Phospholipid scrambling (PLS) is a ubiquitous cellular mechanism involving the regulated bidirectional transport of phospholipids down their concentration gradient between membrane leaflets. ...ANO6/TMEM16F has been shown to be essential for Ca(2+)-dependent PLS, but controversy surrounds whether ANO6 is a phospholipid scramblase or an ion channel like other ANO/TMEM16 family members. Combining patch clamp recording with measurement of PLS, we show that ANO6 elicits robust Ca(2+)-dependent PLS coinciding with ionic currents that are explained by ionic leak during phospholipid translocation. By analyzing ANO1-ANO6 chimeric proteins, we identify a domain in ANO6 necessary for PLS and sufficient to confer this function on ANO1, which normally does not scramble. Homology modeling shows that the scramblase domain forms an unusual hydrophilic cleft that faces the lipid bilayer and may function to facilitate translocation of phospholipid between membrane leaflets. These findings provide a mechanistic framework for understanding PLS and how ANO6 functions in this process.
Quercetin is a bioactive compound that is widely used in botanical medicine and traditional Chinese medicine due to its potent antioxidant activity. In recent years, antioxidant activities of ...quercetin have been studied extensively, including its effects on glutathione (GSH), enzymatic activity, signal transduction pathways, and reactive oxygen species (ROS) caused by environmental and toxicological factors. Chemical studies on quercetin have mainly focused on the antioxidant activity of its metal ion complexes and complex ions. In this review, we highlight the recent advances in the antioxidant activities, chemical research, and medicinal application of quercetin.
Mutations in ANO5 (TMEM16E) cause limb-girdle muscular dystrophy R12. Defective plasma membrane repair is a likely mechanism. Using myofibers from Ano5 knockout mice, we show that trafficking of ...several annexin proteins, which together form a cap at the site of injury, is altered upon loss of ANO5. Annexin A2 accumulates at the wound to nearly twice the level observed in WT fibers, while annexin A6 accumulation is substantially inhibited in the absence of ANO5. Appearance of annexins A1 and A5 at the cap is likewise diminished in the Ano5 knockout. These changes are correlated with an alteration in annexin repair cap fine structure and shedding of annexin-positive vesicles. We conclude that loss of annexin coordination during repair is disrupted in Ano5 knockout mice and underlies the defective repair phenotype. Although ANO5 is a phospholipid scramblase, abnormal repair is rescued by overexpression of a scramblase-defective ANO5 mutant, suggesting a novel, scramblase-independent role of ANO5 in repair.
Microporous organic networks (MONs) that exhibit good stability and hydrophobicity are promising candidates for performing HPLC separation of small organic compounds. However, their applications in ...separating large analytes as well as biomolecules are still limited by the microporous nature of MONs. Herein, we demonstrated the fabrication of a MON‐functionalized silica (MON@SiO2), exhibiting micro and mesopores for the HPLC separations of small drugs as well as large analytes, such as flavones, nonsteroidal anti‐inflammatory drugs (NSAIDs), endocrine disrupting chemicals (EDCs), and proteins. MON was successfully modified on SiO2 microspheres to yield the uniform and mono‐dispersed MON@SiO2. The separation mechanisms and performance of the MON@SiO2 packed column were evaluated for a wide range of analytes, including neutral, acidic, basic compounds, drugs, and proteins. Compared with commercial C18 and SiO2–NH2 packed columns, the proposed MON@SiO2 column afforded superior performance in the separations of flavones, NSAIDs, EDCs, and proteins. Moreover, the MON@SiO2 column also offered good repeatability with intraday RSDs (n = 7) of <0.1%, <2.0%, <2.3%, and <0.7% for the retention time, peak height, peak area, and half peak width, respectively, for separating EDCs. This work proved the potential of using MONs in the HPLC separations of drugs and proteins.
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•Zwitterionic MMON-SO3H-NH2 is prepared for MSPE of FQs.•MMON-SO3H-NH2 offers wide linear range, low LODs and large EFs for FQs.•Ion-pair, hydrogen-bonding and π-π interactions ...dominate the extraction.•Determination of FQs in meat samples is realized.
A zwitterionic magnetic microporous organic network (MMON-SO3H-NH2) with numerous amino and sulfonic acid ion-pare binding sites was designed and synthesized for efficient magnetic solid-phase extraction (MSPE) of fluoroquinolones (FQs) from meat samples. The core-shell MMON-SO3H-NH2 offered large specific surface area, rapid magnetic responsiveness, good stability, and multiple binding sites for FQs. The density functional theory and independent gradient model evaluations confirmed hydrogen bonding, π-π and ion-pair interactions between MMON-SO3H-NH2 and FQs. Under the optimal conditions, the established MMON-SO3H-NH2-MSPE-HPLC-UV method gave wide linear range (0.15–1000 μg L−1), low limits of detection (0.05–4.5 μg L−1) and limits of quantitation (0.15–13 μg L−1), and high enrichment factors (82.1–99.6) using 3 mg of adsorbent. This work demonstrates that the preparation of zwitterionic MONs is an efficient way to promote the extraction performance of MONs for zwitterionic targets and provides an effective sample pretreatment method for enriching and monitoring FQs in complex food matrices.
Limb-girdle muscular dystrophy type 2L (LGMD2L) is a myopathy arising from mutations in
; however, information about the contribution of ANO5 to muscle physiology is lacking. To explain the role of ...ANO5 in LGMD2L, we previously hypothesized that ANO5-mediated phospholipid scrambling facilitates cell-cell fusion of mononucleated muscle progenitor cells (MPCs), which is required for muscle repair. Here, we show that heterologous overexpression of ANO5 confers Ca
-dependent phospholipid scrambling to HEK-293 cells and that scrambling is associated with the simultaneous development of a nonselective ionic current. MPCs isolated from adult
mice exhibit defective cell fusion in culture and produce muscle fibers with significantly fewer nuclei compared with controls. This defective fusion is associated with a decrease of Ca
-dependent phosphatidylserine exposure on the surface of
MPCs and a decrease in the amplitude of Ca
-dependent outwardly rectifying ionic currents. Viral introduction of
in
MPCs restores MPC fusion competence, ANO5-dependent phospholipid scrambling, and Ca
-dependent outwardly rectifying ionic currents.
-rescued MPCs produce myotubes having numbers of nuclei similar to wild-type controls. These data suggest that ANO5-mediated phospholipid scrambling or ionic currents play an important role in muscle repair.
Aberrant neurogenesis is a major factor in psychiatric and neurological disorders that have significantly attracted the attention of neuroscientists. Curcumin is a primary constituent of curcuminoid ...that exerts several positive pharmacological effects on aberrant neurogenesis. First, it is important to understand the different processes of neurogenesis, and whether their dysfunction promotes etiology as well as the development of many psychiatric and neurological disorders; then investigate mechanisms by which curcumin affects neurogenesis as an active participant in pathophysiological events. Based on scientometric studies and additional extensive research, we explore the mechanisms by which curcumin regulates adult neurogenesis and in turn affects psychiatric diseases, i.e., depression and neurological disorders among them traumatic brain injury (TBI), stroke, Alzheimer's disease (AD), Gulf War Illness (GWI) and Fragile X syndrome (FXS). This review aims to elucidate the therapeutic effects and mechanisms of curcumin on adult neurogenesis in various psychiatric and neurological disorders. Specifically, we discuss the regulatory role of curcumin in different activities of neural stem cells (NSCs), including proliferation, differentiation, and migration of NSCs. This is geared toward providing novel application prospects of curcumin in treating psychiatric and neurological disorders by regulating adult neurogenesis.
Abstract We investigate the suppression of star formation in galaxy pairs based on the isolated galaxy pair sample derived from the Sloan Digital Sky Survey survey. By comparing the star formation ...rate between late-type galaxies (LTGs) in galaxy pairs and those in the isolated environment, we detect a signal of the suppression of star formation in galaxy pairs at d p < 100 kpc and 200 kpc < d p < 350 kpc. The occurrence of the suppression of star formation in these LTGs requires their companion galaxies to have an early-type morphology ( n s > 2.5). The suppression of star formation in wide galaxy pairs at 200 kpc < d p < 350 kpc mainly occurs in massive LTGs, while in close galaxy pairs at d p < 100 kpc, it only appears in LTGs with a massive companion ( log M ⋆ > 11.0 ), nearly independent of their own stellar mass. Based on these findings, we infer that the suppression of star formation in wide galaxy pairs is actually a result of galaxy conformity, while in close galaxy pairs, it stems from the influence of hot circumgalactic medium surrounding companion galaxies.
Defects on product surfaces affect quality of the product. Machine vision provides an efficient tool for the surface defect detection. Threshold is commonly used to separate objects from the image ...background in the vision-based inspection method. The Otsu method is one of the most used approaches to decide the threshold for a satisfied result when the image histogram is bimodal, but it fails when the histogram of an image is unimodal or close to unimodal. Defects in product surfaces can range from small to large sizes, which results in distributions of the image histogram change from unimodal to bimodal. An improved Otsu method, named the weighted object variance (WOV), is proposed in this research to detect defects on product surfaces. A parameter that equals the cumulative probability of defects occurrence is weighted on the object variance of between-class variance. The weight ensures that the threshold always be a value that locates at the valley of two peaks or at the left bottom rim of a single peak histogram. It is essential to have a high detection rate and low false alarm rate for the defect detection. Experimental results demonstrate the effectiveness of the improved Otsu method in the defect detection of various surfaces. Compared to other thresholding methods such as maximum entropy, Otsu, valley-emphasis, and modified valley-emphasis methods, the WOV method provides better segmentation results.
Ca2+ -activated Cl channels play pivotal roles in the cardiovascular system. They regulate vascular smooth muscle tone and participate in cardiac action potential repolarization in some species. Ca2+ ...-activated Cl channels were recently discovered to be encoded by members of the anoctamin (Ano, also called Tmem16) superfamily, but the mechanisms of Ano1 gating by Ca2+ remain enigmatic.
The objective was to identify regions of Ano1 involved in channel gating by Ca2+.
The Ca2+ sensitivity of Ano1 was estimated from rates of current activation, and deactivation in excised patches rapidly switched between zero and high Ca2+ on the cytoplasmic side. Mutation of glutamates E702 and E705 dramatically altered Ca2+ sensitivity. E702 and E705 are predicted to be in an extracellular loop, but antigenic epitopes introduced into this loop are not accessible to extracellular antibodies, suggesting this loop is intracellular. Cytoplasmically applied membrane-impermeant sulfhydryl reagents alter the Ca2+ sensitivity of Ano1 E702C and E705C as expected if E702 and E705 are intracellular. Substituted cysteine accessibility mutagenesis of the putative re-entrant loop suggests that E702 and E705 are located adjacent to the Cl conduction pathway.
We propose an alternative model of Ano1 topology based on mutagenesis, epitope accessibility, and cysteine-scanning accessibility. These data contradict the popular re-entrant loop model by showing that the putative fourth extracellular loop (ECL 4) is intracellular and may contain a Ca2+ binding site. These studies provide new perspectives on regulation of Ano1 by Ca2+.
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