Neurodegenerative lysosomal storage disorders (LSDs) are severe and untreatable, and mechanisms underlying cellular dysfunction are poorly understood. We found that toxic lipids relevant to three ...different LSDs disrupt multiple lysosomal and other cellular functions. Unbiased drug discovery revealed several structurally distinct protective compounds, approved for other uses, that prevent lysosomal and cellular toxicities of these lipids. Toxic lipids and protective agents show unexpected convergence on control of lysosomal pH and re-acidification as a critical component of toxicity and protection. In twitcher mice (a model of Krabbe disease KD), a central nervous system (CNS)-penetrant protective agent rescued myelin and oligodendrocyte (OL) progenitors, improved motor behavior, and extended lifespan. Our studies reveal shared principles relevant to several LSDs, in which diverse cellular and biochemical disruptions appear to be secondary to disruption of lysosomal pH regulation by specific lipids. These studies also provide novel protective strategies that confer therapeutic benefits in a mouse model of a severe LSD.
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DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Purines and related heterocycles substituted at C-2 with 4′-sulfamoylanilino and at C-6 with a variety of groups have been synthesized with the aim of achieving selectivity of binding to CDK2 over ...CDK1. 6-Substituents that favor competitive inhibition at the ATP binding site of CDK2 were identified and typically exhibited 10–80-fold greater inhibition of CDK2 compared to CDK1. Most impressive was 4-((6-(1,1′-biphenyl-3-yl)-9H-purin-2-yl)amino) benzenesulfonamide (73) that exhibited high potency toward CDK2 (IC50 0.044 μM) but was ∼2000-fold less active toward CDK1 (IC50 86 μM). This compound is therefore a useful tool for studies of cell cycle regulation. Crystal structures of inhibitor–kinase complexes showed that the inhibitor stabilizes a glycine-rich loop conformation that shapes the ATP ribose binding pocket and that is preferred in CDK2 but has not been observed in CDK1. This aspect of the active site may be exploited for the design of inhibitors that distinguish between CDK1 and CDK2.
Cancer-associated fibroblasts (CAFs) are tumor-promoting and correlate with poor survival in many cancers, which has led to their emergence as potential therapeutic targets. However, effective ...methods to manipulate these cells clinically have yet to be developed.
CAF accumulation and prognostic significance in head and neck cancer (oral, n = 260; oropharyngeal, n = 271), and colorectal cancer (n = 56) was analyzed using immunohistochemistry. Mechanisms regulating fibroblast-to-myofibroblast transdifferentiation were investigated in vitro using RNA interference/pharmacological inhibitors followed by polymerase chain reaction (PCR), immunoblotting, immunofluorescence, and functional assays. RNA sequencing/bioinformatics and immunohistochemistry were used to analyze NAD(P)H Oxidase-4 (NOX4) expression in different human tumors. NOX4's role in CAF-mediated tumor progression was assessed in vitro, using CAFs from multiple tissues in Transwell and organotypic culture assays, and in vivo, using xenograft (n = 9-15 per group) and isograft (n = 6 per group) tumor models. All statistical tests were two-sided.
Patients with moderate/high levels of myofibroblastic-CAF had a statistically significant decrease in cancer-specific survival rates in each cancer type analyzed (hazard ratios HRs = 1.69-7.25, 95% confidence intervals CIs = 1.11 to 31.30, log-rank P ≤ .01). Fibroblast-to-myofibroblast transdifferentiation was dependent on a delayed phase of intracellular reactive oxygen species, generated by NOX4, across different anatomical sites and differentiation stimuli. A statistically significant upregulation of NOX4 expression was found in multiple human cancers (P < .001), strongly correlating with myofibroblastic-CAFs (r = 0.65-0.91, adjusted P < .001). Genetic/pharmacological inhibition of NOX4 was found to revert the myofibroblastic-CAF phenotype ex vivo (54.3% decrease in α-smooth muscle actin α-SMA, 95% CI = 10.6% to 80.9%, P = .009), prevent myofibroblastic-CAF accumulation in vivo (53.2%-79.0% decrease in α-SMA across different models, P ≤ .02) and slow tumor growth (30.6%-64.0% decrease across different models, P ≤ .04).
These data suggest that pharmacological inhibition of NOX4 may have broad applicability for stromal targeting across cancer types.
Room-temperature ionic liquids (RTILs) are nonvolatile, tunable solvents that have generated significant interest across a wide variety of engineering applications. The use of RTILs as media for CO2 ...separations appears especially promising, with imidazolium-based salts at the center of this research effort. The solubilities of gases, particularly CO2, N2, and CH4, have been studied in a number of RTILs. Process temperature and the chemical structures of the cation and anion have significant impacts on gas solubility and gas pair selectivity. Models based on regular solution theory and group contributions are useful to predict and explain CO2 solubility and selectivity in imidazolium-based RTILs. In addition to their role as a physical solvent, RTILs might also be used in supported ionic liquid membranes (SILMs) as a highly permeable and selective transport medium. Performance data for SILMs indicates that they exhibit large permeabilities as well as CO2/N2 selectivities that outperform many polymer membranes. Furthermore, the greatest potential of RTILs for CO2 separations might lie in their ability to chemically capture CO2 when used in combination with amines. Amines can be tethered to the cation or the anion, or dissolved in RTILs, providing a wide range of chemical solvents for CO2 capture. However, despite all of their promising features, RTILs do have drawbacks to use in CO2 separations, which have been overlooked as appropriate comparisons of RTILs to common organic solvents and polymers have not been reported. A thorough summary of the capabilitiesand limitationsof imidazolium-based RTILs in CO2-based separations with respect to a variety of materials is thus provided.
Tropical reefs have been subjected to a range of anthropogenic pressures such as global climate change, overfishing and eutrophication that have raised questions about the prominence of macroalgae on ...tropical reefs, whether they pose a threat to biodiversity, and how they may influence the function of tropical marine ecosystems.
We synthesise current understanding of the structure and function of tropical macroalgal reefs and how they may support various ecosystem goods and services. We then forecast how key stressors may alter the role of macroalgal reefs in tropical seascapes of the Anthropocene.
High levels of primary productivity from tropical canopy macroalgae, which rivals that of other key producers (e.g., corals and turf algae), can be widely dispersed across tropical seascapes to provide a boost of secondary productivity in a range of biomes that include coral reefs, and support periodic harvests of macroalgal biomass for industrial and agricultural uses. Complex macroalgal reefs that comprise a mixture of canopy and understorey taxa can also provide key habitats for a diverse community of epifauna, as well as juvenile and adult fishes that are the basis for important tropical fisheries.
Key macroalgal taxa (e.g., Sargassum) that form complex macroalgal reefs are likely to be sensitive to future climate change. Increases in maximum sea temperature, in particular, could depress biomass production and/or drive phenological shifts in canopy formation that will affect their capacity to support tropical marine ecosystems.
Macroalgal reefs can support a suite of tropical marine ecosystem functions when embedded within an interconnected mosaic of habitat types. Habitat connectivity is, therefore, essential if we are to maintain tropical marine biodiversity alongside key ecosystem goods and services. Consequently, complex macroalgal reefs should be treated as a key ecological asset in strategies for the conservation and management of diverse tropical seascapes.
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Collagen structure has been shown to influence tumor cell invasion, metastasis and clinical outcome in breast cancer. However, it remains unclear how it affects other solid cancers. Here we utilized ...multi-photon laser scanning microscopy and Second Harmonic Generation to identify alterations to collagen fiber structure within the tumor stroma of head & neck, esophageal and colorectal cancers. Image segmentation algorithms were then applied to quantitatively characterize these morphological changes, showing that elongated collagen fibers significantly correlated with poor clinical outcome (Log Rank p < 0.05). We used TGF-β treatment to model fibroblast conversion to smooth muscle actin SMA-positive cancer associated fibroblasts (CAFs) and found that these cells induce the formation of elongated collagen fibers in vivo. However, proteomic/transcriptomic analysis of SMA-positive CAFs cultured ex-vivo showed significant heterogeneity in the expression of genes with collagen fibril organizing gene ontology. Notably, stratifying patients according to stromal SMA-positivity and collagen fiber elongation was found to provide a highly significant correlation with poor survival in all 3 cancer types (Log Rank p ≤ 0.003). In summary, we show that increased collagen fiber length correlates with poor patient survival in multiple tumor types and that only a sub-set of SMA-positive CAFs can mediate the formation of this collagen structure.
Fibrillar deposits of highly phosphorylated tau are a key pathological feature of several neurodegenerative tauopathies including Alzheimer's disease (AD) and some frontotemporal dementias. ...Increasing evidence suggests that the presence of these end-stage neurofibrillary lesions do not cause neuronal loss, but rather that alterations to soluble tau proteins induce neurodegeneration. In particular, aberrant tau phosphorylation is acknowledged to be a key disease process, influencing tau structure, distribution, and function in neurons. Although typically described as a cytosolic protein that associates with microtubules and regulates axonal transport, several additional functions of tau have recently been demonstrated, including roles in DNA stabilization, and synaptic function. Most recently, studies examining the trans-synaptic spread of tau pathology in disease models have suggested a potential role for extracellular tau in cell signaling pathways intrinsic to neurodegeneration. Here we review the evidence showing that tau phosphorylation plays a key role in neurodegenerative tauopathies. We also comment on the tractability of altering phosphorylation-dependent tau functions for therapeutic intervention in AD and related disorders.
Room-temperature ionic liquids (RTILs) with polymerizable groups can be readily converted into solid, dense poly(RTILs) for use as gas separation membranes. A series of RTIL monomers with varying ...length n-alkyl substituents were synthesized and converted into polymer films. These membranes were tested for their performance in separations involving CO2, N2, and CH4. CO2 permeability was observed to increase in a nonlinear fashion as the n-alkyl substituent was lengthened. CO2/N2 separation performance was relatively unaffected as CO2 permeability increased. Plotting the performance of these membranes on a “Robeson plot” for CO2/N2 shows that first-generation poly(RTILs) “hug” the “upper bound” of the chart, indicating that they perform as well or better than many other polymers for this separation. The CO2/CH4 separation is less impressive when compared to other polymer membranes on a “Robeson plot”, but poly(RTILs) perform as well or better than molten RTILs do in bulk fluid gas absorptions for that gas pair. Furthermore, poly(RTILs) were determined to be able to absorb about twice as much CO2 as their liquid analogues, an important factor which may give them potential use as gas and vapor sorbents.
Stalk lodging (breaking of agricultural plant stalks prior to harvest) is a multi-billion dollar a year problem. Stalk lodging occurs when bending moments induced by a combination of external loading ...(e.g. wind) and self-loading (e.g. the plant's own weight) exceed the stalk bending strength of plant stems. Previous studies have investigated external loading and self-loading of plants as separate and independent phenomena. However, these two types of loading are highly interconnected and mutually dependent. The purpose of this paper is twofold: (1) to investigate the combined effect of external loads and plant weight on the flexural response of plant stems, and (2) to provide a generalized framework for accounting for self-weight during mechanical phenotyping experiments used to predict stalk lodging resistance.
A mathematical methodology for properly accounting for the interconnected relationship between self-loading and external loading of plants stems is presented. The method was compared to numerous finite element models of plants stems and found to be highly accurate. The resulting interconnected set of equations from the derivation were used to produce user-friendly applications by presenting (1) simplified self-loading correction factors for common loading configurations of plants, and (2) a generalized Microsoft Excel framework that calculates the influence of self-loading on crop stems. Results indicate that ignoring the effects of self-loading when calculating stalk flexural stiffness is appropriate for large and stiff plants such as maize, bamboo, and sorghum. However, significant errors result when ignoring the effects of self-loading in smaller plants with larger relative grain sizes, such as rice (8% error) and wheat (16% error).
Properly accounting for self-weight can be critical to determining the structural response of plant stems. Equations and tools provided herein enable researchers to properly account for the plant's weight during mechanical phenotyping experiments used to determine stalk lodging resistance.
A series of four imidazolium salts containing increasing lengths of fluoroalkyl substituents was synthesized. The three that exist as molten salts at 296
K were tested for their gas separation ...properties relating to CO
2, O
2, N
2 and CH
4 using a supported ionic liquid membrane (SILM) configuration. These fluoroalkyl-functionalized room-temperature ionic liquids (RTILs) were found to exhibit ideal selectivities for CO
2/N
2 separation that were lower than their alkyl-functionalized analogues, but higher ideal selectivity for CO
2/CH
4 separation. The differences in performance of fluoroalkyl-functionalized RTILs relative to their alkyl-functionalized counterparts are explained through the use of solubility parameters, group contributions and in context of the classically observed deviations of fluorocarbons from “regular” solution behavior.
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