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1.
Front Cardiovasc Med ; 9: 1052094, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36523362

RESUMEN

Objective: Despite the rapid development of thoracic endovascular aortic repair (TEVAR), it is still a challenge to maintain the blood flow of the branch arteries above the aortic arch in Stanford type B aortic dissection involving the left subclavian artery (LSA). The Castor stent graft is an integrated, customized, single-branch stent that enables reconstruction of the LSA. The purpose of this systematic review and meta-analysis was to assess the efficacy of the Castor stent graft for type B aortic dissection. Materials and methods: An extensive electronic literature search (PROSPERO registration number: CRD42022322146) was undertaken to identify all articles published up to August 2022 that described thoracic aortic repair with branch stents in the treatment of type B aortic dissection involving the LSA. The quality of the included studies was analyzed using the MINORS criteria. The primary outcome measures were the technical success rate, early mortality rate, endoleak rate, and 1-year survival rate. The secondary outcome measures were the stroke rate, left upper extremity ischemia rate, and target vessel patency rate. Results: Eleven studies involving 415 patients were eligible for this meta-analysis. The LSA was successfully preserved in all procedures. The technical success rate was 97.5% (95% CI: 0.953-0.991); the intraoperative endoleak rate was 0.1% (95% CI: 0.000-0.012); the intraoperative LSA patency rate was 99.52%; the intraoperative LSA stent deformation and stenosis rate was 0.15% (95% CI: 0.000-0.051); the early type I endoleak rate was 1.6% (95% CI: 0.003-0.035); the 30-day mortality rate was 0.96%; the early reintervention rate was 0.9% (95% CI: 0.000-0.040); and the perioperative stroke rate was 0% (95% CI: 0.000-0.005). The 1-year survival rate was 99.7% (95% CI: 0.976-1.000). The half-year LSA patency rate was 99.3%, the 1-year LSA patency rate was 97.58%, and the 2-year LSA patency rate was 95.23%. During the follow-up period, the leakage rate was 0.3% (95% CI: 0.000-0.017), the incidence of left upper extremity ischemia rate was 0.5% (95% CI: 0.000-0.035), and the deformation and stenosis rate of the LSA stent was 2.2% (95% CI: 0.06-0.046). Conclusion: This meta-analysis shows that endovascular repair of type B aortic dissection using the Castor stent-graft may be technically feasible and effective. However, this conclusion needs to be interpreted with caution, as the quality of evidence for all outcomes is between low and very low. Systematic review registration: [https://www.crd.york.ac.uk/prospero/], identifier [CRD42022322146].

2.
Bull Entomol Res ; 112(2): 187-196, 2022 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-34474703

RESUMEN

Diadegma semiclausum is an important parasitoid wasp and widely used in the biological control of the diamondback moth, Plutella xylostella, one of the most destructive pests of cruciferous plants. Insect olfactory system is critical in guiding behaviors including feeding, mating, and oviposition, in which odorant binding proteins (OBPs) and odorant receptors (ORs) are two key components. However, limited attention has been paid to D. semiclausum olfactory genes. In this study, a transcriptome sequencing was performed on the RNA samples extracted from D. semiclausum male and female adult antennae. A total of 17 putative OBP and 67 OR genes were annotated and further compared to OBPs and ORs from P. xylostella, and other hemipteran parasitoid species. The expression patterns of D. semiclausum OBPs between male and female antennae were examined using reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR. Six OBPs (DsemOBP 6, 7, 8, 9, 10, and 14) demonstrated significantly higher expression levels in females than in males, which may assist in female D. semiclausum host-seeking and oviposition behaviors. This study advances our understanding of the olfactory system of D. semiclausum at the molecular level and paves the way for future functional studies aiming at increasing the efficacy to control P. xylostella by using D. semiclausum.


Asunto(s)
Mariposas Nocturnas , Avispas , Animales , Antenas de Artrópodos/metabolismo , Femenino , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Larva/fisiología , Masculino , Mariposas Nocturnas/genética , Oviposición , Olfato , Avispas/genética
3.
Insect Sci ; 28(3): 769-779, 2021 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32420694

RESUMEN

Sensory neuron membrane proteins (SNMPs) play a critical role in insect chemosensory system. Previously, three SNMPs were identified, characterized and functionally investigated in a lepidopteran model insect, Bombyx mori. However, whether these results are consistent across other lepidopteran species are unknown. Here genome and transcriptome data analysis, expression profiling, quantitative real-time PCR (qRT-PCR) and the yeast hybridization system were utilized to examine snmp genes of Helicoverpa armigera, one of the most destructive lepidopteran pests in cropping areas. In silico expression and qRT-PCR analyses showed that, just as the B. mori snmp genes, H. armigera snmp1 (Harmsnmp1) is specifically expressed in adult antennae. Harmsnmp2 is broadly expressed in multiple tissues including adult antennae, tarsi, larval antennae and mouthparts. Harmsnmp3 is specifically expressed in larval midguts. Further RNAseq analysis suggested that the expression levels of Harmsnmp2 and Harmsnmp3 differed significantly depending on the plant species on which the larvae fed, indicating they may be involved in plant-feeding behaviours. Yeast hybridization results revealed a protein-protein interaction between HarmSNMP1 and the sex pheromone receptor, HarmOR13. This study demonstrated that SNMPs may share same functions and mechanisms in different lepidopteran species, which improved our understanding of insect snmp genes and their functions in lepidopterans.


Asunto(s)
Mariposas Nocturnas , Neuronas Receptoras Olfatorias/metabolismo , Receptores de Feromonas , Animales , Antenas de Artrópodos/metabolismo , Conducta Alimentaria , Perfilación de la Expresión Génica , Genes de Insecto , Genoma de los Insectos , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Mariposas Nocturnas/genética , Mariposas Nocturnas/metabolismo , Filogenia , Receptores de Feromonas/genética , Receptores de Feromonas/metabolismo , Células Receptoras Sensoriales/metabolismo , Atractivos Sexuales/metabolismo , Transcriptoma
4.
Comput Biol Med ; 119: 103687, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-32339122

RESUMEN

Recent advances in data analytics and computer-aided diagnostics stimulate the vision of patient-centric precision healthcare, where treatment plans are customized based on the health records and needs of every patient. In physical rehabilitation, the progress in machine learning and the advent of affordable and reliable motion capture sensors have been conducive to the development of approaches for automated assessment of patient performance and progress toward functional recovery. The presented study reviews computational approaches for evaluating patient performance in rehabilitation programs using motion capture systems. Such approaches will play an important role in supplementing traditional rehabilitation assessment performed by trained clinicians, and in assisting patients participating in home-based rehabilitation. The reviewed computational methods for exercise evaluation are grouped into three main categories: discrete movement score, rule-based, and template-based approaches. The review places an emphasis on the application of machine learning methods for movement evaluation in rehabilitation. Related work in the literature on data representation, feature engineering, movement segmentation, and scoring functions is presented. The study also reviews existing sensors for capturing rehabilitation movements and provides an informative listing of pertinent benchmark datasets. The significance of this paper is in being the first to provide a comprehensive review of computational methods for evaluation of patient performance in rehabilitation programs.


Asunto(s)
Terapia por Ejercicio , Movimiento , Ejercicio Físico , Humanos
5.
IEEE Trans Neural Syst Rehabil Eng ; 28(2): 468-477, 2020 02.
Artículo en Inglés | MEDLINE | ID: mdl-31940544

RESUMEN

Computer-aided assessment of physical rehabilitation entails evaluation of patient performance in completing prescribed rehabilitation exercises, based on processing movement data captured with a sensory system. Despite the essential role of rehabilitation assessment toward improved patient outcomes and reduced healthcare costs, existing approaches lack versatility, robustness, and practical relevance. In this paper, we propose a deep learning-based framework for automated assessment of the quality of physical rehabilitation exercises. The main components of the framework are metrics for quantifying movement performance, scoring functions for mapping the performance metrics into numerical scores of movement quality, and deep neural network models for generating quality scores of input movements via supervised learning. The proposed performance metric is defined based on the log-likelihood of a Gaussian mixture model, and encodes low-dimensional data representation obtained with a deep autoencoder network. The proposed deep spatio-temporal neural network arranges data into temporal pyramids, and exploits the spatial characteristics of human movements by using sub-networks to process joint displacements of individual body parts. The presented framework is validated using a dataset of ten rehabilitation exercises. The significance of this work is that it is the first that implements deep neural networks for assessment of rehabilitation performance.


Asunto(s)
Aprendizaje Profundo , Terapia por Ejercicio/métodos , Rehabilitación/métodos , Algoritmos , Automatización , Fenómenos Biomecánicos , Simulación por Computador , Voluntarios Sanos , Humanos , Movimiento/fisiología , Redes Neurales de la Computación , Distribución Normal , Resultado del Tratamiento
6.
Mol Nutr Food Res ; 62(11): e1701050, 2018 06.
Artículo en Inglés | MEDLINE | ID: mdl-29644801

RESUMEN

SCOPE: This study investigates the ability of preterm milk exosomes to survive gastric/pancreatic digestion, internalization by intestinal epithelia, and the microRNAs (miRNAs) contents. METHODS AND RESULTS: At average infant age 1 week and 6 days, milk is collected from mothers who delivered preterm and term infants (n = 10). Milk is exposed to conditions simulating infant gut digestion. Exosomes are isolated and lysed, and the exposed miRNAs are sequenced. Preterm milk exosomes survive in vitro digestion, and can be taken up by intestinal epithelia. Three hundred and thirty miRNAs are identified as preterm milk exosome miRNAs, and in vitro digestion does not have a pronounced effect on their expression. The abundant miRNAs in preterm milk exosomes are similar to those from term milk. Twenty-one low abundance miRNAs are specifically expressed in preterm milk exosomes compared to early term milk in the current study and what previously is found in mature term milk. CONCLUSION: These results for the first time reveal the survivability of preterm milk exosomes following simulated gastric/pancreatic digestion. The authors demonstrate the richness of the miRNAs content in these exosomes. The results improve the knowledge of preterm milk biology and the molecular basis by which exosome miRNAs may uniquely affect preterm infants during early development.


Asunto(s)
Exosomas/genética , Intestino Delgado/metabolismo , MicroARNs/análisis , Leche Humana/citología , Adulto , Digestión , Células Epiteliales/metabolismo , Exosomas/metabolismo , Femenino , Humanos , Recién Nacido , Recien Nacido Prematuro , Intestino Delgado/citología , Madres
7.
Naturwissenschaften ; 104(11-12): 94, 2017 Oct 23.
Artículo en Inglés | MEDLINE | ID: mdl-29063281

RESUMEN

Aldehyde oxidases (AOXs) are a family of metabolic enzymes that oxidize aldehydes into carboxylic acids; therefore, they play critical roles in detoxification and degradation of chemicals. By using transcriptomic and genomic approaches, we successfully identified six putative AOX genes (HarmAOX1-6) from cotton bollworm, Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). In silico expression profile, reverse transcription (RT)-PCR, and quantitative PCR (qPCR) analyses showed that HarmAOX1 is highly expressed in adult antennae, tarsi, and larval mouthparts, so they may play an important role in degrading plant-derived compounds. HarmAOX2 is highly and specifically expressed in adult antennae, suggesting a candidate pheromone-degrading enzyme (PDE) to inactivate the sex pheromone components (Z)-11-hexadecenal and (Z)-9-hexadecenal. RNA sequencing data further demonstrated that a number of host plants they feed on could significantly upregulate the expression levels of HarmAOX1 in larvae. This study improves our understanding of insect aldehyde oxidases and insect-plant interactions.


Asunto(s)
Aldehído Oxidasa/genética , Aldehído Oxidasa/metabolismo , Regulación Enzimológica de la Expresión Génica , Mariposas Nocturnas/enzimología , Mariposas Nocturnas/genética , Animales , Perfilación de la Expresión Génica , Larva , Plantas/química
8.
J Proteome Res ; 16(11): 4113-4121, 2017 11 03.
Artículo en Inglés | MEDLINE | ID: mdl-28925267

RESUMEN

Whey proteins and caseins in breast milk provide bioactivities and also have different amino acid composition. Accurate determination of these two major protein classes provides a better understanding of human milk composition and function, and further aids in developing improved infant formulas based on bovine whey proteins and caseins. In this study, we implemented a LC-MS/MS quantitative analysis based on iBAQ label-free quantitation, to estimate absolute concentrations of α-casein, ß-casein, and κ-casein in human milk samples (n = 88) collected between day 1 and day 360 postpartum. Total protein concentration ranged from 2.03 to 17.52 with a mean of 9.37 ± 3.65 g/L. Casein subunits ranged from 0.04 to 1.68 g/L (α-), 0.04 to 4.42 g/L (ß-), and 0.10 to 1.72 g/L (α-), with ß-casein having the highest average concentration among the three subunits. Calculated whey/casein ratio ranged from 45:55 to 97:3. Linear regression analyses show significant decreases in total protein, ß-casein, κ-casein, total casein, and a significant increase of whey/casein ratio during the course of lactation. Our study presents a novel and accurate quantitative analysis of human milk casein content, demonstrating a lower casein content than earlier believed, which has implications for improved infants formulas.


Asunto(s)
Caseínas/análisis , Leche Humana/química , Proteína de Suero de Leche/análisis , Humanos , Lactancia , Proteínas de la Leche/análisis , Suero Lácteo
9.
Genome ; 60(12): 1037-1044, 2017 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-28825966

RESUMEN

Insects utilize sugars as their essential energy and nutrient sources; therefore, the sense of sugar detection plays a critical role in insect behaviours. Previously, using genomic and transcriptomic approaches, we identified eight putative sugar gustatory receptor (GR) genes from the cotton bollworm Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae). Here, we further validated these annotated sugar receptor genes (HarmGr4-HarmGr8 and HarmGr10-HarmGr12) and found HarmGr10 may be a pseudogene carrying a stop codon in the open reading frame. Sequence alignment revealed H. armigera sugar GR sequences are conserved at C-terminus and phylogenetic analysis showed that insect sugar GRs have evolved in a family-specific manner. Interestingly, all eight H. armigera sugar GRs are localized in a tandem array on the same scaffold of the genome. In silico gene expression and reverse transcription (RT)-PCR analysis showed that HarmGr10 is specifically expressed in male adult testes while HarmGr11 is specifically expressed in female adult ovaries, suggesting H. armigera sugar GRs may be involved in reproduction-related functions. This study improves our knowledge on insect sugar receptors and gustatory systems.


Asunto(s)
Proteínas de Insectos/genética , Lepidópteros/genética , Receptores de Superficie Celular/genética , Azúcares/metabolismo , Gusto , Animales , Secuencia Conservada , Evolución Molecular , Femenino , Proteínas de Insectos/química , Proteínas de Insectos/metabolismo , Lepidópteros/clasificación , Masculino , Ovario/metabolismo , Filogenia , Dominios Proteicos , Seudogenes , Receptores de Superficie Celular/química , Receptores de Superficie Celular/metabolismo , Testículo/metabolismo
10.
Mol Nutr Food Res ; 61(11)2017 11.
Artículo en Inglés | MEDLINE | ID: mdl-28688106

RESUMEN

SCOPE: Human milk exosomes provide a natural means of genetic material transfer to infants; however, the effect of gastric/pancreatic digestion milk exosomes stability and their microRNA content is largely unknown. METHODS AND RESULTS: We took a simulated gastric/pancreatic digestion protocol to perform in vitro digestion of milk exosomes, explore intestinal epithelial uptake, and further elucidate microRNA responses to digestion at early-, mid-, late lactation by massive parallel sequencing. Both undigested and digested exosomes enter human intestinal crypt-like cells (HIEC), with evidence of nuclear localization. We identified 288 mature microRNAs from all 24 exosome samples, and an additional 610 at low abundance. A large number of synapse development- and immune-related microRNAs were identified. hsa-miR-22-3p was the most abundant microRNA, and the top 15 microRNAs contributed ∼11% of the sequencing reads. Upon digestion, the overall microRNA abundance in human milk exosomes was stable. CONCLUSION: Our results for the first time reveal the survivability and complexity of human milk exosome microRNAs upon simulated gastric/pancreatic digestion, and the dynamics during lactation stages. The results suggest a previously underexplored area of infant response to genetic material in human milk exosomes.


Asunto(s)
Digestión , Enterocitos/metabolismo , Exosomas/metabolismo , Absorción Intestinal , MicroARNs/metabolismo , Proteínas de la Leche/metabolismo , Leche Humana/metabolismo , Transporte Activo de Núcleo Celular , Adulto , Secuencia de Bases , Línea Celular , Células Cultivadas , Enterocitos/citología , Exosomas/química , Exosomas/ultraestructura , Femenino , Humanos , Lactancia , MicroARNs/química , Microscopía Electrónica de Rastreo , Proteínas de la Leche/química , Leche Humana/química , Tamaño de la Partícula , Estabilidad Proteica , Estabilidad del ARN , Análisis de Secuencia de ARN , Adulto Joven
11.
J Nutr Biochem ; 37: 101-108, 2016 11.
Artículo en Inglés | MEDLINE | ID: mdl-27697743

RESUMEN

Human milk contains growth factors that maintain intestinal mucosal homeostasis, but the molecular mechanisms behind how these growth factors regulate gene transcription are largely unknown. In this study, IEC-6 (rat intestinal epithelial cells) cells were used as a model to study cell differentiation mediated by transforming growth factor-ß2 (TGF-ß2), the most abundant growth factor in human milk. We focused on the transcription factor early growth response-1 (EGR-1), as we found a robust and rapid response in our initial transcription factor screen. Immunoblotting and immunofluorescent assays confirmed the phenotype change upon TGF-ß2 treatment and EGR-1 stimulation in the nucleus, with maximum expression occurring at 1 h. Chromatin immunoprecipitation sequencing was performed to map genome-wide EGR-1 binding sites on more than 1800 genes, widely involved in processes such as gene expression, transcription, membrane invagination and metabolism. In particular, more than 15 Wnt signaling pathway genes have EGR-1 binding sites; among them, Axin1 was the limiting factor, ensuring proper ß-catenin accumulation in the cytoplasm. We further used chromatin immunoprecipitation quantitative PCR to validate that EGR-1 binds to the region of -636/-454 bp and -454/-200 bp of the Axin1 promoter and functionally activates gene expression. The effect of TGF-ß2 on maintaining small intestinal cell homeostasis was partially explained by Axin1 activation through EGR-1.


Asunto(s)
Proteína Axina/agonistas , Diferenciación Celular , Proteína 1 de la Respuesta de Crecimiento Precoz/metabolismo , Mucosa Intestinal/metabolismo , Intestino Delgado/metabolismo , Receptores de Factores de Crecimiento Transformadores beta/agonistas , Factor de Crecimiento Transformador beta2/metabolismo , Transporte Activo de Núcleo Celular , Animales , Proteína Axina/genética , Proteína Axina/metabolismo , Línea Celular , Inmunoprecipitación de Cromatina , Biología Computacional , Proteína 1 de la Respuesta de Crecimiento Precoz/antagonistas & inhibidores , Proteína 1 de la Respuesta de Crecimiento Precoz/genética , Regulación del Desarrollo de la Expresión Génica , Genes Reporteros , Células HEK293 , Humanos , Mucosa Intestinal/citología , Intestino Delgado/citología , Regiones Promotoras Genéticas , Proteínas Serina-Treonina Quinasas/metabolismo , Interferencia de ARN , Ratas , Receptor Tipo II de Factor de Crecimiento Transformador beta , Receptores de Factores de Crecimiento Transformadores beta/metabolismo , Elementos de Respuesta , Transducción de Señal , Proteínas Wnt/agonistas , Proteínas Wnt/genética , Proteínas Wnt/metabolismo
12.
Plant Cell ; 28(10): 2435-2452, 2016 10.
Artículo en Inglés | MEDLINE | ID: mdl-27729395

RESUMEN

RNA binding proteins (RBPs) control the fate and expression of a transcriptome. Despite this fundamental importance, our understanding of plant RBPs is rudimentary, being mainly derived via bioinformatic extrapolation from other kingdoms. Here, we adapted the mRNA-protein interactome capture method to investigate the RNA binding proteome in planta. From Arabidopsis thaliana etiolated seedlings, we captured more than 700 proteins, including 300 with high confidence that we have defined as the At-RBP set. Approximately 75% of these At-RBPs are bioinformatically linked with RNA biology, containing a diversity of canonical RNA binding domains (RBDs). As no prior experimental RNA binding evidence exists for the majority of these proteins, their capture now authenticates them as RBPs. Moreover, we identified protein families harboring emerging and potentially novel RBDs, including WHIRLY, LIM, ALBA, DUF1296, and YTH domain-containing proteins, the latter being homologous to animal RNA methylation readers. Other At-RBP set proteins include major signaling proteins, cytoskeleton-associated proteins, membrane transporters, and enzymes, suggesting the scope and function of RNA-protein interactions within a plant cell is much broader than previously appreciated. Therefore, our foundation data set has provided an unbiased insight into the RNA binding proteome of plants, on which future investigations into plant RBPs can be based.


Asunto(s)
Arabidopsis/metabolismo , Proteoma/metabolismo , ARN Mensajero/metabolismo , Proteínas de Unión al ARN/metabolismo , Plantones/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Unión al ARN/genética , Plantones/genética
13.
Pediatr Res ; 80(5): 759-765, 2016 11.
Artículo en Inglés | MEDLINE | ID: mdl-27603563

RESUMEN

BACKGROUND: Growth factors (GFs) are milk bioactive components contributing to the regulation of neonatal small intestinal maturation, and their receptors on the small intestinal epithelium play essential roles in mediating the functions of GFs. There is limited data correlating milk GFs and their receptors in the neonatal small intestine during the perinatal period. METHODS: Small intestines of C57BL/6N mouse pups were collected at regular intervals during fetal life and up to postnatal day (PD) 60. Gene expression of GF receptors was determined by real-time qPCR. Milk GF concentrations up to PD21 were analyzed by enzyme-linked immunosorbent assay. RESULTS: The majority of GF receptors showed significantly greater expression in the fetus than in postnatal life, and a sharp decrease occurred from PD14 extending to PD60; solid food restriction (PD14 and PD18) did not affect this decrease. Concentrations of five detected milk GFs demonstrated that GFs and the corresponding small intestinal receptors exhibited different correlations, with only milk transforming growth factor ß1 (TGF-ß1) having a significant positive correlation with TGF-ß receptor 1 mRNA. CONCLUSION: Gene expression of small intestinal GF receptors is likely a process of neonatal intestinal maturation that is affected concurrently by milk GFs and additional endogenous factors.


Asunto(s)
Péptidos y Proteínas de Señalización Intercelular/metabolismo , Intestino Delgado/metabolismo , Leche/química , Animales , Animales Recién Nacidos , Receptores ErbB/sangre , Femenino , Regulación del Desarrollo de la Expresión Génica , Intestino Delgado/embriología , Intestino Delgado/crecimiento & desarrollo , Lactancia , Masculino , Ratones , Ratones Endogámicos C57BL , Proteínas Proto-Oncogénicas c-met/sangre , Receptor IGF Tipo 1/sangre , Receptor de Insulina/sangre , Receptor de Factor de Crecimiento Nervioso/sangre , Receptores de Factores de Crecimiento de Fibroblastos/sangre , Receptores del Factor de Crecimiento Derivado de Plaquetas/sangre , Receptores de Factores de Crecimiento Transformadores beta/sangre , Receptor 1 de Factores de Crecimiento Endotelial Vascular/sangre
14.
Cell Rep ; 16(5): 1456-1469, 2016 08 02.
Artículo en Inglés | MEDLINE | ID: mdl-27452465

RESUMEN

RNA functions through the dynamic formation of complexes with RNA-binding proteins (RBPs) in all clades of life. We determined the RBP repertoire of beating cardiomyocytic HL-1 cells by jointly employing two in vivo proteomic methods, mRNA interactome capture and RBDmap. Together, these yielded 1,148 RBPs, 391 of which are shared with all other available mammalian RBP repertoires, while 393 are thus far unique to cardiomyocytes. RBDmap further identified 568 regions of RNA contact within 368 RBPs. The cardiomyocyte mRNA interactome composition reflects their unique biology. Proteins with roles in cardiovascular physiology or disease, mitochondrial function, and intermediary metabolism are all highly represented. Notably, we identified 73 metabolic enzymes as RBPs. RNA-enzyme contacts frequently involve Rossmann fold domains with examples in evidence of both, mutual exclusivity of, or compatibility between RNA binding and enzymatic function. Our findings raise the prospect of previously hidden RNA-mediated regulatory interactions among cardiomyocyte gene expression, physiology, and metabolism.


Asunto(s)
Miocitos Cardíacos/metabolismo , Proteoma/metabolismo , ARN Mensajero/metabolismo , Proteínas de Unión al ARN/metabolismo , Línea Celular Tumoral , Células HeLa , Cardiopatías/metabolismo , Humanos , Proteómica/métodos
15.
Vet Microbiol ; 166(3-4): 356-64, 2013 Oct 25.
Artículo en Inglés | MEDLINE | ID: mdl-23876931

RESUMEN

Cases of myeloid leukosis and hemangioma associated with avian leukosis virus subgroup J (ALV-J) are becoming more frequent in China in commercial layer chickens and breeders of egg-type chickens. In this study, two strains of ALV-J (SCAU11-H and SCAU11-XG) associated with hemangioma and myelocytoma were isolated from commercial broiler breeder animals in 2011. Their full-length proviral sequences were analyzed, revealing several unique genetic differences between the two isolates, and suggesting that the two viruses were derived from two distinct lineages. Strain SCAU11-H showed high sequence homology to early Chinese isolates associated with hemangioma, while strain SCAU11-XG was genetically closer to the prototype strain, HPRS-103. The complete genomic nucleotide sequences of SCAU11-H and SCAU11-XG were 7471 bp and 7727 bp in length, respectively. They shared 94.8% identity with each other, and had 94.0-96.8% nucleotide identity to ALV-J reference isolates. Homology analysis of the env, pol, and gag genes of the two isolates and other references strains showed that the gag and pol genes of the two viruses were more conserved than the env gene. In addition, the two isolates had significant deletions and substitutions in their 3'-UTR regions, compared to HPRS-103. These results suggest that the env gene and the 3'-UTR regions in these ALV-J isolates have evolved rapidly, and might be involved in the oncogenic spectrum of ALV-J. The results of this study contribute to our further study of the relationship between ALV integration patterns and multi-pathotypes associated with ALV-J.


Asunto(s)
Virus de la Leucosis Aviar/clasificación , Virus de la Leucosis Aviar/aislamiento & purificación , Leucosis Aviar/virología , Hemangioma/veterinaria , Leucemia Mieloide/veterinaria , Filogenia , Enfermedades de las Aves de Corral/virología , Animales , Virus de la Leucosis Aviar/genética , Secuencia de Bases , Pollos/genética , Pollos/virología , China , Hemangioma/virología , Leucemia Mieloide/virología , Datos de Secuencia Molecular
16.
Genes Nutr ; 8(1): 69-78, 2013 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22570175

RESUMEN

TGF-ß is a potent pleiotropic factor that promotes small intestinal cell differentiation. The role of microRNAs in the TGF-ß induction of intestinal epithelial phenotype is largely unknown. We hypothesized that microRNAs are functionally involved in TGF-ß-induced intestinal cell growth. In this study, TGF-ß caused a morphological change of IEC-6 cells and stimulated expression of the epithelial cell markers alkaline phosphatase, villin, and aminopeptidase N. By global microRNA profiling during TGF-ß-induced intestinal crypt cell (IEC-6) differentiation, we identified 19 differentially expressed microRNAs. We showed by real-time Q-PCR that miR-146b expression increased rapidly after TGF-ß treatment; sequence analysis and in vitro assays revealed that miR-146b targets SIAH2, an E3 ubiquitin ligase, with decreased protein expression upon IEC-6 cell differentiation. Transfection of miR-146b inhibitor before TGF-ß treatment blocked the down-regulation of SIAH2 in response to TGF-ß. Moreover, SIAH2 over-expression during TGF-ß treatment caused a significant decrease in Smad7 protein expression in IEC-6 cells. Furthermore, activation of the ERK1/2 pathway is active in the up-regulation of miR-146b by TGF-ß. These findings suggest a novel mechanism whereby TGF-ß signaling during IEC-6 cell differentiation may be modulated in part by microRNAs, and we propose a key role for miR-146b in the homeostasis of growth factor TGF-ß signaling through a negative feedback regulation involving down-regulation of SIAH2 repressed Smad7 activities.

17.
Biochem Cell Biol ; 90(3): 476-84, 2012 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-22332905

RESUMEN

Postnatal modeling of the intestinal epithelium has long-term impacts on the healthy development of infants and relies largely on nutrient composition of the diet. Lactoferrin (Lf) is among the various human milk trophic factors that facilitate the infant intestinal adaptation. Hydrolysis of Lf is minimal at the prevailing postprandial pH of infants, and Lf may therefore have greater biological potential in infants than in adults. Lf bidirectionally stimulates concentration-dependent proliferation and differentiation of small intestinal epithelial cells, and therefore affects small intestinal mass, length, and epithelial digestive enzyme expression. A 105 kDa Lf receptor (LfR) specifically mediates the uptake of Lf into enterocytes and crypt cells. Mechanistically, the complex of Lf and LfR is internalized through clathrin-mediated endocytosis; both iron-free apo-Lf and iron-saturated holo-Lf activate the PI3K/Akt pathway, whereas only apo-Lf triggers ERK1/2 signaling. Lf enters the nucleus, where it can stimulate thymidine incorporation into crypt cells, regulating transcription of genes such as TGF-ß1. In the fetus, the plasma membrane LfR is at the highest abundance in the small intestine, and the receptor gene is tightly controlled at multiple levels. Aspecific microRNA, miR-584, is involved in the posttranscriptional regulation of LfR, and in the human LfR DNA promoter, 2 Sp1 binding sites have been characterized functionally. Finally, cell proliferation and global gene expression reveal that native bovine Lf can perform biological activities similar to those exerted by human Lf in postnatal small intestinal development.


Asunto(s)
Intestino Delgado/crecimiento & desarrollo , Lactoferrina/fisiología , Animales , Diferenciación Celular , Proliferación Celular , Regulación del Desarrollo de la Expresión Génica , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/fisiología , Intestino Delgado/citología , Intestino Delgado/metabolismo , Lactoferrina/genética , Lactoferrina/metabolismo , Receptores de Superficie Celular/metabolismo
18.
J Med Microbiol ; 61(Pt 3): 375-383, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22052996

RESUMEN

Lactoferrin has been suggested to have antiviral activity against hepatitis C virus (HCV). The objective of this study was to compare the effects of recombinant camel lactoferrin (rcLf), native camel lactoferrin (ncLf) and their N and C fragments on HCV infection in Huh7.5 cells. ncLf was purified from camel milk and N and C lobes were generated proteolytically. rcLf and its fragments were synthesized using a Bac-to-Bac baculovirus expression system. All proteins except the C lobe of rcLf were soluble. The inhibitory effects on HCV entry into Huh7.5 cells were evaluated by incubation of HCV with Lf prior to infection or pre-treatment of the cells with Lf prior to infection. The inhibitory effect on HCV amplification in Huh7.5 cells was determined by Lf treatment of HCV-infected cells. Nested RT-PCR was performed to amplify intracellular HCV 5' non-coding RNA sequences. rcLf and ncLf and their fragments could prevent HCV entry into Huh7.5 cells by direct interaction with the virus and inhibited virus amplification in Huh7.5 cells. Therefore, the N and C lobes of ncLf are sufficient to elicit anti-HCV effects in Huh7.5 cells. rcLf and its N lobe displayed similar HCV inhibitory effects to their native counterparts and may constitute an efficient and cost-effective approach for potential clinical applications.


Asunto(s)
Antivirales/metabolismo , Hepacivirus/efectos de los fármacos , Hepacivirus/patogenicidad , Hepatocitos/efectos de los fármacos , Hepatocitos/virología , Lactoferrina/genética , Lactoferrina/metabolismo , Animales , Productos Biológicos/metabolismo , Camelus , Línea Celular , Citoplasma/virología , Humanos , Reacción en Cadena de la Polimerasa , ARN Viral/análisis , ARN Viral/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Internalización del Virus/efectos de los fármacos
19.
J Proteome Res ; 10(12): 5409-15, 2011 Dec 02.
Artículo en Inglés | MEDLINE | ID: mdl-22084829

RESUMEN

Human milk contains many bioactive proteins that are likely to support the early development of the newborn. The aim of this study was to identify whether there are specific minor proteins associated with the human milk casein micelle prepared by the acid precipitation method. Protein identification was performed by liquid chromatography tandem mass spectrometry analysis. Eighty-two proteins were identified in the casein micelle, 18 of which are not present in their whey compartment. Thirty-two of these proteins specifically associated with the casein micelle have not previously been identified in human milk or colostrum. Proteins involved in immune function comprised the major part (28%) of total proteins, and another significant part is involved in metabolism/energy production (22%). Most of the proteins were of extracellular or cytoplasmic origin (accounting for 50 and 29%, respectively). This study indicates that various soluble proteins should be considered as part of the casein compartment, prepared by the acid precipitation method. The data provide new insight not only into the proteomic profile of the human milk casein micelle and its physiological significance, but also into the proper proportion of casein and casein-associated proteins to use in infant formula.


Asunto(s)
Caseínas/química , Leche Humana/química , Proteoma/análisis , Ácidos/química , Caseínas/análisis , Cromatografía Liquida , Calostro/química , Citoplasma/química , Bases de Datos de Proteínas , Metabolismo Energético , Femenino , Precipitación Fraccionada/métodos , Humanos , Micelas , Leche Humana/fisiología , Proteoma/química , Proteómica/métodos , Solubilidad , Espectrometría de Masas en Tándem
20.
J Proteome Res ; 10(8): 3530-41, 2011 Aug 05.
Artículo en Inglés | MEDLINE | ID: mdl-21714549

RESUMEN

The milk fat globule membrane (MFGM) contains proteins which have been implicated in a variety of health benefits. Milk fat globule membrane proteins were isolated from human milk during a 12 month lactation period and subjected to in-solution digestion and liquid chromatography tandem mass spectrometry analysis. Data were pooled, and our results showed that 191 proteins were identified. Relative quantification of the identified MFGM proteins during the course of lactation was performed by label free spectral counting and differentiation expression analysis, which showed some proteins decreasing during the course of lactation whereas some increased or remained at a relatively constant level. The human MFGM proteins are distributed between intracellular, extracellular, and membrane-associated proteins, and they are mainly involved in cell communication and signal transduction, immune function, metabolism and energy production. This study provides more insights into the dynamic composition of human MFGM proteins, which in turn will enhance our understanding of the physiological significance of MFGM proteins.


Asunto(s)
Glucolípidos/metabolismo , Glicoproteínas/metabolismo , Lactancia , Proteínas de la Membrana/metabolismo , Leche Humana/metabolismo , Proteómica , Western Blotting , Cromatografía Liquida , Femenino , Humanos , Gotas Lipídicas , Espectrometría de Masas en Tándem
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