RESUMEN
Blood transcriptomics has emerged as a vital tool for tracking the immune system and supporting disease diagnosis, prognosis, treatment, and research. The present study was conducted to analyze the gene expression profile and potential biomarker candidates using the whole blood of mandarin fish (Siniperca chuatsi) infected with LPS or poly (I:C) at 0 h, 3 h, 6 h, and 12 h. Our data suggest that 310 shared differentially expressed genes (DEGs) were identified among each comparison group after LPS infection, and 137 shared DEGs were identified after poly (I:C) infection. A total of 62 shared DEGs were differentially expressed in all compared groups after LPS or poly (I:C) infection. Pathways analysis for DEGs in all different compared groups showed that cytokine-cytokine receptor interaction was the most enrichment pathway. The expression levels of genes C-X-C chemokine receptor type 2-like (cxcr2), chemokine (C-C motif) receptor 9a (ccr9a), chemokine (C-C motif) receptor 9b (ccr9b), chemokine (C-X-C motif) receptor 4b (cxcr4b), and interleukin 10 receptor alpha (il10ra) were significantly different in all compared groups and most enriched in cytokine-cytokine receptor interaction pathway. The protein-protein interactions analysis among all shared DEGs showed that cxcr4 was the hub gene with the highest degree. The biomarker candidates discovered in this study may, following validation, prove effective as diagnostic tools in monitoring mandarin fish diseases.
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The coronavirus disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spread worldwide for more than 3 years. Although the hospitalization rate and mortality have decreased dramatically due to wide vaccination effort and improved treatment options, the disease is still a global health issue due to constant viral mutations, causing negative impact on social and economic activities. In addition, long COVID and complications arising from COVID-19 weeks after infection have become a concern for public health experts. Therefore, better treatments for COVID-19 are still needed. Herein, we describe a class of macrocyclic peptidomimetic compounds that are potent inhibitors of SARS-Cov-2 3CL protease (3CLpro). Significantly, some of the compounds showed a higher stability against human liver microsomes (HLM t1/2 > 180 min) and may be suitable for oral administration without the need for a pharmacokinetic (PK) boosting agent such as ritonavir.
Asunto(s)
Antivirales , Proteasas 3C de Coronavirus , Compuestos Macrocíclicos , SARS-CoV-2 , Proteasas 3C de Coronavirus/antagonistas & inhibidores , Proteasas 3C de Coronavirus/metabolismo , Humanos , SARS-CoV-2/efectos de los fármacos , Compuestos Macrocíclicos/química , Compuestos Macrocíclicos/farmacología , Compuestos Macrocíclicos/síntesis química , Compuestos Macrocíclicos/farmacocinética , Antivirales/farmacología , Antivirales/química , Antivirales/síntesis química , Antivirales/farmacocinética , Microsomas Hepáticos/metabolismo , Peptidomiméticos/farmacología , Peptidomiméticos/química , Peptidomiméticos/síntesis química , Descubrimiento de Drogas , Tratamiento Farmacológico de COVID-19 , Inhibidores de Proteasas/farmacología , Inhibidores de Proteasas/química , Inhibidores de Proteasas/síntesis química , Inhibidores de Proteasas/farmacocinética , Relación Estructura-ActividadRESUMEN
Carotenoid cleavage oxygenases can cleave carotenoids into a range of biologically important products. Carotenoid isomerooxygenase (NinaB) and ß, ß-carotene 15, 15'-monooxygenase (BCO1) are two important oxygenases. In order to understand the roles that both oxygenases exert in crustaceans, we first investigated NinaB-like (EsNinaBl) and BCO1-like (EsBCO1l) within the genome of Chinese mitten crab (Eriocheir sinensis). Their functions were then deciphered through an analysis of their expression patterns, an in vitro ß-carotene degradation assay, and RNA interference. The results showed that both EsNinaBl and EsBCO1l contain an RPE65 domain and exhibit high levels of expression in the hepatopancreas. During the molting stage, EsNinaBl exhibited significant upregulation in stage C, whereas EsBCO1l showed significantly higher expression levels at stage AB. Moreover, dietary supplementation with ß-carotene resulted in a notable increase in the expression of EsNinaBl and EsBCO1l in the hepatopancreas. Further functional assays showed that the EsNinaBl expressed in E. coli underwent significant changes in its color, from orange to light; in addition, its ß-carotene cleavage was higher than that of EsBCO1l. After the knockdown of EsNinaBl or EsBCO1l in juvenile E. sinensis, the expression levels of both genes were significantly decreased in the hepatopancreas, accompanied by a notable increase in the redness (a*) values. Furthermore, a significant increase in the ß-carotene content was observed in the hepatopancreas when EsNinaBl-mRNA was suppressed, which suggests that EsNinaBl plays an important role in carotenoid cleavage, specifically ß-carotene. In conclusion, our findings suggest that EsNinaBl and EsBCO1l may exhibit functional co-expression and play a crucial role in carotenoid cleavage in crabs.
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Braquiuros , Hepatopáncreas , beta Caroteno , beta-Caroteno 15,15'-Monooxigenasa , Animales , beta Caroteno/metabolismo , Braquiuros/metabolismo , Braquiuros/genética , beta-Caroteno 15,15'-Monooxigenasa/metabolismo , beta-Caroteno 15,15'-Monooxigenasa/genética , Hepatopáncreas/metabolismo , Muda/genética , Oxigenasas/metabolismo , Oxigenasas/genética , Filogenia , Proteínas de Artrópodos/genética , Proteínas de Artrópodos/metabolismoRESUMEN
The mandarin fish (Siniperca chuatsi) DNA methyltransferase gene 1 (dnmt1) was highly expressed in the mesonephros, head kidney and gonad, whereas dnmt2 was expressed in most tissues. dnmt3a was highly expressed in the brain and spleen, but dnmt3b was mainly expressed in the brain and head kidney. The genes dnmt1 and dnmt2 were highly expressed in the early stages of embryonic development, and dnmt3a and dnmt3b were expressed later. These genes also showed certain changes after artificial diet acclimation, salinity adaptation and immune stress.
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Perciformes , Animales , ADN , Perfilación de la Expresión Génica , Metiltransferasas , Perciformes/genética , TranscriptomaRESUMEN
Oreochromis niloticus RAS-association domain family 4 (rassf4) was specifically expressed in the ovaries. Immunohistochemistry results showed that Rassf4 was located in follicular cells. The methylation percentages of the promoter region (-734/-1012) of rassf4 in the ovaries and testes were 13.28% and 92.85%, respectively. Deleting the fragment of -734 to -1012 sites significantly reduced the basal activity of the rassf4 promoter to 62.33%, which indicated that this region was important for the transcription of the rassf4 gene. This study lays the foundation for further research on the function of fish rassf4.
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Cíclidos , Animales , Cíclidos/genética , Metilación de ADN , Femenino , Masculino , Ovario , Regiones Promotoras Genéticas , TestículoRESUMEN
Mandarin fish (Siniperca chuatsi) is an important economic fish in China. Viral and bacterial diseases seriously affect the artificial culture of S. chuatsi. As a carnivorous fish, artificial feed domestication is also an important means to improve the scale of S. chuatsi culture. Therefore, the study of immunology and digestive physiology is very important to the industrial development of S. chuatsi. In this work, we analyzed the expression and function of the S. chuatsi leukocyte cell-derived chemotaxin 2 (Sc-lect2) gene on a basis of next generation, single-molecule long-read sequencing. Sc-lect2 was mainly expressed in the liver but barely expressed in the gill, skin, muscle, kidney, head kidney, brain, stomach, and intestine. When the fish were infected with infectious spleen and kidney necrosis virus and challenged with lipopolysaccharide and polyinosinic-polycytidylic acid, Sc-lect2 expression significantly increased by about 40, 17, and 7-fold, respectively, compared with unstimulated samples. We also found that Sc-lect2 increases by approximately 8-fold after the fish are fed an artificial diet. These results show that mandarin fish liver can not only digest food but also express specific immune genes. Changes in the diet can cause the differential expression of Sc-lect2 genes. Four Sc-lect2 interaction genes were differentially expressed in the skin or blood. Interestingly, miR-145-3p could inhibit Sc-lect2 gene expression by targeting its coding sequence region. One CpG island in the promoter region showed a high level of methylation, suggesting that high methylation does not affect Sc-lect2 gene expression in the liver.
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Enfermedades de los Peces/inmunología , Regulación de la Expresión Génica/inmunología , Inmunidad Innata/genética , Péptidos y Proteínas de Señalización Intercelular/genética , Péptidos y Proteínas de Señalización Intercelular/inmunología , Perciformes/genética , Perciformes/inmunología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Proteínas de Peces/química , Proteínas de Peces/genética , Proteínas de Peces/inmunología , Perfilación de la Expresión Génica/veterinaria , Péptidos y Proteínas de Señalización Intercelular/química , Filogenia , Estructura Terciaria de Proteína , Alineación de Secuencia/veterinariaRESUMEN
The viral hemorrhage disease caused by grass carp reovirus (GCRV) is a serious contagious disease of grass carp that mainly infects fingerlings and yearlings. Epidemiological studies have shown that GCRV genotype II is currently the prominent genotype. However, little is known about the histopathological characteristics, virus distribution, and expression of immunity-related genes in grass carp infected by GCRV genotype II. In this study, we found that grass carp infected by GCRV genotype II lost appetite, swam alone, and rolled, and their fins, eyes, operculum, oral cavity, abdomen, intestine, and muscles showed pronounced punctate hemorrhage. Congestion, swelling, deformation, thinning of membranes, dilatation and darkened color of nucleoli, cathepsis, erythrocyte infiltration, and vacuole formation were observed in some infected tissues. A qRT-PCR test showed that the 11 genome segments of GCRV had similar expression patterns in different tissues. The S8 segment, with unknown function and no homologous sequences, had the highest expression level, while the most conserved segment, L2, had the lowest expression level. GCRV particles were distributed in different tissues, especially in the intestine. In the infected intestine, the expression of various receptors and adaptor molecules was modulated at different levels. Pro-inflammatory cytokine interleukin-1ß (IL-1ß) expression was 2160.9 times higher than that in the control group. The upregulation of immunity-related genes activated the antiviral immunity pathways. Therefore, the intestine might play a dual role in mediating GCRV infection and the antiviral immune response. This study provides detailed information about the pathogenicity of GCRV and expression of immunity-related genes, laying the foundation for further research on virus control and treatment.
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Carpas/virología , Enfermedades de los Peces/genética , Proteínas de Peces/genética , Infecciones por Reoviridae/veterinaria , Reoviridae/fisiología , Animales , Carpas/genética , Carpas/inmunología , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/patología , Enfermedades de los Peces/virología , Proteínas de Peces/inmunología , Genoma Viral , Genotipo , Interleucina-1beta/genética , Interleucina-1beta/inmunología , Reoviridae/genética , Infecciones por Reoviridae/genética , Infecciones por Reoviridae/inmunología , Infecciones por Reoviridae/virologíaRESUMEN
Animal pigmentation primarily depends on the presence and mixing ratio of chromatophores, functioning in animal survival and communication. For the benthic and carnivorous Siniperca chuatsi, pigmentation pattern is key to concealment and predation. In this study, the formation, distribution, and main pattern of chromatophores were observed in the embryos, larvae, skins, and visceral tissues from S. chuatsi. Melanophores were firstly visualized in the yolk sac at segmentation stage, and then they were migrated to the whole body and further clustered into the black stripes, bands, and patches. In adult S. chuatsi, the head, black band, and body side skins mainly contained melanophores, showing as deep or light black. The abdomen skin mainly contained iridophores, showing as silvery. In the eye, the pigment layers were located in the epithelial layers of iris and retina and shown as black. Then, the pigmentation-related gene, tyrosinase gene from S. chuatsi (Sc-tyr) was analyzed by bioinformatics and quantitative methods. The Sc-tyr gene encoded a protein with 540 amino acids (Sc-TYR). The Sc-TYR contained two copper ion binding sites, which were coordinated by six conserved histidines (H182, H205, H214, H366, H370, H393) and necessary for catalytic activity. The Sc-TYR was well conserved compared with TYR of various species with higher degree of sequence similarity with other fishes (77.6-98.3%). The qRT-PCR test showed that the Sc-tyr mRNA reached the peak value at segmentation stage in the embryo development, the black skins displayed a higher expression level than that in silvery skin, and the eye had the highest expression level compared with other tissues. Further research on enzyme activity showed that the expression patterns of tyrosinase activity were similar to that of the Sc-tyr mRNA. Comparing with the results of molecular and phenotype, it was found that the temporal and spatial distributions of tyrosinase corresponded well with changes in pigmentation patterns and the intensity of skin melanization. This study initially explored the pigmentation formation and tyrosinase expression, which served as a foundation for further insight into the genetics mechanism of body color formation in S. chuatsi.
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Cromatóforos/fisiología , Peces/fisiología , Monofenol Monooxigenasa/biosíntesis , Pigmentación/fisiología , Conducta Predatoria/fisiología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Biología Computacional , Peces/clasificación , Peces/embriología , Peces/genética , Secciones por Congelación , Riñón/anatomía & histología , Larva/anatomía & histología , Melanóforos/fisiología , Melanóforos/ultraestructura , Conformación Molecular , Monofenol Monooxigenasa/química , Monofenol Monooxigenasa/genética , Filogenia , Epitelio Pigmentado Ocular/anatomía & histología , Epitelio Pigmentado Ocular/fisiología , Conformación Proteica , Alineación de Secuencia , Piel/anatomía & histología , Piel/enzimología , Bazo/anatomía & histologíaRESUMEN
Methyltransferase-like 8 (mettl8) is a protein-coding gene that may demonstrate nucleic acid or protein methyltransferase activity. Although several members of the METTL protein family have been reported, the expression and function of this family are still poorly understood, especially in fish. Medaka (Oryzias latipes) is an important model organism with relatively complete genome information, and more and more genetic toolkits are available for this fish. The popularity of medaka among developmental biologists has led to important insights into vertebrate development. Here, we report the DNA sequence and expression of mettl8 in medaka. The full-length cDNA of medaka mettl8 is 1266 bp, and its predicted open reading frame codes for a protein with 393 amino acids. The predicted molecular mass was 45.8 kDa, and the theoretical isoelectric point was 8.61. It had a conserved methyltransferase domain in METTL8 proteins. Homology analysis revealed that medaka METTL8 clustered in close proximity with the METTL8 of Austrofundulus limnaeus and Nothobranchius furzeri within the Cyprinodontiformes branch, and the protein structure of METTL8 was highly conserved. During embryogenesis, the mettl8 transcript was highly expressed in early stages, while it persisted at a detectable level until the larvae stage. In adult fish, the RT-PCR result indicated that mettl8 mRNA was expressed in the brain, eye, skin, liver, intestine, ovary, and testis. Slice in situ hybridization analysis showed that mettl8 was highly expressed in the eye, intestine, ovary, and testis. The expression and distribution of mettl8 during embryogenesis were also demonstrated by whole mount in situ hybridization. The results indicated that the mettl8 is expressed significantly in the eye, somite, and otic vesicles. Immunofluorescence and Western blot analyses showed that METTL8 protein was present in both the nuclei and cytoplasm. This study lays a foundation for further research on the function of fish mettl8.
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Proteínas de Peces/genética , Metiltransferasas/genética , Oryzias/genética , Animales , Secuencia de Bases , Encéfalo/metabolismo , ADN Complementario/genética , Ojo/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Mucosa Intestinal/metabolismo , Hígado/metabolismo , Masculino , Ovario/metabolismo , Filogenia , ARN Mensajero/metabolismo , Piel/metabolismo , Testículo/metabolismoRESUMEN
The bromodomain and extra-terminal (BET) family of proteins, consisting of the bromodomains containing protein 2 (BRD2), BRD3, BRD4, and the testis-specific BRDT, are key epigenetic regulators of gene transcription and has emerged as an attractive target for anticancer therapy. Herein, we describe the discovery of a novel potent BET bromodomain inhibitor, using a systematic structure-based approach focused on improving potency, metabolic stability, and permeability. The optimized dimethylisoxazole aryl-benzimidazole inhibitor exhibited high potency towards BRD4 and related BET proteins in biochemical and cell-based assays and inhibited tumor growth in two proof-of-concept preclinical animal models.
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Bencimidazoles/farmacología , Descubrimiento de Drogas , Isoxazoles/farmacología , Mieloma Múltiple/tratamiento farmacológico , Factores de Transcripción/antagonistas & inhibidores , Administración Oral , Animales , Bencimidazoles/química , Bencimidazoles/metabolismo , Disponibilidad Biológica , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Humanos , Isoxazoles/administración & dosificación , Isoxazoles/química , Isoxazoles/metabolismo , Ratones , Estructura Molecular , Mieloma Múltiple/metabolismo , Mieloma Múltiple/patología , Neoplasias Experimentales/tratamiento farmacológico , Neoplasias Experimentales/metabolismo , Neoplasias Experimentales/patología , Dominios Proteicos/efectos de los fármacos , Relación Estructura-Actividad , Factores de Transcripción/metabolismoRESUMEN
OBJECTIVE: This study aimed to clarify the association between an increased spot urine albumin-to-creatinine ratio (UACR) and the risk of stroke. METHODS: We performed a systematic review and meta-analysis of cohort studies, case-control studies, and ancillary data randomized controlled trials (RCTs), which were treated as cohorts in this study, and estimated the association between albuminuria, as measured with the UACR, and the risk of stroke. We performed a comprehensive search of PubMed, Embase, and the Cochrane Library and conducted a systematic review and cumulative meta-analysis of cohort studies with a cross-sectional with prospective design in which stroke incidence was reported and the baseline UACR was measured. Ancillary data from RCTs were also included as part of the cohort study. We studied the characteristics of the participants, quality scores and risk ratios (RR, with confidence intervals, CI) of stroke associated with normal and high UACRs, and we synthesized the data via a meta-analysis. RESULTS: Twelve eligible studies including a total of 32,888 participants and 3,944 cases of stroke were identified. A high UACR (>30 mg/mmol) increased the risk of stroke by 1.67 times (RR: 1.67, 95% CI: 1.49-1.86, P<0.001 I2â¯=â¯26%). The results were not different between Asian and non-Asian patients (RR: 1.64, 95% CI: 1.41-1.91, P<0.001, I2â¯=â¯23% compared with RR: 1.67, 95% CI: 1.50-1.85, P<0. 00, I2â¯=â¯39%) or between subgroups classified by old age (RR: 1.61, 95% CI: 1.39-1.88, P<0.001, I2â¯=â¯34% compared with RR: 1.68, 95% CI: 1.52-1.87, P<0.001, I2â¯=â¯13%). A sensitivity analysis did not significantly change the results. CONCLUSION: The incidence of stroke increased significantly in the high UACR group compared with the normal UACR group. The UACR could be a clinical addition for the early indication of high-risk stroke patients.
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Albuminuria/epidemiología , Biomarcadores/orina , Creatinina/orina , Accidente Cerebrovascular/epidemiología , Anciano , Albuminuria/diagnóstico , Albuminuria/orina , Diagnóstico Precoz , Femenino , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Factores de Riesgo , Accidente Cerebrovascular/diagnóstico , Accidente Cerebrovascular/orina , UrinálisisRESUMEN
Hepcidin is a liver-derived peptide hormone that is related to iron balance and immunity in humans. However, its function in Siniperca chuatsi has not been well elucidated. In this study, we analyzed the expression and function of the S. chuatsi hepcidin (Sc-hep) gene. Sc-hep was specifically expressed in the liver and appeared to be one of the most highly expressed genes in the liver. After spleen and kidney necrosis virus (ISKNV) infection and lipopolysaccharide (LPS) and polyinosinic-polycytidylic acid (Poly I:C) stimulation, the expression of Sc-hep in the liver increased by approximately 110, 6500, and 225 times, respectively. After ferrous sulfate (FS) injection, the expression of Sc-hep in the liver increased approximately 520-fold. We found that miR-19c-5p could inhibit Sc-hep expression. Five CpG dinucleotides distributed in the promoter region showed no differential methylation between the liver and the stomach, both presenting high methylation rates. After FS or LPS injection, the expression of three iron balance-related genes (FPN1, TFR1, and FTN) and five immune-related cytokine genes (IL-1ß, IL8, TNF-α, TLR22, and SOCS3) significantly changed. These results indicate that Sc-hep participates in the regulation of iron balance and plays an important role in the immune system. Sc-hep increased approximately 52-fold when mandarin fish were domesticated with artificial diets. Sc-hep might be used as a real-time biomarker of mandarin fish liver because its expression markedly varies under different physiological conditions.
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Enfermedades de los Peces/genética , Proteínas de Peces/genética , Peces/genética , Hepcidinas/genética , Animales , Metilación de ADN , Infecciones por Virus ADN/genética , Infecciones por Virus ADN/inmunología , Infecciones por Virus ADN/veterinaria , Enfermedades de los Peces/inmunología , Enfermedades de los Peces/virología , Proteínas de Peces/inmunología , Peces/inmunología , Peces/virología , Regulación de la Expresión Génica , Hepcidinas/inmunología , Inmunidad , Iridoviridae/inmunología , Lipopolisacáridos/inmunología , MicroARNs/genética , MicroARNs/inmunología , FilogeniaRESUMEN
Interferon regulatory factors (IRFs) are transcription factors of the interferon (IFN)-inducible signaling pathway essential for host immunity against antimicrobial infection by virus and bacteria. Interferon regulatory factor 3 (IRF3) regulates the expression of IFNs and IFN-stimulated genes by binding to the IFN stimulatory response element (ISRE). In this study, we analyze the thymus transcriptome of the mandarin fish Siniperca chuatsi and report the functional analysis of Irf3 from the thymus as an emerging model of antiviral approaches. The predicted S. chuatsi IRF3 (Sc-Irf3) protein has 465 amino acid residues and evolutionarily conserved domains and is clustered in the IRF3 subfamily on a phylogenetic tree. Sc-Irf3 upon transgenic expression was mainly found in the cytoplasm through Western blot analysis and microscopy, but it translocated to the nucleus after polyinosinic:polycytidylic acid (ploly I:C) treatment. Endogenous Sc-irf3 RNA expression was detected in all eight adult organs examined. Importantly, Sc-irf3 RNA expression was significantly upregulated by ploly(I:C) treatment in the adult organs. Concurrently, reporter assays revealed that Sc-Irf3 increased the transcriptional activity of the ifnß promoter, a minimal ISRE-containing promoter, and ifn promoter of mandarin fish. Therefore, Sc-Irf3 plays a major role in the IFN immune defense system against virus infection.
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Regulación de la Expresión Génica/fisiología , Factor 3 Regulador del Interferón/metabolismo , Interferones/metabolismo , Perciformes/fisiología , Timo/metabolismo , Animales , Factor 3 Regulador del Interferón/genética , Interferones/genéticaRESUMEN
Manganese (Mn) is required in various human physiological processes. Excessive Mn exposure causes manganism, a progressive neurodegenerative disorder similar to idiopathic Parkinson's disease (IPD). However, the detailed mechanism of Mn-induced neurotoxicity is not yet fully understood. MicroRNAs (miRNAs) play important roles in gene expression regulation, and miRNA expression profile provides additional biological and prognostic information of diseases. In our study, RNA sequencing was performed to profile miRNAs in the SH-SY5Y cells following MnCl2 treatment. Expressions of 73 miRNAs were altered following excessive Mn treatment. Furthermore, has-miR-4306 was identified to target 3'UTR of ATP13A2 (PARK9) directly. Inhibition of has-miR-4306 efficiently restored Mn-induced cytotoxicity. Thus, for the first time, we revealed the miRNA effects of Mn ions to neuron cells, highlighted the involvement of miRNA regulation in neurodegeneration caused by Mn exposure, and provided a potential application of miRNAs in future therapeutic intervention.
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Cloruros/toxicidad , Regulación de la Expresión Génica , MicroARNs/genética , Neuronas/efectos de los fármacos , ATPasas de Translocación de Protón/genética , Secuencia de Bases , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Perfilación de la Expresión Génica , Humanos , Compuestos de Manganeso , MicroARNs/metabolismo , Modelos Biológicos , Neuronas/metabolismo , Neuronas/patología , Enfermedad de Parkinson/genética , Enfermedad de Parkinson/metabolismo , Enfermedad de Parkinson/patología , ATPasas de Translocación de Protón/antagonistas & inhibidores , ATPasas de Translocación de Protón/metabolismo , ARN Ribosómico/genética , ARN Ribosómico/metabolismo , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , ARN Nucleolar Pequeño/genética , ARN Nucleolar Pequeño/metabolismo , ARN de Transferencia/genética , ARN de Transferencia/metabolismo , Transducción de SeñalRESUMEN
Alkaline phosphatases (Alps) belong to a class of phosphate transferases that dephosphorylate lipopolysaccharide (LPS), adenosine triphosphate, and nucleotides. In this study, a 1874-base pair (bp) intestinal alp cDNA sequence was cloned from Lateolabrax maculatus and designated as Lm-alpi. It contained a 1611 bp open reading frame which encoded a protein with 537 amino acids. Protein sequence alignment showed that Lm-AlpI shared 29.8-79.8% identity with its homologs. Lm-AlpI catalytic sites contained three metal ion sites (two Zn2+ and one Mg2+), referring to D73, H184, D348, H349, H352, H464, D389, and H390 residues, which are essential for enzymatic activity and conservation in different organisms. Two predicted disulfide bonds in Lm-AlpI were composed of four cysteines (C152-C214 and C499-C506), which were homologous to those of mammals. Immunohistochemical staining revealed that Lm-AlpI was mainly expressed on the mucosal surface of the gastrointestinal tract, including stomach, intestine, and gastric cecum. Lm-AlpI was mainly located on the plasma membrane of transiently transfected HeLa cells. The mRNA of Lm-alpi was mainly expressed in the intestine, and its expression levels gradually increased after LPS treatment and further increased by 1.81-fold after 48 h. After desalting culture, the relative mRNA expression level of Lm-alpi decreased at 30 and 50 days after hatching (DAH) and then returned to normal levels at 70 DAH. Further experiments demonstrated that the enzyme activity of Lm-AlpI exhibited an expression pattern similar to that of the mRNA expression of Lm-alpi after LPS treatment and desalting culture. This study provided valuable information on the Lm-AlpI functions associated with the mucosal immunity and salinity adaptation of L. maculatus.
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Fosfatasa Alcalina/metabolismo , Peces/fisiología , Intestinos/enzimología , Adaptación Fisiológica , Fosfatasa Alcalina/química , Fosfatasa Alcalina/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Enfermedades de los Peces/metabolismo , Enfermedades de los Peces/microbiología , Peces/genética , Regulación Enzimológica de la Expresión Génica/inmunología , Regulación Enzimológica de la Expresión Génica/fisiología , Larva , Filogenia , SalinidadRESUMEN
RATIONALE: Breast cancer is the leading cause of cancer death among women worldwide. Identification of lipid targets that play a role in breast cancer invasion may advance our understanding of the rapid progression of cancer and may lead to the development of new biomarkers for the disease. METHODS: Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI MSI) was applied for the lipidomic profiling of two poorly invasive and two highly invasive breast cancer cell lines to identify the differentially accumulated lipids related to the invasive phenotype. The four cell lines were individually grown on indium tin oxide (ITO)-coated glass slides, analyzed as cell cultures. The raster width and matrix for detection were optimized to improve detection sensitivity. RESULTS: Optimized MSI measurements were performed directly on the cell culture with 9-aminoacridine as matrix, resulting in 215 endogenous compounds detected in positive ion mode and 267 endogenous compounds in negative ion mode in all the four cell lines, representing the largest group of analytes that have been analyzed from cells by a single MSI study. In highly invasive cell lines, 31 lipids including phosphatidylglycerol (PG) and phosphatidic acids were found upregulated and eight lipids including sphingomyelin (SM) downregulated in negative ion mode. The products of de novo fatty acid synthesis incorporated into membrane phospholipids, like oleic-acid-containing PG, may be involved in mitochondrial dysfunction and thus affect the invasion of breast cancer cells. The deficiency of SM may be related to the disruption of apoptosis in highly invasive cancer cells. CONCLUSIONS: This work uncovered more analytes in cells by MSI than previous reports, providing a better visualization and novel insights to advance our understanding of the relationship between rapid progression of breast cancer and lipid metabolism. The most altered lipids may aid the discovery of diagnostic markers and therapeutic targets of breast cancer.
Asunto(s)
Neoplasias de la Mama/química , Lípidos/química , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Biomarcadores/química , Biomarcadores/metabolismo , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular , Femenino , Humanos , Metabolismo de los Lípidos , Invasividad NeoplásicaRESUMEN
We studied molecular events and potential mechanisms underlying the process of female-to-male sex transformation in the rice field eel (Monopterus albus), a protogynous hermaphrodite fish in which the gonad is initially a female ovary and transforms into male testes. We cloned and identified a novel gonadal soma derived factor (GSDF), which encodes a member of the transforming growth factor-beta superfamily. gsdf expression was measured in gonads of female, intersex and male with reverse transcription-PCR and gsdf's role in sex transformation was studied with qPCR, histological analysis and dual-color in situ hybridization assays and compared to other sex-related genes. gsdf was correlated to Sertoli cell differentiation, indicating involvement in testicular differentiation and sex transformation from female to male in this species. A unique expression pattern reveals a potential role of gsdf essential for the sex transformation of rice field eels.
Asunto(s)
Proteínas de Peces/fisiología , Péptidos y Proteínas de Señalización Intercelular/fisiología , Smegmamorpha/fisiología , Animales , Femenino , Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Masculino , Ovario/anatomía & histología , Ovario/crecimiento & desarrollo , Ovario/metabolismo , Procesos de Determinación del Sexo , Diferenciación Sexual , Testículo/anatomía & histología , Testículo/crecimiento & desarrollo , Testículo/metabolismoRESUMEN
Human Aurora kinases, including Aurora kinase A (AURKA), B (AURKB), and C (AURKC), play an essential role in mitotic events such as monitoring of the mitotic checkpoint, creation of bipolar mitotic spindle and alignment of centrosomes on it, also regulating centrosome separation, bio-orientation of chromosomes and cytokinesis. AURKA and AURKB are key regulators of mitosis and centrosome via polymerizing microfilaments and controlling chromatid segregation. In particular, AURKA plays critical roles in the regulation of mitotic entry, centrosome function, bipolar spindle assembly, and chromosome segregation. AURKA has been found to be overexpressed in various solid and haematological cancers and has been linked with poor prognosis. Its important role in cancer initiation, growth, and metastasis has brought the focus to search for potent and selective AURKA inhibitors for cancer treatment. MLN8237, also known as alisertib, is one selective AURKA inhibitor that has shown remarkable anticancer effects in preclinical studies. Alisertib exhibits favourable pharmacokinetic properties. Alisertib has generally showed good partial response rates of 4-52% and good safety profiles in Phase I and II trials when it is solely administered as well as combined with cytotoxic chemotherapeutic drugs. Recently, the multicentre, randomized Phase III study of alisertib in patients with relapsed or refractory peripheral T-cell lymphoma has been discontinued due to unsatisfactory efficacy. The low risk of side effects, accessibility, and effectiveness of alisertib makes it a new promising anticancer therapy and further mechanistic and clinical studies are warranted.
Asunto(s)
Antineoplásicos/farmacocinética , Aurora Quinasa A/antagonistas & inhibidores , Aurora Quinasa A/metabolismo , Inhibidores de Proteínas Quinasas/farmacocinética , Animales , Antineoplásicos/uso terapéutico , Aurora Quinasa A/química , Sitios de Unión/fisiología , Ensayos Clínicos como Asunto/métodos , Humanos , Neoplasias/tratamiento farmacológico , Neoplasias/enzimología , Inhibidores de Proteínas Quinasas/uso terapéutico , Estructura Secundaria de ProteínaRESUMEN
BACKGROUND: Complete mesocolic excision provides a correct anatomical plane for colon cancer surgery. However, manifestation of the surgical plane during laparoscopic complete mesocolic excision versus in computed tomography images remains to be examined. METHODS: Patients who underwent laparoscopic complete mesocolic excision for right-sided colon cancer underwent an abdominal computed tomography scan. The spatial relationship of the intraoperative surgical planes were examined, and then computed tomography reconstruction methods were applied. The resulting images were analyzed. RESULTS: In 44 right-sided colon cancer patients, the surgical plane for laparoscopic complete mesocolic excision was found to be composed of three surgical planes that were identified by computed tomography imaging with cross-sectional multiplanar reconstruction, maximum intensity projection, and volume reconstruction. For the operations performed, the mean bleeding volume was 73±32.3 ml and the mean number of harvested lymph nodes was 22±9.7. The follow-up period ranged from 6-40 months (mean 21.2), and only two patients had distant metastases. CONCLUSIONS: The laparoscopic complete mesocolic excision surgical plane for right-sided colon cancer is composed of three surgical planes. When these surgical planes were identified, laparoscopic complete mesocolic excision was a safe and effective procedure for the resection of colon cancer.