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1.
Chem Rev ; 124(1): 124-163, 2024 01 10.
Artigo em Inglês | MEDLINE | ID: mdl-38033123

RESUMO

Psychedelics make up a group of psychoactive compounds that induce hallucinogenic effects by activating the serotonin 2A receptor (5-HT2AR). Clinical trials have demonstrated the traditional psychedelic substances like psilocybin as a class of rapid-acting and long-lasting antidepressants. However, there is a pressing need for rationally designed 5-HT2AR agonists that possess optimal pharmacological profiles in order to fully reveal the therapeutic potential of these agonists and identify safer drug candidates devoid of hallucinogenic effects. This Perspective provides an overview of the structure-activity relationships of existing 5-HT2AR agonists based on their chemical classifications and discusses recent advancements in understanding their molecular pharmacology at a structural level. The encouraging clinical outcomes of psychedelics in depression treatment have sparked drug discovery endeavors aimed at developing novel 5-HT2AR agonists with improved subtype selectivity and signaling bias properties, which could serve as safer and potentially nonhallucinogenic antidepressants. These efforts can be significantly expedited through the utilization of structure-based methods and functional selectivity-directed screening.


Assuntos
Alucinógenos , Alucinógenos/farmacologia , Serotonina , Receptor 5-HT2A de Serotonina , Antidepressivos/farmacologia , Antidepressivos/uso terapêutico
2.
Mol Cell ; 66(5): 648-657.e4, 2017 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-28575660

RESUMO

The glycogen synthase kinase-3 (GSK3) family kinases are central cellular regulators highly conserved in all eukaryotes. In Arabidopsis, the GSK3-like kinase BIN2 phosphorylates a range of proteins to control broad developmental processes, and BIN2 is degraded through unknown mechanism upon receptor kinase-mediated brassinosteroid (BR) signaling. Here we identify KIB1 as an F-box E3 ubiquitin ligase that promotes the degradation of BIN2 while blocking its substrate access. Loss-of-function mutations of KIB1 and its homologs abolished BR-induced BIN2 degradation and caused severe BR-insensitive phenotypes. KIB1 directly interacted with BIN2 in a BR-dependent manner and promoted BIN2 ubiquitination in vitro. Expression of an F-box-truncated KIB1 caused BIN2 accumulation but dephosphorylation of its substrate BZR1 and activation of BR responses because KIB1 blocked BIN2 binding to BZR1. Our study demonstrates that KIB1 plays an essential role in BR signaling by inhibiting BIN2 through dual mechanisms of blocking substrate access and promoting degradation.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/efeitos dos fármacos , Brassinosteroides/farmacologia , Proteínas F-Box/metabolismo , Quinase 3 da Glicogênio Sintase/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Plantas Geneticamente Modificadas/efeitos dos fármacos , Proteínas Quinases/metabolismo , Esteroides Heterocíclicos/farmacologia , Ubiquitina-Proteína Ligases/metabolismo , Arabidopsis/enzimologia , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Sítios de Ligação , Domínio Catalítico , Proteínas de Ligação a DNA , Ativação Enzimática , Estabilidade Enzimática , Proteínas F-Box/genética , Genótipo , Quinase 3 da Glicogênio Sintase/genética , Mutação , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Fenótipo , Plantas Geneticamente Modificadas/enzimologia , Plantas Geneticamente Modificadas/genética , Complexo de Endopeptidases do Proteassoma/metabolismo , Ligação Proteica , Proteínas Quinases/genética , Proteólise , Transdução de Sinais/efeitos dos fármacos , Especificidade por Substrato , Ubiquitina-Proteína Ligases/genética , Ubiquitinação
3.
BMC Plant Biol ; 24(1): 44, 2024 Jan 11.
Artigo em Inglês | MEDLINE | ID: mdl-38200455

RESUMO

BACKGROUND: Hemerocallis citrina Baroni (Huang hua cai in Chinese) is a perennial herbaceous plant grown for its flower buds that are eaten fresh or dried and is known as the vegetarian three treasures. The nuclear genome of H. citrina has been reported, but the intraspecific variation of the plastome (plastid genome) has not yet been studied. Therefore, the panplastome of this species collected from diverse locations is reported here for the first time. RESULTS: In this study, 65 H. citrina samples were resequenced, de novo assembled, and aligned with the published plastome of H. citrina to resolve the H. citrina panplastome. The sizes of the 65 newly assembled complete plastomes of H. citrina ranged from 156,048 bp to 156,263 bp, and the total GC content ranged from 37.31 to 37.34%. The structure of the complete plastomes showed a typical tetrameric structure, including a large single copy (LSC), a small single copy (SSC), and a pair of inverted repeat regions (IRA and IRB). Many nucleotide variants were identified between plastomes, among which the variants in the intergenic spacer region were the most abundant, with the highest number of variants concentrated in the LSC region. Based on the phylogenetic tree constructed using the ML method, population structure analysis, and principal component analysis (PCA), the panplastome data were subdivided into five genetic clusters. The C5 genetic cluster was mostly represented by samples from Qidong, Hunan Province, while samples from Shanxi and Shaanxi Provinces were classified into the C4 genetic cluster. The greatest genetic diversity was found in the C1 genetic cluster, and the greatest genetic distance between any two clusters was found between the C4 and C5 clusters. CONCLUSION: The resolution of the panplastome and the analysis of the population structure of H. citrina plastomes provide important data for future breeding projects and germplasm preservation.


Assuntos
Hemerocallis , Filogenia , Melhoramento Vegetal , DNA Intergênico , Variação Genética , Plantas Comestíveis
4.
Curr Psychol ; : 1-13, 2023 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-36776148

RESUMO

The COVID-19 pandemic has greatly affected economies around the world, causing record unemployment rates that have exacerbated the already prevalent job insecurity, thereby leading to psychological distress among many individuals. The present study aims to reveal the underlying mechanisms of psychological distress induced by the COVID-19 pandemic among job-insecure employees and to identify one of the emotion regulation strategies (i.e., cognitive reappraisal) as a protective factor that mitigates psychological distress. Drawing upon transactional theory, we proposed and tested a moderated mediation model, wherein cognitive reappraisal serves as a moderator of the direct and indirect relationship between job insecurity and psychological distress via stress appraisals (i.e., threat appraisal and challenge appraisal) based on data from 922 employees. Results show that threat appraisal and challenge appraisal positively and negatively mediate the relationship between job insecurity and psychological distress, respectively. Furthermore, as an emotion regulation strategy, cognitive reappraisal mitigates psychological distress among job-insecure employees, weakens the positive indirect effect of threat appraisal, but intensifies the negative indirect effect of challenge appraisal. Our findings suggest that cognition plays an important role in individuals' emotional reactions to COVID-19-related stress, and cognition reappraisal is an effective emotion regulation strategy in mitigating psychological distress. Therefore, cognition reappraisal skills should be improved to reduce psychological distress induced by COVID-19 pandemic.

5.
Anal Biochem ; 659: 114950, 2022 12 15.
Artigo em Inglês | MEDLINE | ID: mdl-36243134

RESUMO

Herein, a signal amplified electrochemical immunosensor for the sensitive detection of cytokeratin 19 fragments (CYFRA 21-1) in human serum was discussed. The CoNi-RGO was used as a substrate for the sensor with excellent specific surface area and strong electrical conductivity, which enables more efficient attachment of antibodies. The introduction of the bimetallic sulfide NiCo2S4 composite ZIF material provides strong catalytic performance for the immunosensor. It is worth noting that, in addition to these satisfactory advantages, these two materials also show amazing signal amplification capacity. When the immunosensor works, the increase in electrical impedance decreases the electron transfer rate, making the electrochemical signal change obvious. The signal enhancement of immunosensors was emphasized by the marker during construction, and the experimental results were satisfactory. The proposed signal enhanced immunosensor had a linear relationship in the range of 0.001-10 ng/mL for CYFRA 21-1, and the minimum detection limit was 0.33 pg/mL for △I = 95.22 + 23.27 lg c. This demonstrates that the electrochemical immunosensor we constructed is successful and has a great developing potential.


Assuntos
Técnicas Biossensoriais , Caramujo Conus , Grafite , Nanopartículas Metálicas , Animais , Humanos , Técnicas Biossensoriais/métodos , Cobalto/química , Técnicas Eletroquímicas/métodos , Ouro , Imunoensaio/métodos , Queratina-19 , Limite de Detecção
6.
Mikrochim Acta ; 189(3): 126, 2022 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-35230535

RESUMO

In order to determine procalcitonin, a sandwich-type ratiometic electrochemical immunosensor was developed by differential pulse voltammetry (DPV). Due to high chemical stability and good biocompatibility, graphitic carbon nitride (g-C3N4) could be used as feasible supporter to carry silver nanoparticles (Ag NPs) with an obvious oxidative peak (measured typically at + 0.3 V vs. SCE). Ag NPs loaded onto g-C3N4 were not only beneficial to prevent the agglomeration of Ag NPs, but also favorable to improve the electron transfer velocity of g-C3N4. Moreover, the g-C3N4-Ag NPs as the matrix could immobilize primary antibody by Ag-N bond. Nile blue A (NBA), an excellent redox probe based on the redox reaction with two-electrons, provides a current signal at - 0.38 V (vs. SCE). Zr-based metal organic framework (UiO-67), an ideal framework material with large specific surface area and high porosity, could absorb the substantial water-soluble NBA by electrostatic adsorption. The UiO-67 modified by NBA (NBA-UiO-67) owned admirable biocompatibility and was a qualifying marker to load the secondary antibody. For the immunosensor, the current ratio of NBA to Ag NPs (INBA/IAg NPs) was increased as the concentrations of PCT increased. Under the optimum conditions, the linear range of the immunosensor was 0.005 to 50 ng/mL; the detection limit was 1.67 pg/mL (S/N = 3), which reflected the excellent analytical performance of the sensor. The proposed immunosensor strategy is a simple and dependable platform, with great application potential in biometric analysis.


Assuntos
Técnicas Biossensoriais , Nanopartículas Metálicas , Técnicas Eletroquímicas , Imunoensaio , Limite de Detecção , Nanopartículas Metálicas/química , Oxazinas , Pró-Calcitonina , Prata
7.
Small ; 17(25): e2100437, 2021 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-33991064

RESUMO

Self-assembled block copolymers are promising templates for fabricating thin film materials with tuned periodic feature sizes and geometry at the nanoscale. Here, a series of nanostructured platinum and iridium oxide electrocatalysts templated from poly(styrene)-block-poly(vinyl pyridine) (PSbPVP) block copolymers via an incipient wetness impregnation (IWI) pathway is reported. Both nanowire and nanocylinder electrocatalysts of varying feature sizes are assessed and higher catalyst loadings are achieved by the alkylation of the pyridine moieties in the PVP block prior to IWI. Electrocatalyst evaluations featuring hydrogen pump and water electrolysis demonstrations are carried out on interdigitated electrode (IDE) chips flexible with liquid supporting electrolytes and thin film polymer electrolytes. Notably, the mass activities of the nanostructured electrocatalysts from alkylated block copolymer templates are 35%-94% higher than electrocatalysts from non-alkylated block copolymer templates. Standing cylinder nanostructures lead to higher mass activities than lamellar variants despite their not having the largest surface area per unit catalyst loading demonstrating that mesostructure architectures have a profound impact on reactivity. Overall, IDE chips with model thin film electrocatalysts prepared from self-assembled block copolymers offer a high-throughput experimental method for correlating electrocatalyst nanostructure and composition to electrochemical reactivity.

8.
J Med Virol ; 92(9): 1556-1561, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32222119

RESUMO

The aim is to evaluate pregnant women infected with coronavirus disease 2019 (COVID-19) and provide help for clinical prevention and treatment. All five cases of pregnant women confirmed COVID-19 were collected among patients who admitted to the Maternal and Child Hospital of Hubei Province between January 20 and February 10, 2020. All patients, aging from 25 to 31 years old, had the gestational week from 38th weeks to 41st weeks. All pregnant women did not have an antepartum fever but developed a low-grade fever (37.5℃-38.5℃) within 24 hours after delivery. All patients had normal liver and renal function, two patients had elevated plasma levels of the myocardial enzyme. Unusual chest imaging manifestations, featured with ground-grass opacity, were frequently observed in bilateral (three cases) or unilateral lobe (two cases) by computed tomography (CT) scan. All labors smoothly processed, the Apgar scores were 10 points 1 and 5 minutes after delivery, no complications were observed in the newborn. Pregnancy and perinatal outcomes of patients with COVID-19 should receive more attention. It is probable that pregnant women diagnosed with COVID-19 have no fever before delivery. Their primary initial manifestations were merely low-grade postpartum fever or mild respiratory symptoms. Therefore, the protective measures are necessary on admission; the instant CT scan and real-time reverse-transcriptase polymerase-chain-reaction assay should be helpful in early diagnosis and avoid cross-infection on the occasion that patients have fever and other respiratory signs.


Assuntos
COVID-19/diagnóstico , COVID-19/virologia , Complicações Infecciosas na Gravidez/diagnóstico , Complicações Infecciosas na Gravidez/virologia , SARS-CoV-2 , Adulto , COVID-19/terapia , Gerenciamento Clínico , Feminino , Humanos , Avaliação de Resultados da Assistência ao Paciente , Reação em Cadeia da Polimerase , Gravidez , Complicações Infecciosas na Gravidez/terapia , Resultado da Gravidez , RNA Viral , SARS-CoV-2/genética , Avaliação de Sintomas , Tomografia Computadorizada por Raios X
9.
BMC Pregnancy Childbirth ; 20(1): 580, 2020 Oct 02.
Artigo em Inglês | MEDLINE | ID: mdl-33008308

RESUMO

BACKGROUND: During the ongoing global outbreak of COVID-19, pregnant women who are susceptible to COVID-19 should be highly concerned. The issue of vertical transmission and the possibility of neonatal infection is a major concern. CASE PRESENTATION: Case 1: A 35-year-old pregnant woman with a gestational age of 37 weeks and 6 days was admitted to our hospital at the point of giving birth. Except for the abnormalities in her chest CT image, she was asymptomatic. She had an uncomplicated spontaneous vaginal delivery, and her infant was discharged home for isolation. Because of the positive result of the maternal swabs for SARS-CoV-2 obtained on the 2nd day after sampling, we transferred the mother to the designated hospital and followed up with her by telephone interviews. Luckily, it was confirmed on February 23 that the newborn did not develop any COVID-19 symptoms after observation for 14 days after birth. Case 2: Another pregnant woman, with a gestational age of 38 weeks and 2 days, was also admitted to our hospital because of spontaneous labor with cervical dilation of 5 cm. Since she had the typical manifestations of COVID-19, including cough, lymphopenia, and abnormal chest CT images, she was highly suspected of having COVID-19. Based on the experience from case 1, we helped the mother deliver a healthy baby by vaginal delivery. On the 2nd day after delivery, the maternal nasopharyngeal swab result was positive, while the infant's result was negative. CONCLUSION: There is still insufficient evidence supporting maternal-fetal vertical transmission for COVID-19-infected mothers in late pregnancy, and vaginal delivery may not increase the possibility of neonatal infection.


Assuntos
Infecções Assintomáticas , Infecções por Coronavirus/diagnóstico , Parto Obstétrico/métodos , Pulmão/diagnóstico por imagem , Pneumonia Viral/diagnóstico , Complicações Infecciosas na Gravidez/diagnóstico , Adulto , Antibacterianos/uso terapêutico , Antivirais/uso terapêutico , Betacoronavirus , Aleitamento Materno , COVID-19 , Teste para COVID-19 , Técnicas de Laboratório Clínico , Infecções por Coronavirus/terapia , Tosse , Medicamentos de Ervas Chinesas/uso terapêutico , Feminino , Idade Gestacional , Humanos , Linfopenia , Máscaras , Oxigenoterapia , Pandemias , Isolamento de Pacientes , Equipamento de Proteção Individual , Pneumonia Viral/terapia , Reação em Cadeia da Polimerase , Gravidez , Complicações Infecciosas na Gravidez/terapia , SARS-CoV-2 , Testes Sorológicos , Tomografia Computadorizada por Raios X
10.
Ecotoxicol Environ Saf ; 171: 122-129, 2019 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-30597316

RESUMO

To explore the toxicity and action mechanism of acute sulfur dioxide (SO2) on urban landscape plants, a simulated SO2 stress environment by using fumigation chamber involving increasing SO2 concentration (0, 25, 50, 100, 200 mg m-3) was carried out among three species. After 72 h of exposure, SO2-induced oxidative damage indicated by electrolyte leakage increased with higher dose of SO2. Meanwhile, SO2 decreased the contents of chlorophyll a, chlorophyll b and carotenoid and increased the contents of sulfur. Net photosynthetic rate (Pn) decreased as a result of stomatal closure when SO2 dose was lower than 50 mg m-3, out of this range, non-stomatal limitation play a dominant role in the decline of Pn. Simultaneous measurements of chlorophyll fluorescence imaging (CFI) also revealed that the maximal quantum efficiency of PSII photochemistry in dark-adapted state (Fv/Fm) and the realized operating efficiency of PSII photochemistry (Fq'/Fm') was reduced by SO2 in a dose-dependent manner. In addition, the maximum quantum efficiency of PSII photochemistry in light-adapted state (Fv'/Fm') and the PSII efficiency factor (Fq'/Fv') decreased when exposure to SO2. These results implied that acute SO2 exposure induced photoinhibition of PSII reaction centers in landscape plants. Our study also indicated that different urban landscape plant species resist differently to SO2: Euonymus kiautschovicus > Ligustrum vicaryi > Syringa oblata according to gas-exchange characteristics and chlorophyll fluorescence responses.


Assuntos
Euonymus/efeitos dos fármacos , Ligustrum/efeitos dos fármacos , Dióxido de Enxofre/toxicidade , Syringa/efeitos dos fármacos , Dióxido de Carbono/metabolismo , Clorofila/metabolismo , Clorofila A/metabolismo , Euonymus/fisiologia , Fluorescência , Ligustrum/fisiologia , Fotossíntese/efeitos dos fármacos , Fotossíntese/fisiologia , Complexo de Proteína do Fotossistema II/metabolismo , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Syringa/fisiologia
11.
Ecotoxicol Environ Saf ; 111: 117-22, 2015 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-25450923

RESUMO

Sulfur dioxide (SO2) exposure is associated with increased risk of various damages to plants. However, little is known about the defense response in ornamental plants. In this study, an artificial fumigation protocol was carried out to study the defense potential of the glutathione (GSH)-ascorbate (AsA) dependent detoxification pathway to SO2 exposure in Tagetes erecta. The results show that when the plants were exposed to different doses of SO2 (0, 15, 30, 50 or 80 mg m(-3)) for different times (6, 12, 18, 24 or 33 h), SO2 induced oxidative stress was confirmed by the increased hydrogen peroxide (H2O2), malondialdehyde (MDA) and relative conductivity of membrane (RC) in a dose-dependent manner for different exposure times. However, the increased levels for H2O2, MDA and RC were not significant vis-a-vis the control when SO2 doses and exposure times were lower than 15 mg m(-3)/33 h, 30 mg m(-3)/24 h or 50 mg m(-3)/12 h (p>0.05). The results could be explained by the increases in the content of reduced form of glutathione (GSH), total glutathione (TGSH), ascorbate (AsA), ratio of GSH/GSSG (oxidized form of glutathione), activities of ascorbate peroxidase (APX), glutathione peroxidase (GPX), glutathione reductase (GR) and glutathione S-transferases (GST). On the other hand, exposure to higher doses of SO2 and longer exposure times, the values of the GSH-AsA dependent antioxidative indices decreased significantly (p<0.01), manifested by increased levels of H2O2. Furthermore, the levels of H2O2, MDA and RC varied little when SO2 doses and exposure times reached a 'critical' value (50 mg m(-3)/24 h). The defense ability of T. erecta to SO2 reached nearly extremity. To summarize, the response of T. erecta to elevated SO2 was related to higher H2O2 levels. GSH-AsA dependent detoxification pathway played an important role in against SO2-induced toxicity, although the defense response could not sufficiently alleviate oxidative damage when SO2 doses and exposure times reached critical value.


Assuntos
Poluentes Atmosféricos/toxicidade , Ácido Ascórbico/metabolismo , Glutationa/metabolismo , Dióxido de Enxofre/toxicidade , Tagetes/efeitos dos fármacos , Ascorbato Peroxidases/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Redutase/metabolismo , Peróxido de Hidrogênio/metabolismo , Malondialdeído/metabolismo , Oxirredução , Estresse Oxidativo , Tagetes/enzimologia , Tagetes/metabolismo
12.
Ecotoxicol Environ Saf ; 114: 198-203, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25645141

RESUMO

Both nitric oxide (NO) and reactive oxygen species (ROS) are very important signal molecules, but the roles they play in signal transduction of sulfur dioxide (SO2) induced toxicities on ornamental plants is not clear. In this study, the functions of NO and ROS in SO2-induced death of lower epidermal guard cells in ornamental plant Tagetes erecta were investigated. The results showed that SO2 derivatives (0.4-4.0 mmol L(-1) of final concentrations) could reduce the guard cells' viability and increase their death rates in a dose-dependent manner. Meanwhile, the significant increase of cellular NO, ROS, and Ca(2+) levels (P<0.05) and typical apoptosis features including nucleus condensation, nucleus break and nucleus fragmentation were observed. However, exposure to 2.0 mmol L(-1) of SO2 derivatives combined with either NO antagonists (NO scavenger c-PTIO; nitrate reductase inhibitor NaN3; NO synthase inhibitor L-NAME), ROS scavenger (AsA or CAT) or Ca(2+) antagonists (Ca(2+) scavenger EGTA or plasma membrane Ca(2+) channel blocker LaCl3) can effectively block SO2-induced guard cells death and corresponding increase of NO, ROS and Ca(2+) levels. In addition, addition of L-NAME or AsA in 2.0 mmol L(-1) of SO2 derivatives led to significant decrease in the levels of NO, ROS and Ca(2+), whereas addition of LaCl3 in them just resulted in the decrease of Ca(2+) levels, hardly making effects on NO and ROS levels. It was concluded that NO and ROS were involved in the apoptosis induced by SO2 in T. erecta, which regulated the cell apoptosis at the upstream of Ca(2+).


Assuntos
Poluentes Atmosféricos/toxicidade , Apoptose/efeitos dos fármacos , Óxido Nítrico/metabolismo , Epiderme Vegetal/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Dióxido de Enxofre/toxicidade , Tagetes/efeitos dos fármacos , Cálcio/metabolismo , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/farmacologia , Epiderme Vegetal/citologia , Epiderme Vegetal/metabolismo , Transdução de Sinais/efeitos dos fármacos , Tagetes/citologia , Tagetes/metabolismo
13.
Prep Biochem Biotechnol ; 44(3): 257-65, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24274014

RESUMO

The exogenous fragment sequence and flanking sequence between the exogenous fragment and recombinant chromosome of transgenic wheat B102-1-2 were successfully acquired using genome walking technology. The newly acquired exogenous fragment encoded the full-length sequence of transformed genes with transformed plasmid and corresponding functional genes including ubi, vector pBANF-bar, vector pUbiGUSPlus, vector HSP, reporter vector pUbiGUSPlus, promoter ubiquitin, and coli DH1. A specific polymerase chain reaction (PCR) identification method for transgenic wheat B102-1-2 was established on the basis of designed primers according to flanking sequence. This established specific PCR strategy was validated by using transgenic wheat, transgenic corn, transgenic soybean, transgenic rice, and non-transgenic wheat. A specifically amplified target band was observed only in transgenic wheat B102-1-2. Therefore, this method is characterized by high specificity, high reproducibility, rapid identification, and excellent accuracy for the identification of transgenic wheat B102-1-2.


Assuntos
DNA de Plantas/genética , Plantas Geneticamente Modificadas/genética , Análise de Sequência de DNA/métodos , Triticum/genética , Sequência de Bases , Passeio de Cromossomo , Reação em Cadeia da Polimerase/métodos
14.
Wei Sheng Wu Xue Bao ; 54(7): 737-45, 2014 Jul 04.
Artigo em Zh | MEDLINE | ID: mdl-25252454

RESUMO

OBJECTIVE: The effect of flhDC, fliA, fliD and fliE genes involved in moving of Escherichia coli (E. coli) on the motility of lysogened strain by Stx2-encoding phage phiMin27 was explored by gene knockout and phage lysogenic conversion. METHODS: Using the lambda Red recombinase system, the mutant strains of E. coli MG1655 named MG1655 deltaflhDC, MG1655 deltafliA, MG1655 deltafliD and MG1655 deltafliE were constructed. Then the corresponding complemented strains by ligating amplified targeted genes into the low copy vector pUC18 at the BamHI and Hind III sites and transforming these plasmids into mutant strains were acquired. By lysogenic infection of Stx2-encoding phage phiMin27, the lysogens for mutants named MG1655 deltaflhDCphiMin27, MG1655 deltafliAdeltaMin27, MG1655 deltafliDphiMin27 and MG1655 deltafliEphiMin27 were achieved. Subsequently, the motility of wild strain, the mutants, the complemented strains and the lysogens were detected. The changes of expression of the other genes involved in motility between wild strain and the lysogens before and after flhDC deletion by qRT-PCR were analyzed. RESULTS: Lysogenic infection of Stx2-encoding phage phiMin27 could promote the expression of fliA and fliD gene and enhance the motility of MG1655. For flhDC deletion, higher expression of fliA and fliD gene of MG1655 appeared, but the motility had no change. However, lysogen for MG1655 deltaflhDC lost the swimming motility. By gene transcriptional level detection, the expression of fliA and fliD gene of MG1655 deltaflhDCphiMin27 was down-regulated significantly compared with MG1655 deltaflhDC, and no marked variation was observed for fliE gene. The single deletion of fliA, fliD and fliE gene had no effect on the motility of E. coli MG1655 and lysogened strain by Stx2-encoding phage phiMin27. CONCLUSION: The results show that fliA and fliD gene together participated the regulation for flagella motility and flhDC gene could affect the motility of the lysogened strain by phage. It provides the theoretical basis for further research on the mutual regulation between phage lysogenization and host genes.


Assuntos
Bacteriófagos/fisiologia , Toxina Shiga II/biossíntese , Escherichia coli Shiga Toxigênica/citologia , Escherichia coli Shiga Toxigênica/virologia , Bacteriófagos/genética , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Flagelos/genética , Flagelos/fisiologia , Regulação Bacteriana da Expressão Gênica , Lisogenia , Toxina Shiga II/genética , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/metabolismo
15.
Neuron ; 112(4): 526-538, 2024 Feb 21.
Artigo em Inglês | MEDLINE | ID: mdl-38290517

RESUMO

Over the years, the field of GPCR drug design has undergone a remarkable evolution, fueled by advancements in science and technology. This evolution has given rise to a diverse range of ideas and approaches in structure-based drug design, bolstering the versatility and strength of the GPCR drug design toolbox. This review encapsulates the iterative development process, navigating challenges and opportunities in structure-based drug design within GPCRs. With a focused emphasis on its impact on psychiatric disorders, the review accentuates recent advancements and delves into the potentials unlocked by emerging technologies. The review explores the intricate interplay between scientific progress and iterative refinement, offering profound insights into the potential pathways that lie ahead for GPCR drug design.


Assuntos
Transtornos Mentais , Receptores Acoplados a Proteínas G , Humanos , Receptores Acoplados a Proteínas G/metabolismo , Desenho de Fármacos , Transtornos Mentais/tratamento farmacológico
16.
Anal Chim Acta ; 1288: 342056, 2024 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-38220266

RESUMO

Bacterial infections, viral infections and autoimmune diseases pose a considerable threat to human health. Procalcitonin (PCT) has emerged as a biomarker for the detection of these diseases. To ensure accurate and reliable results, we propose a dual-mode approach that incorporates self-validation and self-correction mechanisms. Herein, we develop a dual-mode self-powered photoelectrochemical (PEC) and colorimetric sensor to determine PCT. The self-powered PEC sensor was constructed with a photoanode of spherical nanoflower-MoS2/Cu2ZnSnS4/Bi2S3 material and a photocathode of CuInS2 material. Ni4Cu2 bimetallic hollow nanospheres (BHNs) possess superoxide dismutase and catalase performance, which facilitate superoxide anion radical (·O2-) and H2O2 circulating generation, promoting the separation of photogenerated electrons and holes to amplify photocurrent signal. Thus Ni4Cu2 BHNs is used as a marker material for PEC sensor. Meanwhile, in colorimetric mode, Ni4Cu2 BHNs converts blue oxTMB to a colourless TMB for colorimetric detection of PCT. Based on this principle, dual-mode determination of PCT with high sensitivity is achieved. The dual-mode method not only demonstrates outstanding properties and practicability, but also presents an effective, highly efficient and reliable method for detecting PCT.


Assuntos
Técnicas Biossensoriais , Nanosferas , Humanos , Nanosferas/química , Pró-Calcitonina , Molibdênio/química , Peróxido de Hidrogênio , Colorimetria , Técnicas Eletroquímicas/métodos , Técnicas Biossensoriais/métodos , Limite de Detecção
17.
J Cell Mol Med ; 17(7): 863-72, 2013 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-23672602

RESUMO

Stomatin is an important lipid raft-associated protein which interacts with membrane proteins and plays a role in the membrane organization. However, it is unknown whether it is involved in the response to hypoxia and glucocorticoid (GC) in alveolar epithelial cells (AEC). In this study we found that hypoxia and dexamethasone (dex), a synthetic GC not only up-regulated the expression of stomatin alone, but also imposed additive effect on the expression of stomatin in A549 cells, primary AEC and lung of rats. Then we investigated whether hypoxia and dex transcriptionally up-regulated the expression of stomatin by reporter gene assay, and found that dex, but not hypoxia could increase the activity of a stomatin promoter-driven reporter gene. Further deletion and mutational studies demonstrated that a GC response element (GRE) within the promoter region mainly contributed to the induction of stomatin by dex. Moreover, we found that hypoxia exposure did not affect membrane-associated actin, but decreased actin in cytoplasm in A549 cells. Inhibiting stomatin expression by stomatin siRNA significantly decreased dense of peripheral actin ring in hypoxia or dex treated A549 cells. Taken all together, these data indicated that dex and/or hypoxia significantly up-regulated the expression of stomatin in vivo and in vitro, which could stabilize membrane-associated actin in AEC. We suppose that the up-regulation of stomatin by hypoxia and dex may enhance the barrier function of alveolar epithelia and mediate the adaptive role of GC to hypoxia.


Assuntos
Actinas/metabolismo , Membrana Celular/metabolismo , Dexametasona/farmacologia , Células Epiteliais/citologia , Proteínas de Membrana/metabolismo , Alvéolos Pulmonares/citologia , Animais , Hipóxia Celular , Linhagem Celular Tumoral , Células Epiteliais/metabolismo , Genes Reporter , Humanos , Neoplasias Pulmonares/metabolismo , Mutagênese Sítio-Dirigida , Regiões Promotoras Genéticas , Ratos , Ratos Sprague-Dawley , Regulação para Cima
18.
Acta Biochim Biophys Sin (Shanghai) ; 45(5): 416-21, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23449073

RESUMO

In order to establish a specific identification method for genetically modified (GM) wheat, exogenous insert DNA and flanking sequence between exogenous fragment and recombinant chromosome of GM wheat B73-6-1 were successfully acquired by means of conventional polymerase chain reaction (PCR) and thermal asymmetric interlaced (TAIL)-PCR strategies. Newly acquired exogenous fragment covered the full-length sequence of transformed genes such as transformed plasmid and corresponding functional genes including marker uidA, herbicide-resistant bar, ubiquitin promoter, and high-molecular-weight gluten subunit. The flanking sequence between insert DNA revealed high similarity with Triticum turgidum A gene (GenBank: AY494981.1). A specific PCR detection method for GM wheat B73-6-1 was established on the basis of primers designed according to the flanking sequence. This specific PCR method was validated by GM wheat, GM corn, GM soybean, GM rice, and non-GM wheat. The specifically amplified target band was observed only in GM wheat B73-6-1. This method is of high specificity, high reproducibility, rapid identification, and excellent accuracy for the identification of GM wheat B73-6-1.


Assuntos
DNA de Plantas/genética , Plantas Geneticamente Modificadas/genética , Triticum/genética , DNA de Plantas/análise , Reação em Cadeia da Polimerase , Análise de Sequência de DNA/métodos
19.
Ecotoxicol Environ Saf ; 98: 41-5, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24125868

RESUMO

Chronic and acute exposure to SO2 is associated with increased risks of various damages to plants. In the present study, epidermal strip experiment was employed to investigate SO2-induced guard cells apoptosis and the signal regulation in Hemerocallis fulva. The results showed that with the increase of treatment concentrate of SO2 derivates (a mixture of sodium sulfite and sodium bisulfite, 3:1, mmol L⁻¹/mmol L⁻¹, 1.0-5.0 mmol L⁻¹), the physiological activity of the guard cells declined and cell death occurred. While the concentration of SO2 derivatives exceeded 2.0 mmol L⁻¹, the percentage of cell death increased significantly (P<0.05). Typical features of apoptosis including nuclear condensation, nuclear elongation, fragmentation etc. were found. Meanwhile, concomitant presence of nitric oxide (NO), reactive oxygen species (ROS) and Ca²âº level increment appeared. However, SO2-induced cell death can be effectively blocked by either of the following substances with their respective optimal concentrations: antioxidant ascorbic acid (Asc; 0.05 mmol L⁻¹) or catalase (CAT; 200 U mL⁻¹), nitric oxide (NO) scavenger 2-(4-carboxyphenyl)-4, 4, 5, 5- tetramethylmidiazoline-1-oxyl-3-oxide (c-PTIO; 0.20 mmol L⁻¹), nitrate reductase inhibitor NaN3 (0.20 mmol L⁻¹), Ca²âº chelating agent EGTA (0.05 mmol L⁻¹) or plasma membrane Ca²âº channel blocker LaCl3 (0.05 mmol L⁻¹). In addition to a significant decrease in cell death rate, a reduction in the levels of reactive oxygen species (ROS), NO and Ca²âº was observed. Further study showed that compared to treatment with SO2 alone, Asc treatment led to a decrease in NO and Ca²âº levels and NaN3 treatment led to a decrease in ROS and Ca²âº levels, but the NO and ROS levels of the LaCl3 treatment changed little. All results suggested that NO, ROS and Ca²âº were involved in the apoptosis induced by SO2 in H. fulva. The process might be related to the burst of NO or ROS, which would activate the plasma Ca²âº channel and result in the increase of intercellular Ca²âº.


Assuntos
Apoptose/efeitos dos fármacos , Hemerocallis/efeitos dos fármacos , Dióxido de Enxofre/toxicidade , Cálcio/metabolismo , Núcleo Celular/ultraestrutura , Hemerocallis/citologia , Hemerocallis/metabolismo , Óxido Nítrico/metabolismo , Epiderme Vegetal/citologia , Epiderme Vegetal/efeitos dos fármacos , Epiderme Vegetal/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transdução de Sinais
20.
Talanta ; 260: 124632, 2023 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-37149935

RESUMO

In this study, a ratiometric electrochemical immunosensor has been developed to detect the cytokeratin 19 fragment 21-1 (CYFRA21-1) biomarker in a highly sensitive manner through a dual-signal output model. As one of signal indicators, snowflake-like FeSe2 loaded with AuNPs (FeSe2-AuNPs) as sensing substrate with good conductivity and large active sites provides a differential pulse voltammetry (DPV) signal at +0.4 V. Another signal indicator, toluidine blue (TB) with the water-solubility property is an excellent redox probe that can generate DPV signal at -0.3 V. To solve the water-solubility problem, the TB is absorbed with polyacrylic acid (PAA) functionalized ZIF-67 (PAA-ZIF-67), which retains the properties of ZIF-67 that are large specific surface area and strong adsorption properties. The ratio of signals, stemmed from PAA-ZIF@TB and FeSe2-AuNPs (IPAA-ZIF@TB/IFeSe2-AuNPs), increases with the CYFRA21-1 concentration. Under optimal experimental conditions, CYFRA21-1 was detected in a wide dynamic range from 0.1 pg/mL to 100 ng/mL, with a lower limit of detection of 0.02 pg/mL. Looking ahead, this ratio based strategy provides prospective clinical applications for detecting other biomarkers.


Assuntos
Técnicas Biossensoriais , Grafite , Nanopartículas Metálicas , Grafite/química , Técnicas Eletroquímicas , Ouro/química , Cloreto de Tolônio , Estudos Prospectivos , Imunoensaio , Nanopartículas Metálicas/química , Água , Limite de Detecção
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