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21.
Mark Akeson Joshua Scharff Celia M. Sharp David M. Neville Jr. 《The Journal of membrane biology》1992,125(1):81-91
Summary We used fluorescence microscopy of Madin-Darby Canine Kidney (MDCK) cells grown on polycarbonate filters to study a possible link between plasma membrane electrical potential (pm) and infectivity of vesicular stomatitis virus (VSV). Complete substitution of K+ for extracellular Na+blocks VSV infection of MDCK cells as well as baby hamster kidney (BHK) cells. When we independently perfused the apical and basal-lateral surfaces of high resistance monolayers, high K+ inhibited VSV infection of MDCK cells only when applied to the basal-lateral side; high K+ applied apically had no effect on VSV infection. This morphological specificity correlates with a large decrease in pm of MDCK cells when high K+ buffer is perfused across the basal-lateral surface. Depolarization of the plasma membrane by 130 mm basal K+ causes a sustained increase of cytosol pH in MDCK cells from 7.3 to 7.5 as reported by the fluorescent dye BCECF. Depolarization also causes a transient increase of cytosol Ca2+ from 70 to 300 nm as reported by the dye Fura-2. Neither increase could explain the block of VSV infectivity by plasma membrane depolarization. One alternative hypothesis is that pm facilitates membrane translocation of viral macromolecules as previously described for colicins, mitochondrial import proteins, and proteins secreted by Escherichia coli.We thank Kenneth Spring for many helpful discussions concerning fluorescence digitized imaging systems, James Russell for his collaboration in the design of our imaging system, Herbert Chase for suggestions on dye loading into MDCK cells, and Manfred Schubert and George Harmison for providing expertise on VSV. 相似文献
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Ana Bortolotti Ana Sánchez-Azqueta Celia M. Maya Adrián Velázquez-Campoy Juan A. Hermoso Milagros Medina Néstor Cortez 《BBA》2014
To study the role of the mobile C-terminal extension present in bacterial class of plant type NADP(H):ferredoxin reductases during catalysis, we generated a series of mutants of the Rhodobacter capsulatus enzyme (RcFPR). Deletion of the six C-terminal amino acids beyond alanine 266 was combined with the replacement A266Y, emulating the structure present in plastidic versions of this flavoenzyme. Analysis of absorbance and fluorescence spectra suggests that deletion does not modify the general geometry of FAD itself, but increases exposure of the flavin to the solvent, prevents a productive geometry of FAD:NADP(H) complex and decreases the protein thermal stability. Although the replacement A266Y partially coats the isoalloxazine from solvent and slightly restores protein stability, this single change does not allow formation of active charge-transfer complexes commonly present in the wild-type FPR, probably due to restraints of C-terminus pliability. A proton exchange process is deduced from ITC measurements during coenzyme binding. All studied RcFPR variants display higher affinity for NADP+ than wild-type, evidencing the contribution of the C-terminus in tempering a non-productive strong (rigid) interaction with the coenzyme. The decreased catalytic rate parameters confirm that the hydride transfer from NADPH to the flavin ring is considerably hampered in the mutants. Although the involvement of the C-terminal extension from bacterial FPRs in stabilizing overall folding and bent-FAD geometry has been stated, the most relevant contributions to catalysis are modulation of coenzyme entrance and affinity, promotion of the optimal geometry of an active complex and supply of a proton acceptor acting during coenzyme binding. 相似文献
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Sabrina Camargo Maik Riedl Celia Anteneodo Jürgen Kurths Thomas Penzel Niels Wessel 《PloS one》2014,9(9)
Sleep disorders are a major risk factor for cardiovascular diseases. Sleep apnea is the most common sleep disturbance and its detection relies on a polysomnography, i.e., a combination of several medical examinations performed during a monitored sleep night. In order to detect occurrences of sleep apnea without the need of combined recordings, we focus our efforts on extracting a quantifier related to the events of sleep apnea from a cardiovascular time series, namely systolic blood pressure (SBP). Physiologic time series are generally highly nonstationary and entrap the application of conventional tools that require a stationary condition. In our study, data nonstationarities are uncovered by a segmentation procedure which splits the signal into stationary patches, providing local quantities such as mean and variance of the SBP signal in each stationary patch, as well as its duration . We analysed the data of 26 apneic diagnosed individuals, divided into hypertensive and normotensive groups, and compared the results with those of a control group. From the segmentation procedure, we identified that the average duration , as well as the average variance , are correlated to the apnea-hypoapnea index (AHI), previously obtained by polysomnographic exams. Moreover, our results unveil an oscillatory pattern in apneic subjects, whose amplitude is also correlated with AHI. All these quantities allow to separate apneic individuals, with an accuracy of at least . Therefore, they provide alternative criteria to detect sleep apnea based on a single time series, the systolic blood pressure. 相似文献
26.
Julio C. Chávez José L. de la Vega-Beltrán Jessica Escoffier Pablo E. Visconti Claudia L. Trevi?o Alberto Darszon Lawrence Salkoff Celia M. Santi 《PloS one》2013,8(4)
Unlike most cells of the body which function in an ionic environment controlled within narrow limits, spermatozoa must function in a less controlled external environment. In order to better understand how sperm control their membrane potential in different ionic conditions, we measured mouse sperm membrane potentials under a variety of conditions and at different external K+ concentrations, both before and after capacitation. Experiments were undertaken using both wild-type, and mutant mouse sperm from the knock-out strain of the sperm-specific, pH-sensitive, SLO3 K+ channel. Membrane voltage data were fit to the Goldman-Hodgkin-Katz equation. Our study revealed a significant membrane permeability to both K+ and Cl− before capacitation, as well as Na+. The permeability to both K+ and Cl− has the effect of preventing large changes in membrane potential when the extracellular concentration of either ion is changed. Such a mechanism may protect against undesired shifts in membrane potential in changing ionic environments. We found that a significant portion of resting membrane potassium permeability in wild-type sperm was contributed by SLO3 K+ channels. We also found that further activation of SLO3 channels was the essential mechanism producing membrane hyperpolarization under two separate conditions, 1) elevation of external pH prior to capacitation and 2) capacitating conditions. Both conditions produced a significant membrane hyperpolarization in wild-type which was absent in SLO3 mutant sperm. Hyperpolarization in both conditions may result from activation of SLO3 channels by raising intracellular pH; however, demonstrating that SLO3-dependent hyperpolarization is achieved by an alkaline environment alone shows that SLO3 channel activation might occur independently of other events associated with capacitation. For example sperm may undergo stages of membrane hyperpolarization when reaching alkaline regions of the female genital tract. Significantly, other events associated with sperm capacitation, occur in SLO3 mutant sperm and thus proceed independently of hyperpolarization. 相似文献
27.
Microscale pigment adjustments to a tropical photosynthetically active radiation and ultraviolet (UV) environment by the intertidal turf algae Ahnfeltiopsis concinna (J. Ag.) Silva et DeCew and Laurencia mcdermidiae (J. Ag) Abbott were promoted by thalli densities that self-shade the under story portions of the same diminutive axes. Tissues of A. concinna from canopy microsites had significantly reduced levels of phycoerythrin, phycocyanin, and allophycocyanin compared to tissues from understory microsites of the same axes. Tissues of L. mcdermidiae from canopy microsites had reduced levels of only phycoerythrin compared to tissues from understory microsites. These alterations coupled with enhanced levels of carotenoid and UV-absorbing compounds in tissues from canopy compared to tissues from understory microsites indicated a pattern of remarkably sensitive photoacclimation over the ≤10-cm axes of these turf-forming rhodophytes. Microscale variation in the in vivo UV absorbance capabilities for turfs of A. concinna and L. mcdermidiae was directly related to the amount of extractable UV-absorbing compounds. An in vivo absorbance signature at ~345 nm appears to provide a method to quickly and accurately gauge the potential UV-shielding capacity of primary producers even at remarkably fine ecological scales. The capacity for highly responsive biochemical adjustments that result in marked canopy–understory distinctions coupled with a turf morphology may be crucial for macroalgal tolerance of physiological stresses associated with tropical intertidal zones. This responsive capacity allows for enhanced photoprotective mechanisms in tissues from canopy microsites while optimizing irradiance capture in deeply shaded tissues from understory microsites < 10 cm away. 相似文献
28.
Susanne Siebentritt Enrico Avancini Marcus Br Jakob Bombsch Emilie Bourgeois Stephan Buecheler Romain Carron Celia Castro Sebastien Duguay Roberto Flix Evelyn Handick Dimitrios Hariskos Ville Havu Philip Jackson Hannu‐Pekka Komsa Thomas Kunze Maria Malitckaya Roberto Menozzi Milos Nesladek Nicoleta Nicoara Martti Puska Mohit Raghuwanshi Philippe Pareige Sascha Sadewasser Giovanna Sozzi Ayodhya Nath Tiwari Shigenori Ueda Arantxa Vilalta‐Clemente Thomas Paul Weiss Florian Werner Regan G. Wilks Wolfram Witte Max Hilaire Wolter 《Liver Transplantation》2020,10(8)
29.
Susana Ruiz‐Ruiz Sergio Sanchez‐Carrillo Sergio Ciordia María C. Mena Celia Mndez‐García David Rojo Rafael Bargiela Elisa Zubeldia‐Varela Mnica Martínez‐Martínez Coral Barbas Manuel Ferrer Andrs Moya 《Aging cell》2020,19(1)
Composition of the gut microbiota changes during ageing, but questions remain about whether age is also associated with deficits in microbiome function and whether these changes occur sharply or progressively. The ability to define these deficits in populations of different ages may help determine a chronological age threshold at which deficits occur and subsequently identify innovative dietary strategies for active and healthy ageing. Here, active gut microbiota and associated metabolic functions were evaluated using shotgun proteomics in three well‐defined age groups consisting of 30 healthy volunteers, namely, ten infants, ten adults and ten elderly individuals. Samples from each volunteer at intervals of up to 6 months (n = 83 samples) were used for validation. Ageing gradually increases the diversity of gut bacteria that actively synthesize proteins, that is by 1.4‐fold from infants to elderly individuals. An analysis of functional deficits consistently identifies a relationship between tryptophan and indole metabolism and ageing (p < 2.8e?8). Indeed, the synthesis of proteins involved in tryptophan and indole production and the faecal concentrations of these metabolites are directly correlated (r2 > .987) and progressively decrease with age (r2 > .948). An age threshold for a 50% decrease is observed ca. 11–31 years old, and a greater than 90% reduction is observed from the ages of 34–54 years. Based on recent investigations linking tryptophan with abundance of indole and other “healthy” longevity molecules and on the results from this small cohort study, dietary interventions aimed at manipulating tryptophan deficits since a relatively “young” age of 34 and, particularly, in the elderly are recommended. 相似文献
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