RESUMO
OBJECTIVE@#Our aim was to explore whether heat stress protein (HSP) 9 preferentially expresses under heat stress and affects the expression of other heat stress proteins as well as to explore the effect of HSPB9 overexpression and knockdown on apoptosis in DF-1.@*METHODS@#We used gene cloning to construct an overexpression vector of the target gene, and synthesized the target gene interference fragment to transfect the chicken fibroblast cell line. Gene and protein expression, as well as apoptosis, were detected by RT-qPCR, Western blot, and flow cytometry.@*RESULTS@#Chicken DF-1 cells showed an early state of apoptosis in the early stages of HSPB9 overexpression. In the later stages, as HSPB9 expression increased, the cells showed inhibition of apoptosis. When the cells were under heat stress, HSPB9 expression was much higher and earlier than the expression of HSPB1 and HSPA2. In addition, high expression of HSPB9 had a negative effect on HSPB1 and HSPA2 expression. This negative feedback decreased the percentage of early stages of apoptotic cells and promoted cell survival.@*CONCLUSION@#HSPB9 expression, although rapid, is detrimental to cell survival early during its overexpression. In heat stress, HSPB9 overexpression, while inhibiting the expression of HSPA2 and HSPB1, is beneficial to cell survival.
Assuntos
Animais , Apoptose , Genética , Proteínas Aviárias , Genética , Linhagem Celular , Galinhas , Proteínas de Choque Térmico , Genética , Resposta ao Choque Térmico , GenéticaRESUMO
El síndrome de Wiskott-Aldrich es una inmunodeficiencia primaria; con una incidencia de 3,5 a 5,2 por cada millón de recién nacidos masculinos. Se caracteriza por tener un patrón de herencia recesiva ligada al cromosoma X. En estos pacientes; se ha descrito la tríada clásica de inmunodeficiencia; microtrombocitopenia y eczema. Presentamos un paciente de 5 años de edad; hispánico; con antecedentes de numerosas infecciones desde el primer año de vida. Actualmente; presenta desnutrición crónica; talla baja secundaria y retraso en el desarrollo del lenguaje. Se diagnosticó una mutación poco frecuente del gen asociado al síndrome de Wiskott-Aldrich.
The Wiskott-Aldrich syndrome is a rare X-linked recessive immunodeficiency, with an estimated incidence of 3.5 to 5.2 cases per million males. It is characterizedby immunodeficiency, microthrombocytopenia and eczema. We present a 5-year-old Hispanic male, with a medical history of numerous infectious diseases, compromised health, chronic malnutrition, language delay and failure to thrive. An infrequent mutation in the Wiskott-Aldrich syndrome gene was found.
Assuntos
Animais , Embrião de Galinha , Proteínas Aviárias/metabolismo , Caderinas/metabolismo , Células-Tronco Neurais/citologia , Células-Tronco Neurais/metabolismo , Proteínas Aviárias/antagonistas & inibidores , Proteínas Aviárias/genética , Sequência de Bases , Contagem de Células , Caderinas/antagonistas & inibidores , Caderinas/genética , Regulação da Expressão Gênica no Desenvolvimento , Técnicas de Silenciamento de Genes , Tubo Neural/citologia , Tubo Neural/embriologia , Tubo Neural/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Fenótipo , Interferência de RNA , RNA Interferente Pequeno/genética , Transdução de SinaisRESUMO
The bursa of Fabricius (BF) is the acknowledged central humoral immune organ in birds. Bursal septpeptide II (BSP-II) is an immunomodulatory bioactive peptide isolated from BF. To understand the effects of BSP-II on immune induction, gene expression profiles of hybridoma cells treated with BSP-II were evaluated. Pathway analysis showed that regulated genes were involved in cytokine-cytokine receptor interactions, T cell receptor signaling pathway, and pathway in cancer. It was observed that BSP-II reduced tumor cells proliferation and stimulated p53 expression. These results indicate potential mechanisms underlying the effects of the humoral immune system on immune induction, including antitumor activities. Our study has provided a novel insight into immunotherapeutic strategies for treating human tumors.
Assuntos
Animais , Antineoplásicos/farmacologia , Proteínas Aviárias/farmacologia , Bolsa de Fabricius/imunologia , Proliferação de Células/efeitos dos fármacos , Galinhas/imunologia , Hibridomas/efeitos dos fármacos , Fatores Imunológicos/farmacologia , Análise de Sequência com Séries de Oligonucleotídeos/veterinária , Transdução de Sinais/efeitos dos fármacos , TranscriptomaRESUMO
Characteristic clinical manifestations of Newcastle disease include leukopenia and immunosuppression. Peripheral blood mononuclear cells (PBMCs) are the main targets of Newcastle disease virus (NDV) infection. To survey changes in proteomic expression in chicken PBMCs following NDV infection, PBMC proteins from 30 chickens were separated using two-dimensional electrophoresis (2-DE) and subjected to mass spectrometry analysis. Quantitative intensity analysis showed that the expression of 78 proteins increased more than two-fold. Thirty-five proteins exhibited consistent changes in expression and 13 were identified as unique proteins by matrix assisted laser desorption ionization-time of flight mass spectrometer/mass spectrometer including three that were down-regulated and 10 that were up-regulated. These proteins were sorted into five groups based on function: macromolecular biosynthesis, cytoskeleton organization, metabolism, stress responses, and signal transduction. Furthermore, Western blot analysis confirmed the down-regulation of integrin-linked kinase expression and up-regulation of lamin A production. These data provide insight into the in vivo response of target cells to NDV infection at the molecular level. Additionally, results from this study have helped elucidate the molecular pathogenesis of NDV and may facilitate the development of new antiviral therapies as well as innovative diagnostic methods.
Assuntos
Animais , Proteínas Aviárias/genética , Galinhas , Regulação da Expressão Gênica , Leucócitos Mononucleares/enzimologia , Doença de Newcastle/genética , Vírus da Doença de Newcastle/fisiologia , Doenças das Aves Domésticas/genética , Proteoma , Organismos Livres de Patógenos Específicos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária , Espectrometria de Massas em Tandem/veterináriaRESUMO
The purpose of this study was to determine whether osteoprotegerin (OPG) could affect osteoclat differentiation and activation under serum-free conditions. Both duck embryo bone marrow cells and RAW264.7 cells were incubated with macrophage colony stimulatory factor (M-CSF) and receptor activator for nuclear factor kappaB ligand (RANKL) in serum-free medium to promote osteoclastogenesis. During cultivation, 0, 10, 20, 50, and 100 ng/mL OPG were added to various groups of cells. Osteoclast differentiation and activation were monitored via tartrate-resistant acid phosphatase (TRAP) staining, filamentous-actin rings analysis, and a bone resorption assay. Furthermore, the expression osteoclast-related genes, such as TRAP and receptor activator for nuclear factor kappaB (RANK), that was influenced by OPG in RAW264.7 cells was examined using real-time polymerase chain reaction. In summary, findings from the present study suggested that M-CSF with RANKL can promote osteoclast differentiation and activation, and enhance the expression of TRAP and RANK mRNA in osteoclasts. In contrast, OPG inhibited these activities under serum-free conditions.
Assuntos
Animais , Fosfatase Ácida/genética , Proteínas Aviárias/farmacologia , Células da Medula Óssea/efeitos dos fármacos , Células Cultivadas , Patos , Embrião não Mamífero/efeitos dos fármacos , Isoenzimas/genética , Fator Estimulador de Colônias de Macrófagos/metabolismo , Osteoclastos/citologia , Osteoprotegerina/farmacologia , Ligante RANK/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Receptor Ativador de Fator Nuclear kappa-B/genéticaRESUMO
The objective of the study was to clone avian beta-defensin (AvBD) 5 gene from pigeon bone marrow tissues and liver tissues, to express the recombinant AvBD5 protein in E. coli, and to determine its antimicrobial activity. The mRNA of duck AvBD5 was cloned from pigeon bone marrow tissues and liver tissues by RT-PCR. In addition, phylogenetic relationships between amino acid sequence of the pigeon AvBD5, AvBDs from other avian species, and some mammalian beta-defensin-5 were analyzed. The cDNA of pigeon AvBD5 was sub-cloned into pGEX-6p-1 vector to construct recombinant plasmid pGEX-pigeon AvBD5. The recombinant protein was expressed into E. coli and purified. Antimicrobial activity and physical-chemical stability of the recombinant fusion protein were measured in vitro. The complete nucleotide sequence of both cDNAs contained 201 bp nucleotides, encoding a polypeptide of 66 amino acids. Both beta-defensins have six conserved cysteines. Phylogenetic relationships were analyzed. Both pigeon AvBDs shared the highest amino acid homology (87.9% and 78.8%) with duck AvBD5. So it was named as pigeon AvBD5alpha (bone marrow) and AvBD5beta (liver). Both recombinant plasmids were transformed into E. coli BL21 and the bacteria were induced with Isopropyl beta-D-1-Thiogalactopyranoside (IPTG). After purification, antibacterial activity of the purified was investigated. In addition, effect of ionic strength on the antibacterial activity, and hemolytic recombinant protein activity of the purified recombinant protein were investigated. A 32 kDa protein was highly expressed. Both purified recombinant pigeon AvBD5alpha and AvBD5beta exhibited extensive antimicrobial activities against 12 bacteria, including Gram-positive and Gram-negative. In high salt ions concentrations, antibacterial activity of both recombinant proteins was decreased. In addition, the hemolysis activity of recombinant protein was extremely low.
Assuntos
Animais , Sequência de Aminoácidos , Anti-Infecciosos , Metabolismo , Farmacologia , Proteínas Aviárias , Genética , Farmacologia , Clonagem Molecular , Columbidae , Genética , Escherichia coli , Genética , Metabolismo , Dados de Sequência Molecular , Proteínas Recombinantes , Genética , Farmacologia , beta-Defensinas , Genética , FarmacologiaRESUMO
Sex determination is important for conservation and population studies, particularly for reproduction programs of threatened species and behavioural ecology. Turdus amaurochalinus, Creamy-bellied Thrush, only exhibits sexual dimorphism during the breeding season, when males are considered to show intense yellow bills, and females and immature males show dark brown bills. The objectives of this study were: 1) to determine the sex of individuals using genetic techniques, and 2) to test the hypothesis that sex dimorphism can be detected by morphometry. This study was carried out at Parque Nacional da Restinga de Jurubatiba, a preserved area located on the North coast of Rio de Janeiro State. The birds were captured using ornithological nets, singly marked with metal rings, weighed, measured and had blood samples collected before being released. The sex of 42 T. amaurochalinus individuals was determined using the CHD gene marker. A total of 20 males and 22 females were identified from June to August, with peak capture frequency in June. Turdus amaurochalinus females and males differed significantly in morphometrical measures. The most important traits to distinguish males from females were wing length (Student t-test=4.34, df=40, p=0.0001) and weight (Student t-test=2.08,df=40, p=0.044): females were heavier and had significantly shorter wing length than males. Females and males were correctly classified in 86% and 75% of cases, respectively, using Discriminant Analysis. The molecular analysis was the most secure method for sex determination in the studied species. Rev. Biol. Trop. 59 (2): 789- 794. Epub 2011 June 01.
La determinación del sexo es importante para la conservación y los estudios poblacionales. Turdus amaurochalinus no presenta aparente dimorfismo sexual. El objetivo de este estudio fue determinar el sexo a través de una técnica genética, mediante el uso del marcador del gen CHD y se puso a prueba la hipótesis de que el dimorfismo sexual puede ser detectado por morfometría. Este estudio se llevó a cabo en el Parque Nacional da Restinga de Jurubatiba, una zona protegida situada en la costa norte de Río de Janeiro. Las aves fueron capturadas con redes de niebla, los individuos se marcaron con anillos de metal, se pesaron, medieron y se les tomó una muestra de sangre antes de ser liberados. Un total de 20 machos y 22 hembras fueron identificados en el área de estudio desde junio hasta agosto, con la frecuencia máxima de captura en junio. La prueba de t-student fue usada para evaluar si hembras y machos se diferencian considerablemente en relación a medidas morfométricas. Los rasgos más importantes para distinguir machos de hembras fueron la longitud del ala y el peso: las hembras eran más pesadas y tenían longitud de ala considerablemente más corta que los machos. Hembras y machos fueron correctamente clasificados en un 86% y 75% de casos respectivamente, donde se usó un análisis discriminante. El análisis molecular es el método más seguro para la determinación sexual en la especie estudiada.
Assuntos
Animais , Feminino , Masculino , Proteínas Aviárias/genética , Proteínas de Ligação a DNA/genética , Passeriformes/genética , Análise para Determinação do Sexo/métodos , Marcadores Genéticos/genética , Passeriformes/anatomia & histologia , Passeriformes/classificaçãoRESUMO
Mouse models of cancer enable researchers to learn about tumor biology in complicated and dynamic physiological systems. Since the development of gene targeting in mice, cancer biologists have been among the most frequent users of transgenic mouse models, which have dramatically increased knowledge about how cancers form and grow. The Chinese Journal of Cancer will publish a series of papers reporting the use of mouse models in studying genetic events in cancer cases. This editorial is an overview of the development and applications of mouse models of cancer and directs the reader to upcoming papers describing the use of these models to be published in coming issues, beginning with three articles in the current issue.
Assuntos
Animais , Humanos , Camundongos , Vírus da Leucose Aviária , Genética , Proteínas Aviárias , Genética , Metabolismo , Modelos Animais de Doenças , Marcação de Genes , Camundongos Transgênicos , Metástase Neoplásica , Neoplasias Experimentais , Genética , Metabolismo , Interferência de RNA , Receptores Virais , Genética , MetabolismoRESUMO
The increasing incidence and mortality associated with advanced stages of melanoma are cause for concern. Few treatment options are available for advanced melanoma and the 5-year survival rate is less than 15%. Targeted therapies may revolutionize melanoma treatment by providing less toxic and more effective strategies. However, maximizing effectiveness requires further understanding of the molecular alterations that drive tumor formation, progression, and maintenance, as well as elucidating the mechanisms of resistance. Several different genetic alterations identified in human melanoma have been recapitulated in mice. This review outlines recent progress made in the development of mouse models of melanoma and summarizes what these findings reveal about the human disease. We begin with a discussion of traditional models and conclude with the recently developed RCAS/TVA somatic cell gene delivery mouse model of melanoma.
Assuntos
Animais , Humanos , Camundongos , 9,10-Dimetil-1,2-benzantraceno , Vírus da Leucose Aviária , Genética , Proteínas Aviárias , Genética , Metabolismo , Linhagem Celular Tumoral , Modelos Animais de Doenças , Técnicas de Transferência de Genes , Vetores Genéticos , Genética , Melanócitos , Metabolismo , Melanoma , Genética , Patologia , Melanoma Experimental , Genética , Camundongos Transgênicos , Transplante de Neoplasias , Receptores Virais , Genética , Metabolismo , Neoplasias Cutâneas , Genética , Patologia , Acetato de Tetradecanoilforbol , TransgenesRESUMO
The aggressive and invasive nature of brain tumors has hampered progress in the design and implementation of efficacious therapies. The recent success of targeted therapies in other tumor types makes this an attractive area for research yet complicating matters is the ability of brain tumors to circumvent the targeted pathways to develop drug resistance. Effective therapies will likely need to target more than one signaling pathway or target multiple nodes within a given pathway. Key to identifying these targets is the elucidation of the driver and passenger molecules within these pathways. Animal models provide a useful tool with many advantages in the study of these pathways. These models provide a means to dissect the critical components of tumorigenesis, as well as serve as agents for preclinical testing. This review focuses on the use of the RCAS/tv-a mouse model of brain tumors and describes their unique ability to provide insight into the role of oncogene cooperation in tumor development and progression.
Assuntos
Animais , Humanos , Camundongos , Vírus da Leucose Aviária , Genética , Proteínas Aviárias , Genética , Neoplasias Encefálicas , Genética , Patologia , Modelos Animais de Doenças , Progressão da Doença , Avaliação Pré-Clínica de Medicamentos , Métodos , Vetores Genéticos , Glioma , Genética , Patologia , Camundongos Transgênicos , Oncogenes , Genética , Receptores Virais , GenéticaRESUMO
A line of research beginning in the early 1960s with the observation that West Nile virus and, later, several strains of rabies virus could inhibit the development of the Rous sarcoma virus-induced tumor in the wing-web of chicken (a "sarcoma-blockade") eventually culminated in the characterization of a 14-kDa circulating anti-sarcoma and anti-viral activity christened "plasma factor" (PF) which, unlike the interferons, inhibited the replication of diverse RNA-containing viruses, but not of any DNA-containing viruses. The possibility that this 14 kDa protein represented a novel antiviral cytokine has been strengthened by analysis of partial amino acid sequencing data which suggest that this 14-kDa cytokine may correspond to the 127-amino acid-long chicken YB2-like protein (Locus: XP_423576) deduced very recently from the genomic sequencing of chicken. Biologically, proteins of the Y-box family (such as chicken YB1 and YB2) not only bind DNA and thus regulate transcription but also bind single-stranded RNA in a sequence-specific and reversible manner, repress viral RNA translation, inhibit retroviral transformation of chicken fibroblasts, and are known to regulate transcription of human immunodeficiency virus and hepatitis B virus. Taken together, the available data point to a novel anti-viral cytokine with a novel mechanism of action.
Assuntos
Sequência de Aminoácidos , Animais , Proteínas Aviárias/genética , Galinhas , Citocinas/fisiologia , Proteínas de Ligação a DNA/genética , Dados de Sequência Molecular , Vírus da Raiva/patogenicidade , Sarcoma Aviário/imunologia , Homologia de Sequência de Aminoácidos , Fatores de Transcrição/genética , Interferência Viral , Vírus do Nilo Ocidental/patogenicidadeRESUMO
<p><b>OBJECTIVE</b>To study the effects of constitutive nitric oxide synthase inhibitor L-nitroarginine on the recovery of traumatic facial paralysis in rats and the changes of the expression of cNOS and OX42 in the facial nucleus.</p><p><b>METHODS</b>L-nitroarginine was intraperitoneally injected into rats and the recovery of facial paralysis was observed at different time point. and the changes of cNOS and OX42 positive neurons were studied in facial nucleus.</p><p><b>RESULTS</b>Treatment of L-nitroarginine could remarkably inhibit the recovery of traumatic facial paralysis. The cNOS immunoactivity was obvious inhibited in facial nucleus, while the OX42 immunoactivity was obvious increased.</p><p><b>CONCLUSION</b>Endogenous nitric oxide may play an important mediator role on the recovery of traumatic facial paralysis.</p>