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1.
Nano Lett ; 24(8): 2544-2552, 2024 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-38349341

RESUMO

Labeling the genome and envelope of a virus with multicolor quantum dots (QDs) simultaneously enables real-time monitoring of viral uncoating and genome release, contributing to our understanding of virus infection mechanisms. However, current labeling techniques require genetic modification, which alters the virus's composition and infectivity. To address this, we utilized the CRISPR/Cas13 system and a bioorthogonal metabolic method to label the Japanese encephalitis virus (JEV) genome and envelopes with different-colored QDs in situ. This technique allows one-step two-color labeling of the viral envelope and intraviral genome with QDs harnessing virus infection. In combination with single-virus tracking, we visualized JEV uncoating and genome release in real time near the endoplasmic reticulum of live cells. This labeling strategy allows for real-time visualization of uncoating and genome release at the single-virus level, and it is expected to advance the study of other viral infection mechanisms.


Assuntos
Pontos Quânticos , Viroses , Vírus , Humanos , Envelope Viral/metabolismo , Proteínas do Envelope Viral
2.
Fungal Genet Biol ; 173: 103899, 2024 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-38802054

RESUMO

Fusarium head blight is a devastating disease that causes severe yield loses and mycotoxin contamination in wheat grain. Additionally, balancing the trade-off between wheat production and disease resistance has proved challenging. This study aimed to expand the genetic tools of the endophyte Phomopsis liquidambaris against Fusarium graminearum. Specifically, we engineered a UDP-glucosyltransferase-expressing P. liquidambaris strain (PL-UGT) using ADE1 as a selection marker and obtained a deletion mutant using an inducible promoter that drives Cas9 expression. Our PL-UGT strain converted deoxynivalenol (DON) into DON-3-G in vitro at a rate of 71.4 % after 36 h. DON inactivation can be used to confer tolerance in planta. Wheat seedlings inoculated with endophytic strain PL-UGT showed improved growth compared with those inoculated with wildtype P. liquidambaris. Strain PL-UGT inhibited the growth of Fusarium graminearum and reduced infection rate to 15.7 %. Consistent with this finding, DON levels in wheat grains decreased from 14.25 to 0.56 µg/g when the flowers were pre-inoculated with PL-UGT and then infected with F. graminearum. The expression of UGT in P. liquidambaris was nontoxic and did not inhibit plant growth. Endophytes do not enter the seeds nor induce plant disease, thereby representing a novel approach to fungal disease control.

3.
Mar Drugs ; 22(2)2024 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-38393065

RESUMO

It is important to improve the production of bioactive secondary products for drug development. The Escherichia coli-Streptomyces shuttle vector pSET152 and its derived vector pIB139 containing a strong constitutive promoter ermEp* are commonly used as integrative vectors in actinomycetes. Four new integrative vectors carrying the strong constitutive promoter kasOp*, hrdBp, SCO5768p, and SP44, respectively, were constructed and proven to be functional in different mangrove-derived Streptomyces host strains by using kanamycin resistance gene neo as a reporter. Some biosynthetic genes of elaiophylins, azalomycin Fs, and armeniaspirols were selected and inserted into these vectors to overexpress in their producers including Streptomyces sp. 219807, Streptomyces sp. 211726, and S. armeniacus DSM 43125, resulting in an approximately 1.1-1.4-fold enhancement of the antibiotic yields.


Assuntos
Actinobacteria , Streptomyces , Streptomyces/genética , Antibacterianos , Regiões Promotoras Genéticas/genética , Vetores Genéticos , Actinobacteria/genética , Plasmídeos
4.
Int Wound J ; 21(4): e14509, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38151959

RESUMO

This meta-analysis investigates the impact of psychological interventions on the wound healing process at surgical sites in patients with psoriasis who have undergone various surgical procedures. Following the PRISMA guidelines, an extensive database search was conducted, initially identifying 679 articles, with 6 studies ultimately meeting our rigorous selection criteria. These studies, which included both Randomized Controlled Trials and observational designs, utilized a range of scales, such as the REEDA and Manchester Scar Scale (MSS), to measure the healing of surgical wounds. Statistical analyses were performed using Review Manager and SPSS, revealing that psychological interventions significantly expedited wound healing as early as 1 week post-surgery (I2 = 93%; Random: SMD = -3.01, 95% CI: [-4.35, -1.66], p < 0.01), according to the REEDA scale. At the one-month follow-up, a continued positive effect was observed on the MSS (I2 = 69%; Random: SMD = 2.31, 95% CI: [1.54, 3.08], p < 0.01). The studies demonstrated a low risk of bias, and funnel plot analysis suggested no significant publication bias. These results highlight the beneficial role of psychological support in the postoperative recovery of psoriasis patients, suggesting a need for a more integrated approach to patient care that includes psychological well-being as a component of comprehensive treatment strategies.


Assuntos
Intervenção Psicossocial , Ferida Cirúrgica , Humanos , Infecção da Ferida Cirúrgica , Cicatrização , Cicatriz
5.
Angew Chem Int Ed Engl ; : e202402880, 2024 May 17.
Artigo em Inglês | MEDLINE | ID: mdl-38758629

RESUMO

Lysine-specific peptide and protein modification strategies are widely used to study charge-related functions and applications. However, these strategies often result in the loss of the positive charge on lysine, significantly impacting the charge-related properties of proteins. Herein, we report a strategy to preserve the positive charge and selectively convert amines in lysine side chains to amidines using nitriles and hydroxylamine under aqueous conditions. Various unprotected peptides and proteins were successfully modified with a high conversion rate. Moreover, the reactive amidine moiety and derived modification site enable subsequent secondary modifications. Notably, positive charges were retained during the modification. Therefore, positive charge-related protein properties, such as liquid-liquid phase separation behaviour of α-synuclein, were not affected. This strategy was subsequently applied to a lysine rich protein to develop an amidine-containing coacervate DNA complex with outstanding mechanical properties. Overall, our innovative strategy provides a new avenue to explore the characteristics of positively charged proteins.

6.
Bioconjug Chem ; 34(6): 1037-1044, 2023 06 21.
Artigo em Inglês | MEDLINE | ID: mdl-37204067

RESUMO

Sphingomyelinase (SMase), a hydrolase of sphingomyelin (SM) enriched in the outer leaflet of the plasma membrane of mammalian cells, is closely associated with the onset and development of many diseases, but the specific mechanisms of SMase on the cell structure, function, and behavior are not yet fully understood due to the complexity of the cell structure. Artificial cells are minimal biological systems constructed from various molecular components designed to mimic cellular processes, behaviors, and structures, which are excellent models for studying biochemical reactions and dynamic changes in cell membranes. In this work, we presented an artificial cell model that mimics the lipid composition and content of the outer leaflet of mammalian plasma membranes for studying the effect of SMase on cell behavior. The results confirmed that the artificial cells can respond to SM degradation by producing ceramides that enrich and alter the membrane charge and permeability, thus inducing the budding and fission of the artificial cells. Thus, the artificial cells developed here provide a powerful tool to study the mechanism of action of cell membrane lipids on cell biological behavior, paving the way for further molecular mechanism studies.


Assuntos
Células Artificiais , Esfingomielinas , Animais , Esfingomielinas/análise , Esfingomielinas/metabolismo , Esfingomielinas/farmacologia , Ceramidas/química , Ceramidas/metabolismo , Ceramidas/farmacologia , Membrana Celular/metabolismo , Esfingomielina Fosfodiesterase/química , Esfingomielina Fosfodiesterase/metabolismo , Esfingomielina Fosfodiesterase/farmacologia , Mamíferos/metabolismo
7.
BMC Cancer ; 23(1): 1126, 2023 Nov 18.
Artigo em Inglês | MEDLINE | ID: mdl-37980466

RESUMO

BACKGROUND: Shared decision-making is useful to facilitate cancer treatment decisions. However, it is difficult to make treatment decisions when physician and patient preferences are different. This review aimed to summarize and compare the preferences for cancer treatments between physicians and patients. METHODS: A systematic literature search was conducted on PubMed, Embase, PsycINFO, CINAHL and Scopus. Studies elicited and compared preferences for cancer treatments between physicians and patients were included. Information about the study design and preference measuring attributes or questions were extracted. The available relative rank of every attribute in discrete choice experiment (DCE) studies and answers to preference measuring questions in non-DCE studies were summarized followed by a narrative synthesis to reflect the preference differences. RESULTS: Of 12,959 studies identified, 8290 were included in the title and abstract screening and 48 were included in the full text screening. Included 37 studies measured the preferences from six treatment-related aspects: health benefit, adverse effects, treatment process, cost, impact on quality of life, and provider qualification. The trade-off between health benefit and adverse effects was the main focus of the included studies. DCE studies showed patients gave a higher rank on health benefit and treatment process, while physicians gave a higher rank on adverse effects. Non-DCE studies suggested that patients were willing to take a higher risk of adverse effects or lower health benefit than physicians when accepting a treatment. CONCLUSIONS: Physicians and patients had important preference differences for cancer treatment. More sufficient communication is needed in cancer treatment decision-making.


Assuntos
Neoplasias , Médicos , Humanos , Comportamento de Escolha , Preferência do Paciente , Qualidade de Vida , Neoplasias/terapia
8.
J Pept Sci ; 29(3): e3454, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36181422

RESUMO

Considering the fact that site-selective late-stage diversification of peptides and proteins remains a challenge for biochemistry, strategies targeting low-abundance natural amino acids need to be further developed. As an extremely oxidation-sensitive and low-abundance amino acid, methionine emerges as a promising target for chemo- and site-selective modification. Herein we report an efficient and highly selective modification on methionine residues by one-pot O- and N-transfer reaction, generating sulfoximine-modified peptides with near-perfect conversion within 10 min. Moreover, the great tolerance to other natural amino acids has been demonstrated in reactions with various peptide substrates. To demonstrate the generality of this protocol, we have modified natural peptides and obtained sulfoximination products with high conversion rates. This methodology provides a novel strategy as the expansion of the methionine-based peptide functionalization toolbox.


Assuntos
Metionina , Proteínas , Metionina/química , Metionina/metabolismo , Proteínas/química , Peptídeos/química , Racemetionina/metabolismo , Estresse Oxidativo
9.
Int J Mol Sci ; 25(1)2023 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-38203450

RESUMO

Methyl farnesoate (MF), a crucial sesquiterpenoid hormone, plays a pivotal role in the reproduction of female crustaceans, particularly in the vitellogenesis process. Despite extensive research on its functions, the molecular mechanisms that regulate MF levels during the vitellogenic phase remain largely elusive. This study investigates the roles of microRNAs (miRNAs), significant post-transcriptional regulators of gene expression, in controlling MF levels in the swimming crab Portunus trituberculatus. Through bioinformatic analysis, four miRNAs were identified as potential regulators targeting two genes encoding Carboxylesterases (CXEs), which are key enzymes in MF degradation. Dual luciferase reporter assays revealed that let-7b and miR-141 suppress CXE1 and CXE2 expression by directly binding to their 3' UTRs. In vivo overexpression of let-7b and miR-141 significantly diminished CXE1 and CXE2 levels, consequently elevating hemolymph MF and enhancing vitellogenin expression. Spatiotemporal expression profile analysis showed that these two miRNAs and their targets exhibited generally opposite patterns during ovarian development. These findings demonstrate that let-7b and miR-141 collaboratively modulate MF levels by targeting CXEs, thus influencing vitellogenesis in P. trituberculatus. Additionally, we found that the expression of let-7b and miR-141 were suppressed by MF, constituting a regulatory loop for the regulation of MF levels. The findings contribute novel insights into miRNA-mediated ovarian development regulation in crustaceans and offer valuable information for developing innovative reproduction manipulation techniques for P. trituberculatus.


Assuntos
Braquiúros , Ácidos Graxos Insaturados , MicroRNAs , Vitelogênese , Animais , Feminino , Regiões 3' não Traduzidas , Braquiúros/genética , Hidrolases de Éster Carboxílico , MicroRNAs/genética , Vitelogênese/genética
10.
Int J Mol Sci ; 24(8)2023 Apr 21.
Artigo em Inglês | MEDLINE | ID: mdl-37108789

RESUMO

The Chinese tongue sole (Cynoglossus semilaevis) is a traditional, precious fish in China. Due to the large growth difference between males and females, the investigation of their sex determination and differentiation mechanisms receives a great deal of attention. Forkhead Box O (FoxO) plays versatile roles in the regulation of sex differentiation and reproduction. Our recent transcriptomic analysis has shown that foxo genes may participate in the male differentiation and spermatogenesis of Chinese tongue sole. In this study, six Csfoxo members (Csfoxo1a, Csfoxo3a, Csfoxo3b, Csfoxo4, Csfoxo6-like, and Csfoxo1a-like) were identified. Phylogenetic analysis indicated that these six members were clustered into four groups corresponding to their denomination. The expression patterns of the gonads at different developmental stages were further analyzed. All members showed high levels of expression in the early stages (before 6 months post-hatching), and this expression was male-biased. In addition, promoter analysis found that the addition of C/EBPα and c-Jun transcription factors enhanced the transcriptional activities of Csfoxo1a, Csfoxo3a, Csfoxo3b, and Csfoxo4. The siRNA-mediated knockdown of the Csfoxo1a, Csfoxo3a, and Csfoxo3b genes in the testicular cell line of Chinese tongue sole affected the expression of genes related to sex differentiation and spermatogenesis. These results have broadened the understanding of foxo's function and provide valuable data for studying the male differentiation of tongue sole.


Assuntos
Linguados , Linguado , Animais , Feminino , Masculino , Filogenia , Linguados/genética , Linguados/metabolismo , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Sequência de Aminoácidos , Testículo/metabolismo , Linguado/genética
11.
Anal Chem ; 94(21): 7567-7575, 2022 05 31.
Artigo em Inglês | MEDLINE | ID: mdl-35581735

RESUMO

Mammalian cell display technology uses eukaryotic protein expression system to display proteins on cell surfaces and has become an important method in biological research. Although mammalian cell display technology has many advantages and development potential, certain attributes of the displayed protein remain uncharacterized, such as whether the displayed proteins re-enter the cell and how displayed proteins move into the cell. Here, we present the endocytosis mechanism, motility behavior, and transport kinetics of displayed proteins determined using HaloTag as the displayed protein and quantum dot-based single-particle tracking. The displayed protein enters the cell through clathrin-mediated endocytosis and is transported through the cell in three stages, which is dependent on microfilaments and microtubules. The dynamic information obtained in this study provides answers to questions about endocytosis and postendocytosis transport of displayed proteins and, therefore, is expected to facilitate the development of surface display technology.


Assuntos
Pontos Quânticos , Citoesqueleto de Actina , Animais , Membrana Celular , Endocitose , Mamíferos
12.
Fungal Genet Biol ; 158: 103650, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-34923123

RESUMO

Fusarium head blight (FHB) is a disease that affects wheat crops worldwide and is caused by Fusarium graminearum. Effective and safe strategies for the prevention and treatment of the disease are very limited. Phomopsis liquidambaris, a universal endophyte, can colonize wheat. Two engineered strains, Phomopsis liquidambaris OE-Chi and IN-Chi, were constructed by transformation with a plasmid and integration of a chitinase into the genome, respectively. The OE-Chi and IN-Chi strains could inhibit the expansion of Fusarium sp. in plate confrontation assays in vitro. Colonization of the OE-Chi strain in wheat showed better effects than colonization of the IN-Chi strain and alleviated the inhibition of wheat growth caused by F. graminearum. The shoot length, root length and fresh weight of infected wheat increased by 164.9%, 115.4%, and 190.7%, respectively, when the plants were inoculated with the OE-Chi strain. The peroxidase (POD) activity in the wheat root increased by 38.0%, and it was maintained at a high level in the shoot, which suggested that the OE-Chi strain could enhance the resistance of wheat to F. graminearum. The root and shoot superoxide dismutase (SOD) activities were decreased by 11.8% and 19.0%, respectively, which may be helpful for colonization by the OE-Chi strain. These results suggested that the Phomopsis liquidambaris OE-Chi strain may be a potential endophyte in the biocontrol of FHB.


Assuntos
Quitinases , Fusarium , Ascomicetos , Quitinases/genética , Endófitos/genética , Fusarium/genética , Doenças das Plantas , Triticum
13.
Value Health ; 25(10): 1768-1777, 2022 10.
Artigo em Inglês | MEDLINE | ID: mdl-35710892

RESUMO

OBJECTIVES: This study aimed to compare the treatment preference among oncologists, patients with lung cancer, and their family members in China. METHODS: A face-to-face discrete choice experiment survey was conducted among oncologists, patients, and their family members. Participants completed 13 choice sets describing 6 key attributes, namely, overall survival time, risk of severe adverse effect, severity of pain, appetite, physical functioning status, and monthly cost. Mixed logit model and latent class analysis were used to estimate attribute level preference weights and the relative importance (RI) for attributes. The willingness to pay (WTP) and maximum acceptable risk (MAR) were also estimated. The RI, WTP, and MAR of oncologists, patients, and family members were compared. RESULTS: A total of 121 oncologists and 161 dyads of patients and family members completed the survey. Overall survival time, physical functioning status, and pain were the 3 most important attributes across all 3 groups. Oncologists and family members had higher RI on overall survival time than patients (48% and 51% vs 38%). Patients had higher RI on physical functioning status and pain (23% and 14%) than oncologists (13% and 12%) and family members (16% and 11%). For extending survival, patients had the least WTP, and family members had the highest MAR. The latent class analysis identified 2 classes in the patient group and 3 classes in oncologist and family member groups. CONCLUSIONS: There were differences in preferences for survival, risk, quality of life, and costs associated with cancer treatments among patients, oncologists, and family members. This finding highlights the need of involving patients in treatment decision making in China.


Assuntos
Neoplasias , Oncologistas , China , Comportamento de Escolha , Família , Humanos , Neoplasias/terapia , Dor , Preferência do Paciente , Qualidade de Vida , Inquéritos e Questionários
14.
Fish Physiol Biochem ; 48(5): 1365-1375, 2022 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-36125598

RESUMO

The phenomenon of sexual size dimorphism (SSD), existing in mammals, birds, reptiles, spiders, amphibians, insects, and fishes, is generally related to feeding efficiency, energy allocation, sex steroids, and somatotropic and reproductive endocrine axes. Recently, positive and negative regulations of sex steroids have been reported on SSD in various species. Chinese tongue soles (Cynoglossus semilaevis) at 4 months were fed with 17ß-estradiol (E2) and testosterone (T) supplemented feeds for 8 months to assess the effect of sex steroids on growth traits in different sexes. The potential genetic regulation was examined using several growth-related genes. The results showed that two sex steroid hormones had inhibitory effects on the growth performance of different sexes of C. semilaevis. At the age of 8 months, the expression of insulin-like growth factor 2 gene (igf2), 24-dehydrocholesterol reductase (dhcr24), leptin, and estrogen receptor 2 (esr2) in the liver showed an overall downward trend. The expression of insulin-like growth factor 1 (igf1) was reduced, while thyroid hormone receptor-associated protein 3 (thrap3) expression tended to increase in the gonad after T and E2 treatments. In the brain, somatostatin 1, tandem duplicate 2 (sst1.2) expression increased with the treatment of T and E2 (P < 0.05), while growth hormone-releasing hormone (ghrh) expression decreased. E2 and T had different effects on growth differentiation factor 8 (gdf8) and insulin-like growth factor-binding protein 7 (igfbp7) expression in the muscle. Expression of gdf8 increased in the treated fishes in contrast to the reduction expression of igfbp7. This study provided important clues for understanding the role of sex steroids in flatfish SSD.


Assuntos
Linguados , Linguado , Animais , Estradiol/metabolismo , Testosterona/metabolismo , Fator de Crescimento Insulin-Like I/genética , Leptina/metabolismo , Miostatina/metabolismo , Receptor beta de Estrogênio/metabolismo , Desmosterol/metabolismo , Hormônios Esteroides Gonadais/metabolismo , Linguado/metabolismo , Hormônio Liberador de Hormônio do Crescimento , Peixes/metabolismo , Língua/metabolismo , Somatostatina , Receptores dos Hormônios Tireóideos , Oxirredutases/metabolismo , Linguados/genética , Mamíferos/metabolismo
15.
Anal Chem ; 92(12): 8614-8622, 2020 06 16.
Artigo em Inglês | MEDLINE | ID: mdl-32452205

RESUMO

In this study, we proposed a ratiometric electrochemiluminescent (ECL)/electrochemical (EC) biosensor based on duplex-specific nuclease (DSN)-assisted target recycling and multilayer catalytic hairpin assembly (CHA) amplification cascades for the detection of microRNA (miRNA). The DSN-assisted target recycling transformed miRNAs into a large number of ssDNA, which then catalyzed a multilayer CHA amplification cascade to produce numerous long dsDNA duplexes Hn/Hn+1 (n = 2, 4, 6, ...). Then the Hn/Hn+1 displaced the ferrocene (Fc)-labeled ssDNA (Sx+1, x = 1, 3, 5, ...) to hybridize with the Sx sequence on the gold electrode surface. Consequently, a great number of long Sx/Hn/Hn+1 duplexes were immobilized for binding Ru(phen)32+ to obtain an amplified ECL signal. Meanwhile, the EC signal of Fc was reduced, and the quenching effect of Fc to ECL signal also decreased. By measuring the ratio of the ECL signal of Ru(phen)32+ to the EC signal of Fc, quantitative analysis of miRNA-499 with high accuracy and reproducibility was obtained. The ratiometric biosensor shows high sensitivity and a wide linear range of 6 orders of magnitude. With the help of DSN-assisted target recycling, this strategy can be easily extended to detect other miRNAs without redesigning the CHA cascade system. The proposed "hybrid" ratiometric ECL/EC strategy enriches the ratiometric sensors and can find extensive applications in bioanalysis, especially for multiplex detection.


Assuntos
Técnicas Eletroquímicas , Compostos Ferrosos/química , Medições Luminescentes , Metalocenos/química , MicroRNAs/sangue , Catálise , DNA/química , Humanos , MicroRNAs/química
16.
Int J Syst Evol Microbiol ; 69(3): 828-832, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30663961

RESUMO

A bacterial strain, designated HHTR 118T, was isolated from a culture of the green alga Ulvaprolifera obtained from offshore seawater of Qingdao, Shandong Province, China. Cells of strain HHTR 118T were rod-shaped and motile with a single flagellum, and approximately 0.3-0.4 µm wide and 0.8-1.4 µm long. The strain was Gram-stain-negative, strictly aerobic, catalase-negative and oxidase-positive. Optimal growth was observed at 30 °C, at pH 8.0 and with 1 % (w/v) NaCl. Nitrate was not reduced. Sucrose, sodium citrate and l-leucine stimulated growth, but not lactose, fructose, xylose, d-mannose, glucose, raffinose, rhamnose, ornithine or lysine. The DNA G+C content of strain HHTR 118T calculated on the basis of the genome sequence was 64.9 mol% and the genome size is 4.6 Mbp. The major quinone was ubiquinone 10 and the predominant cellular fatty acids (>10 % of total fatty acids) were summed feature 8 (C18 : 1ω6c and/or C18 : 1ω7c). The predominant polar lipids were phosphatidylglycerol, one unidentified phospholipid, two unidentified aminolipids and three unidentified polar lipids. Phylogenetic analysis, based on 16S rRNA gene sequences, demonstrated that strain HHTR 118T was affiliated with the family Rhodospirillaceae. On the basis of the 16S rRNA gene sequence data as well as physiological and biochemical characteristics, we concluded that strain HHTR 118T represents a novel species of a novel genus. We propose the name of Algihabitans albus gen. nov., sp. nov. for this novel species. The type strain of the novel species is strain HHTR 118T (=KCTC 62395T=MCCC 1K03486T).


Assuntos
Filogenia , Rhodospirillaceae/classificação , Ulva/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Rhodospirillaceae/isolamento & purificação , Água do Mar/microbiologia , Análise de Sequência de DNA , Ubiquinona/química
17.
Am J Physiol Cell Physiol ; 311(4): C572-C582, 2016 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-27488664

RESUMO

Oxidative stress and impaired antioxidant defense are believed to be contributors to the cardiovascular aging process. The transcription factor nuclear factor-E2-related factor 2 (Nrf2) plays a key role in orchestrating cellular antioxidant defenses and maintaining redox homeostasis. Our previous study showed that Exendin-4, a glucagon-like peptide-1 analog, alleviates angiotensin II (ANG II)-induced vascular smooth muscle cell (VSMC) senescence by inhibiting Rac1 activation via cAMP/PKA (Zhao L, Li AQ, Zhou TF, Zhang MQ, Qin XM. Am J Physiol Cell Physiol 307: C1130-C1141, 2014). The objective of this study is to investigate if Nrf2 mediates the antisenescent effect of Exendin-4 in ANG II-induced VSMCs. Here we report that Exendin-4 triggered Nrf2 nuclear translocation, a downstream target of cAMP-responsive element-binding protein (CREB) and expressions of antioxidant genes heme oxygenase-1 (HO-1) and NAD(P)H quinone oxidoreductase-1 (NQO-1) in a dose- and time-dependent manner. In addition, knock-down of Nrf2 attenuated the inhibitory effects of Exendin-4 on ANG II-induced superoxidant generation and VSMC senescence. PKA/CREB pathway participated in the upregulations of HO-1 and NQO-1 induced by Exendin-4. Notably, our study revealed that Exendin-4 dose-dependently increased the acetylation of Nrf2 and the recruitment of transcriptional coactivator CREB binding protein (CBP) to Nrf2. The Exendin-4-induced Nrf2 transactivation was diminished in the presence of CBP small interfering RNA. Microscope imaging of Nrf2, as well as immunoblotting for Nrf2, showed that the Exendin-4-evoked Nrf2 acetylation favored its nuclear retention. Importantly, CBP silencing attenuated the suppressing effects of Exendin-4 on ANG II-induced VSMC senescence and superoxidant production. In conclusion, these results provide a mechanistic insight into how Nrf2 signaling mediates the antisenescent and antioxidative effects induced by Exendin-4 in VSMCs.


Assuntos
Angiotensina II/efeitos adversos , Senescência Celular/efeitos dos fármacos , Músculo Liso Vascular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Fator 2 Relacionado a NF-E2/metabolismo , Peptídeos/farmacologia , Substâncias Protetoras/farmacologia , Peçonhas/farmacologia , Animais , Antioxidantes/metabolismo , Células Cultivadas , Exenatida , Peptídeo 1 Semelhante ao Glucagon/metabolismo , Heme Oxigenase-1/metabolismo , Masculino , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , NADH NADPH Oxirredutases/metabolismo , Oxirredução , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/fisiologia , Ratos , Ratos Sprague-Dawley , Transdução de Sinais/efeitos dos fármacos
18.
Mycopathologia ; 181(5-6): 329-39, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26700222

RESUMO

Candida albicans is an opportunistic human pathogen, and its pathogenicity is associated with hyphal formation. Previous studies have shown that at neutral-to-alkaline pH, hyphal growth is dependent on the Rim101 pathway whose activation requires Snf7, a member of the ESCRT system. In this work, we described the purification and characterization of the C. albicans Vps4, an AAA ATPase required for recycling of the ESCRTs. Its role on hyphal growth has been investigated. Our data suggest deletion of Vps4 decreases overall hyphal growth at pH 7 and increases the growth of multiple hyphae induced by serum, which indicates that the ESCRTs may make a Rim101-independent contribution to hyphal growth. Furthermore, DBeQ, an inhibitor of the AAA ATPase p97, was shown to inhibit the ATPase activity of Vps4 with an IC50 of about 11.5 µM. To a less degree, it also inhibits hyphal growth. Our work may provide a new strategy to control C. albicans infection.


Assuntos
Adenosina Trifosfatases/metabolismo , Candida albicans/enzimologia , Candida albicans/crescimento & desenvolvimento , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Inibidores Enzimáticos/metabolismo , Proteínas Fúngicas/metabolismo , Hifas/crescimento & desenvolvimento , Quinazolinas/metabolismo , Fatores de Virulência/metabolismo , Adenosina Trifosfatases/genética , Adenosina Trifosfatases/isolamento & purificação , Complexos Endossomais de Distribuição Requeridos para Transporte/genética , Complexos Endossomais de Distribuição Requeridos para Transporte/isolamento & purificação , Proteínas Fúngicas/genética , Proteínas Fúngicas/isolamento & purificação , Deleção de Genes , Humanos , Concentração de Íons de Hidrogênio , Concentração Inibidora 50 , Fatores de Virulência/genética , Fatores de Virulência/isolamento & purificação
19.
Appl Opt ; 54(20): 6186-9, 2015 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-26193392

RESUMO

A simple novel method for the absolute interferometric testing of spherical surfaces is presented. This approach yields an estimate for test surface errors without changing experimental settings, such as cavity length, which may affect the apparent reference errors. The test surface is tested in three orientations: a basic position and two rotation positions. Full-surface absolute maps for each test piece are determined with the proposition that any arbitrary wavefront of a circular cross section may be expressed as a linear combination of polynomial terms and a data-processing technique based on differencing rotation maps of the rotated surface. An optimized numerical reconstruction algorithm employing the least-squares technique to determine the true azimuthal positions of part rotation is used to reconstruct the rotational data. The technique does not require any assumptions about the surfaces under test and additional measurements. The differencing rotation method provides a new approach for rotationally asymmetric removal and azimuthal errors correction in the absolute test. Experimental results are presented to compare the method proposed to the classic two-sphere method.

20.
Artigo em Inglês | MEDLINE | ID: mdl-38788625

RESUMO

Ovarian development in animals is a complicated biological process, requiring the simultaneous coordination among various genes and pathways. To understand the dynamic changes and molecular regulatory mechanisms of ovarian development in mud crab (Scylla paramamosain), both histological observation and whole transcriptome sequencing of ovarian tissues at different mating stages were implemented in this study. The histological results revealed that ovarian development was delayed in unmated females (60 days after courtship behavior but not mating), who exhibited an oocyte diameter of 56.38 ± 15.17 µm. Conversely, mated females exhibited accelerated the ovarian maturation process, with females reaching ovarian stage III (proliferative stage) 23 days after mating and attained an average oocyte diameter of 132.19 ± 15.07 µm. Thus, mating process is essential in promoting the rapid ovarian development in mud crab. Based on the whole transcriptome sequencing analysis, a total of 518 mRNAs, 1502 lncRNAs, 18 circRNAs and 151 miRNAs were identified to be differentially expressed between ovarian tissues at different mating stages. Notably, six differentially expressed genes (DEGs) associated with ovarian development were identified, including ovary development-related protein, red pigment concentrating hormone receptor, G2/mitotic-specific cyclin-B3-like, lutropin-chorio gonadotropic hormone receptor, renin receptor, and SoxB2. More importantly, both DEGs and targets of differentially expressed non-coding RNAs (DEncRNAs) were enriched in renin-angiotensin system, TGF-ß signaling, cell adhesion molecules, MAPK signaling pathway, and ECM-receptor interaction, suggesting that these pathways may play significant roles in the ovarian development of mud crabs. Moreover, competition endogenous RNA (ceRNA) networks were constructed while mRNAs were differentially expressed between mating stages were involved in Gene Ontology (GO) biological processes such as developmental process, reproduction, and growth. These findings could provide solid foundations for the future development of female mud crab maturation enhancement strategy, and improve the understanding of the ovarian maturation process in crustaceans.


Assuntos
Braquiúros , Ovário , Transcriptoma , Animais , Feminino , Braquiúros/genética , Braquiúros/crescimento & desenvolvimento , Ovário/crescimento & desenvolvimento , Ovário/metabolismo , Comportamento Sexual Animal , Perfilação da Expressão Gênica , Masculino , Análise de Sequência de RNA
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