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1.
Photochem Photobiol ; 2024 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-38433456

RESUMEN

Nucleic acids, lipids, and other cell components can be found within different types of extracellular vesicles (EVs), which include apoptotic bodies (ABs), large extracellular vesicles (LEVs), and small extracellular vesicles (SEVs). Release of LEVs from cells can be reduced by genetic or pharmacological inhibition of the enzyme acid sphinogomyelinase (aSMase), and indeed several studies have demonstrated a role for the clinically approved aSMase inhibitor imipramine in blocking LEV release, including in response to UVB exposure. Given that exposure of keratinocytes to UVB radiation results in the generation of UVR photoproducts in DNA that can subsequently be found in association with ABs and SEVs, we examined how imipramine impacts the release of extracellular DNA containing UVR photoproducts at an early time point after UVR exposure. Using several different model systems, including cultured keratinocytes in vitro, discarded human surgical skin ex vivo, and skin biopsies obtained from treated human subjects, these pilot studies suggest that imipramine treatment stimulates the release of CPD-containing, SEV-associated DNA. These surprising findings indicate that LEV and SEV generation pathways could be linked in UVB-irradiated cells and that imipramine may exacerbate the systemic effects of extracellular UVR-damaged DNA throughout the body.

2.
Photochem Photobiol ; 2024 Jan 29.
Artículo en Inglés | MEDLINE | ID: mdl-38287748

RESUMEN

Photosensitivity can be due to numerous causes. The photosensitivity associated with deficiency of xeroderma pigmentosum type A (XPA) has been previously shown to be associated with excess levels of the lipid mediator platelet-activating factor (PAF) generated by the keratinocyte. As PAF has been reported to trigger the production of subcellular microvesicle particles (MVP) due to the enzyme acid sphingomyelinase (aSMase), the goal of these studies was to discern if PAF and aSMase could serve as therapeutic targets for the XPA deficiency photosensitivity. HaCaT keratinocytes lacking XPA generated greater levels of MVP in comparison to control cells. Mice deficient in XPA also generated enhanced MVP levels in skin and in plasma in response to UV radiation. Use of a genetic strategy with mice deficient in both XPA and PAF receptors revealed that these mice generated less MVP release as well as decreased skin erythema and cytokine release compared to XPA knockout mice alone. Finally, the aSMase inhibitor imipramine blocked UV-induced MVP release in HaCaT keratinocytes, as well as XPA knockout mice. These studies support the concept that the photosensitivity associated with XPA involves PAF- and aSMase-mediated MVP release and provides a potential pharmacologic target in treating this form of photosensitivity.

3.
STAR Protoc ; 5(1): 102838, 2024 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-38244199

RESUMEN

UV radiation induces the formation of adducts in genomic DNA within cells that are later found to be present in cell-free fractions associated with extracellular vesicles (EVs) outside of cells. Here, we present a protocol for isolating UV photoproducts in extracellular DNA released from UVB-irradiated cells via differential centrifugation. We then detail steps for monitoring the DNA adducts using DNA immunoblotting. This protocol can be applied for detection of DNA adducts in EVs from cell culture and skin explant models. For complete details on the use and execution of this protocol, please refer to Carpenter et al.1.


Asunto(s)
Aductos de ADN , Daño del ADN , ADN/genética , Rayos Ultravioleta/efectos adversos , Piel
4.
Phys Rev Lett ; 130(24): 242501, 2023 Jun 16.
Artículo en Inglés | MEDLINE | ID: mdl-37390416

RESUMEN

Excited-state spectroscopy from the first experiment at the Facility for Rare Isotope Beams (FRIB) is reported. A 24(2)-µs isomer was observed with the FRIB Decay Station initiator (FDSi) through a cascade of 224- and 401-keV γ rays in coincidence with ^{32}Na nuclei. This is the only known microsecond isomer (1 µs≤T_{1/2}<1 ms) in the region. This nucleus is at the heart of the N=20 island of shape inversion and is at the crossroads of the spherical shell-model, deformed shell-model, and ab initio theories. It can be represented as the coupling of a proton hole and neutron particle to ^{32}Mg, ^{32}Mg+π^{-1}+ν^{+1}. This odd-odd coupling and isomer formation provides a sensitive measure of the underlying shape degrees of freedom of ^{32}Mg, where the onset of spherical-to-deformed shape inversion begins with a low-lying deformed 2^{+} state at 885 keV and a low-lying shape-coexisting 0_{2}^{+} state at 1058 keV. We suggest two possible explanations for the 625-keV isomer in ^{32}Na: a 6^{-} spherical shape isomer that decays by E2 or a 0^{+} deformed spin isomer that decays by M2. The present results and calculations are most consistent with the latter, indicating that the low-lying states are dominated by deformation.


Asunto(s)
Núcleo Celular , Corazón , Isótopos , Neutrones
5.
Data Brief ; 48: 109299, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-37383803

RESUMEN

The use of inoculants added to precursor powder is a method of influencing grain growth during fabrication. Niobium carbide (NbC) particles have been added to IN718 gas atomised powder for additive manufacturing via laser-blown-powder directed-energy-deposition (LBP-DED). The collected data in this study reveals the effects of the NbC particles on the grain structure, texture and elastic properties, and oxidative properties of LBP-DED IN718 in the As-DED and heat-treated conditions. The microstructure was investigated using X-ray diffraction (XRD), scanning electron microscopy (SEM) coupled with electron backscattered diffraction (EBSD), and transmission electron microscopy (TEM) coupled with energy dispersive X-ray spectroscopy (EDS). Resonant ultrasound spectroscopy (RUS) was used to measure the elastic properties and phase transitions during standard heat treatments. Thermogravimetric analysis (TGA) is used to probe the oxidative properties at 650°C.

6.
Artículo en Inglés | MEDLINE | ID: mdl-36266177

RESUMEN

OBJECTIVES: To determine the types of products used to treat dry mouth and their perceived effectiveness, the relationship between salivary function and xerostomia symptoms, and whether salivary function predicts response to management strategies. STUDY DESIGN: Cross-sectional study of 87 patients with dry mouth and documented unstimulated whole salivary (UWS) and stimulated whole salivary (SWS) flow rates. Participants completed a questionnaire assessing dry mouth complaints and symptoms and effectiveness of specific dry mouth products. RESULTS: Mean (SD) age was 61.7 (12.9) years including 78 (90%) females. 47 (54%) participants had Sjögren's disease. Oral dryness symptoms (0-10 scale) rated highest with a mean (SD) of 7.2 (2.17); other symptoms scored from 3.4 to 5.1. Lower levels of UWS and SWS were associated with worse dry mouth and difficulty speaking, while lower levels of SWS flow alone were associated with greater difficulty swallowing and a decline in taste. More than half of the participants (55%, n = 48) reported using ≥4 dry mouth products. Participants with normal SWS flow had significantly better responses to lozenges and prescription products. CONCLUSIONS: Patients with dry mouth and normal stimulated flow rates (i.e., residual salivary capacity) respond better to stimulatory products (parasympathomimetic and lozenges). Salivary flow rate assessments may help with recommendations of dry mouth products.


Asunto(s)
Síndrome de Sjögren , Xerostomía , Femenino , Humanos , Persona de Mediana Edad , Masculino , Satisfacción del Paciente , Estudios Transversales , Síndrome de Sjögren/complicaciones , Encuestas y Cuestionarios , Saliva
7.
Phys Rev Lett ; 129(21): 212501, 2022 Nov 18.
Artículo en Inglés | MEDLINE | ID: mdl-36461950

RESUMEN

New half-lives for exotic isotopes approaching the neutron drip-line in the vicinity of N∼28 for Z=12-15 were measured at the Facility for Rare Isotope Beams (FRIB) with the FRIB decay station initiator. The first experimental results are compared to the latest quasiparticle random phase approximation and shell-model calculations. Overall, the measured half-lives are consistent with the available theoretical descriptions and suggest a well-developed region of deformation below ^{48}Ca in the N=28 isotones. The erosion of the Z=14 subshell closure in Si is experimentally confirmed at N=28, and a reduction in the ^{38}Mg half-life is observed as compared with its isotopic neighbors, which does not seem to be predicted well based on the decay energy and deformation trends. This highlights the need for both additional data in this very exotic region, and for more advanced theoretical efforts.

8.
J Invest Dermatol ; 142(11): 3062-3070.e3, 2022 11.
Artículo en Inglés | MEDLINE | ID: mdl-35691362

RESUMEN

Solar radiation induces the formation of cyclobutane pyrimidine dimers (CPDs) and other UV photoproducts in the genomic DNA of epidermal keratinocytes. Although CPDs have been detected in urine from UV- and sun-exposed individuals, the pathway by which they arrive there and the mechanisms by which UV-induced DNA damage in the skin has systemic effects throughout the body are not clear. Consistent with previous reports that DNA associates with small extracellular vesicles that are released from a variety of cell types, we observed that a small fraction of CPDs formed in genomic DNA after UVB exposure can later be detected in the culture medium. These extracellular CPDs are found within large fragments of histone-associated DNA and are released in a time- and UVB dose‒dependent manner. Moreover, studies with both cultured cells and human skin explants revealed that CPD release into the extracellular environment is blocked by caspase inhibition, which indicates a role for apoptotic signaling in CPD release from UVB-irradiated keratinocytes. Finally, we show that this released CPD-containing DNA can be taken up by other keratinocytes. These results therefore provide possible mechanisms for the export of damaged DNA from UVB-irradiated cells and for systemic effects of UVB exposure throughout the body.


Asunto(s)
Caspasas , Dímeros de Pirimidina , Humanos , Dímeros de Pirimidina/efectos de la radiación , Caspasas/metabolismo , Histonas/metabolismo , Rayos Ultravioleta/efectos adversos , Queratinocitos/metabolismo , Daño del ADN , ADN/metabolismo
9.
JID Innov ; 1(3): 100023, 2021 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-34909723

RESUMEN

Spironolactone (SP) is used to treat a variety of disparate disease states ranging from heart failure to acne through antagonism of the mineralocorticoid and androgen receptors. Although normally taken as an oral medication, recent studies have explored the topical application of SP onto the skin. However, because SP induces the proteolytic degradation of the XPB protein, which plays critical roles in DNA repair and transcription, there may be safety concerns with the use of topical SP. In this study, we show that the topical application of a high concentration of either SP or its metabolite canrenone onto human skin ex vivo lowers XPB protein levels and induces toxic responses in the epidermis. Interestingly, although SP and canrenone both inhibit cell proliferation, induce replication stress responses, and stimulate apoptotic signaling at high concentrations in cultured keratinocytes in vitro, these effects were not correlated with XPB protein loss. Thus, high concentrations of SP and canrenone likely inhibit cell proliferation and induce toxicity through additional mechanisms to XPB proteolytic degradation. This work suggests that care may need to be taken when using high concentrations of SP directly on human skin.

10.
Phys Rev Lett ; 127(20): 202501, 2021 Nov 12.
Artículo en Inglés | MEDLINE | ID: mdl-34860042

RESUMEN

Two long-standing puzzles in the decay of ^{185}Bi, the heaviest known proton-emitting nucleus are revisited. These are the nonobservation of the 9/2^{-} state, which is the ground state of all heavier odd-A Bi isotopes, and the hindered nature of proton and α decays of its presumed 60-µs 1/2^{+} ground state. The ^{185}Bi nucleus has now been studied with the ^{95}Mo(^{93}Nb,3n) reaction in complementary experiments using the Fragment Mass Analyzer and Argonne Gas-Filled Analyzer at Argonne National Laboratory's ATLAS facility. The experiments have established the existence of two states in ^{185}Bi; the short-lived T_{1/2}=2.8_{-1.0}^{+2.3} µs, proton- and α-decaying ground state, and a 58(2)-µs γ-decaying isomer, the half-life of which was previously attributed to the ground state. The reassignment of the ground-state lifetime results in a proton-decay spectroscopic factor close to unity and represents the only known example of a ground-state proton decay to a daughter nucleus (^{184}Pb) with a major shell closure. The data also demonstrate that the ordering of low- and high-spin states in ^{185}Bi is reversed relative to the heavier odd-A Bi isotopes, with the intruder-based 1/2^{+} configuration becoming the ground, similar to the lightest At nuclides.

11.
Data Brief ; 39: 107570, 2021 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-34825034

RESUMEN

The additive manufacturing (AM) of components through laser-blown-powder directed-energy-deposition (LBP-DED) is highly applicable to the repair of aerospace components. Fabrication of superalloys with this technique, as with other AM methods, often encounters complications that include the formation of undesired phases, irregular microstructure and texture leading to anisotropic elastic properties. Heat treatments and other post-processing techniques can be used to mitigate these issues. The collected data demonstrates the effects of different heat treatment protocols on the microstructure, elastic properties, and hardness of LBP-DED IN718. In this study eight different heat treatment were used to investigate the effects of treatment time and temperature. The microstructure was investigated through SEM, with XRD and EDX used for phase analysis. The texture was characterised using SEM coupled with EBSD and the elastic properties were determined from resonant ultrasound spectroscopy.

12.
Phys Rev Lett ; 127(13): 139902, 2021 Sep 24.
Artículo en Inglés | MEDLINE | ID: mdl-34623869

RESUMEN

This corrects the article DOI: 10.1103/PhysRevLett.114.082501.

13.
Rev Sci Tech ; 40(1): 91-104, 2021 Jun.
Artículo en Inglés, Francés, Español | MEDLINE | ID: mdl-34140738

RESUMEN

The availability of rapid, highly sensitive and specific molecular and serologic diagnostic assays, such as competitive enzyme-linked immunosorbent assay (cELISA), has expedited the diagnosis of emerging transboundary animal diseases, including bluetongue (BT) and African horse sickness (AHS), and facilitated more thorough characterisation of their epidemiology. The development of assays based on real-time, reverse-transcription polymerase chain reaction (RT-PCR) to detect and identify the numerous serotypes of BT virus (BTV) and AHS virus (AHSV) has aided in-depth studies of the epidemiology of BTV infection in California and AHSV infection in South Africa. The subsequent evaluation of pan-serotype, real-time, RT-PCR-positive samples through the use of serotype-specific RT-PCR assays allows the rapid identification of virus serotypes, reducing the need for expensive and time-consuming conventional methods, such as virus isolation and serotype-specific virus neutralisation assays. These molecular assays and cELISA platforms provide tools that have enhanced epidemiologic surveillance strategies and improved our understanding of potentially altered Culicoides midge behaviour when infected with BTV. They have also supported the detection of subclinical AHSV infection of vaccinated horses in South Africa. Moreover, in conjunction with whole genome sequence analysis, these tests have clarified that the mechanism behind recent outbreaks of AHS in the AHS-controlled area of South Africa was the result of the reversion to virulence and/or genome reassortment of live attenuated vaccine viruses. This review focuses on the use of contemporary molecular diagnostic assays in the context of recent epidemiologic studies and explores their advantages over historic virus isolation and serologic techniques.


La disponibilité d'essais diagnostiques moléculaires et sérologiques rapides, hautement sensibles et spécifiques tels que l'épreuve immuno-enzymatique de compétition (ELISAc), a accéléré le diagnostic des maladies animales transfrontalières émergentes, dont la fièvre catarrhale ovine (FCO) et la peste équine, et contribué à dresser un tableau épidémiologique plus complet de ces maladies. Grâce à la mise au point d'essais basés sur l'amplification en chaîne par polymérase en temps réel couplée à une transcription inverse (RT­PCR) qui permettent de détecter et d'identifier les nombreux sérotypes du virus de la fièvre catarrhale du mouton et du virus de la peste équine, des études approfondies ont pu être conduites sur l'épidémiologie de l'infection par le virus de la fièvre catarrhale du mouton en Californie et de l'infection par le virus de la peste équine en Afrique du Sud. L'évaluation postérieure des échantillons positifs à une RT­PCR en temps réel de groupe (détectant le virus quel que soit le sérotype) au moyen de RT­PCR spécifiques de chaque sérotype permet d'identifier rapidement le sérotype causal et de limiter le recours à des méthodes classiques onéreuses et chronophages comme l'isolement viral ou les essais de neutralisation virale spécifiques de chaque sérotype. Les outils fournis par ces essais moléculaires et par les plateformes ELISAc ont renforcé les stratégies de surveillance épidémiologique et permis de mieux connaître les altérations potentielles de comportement chez les tiques Culicoides infectées par le virus de la fièvre catarrhale du mouton. Ils ont également contribué à détecter les cas d'infection asymptomatique par le virus de la peste équine chez des chevaux vaccinés en Afrique du Sud. En outre, associés avec l'analyse de séquences du génome entier, ces tests ont révélé que le mécanisme sous-jacent aux récents foyers de peste équine dans la zone de contrôle en Afrique du Sud correspondait à une réversion vers la virulence et/ou à un réassortiment du génome des souches de vaccin à virus vivant atténué. Les auteurs passent en revue l'utilisation des essais de diagnostic moléculaire de nouvelle génération dans le contexte de récentes études épidémiologiques et cherchent à établir leurs avantages par rapport aux techniques classiques d'isolement viral et de recherche sérologique.


La existencia de ensayos moleculares y serológicos de diagnóstico rápidos y de gran sensibilidad y especificidad, como el ensayo inmunoenzimático de competición (ELISAc), ha acelerado el diagnóstico de enfermedades animales transfronterizas emergentes, como la lengua azul o la peste equina, y facilitado una caracterización más exhaustiva de su epidemiología. La creación de ensayos basados en la reacción en cadena de la polimerasa acoplada a transcripción inversa (RT?PCR) en tiempo real para detectar y caracterizar los numerosos serotipos de los virus de la lengua azul y la peste equina ha ayudado a estudiar a fondo la epidemiología de sendos episodios infecciosos causados por el virus de la lengua azul en California y por el virus de la peste equina en Sudáfrica. El subsiguiente análisis de las muestras positivas a la prueba de RT?PC en tiempo real de cualquier serotipo con empleo de ensayos RT?PCR dirigidos específicamente contra uno u otro serotipo permite identificar rápidamente los serotipos víricos, lo que hace menos necesario el uso de métodos convencionales más caros y largos, como el aislamiento del virus o técnicas de neutralización vírica adaptadas específicamente a un serotipo. Estos dispositivos de ensayo molecular o de ELISAc ponen a nuestra disposición herramientas que potencian las estrategias de vigilancia epidemiológica y ayudan a conocer mejor las eventuales alteraciones del comportamiento de los jejenes Culicoides al ser infectados por el virus de la lengua azul. Estas técnicas han ayudado también a detectar en Sudáfrica casos de infección asintomática por el virus de la peste equina en caballos vacunados. Estas pruebas, además, empleadas en combinación con el análisis de secuencias genómicas completas, han servido para aclarar que el mecanismo subyacente a los recientes brotes de peste equina surgidos en la zona de Sudáfrica donde la enfermedad estaba bajo control fue fruto de la reversión a la virulencia y/o el reordenamiento genómico de virus vacunales atenuados. Los autores, centrándose en el uso de modernos ensayos moleculares de diagnóstico como parte de recientes estudios epidemiológicos, examinan las ventajas que ofrecen en comparación con las tradicionales técnicas serológicas y de aislamiento vírico.


Asunto(s)
Virus de la Enfermedad Equina Africana , Enfermedad Equina Africana , Virus de la Lengua Azul , Lengua Azul , Enfermedades de los Caballos , Enfermedades de las Ovejas , Enfermedad Equina Africana/diagnóstico , Enfermedad Equina Africana/epidemiología , Virus de la Enfermedad Equina Africana/genética , Animales , Lengua Azul/diagnóstico , Lengua Azul/epidemiología , Virus de la Lengua Azul/genética , Caballos , Ovinos , Sudáfrica/epidemiología
15.
Appl Radiat Isot ; 172: 109693, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-33774323

RESUMEN

225Ac is a valuable medical radionuclide for targeted α therapy, but 227Ac is an undesirable byproduct of an accelerator-based synthesis method under investigation. Sufficient detector sensitivity is critical for quantifying the trace impurity of 227Ac, with the 227Ac/225Ac activity ratio predicted to be approximately 0.15% by end-of-bombardment (EOB). Superconducting transition edge sensor (TES) microcalorimeters offer high resolution energy spectroscopy using the normal-to-superconducting phase transition to measure small changes in temperature. By embedding 225Ac production samples in a gold foil thermally coupled to a TES microcalorimeter we can measure the decay energies of the radionuclides embedded with high resolution and 100% detection efficiency. This technique, known as decay energy spectroscopy (DES), collapses several peaks from α decays into single Q-value peaks. In practice there are more complex factors in the interpretation of data using DES, which we will discuss herein. Using this technique we measured the EOB 227Ac impurity to be (0.142 ± 0.005)% for a single production sample. This demonstration has shown that DES is a useful tool for quantitative measurements of complicated spectra.


Asunto(s)
Actinio/química , Análisis Espectral/métodos , Calorimetría/métodos , Temperatura
16.
J Phys Condens Matter ; 33(12)2021 Apr 08.
Artículo en Inglés | MEDLINE | ID: mdl-33007773

RESUMEN

Resonant ultrasound spectroscopy has been used to characterise strain coupling and relaxation behavior associated with magnetic/magnetoelectric phase transitions in GdMnO3, TbMnO3and TbMn0.98Fe0.02O3through their influence on elastic/anelastic properties. Acoustic attenuation ahead of the paramagnetic to colinear-sinusoidal incommensurate antiferromagnetic transition at ∼41 K correlates with anomalies in dielectric properties and is interpreted in terms of Debye-like freezing processes. A loss peak at ∼150 K is related to a steep increase in electrical conductivity with a polaron mechanism. The activation energy,Ea, of ≳0.04 eV from a loss peak at ∼80 K is consistent with the existence of a well-defined temperature interval in which the paramagnetic structure is stabilised by local, dynamic correlations of electric and magnetic polarisation that couple with strain and have relaxation times in the vicinity of ∼10-6s. Comparison with previously published data for Sm0.6Y0.4MnO3confirms that this pattern may be typical for multiferroic orthorhombicRMnO3perovskites (R= Gd, Tb, Dy). A frequency-dependent loss peak near 10 K observed for TbMnO3and TbMn0.98Fe0.02O3, but not for GdMnO3, yieldedEa⩾ ∼0.002 eV and is interpreted as freezing of some magnetoelastic component of the cycloid structure. Small anomalies in elastic properties associated with the incommensurate and cycloidal magnetic transitions confirm results from thermal expansion data that the magnetic order parameters have weak but significant coupling with strain. Even at strain magnitudes of ∼0.1-1‰, polaron-like strain effects are clearly important in defining the development and evolution of magnetoelectric properties in these materials. Strains associated with the cubic-orthorhombic transition due to the combined Jahn-Teller/octahedral tilting transition in the vicinity of 1500 K are 2-3 orders of magnitude greater. It is inevitable that ferroelastic twin walls due to this transition would have significantly different magnetoelectric properties from homogeneous domains due to magnetoelastic coupling with steep strain gradients.

17.
J Phys Condens Matter ; 33(9): 095402, 2021 Mar 03.
Artículo en Inglés | MEDLINE | ID: mdl-33202391

RESUMEN

Domain walls in Cu-Cl boracite develop as a consequence of an improper ferroelastic, improper ferroelectric transition, and have attracted close interest because some are conductive and all can be mechanically written and repositioned by application of an electric field. The phase transition and its associated dynamical properties have been analysed here from the perspective of strain and elasticity. Determination of spontaneous strains from published lattice parameter data has allowed the equilibrium long-range order parameter for F [Formula: see text]3c → Pca21 to be modelled simply as being close to the order-disorder limit. High acoustic loss in the cubic phase, revealed by resonant ultrasound spectroscopy, is consistent with the presence of dynamical microdomains of the orthorhombic structure with relaxation times in the vicinity of ∼10-5-10-6 s. Low acoustic loss in the stability field of the orthorhombic structure signifies, on the other hand, that ferroelastic twin walls which develop as a consequence of the order-disorder process are immobile on this time scale. A Debye loss peak accompanied by ∼1% elastic stiffening at ∼40 K is indicative of some freezing of defects which couple with strain or of some more intrinsic freezing process. The activation energy of ⩾∼0.01-0.02 eV implies a mechanism which could involve strain relaxation clouds around local ferroelectric dipoles or freezing of polarons that determine the conductivity of twin walls.

18.
Phys Rev Lett ; 125(10): 102502, 2020 Sep 04.
Artículo en Inglés | MEDLINE | ID: mdl-32955302

RESUMEN

The low-spin structure of the semimagic ^{64}Ni nucleus has been considerably expanded: combining four experiments, several 0^{+} and 2^{+} excited states were identified below 4.5 MeV, and their properties established. The Monte Carlo shell model accounts for the results and unveils an unexpectedly complex landscape of coexisting shapes: a prolate 0^{+} excitation is located at a surprisingly high energy (3463 keV), with a collective 2^{+} state 286 keV above it, the first such observation in Ni isotopes. The evolution in excitation energy of the prolate minimum across the neutron N=40 subshell gap highlights the impact of the monopole interaction and its variation in strength with N.

19.
ACS Chem Biol ; 15(9): 2433-2443, 2020 09 18.
Artículo en Inglés | MEDLINE | ID: mdl-32786268

RESUMEN

The development and function of tissues, blood, and the immune system is dependent upon proximity for cellular recognition and communication. However, the detection of cell-to-cell contacts is limited due to a lack of reversible, quantitative probes that can function at these dynamic sites of irregular geometry. Described here is a novel chemo-genetic tool developed for fluorescent detection of protein-protein proximity and cell apposition that utilizes the Fluorogen Activating Protein (FAP) in combination with a Dye Activated by Proximal Anchoring (DAPA). The FAP-DAPA system has two protein components, the HaloTag and FAP, expressed on separate protein targets or in separate cells. The proteins function to bind and activate a compound that has the hexyl chloride (HexCl) ligand connected to malachite green (MG), the FAP fluorogen, via a poly(ethylene glycol) spacer spanning up to 28 nm. The dehalogenase protein, HaloTag, covalently binds the HexCl ligand, locally concentrating the attached MG. If the FAP is within range of the anchored fluorogen, it will bind and activate MG specifically when the bath concentration is too low to saturate the FAP receptor. A new FAP variant was isolated with a 1000-fold reduced KD of ∼10-100 nM so that the fluorogen activation reports proximity without artificially enhancing it. The system was characterized using purified FRB and FKBP fusion proteins and showed a doubling of fluorescence upon rapamycin induced complex formation. In cocultured HEK293 cells (HaloTag and FAP-expressing) fluorescence increased at contact sites across a broad range of labeling conditions, more reliably providing contact-specific fluorescence activation with the lower-affinity FAP variant. When combined with suitable targeting and expression constructs, this labeling system may offer significant improvements in on-demand detection of intercellular contacts, potentially applicable in neurological and immunological synapse measurements and other transient, dynamic biological appositions that can be perturbed using other labeling methods that stabilize these interactions.


Asunto(s)
Colorantes Fluorescentes/metabolismo , Hidrolasas/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Colorantes de Rosanilina/metabolismo , Cumarinas/química , Cumarinas/metabolismo , Fluorescencia , Colorantes Fluorescentes/química , Células HEK293 , Hexanos/química , Hexanos/metabolismo , Humanos , Hidrocarburos Clorados/química , Hidrocarburos Clorados/metabolismo , Hidrolasas/química , Ligandos , Microscopía Fluorescente , Polietilenglicoles/química , Unión Proteica , Proteínas Recombinantes de Fusión/química , Colorantes de Rosanilina/química , Serina-Treonina Quinasas TOR/metabolismo , Proteínas de Unión a Tacrolimus/metabolismo
20.
Faraday Discuss ; 223(0): 161-182, 2020 10 23.
Artículo en Inglés | MEDLINE | ID: mdl-32749434

RESUMEN

The performance of existing gas sensors often degrades in field conditions because of the loss of measurement accuracy in the presence of interferences. Thus, new sensing approaches are required with improved sensor selectivity. We are developing a new generation of gas sensors, known as multivariable sensors, that have several independent responses for multi-gas detection with a single sensor. In this study, we analyze the capabilities of natural and fabricated photonic three-dimensional (3-D) nanostructures as sensors for the detection of different gaseous species, such as vapors and non-condensable gases. We employed bare Morpho butterfly wing scales to control their gas selectivity with different illumination angles. Next, we chemically functionalized Morpho butterfly wing scales with a fluorinated silane to boost the response of these nanostructures to the vapors of interest and to suppress the response to ambient humidity. Further, we followed our previously developed design rules for sensing nanostructures and fabricated bioinspired inorganic 3-D nanostructures to achieve functionality beyond natural Morpho scales. These fabricated nanostructures have embedded catalytically active gold nanoparticles to operate at high temperatures of ≈300 °C for the detection of gases for solid oxide fuel cell (SOFC) applications. Our performance advances in the detection of multiple gaseous species with specific nanostructure designs were achieved by coupling the spectral responses of these nanostructures with machine learning (a.k.a. multivariate analysis, chemometrics) tools. Our newly acquired knowledge from studies of these natural and fabricated inorganic nanostructures coupled with machine learning data analytics allowed us to advance our design rules for sensing nanostructures toward the required gas selectivity for numerous gas monitoring scenarios at room and high temperatures for industrial, environmental, and other applications.


Asunto(s)
Técnicas Biosensibles/métodos , Gases/análisis , Aprendizaje Automático , Oro/química , Nanopartículas del Metal/química , Óptica y Fotónica , Reproducibilidad de los Resultados
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