RESUMO
Understanding how bacteria evolve resistance to phages has implications for phage-based therapies and microbial evolution. In this study, the susceptibility of 335 Salmonella isolates to the wide host range Salmonella phage BPSELC-1 was tested. Potentially significant gene sets that could confer resistance were identified using bioinformatics approaches based on phage susceptibility phenotypes; more than 90 potential antiphage defense gene sets, including those involved in lipopolysaccharide (LPS) biosynthesis, DNA replication, secretion systems, and respiratory chain, were found. The evolutionary dynamics of Salmonella resistance to phage were assessed through laboratory evolution experiments, which showed that phage-resistant mutants rapidly developed and exhibited genetic heterogeneity. Most representative Salmonella hosts (58.1% of 62) rapidly developed phage resistance within 24 h. All phage-resistant mutant clones exhibited genetic heterogeneity and observed mutations in LPS-related genes (rfaJ and rfaK) as well as other genes such as cellular respiration, transport, and cell replication-related genes. The study also identified potential trade-offs, indicating that bacteria tend to escape fitness trade-offs through multi-site mutations, all tested mutants increased sensitivity to polymyxin B, but this does not always affect their relative fitness or biofilm-forming capacity. Furthermore, complementing the rfaJ mutant gene could partially restore the phage sensitivity of phage-resistant mutants. These results provide insight into the phage resistance mechanisms of Salmonella and the complexity of bacterial evolution resulting from phage predation, which can inform future strategies for phage-based therapies and microbial evolution.
Assuntos
Bacteriófagos , Bacteriófagos/genética , Lipopolissacarídeos , Salmonella , Mutação , Fenótipo , BactériasRESUMO
Bone health problems are a serious threat to laying hens; microbiome-based therapies, which are harmless and inexpensive, may be an effective solution for bone health problems. Here, we examined the impacts of supplementation with Clostridium butyricum (CB) on bone and immune homeostasis in pullets. The results of in vivo experiments showed that feeding the pullets CB was beneficial to the development of the tibia and upregulated the levels of the bone formation marker alkaline phosphatase and the marker gene runt-related transcription factor 2 (RUNX2). For the immune system, CB treatment significantly upregulated IL-10 expression and significantly increased the proportion of T regulatory (Treg) cells in the spleen and peripheral blood lymphocytes. In the in vitro test, adding CB culture supernatant or butyrate to the osteoblast culture system showed no significant effects on osteoblast bone formation, while adding lymphocyte culture supernatant significantly promoted bone formation. In addition, culture supernatants supplemented with treated lymphocytes (pretreated with CB culture supernatants) stimulated higher levels of bone formation. In sum, the addition of CB improved bone health by modulating cytokine expression and the ratio of Treg cells in the immune systems of layer pullets. Additionally, in vitro CB could promote the bone formation of laying hen osteoblasts through the mediation of lymphocytes.
Assuntos
Galinhas , Clostridium butyricum , Animais , Feminino , Galinhas/metabolismo , Suplementos Nutricionais , Desenvolvimento Ósseo , Citocinas/metabolismoRESUMO
Genetic diversity provides a rich repository for understanding the role of proteostasis in the management of the protein fold in human biology. Failure in proteostasis can trigger multiple disease states, affecting both human health and lifespan. Niemann-Pick C1 (NPC1) disease is a rare genetic disorder triggered by mutations in NPC1, a multi-spanning transmembrane protein that is trafficked through the exocytic pathway to late endosomes (LE) and lysosomes (Ly) (LE/Ly) to globally manage cholesterol homeostasis. Defects triggered by >300 NPC1 variants found in the human population inhibit export of NPC1 protein from the endoplasmic reticulum (ER) and/or function in downstream LE/Ly, leading to cholesterol accumulation and onset of neurodegeneration in childhood. We now show that the allosteric inhibitor JG98, that targets the cytosolic Hsp70 chaperone/co-chaperone complex, can significantly improve the trafficking and post-ER protein level of diverse NPC1 variants. Using a new approach to model genetic diversity in human disease, referred to as variation spatial profiling, we show quantitatively how JG98 alters the Hsp70 chaperone/co-chaperone system to adjust the spatial covariance (SCV) tolerance and set-points on an amino acid residue-by-residue basis in NPC1 to differentially regulate variant trafficking, stability, and cholesterol homeostasis, results consistent with the role of BCL2-associated athanogene family co-chaperones in managing the folding status of NPC1 variants. We propose that targeting the cytosolic Hsp70 system by allosteric regulation of its chaperone/co-chaperone based client relationships can be used to adjust the SCV tolerance of proteostasis buffering capacity to provide an approach to mitigate systemic and neurological disease in the NPC1 population.
Assuntos
Variação Genética/fisiologia , Proteínas de Choque Térmico HSP70/metabolismo , Proteína C1 de Niemann-Pick/metabolismo , Doença de Niemann-Pick Tipo C/genética , Regulação Alostérica/genética , Regulação Alostérica/fisiologia , Colesterol/metabolismo , Citosol/metabolismo , Retículo Endoplasmático/metabolismo , Endossomos/metabolismo , Variação Genética/genética , Proteínas de Choque Térmico HSP70/genética , Humanos , Lisossomos/metabolismo , Proteína C1 de Niemann-Pick/genéticaRESUMO
Effective phage cocktails consisting of multiple virus types are essential for successful phage therapy against pandrug-resistant pathogens, including Salmonella enterica serovar (S.) Typhimurium. Here we show that a Salmonella phage, F118P13, with non-productive infection and a lytic phage, PLL1, combined to inhibit pandrug-resistant S. Typhimurium growth and significantly limited resistance to phages in vitro. Further, intraperitoneal injection with this unique phage combination completely protected mice from Salmonella-induced death and inhibited bacterial proliferation rapidly in various organs. Furthermore, the phage combination treatment significantly attenuated the inflammatory response, restored the generation of CD4+ T cells repressed by Salmonella, and allowed macrophages and granulocytes to participate in immunophage synergy to promote bacterial clearance. Crucially, the non-productive phage F118P13 is less likely to be cleared by the immune system in vivo, thus providing an alternative to phage cocktail against bacterial infections.
Assuntos
Bacteriófagos , Fagos de Salmonella , Salmonella enterica , Camundongos , Animais , Salmonella typhimurium , Sistema ImunitárioRESUMO
Klebsiella pneumoniae is an opportunistic pathogen commonly associated with nosocomial infections. In our previous study, we have demonstrated that colistin-resistant K. pneumoniae is more susceptible to killing by lytic tailed phages than the colistin-sensitive parent strain, including T1-like ФNJS1. This fitness cost associated with colistin resistance is due to the alteration of the surface charge that promotes phage adherence and infection. However, the receptor for phage adsorption has not been identified. In this study, we found that ФNJS1 specifically infected nonmucoid K. pneumoniae isolates, and the accelerated phage adsorption to colistin-resistant nonmucoid K. pneumoniae cells is reversible. Further research suggested that bacteria lipopolysaccharide may be involved in phage reversible adsorption, while capsule polysaccharide may block the receptors on cell surface from phage attachment. Transposon mutagenesis of colistin-resistant K. pneumoniae revealed that mutation in wecA and wecG, two genes involved in lipopolysaccharide O-antigen biosynthesis, significantly deceased phage adsorption capacity and infection efficiency. Inactivation of wzyE, which leaded to the shorten of O-antigen chain length, enhanced phage infectivity. Moreover, mutation of the outer membrane protein FepA slowed the phage lysis rate, suggesting that FepA may be an irreversible receptor for ФNJS1. In summary, our results show a delicate balance between ФNJS1 and its hosts, where the lipopolysaccharide O-antigen may serve as an essential reversible receptor for phage NJS1, while the long O-antigen chain hinders the bacteriophage infection.
Assuntos
Bacteriófagos , Infecções por Klebsiella , Bacteriófagos/genética , Colistina , Humanos , Klebsiella pneumoniae , Mutagênese , Antígenos ORESUMO
In this study, we developed a real-time quantitative polymerase chain reaction (qPCR) assay based on a dual-labeled hydrolysis probe to simultaneously detect both duck circovirus (DuCV) 1 and DuCV-2. The reproducibility, sensitivity and specificity of the primer set and probe were evaluated using other duck pathogens. The detection limit was 20 copies per µL. The intra-assay coefficients of variation (CVs) were ≤ 0.73% and the inter-assay CVs were ≤ 1.89%. No cross-reaction occurred with other duck pathogens. In addition, the qPCR assay was successfully applied to the simultaneous detection of DuCV-1 and DuCV-2 in clinical field samples. Therefore, this assay will be useful for laboratory diagnosis and epidemiological field studies of DuCV.
Assuntos
Infecções por Circoviridae/diagnóstico , Circovirus/isolamento & purificação , Reação em Cadeia da Polimerase , Doenças das Aves Domésticas/diagnóstico , Animais , Infecções por Circoviridae/genética , Infecções por Circoviridae/virologia , Circovirus/genética , Circovirus/patogenicidade , DNA Viral/genética , Genótipo , Hidrólise , Doenças das Aves Domésticas/genética , Doenças das Aves Domésticas/virologia , Reação em Cadeia da Polimerase em Tempo RealRESUMO
To access the nutritional quality of the Ruditapes philippinarum, a comprehensive quality evaluation procedure is always important to be established. In this study, fifteen nutritional quality evaluation indicators of R. philippinarum from 7 months were analyzed, and the most important indicators were determined using a combination of multiple chemometric methods such as correlation analysis (CA), principal component analysis (PCA), and system cluster analysis (SCA). Significant differences in nutritional quality were observed across the 7 months, as per the ANOVA results (P < 0.05). The coefficient of variation values for the fifteen evaluation indicators for R. philippinarum across 7 months was 1.67-43.47%. The CA results revealed that some indicators were correlated to each other within a certain range. Four principal components with eigen-values > 1 were obtained with PCA, and a cumulative contribution of 92.11% was achieved. In addition, four essential quality indicators were extracted using SCA. Using these four indicators, a simple and efficient procedure can be applied for quality control in aquaculture.
RESUMO
Long noncoding RNAs (lncRNAs) have been shown to have critical regulatory roles in tumorigenesis. lncRNA LINC01561 (LINC01561) is a newly identified tumor-related lncRNA and its dysregulation has been demonstrated in several tumors. However, whether LINC01561 is involved in the progression of non-small-cell lung carcinoma (NSCLC) and its underlying mechanisms remain unknown. In this study, we first provided evidence that LINC01561 expressions were distinctly upregulated in NSCLC tissues and cell lines. Combining with bioinformatics assays and mechanism experiments, our group demonstrated that LINC01561 was activated by SOX2 in NSCLC. Clinical research revealed that upregulation of LINC01561 was related to poorer clinicopathologic features and shorter survival time. Functionally, suppression of LINC01561 exhibited tumor-suppressive functions through impairing cell proliferation, migration, and invasion as well as inducing apoptosis. Moreover, we verified that LINC01561 could directly bind to miR-760, isolating miR-760 from its target gene SHC SH2 domain-binding protein 1 (SHCBP1). We also found that SHCBP1 was lowly expressed in NSCLC and served as a tumor promoter. A functional study indicated that LINC01561 regulated SHCBP1 expression by competitively binding to miR-760. In summary, our findings indicated that SOX2-induced overexpression of LINC01561 promoted the proliferation and metastasis by acting as a competing endogenous RNA to modulate SHCBP1 by sponging miR-760.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Neoplasias Pulmonares/genética , MicroRNAs/genética , RNA Longo não Codificante/genética , Fatores de Transcrição SOXB1/genética , Proteínas Adaptadoras da Sinalização Shc/genética , Regulação para Cima/genética , Células A549 , Apoptose/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Progressão da Doença , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Neoplasias Pulmonares/patologia , Invasividade Neoplásica/genética , Invasividade Neoplásica/patologia , Transdução de Sinais/genéticaRESUMO
BACKGROUND: Salmonella has been considered as one of the most important foodborne pathogens that threatened breeding industry and public health. To investigate the prevalence and characterization of Salmonella isolated from duck farms and a slaughterhouse in Shandong province, a total of 49 Salmonella strains were isolated from 2342 samples from four duck farms and one duck slaughterhouse in Jinan and Tai'an, Shandong province, China. RESULTS: Among the isolates, S. Enteritidis (20/49, 40.8%) and S. Anatum (10/49, 20.4%) were the most prevalent, and high resistance rates were detected for erythromycin (49/49, 100.0%) and nalidixic acid (47/49, 95.9%). Class I integrons were detected in 17 isolates (34.7%17/49), which contained gene cassettes aadA7 + aac3-Id(15/17) and aadA5 + dfrA17 (2/17). Eleven different kinds of resistance genes were detected while blaTEM(36/49, 73.5%) was the most prevalent, followed by sul2(14/49, 28.6%). Thirteen virulence genes were tested, and all of the strains carried invA, hilA and sipA. Multilocus sequence typing (MLST) results showed that seven sequence types (STs) were identified; ST11 was the most prevalent ST (20/49, 40.8%), followed by ST2441 (10/49, 20.4%). There was a strong correlation between STs and serovars. The results of pulsed field gel electrophoresis(PFGE) showed that 39 PFGE patterns were generated from 49 Salmonella strains. PFGE patterns were mostly diverse and revealed high similarity between the isolates from the same sampling sites. CONCLUSIONS: The presence of Salmonella infections among duck farms revealed that ducks could also be potential reservoirs for Salmonella. The high resistance rates against commonly used antimicrobials suggested a need for more reasonable use of antimicrobials, as well as for investigating substitutes for antimicrobials.
Assuntos
Antibacterianos/farmacologia , Farmacorresistência Bacteriana Múltipla , Doenças das Aves Domésticas/microbiologia , Salmonelose Animal/microbiologia , Salmonella/isolamento & purificação , Matadouros/estatística & dados numéricos , Animais , China , Patos , Fazendas/estatística & dados numéricos , Integrons , Testes de Sensibilidade Microbiana , Tipagem de Sequências Multilocus , Salmonella/classificação , Salmonella/efeitos dos fármacos , Salmonella/genéticaRESUMO
Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) is a polyhydroxyalkanoate (PHA) with broad application prospects in various biomedical fields. Our previous study demonstrated the great biocompatibility of PHBHHx with neural stem cells (NSCs) in vitro. In this study, we went a step further and implanted the NSC-carrying PHBHHx film to in vivo into the lesion region in a rat traumatic brain injury (TBI) model. The in vivo biocompatibility of PHBHHx, as well as the survival, migration and differentiation of the transplanted NSCs were investigated. The results showed that PHBHHx did not induce additional reactive gliosis and supported the in vivo survival and growth of the transplanted stem cells. The transplanted NSCs were able to migrate into the brain tissue and differentiated into both neurons and astrocytes. Moreover, PHBHHx seemed to be able to maintain the stemness of the attached NSCs both in vitro and in vivo. The major monomer of PHBHHx, 3-hydroxybutyrate (3-HB), might contribute to this bioactivity because the addition of low concentrations of 3-HB facilitated the self-renewal of NSCs by shortening the G1 phase, promoted their proliferation, and enhanced the expression of a key regulatory transcription factor which helped to keep the stemness of NSCs. These results highlight the application of PHBHHx as a promising bioactive material for NSC transplantation in the brain and open up a wide space for further studies on functional recovery following the transplantation of NSCs attached to PHBHHx.
Assuntos
Lesões Encefálicas Traumáticas/terapia , Diferenciação Celular/efeitos dos fármacos , Células-Tronco Neurais/transplante , Neurônios/efeitos dos fármacos , Ácido 3-Hidroxibutírico/química , Animais , Astrócitos/química , Caproatos/química , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Modelos Animais de Doenças , Humanos , Células-Tronco Neurais/efeitos dos fármacos , Ratos , Transplante de Células-Tronco/métodosRESUMO
Autotaxin (ATX), which is highly expressed and secreted by adipocytes, functions as the key enzyme to generate lysophosphatidic acid (LPA) from lysophosphatidylcholine. Adipose tissue is the main source of circulating ATX that modulates plasma LPA levels. Upregulation of ATX expression in obese patients and mice is closely related with insulin resistance and impaired glucose tolerance. However, the mechanism of ATX expression in adipocytes remains largely unknown. In this study, we found that glycoprotein 130 (gp130)-mediated Janus kinase (JAK)-signal transducer and activator of transcription 3 (STAT3) activation was required for abundant ATX expression in adipocytes. Through gp130, the interleukin 6 (IL-6) family cytokines, such as IL-6, leukemia inhibitory factor, cardiotrophin-1, and ciliary neurotrophic factor, upregulated ATX expression in adipocytes. ATX contributes to the induction of insulin resistance and lipolysis in IL-6-stimulated adipocytes. Oral administration of gp130 inhibitor SC144 suppressed ATX expression in adipose tissue, decreased plasma ATX, LPA, and FFA levels, and significantly improved insulin sensitivity and glucose tolerance in high-fat diet-fed obese mice. In summary, our results indicate that the activation of gp130-JAK-STAT3 pathway by IL-6 family cytokines has an important role in regulating ATX expression in adipocytes and that gp130 is a promising target in the management of obesity-associated glucose metabolic diseases.
Assuntos
Adipócitos/efeitos dos fármacos , Receptor gp130 de Citocina/metabolismo , Regulação para Baixo/efeitos dos fármacos , Resistência à Insulina , Obesidade/patologia , Diester Fosfórico Hidrolases/metabolismo , Transdução de Sinais/efeitos dos fármacos , Células 3T3-L1 , Adipócitos/metabolismo , Adipócitos/patologia , Animais , Dieta/efeitos adversos , Ácidos Graxos não Esterificados/sangue , Glucose/metabolismo , Hidrazinas/farmacologia , Interleucina-6/metabolismo , Isoxazóis/farmacologia , Janus Quinases/metabolismo , Lipólise/efeitos dos fármacos , Lisofosfolipídeos/sangue , Camundongos , Obesidade/induzido quimicamente , Obesidade/metabolismo , Propionatos/farmacologia , Quinoxalinas/farmacologia , Fator de Transcrição STAT3/metabolismoRESUMO
The aim of current study was to investigate the metabolic changes associated with histidine supplementation in serum and urine metabolic signatures and serum amino acid (AA) profiles. Serum and urine 1H NMR-based metabolomics and serum AA profiles were employed in 32 and 37 obese women with metabolic syndrome (MetS) intervened with placebo or histidine for 12 weeks. Multivariable statistical analysis were conducted to define characteristic metabolites. In serum 1H NMR metabolic profiles, increases in histidine, glutamine, aspartate, glycine, choline, and trimethylamine-N-oxide (TMAO) were observed; meanwhile, decreases in cholesterol, triglycerides, fatty acids and unsaturated lipids, acetone, and α/ß-glucose were exhibited after histidine supplement. In urine 1H NMR metabolic profiles, citrate, creatinine/creatine, methylguanidine, and betaine + TMAO were higher, while hippurate was lower in histidine supplement group. In serum AA profiles, 10 AAs changed after histidine supplementation, including increased histidine, glycine, alanine, lysine, asparagine, and tyrosine and decreased leucine, isoleucine, ornithine, and citrulline. The study showed a systemic metabolic response in serum and urine metabolomics and AA profiles to histidine supplementation, showing significantly changed metabolism in AAs, lipid, and glucose in obese women with MetS.
Assuntos
Histidina/farmacologia , Metaboloma/efeitos dos fármacos , Adulto , Aminoácidos/sangue , Aminoácidos/efeitos dos fármacos , Aminoácidos/metabolismo , Glicemia/efeitos dos fármacos , Glicemia/metabolismo , Suplementos Nutricionais , Feminino , Histidina/administração & dosagem , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Síndrome Metabólica/tratamento farmacológico , Síndrome Metabólica/metabolismo , Pessoa de Meia-Idade , Obesidade/metabolismo , Espectroscopia de Prótons por Ressonância Magnética , Soro/química , Urina/químicaRESUMO
Autotaxin (ATX) is a key enzyme that converts lysophosphatidylcholine (LPC) into lysophosphatidic acid (LPA), a lysophospholipid mediator that regulates cellular activities through its specific G protein-coupled receptors. The ATX-LPA axis plays an important role in various physiological and pathological processes, especially in inflammation and cancer development. Although the transcriptional regulation of ATX has been widely studied, the post-transcriptional regulation of ATX is largely unknown. In this study, we identified conserved adenylate-uridylate (AU)-rich elements in the ATX mRNA 3'-untranslated region (3'UTR). The RNA-binding proteins HuR and AUF1 directly bound to the ATX mRNA 3'UTR and had antagonistic functions in ATX expression. HuR enhanced ATX expression by increasing ATX mRNA stability, whereas AUF1 suppressed ATX expression by promoting ATX mRNA decay. HuR and AUF1 were involved in ATX regulation in Colo320 human colon cancer cells and the LPS-stimulated human monocytic THP-1 cells. HuR knockdown suppressed ATX expression in B16 mouse melanoma cells, leading to inhibition of cell migration. This effect was reversed by AUF1 knockdown to recover ATX expression or by the addition of LPA. These results suggest that the post-transcriptional regulation of ATX expression by HuR and AUF1 modulates cancer cell migration. In summary, we identified HuR and AUF1 as novel post-transcriptional regulators of ATX expression, thereby elucidating a novel mechanism regulating the ATX-LPA axis.
Assuntos
Regiões 3' não Traduzidas , Movimento Celular , Proteína Semelhante a ELAV 1/metabolismo , Regulação Neoplásica da Expressão Gênica , Ribonucleoproteínas Nucleares Heterogêneas Grupo D/metabolismo , Proteínas de Neoplasias/metabolismo , Diester Fosfórico Hidrolases/biossíntese , Estabilidade de RNA , RNA Neoplásico/metabolismo , Animais , Proteína Semelhante a ELAV 1/genética , Técnicas de Silenciamento de Genes , Células HeLa , Ribonucleoproteína Nuclear Heterogênea D0 , Ribonucleoproteínas Nucleares Heterogêneas Grupo D/genética , Humanos , Camundongos , Proteínas de Neoplasias/genética , Diester Fosfórico Hidrolases/genética , RNA Neoplásico/genéticaRESUMO
Aspirin (ASA) is a cardioprotective drug with anti-cardiac fibrosis action in vivo. This study was aimed to clarify the anti-cardiac fibrosis action of ASA and the underlying mechanisms. Two heart injury models (injection of isoproterenol and ligation of the left anterior descending branch) were used in mice to induce cardiac fibrosis. The animals were treated with ASA (10 mg·kg-1·d-1, ig) for 21 and 14 d, respectively. ASA administration significantly improved cardiac function, and ameliorated heart damage and fibrosis in the mice. The mechanisms underlying ASA's anti-fibrotic effect were further analyzed in neonatal cardiac fibroblasts (CFs) exposed to hypoxia in vitro. ASA (0.5-5 mmol/L) dose-dependently inhibited the proliferation and Akt phosphorylation in the CFs. In addition, ASA significantly inhibited CF apoptosis, and decreased the levels of apoptosis markers (cleaved caspase 3 and Parp1), which might serve as a side effect of anti-fibrotic effect of ASA. Furthermore, ASA dose-dependently inhibited the autophagy in the CFs, as evidenced by the reduced levels of autophagy marker LC3-II. The autophagy inhibitor Pepstatin A (PepA) promoted the inhibitory effect of ASA on CF proliferation, whereas the autophagy inducer rapamycin rescued ASA-caused inhibition of CF proliferation, suggesting an autophagy-dependent anti-proliferative effect of ASA. Both p38 inhibitor SB203580 and ROS scavenger N-acetyl-cysteine (NAC) significantly decreased Akt phosphorylation in CFs in the basal and hypoxic situations, but they both significantly increased LC3-II levels in the CFs. Our results suggest that an autophagy- and p38/ROS-dependent pathway mediates the anti-cardiac fibrosis effect of ASA in CFs. As PepA and SB203580 did not affect ASA-caused inhibition of CF apoptosis, the drug combination will enhance ASA's therapeutic effects.
Assuntos
Aspirina/uso terapêutico , Autofagia/efeitos dos fármacos , Cardiomiopatias/tratamento farmacológico , Cardiotônicos/uso terapêutico , Animais , Apoptose/efeitos dos fármacos , Aspirina/farmacologia , Cardiomiopatias/patologia , Cardiomiopatias/fisiopatologia , Cardiotônicos/farmacologia , Hipóxia Celular , Fibroblastos/efeitos dos fármacos , Fibroblastos/patologia , Fibrose/tratamento farmacológico , Fibrose/patologia , Imidazóis/farmacologia , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Piridinas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidoresRESUMO
BACKGROUND: Lin28a enhances glucose uptake and insulin-sensitivity. However, the role of Lin28a on experimental diabetic cardiomyopathy (DCM) is not well understood. We investigated the potential role and mechanism ofLin28a in diabetes-induced myocardial dysfunction in mice. METHODS: Diabetes was induced by intraperitoneal (i.p.) injections of Streptozocin (STZ) in mice. Animals were randomized to be treated with lentivirus carrying Lin28a siRNA or Lin28a cDNA. Cardiac function, cardiomyocyte autophagy, apoptosis and mitochondria morphology in diabetic mice were compared between groups. The target proteins of Lin28a were examined by western blot analysis. RESULTS: Lin28a levels were markedly reduced in the cardiac tissue compared to the control mice. Lin28a overexpression significantly improved left ventricular ejection fraction (LVEF), promoted autophagy, decreased myocardial apoptotic index and alleviated mitochondria cristae destruction in diabetic mice. Lin28a knockdown exacerbated diabetic injury as evidenced by decreased LVEF, increased apoptotic index and aggravated mitochondria cristae destruction. Interestingly, pretreatment with a PKA inhibitor, N-[2-(p-Bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide], di-HCl Salt (H89) abolished the beneficial effects of Lin28a overexpression. RhoA-expression and ROCK2-expression were decreased in vivo after Lin28a overexpression, while Lin28a knockdown increased the expression of RhoA and ROCK2 in diabetic mice. CONCLUSIONS: Lin28a protects against DCM through PKA/ROCK2 dependent pathway. Lin28a might serve as a potential therapeutic target for the treatment of the patients with DCM.
Assuntos
Cardiomiopatias Diabéticas/complicações , Cardiomiopatias Diabéticas/fisiopatologia , Proteínas de Ligação a RNA/metabolismo , Disfunção Ventricular Esquerda/etiologia , Disfunção Ventricular Esquerda/fisiopatologia , Quinases Associadas a rho/metabolismo , Animais , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Transdução de SinaisRESUMO
BACKGROUND: The prevalence of food allergy has been increasing, but treatment is very limited. DNA vaccination has been recognized as a promising method for the treatment of allergic diseases; however, poor immunogenicity has hindered its application. METHODS: BALB/c mice were intradermally injected with plasmid DNA encoding the peanut protein Ara h 2 (pAra h 2) or pAra h 2 pretreated with poly-L- lysine (PLL) before or after sensitization with Ara h 2 protein. Ara h 2-specific antibodies were measured by ELISA. CD207+ dendritic cells (DCs) and Treg cells in draining lymph nodes were analyzed by flow cytometry after DNA immunization, and cytokine production in splenocytes was also analyzed. RESULTS: In the prophylactic study, pretreatment with pAra h 2 or PLL-pAra h 2 resulted in lower levels of Ara h 2-specific IgG1, IgG2a, and IgE after sensitization with Ara h 2 protein, and mice in the PLL-pAra h 2 group had a significantly lower level of antibodies than those in the pAra h 2 group. In the treatment study, intradermal injection with pAra h 2 or PLL-pAra h 2 after Ara h 2 protein sensitization significantly decreased the level of Ara h 2-specific antibodies, and PLL- pAra h 2 had stronger effects than pAra h 2. There were increased numbers of CD207+ DCs and Treg cells in the mice receiving intradermal injection with PLL-pAra h 2, and splenocytes from PLL-pAra h 2-treated mice secreted increased levels of IFN-γ and IL-10. CONCLUSIONS: Modification of pAra h 2 with PLL improved its prophylactic and therapeutic effects in peanut-allergic mice.
Assuntos
Albuminas 2S de Plantas/imunologia , Antígenos de Plantas/imunologia , Arachis/efeitos adversos , Glicoproteínas/imunologia , Hipersensibilidade a Amendoim/imunologia , Hipersensibilidade a Amendoim/prevenção & controle , Vacinas de DNA/imunologia , Albuminas 2S de Plantas/química , Albuminas 2S de Plantas/genética , Animais , Antígenos de Plantas/química , Antígenos de Plantas/genética , Movimento Celular/imunologia , Citocinas/metabolismo , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Modelos Animais de Doenças , Feminino , Glicoproteínas/química , Glicoproteínas/genética , Imunização , Imunoglobulina E/sangue , Imunoglobulina E/imunologia , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Linfonodos/imunologia , Camundongos , Polilisina/química , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo , Vacinas de DNA/química , Vacinas de DNA/genéticaRESUMO
Marek's disease virus (MDV), the aetiological agent of Mareks' disease (MD), is a highly cell-associated oncogenic α-herpesvirus that replicates in chicken lymphocytes and establishes a latent infection within CD4(+) T cells. We investigated the possible effect of MDV infection on the exacerbation of necrotic dermatitis in the combs of MD-susceptible (72) and MD-resistant (63) chicken lines at 21 days post infection. MDV-infected birds of line 63 are relatively resistant to tumour development but exhibit an unusual necrosis of combs, wattles, and footpads that is intensified when infected with MDV. Chickens from line 72, on the other hand, are highly susceptible to MDV infection and tumour development. Real-Time PCR analysis revealed that IL-6, IL-8, IL-12, IL-18, iNOS, and IFNγ were all up regulated in the comb tissues of MDV-infected susceptible line 72 with no visible necrotic damage. With the exception of IL-8 and iNOS, the expression of all the other tested genes was barely detected in the necrotic combs of the resistant line 63. Real-Time PCR analysis revealed the MDV meq oncogene transcripts in the spleen tissues of both infected lines but in the comb tissues of only the susceptible line 72. A significant infiltration of macrophages and lymphocytes was detected in the comb tissues of both resistant and susceptible lines. Histopathological analysis also showed thinning and erosion of epidermis and inflammation, lympho-plasmocytic infiltration, heterophilic, and histocytic cellulitis within the connective tissues of the necrotic combs. Gram stain of the sectioned frozen comb samples exposed the presence of Gram-positive micrococcus.
Assuntos
Galinhas/virologia , Dermatoses do Pé/veterinária , Herpesvirus Galináceo 2/imunologia , Doença de Marek/virologia , Doenças das Aves Domésticas/virologia , Animais , Biomarcadores/análise , Galinhas/genética , Galinhas/imunologia , Dermatoses do Pé/imunologia , Dermatoses do Pé/patologia , Dermatoses do Pé/virologia , Perfilação da Expressão Gênica/veterinária , Herpesvirus Galináceo 2/genética , Herpesvirus Galináceo 2/isolamento & purificação , Inflamação/veterinária , Doença de Marek/imunologia , Doença de Marek/patologia , Necrose/veterinária , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/patologia , Baço/patologia , Baço/virologiaRESUMO
BACKGROUND: Phosphatidylcholine (PC), the major source of dietary choline, has been demonstrated to improve the capability of learning and memory in rodent and the amelioration of long-chain n-3 polyunsaturated fatty acids (PUFA) on anti-aging and anti-oxidation is widely known as well. In this study, three kinds of PC were chose to demonstrate the role of different fatty acids composition on glycerol backbone in improving the brain function of mice induced by scopolamine which was used to impair cholinergic system and cause oxidative stress. METHODS: Male BALB/c mice were randomly divided into 5 groups: model (M) group, control (Con) group, egg yolk lecithin (EL) group, squid PC (SQ-PC) group and sea cucumber PC (SC-PC) group. The intraperitoneal injection of scopolamine hydrobromide (5 mg/kg) was carried out on the 8(th) of group feeding and sustained daily until the end of test. Morris water maze test was used to evaluate the improvement of cognitive decline and the activity of acetylcholinesterase (AchE), superoxide dismutase (SOD) and monoamine oxidase (MAO) and malondialdehyde (MDA) content in brain were measured to assess the physiological changes. RESULTS: In behavior test, the latency of PC groups was significantly reduced, while number of crossing the platform and time in target quadrant were increased in comparison with M group and the improvements of SQ-PC and SC-PC were better than that of EL (P < 0.05). Similar trend was observed in physiological changes. The AchE activity was effectively decreased and the SOD activity increased in hippocampus, cortex and white matter when comparing PC groups with M group. SQ-PC, SC-PC and EL respectively showed 22.82, 28.80 and 11.81 % decrease in MDA level in brain compared with M group. The MAO activity in white matter of SQ-PC, SC-PC and EL group separately depressed 33.05, 33.64 and 19.73 % in comparison with M group. No significance between SQ-PC and SC-PC was found in these indicators except the SOD activity in hippocampus and white matter. SQ-PC group had a higher SOD activity in hippocampus (103.68U/mg · prot.) and lower in white matter (120.57 U/mg · prot.) than SC-PC group (95.53 U/mg · prot. in hippocampus, 134.49 U/mg · prot. in white matter). PC rich in n-3 PUFA acted more ameliorative effects than that barely contained on the indicators above. CONCLUSIONS: Different fatty acids composition of PC all could diminish the cognitive decline and biological damage and protect the brain. EPA and DHA partly enhaced to the advantageous effects.
Assuntos
Encéfalo/efeitos dos fármacos , Demência/dietoterapia , Fosfatidilcolinas/química , Fosfatidilcolinas/farmacologia , Escopolamina/toxicidade , Acetilcolinesterase/metabolismo , Animais , Encéfalo/metabolismo , Decapodiformes/química , Demência/induzido quimicamente , Modelos Animais de Doenças , Gema de Ovo/química , Ácidos Graxos Ômega-3/química , Ácidos Graxos Ômega-3/farmacologia , Masculino , Aprendizagem em Labirinto/efeitos dos fármacos , Camundongos Endogâmicos BALB C , Monoaminoxidase/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Pepinos-do-Mar/químicaRESUMO
Non-structural protein 1 (NS1) binds small interfering RNA and suppresses RNA silencing in plants, but the underlying mechanism of this suppression is not well understood. Therefore, here we characterized NS1 encoded by the avian influenza virus H9N2. The NS1 protein was able to suppress RNA silencing induced by either sense RNA or double-stranded RNA (dsRNA). Using deletion and point mutants, we discovered that the first 70 residues of NS1 could suppress RNA silencing triggered by sense transgene, but this sequence was not sufficient to block dsRNA-induced silencing. Any mutations of two arginine residues (35R and 46R) of NS1, which contribute to its homodimeric structure, caused the loss of its silencing suppression activity. These results indicate that the region after residue 70 of NS1 is essential for the repression activity on dsRNA-induced RNA silencing, and that the dimeric structure of NS1 plays a critical role in its RNA silencing suppression function.
Assuntos
Regulação da Expressão Gênica de Plantas , Interações Hospedeiro-Patógeno , Vírus da Influenza A Subtipo H9N2/imunologia , Vírus da Influenza A Subtipo H9N2/fisiologia , Interferência de RNA , Proteínas não Estruturais Virais/metabolismo , Animais , Galinhas , Análise Mutacional de DNA , Regulação da Expressão Gênica , Vírus da Influenza A Subtipo H9N2/genética , Influenza Aviária/virologia , Proteínas Mutantes/genética , Proteínas Mutantes/metabolismo , Mutação Puntual , RNA Interferente Pequeno/metabolismo , Deleção de Sequência , Nicotiana/genética , Nicotiana/metabolismo , Proteínas não Estruturais Virais/genéticaRESUMO
Genetic variation in human populations can result in the misfolding and aggregation of proteins, giving rise to systemic and neurodegenerative diseases that require management by proteostasis. Here, we define the role of GRP94, the endoplasmic reticulum Hsp90 chaperone paralog, in managing alpha-1-antitrypsin deficiency on a residue-by-residue basis using Gaussian process regression-based machine learning to profile the spatial covariance relationships that dictate protein folding arising from sequence variants in the population. Covariance analysis suggests a role for the ATPase activity of GRP94 in controlling the N- to C-terminal cooperative folding of alpha-1-antitrypsin responsible for the correction of liver aggregation and lung-disease phenotypes of alpha-1-antitrypsin deficiency. Gaussian process-based spatial covariance profiling provides a standard model built on covariant principles to evaluate the role of proteostasis components in guiding information flow from genome to proteome in response to genetic variation, potentially allowing us to intervene in the onset and progression of complex multi-system human diseases.