RESUMEN
The fusion oncoprotein CBFß-SMMHC, expressed in leukemia cases with chromosome 16 inversion, drives leukemia development and maintenance by altering the activity of the transcription factor RUNX1. Here, we demonstrate that CBFß-SMMHC maintains cell viability by neutralizing RUNX1-mediated repression of MYC expression. Upon pharmacologic inhibition of the CBFß-SMMHC/RUNX1 interaction, RUNX1 shows increased binding at three MYC distal enhancers, where it represses MYC expression by mediating the replacement of the SWI/SNF complex component BRG1 with the polycomb-repressive complex component RING1B, leading to apoptosis. Combining the CBFß-SMMHC inhibitor with the BET inhibitor JQ1 eliminates inv(16) leukemia in human cells and a mouse model. Enhancer-interaction analysis indicated that the three enhancers are physically connected with the MYC promoter, and genome-editing analysis demonstrated that they are functionally implicated in deregulation of MYC expression. This study reveals a mechanism whereby CBFß-SMMHC drives leukemia maintenance and suggests that inhibitors targeting chromatin activity may prove effective in inv(16) leukemia therapy.
Asunto(s)
Apoptosis , Cromatina/metabolismo , Proteínas de Fusión Oncogénica/antagonistas & inhibidores , Proteínas Proto-Oncogénicas c-myc/metabolismo , Animales , Apoptosis/efectos de los fármacos , Azepinas/farmacología , Azepinas/uso terapéutico , Bencimidazoles/farmacología , Bencimidazoles/uso terapéutico , Línea Celular Tumoral , Proteínas Cromosómicas no Histona/química , Proteínas Cromosómicas no Histona/metabolismo , Inversión Cromosómica/efectos de los fármacos , Subunidad alfa 2 del Factor de Unión al Sitio Principal/química , Subunidad alfa 2 del Factor de Unión al Sitio Principal/metabolismo , ADN/química , ADN/metabolismo , ADN Helicasas/metabolismo , Modelos Animales de Enfermedad , Humanos , Estimación de Kaplan-Meier , Leucemia Mieloide Aguda/tratamiento farmacológico , Leucemia Mieloide Aguda/mortalidad , Leucemia Mieloide Aguda/patología , Ratones , Ratones Endogámicos C57BL , Proteínas Nucleares/metabolismo , Proteínas de Fusión Oncogénica/metabolismo , Complejo Represivo Polycomb 1/metabolismo , Regiones Promotoras Genéticas , Unión Proteica , Proteínas Proto-Oncogénicas c-myc/genética , Factores de Transcripción/química , Factores de Transcripción/metabolismo , Triazoles/farmacología , Triazoles/uso terapéuticoRESUMEN
Identifying metabolic steps that are specifically required for the survival of cancer cells but are dispensable in normal cells remains a challenge1. Here we report a therapeutic vulnerability in a sugar nucleotide biosynthetic pathway that can be exploited in cancer cells with only a limited impact on normal cells. A systematic examination of conditionally essential metabolic enzymes revealed that UXS1, a Golgi enzyme that converts one sugar nucleotide (UDP-glucuronic acid, UDPGA) to another (UDP-xylose), is essential only in cells that express high levels of the enzyme immediately upstream of it, UGDH. This conditional relationship exists because UXS1 is required to prevent excess accumulation of UDPGA, which is produced by UGDH. UXS1 not only clears away UDPGA but also limits its production through negative feedback on UGDH. Excess UDPGA disrupts Golgi morphology and function, which impedes the trafficking of surface receptors such as EGFR to the plasma membrane and diminishes the signalling capacity of cells. UGDH expression is elevated in several cancers, including lung adenocarcinoma, and is further enhanced during chemoresistant selection. As a result, these cancer cells are selectively dependent on UXS1 for UDPGA detoxification, revealing a potential weakness in tumours with high levels of UGDH.
Asunto(s)
Neoplasias , Uridina Difosfato Ácido Glucurónico , Humanos , Neoplasias/tratamiento farmacológico , Neoplasias/metabolismo , Neoplasias/patología , Transducción de Señal , Uridina Difosfato Ácido Glucurónico/biosíntesis , Uridina Difosfato Ácido Glucurónico/metabolismo , Uridina Difosfato Xilosa/biosíntesis , Uridina Difosfato Xilosa/metabolismo , Adenocarcinoma del Pulmón , Neoplasias PulmonaresRESUMEN
Dietary composition has major effects on physiology. Here, we show that developmental rate, reproduction, and lifespan are altered in C. elegans fed Comamonas DA1877 relative to those fed a standard E. coli OP50 diet. We identify a set of genes that change in expression in response to this diet and use the promoter of one of these (acdh-1) as a dietary sensor. Remarkably, the effects on transcription and development occur even when Comamonas DA1877 is diluted with another diet, suggesting that Comamonas DA1877 generates a signal that is sensed by the nematode. Surprisingly, the developmental effect is independent from TOR and insulin signaling. Rather, Comamonas DA1877 affects cyclic gene expression during molting, likely through the nuclear hormone receptor NHR-23. Altogether, our findings indicate that different bacteria elicit various responses via distinct mechanisms, which has implications for diseases such as obesity and the interactions between the human microbiome and intestinal cells.
Asunto(s)
Proteínas de Caenorhabditis elegans/metabolismo , Caenorhabditis elegans/fisiología , Insulina/metabolismo , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Acil-CoA Deshidrogenasa/metabolismo , Animales , Betaproteobacteria , Caenorhabditis elegans/metabolismo , Dieta , Escherichia coli , Expresión Génica , Longevidad , Muda , Receptores Citoplasmáticos y Nucleares/metabolismo , Inanición , TranscriptomaRESUMEN
Expression profiles are tailored according to dietary input. However, the networks that control dietary responses remain largely uncharacterized. Here, we combine forward and reverse genetic screens to delineate a network of 184 genes that affect the C. elegans dietary response to Comamonas DA1877 bacteria. We find that perturbation of a mitochondrial network composed of enzymes involved in amino acid metabolism and the TCA cycle affects the dietary response. In humans, mutations in the corresponding genes cause inborn diseases of amino acid metabolism, most of which are treated by dietary intervention. We identify several transcription factors (TFs) that mediate the changes in gene expression upon metabolic network perturbations. Altogether, our findings unveil a transcriptional response system that is poised to sense dietary cues and metabolic imbalances, illustrating extensive communication between metabolic networks in the mitochondria and gene regulatory networks in the nucleus.
Asunto(s)
Caenorhabditis elegans/metabolismo , Redes Reguladoras de Genes , Redes y Vías Metabólicas , Acil-CoA Deshidrogenasa/metabolismo , Animales , Betaproteobacteria , Proteínas de Caenorhabditis elegans/metabolismo , Dieta , Escherichia coli , Humanos , Insulina/metabolismo , Errores Innatos del Metabolismo , Mitocondrias/metabolismo , Fosfotransferasas (Aceptor de Grupo Alcohol)/metabolismo , Interferencia de ARN , Transducción de Señal , Factores de Transcripción/metabolismoRESUMEN
Prime editors have a broad range of potential research and clinical applications. However, methods to delineate their genome-wide editing activities have generally relied on indirect genome-wide editing assessments or the computational prediction of near-cognate sequences. Here we describe a genome-wide approach for the identification of potential prime editor off-target sites, which we call PE-tag. This method relies on the attachment or insertion of an amplification tag at sites of prime editor activity to allow their identification. PE-tag enables genome-wide profiling of off-target sites in vitro using extracted genomic DNA, in mammalian cell lines and in the adult mouse liver. PE-tag components can be delivered in a variety of formats for off-target site detection. Our studies are consistent with the high specificity previously described for prime editor systems, but we find that off-target editing rates are influenced by prime editing guide RNA design. PE-tag represents an accessible, rapid and sensitive approach for the genome-wide identification of prime editor activity and the evaluation of prime editor safety.
Asunto(s)
Edición Génica , Genoma , Ratones , Animales , Edición Génica/métodos , ADN/genética , Roturas del ADN de Doble Cadena , Línea Celular , Sistemas CRISPR-Cas , Mamíferos/genéticaRESUMEN
A network of transcription factors (TFs) coordinates transcription with cell cycle events in eukaryotes. Most TFs in the network are phosphorylated by cyclin-dependent kinase (CDK), which limits their activities during the cell cycle. Here, we investigate the physiological consequences of disrupting CDK regulation of the paralogous repressors Yhp1 and Yox1 in yeast. Blocking Yhp1/Yox1 phosphorylation increases their levels and decreases expression of essential cell cycle regulatory genes which, unexpectedly, increases cellular fitness in optimal growth conditions. Using synthetic genetic interaction screens, we find that Yhp1/Yox1 mutations improve the fitness of mutants with mitotic defects, including condensin mutants. Blocking Yhp1/Yox1 phosphorylation simultaneously accelerates the G1/S transition and delays mitotic exit, without decreasing proliferation rate. This mitotic delay partially reverses the chromosome segregation defect of condensin mutants, potentially explaining their increased fitness when combined with Yhp1/Yox1 phosphomutants. These findings reveal how altering expression of cell cycle genes leads to a redistribution of cell cycle timing and confers a fitness advantage to cells.
Asunto(s)
Genes cdc , Proteínas de Saccharomyces cerevisiae , Ciclo Celular/genética , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Quinasas Ciclina-Dependientes/genética , Quinasas Ciclina-Dependientes/metabolismo , Mitosis/genética , Fosforilación , Saccharomyces cerevisiae/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismoRESUMEN
Effectors play important roles in facilitating the infection of plant pathogenic fungi. However, the gene expression regulatory mechanism of effector genes, in particular at the post-transcriptional level, is largely unknown. In this study, we uncovered the post-transcriptional regulation of an effector gene VmSP1 by a miRNA-like RNA (Vm-milR16) facilitating the infection of the apple tree Valsa canker pathogen Valsa mali. Genetic and molecular biological assays indicated that the expression of VmSP1 could be suppressed by Vm-milR16-mediated mRNA cleavage in a sequence-specific manner. During V. mali infection, Vm-milR16 was downregulated, whereas VmSP1 was upregulated, which further indicated the regulation relationship. VmSP1 was further demonstrated to be a secreted protein and could suppress plant immunity. Deletion of VmSP1 did not affect the vegetative growth but significantly reduced the virulence of V. mali. Further study indicated that VmSP1 could interact with the transcription factor MdbHLH189 of apple. Transiently overexpression of MdbHLH189 enhanced host resistance to V. mali by enhancing the expression of apple defense-related genes, together with the increased callose deposition. Silencing of MdbHLH189 compromised host resistance to V. mali. Our findings uncovered the novel epigenetic regulation mechanism of a virulence-associated effector gene mediated by a fungal milRNA at the post-transcriptional level, and the results enriched the understanding of the function and action mechanism of effectors in tree pathogenic fungi.
Asunto(s)
Malus , MicroARNs , MicroARNs/genética , MicroARNs/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Epigénesis Genética , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiología , Malus/metabolismoRESUMEN
BACKGROUND: The mitogen-activated protein kinase (MAPK) cascade is crucial cell signal transduction mechanism that plays an important role in plant growth and development, metabolism, and stress responses. The MAPK cascade includes three protein kinases, MAPK, MAPKK, and MAPKKK. The three protein kinases mediate signaling to downstream response molecules by sequential phosphorylation. The MAPK gene family has been identified and analyzed in many plants, however it has not been investigated in alfalfa. RESULTS: In this study, Medicago sativa MAPK genes (referred to as MsMAPKs) were identified in the tetraploid alfalfa genome. Eighty MsMAPKs were divided into four groups, with eight in group A, 21 in group B, 21 in group C and 30 in group D. Analysis of the basic structures of the MsMAPKs revealed presence of a conserved TXY motif. Groups A, B and C contained a TEY motif, while group D contained a TDY motif. RNA-seq analysis revealed tissue-specificity of two MsMAPKs and tissue-wide expression of 35 MsMAPKs. Further analysis identified MsMAPK members responsive to drought, salt, and cold stress conditions. Two MsMAPKs (MsMAPK70 and MsMAPK75) responds to salt and cold stresses; two MsMAPKs (MsMAPK60 and MsMAPK73) responds to cold and drought stresses; four MsMAPKs (MsMAPK1, MsMAPK33, MsMAPK64 and MsMAPK71) responds to salt and drought stresses; and two MsMAPKs (MsMAPK5 and MsMAPK7) responded to all three stresses. CONCLUSION: This study comprehensively identified and analysed the alfalfa MAPK gene family. Candidate genes related to abiotic stresses were screened by analysing the RNA-seq data. The results provide key information for further analysis of alfalfa MAPK gene functions and improvement of stress tolerance.
Asunto(s)
Medicago sativa , Proteínas Quinasas Activadas por Mitógenos , Estrés Fisiológico , Medicago sativa/genética , Medicago sativa/enzimología , Medicago sativa/fisiología , Proteínas Quinasas Activadas por Mitógenos/genética , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Estrés Fisiológico/genética , Familia de Multigenes , Regulación de la Expresión Génica de las Plantas , Genoma de Planta , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Filogenia , SequíasRESUMEN
Recurrent mutations in the splicing factor U2AF35 are found in several cancers and myelodysplastic syndrome (MDS). How oncogenic U2AF35 mutants promote transformation remains to be determined. Here we derive cell lines transformed by the oncogenic U2AF35(S34F) mutant and identify aberrantly processed pre-mRNAs by deep sequencing. We find that in U2AF35(S34F)-transformed cells the autophagy-related factor 7 (Atg7) pre-mRNA is abnormally processed, which unexpectedly is not due to altered splicing but rather selection of a distal cleavage and polyadenylation (CP) site. This longer Atg7 mRNA is translated inefficiently, leading to decreased ATG7 levels and an autophagy defect that predisposes cells to secondary mutations, resulting in transformation. MDS and acute myeloid leukemia patient samples harboring U2AF35(S34F) have a similar increased use of the ATG7 distal CP site, and previous studies have shown that mice with hematopoietic cells lacking Atg7 develop an MDS-like syndrome. Collectively, our results reveal a basis for U2AF35(S34F) oncogenic activity.
Asunto(s)
Proteína 7 Relacionada con la Autofagia/genética , Transformación Celular Neoplásica/genética , Leucemia Mieloide Aguda/genética , Síndromes Mielodisplásicos/genética , Procesamiento de Término de ARN 3' , Precursores del ARN/genética , ARN Mensajero/genética , Factor de Empalme U2AF/genética , Anciano , Anciano de 80 o más Años , Animales , Autofagia , Proteína 7 Relacionada con la Autofagia/metabolismo , Línea Celular Transformada , Transformación Celular Neoplásica/metabolismo , Transformación Celular Neoplásica/patología , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Leucemia Mieloide Aguda/metabolismo , Leucemia Mieloide Aguda/patología , Masculino , Ratones Endogámicos BALB C , Ratones Desnudos , Persona de Mediana Edad , Mitocondrias/metabolismo , Mitocondrias/patología , Mutación , Síndromes Mielodisplásicos/metabolismo , Síndromes Mielodisplásicos/patología , Poliadenilación , Interferencia de ARN , Precursores del ARN/metabolismo , ARN Mensajero/metabolismo , Factor de Empalme U2AF/metabolismo , Factores de Tiempo , Transfección , Carga TumoralRESUMEN
BACKGROUND: This study examined the knowledge, attitude, and practice (KAP) toward allergic rhinitis (AR) among parents. METHODS: This cross-sectional study enrolled parents of children with AR at Ningbo Hangzhou Bay Hospital between December 2022 and March 2023. A self-administered questionnaire was developed to collect the demographic characteristics, knowledge, attitudes, and practices toward AR. RESULTS: This study included 480 questionnaires, and 78.33% were mothers. The mean knowledge, attitude, and practice scores were 13.49 ± 6.62 (possible range: 0-24), 33.99 ± 3.40 (possible range: 8-40), and 21.52 ± 3.36 (possible range: 5-26), indicating poor knowledge, positive attitudes, and proactive practice. Multivariable logistic regression analysis showed living in urban areas in Ningbo outside Hangzhou Bay New Zone (OR = 4.33, 95%CI: 1.52-12.34, P = 0.006), living in rural areas in Ningbo (OR = 2.15, 95%CI: 1.00-4.59, P = 0.049), being self-employed (OR = 1.99, 95%CI: 1.00-3.95, P = 0.049), monthly income per capita ≥ 20,000 CNY (OR = 1.89, 95%CI: 1.02-3.47, P = 0.042), child with one biological sibling (OR = 0.48, 95%CI: 0.30-0.78, P = 0.003), and ≥ 6 times hospital visits for AR (OR = 2.32, 95%CI: 1.40-3.86, P = 0.001) were independently associated with adequate knowledge. The knowledge (OR = 1.09, 95%CI: 1.05-1.13, P < 0.001) and ≥ 6 times hospital visits for AR (OR = 1.84, 95%CI: 1.06-3.22, P = 0.032) were independently associated with a positive attitude. The knowledge (OR = 1.08, 95%CI: 1.04-1.13, P = 0.001), attitude (OR = 1.41, 95%CI: 1.28-1.55, P < 0.001), monthly income per capita ≥ 20,000 CNY (OR = 3.59, 95%CI: 1.49-8.65, P = 0.004), no previous hospital visit for AR (OR = 0.35, 95%CI: 0.16-0.78, P = 0.003), and ≥ 6 times hospital visits for AR (OR = 0.40, 95%CI: 0.20-0.81, P = 0.011) were independently associated with the practice scores. CONCLUSIONS: The parents of children with AR had poor knowledge but positive attitudes and proactive practice toward AR. This study has identified a need for specific and reliable information initiatives to be introduced as a means of reducing parental concern and ensuring evidence-based strategies for managing children with AR.
Asunto(s)
Conocimientos, Actitudes y Práctica en Salud , Padres , Rinitis Alérgica , Humanos , China , Femenino , Masculino , Estudios Transversales , Adulto , Padres/psicología , Encuestas y Cuestionarios , Persona de Mediana Edad , Niño , Adulto JovenRESUMEN
An extra copy of chromosome 21 causes Down syndrome, the most common genetic disease in humans. The mechanisms contributing to aneuploidy-related pathologies in this syndrome, independent of the identity of the triplicated genes, are not well defined. To characterize aneuploidy-driven phenotypes in trisomy 21 cells, we performed global transcriptome, proteome, and phenotypic analyses of primary human fibroblasts from individuals with Patau (trisomy 13), Edwards (trisomy 18), or Down syndromes. On average, mRNA and protein levels were increased by 1.5-fold in all trisomies, with a subset of proteins enriched for subunits of macromolecular complexes showing signs of posttranscriptional regulation. These results support the lack of evidence for widespread dosage compensation or dysregulation of chromosomal domains in human autosomes. Furthermore, we show that several aneuploidy-associated phenotypes are present in trisomy 21 cells, including lower viability and increased dependency on serine-driven lipid synthesis. Our studies establish a critical role of aneuploidy, independent of triplicated gene identity, in driving cellular defects associated with trisomy 21.
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Aneuploidia , Fibroblastos/patología , Trisomía/genética , Proliferación Celular , Supervivencia Celular , Células Cultivadas , Fibroblastos/metabolismo , Dosificación de Gen/genética , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Humanos , Lípidos/biosíntesis , Sustancias Macromoleculares/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Serina/metabolismo , Transcripción Genética , Regulación hacia ArribaRESUMEN
Vertebrate mammals express a protein called Ki-67 which is most widely known as a clinically useful marker of highly proliferative cells. Previous studies of human cells indicated that acute depletion of Ki-67 can elicit a delay at the G1/S boundary of the cell cycle, dependent on induction of the checkpoint protein p21. Consistent with those observations, we show here that acute Ki-67 depletion causes hallmarks of DNA damage, and the damage occurs even in the absence of checkpoint signaling. This damage is not observed in cells traversing S phase but is instead robustly detected in mitotic cells. The C-terminal chromatin-binding domain of Ki-67 is necessary and sufficient to protect cells from this damage. We also observe synergistic effects when Ki-67 and p53 are simultaneously depleted, resulting in increased levels of chromosome bridges at anaphase, followed by the appearance of micronuclei. Therefore, these studies identify the C terminus of Ki-67 as an important module for genome stability.
Asunto(s)
Cromatina/metabolismo , Cromosomas Humanos , Antígeno Ki-67/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Anafase , Sitios de Unión , Línea Celular , Daño del ADN , Inestabilidad Genómica , Humanos , Antígeno Ki-67/genética , Mitosis , Dominios Proteicos , Proteína p53 Supresora de Tumor/genética , Proteína 1 de Unión al Supresor Tumoral P53/genética , Proteína 1 de Unión al Supresor Tumoral P53/metabolismoRESUMEN
Germ cells possess the Piwi-interacting RNA pathway to repress transposable elements and maintain genome stability across generations. Transposable element mobilization in somatic cells does not affect future generations, but nonetheless can lead to pathological outcomes in host tissues. We show here that loss of function of the conserved zinc-finger transcription factor Hinfp causes dysregulation of many host genes and derepression of most transposable elements. There is also substantial DNA damage in somatic tissues of Drosophila after loss of Hinfp. Interference of transposable element mobilization by reverse-transcriptase inhibitors can suppress some of the DNA damage phenotypes. The key cell-autonomous target of Hinfp in this process is Histone1, which encodes linker histones essential for higher-order chromatin assembly. Transgenic expression of Hinfp or Histone1, but not Histone4 of core nucleosome, is sufficient to rescue the defects in repressing transposable elements and host genes. Loss of Hinfp enhances Ras-induced tissue growth and aging-related phenotypes. Therefore, Hinfp is a physiological regulator of Histone1-dependent silencing of most transposable elements, as well as many host genes, and serves as a venue for studying genome instability, cancer progression, neurodegeneration, and aging.
Asunto(s)
Elementos Transponibles de ADN/genética , Drosophila melanogaster/genética , Inestabilidad Genómica/genética , Proteínas Represoras/metabolismo , Factores de Transcripción/metabolismo , Animales , Cromatina/metabolismo , Ensamble y Desensamble de Cromatina/genética , Regulación del Desarrollo de la Expresión Génica/genética , Histonas/genética , Histonas/metabolismo , ARN Interferente Pequeño/genéticaRESUMEN
OBJECTIVES: To investigate the function of circMyt1l/rno-let-7d-5p/BDNF in the white matter damage of premature rats. METHODS: Bioinformatic analysis was used to analyze the differential expression of circMyt1l and its interacting miRNAs and mRNAs in rats with periventricular white matter damage. Rats at postnatal day 3 had their right common carotid artery permanently ligated, and were then exposed for 2â¯h to 6â¯% O2, or sham surgery and exposure to normal O2 levels (sham). CircMyt1l and rno-let-7d-5p expression was detected and BDNF protein levels were analyzed at 24, 48, and 72â¯h post hypoxia-ischemia. RESULTS: Bioinformatic analysis suggested that circMyt1l, rno-let-7d-5p and BDNF interact. CircMyt1l expression decreased significantly relative to the sham-operated rats (p<0.01) in an exposure time-dependent manner. Contrastingly, rno-let-7d-5p increased significantly relative to the sham-operated rats (p<0.01) in an exposure time dependent manner. BDNF protein levels decreased significantly relative to the sham-operated rats (p<0.05) in an exposure time dependent manner. CONCLUSIONS: The expression levels of circMyt1l/rno-let-7d-5p/BDNF are interrelated in periventricular white matter damage. Decreased circMyt1l expression of promoted the effect of rno-let-7d-5p and decreased the level of its target, BDNF.
Asunto(s)
MicroARNs , Sustancia Blanca , Ratas , Animales , Animales Recién Nacidos , Factor Neurotrófico Derivado del Encéfalo/metabolismo , Sustancia Blanca/diagnóstico por imagen , Sustancia Blanca/metabolismo , MicroARNs/metabolismoRESUMEN
Abnormalities in the zona pellucida (ZP) adversely affect oocyte maturation, embryo development and pregnancy outcomes. However, the assessment of severity is challenging. To evaluate the effects of different degrees of ZP abnormalities on embryo development and clinical outcomes, in total, 590 retrieval cycles were scored and divided into four categories (control, mild, moderate and severe) based on three parameters: perivitelline space, percentage of immature oocytes and percentage of oocytes with abnormal morphology. As the severity of abnormal ZP increased, both the number of retrieved oocytes and mature oocytes decreased. The fertilization rate did not differ significantly among groups. The rates of embryo cleavage and day-3 high-quality embryos in the mild group and the moderate group did not vary significantly between the two groups but were significantly higher than those in the severe group. The blastulation rates of the abnormal ZP groups were similar; however, they were lower than those of the control group. Moreover, the cycle cancellation rate of the severe abnormal ZP group was as high as 66.20%, which was significantly higher than that of the other three groups. Although the rates of cumulative clinical pregnancy and live births were lower than those in the control group, they were comparable among the abnormal ZP groups. There were no differences in the neonatal outcomes of the different groups. Together, ZP abnormalities show various degrees of severity, and in all patients regardless of the degree of ZP abnormalities who achieve available embryos, there will be an opportunity to eventually give birth.
Asunto(s)
Fertilización In Vitro , Zona Pelúcida , Embarazo , Femenino , Recién Nacido , Humanos , Oocitos , Resultado del Embarazo , Inyecciones de Esperma IntracitoplasmáticasRESUMEN
Jasminum mesnyi Hance is an important medicinal and ornamental plant. This species is native to South Central China and Vietnam and grows primarily in the subtropical biomes. In June 2022, 17 Colletotrichum strains were isolated from leaf tip blight on foliage of J. mesnyi in Nanjing, Jiangsu, China. Based on morphological characteristics and multilocus phylogenetic analyses of six genomic loci (ITS, CAL, ACT, TUB2, CHS-1, and GAPDH), a new species, namely, C. nanjingense, and a known species, namely, C. gloeosporioides s.s., were described and reported. Pathogenicity tests revealed that both species were pathogens causing leaf tip blight on J. mesnyi. The results provided necessary information for disease control and enhanced our understanding of the diversity of Colletotrichum species in China.
Asunto(s)
Colletotrichum , Jasminum , Jasminum/genética , Filogenia , Enfermedades de las Plantas , ADN de Hongos/genética , China , Hojas de la PlantaRESUMEN
Methamphetamine (METH) is a highly addictive psychostimulant and one of the most widely abused drugs worldwide. The continuous use of METH eventually leads to neurotoxicity and drug addiction. Studies have shown that neurotoxicity is strongly associated with METH-induced neuroinflammation, and microglia are the key drivers of neuroinflammation. Triggering receptor expressed on myeloid cells 2 (TREM2) is reported to play a key role in activation of microglia and neuroinflammation. Yet, the molecular mechanisms by which METH causes neuroinflammation and neurotoxicity remain elusive. In the current study, we investigated the role of TREM2 in neuroinflammation induced by METH in BV2 cells and the wild-type (WT) C57BL/6J mice, CX3CR1GFP/+ transgenic mice, and TREM2 knockout (KO) mice. Postmortem samples from the frontal cortex of humans with a history of METH use were also analyzed to determine the levels of TREM2, TLR4, IBA1, and IL-1ß. The expression levels of TREM2, TLR4, IBA1, IL-1ß, iNOS, and Arg-1 were then assessed in the BV2 cells and frontal cortex of mice and human METH users. Results revealed that the expression levels of TREM2, TLR4, IBA1, and IL-1ß were significantly elevated in METH-using individuals and BV2 cells. Microglia were clearly activated in the frontal cortex of WT C57BL/6 mice and CX3CR1GFP/+ transgenic mice, and the protein levels of IBA1, TREM2, TLR4, and IL-1ß were elevated in the METH-induced mouse models. Moreover, TREM2-KO mice showed further increased microglial activation, neuroinflammation, and excitotoxicity induced by METH. Thus, these findings suggest that TREM2 may be a target for regulating METH-induced neuroinflammation.
Asunto(s)
Metanfetamina , Humanos , Animales , Ratones , Metanfetamina/toxicidad , Microglía/metabolismo , Enfermedades Neuroinflamatorias , Receptor Toll-Like 4/metabolismo , Ratones Endogámicos C57BL , Ratones Transgénicos , Células Mieloides/metabolismoRESUMEN
Watermelon is one of the most important edible plants worldwide. Owing to its special cultivation conditions, watermelon is exposed to many biological and abiotic stresses during its development. Lectin receptor-like kinases (LecRLKs) are plant-specific membrane proteins that play important roles in sensing and responding to environmental stimuli. Although the LecRLK gene family has been identified in a variety of plants, a comprehensive analysis has not yet been undertaken in watermelon. In this study, 61 putative LecRLK genes were identified in watermelon, consisting of 36 G-type, 24 L-type, and 1 C-type LecRLK genes. They were distributed in clusters on chromosomes, and members from the same subfamily were mostly clustered together. The analysis of the phylogenetic tree and conserved motif indicated that there were obvious differences among three ClaLecRLK subfamilies, and there was also rich diversity in the C-terminal within subfamilies. A collinear analysis revealed that the evolution of the ClaLecRLK gene family in different Cucurbitaceae crops was asynchronous. Furthermore, the analysis of the ClaLecRLK protein structure showed that not all proteins contained signal peptides and a single transmembrane domain. A subcellular localization assay confirmed that the number and position of transmembrane domains did not affect ClaLecRLK protein localization in cells. Transcriptome data revealed distinct expression patterns of LecRLK genes of watermelon in various tissues, and their responses to different fungi infection were also significantly different. Finally, the potential binding sites of the ClaLecRLK genes targeted by miRNA were predicted. This study enhances the understanding of the characteristics and functions of the LecRLK gene family in watermelon and opens up the possibility of exploring the roles that LecRLK genes may play in the life cycle of Cucurbitaceae plants.
Asunto(s)
Citrullus , Regulación de la Expresión Génica de las Plantas , Filogenia , Proteínas de Plantas , Citrullus/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Perfilación de la Expresión Génica , Genoma de Planta , Estudio de Asociación del Genoma Completo , Familia de Multigenes , Cromosomas de las Plantas/genéticaRESUMEN
Metabolic engineering enables oilseed crops to be more competitive by having more attractive properties for oleochemical industrial applications. The aim of this study was to increase the erucic acid level and to produce wax ester (WE) in seed oil by genetic transformation to enhance the industrial applications of B. carinata. Six transgenic lines for high erucic acid and fifteen transgenic lines for wax esters were obtained. The integration of the target genes for high erucic acid (BnFAE1 and LdPLAAT) and for WEs (ScWS and ScFAR) in the genome of B. carinata cv. 'Derash' was confirmed by PCR analysis. The qRT-PCR results showed overexpression of BnFAE1 and LdPLAAT and downregulation of RNAi-BcFAD2 in the seeds of the transgenic lines. The fatty acid profile and WE content and profile in the seed oil of the transgenic lines and wild type grown in biotron were analyzed using gas chromatography and nanoelectrospray coupled with tandem mass spectrometry. A significant increase in erucic acid was observed in some transgenic lines ranging from 19% to 29% in relation to the wild type, with a level of erucic acid reaching up to 52.7%. Likewise, the transgenic lines harboring ScFAR and ScWS genes produced up to 25% WE content, and the most abundant WE species were 22:1/20:1 and 22:1/22:1. This study demonstrated that metabolic engineering is an effective biotechnological approach for developing B. carinata into an industrial crop.