RESUMEN
Trichinella spiralis is an important foodborne zoonotic parasite and it is necessary to develop vaccine to prevent T. spiralis infection in food animals. T. spiralis aspartic protease-2 (TsASP2) has been demonstrated to play a crucial role in larval invasion of intestinal epithelium cells (IECs). The purpose of this study was to assess the interaction between TsASP2 and IECs and to investigate the immune protection elicited by vaccination with rTsASP2. The results showed that the enzymatic activity of native aspartic protease was detected in crude proteins of all T. spiralis development stages other than NBL stage, the highest activity was observed in the IIL stage. The results of Western blot showed that TsASP2 protein was expressed at ML, IIL and AW but not NBL, and the TsASP2 expression level at IIL stage was significantly higher than those of other three worm stages (P < 0.05). The specific binding between rTsASP2 and IECs was observed by immunofluorescence test (IFT) and confocal microscopy, and the binding site was localized at the IEC membrane and this binding ability was inhibited by aspartic protease specific inhibitor pepstain A. The results of ELISA showed that the binding ability was protein dose-dependent. Vaccination with rTsASP2 triggered a mixed Th1/Th2 humoral and mucosal immune responses, as demonstrated by the elevation levels of Th1/Th2 cytokines (IFN-γ and IL-4) secreted by the spleen and mesenteric lymph nodes (MLNs) of immunized mice. The mice vaccinated with rTsASP2 exhibited a 54.17% reduction in enteral adult worms and a 54.58% reduction in muscle larvae after T. spiralis challenge. The results demonstrated that TsASP2 might be a potential molecular target for anti-Trichinella vaccines.
Asunto(s)
Proteasas de Ácido Aspártico/metabolismo , Enterocitos/parasitología , Proteínas del Helminto/metabolismo , Mucosa Intestinal/parasitología , Triquinelosis/parasitología , Animales , Femenino , Inmunidad Humoral , Inmunidad Mucosa , Ratones , Ratones Endogámicos BALB C , Trichinella spiralis/enzimología , Triquinelosis/inmunología , Vacunación , Vacunas/inmunologíaRESUMEN
Based on the density functional theory, the electronic and optical properties of pristine monolayer PdSe2 with Pd or Se vacancy-defect are investigated. Our results show that the Se defect is energetically more favorable than that of Pd defect. The band gap reduces, and some new midgap states appear after the Pd or Se defects are introduced. In terms of the optical properties, the prominent anisotropic characters are remained. The obvious new peaks of the dielectric constant appear after introducing defects. The light absorption in the visible energy range expands based on the appearance of the midgap states induced by the Pd or Se defects. The changes of the refractive index and reflectivity are similar with those of the dielectric constants and the light absorption. The energy loss spectrum of the PdSe2 with Pd or Se defects is obviously different, which can be used to identify different defects in PdSe2. These findings provide effective strategies to tune electronic and optical properties of monolayer PdSe2 by introducing defects.
RESUMEN
The zinc-finger protein tristetraprolin (TTP) binds to AU-rich elements present in the 3' untranslated regions of transcripts that mainly encode proteins of the inflammatory response. TTP-bound mRNAs are targeted for destruction via recruitment of the eight-subunit deadenylase complex "carbon catabolite repressor protein 4 (CCR4)-negative on TATA-less (NOT)," which catalyzes the removal of mRNA poly-(A) tails, the first obligatory step in mRNA decay. Here we show that a novel interaction between TTP and the CCR4-NOT subunit, CNOT9, is required for recruitment of the deadenylase complex. In addition to CNOT1, CNOT9 is now included in the identified CCR4-NOT subunits shown to interact with TTP. We find that both the N- and C-terminal domains of TTP are involved in an interaction with CNOT9. Through a combination of SPOT peptide array, site-directed mutagenesis, and bio-layer interferometry, we identified several conserved tryptophan residues in TTP that serve as major sites of interaction with two tryptophan-binding pockets of CNOT9, previously found to interact with another modulator GW182. We further demonstrate that these interactions are also required for recruitment of the CCR4-NOT complex and TTP-directed decay of an mRNA containing an AU-rich element in its 3'-untranslated region. Together the results reveal new molecular details for the TTP-CNOT interaction that shape an emerging mechanism whereby TTP targets inflammatory mRNAs for deadenylation and decay.
Asunto(s)
Factores de Transcripción/metabolismo , Tristetraprolina/metabolismo , Triptófano/metabolismo , Regiones no Traducidas 3' , Autoantígenos/genética , Autoantígenos/metabolismo , Exorribonucleasas/genética , Exorribonucleasas/metabolismo , Células HeLa , Humanos , Mutagénesis Sitio-Dirigida , Dominios y Motivos de Interacción de Proteínas , Estabilidad del ARN , ARN Mensajero/metabolismo , Proteínas de Unión al ARN/genética , Proteínas de Unión al ARN/metabolismo , Receptores CCR4/genética , Receptores CCR4/metabolismo , Factores de Transcripción/genética , Tristetraprolina/genética , Triptófano/genéticaRESUMEN
Perrault syndrome is a rare autosomal recessive disorder characterized by sensorineural hearing loss (SNHL) in both sexes and primary ovarian insufficiency in 46, XX karyotype females. Biallelic variants in five genes are reported to be causative: HSD17B4, HARS2, LARS2, CLPP and C10orf2. Here we present eight families affected by Perrault syndrome. In five families we identified novel or previously reported variants in HSD17B4, LARS2, CLPP and C10orf2. The proband from each family was whole exome sequenced and variants confirmed by Sanger sequencing. A female was compound heterozygous for a known, p.(Gly16Ser) and novel, p.(Val82Phe) variant in D-bifunctional protein (HSD17B4). A family was homozygous for mitochondrial leucyl aminocyl tRNA synthetase (mtLeuRS) (LARS2) p.(Thr522Asn), previously associated with Perrault syndrome. A further family was compound heterozygous for mtLeuRS, p.(Thr522Asn) and a novel variant, p.(Met117Ile). Affected individuals with LARS2 variants had low frequency SNHL, a feature previously described in Perrault syndrome. A female with significant neurological disability was compound heterozygous for p.(Arg323Gln) and p.(Asn399Ser) variants in Twinkle (C10orf2). A male was homozygous for a novel variant in CLPP, p.(Cys144Arg). In three families there were no putative pathogenic variants in these genes confirming additional disease-causing genes remain unidentified. We have expanded the spectrum of disease-causing variants associated with Perrault syndrome.
Asunto(s)
Aminoacil-ARNt Sintetasas/genética , ADN Helicasas/genética , Endopeptidasa Clp/genética , Disgenesia Gonadal 46 XX/genética , Pérdida Auditiva Sensorineural/genética , Proteínas Mitocondriales/genética , Proteína-2 Multifuncional Peroxisomal/genética , Exoma/genética , Femenino , Genotipo , Disgenesia Gonadal 46 XX/patología , Pérdida Auditiva Sensorineural/patología , Homocigoto , Humanos , Masculino , Mutación , Linaje , Fenotipo , Insuficiencia Ovárica Primaria/genética , Insuficiencia Ovárica Primaria/fisiopatologíaRESUMEN
OBJECTIVE: To observe the heart rate turbulence (HRT) in patients with masked hypertension (MH), and white-coat hypertension (WCH). PATIENTS AND METHODS: Patients were classified on the basis of clinic and 24h ambulatory blood-pressure monitoring: essential hypertension (H, n = 32), masked hypertension (MH, n=26), white-coat hypertension (WCH, n = 29) and normotension (NT, n = 30). For each subject, we recorded 24 hours holter monitoring electrocardiogram, calculated the turbulence onset (TO) and turbulence slope (TS) and compared the differences. RESULTS: Compared with NT controls, the differences of TO and TS in the patients with EH, MH and WCH were statistically significant (p < 0.01). No significant differences were found between the EH, MH and WCH groups. CONCLUSIONS: The HRT in EH, MH and WCH patients is significantly lower, when their autonomic nerve function is damaged.
Asunto(s)
Monitoreo Ambulatorio de la Presión Arterial/métodos , Frecuencia Cardíaca , Hipertensión Enmascarada/diagnóstico , Hipertensión Enmascarada/fisiopatología , Hipertensión de la Bata Blanca/diagnóstico , Hipertensión de la Bata Blanca/fisiopatología , Anciano , Femenino , Frecuencia Cardíaca/fisiología , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Post-translational modification of chromatin is emerging as an increasingly important regulator of chromosomal processes. In particular, histone lysine and arginine methylation play important roles in regulating transcription, maintaining genomic integrity, and contributing to epigenetic memory. Recently, the use of new approaches to analyse histone methylation, the generation of genetic model systems, and the ability to interrogate genome wide histone modification profiles has aided in defining how histone methylation contributes to these processes. Here we focus on the recent advances in our understanding of the histone methylation system and examine how dynamic histone methylation contributes to normal cellular function in mammals.