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1.
Onderstepoort J Vet Res ; 88(1): e1-e8, 2021 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-34212734

RESUMO

Toxoplasma gondii, an obligate intracellular parasite, is the aetiological agent of toxoplasmosis, a disease that affects approximately 25% - 30% of the world's population. At present, no safe and effective vaccine exists for the prevention of toxoplasmosis. Current treatment options for toxoplasmosis are active only against tachyzoites and may also cause bone marrow toxicity. To contribute to the global search for novel agents for the treatment of toxoplasmosis, we herein report the in vitro activities of previously synthesised benzyltriazole derivatives. The effects of these compounds against T. gondii in vitro were evaluated by using a expressing green fluorescent protein (GFP) type I strain parasite (RH-GFP) and a type II cyst-forming strain of parasite (PruΔku80Δhxgprt). The frontline antitubercular drug isoniazid, designated as Frans J. Smit -isoniazid (FJS-INH), was also included in the screening as a preliminary test in view of future repurposing of this agent. Of the compounds screened, FJS-302, FJS-303, FJS-403 and FJS-INH demonstrated 80% parasite growth inhibition with IC50 values of 5.6 µg/mL, 6.8 µg/µL, 7.0 µg/mL and 19.8 µg/mL, respectively. FJS-302, FJS-303 and FJS-403 inhibited parasite invasion and replication, whereas, sulphadiazine (SFZ), the positive control, was only effective against parasite replication. In addition, SFZ induced bradyzoite differentiation in vitro, whilst FJS-302, FJS-303 and FJS-403 did not increase the bradyzoite number. These results indicate that FJS-302, FJS-303 and FJS-403 have the potential to act as a viable source of antiparasitic therapeutic agents.

2.
Am J Hum Genet ; 2021 Jun 30.
Artigo em Inglês | MEDLINE | ID: mdl-34237282

RESUMO

Multiple morphological abnormalities of the sperm flagella (MMAF)-induced asthenoteratozoospermia is a common cause of male infertility. Previous studies have identified several MMAF-associated genes, highlighting the condition's genetic heterogeneity. To further define the genetic causes underlying MMAF, we performed whole-exome sequencing in a cohort of 643 Chinese MMAF-affected men. Bi-allelic DNAH10 variants were identified in five individuals with MMAF from four unrelated families. These variants were either rare or absent in public population genome databases and were predicted to be deleterious by multiple bioinformatics tools. Morphological and ultrastructural analyses of the spermatozoa obtained from men harboring bi-allelic DNAH10 variants revealed striking flagellar defects with the absence of inner dynein arms (IDAs). DNAH10 encodes an axonemal IDA heavy chain component that is predominantly expressed in the testes. Immunostaining analysis indicated that DNAH10 localized to the entire sperm flagellum of control spermatozoa. In contrast, spermatozoa from the men harboring bi-allelic DNAH10 variants exhibited an absence or markedly reduced staining intensity of DNAH10 and other IDA components, including DNAH2 and DNAH6. Furthermore, the phenotypes were recapitulated in mouse models lacking Dnah10 or expressing a disease-associated variant, confirming the involvement of DNAH10 in human MMAF. Altogether, our findings in humans and mice demonstrate that DNAH10 is essential for sperm flagellar assembly and that deleterious bi-allelic DNAH10 variants can cause male infertility with MMAF. These findings will provide guidance for genetic counseling and insights into the diagnosis of MMAF-associated asthenoteratozoospermia.

3.
Comput Math Methods Med ; 2021: 6665357, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34194537

RESUMO

In recent years, deep learning (DNN) based methods have made leapfrogging level breakthroughs in detecting cardiac arrhythmias as the cost effectiveness of arithmetic power, and data size has broken through the tipping point. However, the inability of these methods to provide a basis for modeling decisions limits clinicians' confidence on such methods. In this paper, a Gate Recurrent Unit (GRU) and decision tree fusion model, referred to as (T-GRU), was designed to explore the problem of arrhythmia recognition and to improve the credibility of deep learning methods. The fusion model multipathway processing time-frequency domain featured the introduction of decision tree probability analysis of frequency domain features, the regularization of GRU model parameters and weight control to improve the decision tree model output weights. The MIT-BIH arrhythmia database was used for validation. Results showed that the low-frequency band features dominated the model prediction. The fusion model had an accuracy of 98.31%, sensitivity of 96.85%, specificity of 98.81%, and precision of 96.73%, indicating its high reliability and clinical significance.

4.
Nat Commun ; 12(1): 4144, 2021 07 06.
Artigo em Inglês | MEDLINE | ID: mdl-34230476

RESUMO

To investigate the duration of humoral immune response in convalescent coronavirus disease 2019 (COVID-19) patients, we conduct a 12-month longitudinal study through collecting a total of 1,782 plasma samples from 869 convalescent plasma donors in Wuhan, China and test specific antibody responses. The results show that positive rate of IgG antibody against receptor-binding domain of spike protein (RBD-IgG) to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in the COVID-19 convalescent plasma donors exceeded 70% for 12 months post diagnosis. The level of RBD-IgG decreases with time, with the titer stabilizing at 64.3% of the initial level by the 9th month. Moreover, male plasma donors produce more RBD-IgG than female, and age of the patients positively correlates with the RBD-IgG titer. A strong positive correlation between RBD-IgG and neutralizing antibody titers is also identified. These results facilitate our understanding of SARS-CoV-2-induced immune memory to promote vaccine and therapy development.


Assuntos
Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Imunoglobulina G/sangue , Receptores Virais/imunologia , SARS-CoV-2/imunologia , Adulto , Animais , Doadores de Sangue , COVID-19/terapia , Linhagem Celular , China , Chlorocebus aethiops , Convalescença , Feminino , Humanos , Imunidade Humoral/imunologia , Imunização Passiva , Memória Imunológica/imunologia , Estudos Longitudinais , Masculino , Fatores Sexuais , Glicoproteína da Espícula de Coronavírus/imunologia , Células Vero
5.
Nature ; 595(7867): 450-454, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34194039

RESUMO

Family C G-protein-coupled receptors (GPCRs) operate as obligate dimers with extracellular domains that recognize small ligands, leading to G-protein activation on the transmembrane (TM) domains of these receptors by an unknown mechanism1. Here we show structures of homodimers of the family C metabotropic glutamate receptor 2 (mGlu2) in distinct functional states and in complex with heterotrimeric Gi. Upon activation of the extracellular domain, the two transmembrane domains undergo extensive rearrangement in relative orientation to establish an asymmetric TM6-TM6 interface that promotes conformational changes in the cytoplasmic domain of one protomer. Nucleotide-bound Gi can be observed pre-coupled to inactive mGlu2, but its transition to the nucleotide-free form seems to depend on establishing the active-state TM6-TM6 interface. In contrast to family A and B GPCRs, G-protein coupling does not involve the cytoplasmic opening of TM6 but is facilitated through the coordination of intracellular loops 2 and 3, as well as a critical contribution from the C terminus of the receptor. The findings highlight the synergy of global and local conformational transitions to facilitate a new mode of G-protein activation.

6.
Z Med Phys ; 2021 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-34312047

RESUMO

INTRODUCTION: Quantitative Susceptibility Mapping (QSM) is generally acquired with full brain coverage, even though many QSM brain-iron studies focus on the deep grey matter (DGM) region only. Reducing the spatial coverage to the DGM vicinity can substantially shorten the scan time or enhance the spatial resolution without increasing scan time; however, this may lead to significant DGM susceptibility underestimation. METHOD: A recently proposed deep learning-based QSM method, namely xQSM, is investigated to assess the accuracy of dipole inversion on reduced brain coverages. The xQSM method is compared with two conventional dipole inversion methods using simulated and in vivo experiments from 4 healthy subjects at 3T. Pre-processed magnetic field maps are extended symmetrically from the centre of globus pallidus in the coronal plane to simulate QSM acquisitions of difference spatial coverages, ranging from 100% (∼32mm) to 400% (∼128mm) of the actual DGM physical size. RESULTS: The proposed xQSM network led to the lowest DGM contrast loss in both simulated and in vivo subjects, with the smallest susceptibility variation range across all spatial coverages. For the digital brain phantom simulation, xQSM improved the DGM susceptibility underestimation more than 20% in small spatial coverages, as compared to conventional methods. For the in vivo acquisition, less than 5% DGM susceptibility error was achieved in 48mm axial slabs using the xQSM network, while a minimum of 112mm coverage was required for conventional methods. It is also shown that the background field removal process performed worse in reduced brain coverages, which further deteriorated the subsequent dipole inversion. CONCLUSION: The recently proposed deep learning-based xQSM method significantly improves the accuracy of DGM QSM from small spatial coverages as compared with conventional QSM algorithms, which can shorten DGM QSM acquisition time substantially.

7.
Neuroimage ; 240: 118404, 2021 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-34280526

RESUMO

Quantitative susceptibility mapping (QSM) and R2* mapping are MRI post-processing methods that quantify tissue magnetic susceptibility and transverse relaxation rate distributions. However, QSM and R2* acquisitions are relatively slow, even with parallel imaging. Incoherent undersampling and compressed sensing reconstruction techniques have been used to accelerate traditional magnitude-based MRI acquisitions; however, most do not recover the full phase signal, as required by QSM, due to its non-convex nature. In this study, a learning-based Deep Complex Residual Network (DCRNet) is proposed to recover both the magnitude and phase images from incoherently undersampled data, enabling high acceleration of QSM and R2* acquisition. Magnitude, phase, R2*, and QSM results from DCRNet were compared with two iterative and one deep learning methods on retrospectively undersampled acquisitions from six healthy volunteers, one intracranial hemorrhage and one multiple sclerosis patients, as well as one prospectively undersampled healthy subject using a 7T scanner. Peak signal to noise ratio (PSNR), structural similarity (SSIM), root-mean-squared error (RMSE), and region-of-interest susceptibility and R2* measurements are reported for numerical comparisons. The proposed DCRNet method substantially reduced artifacts and blurring compared to the other methods and resulted in the highest PSNR, SSIM, and RMSE on the magnitude, R2*, local field, and susceptibility maps. Compared to two iterative and one deep learning methods, the DCRNet method demonstrated a 3.2% to 9.1% accuracy improvement in deep grey matter susceptibility when accelerated by a factor of four. The DCRNet also dramatically shortened the reconstruction time of single 2D brain images from 36-140 seconds using conventional approaches to only 15-70 milliseconds.

8.
Nature ; 595(7867): 455-459, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34194040

RESUMO

The calcium-sensing receptor (CaSR), a cell-surface sensor for Ca2+, is the master regulator of calcium homeostasis in humans and is the target of calcimimetic drugs for the treatment of parathyroid disorders1. CaSR is a family C G-protein-coupled receptor2 that functions as an obligate homodimer, with each protomer composed of a Ca2+-binding extracellular domain and a seven-transmembrane-helix domain (7TM) that activates heterotrimeric G proteins. Here we present cryo-electron microscopy structures of near-full-length human CaSR in inactive or active states bound to Ca2+ and various calcilytic or calcimimetic drug molecules. We show that, upon activation, the CaSR homodimer adopts an asymmetric 7TM configuration that primes one protomer for G-protein coupling. This asymmetry is stabilized by 7TM-targeting calcimimetic drugs adopting distinctly different poses in the two protomers, whereas the binding of a calcilytic drug locks CaSR 7TMs in an inactive symmetric configuration. These results provide a detailed structural framework for CaSR activation and the rational design of therapeutics targeting this receptor.

9.
Nat Commun ; 12(1): 4158, 2021 07 06.
Artigo em Inglês | MEDLINE | ID: mdl-34230497

RESUMO

Prenylated indole alkaloids featuring spirooxindole rings possess a 3R or 3S carbon stereocenter, which determines the bioactivities of these compounds. Despite the stereoselective advantages of spirooxindole biosynthesis compared with those of organic synthesis, the biocatalytic mechanism for controlling the 3R or 3S-spirooxindole formation has been elusive. Here, we report an oxygenase/semipinacolase CtdE that specifies the 3S-spirooxindole construction in the biosynthesis of 21R-citrinadin A. High-resolution X-ray crystal structures of CtdE with the substrate and cofactor, together with site-directed mutagenesis and computational studies, illustrate the catalytic mechanisms for the possible ß-face epoxidation followed by a regioselective collapse of the epoxide intermediate, which triggers semipinacol rearrangement to form the 3S-spirooxindole. Comparing CtdE with PhqK, which catalyzes the formation of the 3R-spirooxindole, we reveal an evolutionary branch of CtdE in specific 3S spirocyclization. Our study provides deeper insights into the stereoselective catalytic machinery, which is important for the biocatalysis design to synthesize spirooxindole pharmaceuticals.


Assuntos
Cicloexenos/síntese química , Cicloexenos/metabolismo , Alcaloides Indólicos/síntese química , Alcaloides Indólicos/metabolismo , Vias Biossintéticas/genética , Catálise , Técnicas de Química Sintética , Compostos de Epóxi , Fermentação , Proteínas Fúngicas/genética , Modelos Moleculares , Estrutura Molecular , Oxigenases , Penicillium/genética , Penicillium/metabolismo
10.
Sci Rep ; 11(1): 14188, 2021 Jul 09.
Artigo em Inglês | MEDLINE | ID: mdl-34244533

RESUMO

The 2019 Ridgecrest, California seismic sequence, including an Mw6.4 foreshock and Mw7.1 mainshock, represent the largest regional seismic events within the past 20 years. To obtain accurate coseismic fault-slip distribution, we used precise positioning data of small earthquakes from January 2019 to October 2020 to determine the dip parameters of the eight fault geometry, and used the Interferometric Synthetic Aperture Radar (InSAR) data processed by Xu et al. (Seismol Res Lett 91(4):1979-1985, 2020) at UCSD to constrain inversion of the fault-slip distribution of both earthquakes. The results showed that all faults were sinistral strike-slips with minor dip-slip components, exception for dextral strike-slip fault F2. Fault-slip mainly occurred at depths of 0-12 km, with a maximum slip of 3.0 m. The F1 fault contained two slip peaks located at 2 km of fault S4 and 6 km of fault S5 depth, the latter being located directly above the Mw7.1hypocenter. Two slip peaks with maximum slip of 1.5 m located 8 and 20 km from the SW endpoint of the F2 fault were also identified, and the latter corresponds to the Mw6.4 earthquake. We also analyzed the influence of different inversion parameters on the fault slip distribution, and found that the slip momentum smoothing condition was more suitable for the inversion of the earthquakes slip distribution than the stress-drop smoothing condition.

11.
Proc Natl Acad Sci U S A ; 118(28)2021 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-34264848

RESUMO

During operations, surgical mesh is commonly fixed on tissues through fasteners such as sutures and staples. Attributes of surgical mesh include biocompatibility, flexibility, strength, and permeability, but sutures and staples may cause stress concentration and tissue damage. Here, we show that the functions of surgical mesh can be significantly broadened by developing a family of materials called hydrogel-mesh composites (HMCs). The HMCs retain all the attributes of surgical mesh and add one more: adhesion to tissues. We fabricate an HMC by soaking a surgical mesh with a precursor, and upon cure, the precursor forms a polymer network of a hydrogel, in macrotopological entanglement with the fibers of the surgical mesh. In a surgery, the HMC is pressed onto a tissue, and the polymers in the hydrogel form covalent bonds with the tissue. To demonstrate the concept, we use a poly(N-isopropylacrylamide) (PNIPAAm)/chitosan hydrogel and a polyethylene terephthalate (PET) surgical mesh. In the presence a bioconjugation agent, the chitosan and the tissue form covalent bonds, and the adhesion energy reaches above 100 J⋅m-2 At body temperature, PNIPAAm becomes hydrophobic, so that the hydrogel does not swell and the adhesion is stable. Compared with sutured surgical mesh, the HMC distributes force over a large area. In vitro experiments are conducted to study the application of HMCs to wound closure, especially on tissues under high mechanical stress. The performance of HMCs on dynamic living tissues is further investigated in the surgery of a sheep.

12.
Chem Commun (Camb) ; 2021 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-34296709

RESUMO

Modular construction of polyfunctional arenes from abundant feedstocks stands as an unremitting pursue in synthetic chemistry, accelerating the discovery of drugs and materials. Herein, using the multiple C-H activation strategy with versatile imidate esters, the expedient delivery of molecular libraries of densely functionalized sulfur-containing arenes was achieved, which enabled the concise construction of biologically active molecules, such as Bipenamol.

13.
Signal Transduct Target Ther ; 6(1): 269, 2021 Jul 14.
Artigo em Inglês | MEDLINE | ID: mdl-34262014

RESUMO

Intraneuronal accumulation of hyperphosphorylated tau is a hallmark pathology shown in over twenty neurodegenerative disorders, collectively termed as tauopathies, including the most common Alzheimer's disease (AD). Therefore, selectively removing or reducing hyperphosphorylated tau is promising for therapies of AD and other tauopathies. Here, we designed and synthesized a novel DEPhosphorylation TArgeting Chimera (DEPTAC) to specifically facilitate the binding of tau to Bα-subunit-containing protein phosphatase 2A (PP2A-Bα), the most active tau phosphatase in the brain. The DEPTAC exhibited high efficiency in dephosphorylating tau at multiple AD-associated sites and preventing tau accumulation both in vitro and in vivo. Further studies revealed that DEPTAC significantly improved microtubule assembly, neurite plasticity, and hippocampus-dependent learning and memory in transgenic mice with inducible overexpression of truncated and neurotoxic human tau N368. Our data provide a strategy for selective removal of the hyperphosphorylated tau, which sheds new light for the targeted therapy of AD and related-tauopathies.

14.
Opt Express ; 29(12): 19024-19033, 2021 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-34154145

RESUMO

Enhancing photon detection efficiency and time resolution in photodetectors in the entire visible range is critical to improve the image quality of time-of-flight (TOF)-based imaging systems and fluorescence lifetime imaging (FLIM). In this work, we evaluate the gain, detection efficiency, and timing performance of avalanche photodiodes (APD) with photon trapping nanostructures for photons with 450 nm and 850 nm wavelengths. At 850 nm wavelength, our photon trapping avalanche photodiodes showed 30 times higher gain, an increase from 16% to >60% enhanced absorption efficiency, and a 50% reduction in the full width at half maximum (FWHM) pulse response time close to the breakdown voltage. At 450 nm wavelength, the external quantum efficiency increased from 54% to 82%, while the gain was enhanced more than 20-fold. Therefore, silicon APDs with photon trapping structures exhibited a dramatic increase in absorption compared to control devices. Results suggest very thin devices with fast timing properties and high absorption between the near-ultraviolet and the near infrared region can be manufactured for high-speed applications in biomedical imaging. This study paves the way towards obtaining single photon detectors with photon trapping structures with gains above 106 for the entire visible range.

15.
J Int Med Res ; 49(6): 3000605211014301, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34187217

RESUMO

OBJECTIVE: Three models were used to evaluate prostate cancer after androgen deprivation therapy (ADT) and to determine the value of detecting residual lesions after treatment. METHODS: We retrospectively analysed patients with prostate cancer who received ADT from January 2018 to June 2019. Patients were divided into ADT responder and ADT non-responder groups, and clinical risk factors were determined. Regions of interest were manually contoured on each slice on fat-saturated-T2-weighted imaging, and radiomic features were extracted. Uni- and multivariate logistic regression were used to establish radiomics, clinical and combined models. RESULTS: There were 23 ADT non-responders and 20 ADT responders. In the clinical model, total prostate-specific antigen concentration and T stage were independent predictors of efficacy (area under the curve (AUC) = 0.774). The characteristics, MinIntensity and Correlation_ angle135_offset4 indicated an effective clinical model (AUC = 0.807). GLCMEntropy_ AllDirection_offset1_SD was the best feature to differentiate residual lesions from the central gland (CG) (Lesion-CG model, AUC = 0.955). Correlation_angle135_offset4, GLCMEntropy_ AllDirection_offset4_SD and GLCMEntropy_AllDirection_offset7_SD differentiated residual lesions from the peripheral zone (PZ) (Lesion-PZ model, AUC = 0.855). The AUC for the combined model was 0.904. CONCLUSIONS: Our models can guide the clinical treatment of patients with different ADT responses. Furthermore, the radiomics model can detect prostate cancer that is non-responsive to ADT.


Assuntos
Neoplasias da Próstata , Antagonistas de Androgênios/uso terapêutico , Androgênios , Humanos , Imageamento por Ressonância Magnética , Masculino , Neoplasias da Próstata/diagnóstico por imagem , Neoplasias da Próstata/tratamento farmacológico , Estudos Retrospectivos
16.
Int J Mol Sci ; 22(9)2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-34062942

RESUMO

A field experiment was established to study sweet potato growth, starch dynamic accumulation, key enzymes and gene transcription in the sucrose-to-starch conversion and their relationships under six K2O rates using Ningzishu 1 (sensitive to low-K) and Xushu 32 (tolerant to low-K). The results indicated that K application significantly improved the biomass accumulation of plant and storage root, although treatments at high levels of K, i.e., 300-375 kg K2O ha-1, significantly decreased plant biomass and storage root yield. Compared with the no-K treatment, K application enhanced the biomass accumulation of plant and storage root by 3-47% and 13-45%, respectively, through promoting the biomass accumulation rate. Additionally, K application also enhanced the photosynthetic capacity of sweet potato. In this study, low stomatal conductance and net photosynthetic rate (Pn) accompanied with decreased intercellular CO2 concentration were observed in the no-K treatment at 35 DAT, indicating that Pn was reduced mainly due to stomatal limitation; at 55 DAT, reduced Pn in the no-K treatment was caused by non-stomatal factors. Compared with the no-K treatment, the content of sucrose, amylose and amylopectin decreased by 9-34%, 9-23% and 6-19%, respectively, but starch accumulation increased by 11-21% under K supply. The activities of sucrose synthetase (SuSy), adenosine-diphosphate-glucose pyrophosphorylase (AGPase), starch synthase (SSS) and the transcription of Susy, AGP, SSS34 and SSS67 were enhanced by K application and had positive relationships with starch accumulation. Therefore, K application promoted starch accumulation and storage root yield through regulating the activities and genes transcription of SuSy, AGPase and SSS in the sucrose-to-starch conversion.


Assuntos
Glucose-1-Fosfato Adenililtransferase/genética , Glucosiltransferases/genética , Raízes de Plantas/crescimento & desenvolvimento , Potássio/farmacologia , Amilopectina/genética , Amilose/genética , Fertilização/efeitos dos fármacos , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Ipomoea batatas/crescimento & desenvolvimento , Ipomoea batatas/metabolismo , Fotossíntese/efeitos dos fármacos , Raízes de Plantas/efeitos dos fármacos , Amido/metabolismo , Sacarose/metabolismo
17.
Elife ; 102021 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-34110282

RESUMO

Nuclear factor 90 (NF90) is a novel virus sensor that serves to initiate antiviral innate immunity by triggering stress granule (SG) formation. However, the regulation of the NF90-SG pathway remains largely unclear. We found that Tim-3, an immune checkpoint inhibitor, promotes the ubiquitination and degradation of NF90 and inhibits NF90-SG-mediated antiviral immunity. Vesicular stomatitis virus (VSV) infection induces the up-regulation and activation of Tim-3 in macrophages, which in turn recruit the E3 ubiquitin ligase TRIM47 to the zinc finger domain of NF90 and initiate a proteasome-dependent degradation via K48-linked ubiquitination at Lys297. Targeted inactivation of Tim-3 enhances the NF90 downstream SG formation by selectively increasing the phosphorylation of protein kinase R and eukaryotic translation initiation factor 2α, the expression of SG markers G3BP1 and TIA-1, and protecting mice from VSV challenge. These findings provide insights into the crosstalk between Tim-3 and other receptors in antiviral innate immunity and its related clinical significance.

18.
Vet Parasitol ; 296: 109480, 2021 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-34120030

RESUMO

Eimeria necatrix is one of the most pathogenic chicken coccidia and causes avian coccidiosis, an enteric disease of major economic importance worldwide. Eimeria parasites have complex developmental life cycles, with an exogenous phase in the environment and an endogenous phase in the chicken intestine. Oocysts excreted by chickens rapidly undergo meiosis and cell division to form eight haploid sporozoites (SZ). SZ liberated from sporocysts in the chicken intestine migrate to their preferred site of development to initiate cellular invasion. To date, almost nothing is known about the proteins that mediate parasite invasion in E. necatrix. In order to discover genes with functions involved in cellular invasion, the transcriptome profiles of E. necatrix unsporulated oocysts (UO) and SZ were analyzed using a combination of third-generation single-molecule real-time sequencing (TGS) and second-generation sequencing (SGS) followed by qRT-PCR validation. Correction of TGS long reads by SGS short reads resulted in 34,932 (UO) and 23,040 (SZ) consensus isoforms. After subsequent assembly, a total of 4949 and 4254 genes were identified from UO and SZ libraries, respectively. A total of 8376 genes were identified as differentially expressed genes (DEGs) between SZ and UO. Compared to UO, 4057 genes were upregulated and 4319 genes were downregulated in SZ. Approximately 1399 and 1758 genes were defined as stage-specific genes in SZ and UO, respectively. Gene Ontology (GO) classification and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that 2978 upregulated SZ genes were clustered into 29 GO terms, and 857 upregulated SZ genes were associated with 26 KEGG pathways. We also predicted a further 50 upregulated SZ genes and 73 upregulated UO genes encoding microneme proteins, apical membrane antigens, rhoptry neck proteins, rhoptry proteins, dense granule proteins, heat shock proteins, calcium-dependent protein kinases, cyclin-dependent kinases, cGMP-dependent protein kinase, and glycosylphosphatidylinositol-anchored surface antigens. Our data reveal new features of the E. necatrix transcriptional landscape and provide resources for the development of novel vaccine candidates against E. necatrix infection.

19.
Front Cell Infect Microbiol ; 11: 649722, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34123868

RESUMO

Herpes simplex virus type 2 (HSV2), a pathogen that causes genital herpes lesions, interferes with the host immune system via various known and unknown mechanisms. This virus has been used to study viral antigenic composition. Convalescent serum from HSV2-infected patients was used to identify viral antigens via 2-D protein electrophoresis and immunoblotting. The serum predominantly recognized several capsid scaffold proteins encoded by gene UL26.5, mainly ICP35. This protein has been primarily reported to function temporarily in viral assembly but is not expressed in mature virus particles. Further immunological studies suggested that this protein elicits specific antibody and cytotoxic T lymphocyte (CTL) responses in mice, but these responses do not result in a clinical protective effect in response to HSV2 challenge. The data suggested that immunodominance of ICP35 might be used to design an integrated antigen with other viral glycoproteins.


Assuntos
Capsídeo , Herpesvirus Humano 1 , Animais , Proteínas do Capsídeo , Herpesvirus Humano 2 , Humanos , Camundongos , Proteínas Virais
20.
Environ Pollut ; 287: 117540, 2021 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-34147784

RESUMO

Cigarette smoke extract (CSE), a complex mixture of compounds, contributes to a range of eye diseases; however, the underlying pathophysiological responses to tobacco smoke remain ambiguous. The purpose of the present study was to evaluate the cigarette smoke-induced phenotypic and transcriptomic changes in the corneal epithelium with a view to elucidating the likely underlying mechanism. Accordingly, for the first time, we characterized the genome-wide effects of CSE on the corneal epithelium. The ocular surface of the mice in the experimental groups was exposed to CSE for 1 h per day for a period of one week, while mice in the control group were exposed to preservative-free artificial tears. Corneal fluorescein staining, in vivo confocal microscopy and scanning electron microscopy were performed to examine the corneal ultrastructure. Transcriptome sequencing and bioinformatics analysis were performed followed by RT-qPCR to validate gene expression changes. The results indicate that CSE exposure disrupted the structural integrity of the superficial epithelium, decreased the density of microvilli, and compromised the corneal epithelial barrier intactness. RNA-seq revealed 667 differentially expressed genes, and functional analysis highlighted the enhancement of several biological processes such as antioxidant activity and the response to oxidative stress. Moreover, the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that glutathione metabolism and drug metabolism cytochrome P450 were the most relevant pathways contributing to the effects of CSE on the corneal epithelium. Protein-protein interaction (PPI) network analysis illustrated that GCLC, NQO1, and HMOX1 were the most relevant nodes. In conclusion, the present study indicates that CSE exposure induces changes in the phenotype and genotype of the corneal epithelium. The antioxidant response element is essential for counteracting the effects of cigarette smoke on this tissue layer. These results shed novel insights into how cigarette smoke damages this ocular surface.

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