Group 3 innate lymphoid cells (ILC3s) have demonstrated roles in promoting antibacterial immunity, maintaining epithelial barrier function, and supporting tissue repair. ILC3 alterations are ...associated with chronic inflammation and inflammatory disease; however, the characteristics and relevant regulatory mechanisms of this cell population in HIV-1 infection are poorly understood due in part to a lack of a robust model. Here, we determined that functional human ILC3s develop in lymphoid organs of humanized mice and that persistent HIV-1 infection in this model depletes ILC3s, as observed in chronic HIV-1-infected patients. In HIV-1-infected mice, effective antiretroviral therapy reversed the loss of ILC3s. HIV-1-dependent reduction of ILC3s required plasmacytoid dendritic cells (pDCs), IFN-I, and the CD95/FasL pathway, as targeted depletion or blockade of these prevented HIV-1-induced ILC3 depletion in vivo and in vitro, respectively. Finally, we determined that HIV-1 infection induces CD95 expression on ILC3s via a pDC- and IFN-I-dependent mechanism that sensitizes ILC3s to undergo CD95/FasL-mediated apoptosis. We conclude that chronic HIV-1 infection depletes ILC3s through pDC activation, induction of IFN-I, and CD95-mediated apoptosis.
Abstract
Nuclear organization has an important role in determining genome function; however, it is not clear how spatiotemporal organization of the genome relates to functionality. To elucidate this ...relationship, a method for tracking any locus of interest is desirable. Recently clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) or transcription activator-like effectors were adapted for imaging endogenous loci; however, they are mostly limited to visualization of repetitive regions. Here, we report an efficient and scalable method named SHACKTeR (Short Homology and CRISPR/Cas9-mediated Knock-in of a TetO Repeat) for live cell imaging of specific chromosomal regions without the need for a pre-existing repetitive sequence. SHACKTeR requires only two modifications to the genome: CRISPR/Cas9-mediated knock-in of an optimized TetO repeat and its visualization by TetR-EGFP expression. Our simplified knock-in protocol, utilizing short homology arms integrated by polymerase chain reaction, was successful at labeling 10 different loci in HCT116 cells. We also showed the feasibility of knock-in into lamina-associated, heterochromatin regions, demonstrating that these regions prefer non-homologous end joining for knock-in. Using SHACKTeR, we were able to observe DNA replication at a specific locus by long-term live cell imaging. We anticipate the general applicability and scalability of our method will enhance causative analyses between gene function and compartmentalization in a high-throughput manner.
Employing different bands of radar to detect precipitation information in identical regions enables the acquisition of a more comprehensive precipitation cloud structure, thereby refining the ...continuity and completeness of precipitation measurements. This study first compared the coincident data from CloudSat W-band cloud profiling radar (CPR) and Global Precipitation Measurement Mission (GPM) Ka-band precipitation radar (KaPR) from 2014 to 2017, and then a synthesis of the radar reflectivity from CPR and KaPR was attempted to obtain a complete cloud and precipitation structure. The findings of the reflectivity comparisons indicated that the echo-top height identified by CPR is on average 3.6 to 4.2 km higher than that from KaPR, due to the higher sensitivity. Because of strong attenuation of CPR by liquid-phase particles, the reflectivity below the height of the melting layer usually shows an opposite gradient to KaPR with decreasing altitude. The difference in the near-surface rain rates of CPR and KaPR was found to be related to reflectivity gradients in the vertical direction, and the larger the reflectivity gradients, the greater the differences in near-surface rain rates. For better representing the complete vertical structure of precipitation clouds and improving the consistency of the reflectivity and precipitation rate, the radar reflectivity was weighted, synthesized from CPR and KaPR based on the gradient difference of the reflectivity from the two radars. We presented the synthesis results for a stratiform cloud and a deep convective case, and Spearman’s rank correlation coefficient (rs) between the GPM combined radiometer precipitation rate and the radar reflectivity was utilized to analyze the performance of the synthesis. The consistency between synthesized reflectivity and precipitation rate in the non-liquid phase was improved compared with KaPR, and the rs of the ice and mixed phases was increased by about 12% and 10%, respectively.
Due to the rapid development of urbanization, land-use types have changed greatly, which has led to many ecological problems. Therefore, the current research objective is to solve the problems in ...existence in Jinan, so as to determine the existing landscape ecological risks and optimize the landscape structure. Using 2 m high-resolution remote sensing images and related natural economic data, this study evaluated the landscape ecological risk and constructed a full-factor ecological network in Jinan with a landscape ecological risk assessment method (ERI) and a minimum cumulative resistance model (MCR) based on landscape ecology theory. The results showed that: (1) The ERI in Jinan presented a spatial concentration of high value areas in the central and central–eastern regions, while other levels in ERI areas presented a spatial distribution around the ecological regions with high risk. (2) The important corridors were mainly distributed in the south of Jinan, which were stable and not easily destroyed. The corridors in other areas were secondary, mainly passing through cultivated land and urban greenways, which were unstable and susceptible to interference.
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•Piezo1 mechanosensitive protein could be activated by ultrasonic stimulation.•The factors in the Ca2+/CaM/MLCK pathway could be influenced by ultrasonic stimulation and the ...expression of Piezo1 protein.•Ultrasonic stimulation could modulate the permeability of the HUVECs membrane by modulating the expression of Piezo1 protein.
The acoustic radiation forces produced by ultrasonic stimulation induce shear stress on objects in the acoustic field. Piezo1, a mechanosensitive ion channel protein that is expressed on the plasma membranes of vertebrate cells, can sense shear stress and transduce it into downstream signaling. In this study, we examined the sensitivity of Piezo1 to ultrasonic stimulation and assessed its downstream biological functions in human umbilical vein endothelial cells (HUVECs). Ultrasonic stimulation using a stimulation power of 0.2 W and a frequency of 1 MHz for 10 s did not induce cell damage. However, ultrasonic stimulation induced an influx of calcium ions, which increased with an increase in the stimulation duration. Knockdown of Piezo1 protein decreased the influx of calcium ions during ultrasonic stimulation, which demonstrated that Piezo1 may be activated by the shear stress produced by ultrasonic stimulation. The influx of calcium ions in response to ultrasonic stimulation could be modulated by the Piezo1 protein level. Additionally, ultrasonic stimulation reduced the levels of downstream factors such as MLCK and ATP, which are involved in the Ca2+/CaM/MLCK pathway, by suppressing Piezo1. As the Ca2+/CaM/MLCK pathway influences the permeability of the cell membrane, the internalization of FITC-Dextran into cells under ultrasonic stimulation was validated. Ultrasonic stimulation was demonstrated to promote the increase in cell permeability, and the suppression of Piezo1 was shown to induce the decrease in cell permeability. Therefore, this study shows that ultrasonic stimulation may modulate the permeability of the membrane of HUVECs by modulating the expression of Piezo1 protein.
The three-dimensional (3D) topological insulator (TI) Bi2Se3 exhibits topologically protected, linearly dispersing Dirac surface states (SSs). To access the intriguing properties of these SSs, it is ...important to distinguish them from the coexisting two-dimensional electron gas (2DEG) on the surface. Here, we use circularly polarized light to induce the inverse spin Hall effect in a Bi2Se3 thin film at different temperatures (i.e., from 77 to 300 K). It is demonstrated that the photoinduced inverse spin Hall effect (PISHE) of the top SSs and the 2DEG can be separated based on their opposite signs. The temperature and power dependence of the PISHE also confirms our method. Furthermore, it is found that the PISHE in the 2DEG is dominated by the extrinsic mechanism, as revealed by the temperature dependence of the PISHE.
Hempseed is a nutrient-rich natural resource, and high levels of hempseed oil accumulate within hemp seeds, consisting primarily of different triglycerides. Members of the diacylglycerol ...acyltransferase (DGAT) enzyme family play critical roles in catalyzing triacylglycerol biosynthesis in plants, often governing the rate-limiting step in this process. As such, this study was designed to characterize the
(
) gene family in detail. Genomic analyses of the
revealed 10 candidate
genes that were classified into four families (
,
,
,
) based on the features of different isoforms. Members of the
family were found to be associated with large numbers of cis-acting promoter elements, including plant response elements, plant hormone response elements, light response elements, and stress response elements, suggesting roles for these genes in key processes such as development, environmental adaptation, and abiotic stress responses. Profiling of these genes in various tissues and varieties revealed varying spatial patterns of
expression dynamics and differences in expression among
varieties, suggesting that the members of this gene family likely play distinct functional regulatory functions
genes were upregulated in response to cold stress, and significant differences in the mode of regulation were observed when comparing roots and leaves, indicating that
genes may play positive roles as regulators of cold responses in hemp while also playing distinct roles in shaping the responses of different parts of hemp seedlings to cold exposure. These data provide a robust basis for further functional studies of this gene family, supporting future efforts to screen the significance of
candidate genes to validate their functions to improve hempseed oil composition.
One-unit-cell FeSe films on SrTiO
substrates are of great interest owing to significantly enlarged pairing gaps characterized by two coherence peaks at ±10 meV and ±20 meV. In-situ transport ...measurement is desired to reveal novel properties. Here, we performed in-situ microscale electrical transport and combined scanning tunneling microscopy measurements on continuous one-unit-cell FeSe films with twin boundaries. We observed two spatially coexisting superconducting phases in domains and on boundaries, characterized by distinct superconducting gaps (
~15 meV vs.
~10 meV) and pairing temperatures (T
~52.0 K vs. T
~37.3 K), and correspondingly two-step nonlinear
behavior but a concurrent Berezinskii-Kosterlitz-Thouless (BKT)-like transition occurring at
~28.7 K. Moreover, the onset transition temperature
~54 K and zero-resistivity temperature
~31 K are consistent with T
and
, respectively. Our results indicate the broadened superconducting transition in FeSe/SrTiO
is related to intrinsic electronic inhomogeneity due to distinct two-gap features and phase fluctuations of two-dimensional superconductivity.
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