varieties (red, white, yellow and sweet) and shallot from selected locations (Washington, Idaho, Oregon, Texas and Georgia) were determined using Fourier transform infrared (FT-IR) spectroscopy (4000–400cm−1). The Folin–Ciocalteu (F–C) assay was used to quantify TPC and three assays were used to determine TAC, including 2,2-diphenyl-picrylhydrazyl (DPPH) assay, Trolox equivalent antioxidant capacity (TEAC) assay and ferric reducing antioxidant power (FRAP) assay. Partial least squares regression (PLSR) with cross-validation (leave-one-out) was conducted on onion and shallot extracts (n=200) and their corresponding F–C, DPPH, TEAC and FRAP values were employed to obtain four independent calibration models for predicting TPC and TAC for the extracts. Spectra from an extra 19 independent extracts were used as an external validation set for prediction. A correlation of r>0.95 was obtained between FT-IR predicted and reference values (by F–C, DPPH, TEAC and FRAP assay) with standard errors of calibration (SEC) and standard errors of cross-validation (SECV) less than 2.85, 0.35 and 0.45μmolTrolox/g FW of extracts for TEAC, FRAP and DPPH assay, respectively; and 0.36mggallic acid/g FW of extracts for the F–C assay. In addition, cluster analysis (principal component analysis (PCA)) and discriminant function analysis (DFA) could differentiate varieties of onions and shallot based upon infrared spectral features. Loading plots for the various chemometrics models indicated that hydroxyl and phenolic functional groups were most closely correlated with antioxidant capacity. The use of mid-infrared spectroscopy to predict the total antioxidant capacity of vegetables provides a rapid and precise alternative to traditional wet chemistry analysis.
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