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1.
bioRxiv ; 2024 Apr 13.
Artigo em Inglês | MEDLINE | ID: mdl-38645232

RESUMO

Adenocarcinomas from multiple tissues can converge to treatment-resistant small cell neuroendocrine (SCN) cancers comprised of ASCL1, POU2F3, NEUROD1, and YAP1 subtypes. We investigated how mitochondrial metabolism influences SCN cancer (SCNC) progression. Extensive bioinformatics analyses encompassing thousands of patient tumors and human cancer cell lines uncovered enhanced expression of PGC-1α, a potent regulator of mitochondrial oxidative phosphorylation (OXPHOS), across several SCNC types. PGC-1α correlated tightly with increased expression of the lineage marker ASCL1 through a positive feedback mechanism. Analyses using a human prostate tissue-based SCN transformation system showed that the ASCL1 subtype has heightened PGC-1α expression and OXPHOS activity. PGC-1α inhibition diminished OXPHOS, reduced SCNC cell proliferation, and blocked SCN prostate tumor formation. PGC-1α overexpression enhanced OXPHOS, tripled the SCN prostate tumor formation rate, and promoted commitment to the ASCL1 lineage. These findings reveal the metabolic heterogeneity among SCNC subtypes and identify PGC-1α-induced OXPHOS as a regulator of SCNC lineage plasticity.

3.
Proc Natl Acad Sci U S A ; 121(14): e2320442121, 2024 Apr 02.
Artigo em Inglês | MEDLINE | ID: mdl-38536748

RESUMO

The ability to selectively bind to antigenic peptides and secrete effector molecules can define rare and low-affinity populations of cells with therapeutic potential in emerging T cell receptor (TCR) immunotherapies. We leverage cavity-containing hydrogel microparticles, called nanovials, each coated with peptide-major histocompatibility complex (pMHC) monomers to isolate antigen-reactive T cells. T cells are captured and activated by pMHCs inducing the secretion of effector molecules including IFN-γ and granzyme B that are accumulated on nanovials, allowing sorting based on both binding and function. The TCRs of sorted cells on nanovials are sequenced, recovering paired αß-chains using microfluidic emulsion-based single-cell sequencing. By labeling nanovials having different pMHCs with unique oligonucleotide-barcodes and secretions with oligo-barcoded detection antibodies, we could accurately link TCR sequences to specific targets and rank each TCR based on the corresponding cell's secretion level. Using the technique, we identified an expanded repertoire of functional TCRs targeting viral antigens with high specificity and found rare TCRs with activity against cancer-specific splicing-enhanced epitopes.


Assuntos
Receptores de Antígenos de Linfócitos T , Linfócitos T , Peptídeos/química , Antígenos de Histocompatibilidade/química , Antígenos
4.
J Mol Neurosci ; 74(1): 28, 2024 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-38441703

RESUMO

Mounting evidence suggests a significant correlation between depressive disorders and neurodegenerative conditions, encompassing Alzheimer's disease and Parkinson's disease (PD). Depression represents a substantial non-motor manifestation frequently identified in individuals with PD, posing a significant threat to patients' overall well-being and necessitating the implementation of effective management strategies. Despite its high prevalence, impacting over 40% of PD patients, the precise cellular and molecular mechanisms underlying depression and its relationship to dopaminergic system degeneration remain largely ambiguous. In this study, we presented our findings demonstrating distinct characteristics of cortical astrocytes in PD patients compared to reactivated glial cells in the substantia nigra. We identified a subset of differentially expressed genes associated with depressive disorders from PD-associated cortical astrocytes. Furthermore, we uncovered the potential involvement of the hypoxia signaling in driving cortical astrocytic dysfunctions. Through a comprehensive investigation utilizing transcriptome and chromatin accessibility analyses on cultured human astrocytes, we revealed that hypoxic treatment could induce similar expression changes observed in cortex from PD patients. Additionally, we provided evidence that activation of the HIF-1 signaling pathway suppressed the expression of key components of mitochondrial ribosomes and electron transport chain proteins COX2 and CYTB, resulting in abnormal mitochondrial membrane potential. Our results underscore the potential impact of glial metabolic abnormalities on the development of depressive disorders associated with Parkinson's disease.


Assuntos
Astrócitos , Doença de Parkinson , Humanos , Doença de Parkinson/genética , Depressão/etiologia , Neuroglia , Hipóxia
5.
Proc Natl Acad Sci U S A ; 120(47): e2312374120, 2023 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-37963244

RESUMO

CAR (chimeric antigen receptor) T cell therapy has shown clinical success in treating hematological malignancies, but its treatment of solid tumors has been limited. One major challenge is on-target, off-tumor toxicity, where CAR T cells also damage normal tissues that express the targeted antigen. To reduce this detrimental side-effect, Boolean-logic gates like AND-NOT gates have utilized an inhibitory CAR (iCAR) to specifically curb CAR T cell activity at selected nonmalignant tissue sites. However, the strategy seems inefficient, requiring high levels of iCAR and its target antigen for inhibition. Using a TROP2-targeting iCAR with a single PD1 inhibitory domain to inhibit a CEACAM5-targeting CAR (CEACAR), we observed that the inefficiency was due to a kinetic delay in iCAR inhibition of cytotoxicity. To improve iCAR efficiency, we modified three features of the iCAR-the avidity, the affinity, and the intracellular signaling domains. Increasing the avidity but not the affinity of the iCAR led to significant reductions in the delay. iCARs containing twelve different inhibitory signaling domains were screened for improved inhibition, and three domains (BTLA, LAIR-1, and SIGLEC-9) each suppressed CAR T function but did not enhance inhibitory kinetics. When inhibitory domains of LAIR-1 or SIGLEC-9 were combined with PD-1 into a single dual-inhibitory domain iCAR (DiCARs) and tested with the CEACAR, inhibition efficiency improved as evidenced by a significant reduction in the inhibitory delay. These data indicate that a delicate balance between CAR and iCAR signaling strength and kinetics must be achieved to regulate AND-NOT gate CAR T cell selectivity.


Assuntos
Neoplasias , Receptores de Antígenos Quiméricos , Humanos , Receptores de Antígenos Quiméricos/genética , Linfócitos T , Complexo Ferro-Dextran , Imunoterapia Adotiva , Lectinas Semelhantes a Imunoglobulina de Ligação ao Ácido Siálico
6.
Cancer Cell ; 41(12): 2066-2082.e9, 2023 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-37995683

RESUMO

Trans-differentiation from an adenocarcinoma to a small cell neuroendocrine state is associated with therapy resistance in multiple cancer types. To gain insight into the underlying molecular events of the trans-differentiation, we perform a multi-omics time course analysis of a pan-small cell neuroendocrine cancer model (termed PARCB), a forward genetic transformation using human prostate basal cells and identify a shared developmental, arc-like, and entropy-high trajectory among all transformation model replicates. Further mapping with single cell resolution reveals two distinct lineages defined by mutually exclusive expression of ASCL1 or ASCL2. Temporal regulation by groups of transcription factors across developmental stages reveals that cellular reprogramming precedes the induction of neuronal programs. TFAP4 and ASCL1/2 feedback are identified as potential regulators of ASCL1 and ASCL2 expression. Our study provides temporal transcriptional patterns and uncovers pan-tissue parallels between prostate and lung cancers, as well as connections to normal neuroendocrine cell states.


Assuntos
Carcinoma de Células Pequenas , Neoplasias Pulmonares , Neoplasias da Próstata , Carcinoma de Pequenas Células do Pulmão , Masculino , Humanos , Neoplasias Pulmonares/genética , Carcinoma de Células Pequenas/genética , Fatores de Transcrição/genética , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , Transdiferenciação Celular/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Regulação Neoplásica da Expressão Gênica , Linhagem Celular Tumoral , Carcinoma de Pequenas Células do Pulmão/genética
7.
Proc Natl Acad Sci U S A ; 120(28): e2220190120, 2023 07 11.
Artigo em Inglês | MEDLINE | ID: mdl-37399401

RESUMO

The MYC proto-oncogene contributes to the pathogenesis of more than half of human cancers. Malignant transformation by MYC transcriptionally up-regulates the core pre-mRNA splicing machinery and causes misregulation of alternative splicing. However, our understanding of how splicing changes are directed by MYC is limited. We performed a signaling pathway-guided splicing analysis to identify MYC-dependent splicing events. These included an HRAS cassette exon repressed by MYC across multiple tumor types. To molecularly dissect the regulation of this HRAS exon, we used antisense oligonucleotide tiling to identify splicing enhancers and silencers in its flanking introns. RNA-binding motif prediction indicated multiple binding sites for hnRNP H and hnRNP F within these cis-regulatory elements. Using siRNA knockdown and cDNA expression, we found that both hnRNP H and F activate the HRAS cassette exon. Mutagenesis and targeted RNA immunoprecipitation implicate two downstream G-rich elements in this splicing activation. Analyses of ENCODE RNA-seq datasets confirmed hnRNP H regulation of HRAS splicing. Analyses of RNA-seq datasets across multiple cancers showed a negative correlation of HNRNPH gene expression with MYC hallmark enrichment, consistent with the effect of hnRNP H on HRAS splicing. Interestingly, HNRNPF expression showed a positive correlation with MYC hallmarks and thus was not consistent with the observed effects of hnRNP F. Loss of hnRNP H/F altered cell cycle progression and induced apoptosis in the PC3 prostate cancer cell line. Collectively, our results reveal mechanisms for MYC-dependent regulation of splicing and point to possible therapeutic targets in prostate cancers.


Assuntos
Ribonucleoproteínas Nucleares Heterogêneas Grupo F-H , Neoplasias da Próstata , Masculino , Humanos , Ribonucleoproteínas Nucleares Heterogêneas Grupo F-H/genética , Ribonucleoproteínas Nucleares Heterogêneas Grupo F-H/metabolismo , Precursores de RNA/genética , Precursores de RNA/metabolismo , Splicing de RNA/genética , Proteínas de Ligação a RNA/metabolismo , Éxons/genética , Processamento Alternativo/genética , Neoplasias da Próstata/genética , Proteínas Proto-Oncogênicas p21(ras)/genética , Proteínas Proto-Oncogênicas p21(ras)/metabolismo
8.
Proc Natl Acad Sci U S A ; 120(21): e2221116120, 2023 05 23.
Artigo em Inglês | MEDLINE | ID: mdl-37192158

RESUMO

Alternative splicing (AS) is prevalent in cancer, generating an extensive but largely unexplored repertoire of novel immunotherapy targets. We describe Isoform peptides from RNA splicing for Immunotherapy target Screening (IRIS), a computational platform capable of discovering AS-derived tumor antigens (TAs) for T cell receptor (TCR) and chimeric antigen receptor T cell (CAR-T) therapies. IRIS leverages large-scale tumor and normal transcriptome data and incorporates multiple screening approaches to discover AS-derived TAs with tumor-associated or tumor-specific expression. In a proof-of-concept analysis integrating transcriptomics and immunopeptidomics data, we showed that hundreds of IRIS-predicted TCR targets are presented by human leukocyte antigen (HLA) molecules. We applied IRIS to RNA-seq data of neuroendocrine prostate cancer (NEPC). From 2,939 NEPC-associated AS events, IRIS predicted 1,651 epitopes from 808 events as potential TCR targets for two common HLA types (A*02:01 and A*03:01). A more stringent screening test prioritized 48 epitopes from 20 events with "neoantigen-like" NEPC-specific expression. Predicted epitopes are often encoded by microexons of ≤30 nucleotides. To validate the immunogenicity and T cell recognition of IRIS-predicted TCR epitopes, we performed in vitro T cell priming in combination with single-cell TCR sequencing. Seven TCRs transduced into human peripheral blood mononuclear cells (PBMCs) showed high activity against individual IRIS-predicted epitopes, providing strong evidence of isolated TCRs reactive to AS-derived peptides. One selected TCR showed efficient cytotoxicity against target cells expressing the target peptide. Our study illustrates the contribution of AS to the TA repertoire of cancer cells and demonstrates the utility of IRIS for discovering AS-derived TAs and expanding cancer immunotherapies.


Assuntos
Neoplasias , Precursores de RNA , Masculino , Humanos , Precursores de RNA/metabolismo , Processamento Alternativo , Leucócitos Mononucleares/metabolismo , Receptores de Antígenos de Linfócitos T , Epitopos de Linfócito T , Imunoterapia , Antígenos de Neoplasias , Peptídeos/metabolismo , Neoplasias/genética , Neoplasias/terapia
9.
bioRxiv ; 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36711524

RESUMO

The ability to selectively bind to antigenic peptides and secrete cytokines can define populations of cells with therapeutic potential in emerging T cell receptor (TCR) immunotherapies. We leverage cavity-containing hydrogel microparticles, called nanovials, each coated with millions of peptide-major histocompatibility complex (pMHC) monomers to isolate antigen-reactive T cells. T cells are captured and activated by pMHCs and secrete cytokines on nanovials, allowing sorting based on both affinity and function. The TCRs of sorted cells on nanovials are sequenced, recovering paired αß-chains using microfluidic emulsion-based single-cell sequencing. By labeling nanovials having different pMHCs with unique oligonucleotide-barcodes we could link TCR sequence to targets with 100% accuracy. We identified with high specificity an expanded repertoire of functional TCRs targeting viral antigens compared to standard techniques.

10.
Trop Med Infect Dis ; 7(12)2022 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-36548668

RESUMO

Ticks can carry and transmit a large number of pathogens, including bacteria, viruses and protozoa, posing a huge threat to human health and animal husbandry. Previous investigations have shown that the dominant species of ticks in Shanghai are Haemaphysalis flava and Haemaphysalis longicornis. However, no relevant investigations and research have been carried out in recent decades. Therefore, we investigated the bacterial communities and tick-borne pathogens (TBPs) in Haemaphysalis spp. from Shanghai, China. Ixodid ticks were collected from 18 sites in Shanghai, China, and identified using morphological and molecular methods. The V3-V4 hypervariable regions of the bacterial 16S rRNA gene were amplified from the pooled tick DNA samples and subject to metagenomic analysis. The microbial diversity in the tick samples was estimated using the alpha diversity that includes the observed species index and Shannon index. The Unifrac distance matrix as determined using the QIIME software was used for unweighted Unifrac Principal coordinates analysis (PCoA). Individual tick DNA samples were screened with genus-specific or group-specific nested polymerase chain reaction (PCR) for these TBPs and combined with a sequencing assay to confirm the results of the V3-V4 hypervariable regions of the bacterial 16S rRNA gene. We found H. flava and H. longicornis to be the dominant species of ticks in Shanghai in this study. Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria are the main bacterial communities of Haemaphysalis spp. The total species abundances of Proteobacteria, Firmicutes and Bacteroidetes, are 48.8%, 20.8% and 18.1%, respectively. At the level of genus analysis, H. longicornis and H. flava carried at least 946 genera of bacteria. The bacteria with high abundance include Lactobacillus, Coxiella, Rickettsia and Muribaculaceae. Additionally, Rickettsia rickettsii, Rickettsia japonica, Candidatus Rickettsia jingxinensis, Anaplasma bovis, Ehrlichia ewingii, Ehrlichia chaffeensis, Coxiella spp. and Coxiella-like endosymbiont were detected in Haemaphysalis spp. from Shanghai, China. This study is the first report of bacterial communities and the prevalence of some main pathogens in Haemaphysalis spp. from Shanghai, China, and may provide insights and evidence for bacterial communities and the prevalence of the main pathogen in ticks. This study also indicates that people and other animals in Shanghai, China, are exposed to several TBPs.

11.
World J Clin Cases ; 10(31): 11579-11584, 2022 Nov 06.
Artigo em Inglês | MEDLINE | ID: mdl-36387803

RESUMO

BACKGROUND: Inadequate volume of future liver remnant (FLR) is a major challenge for hepatobiliary surgeons treating large or multiple liver tumors. As an alternative to associating liver partition and portal vein ligation (ALPPS) for staged hepatectomy and liver venous deprivation (LVD) using stage 1 interventional radiology for vascular embolization combined with stage 2 open liver resection have been used. CASE SUMMARY: A novel modified LVD technique was performed in a patient with pancreatic neuroendocrine tumor with liver metastases by using stage 1 laparoscopic ligation of the right hepatic vein, right posterior portal vein, and short hepatic veins combined with local excision of three liver metastases in the left hemiliver. The operation was followed three days later by interventional radiology to embolize an anomalous right anterior portal vein to complete LVD. A stage 2 laparoscopic right hemihepatectomy and pancreaticosplenectomy were then carried out. CONCLUSION: The minimally invasive technique promoted a rapid increase, comparable to ALPPS, in volume of the FLR after the stage 1 operation to allow the laparoscopic stage 2 resection to be performed.

12.
Proc Natl Acad Sci U S A ; 119(31): e2203410119, 2022 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-35878026

RESUMO

Tissue-specific antigens can serve as targets for adoptive T cell transfer-based cancer immunotherapy. Recognition of tumor by T cells is mediated by interaction between peptide-major histocompatibility complexes (pMHCs) and T cell receptors (TCRs). Revealing the identity of peptides bound to MHC is critical in discovering cognate TCRs and predicting potential toxicity. We performed multimodal immunopeptidomic analyses for human prostatic acid phosphatase (PAP), a well-recognized tissue antigen. Three physical methods, including mild acid elution, coimmunoprecipitation, and secreted MHC precipitation, were used to capture a thorough signature of PAP on HLA-A*02:01. Eleven PAP peptides that are potentially A*02:01-restricted were identified, including five predicted strong binders by NetMHCpan 4.0. Peripheral blood mononuclear cells (PBMCs) from more than 20 healthy donors were screened with the PAP peptides. Seven cognate TCRs were isolated which can recognize three distinct epitopes when expressed in PBMCs. One TCR shows reactivity toward cell lines expressing both full-length PAP and HLA-A*02:01. Our results show that a combined multimodal immunopeptidomic approach is productive in revealing target peptides and defining the cloned TCR sequences reactive with prostatic acid phosphatase epitopes.


Assuntos
Fosfatase Ácida , Antígenos de Neoplasias , Receptores de Antígenos de Linfócitos T , Fosfatase Ácida/metabolismo , Antígenos de Neoplasias/metabolismo , Epitopos , Antígenos HLA-A/metabolismo , Antígeno HLA-A2 , Humanos , Leucócitos Mononucleares , Neoplasias/imunologia , Peptídeos , Receptores de Antígenos de Linfócitos T/metabolismo
13.
Materials (Basel) ; 15(12)2022 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-35744298

RESUMO

This study aims to evaluate the mechanical properties of carbon fiber-reinforced reactive powder concrete (CFRPC) after exposure to cryogenic temperature. The mechanical properties of plain RPC and CFRPC with carbon fiber volume contents of 0, 0.5%, 1.0%, and 1.5% were examined after exposure to 20 °C, -5 °C, -15 °C, and -25 °C for 72 h. The effect of fiber contents and exposure temperatures on the cubic and axial compressive strength, splitting tensile strength, elastic modulus, and peak strain were systematically reported and analyzed. The results showed adding carbon fiber to RPC could significantly enhance the strength and slightly improve ductility performance. Additionally, CFRPC with 1.0% fiber content showed the best mechanical properties. The maximum increases in cubic and axial compressive strength and tensile strength were 26.0%, 25.7%, and 21.8%, the elastic modulus was 13.2%, and the peak strain was 13.0% over the plain RPC. Additionally, all mechanical properties continued to degrade with decreasing temperature. After exposure to -25 °C, the cubic, axial compressive strength, and tensile strength of CFRPC degraded to 82.2-84.9%, 80.7-87.5%, and 72.7-73.7% of the normal temperature strength, respectively. In addition, the linear relationship equation between the discount factor of each mechanical property and the temperature was established. Finally, the equation for the stress-strain ascending curve of CFRPC described by a quadratic polynomial was proposed, which fitted well with the experimental results.

14.
Cell Rep ; 37(13): 110167, 2021 12 28.
Artigo em Inglês | MEDLINE | ID: mdl-34919800

RESUMO

Cross-reactivity and direct killing of target cells remain underexplored for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2)-specific CD8+ T cells. Isolation of T cell receptors (TCRs) and overexpression in allogeneic cells allows for extensive T cell reactivity profiling. We identify SARS-CoV-2 RNA-dependent RNA polymerase (RdRp/NSP12) as highly conserved, likely due to its critical role in the virus life cycle. We perform single-cell TCRαß sequencing in human leukocyte antigen (HLA)-A∗02:01-restricted, RdRp-specific T cells from SARS-CoV-2-unexposed individuals. Human T cells expressing these TCRαß constructs kill target cell lines engineered to express full-length RdRp. Three TCR constructs recognize homologous epitopes from common cold coronaviruses, indicating CD8+ T cells can recognize evolutionarily diverse coronaviruses. Analysis of individual TCR clones may help define vaccine epitopes that can induce long-term immunity against SARS-CoV-2 and other coronaviruses.


Assuntos
RNA-Polimerase RNA-Dependente de Coronavírus/imunologia , Antígeno HLA-A2/imunologia , SARS-CoV-2/imunologia , Linfócitos T CD8-Positivos/imunologia , COVID-19/imunologia , COVID-19/terapia , Técnicas de Cultura de Células , Reações Cruzadas/imunologia , Epitopos de Linfócito T/imunologia , Antígenos HLA-A/imunologia , Antígeno HLA-A2/genética , Humanos , Epitopos Imunodominantes/imunologia , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/metabolismo , Leucócitos Mononucleares/virologia , RNA Viral/genética , Receptores de Antígenos de Linfócitos T/imunologia , Receptores de Antígenos de Linfócitos T alfa-beta/genética , Receptores de Antígenos de Linfócitos T alfa-beta/imunologia , SARS-CoV-2/patogenicidade , Glicoproteína da Espícula de Coronavírus/imunologia
15.
Cell Rep Med ; 2(11): 100449, 2021 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-34841295

RESUMO

Cell-based immunotherapy has become the new-generation cancer medicine, and "off-the-shelf" cell products that can be manufactured at large scale and distributed readily to treat patients are necessary. Invariant natural killer T (iNKT) cells are ideal cell carriers for developing allogeneic cell therapy because they are powerful immune cells targeting cancers without graft-versus-host disease (GvHD) risk. However, healthy donor blood contains extremely low numbers of endogenous iNKT cells. Here, by combining hematopoietic stem cell (HSC) gene engineering and in vitro differentiation, we generate human allogeneic HSC-engineered iNKT (AlloHSC-iNKT) cells at high yield and purity; these cells closely resemble endogenous iNKT cells, effectively target tumor cells using multiple mechanisms, and exhibit high safety and low immunogenicity. These cells can be further engineered with chimeric antigen receptor (CAR) to enhance tumor targeting or/and gene edited to ablate surface human leukocyte antigen (HLA) molecules and further reduce immunogenicity. Collectively, these preclinical studies demonstrate the feasibility and cancer therapy potential of AlloHSC-iNKT cell products and lay a foundation for their translational and clinical development.


Assuntos
Células Alógenas/imunologia , Engenharia Celular , Células-Tronco Hematopoéticas/imunologia , Imunoterapia , Células T Matadoras Naturais/imunologia , Neoplasias/imunologia , Neoplasias/terapia , Células Alógenas/metabolismo , Animais , Linhagem Celular Tumoral , Perfilação da Expressão Gênica , Antígenos HLA/metabolismo , Células-Tronco Hematopoéticas/metabolismo , Humanos , Camundongos Endogâmicos NOD , Camundongos SCID , Células T Matadoras Naturais/metabolismo , Fenótipo , Receptores de Antígenos Quiméricos/metabolismo , Transcriptoma/genética
16.
Huan Jing Ke Xue ; 42(7): 3565-3576, 2021 Jul 08.
Artigo em Chinês | MEDLINE | ID: mdl-34212683

RESUMO

This study explored the responses of soil dissolved organic matter (DOM) to the application of different types of compost using a soil sample without compost as a control. Ultraviolet and fluorescence spectrum technology and EEM-PARAFAC was used to analyze DOM structure and driving factors in soil added with different proportion of cow dung compost (SCC), food and kitchen waste compost (SFC), and sludge compost (SCC). Compared with the control group, contents of AN, NH4+-N, DOC, and SOM in soil added with compost were significantly increased, and contents of SOM and DOC increased with the increasing of compost amount. When added compost in the same proportion, contents of AN, NO3--N, and DOC in SCC and SFC were significantly higher than those in SSC, while contents of NH4+-N and SOM were higher in SSC. The results of spectral analysis showed that the structure of conjugated benzene ring, hydrophobic component, quinone group, and chromogenic component in DOM of soil added with compost were significantly increased, the transition of unsaturated organic molecule (π→π*) was more active, the molecular weight of DOM increased, and the degree of humification was enhanced. When the amount of compost added is 5%, the influence of food and kitchen waste compost on DOM structure was greatest among three types of compost. At 10% and 20%, sludge compost had the greatest impact on DOM structure. The results of EEM-PARAFAC analysis showed that the relative content of fulvic acid-like substances with low molecular in DOM of soil added with compost was increased, while the relative content of proteoid-like substances decreased. 2D-COS analysis showed that compost affected the change order of fluorescence components in DOM. SCC and SFC were as follows:proteoid-like > fulvic acid-like > humus-like; in SSC, it was fulvic acid-like > proteoid-like > humus-like. The enhance of humification and the decrease of relative content of protein-like substances in DOM were related to increased DOC and AN, the relative content of humus-like in low molecular weight was positively correlated with the content of NO3--N, and the relative content of macromolecule fulvic acid-like was increased due to the input of SOM from compost.


Assuntos
Compostagem , Solo , Substâncias Húmicas/análise , Espectrometria de Fluorescência
17.
Proc Natl Acad Sci U S A ; 118(3)2021 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-33431692

RESUMO

T cell receptors (TCRs) are generated by somatic recombination of V/D/J segments to produce up to 1015 unique sequences. Highly sensitive and specific techniques are required to isolate and identify the rare TCR sequences that respond to antigens of interest. Here, we describe the use of mRNA sequencing via cross-linker regulated intracellular phenotype (CLInt-Seq) for efficient recovery of antigen-specific TCRs in cells stained for combinations of intracellular proteins such as cytokines or transcription factors. This method enables high-throughput identification and isolation of low-frequency TCRs specific for any antigen. As a proof of principle, intracellular staining for TNFα and IFNγ identified cytomegalovirus (CMV)- and Epstein-Barr virus (EBV)-reactive TCRs with efficiencies similar to state-of-the-art peptide-MHC multimer methodology. In a separate experiment, regulatory T cells were profiled based on intracellular FOXP3 staining, demonstrating the ability to examine phenotypes based on transcription factors. We further optimized the intracellular staining conditions to use a chemically cleavable primary amine cross-linker compatible with current single-cell sequencing technology. CLInt-Seq for TNFα and IFNγ performed similarly to isolation with multimer staining for EBV-reactive TCRs. We anticipate CLInt-Seq will enable droplet-based single-cell mRNA analysis from any tissue where minor populations need to be isolated by intracellular markers.


Assuntos
Fatores de Transcrição Forkhead/genética , Interferon gama/genética , Fator de Necrose Tumoral alfa/genética , Recombinação V(D)J/genética , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/virologia , Clonagem Molecular , Citomegalovirus/imunologia , Citomegalovirus/patogenicidade , Epitopos/imunologia , Infecções por Vírus Epstein-Barr/genética , Infecções por Vírus Epstein-Barr/imunologia , Infecções por Vírus Epstein-Barr/virologia , Herpesvirus Humano 4/imunologia , Herpesvirus Humano 4/patogenicidade , Humanos , RNA Mensageiro/genética , RNA-Seq , Receptores de Antígenos de Linfócitos T/genética , Receptores de Antígenos de Linfócitos T/imunologia , Análise de Célula Única , Linfócitos T Reguladores/imunologia , Recombinação V(D)J/imunologia
18.
J Prosthet Dent ; 126(5): 703-708, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-33041074

RESUMO

STATEMENT OF PROBLEM: Because few 3D-printing resins have been specifically developed for liquid crystal display (LCD) 3D printers, mono-LCD users may use digital light processing (DLP) 3D-printing resins. However, the suitability of these resins requires evaluation. PURPOSE: The purpose of this in vitro study was to evaluate whether 3D-printing resins designed for DLP 3D printers can be used successfully in a mono-LCD 3D printer. MATERIAL AND METHODS: 3D printers based on photopolymerization and 3D-printing resin for interim restorations were used in this study. Enlighten AA temp and NextDent C&B MFH were printed from both the MiiCraft Ultra 125 and Phrozen Sonic printers, followed by postpolymerization by using the FormCure and PhrozenCure units for different times. The flexural strength and cytotoxicity of the specimens were evaluated. RESULTS: After postpolymerization treatment, the flexural strength of Enlighten AA temp and NextDent C&B MFH from both 3D printers was over the 50-MPa minimal requirement for the flexural strength of interim resins specified in the International Standards Organization (ISO) 10477 standard. With 15 minutes of FormCure and 1 minute of PhrozenCure postpolymerization, 4 material-printer combinations reached nearly 100% in cell viability. CONCLUSIONS: Interim resins designed for DLP 3D printers can be successfully used in mono-LCD 3D printers if the printed specimens are postpolymerized in a more powerful postpolymerization unit or in a less powerful postpolymerization unit for a longer time.


Assuntos
Resistência à Flexão , Cristais Líquidos , Teste de Materiais , Impressão Tridimensional , Temperatura
19.
J Prosthet Dent ; 125(3): 544.e1-544.e8, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33243474

RESUMO

STATEMENT OF PROBLEM: Information on the bond strength of milled polymethyl methacrylate interim restorations when relined with chairside reline materials is lacking. PURPOSE: The purpose of this in vitro study was to measure the shear bond strength of various combinations of 3 different chairside reline materials bonded to milled polymethyl methacrylate blocks with 3 different types of surface treatments. MATERIALS AND METHODS: Uniform blocks (10×10×22 mm) were milled from tooth-colored polymethyl methacrylate disks (Vivid PMMA; Pearson Dental Supply Co). The surface treatments tested were airborne-particle abrasion with 50-µm particle size aluminosilicate, application of acrylic resin monomer (Jet Liquid; Lang Dental Manufacturing Co) for 180 seconds, and airborne-particle abrasion with monomer application. The control groups were blocks with no surface treatment. The chairside reline materials tested were Jet acrylic resin (Jet Powder; Lang Dental Manufacturing Co), bis-acryl resin (Integrity; Dentsply Sirona), and flowable composite resin (Reveal; Bisco). All materials were applied through a Ø1.5×3-mm bonding ring. Ten specimens for each of the 12 groups were tested in a universal testing machine. Load was applied at a crosshead speed of 1 mm/min. Fracture surfaces were then analyzed for cohesive versus adhesive or mixed failure. Data were analyzed using 2-way ANOVA and Tukey-Kramer post hoc analysis (α=.05). RESULTS: The mean shear bond strength values ranged from 1.77 ±0.79 MPa to 28.49 ±5.75 MPa. ANOVA revealed that reline material (P<.05), surface treatment (P<.05), and their interactions (P<.05) significantly affected the shear bond strength among the experimental groups. The strongest combination was Jet acrylic resin applied on specimens treated with airborne-particle abrasion and monomer. All 3 failure modalities (adhesive, cohesive, and mixed modes) were observed. CONCLUSIONS: Of the materials tested, the most reliable material to bond to milled polymethyl methacrylate was Jet acrylic resin, and the bond strength values were increased substantially when the milled polymethyl methacrylate surface was airborne-particle abraded and monomer was applied.


Assuntos
Colagem Dentária , Polimetil Metacrilato , Óxido de Alumínio , Coroas , Análise do Estresse Dentário , Teste de Materiais , Cimentos de Resina , Resistência ao Cisalhamento , Propriedades de Superfície
20.
J Chin Med Assoc ; 84(2): 125-128, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33306597

RESUMO

Taiwan transitioned to an aged society in 2018. Appropriate dental treatment is important for elderly individuals. Previously, reconstruction of the dentition was thought to help regain chewing function. However, concerns of the elderly population, such as decline in learning ability and saliva secretion, complicate dental reconstruction. Overlooking the special needs of elderly individuals may lead to impaired chewing function, resulting in nutritional imbalances and increased burden on the digestive tract, causing more health disorders. For the elderly population, treatment must be aimed at restoring as much chewing function as possible with minimal changes. Additionally, regular oral hygiene care, proper design of fixed partial dentures, and implant placement greatly reduce the difficulty in adapting to a new prosthesis. These measures allow us to provide better quality of life for elderly individuals.


Assuntos
Assistência Odontológica , Mastigação , Idoso , Força de Mordida , Feminino , Humanos , Masculino , Mastigação/fisiologia , Mecanorreceptores , Qualidade de Vida , Taiwan
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