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14-3-3 proteins play important roles in plant metabolism and stress response. Tomato 14-3-3 proteins, SlTFT4 and SlTFT7, serve as hubs of plant immunity and are targeted by some pathogen effectors. Ralstonia solanacearum with more than 70 type â ¢ effectors (T3Es) is one of the most destructive plant pathogens. However, little is known on whether R. solanacearum T3Es target SlTFT4 and SlTFT7 and hence interfere with plant immunity. We first detected the associations of SlTFT4/SlTFT7 with R. solanacearum T3Es by luciferase complementation assay, and then confirmed the interactions by yeast two-hybrid approach. We demonstrated that 22 Ralstonia T3Es were associated with both SlTFT4 and SlTFT7, and five among them suppressed the hypersensitive response induced by MAPKKKα, a protein kinase which associated with SlTFT4/SlTFT7. We further demonstrated that suppression of MAPKKKα-induced HR and plant basal defense by the T3E RipAC depend on its association with 14-3-3 proteins. Our findings firstly demonstrate that R. solanacearum T3Es can manipulate plant immunity by targeting 14-3-3 proteins, SlTFT4 and SlTFT7, providing new insights into plant-R. solanacearum interactions.
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Proteínas 14-3-3 , Ralstonia solanacearum , Proteínas 14-3-3/metabolismo , Proteínas de Bactérias/metabolismo , Imunidade Vegetal , Ralstonia solanacearum/fisiologia , Doenças das Plantas , Proteínas de Plantas/metabolismoRESUMO
BACKGROUND: Elizabethkingia spp. are emerging as nosocomial pathogens causing various infections. These pathogens express resistance to a broad range of antibiotics, thus requiring antimicrobial combinations for coverage. However, possible antagonistic interactions between antibiotics have not been thoroughly explored. This study aimed to evaluate the effectiveness of antimicrobial combinations against Elizabethkingia infections, focusing on their impact on pathogenicity, including biofilm production and cell adhesion. METHODS: Double-disc diffusion, time-kill, and chequerboard assays were used for evaluating the combination effects of antibiotics against Elizabethkingia spp. We further examined the antagonistic effects of antibiotic combinations on biofilm formation and adherence to A549 human respiratory epithelial cells. Further validation of the antibiotic interactions and their implications was performed using ex vivo hamster precision-cut lung sections (PCLSs) to mimic in vivo conditions. RESULTS: Antagonistic effects were observed between cefoxitin, imipenem and amoxicillin/clavulanic acid in combination with vancomycin. The antagonism of imipenem toward vancomycin was specific to its effects on the genus Elizabethkingia. Imipenem further hampered the bactericidal effect of vancomycin and impaired its inhibition of biofilm formation and the adhesion of Elizabethkingia meningoseptica ATCC 13253 to human cells. In the ex vivo PCLS model, vancomycin exhibited dose-dependent bactericidal effects; however, the addition of imipenem also reduced the effect of vancomycin. CONCLUSIONS: Imipenem reduced the bactericidal efficacy of vancomycin against Elizabethkingia spp. and compromised its capacity to inhibit biofilm formation, thereby enhancing bacterial adhesion. Clinicians should be aware of the potential issues with the use of these antibiotic combinations when treating Elizabethkingia infections.
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Antibacterianos , Biofilmes , Infecções por Flavobacteriaceae , Imipenem , Testes de Sensibilidade Microbiana , Vancomicina , Animais , Imipenem/farmacologia , Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Humanos , Vancomicina/farmacologia , Infecções por Flavobacteriaceae/microbiologia , Infecções por Flavobacteriaceae/tratamento farmacológico , Flavobacteriaceae/efeitos dos fármacos , Aderência Bacteriana/efeitos dos fármacos , Células A549 , Cricetinae , Interações Medicamentosas , Pulmão/microbiologiaRESUMO
Actin filaments form unique structures with robust actin bundles and cytoskeletal networks affixed to the extracellular matrix and interact with neighboring cells, which are crucial structures for cancer cells to acquire a motile phenotype. This study aims to investigate a novel antitumor mechanism by which Tanshinone IIA (Tan IIA) modulates the morphology and migration of liver cancer cells via actin cytoskeleton regulation. 97H and Huh7 exhibited numerous tentacle-like protrusions that interacted with neighboring cells. Following treatment with Tan IIA, 97H and Huh7 showed a complete absence of cytoplasmic protrusion and adherens junctions, thereby effectively impeding their migration capability. The fluorescence staining of F-actin and microtubules indicated that these tentacle-like protrusions and cell-cell networks were actin-based structures that led to morphological changes after Tan IIA treatment by retracting and reorganizing beneath the membrane. Tan IIA can reverse the actin depolymerization and cell morphology alterations induced by latrunculin A. Tan IIA down-regulated actin and Rho GTPases expression significantly, as opposed to inducing Rho signaling activation. Preventing the activity of proteasomes and lysosomes had no discernible impact on the modifications in cellular structure and protein expression induced by Tan IIA. However, as demonstrated by the puromycin labeling technique, the newly synthesized proteins were significantly inhibited by Tan IIA. In conclusion, Tan IIA can induce dramatic actin cytoskeleton remodeling by inhibiting the protein synthesis of actin and Rho GTPases, resulting in the suppression of tumor growth and migration. Targeting the actin cytoskeleton of Tan IIA is a promising strategy for HCC treatment.
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Abietanos , Carcinoma Hepatocelular , Neoplasias Hepáticas , Humanos , Actinas , Proteínas rho de Ligação ao GTP/farmacologia , Proliferação de Células , Carcinoma Hepatocelular/tratamento farmacológico , Citoesqueleto , Citoesqueleto de Actina , Linhagem Celular Tumoral , ApoptoseRESUMO
Simple sequence repeats (SSRs) have been widely used for parentage testing, marker-assisted selection, and evolutionary studies. The insufficient availability of SSR markers in Bactrian camels partially accounts for the lack of systematic breeding. Therefore, we aimed to establish a comprehensive SSR dataset for the Bactrian camel. Our approach involved genome searching to locate every SSR in the genome, SSR-enriched sequencing to acquire polymorphism information, and literature research to collect published data. The resulting dataset contains 213,711 SSRs and details their characteristics, including genome coordinates, motifs, lengths, annotations, PCR primers, and polymorphism information. The dataset reveals a biased distribution of SSRs in the Bactrian camel genome, reflecting the mutation mechanism and complex evolution of SSRs. In practice, we successfully demonstrated the utility of the dataset through parentage testing using 15 randomly selected SSRs. This comprehensive dataset can facilitate systematic breeding and enable QTL mapping and GWAS of the Bactrian camel.
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Camelus , Genoma de Planta , Animais , Camelus/genética , Marcadores Genéticos , Polimorfismo Genético , Repetições de MicrossatélitesRESUMO
The mature synthetic methodologies enable us to rationally design and produce chiral nanographenes (NGs), most of which consist of multiple helical motifs. However, inherent chirality originating from twisted geometry has just emerged to be employed in chiral NGs. Herein, we report a red-emissive chiral NG constituted of orthogonally arranged two-fold twisted π-skeletons at a contorted pyrene core which contributes to optical transitions of S0âS1 and vice versa. The thus-obtained NG exhibited a robustness on its redox properties through 2e- uptake/release. The chemical oxidation generated stable radical cation whose absorption covers near-infrared I and II regions. Overall, the contorted pyrene core governs electronic nature of the chiral NG. The twist operation on NGs would be, therefore, a design strategy to alter conventional chirality induction on NGs.
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It is still very challenging to obtain colorful and long-afterglow room-temperature phosphorescent (RTP) materials from pure organic polymers. Herein, it is found that chitosan (CS), a natural polymer, not only has its own RTP, but also reacts with different phosphorescent molecules to obtain a multicolor, long-afterglow RTP material. CS can emit RTP with a lifetime of 48 ms. In addition, CS is rich in amino groups, and grafting different phosphorescent molecules onto CS by an amidation reaction can modulate it to emit different colors of phosphorescence and obtain a series of colorful CS derivatives. The obtained polymer films also have ultra-long RTP due to the good film-forming ability. In addition, one of the CS derivatives selected with α-cyclodextrin is used to construct RTP materials with lifetimes of up to seconds. The host-guest interactions are used to suppress nonradiative relaxation and build crystalline domains, thus synergistically enhancing the RTP. Interestingly, the RTP properties of the CS derivative films are extremely sensitive to water and heat stimuli, because water broke the hydrogen bonds between adjacent CS molecules and thus altered the rigid environment in the material. Finally, they can be used as a stimuli-responsive ink and for monitoring environmental humidity.
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Epstein-Barr virus (EBV) was the first tumor virus discovered in humans and can cause various types of tumors. Molecular classification suggests that EBV-associated gastric cancer (EBVaGC) is a unique subtype of gastric cancer.EBV was also the first virus found to encode its own microRNAs. However, the functions of many miRNAs remain unknown. This study investigated the roles and targets of miR-BART2-5p (BART2-5p) and miR-BART11-5p (BART11-5p) in EBVaGC. The expression of RB and p21 in EBVaGC and EBV negative GC (EBVnGC) cells was evaluated by western blotting. Expression of BART2-5p and BART11-5p in EBVaGC cells was evaluated by droplet digital PCR. The effects of BART2-5p or BART11-5p and their potential mechanisms were further investigated using cell counting kit-8, colony formation assay, flow cytometry analysis, and transwell assay. BART2-5p and BART11-5p were abundantly expressed and RB and p21 were downregulated in EBVaGC cells. BART2-5p regulates RB and p21 expression by directly targeting them. BART11-5p regulates RB expression by directly targeting RB. Both BART2-5p and BART11-5p promoted proliferation and migration of gastric cancer cells, while inhibiting apoptosis and promoting S-phase arrest of the cell cycle. Thus, BART2-5p and BART11-5p play important roles in promoting proliferation and migration, and inhibiting apoptosis in EBVaGC by targeting RB and p21, thus providing new potential therapeutic targets for EBVaGC.
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Inibidor de Quinase Dependente de Ciclina p21 , Infecções por Vírus Epstein-Barr , MicroRNAs , RNA Viral , Proteínas de Ligação a Retinoblastoma , Neoplasias Gástricas , Humanos , Apoptose , Carcinoma/metabolismo , Carcinoma/patologia , Proliferação de Células , Infecções por Vírus Epstein-Barr/genética , Herpesvirus Humano 4/genética , MicroRNAs/metabolismo , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , RNA Viral/metabolismo , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Proteínas de Ligação a Retinoblastoma/metabolismoRESUMO
The RPMS1 gene is the only member of the BamHI-A rightward transcripts (BARTs) family for which a full-length complementary DNA has been identified, and RPMS1 transcript has been confirmed in many Epstein-Barr virus (EBV)-positive malignancies. However, the effects of sequence variations of RPMS1 in hematological malignancies and their biological significance are unclear. To explore the association between RPMS1 gene variations and hematological malignancy, the RPMS1 gene of 391 EBV-positive samples from patients with EBV-positive leukemia, myelodysplastic syndromes and lymphoma in northern China were sequenced. On the basis of phylogenetic tree and mutation characteristics of RPMS1, all the sequences were divided into five major types: RPMS1-A, RPMS1-B, RPMS1-C, RPMS1-E, and RPMS1-F. RPMS1-A type, similar to the prototype B95-8, was identified in 71.87% (281/391) of samples and was the major type in all subpopulations. The frequency of RPMS1-F type was significantly higher in all malignant hematopathy groups than in healthy donors. The Hodgkin lymphoma group contained more RPMS1-F than other malignant hematopathy groups, and acute myeloid leukemia contained more RPMS1-C type than other malignant hematopathy groups. Therefore, RPMS1-A is the main type of RPMS1 gene in northern China, and RPMS1-F may be associated with hematologic malignancies.
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Infecções por Vírus Epstein-Barr , Doença de Hodgkin , Humanos , Herpesvirus Humano 4/genética , Infecções por Vírus Epstein-Barr/complicações , Filogenia , Análise de Sequência , ChinaRESUMO
Despite the notable progress made in recent years, the understanding of the genetic control of gonadal sex differentiation and asymmetrical ovariogenesis in chicken during embryonic development remains incomplete. This study aimed to identify potential key genes and speculate about the mechanisms associated with ovary and testis development via an analysis of the results of PacBio and Illumina transcriptome sequencing of embryonic chicken gonads at the initiation of sexual differentiation (E4.5, E5.5, and E6.5). PacBio sequencing detected 328 and 233 significantly up-regulated transcript isoforms in females and males at E4.5, respectively. Illumina sequencing detected 95, 296 and 445 DEGs at E4.5, E5.5, and E6.5, respectively. Moreover, both sexes showed asymmetrical expression in gonads, and more DEGs were detected on the left side. There were 12 DEGs involved in cell proliferation shared between males and females in the left gonads. GO analysis suggested that coagulation pathways may be involved in the degradation of the right gonad in females and that blood oxygen transport pathways may be involved in preventing the degradation of the right gonad in males. These results provide a comprehensive expression profile of chicken embryo gonads at the initiation of sexual differentiation, which can serve as a theoretical basis for further understanding the mechanism of bird sex determination and its evolutionary process.
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Galinhas , Testículo , Feminino , Masculino , Animais , Embrião de Galinha , Galinhas/genética , Testículo/metabolismo , Gônadas/metabolismo , Ovário/metabolismo , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica no DesenvolvimentoRESUMO
Bacterial wilt, caused by Ralstonia solanacearum, one of the most destructive phytopathogens, leads to significant annual crop yield losses. Type III effectors (T3Es) mainly contribute to the virulence of R. solanacearum, usually by targeting immune-related proteins. Here, we clarified the effect of a novel E3 ubiquitin ligase (NEL) T3E, RipAW, from R. solanacearum on pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and further explored its action mechanism. In the susceptible host Arabidopsis thaliana, we monitored the expression of PTI marker genes, flg22-induced ROS burst, and callose deposition in RipAW- and RipAWC177A-transgenic plants. Our results demonstrated that RipAW suppressed host PTI in an NEL-dependent manner. By Split-Luciferase Complementation, Bimolecular Fluorescent Complimentary, and Co-Immunoprecipitation assays, we further showed that RipAW associated with three crucial components of the immune receptor complex, namely FLS2, XLG2, and BIK1. Furthermore, RipAW elevated the ubiquitination levels of FLS2, XLG2, and BIK1, accelerating their degradation via the 26S proteasome pathway. Additionally, co-expression of FLS2, XLG2, or BIK1 with RipAW partially but significantly restored the RipAW-suppressed ROS burst, confirming the involvement of the immune receptor complex in RipAW-regulated PTI. Overall, our results indicate that RipAW impairs host PTI by disrupting the immune receptor complex. Our findings provide new insights into the virulence mechanism of R. solanacearum.
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Proteínas de Arabidopsis , Arabidopsis , Ralstonia solanacearum , Complexo Antígeno-Anticorpo , Reconhecimento da Imunidade Inata , Espécies Reativas de Oxigênio , Imunoprecipitação , Receptores Imunológicos , Proteínas Serina-Treonina Quinases , Proteínas de Arabidopsis/genéticaRESUMO
A novel, molecularly imprinted, upconversion fluorescence probe (UCNP@MIFP) for sulfonamide sensing was fabricated by Pickering emulsion polymerization using UCNP@SiO2 particles as the stabilizer and sulfamethazine/sulfamerazine as the co-templates. The synthesis conditions of the UCNP@MIFP were optimized, and the synthesized probe was characterized by scanning electron microscopy, Fourier transform infrared spectrometer, thermogravimetric analyzer, and fluorescence spectrometer. The UCNP@MIFPs showed a good adsorption capacity and a fast kinetic feature for the template. The selectivity experiment revealed that the UCNP@MIFP has a broad-spectrum molecular recognition capability. Good linear relationships were obtained over the concentration range of 1-10 ng/mL for sulfamerazine, sulfamethazine, sulfathiazole, and sulfafurazole, with low limits of detection in the range of 1.37-2.35 ng/mL. The prepared UCNP@MIFP has the potential to detect four sulfonamide residues in food and environmental water.
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Despite metal-based photosensitizers showing great potential in photodynamic therapy for tumor treatment, the application of the photosensitizers is intrinsically limited by their poor cancer-targeting properties. Herein, we reported a metal-based photosensitizer-bacteria hybrid, Ir-HEcN, via covalent labeling of an iridium(III) photosensitizer to the surface of genetically engineered bacteria. Due to its intrinsic self-propelled motility and hypoxia tropism, Ir-HEcN selectively targets and penetrates deeply into tumor tissues. Importantly, Ir-HEcN is capable of inducing pyroptosis and immunogenic cell death of tumor cells under irradiation, thereby remarkably evoking anti-tumor innate and adaptive immune responses in vivo and leading to the regression of solid tumors via combinational photodynamic therapy and immunotherapy. To the best of our knowledge, Ir-HEcN is the first metal complex decorated bacteria for enhanced photodynamic immunotherapy.
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A quadrupolar compound Pyr-BA with two pyrrole-type nitrogen atoms doped externally was prepared in this work. In high contrast with other π ionic radicals, its cationic radical Pyr-BAâ + undergoes unusual symmetry-broken charge separation (SB-CS), generating the mixed valence complex of Pyr-BA+1-q â â â Pyr-BA+1+q , where q is the degree of charge transfer. Variable-temperature (VT) single-crystal analysis, absorption and EPR experiments all confirmed that aggregation and lower temperature would help to facilitate this SB-CS process. Gibbs energy calculations and gauge-including magnetically induced current simulation both validate that, for Pyr-BAâ + , SB-CS behavior is more favorable than the conventional dimerization mode. To the best of our knowledge, this is the first study that shows solid single-crystal evidence for spontaneous SB-CS between identical ionic radicals. Such a unique phenomenon is of great significance both in terms of fundamental aspects and uncharted material science.
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CRISPR-associated (Cas) protein systems have been increasingly incorporated in nucleic-acid diagnosis. CRISPR/Cas12a can cleave single-stranded DNA (ssDNA) after being guided to the target double-stranded DNA (dsDNA) with crRNA, making it a specific tool for dsDNA detection. Assisted by nucleic acid preamplification, CRISPR/Cas12a enables dsDNA detection at the attomolar level. However, such mandatory preamplification in CRISPR/Cas12a also accompanies the extra step of transferring preamplification products into the CRISPR/Cas12a system, which is not only cumbersome and time-consuming but also induces the risk of cross-contamination. Herein, we demonstrate a multiplex-crRNA strategy to enhance the sensitivity of the CRISPR/Cas12a system without any preamplification. This multiplex-crRNA strategy harnesses multiple sequences of crRNA which target different regions of the same dsDNA substrate in the same CRISPR/Cas12a system. Therefore, detection signals are accumulated without amplification, which augments the conventional detection limit. For application demonstration, the B646L gene from the African swine fever virus (ASFV), which is a dsDNA virus, is exemplified. The detection limit of the multiplex-crRNA system can be improved to â¼1 picomolar (pM) without amplification, which is â¼64 times stronger than the conventional single-crRNA system. The multiplex-crRNA system presented in this study, with slight modifications, can be generalized to other biosensing settings where preamplification is not readily available.
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Vírus da Febre Suína Africana , Vírus da Febre Suína Africana/genética , Animais , Sistemas CRISPR-Cas/genética , DNA/genética , DNA de Cadeia Simples , Técnicas de Amplificação de Ácido Nucleico , SuínosRESUMO
To investigate the role of miR-BART4-3p in EBV-associated gastric cancer (EBVaGC) and its regulation of cell proliferation, apoptosis, and migration by targeting AXL in GC. Quantitative real-time PCR and western blot were used to detect the expression of AXL. The methylation status of AXL gene promoter region was determined by bisulfite sequencing PCR. Luciferase reporter assay was used to detect whether miR-BART4-3p targets AXL. The key molecules of EMT and PI3K/AKT pathway were used to examine by western blot. CCK8, Transwell, and flow cytometry were used to detect the phenotypic gastric cancer cells after interference with AXL and miR-BART4-3p. EBV infection inhibited the expression of AXL in GC cells and the inhibition was not caused by the change of promoter methylation status. MiR-BART4-3p directly targeted AXL. Moreover, both inhibition of miR-BART4-3p and AXL inhibited cell proliferation and migration and promoted cell apoptosis. In addition, E-cadherin, Vimentin, ZEB1, and p-AKT were found to be the downstream molecules of the miR-BART4-3p/AXL pathway. The change of promoter methylation status was not the reason for the downregulation of AXL expression in EBV-positive cells. MiR-BART4-3p may inhibit the proliferation and migration and promote apoptosis of GC cells by directly targeting AXL.
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Carcinoma , Infecções por Vírus Epstein-Barr , MicroRNAs , Proteínas Proto-Oncogênicas , Receptores Proteína Tirosina Quinases , Neoplasias Gástricas , Apoptose/genética , Carcinoma/genética , Carcinoma/virologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Infecções por Vírus Epstein-Barr/genética , Regulação Neoplásica da Expressão Gênica , Herpesvirus Humano 4 , Humanos , MicroRNAs/genética , Fosfatidilinositol 3-Quinases/genética , Fosfatidilinositol 3-Quinases/metabolismo , Neoplasias Gástricas/genética , Neoplasias Gástricas/virologia , Receptor Tirosina Quinase AxlRESUMO
The genetic diversity and population structures of five Chongqing local chicken populations were investigated using by 24 microsatellite markers. Results revealed that the mean number of alleles (NA) ranged from 7.08 (Daninghe chicken, DN) to 8.46 (Nanchuan chicken, NC). The highest observed heterozygosity (HO) and expected heterozygosity (HE) were observed in DN (HO = 0.7252; HE = 0.7409) and the lowest HO and HE were observed in XS (Xiushan native chicken [XS], HO = 0.5910 and HE = 0.6697). The inbreeding coefficient (FIS) within population ranged from 0.022 (DN) to 0.119 (XS). Among the 24 microsatellite markers, four loci (MCW0111, MCW0016, ADL0278, and MCW0104) deviated from the Hardy-Weinberg equilibrium in all the studied populations. The results of population polygenetic analysis based on Nei's genetic distance and STRUCTURE software showed that the clustering of the five populations was incomplete consistent with geographical distribution. Moreover, a large number of gene flows were widespread among different populations, suggesting that genetic material exchanges occurred due to human activities and migration which was also verified by PCoA. In summary, this study preliminarily showed that Chongqing local chicken populations had rich genetic diversity and remarkable genetic divergence, but still high risk in conversion. These findings would be useful to the management of conservation strategies and the utilization of local chicken populations in further.
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Galinhas , Variação Genética , Humanos , Animais , Galinhas/genética , Filogenia , Variação Genética/genética , Repetições de Microssatélites/genética , AlelosRESUMO
Torreya grandis is an evergreen plant endemic of China and widely grown in Southern China. Its fruit is a precious nut in China, rich in vitamins and minerals, can be directly eaten, can also be used as medicinal plants with functions of lowering blood lipids and softening blood vessels (Wang 2022). From 2018 to 2020, typical root rot symptoms of Torreya grandis was found in plantations in Huangshan and surrounding areas of Huangshan, Anhui province, China. About 15 to 32% of root rot disease incidence was recorded at the plantation. Diseased plants were observed with symptoms such as yellow to brownish leaves without lesions and later drying, and rotten roots looked dark brown while the roots of heathy plants showed white, and eventually leading to the death of the diseased plant. The root rot symptomatic plants were collected in June of 2020. Tissues were cut to the length of 0.3 to 0.5 cm, then surface sterilized by 2% sodium hypochlorite for 2 min and 75% alcohol for 1 min, rinsed three times in sterile distilled water, and placed on potato dextrose agar (PDA) and incubated at 25â for 5 to 9 days. Eight isolates with similar morphology were isolated from single spores. On PDA, the isolates produced abundant aerial white mycelia with septation and turned violet to dark pink on the reverse side of the culture. Morphological characteristic was determined using a pure culture grown on synthetic low nutrient agar (SNA). Two types of conidia, microconidia and macroconidia, were observed on SNA. Macroconidia were long and slender, usually 3 to 5 septate, measuring 2.7 to 4.3 × 22.3 to 49.6 µm (n=30), and narrowed at the both ends. Microconidia were abundant, oval, clavate or ovate, zero to one septate and measured 1.6 to 3.9 × 4.4 to 13.0 µm (n=50). According to the culture and conidial characteristics, the isolates were tentatively identified as Fusarium species (Leslie and Summerell 2006). Four isolates were random selected for molecular identification. The general primers ITS1/ITS4 for internal transcribed spacer (ITS) (White et al. 1990), EF1/EF2 for translation elongation factor (TEF1) (O'Donnell et al. 1998), 5F2/7cR for the second largest subunit of RNA polymerase â ¡(RPB2) (O'Donnell et al., 2007), H3-1a/H3-1b for Histone H3 (Jacobs et al., 2010), F5/R8 for subunits 1 of DNA-directed RNA polymerase â ¡ (RPB1) (O'Donnell et al. 2010) and MS3F/MS3R for mitochondrial small subunit (mtSSU) (Stenglein et al. 2010) were amplified, respectively. The products were sequenced and deposited in GenBank with accession numbers of MW350689, MW029444, ON077156, ON077158, ON077157, ON054432, respectively. Blast analysis showed 99.40 to 100% sequence homology with known F. fujikuroi isolates. A phylogenetic analysis based on the concatenated sequences clustered from the combined datasets (TEF1, RPB2, Histone H3, RPB1 and mtSSU) revealed the isolate most closely related to the F. fujikuroi (100% bootstrap). Fifteen 2-year-old healthy plants of Torreya grandis were selected for the pathogenicity test. A conidial suspension (1×106 conidia/ml) was prepared by collecting spores from 10-day-old cultures on PDA. The root of each plants inoculated with 200 ml of a 106 conidia/ml suspension, and the five control plants inoculated with sterilized water. The plants were incubated in green house with 25â (14 h light)/22â (10 h dark) at 85% humidity. Two weeks later, 100% of artificially inoculated plants showed the same symptoms similar to those observed in the plantation, like yellow leaves, dark brown and rotten roots, meanwhile, the roots of control plants displayed healthy. From symptomatic roots, the pathogen was reisolated which satisfying Koch's postulates. F. fujikuroi causes root rot of soybean and Reineckia carnea (Detranaltes et al. 2021, Sun et al. 2018).To the best of our knowledge, this is the first report of F. fujikuroi causing root rot of Torreya grandis in China.
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OBJECTIVES: This meta-analysis aimed to evaluate quantitative sensory testing (QST) evidence for pain processing in patients with the muscle pain subtype of temporomandibular disorders (mTMD). MATERIALS AND METHODS: A comprehensive systematic electronic search strategy was performed in online literature databases. All full-text observational studies published up to July 2021 with the aim of investigating pain sensitization in humans with mTMD using QST measures were eligible for inclusion. Meta-analysis of QST data was performed using a random effects model, which included results comparing patients with mTMD to healthy controls, and standard mean difference (SMD) results were analyzed. RESULTS: Twelve studies with 732 participants (371 patients with mTMD and 361 healthy controls) were analyzed following screening and quality appraisal. Compared with healthy controls, patients with mTMD had significantly lower pressure pain threshold (SMD - 1.10, 95% confidence interval [CI] - 1.52 to - 0.68) with high heterogeneity (Tau2 = 0.61, I2 = 86%), and significantly lower mechanical pain threshold (SMD - 0.64, 95% CI - 0.95 to - 0.32) with no heterogeneity (Tau2 = 0.00, I2 = 0%). No difference was observed in the cold pain threshold (SMD 0.16, 95% CI - 0.13 to 0.45), heat pain threshold (SMD - 0.13, 95% CI - 0.40 to 0.15), and wind-up ratio (SMD 0.63, 95% CI - 0.11 to 1.38) between patients with mTMD and healthy controls. Other QST parameters were also discussed. CONCLUSIONS: The study results suggest that the pain processing of deep tissues is likely sensitized in mTMD and calls for more QST studies with standard procedures to reduce inter-study heterogeneity. CLINICAL RELEVANCE: The major findings of this meta-analysis support using PPT to examine the pain processing in patients with mTMD in clinical scenario.
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Mialgia , Transtornos da Articulação Temporomandibular , Humanos , Medição da Dor , Limiar da DorRESUMO
BACKGROUND: Knowledge of gingival thickness (GT) and alveolar crest thickness (ACT) is essential when performing surgical and non-surgical procedures in the maxillary anterior teeth region. This study aimed at evaluating the GT and ACT in the maxillary anterior teeth region using 15-MHz B-mode Ultrasonic (US). METHODS: A total of 300 teeth from 50 healthy participants, comprising 25 women and 25 men, aged between 18 and 35 years were analyzed. We measured labial periodontal tissue structures of maxillary anterior teeth, including GT and ACT, at 3 mm apical to the gingival margin (GT3) and the crestal level, respectively. The GT and ACT measurements were correlated. RESULTS: The mean labial GT3 of the maxillary central incisors, lateral incisors, and canines were 1.24 ± 0.03 mm, 1.21 ± 0.03 mm and 1.11 ± 0.03 mm, respectively. Canine GT3 was significantly thin than those in the central and lateral incisors (P < 0.05). With regards to labial ACT, we recorded 0.79 ± 0.03 mm, 0.76 ± 0.02 mm and 0.73 ± 0.02 mm for maxillary central incisors, lateral incisors and canines, respectively. There were no significant differences in ACT of maxillary anterior teeth (P > 0.05). GT3 of men was greater than that of women (P < 0.05). In addition, GT and ACT were positively correlated (r = 0.32, P < 0.01). CONCLUSION: 15-MHz B-mode US is an effective tool for measuring labial GT and ACT of anterior teeth. There are sex-associated differences in GT3 and the correlation between the GT3 and ACT of anterior teeth is moderately positive.
Assuntos
Dente Canino , Maxila , Adolescente , Adulto , Tomografia Computadorizada de Feixe Cônico , Dente Canino/diagnóstico por imagem , Feminino , Gengiva/diagnóstico por imagem , Humanos , Incisivo/diagnóstico por imagem , Masculino , Maxila/diagnóstico por imagem , Ultrassonografia , Adulto JovemRESUMO
BACKGROUND: The global burden of hepatitis B virus (HBV) infection in terms of morbidity and mortality is immense. Novel treatments that can induce a protective immune response are urgently needed to effectively control the HBV epidemic and eventually eradicate chronic HBV infection. METHODS: We designed and evaluated an HBV therapeutic vaccine consisting of a novel Toll-like receptor 7 (TLR7) agonist T7-EA, an Alum adjuvant and a recombinant HBsAg protein. We used RNA-seq, ELISA and hTLR7/8 reporting assays to characterize T7-EA in vitro and real-time PCR to evaluate the tissue-retention characteristics in vivo. To evaluate the adjuvant potential, we administrated T7-EA intraperitoneally in a formulation with an Alum adjuvant and HBsAg in normal and HBV mice, then, we evaluated the HBsAg-specific immune responses by ELISA and Elispot assays. RESULTS: T7-EA acted as an hTLR7-specific agonist and induced a similar gene expression pattern as an unmodified TLR7 ligand when Raw 264.7 cells were exposed to T7-EA; however, T7-EA was more potent than the unmodified TLR7 ligand. In vivo studies showed that T7-EA had tissue-retaining activity with stimulating local cytokine and chemokine expression for up to 7 days. T7-EA could induce Th1-type immune responses, as evidenced by an increased HBsAg-specific IgG2a titer and a T-cell response in normal mice compared to mice received traditional Alum-adjuvant HBV vaccine. Importantly, T7-EA could break immune tolerance and induce persistent HBsAg-specific antibody and T-cell responses in an HBV mouse model. CONCLUSIONS: T7-EA might be a candidate adjuvant in a prophylactic and therapeutic HBV vaccine.