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Natural variations in cis-regulatory regions often affect crop phenotypes by altering gene expression. However, the mechanism of how promoter mutations affect gene expression and crop stress tolerance is still poorly understood. In this study, by analyzing RNA-sequencing (RNA-Seq) data and reverse transcription quantitative real-time PCR validation in the cultivated tomato and its wild relatives, we reveal that the transcripts of WRKY33 are almost unchanged in cold-sensitive cultivated tomato Solanum lycopersicum L. 'Ailsa Craig' but are significantly induced in cold-tolerant wild tomato relatives Solanum habrochaites LA1777 and Solanum pennellii LA0716 under cold stress. Overexpression of SlWRKY33 or ShWRKY33 positively regulates cold tolerance in tomato. Variant of the critical W-box in SlWRKY33 promoter results in the loss of self-transcription function of SlWRKY33 under cold stress. Analysis integrating RNA-Seq and chromatin immunoprecipitation sequencing data reveals that SlWRKY33 directly targets and induces multiple kinases, transcription factors, and molecular chaperone genes, such as CDPK11, MYBS3, and BAG6, thus enhancing cold tolerance. In addition, heat- and Botrytis-induced WRKY33s expression in both wild and cultivated tomatoes are independent of the critical W-box variation. Our findings suggest nucleotide polymorphism in cis-regulatory regions is crucial for different cold sensitivity between cultivated and wild tomato plants.
Assuntos
Solanum lycopersicum , Solanum , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Chaperonas Moleculares/metabolismo , RNA/metabolismo , Solanum/genética , Solanum/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Polimorfismo de Nucleotídeo Único , Regiões Promotoras GenéticasRESUMO
The development of viable pollen determines male fertility, and is crucial for reproduction in flowering plants. Phytochrome interacting factor 3 (PIF3) acts as a central regulator of plant growth and development, but its relationship with pollen development has not been determined. Through genetic, histological and transcriptomic analyses, we identified an essential role for SlPIF3 in regulating tomato (Solanum lycopersicum) pollen development. Knocking out SlPIF3 using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 resulted in pollen mitosis I arrest, and a failure to form viable pollen. We further demonstrated that both glutamate synthase 1 (SlGLT1) and cell wall invertase 9 (SlCWIN9), involved in auxin and sugar homeostasis, respectively, colocalised with SlPIF3 in the anthers and were directly regulated by SlPIF3. Knockout of either SlGLT1 or SlCWIN9 phenocopied the pollen phenotype of SlPIF3 knockout (Slpif3) lines. Slpif3 fertility was partially restored by exogenous auxin indole-3-acetic acid in a dose-dependent manner. This study reveals a mechanism by which SlPIF3 regulates pollen development and highlights a new strategy for creating hormone-regulated genic male sterile lines for tomato hybrid seed production.
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Fitocromo , Solanum lycopersicum , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos/metabolismo , Solanum lycopersicum/metabolismo , Fitocromo/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Pólen/metabolismo , Açúcares/metabolismoRESUMO
Drought limits the growth and productivity of plants. Reproductive development is sensitive to drought but the underlying physiological and molecular mechanisms remain unclear in tomatoes. Here, we investigated the effect of drought on tomato floral development using morpho-physiological and transcriptome analyses. Drought-induced male sterility through abnormal anther development includes pollen abortion, inadequate pollen starch accumulation and anther indehiscence which caused floral bud and opened flower abortions and reduced fruit set/yield. Under drought stress (DS), pollen mother cell to meiotic (PMC-MEI) anthers survived whereas tetrad to vacuolated uninucleate microspore (TED-VUM) anthers aborted. PMC-MEI anthers had lower ABA increase, reduced IAA and elevated sugar contents under DS relative to well-watered tomato plants. However, TED-VUM anthers had higher ABA increase and IAA levels, and lower accumulation of soluble sugars, indicating abnormal carbohydrate and hormone metabolisms when exposed to drought-stress conditions. Moreover, RNA-Seq analysis identified altogether >15,000 differentially expressed genes that were assigned to multiple pathways, suggesting that tomato anthers utilize complicated mechanisms to cope with drought. In particular, we found that tapetum development and ABA homeostasis genes were drought-induced while sugar utilization and IAA metabolic genes were drought-repressed in PMC-MEI anthers. Our results suggest an important role of phytohormones metabolisms in anther development under DS and provide novel insight into the molecular mechanism underlying drought resistance in tomatoes.
Assuntos
Secas , Flores/anatomia & histologia , Flores/fisiologia , Solanum lycopersicum/crescimento & desenvolvimento , Solanum lycopersicum/genética , Estresse Fisiológico/genética , Transcriptoma/genética , Transporte Biológico , Fertilidade , Flores/citologia , Flores/genética , Frutas/genética , Frutas/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Modelos Biológicos , Reguladores de Crescimento de Plantas/metabolismo , Pólen/citologia , Pólen/genética , Pólen/ultraestrutura , Transdução de Sinais , Solubilidade , Amido/metabolismo , Sacarose/metabolismoRESUMO
Substantial evidence indicates that microRNAs (miRNAs) can be used as biological markers of Parkinson's disease (PD) and contribute to the risk assessment, early diagnosis, and treatment. We aimed to explore the role and potential mechanism of miR-20a-5p on inflammation and oxidative stress in 1-methyl-4-phenyl pyridine ion- (MPP+-) induced HT22 cells. HT22 cells were pretreated with miR-20a-5p mimic and/or pcDNA-IRF9 for 24 h and then treated with MPP+ (0.5 mM) for 24 h. The cell viability and apoptosis were determined using the Cell Counting Kit-8 (CCK-8) and Annexin V FITC/PI staining flow cytometry assay, respectively. The expression and secretion of inflammatory factors and oxidative stress-related factors were detected by enzyme-linked immunosorbent assay (ELISA). The protein expression levels were detected using Western blot analysis. Here, we discovered that MPP+ led to mitochondrial dysfunction, inflammation, and cell damage of HT22 cells, which were alleviated by miR-20a-5p overexpression. We further clarified that interferon regulatory factor 9 (IRF9) was a target gene of miR-20a-5p. IRF9 contributed to MPP+-induced mitochondrial disruption, inflammation, and cell apoptosis. Moreover, IRF9 hindered the improvement of miR-20a-5p overexpression on MPP+-induced neurotoxicity. Furthermore, the decrease of p-P65 level induced by miR-20a-5p mimic was significantly reversed by IRF9 overexpression. These findings demonstrate that miR-20a-5p contributes to MPP+-induced mitochondrial disruption and cell damage, and miR-20a-5p might be a novel therapeutic target for PD.
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INTRODUCTION: Coronavirus disease 2019 (COVID-19) is a rapidly emerging infectious respiratory disease caused by severe acute respiratory syndrome coronavirus 2. Currently, more than 100 million cases of COVID-19 have been confirmed worldwide, with over 2.4 million mortalities. The pandemic affects people of all ages but older individuals and those with severe chronic illnesses, including cancer patients, are at higher risk. PATIENT CONCERNS: The impact of cancer treatment on the progression of COVID-19 is unclear. Therefore, we assessed the effects of chemotherapy on COVID-19 outcomes for 2 cancer patients. On January 24, 2020, a level I response to a major public health emergency was initiated in Hubei Province, China, which includes Enshi Autonomous Prefecture that has a population of 4.026 million people. As of April 30, 2020, 252 confirmed cases of COVID-19 and 11 asymptomatic carriers were identified in Enshi. DIAGNOSIS: Among the confirmed cases and asymptomatic carriers, 2 patients were identified who were previously diagnosed with malignant tumors, including one with hepatocellular carcinoma and the other with cardia carcinoma. INTERVENTIONS: These 2 patients were receiving or just completed chemotherapy at the time of their COVID-19 diagnosis. OUTCOMES: Both patients were followed and presented favorable outcomes. The positive outcomes for these 2 patients could be partially explained by their recent chemotherapy that impacted their immune status. Also, their relatively younger ages and lack of comorbidities were likely factors in their successful recovery from COVID-19. CONCLUSIONS: Anticancer treatment might enhance a patient's ability to respond favorably to COVID-19 infection. However, anticancer treatment is likely to impact immune function differently in different individuals, which can influence disease outcomes.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , COVID-19/imunologia , Neoplasias Hepáticas/tratamento farmacológico , SARS-CoV-2/imunologia , Neoplasias Gástricas/tratamento farmacológico , Adulto , COVID-19/complicações , COVID-19/diagnóstico , Teste de Ácido Nucleico para COVID-19 , Ciclobutanos/uso terapêutico , Docetaxel/uso terapêutico , Quimioterapia Combinada/métodos , Humanos , Neoplasias Hepáticas/complicações , Neoplasias Hepáticas/imunologia , Pulmão/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Compostos Organoplatínicos/uso terapêutico , RNA Viral/isolamento & purificação , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Sorafenibe/uso terapêutico , Neoplasias Gástricas/complicações , Neoplasias Gástricas/imunologia , Tomografia Computadorizada por Raios X , Resultado do Tratamento , Tratamento Farmacológico da COVID-19RESUMO
Nowadays, medicines from plant sources play a vital role in healthcare management. Chrysin, a plant flavonoid, possesses a wide range of pharmacological activities. The aim of present investigation was to evaluate the safety of chrysin by determining toxicity after acute and sub-chronic oral administration in rats. Acute oral toxicity (AOT) and sub-chronic oral toxicity studies of chrysin were carried out according to OECD 425 and OCED 408 in Sprague Dawley rats. In AOT, oral administration of chrysin (5000 mg/kg) showed 40% mortality. In the sub-chronic toxicity study, daily oral administration of chrysin (1000 mg/kg) showed significantly decreased body weight whereas liver weight was increased significantly in male rats. A significant alteration in the hematology (RBC, MCH, MCHC, TLC, lymphocytes, and neutrophil) and blood chemistry (albumin, bilirubin, ALT, AST, creatinine, and GGT) were found in chrysin (1000 mg/kg) treated rats which were either limited to one sex or lacked dose-response or were within the normal laboratory ranges. There was a significant increase in hepatic and renal oxido-nitrosative stress in chrysin (1000 mg/kg) treated rats. There was no significant change in electrocardiographic (except heart rate), hemodynamic, the left ventricular function, and lung function test. Renal and hepatic histological aberrations were induced in chrysin (1000 mg/kg) treated rats. In conclusion results of the present investigation determined the LD50 value of chrysin to be 4350 mg/kg whereas NOAEL and LOAEL of chrysin was found to be 500 and 1000 mg/kg, respectively for both the sexes.
Assuntos
Flavonoides , Extratos Vegetais , Administração Oral , Animais , Flavonoides/toxicidade , Dose Letal Mediana , Masculino , Ratos , Ratos Sprague-Dawley , Testes de Toxicidade AgudaRESUMO
Dietary factors play a crucial role in the management of type 2 diabetes mellitus (T2DM) by reducing cardiovascular disease (CVD) risk. Therefore, we aimed to examine the associations between habitual green tea consumption and risk factors of CVD among T2DM patients. A total of 1013 patients with T2DM were included in a community-based cross-sectional study. Data on dietary habits, including tea consumption, were collected using a food frequency questionnaire. A multivariable logistic regression model was used to analyze the associations. In men, as compared with nongreen tea drinkers, odds ratios (ORs) (95% confidence interval [CI]) of nonalcoholic fatty liver disease (NAFLD) were 2.06 (95% CI, 1.20-3.55) for those with green tea consumption of once per day and 2.45 (95% CI, 1.31-4.58) for more than or equal to twice per day (P-trend = .004); ORs (95% CI) of general obesity were 2.19 (95% CI, 1.02-4.68) and 2.70 (95% CI, 1.18-6.21), respectively (P-trend = .021); whereas no such association was found in women. Sensitivity analysis according to self-awareness of their T2DM status revealed that the positive association between green tea consumption and general obesity was not reliable. Higher intake of green tea was still positively associated with NAFLD, but it only persisted in participants aged ≥52 years or the lower dietary quality subgroup in further analyses. Our findings suggest that tea consumption was associated with an increased risk of NAFLD among male T2DM patients aged 52 years or older, and those with lower dietary quality, which needs to be confirmed in future prospective studies.
Assuntos
Doenças Cardiovasculares , Diabetes Mellitus Tipo 2 , Hepatopatia Gordurosa não Alcoólica , Chá/química , Doenças Cardiovasculares/epidemiologia , China/epidemiologia , Estudos Transversais , Diabetes Mellitus Tipo 2/epidemiologia , Feminino , Fatores de Risco de Doenças Cardíacas , Humanos , Masculino , Pessoa de Meia-Idade , Hepatopatia Gordurosa não Alcoólica/epidemiologia , Estudos ProspectivosRESUMO
Tumor-associated macrophages (TAMs) have been shown to be associated with poor prognosis of cancer and are predominately localized in the hypoxia regions of tumor. We demonstrated in this study that hypoxia increases the synthesis and secretion of galectin-3 by TAMs. The increased expression of galectin-3 in TAMs was seen to be associated with nucleation of transcription factor NF-κB through generation and activation of ROS and promoted tumor growth and metastasis in vitro and in mice through multiple molecular mechanisms. It was found that the TAMs-mediated promotion of tumor growth and metastasis in hypoxia was inhibited by administration of macrophage-depletion agent clodronate liposomal (CL) or galectin-3 inhibitor modified citric pectin (MCP) in orthotopic syngeneic mammary adenocarcinoma model and metastasis model. Co-administration of anti-angiogenesis agent sorafenib or bevacizumab with CL and MCP showed to cause stronger inhibition of tumor growth and metastasis than administration of each agent alone. These results indicate that hypoxia-induced galectin-3 expression and secretion from TAMs promotes tumor growth and metastasis. Targeting the actions of galectin-3 in hypoxia may be a potential therapeutic strategy for cancer treatment.
Assuntos
Adenocarcinoma/tratamento farmacológico , Bevacizumab/farmacologia , Neoplasias da Mama/tratamento farmacológico , Galectina 3/antagonistas & inibidores , Regulação Neoplásica da Expressão Gênica , Hipóxia/tratamento farmacológico , Neoplasias Mamárias Experimentais/tratamento farmacológico , Adenocarcinoma/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Animais , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Ácido Clodrônico/farmacologia , Técnicas de Cocultura , Progressão da Doença , Feminino , Galectina 3/genética , Galectina 3/metabolismo , Humanos , Hipóxia/genética , Hipóxia/metabolismo , Hipóxia/patologia , Metástase Linfática , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/patologia , Neoplasias Mamárias Experimentais/genética , Neoplasias Mamárias Experimentais/metabolismo , Neoplasias Mamárias Experimentais/patologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , NF-kappa B/genética , NF-kappa B/metabolismo , Neovascularização Patológica , Pectinas/farmacologia , Transdução de Sinais , Sorafenibe/farmacologiaRESUMO
Coronavirus Disease 2019 (COVID-19) caused by the emerging severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is associated with a crude case fatality rate of about 0.5-10 % depending on locality. A few clinically approved drugs, such as remdesivir, chloroquine, hydroxychloroquine, nafamostat, camostat, and ivermectin, exhibited anti-SARS-CoV-2 activity in vitro and/or in a small number of patients. However, their clinical use may be limited by anti-SARS-CoV-2 50 % maximal effective concentrations (EC50) that exceeded their achievable peak serum concentrations (Cmax), side effects, and/or availability. To find more immediately available COVID-19 antivirals, we established a two-tier drug screening system that combines SARS-CoV-2 enzyme-linked immunosorbent assay and cell viability assay, and applied it to screen a library consisting 1528 FDA-approved drugs. Cetilistat (anti-pancreatic lipase), diiodohydroxyquinoline (anti-parasitic), abiraterone acetate (synthetic androstane steroid), and bexarotene (antineoplastic retinoid) exhibited potent in vitro anti-SARS-CoV-2 activity (EC50 1.13-2.01 µM). Bexarotene demonstrated the highest Cmax:EC50 ratio (1.69) which was higher than those of chloroquine, hydroxychloroquine, and ivermectin. These results demonstrated the efficacy of the two-tier screening system and identified potential COVID-19 treatments which can achieve effective levels if given by inhalation or systemically depending on their pharmacokinetics.
Assuntos
Antivirais/farmacologia , Betacoronavirus , Infecções por Coronavirus/tratamento farmacológico , Avaliação Pré-Clínica de Medicamentos/métodos , Pneumonia Viral/tratamento farmacológico , Androstenos/farmacologia , Animais , Benzoxazinas/farmacologia , Betacoronavirus/efeitos dos fármacos , Betacoronavirus/fisiologia , Bexaroteno/farmacologia , COVID-19 , Células CACO-2 , Sobrevivência Celular/efeitos dos fármacos , Chlorocebus aethiops , Infecções por Coronavirus/virologia , Efeito Citopatogênico Viral/efeitos dos fármacos , Bases de Dados de Produtos Farmacêuticos , Aprovação de Drogas , Reposicionamento de Medicamentos , Ensaio de Imunoadsorção Enzimática , Humanos , Iodoquinol/farmacologia , Pandemias , Pneumonia Viral/virologia , SARS-CoV-2 , Estados Unidos , United States Food and Drug Administration , Células Vero , Carga Viral/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Tratamento Farmacológico da COVID-19RESUMO
OBJECTIVE: To evaluate the effect of transcutaneous electrical acupuncture point stimulation (TEAS) on sperm parameters and the underlying molecular mechanisms. METHODS: A total of 121 patients diagnosed with oligozoospermia, asthenozoospermia or oligoasthenozoospermia were randomised into four groups (three treatment groups, one control): the TEAS groups were treated with 2 Hz (n=31), 100 Hz (n=31), or mock stimulation (n=29) at acupuncture points BL23, ST36, CV1 and CV4 for 2 months. The control group (n=30) was provided with lifestyle advice only. RESULTS: The changes in total sperm count and motility in the 2 Hz TEAS group were significantly greater than those in the mock group and the control group. The change in neutral α-glucosidase (NAG) and zinc levels in the 2 Hz group were significantly greater than those in the mock group and control group, and the changes in fructose levels of the 2 Hz group were significantly greater than those in the control group. Significant increases in calcium and integrin-binding protein 1 (CIB1) and reduction of cyclin-dependent kinase 1 b (CDK1) were also found after 2 Hz TEAS treatment. CONCLUSIONS: The present findings suggest that 2 Hz TEAS can improve sperm count and motility in patients with abnormal semen parameters, and is associated with increases in seminal plasma zinc, NAG and fructose. The upregulation of CIB1 and downregulation of CDK1 by TEAS may be associated with its positive effects on sperm motility and count. TRIAL REGISTRATION: http://www.chictr.org ; registration no. ChiCTR-TRC-11001775.
Assuntos
Astenozoospermia/terapia , Eletroacupuntura , Oligospermia/terapia , Sêmen/metabolismo , Pontos de Acupuntura , Adulto , Astenozoospermia/metabolismo , Astenozoospermia/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Oligospermia/metabolismo , Oligospermia/fisiopatologia , Sêmen/citologia , Contagem de Espermatozoides , Motilidade dos Espermatozoides , Espermatozoides/citologia , Resultado do TratamentoRESUMO
High temperature (HT) is becoming an increasingly serious factor in limiting crop production with global climate change. During hot seasons, owing to prevailing HT, cultivated tomatoes are prone to exhibiting stigma exsertion, which hampers pollination and causes fruit set failure. However, the underlying regulatory mechanisms of the HT-induced stigma exsertion remain largely unknown. Here, we demonstrate that stigma exsertion induced by HT in cultivated tomato is caused by more seriously shortened stamens than pistils, which is different from the stigma exsertion observed in wild tomato species. Under the HT condition, the different responses of pectin, sugar, expansin, and cyclin cause cell wall remodelling and differentially localized cell division and selective cell enlargement, which further determine the lengths of stamens and pistils. In addition, auxin and jasmonate (JA) are implicated in regulating cell division and cell expansion in stamens and pistils, and exogenous JA instead of auxin treatment can effectively rescue tomato stigma exsertion through regulating the JA/COI1 signalling pathway. Our findings provide a better understanding of stigma exsertions under the HT condition in tomato and uncover a new function of JA in improving plant abiotic stress tolerance.
Assuntos
Ciclopentanos/metabolismo , Flores/fisiologia , Oxilipinas/metabolismo , Reguladores de Crescimento de Plantas/metabolismo , Transdução de Sinais , Solanum lycopersicum/metabolismo , Flores/crescimento & desenvolvimento , Frutose/metabolismo , Glucose/metabolismo , Temperatura Alta , Ácidos Indolacéticos/metabolismo , Solanum lycopersicum/crescimento & desenvolvimento , Solanum lycopersicum/fisiologia , Pectinas/metabolismo , Polinização , Reação em Cadeia da Polimerase em Tempo Real , Autofertilização , Sacarose/metabolismoRESUMO
Zika virus (ZIKV) is a human-pathogenic flavivirus that has recently emerged as a global public health threat. ZIKV infection may be associated with congenital malformations in infected fetuses and severe neurological and systemic complications in infected adults. There are currently limited treatment options for ZIKV infection. AR-12 (OSU-03012) is a celecoxib derivative cellular kinase inhibitor that has broad-spectrum antiviral activities. In this study, we investigated the antiviral activity and mechanism of AR-12 against ZIKV. We evaluated the in vitro anti-ZIKV activity of AR-12, using cell protection and virus yield reduction assays, in multiple clinically relevant cell lines, and the in vivo treatment effects of AR-12 in a lethal mouse model using type I interferon receptor-deficient A129 mice. AR-12 inhibited ZIKV strains belonging to both the African and Asian/American lineages in Huh-7 and/or neuronal cells. AR12's IC50 against ZIKV was consistently <2⯵M in these cells. ZIKV-infected A129 mice treated with intraperitoneally or orally administered AR-12 had significantly higher survival rate (50.0%-83.3% vs 0%, Pâ¯<â¯0.05), less body weight loss, and lower blood and tissue ZIKV RNA loads than untreated control A129 mice. These anti-ZIKV effects were likely the results of down-regulation of the PI3K/Akt pathway by AR-12. Clinical trials using the clinically available and broad-spectrum AR-12 as an empirical treatment should be considered especially for patients residing in or returning from areas endemic of ZIKV and other arboviral infections who present with an acute undifferentiated febrile illness.
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Antivirais/farmacologia , Inibidores de Proteínas Quinases/farmacologia , Pirazóis/farmacologia , Sulfonamidas/farmacologia , Infecção por Zika virus/prevenção & controle , Zika virus/efeitos dos fármacos , Administração Oral , Animais , Antivirais/uso terapêutico , Linhagem Celular , Modelos Animais de Doenças , Humanos , Injeções Intraperitoneais , Camundongos , Testes de Sensibilidade Microbiana , Inibidores de Proteínas Quinases/uso terapêutico , Pirazóis/uso terapêutico , Sulfonamidas/uso terapêutico , Análise de Sobrevida , Resultado do Tratamento , Carga Viral , Zika virus/crescimento & desenvolvimento , Infecção por Zika virus/patologia , Infecção por Zika virus/virologiaRESUMO
OBJECTIVE: To observe the effects of Xiusanzhen treatment on the ultrastructure of olfactory bulb and the expression of substantia nigra glial fibrillary acidic protein (GFAP) in mice with Parkinson's disease (PD) induced by lipopolysaccharide (LPS), and to provide methods and evidence for early prevention and treatment of PD. METHODS: Forty C57BL/6 male mice were randomly divided into a blank group, a model group, an electroacupuncture (EA) group and a medication group, 10 mice in each one. The mice in the model group, EA group and medication group were treated with 30-day nasal perfusion of LPS to establish PD model. From the first day of model establishment, the mice in the EA group were treated with electroacupuncture at bilateral "Yingxiang" (LI 20) and "Yintang" (GV 29) for 20 min, once a day; 5-day treatment was taken as one session, and 4 sessions were given with an interval of 2 days between sessions. The mice in the medication group were treated with intraperitoneal injection of L-DOPA, 10 mg/mL, once a day; 5-day treatment was taken as one session, and 4 sessions were given with an interval of 2 days between sessions. After treatment, the behavioristics changes were observed by using footprint analysis and swimming test score; the ultrastructure of olfactory bulb was observed by using transmission electron microscopy; the expression of GFAP in substantia nigra was measured by using western blot method. RESULTS: â After model establishment, the mice in the model group, the EA group and medication group showed significant symptoms of quiver and fear of chill, and the BMI was significantly lower than that in the blank group (all P<0.05). The results of behavioristics test indicated footprint length and swimming time in the model group, the EA group and medication group was significantly lower than those in the blank group (all P<0.01), indicating the successful establishment of PD model. â¡ After one-session treatment, the symptoms of quiver and fear of chill were not observed in the EA group. After 4-session treatment, the BMI in the EA group was significantly higher than that in the model group (P<0.05); the symptoms of quiver and fear of chill were not observed in the medication group, but the BMI was similar with the model group (P>0.05). ⢠After treatment, the footprint and swimming time in the model group were significantly lower than that in the blank group (both P<0.01); the footprint and swimming time in the EA group and medication group were significantly higher than those in the model group (all P<0.01).⣠After treatment, compared with the blank group, the organelles and ultrastructure of olfactory bulb in the model group were significantly improved; the ultrastructure of olfactory bulb in the EA group was improved compared with that in the model group. ⤠After treatment, the expression of substantia nigra GFAP in the model group was significantly higher than that in the blank group (P<0.01); the expression of substantia nigra GFAP in the EA group and medication group was significantly lower than that in the model group (both P<0.05). CONCLUSION: The early treatment of Xiusanzhen can improve behavioral disorders in LPS-induced early PD mice, and the mechanism may be related to the regulation of olfactory disorders and the expression of GFAP in substantia nigra.
Assuntos
Eletroacupuntura , Doença de Parkinson , Animais , Proteína Glial Fibrilar Ácida , Masculino , Camundongos Endogâmicos C57BL , Bulbo Olfatório , Ratos , Ratos Sprague-DawleyRESUMO
We previously disclosed the identification of cereblon modulator 3 (CC-885), with potent antitumor activity mediated through the degradation of GSPT1. We describe herein the structure-activity relationships for analogs of 3 with exploration of the structurally related dioxoisoindoline class. The observed activity of protein degradation could in part be rationalized through docking into the previously disclosed 3-CRBN-GSPT1 cocrystal ternary complex. For SAR that could not be rationalized through the cocrystal complex, we sought to predict SAR through a QSAR model developed in house. Through these analyses, selective protein degradation could be achieved between the two proteins of interest, GSPT1 and Aiolos.
Assuntos
Fator de Transcrição Ikaros/metabolismo , Fatores de Terminação de Peptídeos/metabolismo , Proteólise/efeitos dos fármacos , Relação Quantitativa Estrutura-Atividade , Ubiquitina-Proteína Ligases/metabolismo , Proteínas Adaptadoras de Transdução de Sinal , Antineoplásicos/química , Antineoplásicos/farmacologia , Técnicas de Química Sintética , Avaliação Pré-Clínica de Medicamentos/métodos , Humanos , Fator de Transcrição Ikaros/química , Fator de Transcrição Ikaros/genética , Simulação de Acoplamento Molecular , Mieloma Múltiplo , Peptídeo Hidrolases/química , Peptídeo Hidrolases/metabolismo , Fatores de Terminação de Peptídeos/química , Fatores de Terminação de Peptídeos/genética , Ftalimidas/química , Piperidonas/químicaRESUMO
Diabetic gastroparesis (DGP) is an important complication of diabetes, and seriously impacts the patient's quality of life. In the present paper, the authors summarize national and international research on the mechanism of acupuncture therapy in improving DGP and the pathogenesis of DGP over the past 10 years. The authors illustrate the mechanism of acupuncture underlying the improvement in DGP as follows:1) the effect on autonomic nerves and neurons of the central nervous system; 2) promoting restoration of the enteric nervous system; 3) the effect on Cajal interstitial cells; 4) improving gastrointestinal hormone disorder; 5) improving hyperglycemic state; 6) the effect on gastric smooth muscle. The authors also put forward some suggestions for future research on the mechanisms of acupuncture in DGP.
Assuntos
Terapia por Acupuntura , Diabetes Mellitus , Gastroparesia/terapia , Humanos , Qualidade de VidaRESUMO
Ion channels regulate a variety of physiological processes and represent an important class of drug target. Among the many methods of studying ion channel function, patch clamp electrophysiology is considered the gold standard by providing the ultimate precision and flexibility. However, its utility in ion channel drug discovery is impeded by low throughput. Additionally, characterization of endogenous ion channels in primary cells remains technical challenging. In recent years, many automated patch clamp (APC) platforms have been developed to overcome these challenges, albeit with varying throughput, data quality and success rate. In this study, we utilized SyncroPatch 768PE, one of the latest generation APC platforms which conducts parallel recording from two-384 modules with giga-seal data quality, to push these 2 boundaries. By optimizing various cell patching parameters and a two-step voltage protocol, we developed a high throughput APC assay for the voltage-gated sodium channel Nav1.7. By testing a group of Nav1.7 reference compounds' IC50, this assay was proved to be highly consistent with manual patch clamp (R > 0.9). In a pilot screening of 10,000 compounds, the success rate, defined by > 500 MΩ seal resistance and >500 pA peak current, was 79%. The assay was robust with daily throughput ~ 6,000 data points and Z' factor 0.72. Using the same platform, we also successfully recorded endogenous voltage-gated potassium channel Kv1.3 in primary T cells. Together, our data suggest that SyncroPatch 768PE provides a powerful platform for ion channel research and drug discovery.
Assuntos
Ensaios de Triagem em Larga Escala/métodos , Potenciais da Membrana/fisiologia , Canal de Sódio Disparado por Voltagem NAV1.7/metabolismo , Técnicas de Patch-Clamp/métodos , Bloqueadores dos Canais de Potássio/farmacologia , Bloqueadores dos Canais de Sódio/farmacologia , Animais , Células CHO , Cricetulus , Avaliação Pré-Clínica de Medicamentos , Expressão Gênica , Ensaios de Triagem em Larga Escala/instrumentação , Canal de Potássio Kv1.3/deficiência , Canal de Potássio Kv1.3/genética , Canal de Sódio Disparado por Voltagem NAV1.1/genética , Canal de Sódio Disparado por Voltagem NAV1.1/metabolismo , Canal de Sódio Disparado por Voltagem NAV1.2/genética , Canal de Sódio Disparado por Voltagem NAV1.2/metabolismo , Canal de Sódio Disparado por Voltagem NAV1.3/genética , Canal de Sódio Disparado por Voltagem NAV1.3/metabolismo , Canal de Sódio Disparado por Voltagem NAV1.4/genética , Canal de Sódio Disparado por Voltagem NAV1.4/metabolismo , Canal de Sódio Disparado por Voltagem NAV1.5/genética , Canal de Sódio Disparado por Voltagem NAV1.5/metabolismo , Canal de Sódio Disparado por Voltagem NAV1.6/genética , Canal de Sódio Disparado por Voltagem NAV1.6/metabolismo , Canal de Sódio Disparado por Voltagem NAV1.7/genética , Técnicas de Patch-Clamp/instrumentação , Cultura Primária de Células , Ratos , Canais de Sódio/genética , Canais de Sódio/metabolismo , Linfócitos T/citologia , Linfócitos T/efeitos dos fármacos , Linfócitos T/metabolismo , TransgenesRESUMO
Worldwide heavy oil and bitumen deposits amount to 9 trillion barrels of oil distributed in over 280 basins around the world, with Canada home to oil sands deposits of 1.7 trillion barrels. The global development of this resource and the increase in oil production from oil sands has caused environmental concerns over the presence of toxic compounds in nearby ecosystems and acid deposition. The contribution of oil sands exploration to secondary organic aerosol formation, an important component of atmospheric particulate matter that affects air quality and climate, remains poorly understood. Here we use data from airborne measurements over the Canadian oil sands, laboratory experiments and a box-model study to provide a quantitative assessment of the magnitude of secondary organic aerosol production from oil sands emissions. We find that the evaporation and atmospheric oxidation of low-volatility organic vapours from the mined oil sands material is directly responsible for the majority of the observed secondary organic aerosol mass. The resultant production rates of 45-84 tonnes per day make the oil sands one of the largest sources of anthropogenic secondary organic aerosols in North America. Heavy oil and bitumen account for over ten per cent of global oil production today, and this figure continues to grow. Our findings suggest that the production of the more viscous crude oils could be a large source of secondary organic aerosols in many production and refining regions worldwide, and that such production should be considered when assessing the environmental impacts of current and planned bitumen and heavy oil extraction projects globally.
Assuntos
Aerossóis/análise , Aerossóis/química , Atmosfera/química , Campos de Petróleo e Gás , Indústria de Petróleo e Gás , Alberta , Clima , Atividades Humanas , Hidrocarbonetos/análise , Hidrocarbonetos/química , Material Particulado/análise , Material Particulado/química , Petróleo , VolatilizaçãoRESUMO
BACKGROUND: MicroRNAs (miRNAs), a class of non-coding small RNAs (sRNAs), regulate various biological processes. Although miRNAs have been identified and characterized in several plant species, miRNAs in Asparagus officinalis have not been reported. As a dioecious plant with homomorphic sex chromosomes, asparagus is regarded as an important model system for studying mechanisms of plant sex determination. RESULTS: Two independent sRNA libraries from male and female asparagus plants were sequenced with Illumina sequencing, thereby generating 4.13 and 5.88 million final clean reads, respectively. Both libraries predominantly contained 24-nt sRNAs, followed by 21-nt sRNAs. Further analysis identified 154 conserved miRNAs, which belong to 26 families, and 39 novel miRNA candidates seemed to be specific to asparagus. Comparative profiling revealed that 63 miRNAs exhibited significant differential expression between male and female plants, which was confirmed by real-time quantitative PCR analysis. Among them, 37 miRNAs were significantly up-regulated in the female library, whereas the others were preferentially expressed in the male library. Furthermore, 40 target mRNAs representing 44 conserved and seven novel miRNAs were identified in asparagus through high-throughput degradome sequencing. Functional annotation showed that these target mRNAs were involved in a wide range of developmental and metabolic processes. CONCLUSIONS: We identified a large set of conserved and specific miRNAs and compared their expression levels between male and female asparagus plants. Several asparagus miRNAs, which belong to the miR159, miR167, and miR172 families involved in reproductive organ development, were differentially expressed between male and female plants, as well as during flower development. Consistently, several predicted targets of asparagus miRNAs were associated with floral organ development. These findings suggest the potential roles of miRNAs in sex determination and reproductive developmental processes in asparagus.
Assuntos
Asparagus/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , MicroRNAs/genética , Estabilidade de RNA/genética , RNA de Plantas/genética , Asparagus/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Ontologia Genética , Genes de Plantas/genética , MicroRNAs/metabolismo , Óvulo Vegetal/genética , Óvulo Vegetal/metabolismo , Pólen/genética , Pólen/metabolismo , RNA de Plantas/metabolismo , Reprodução/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
A field study in the Pearl River Delta of China was conducted in order to describe to the spatial and seasonal variation of occurrence and concentrations of total petroleum hydrocarbon (TPH) in surface water and sediments. Petroleum hydrocarbons and isoprenoid alkanes were quantified by UV spectroscopy and gas chromatography with a mass selective detector. The concentrations of TPH ranged from 4.3 to 68.7 µg L(-1) in surface water, and from 66.6 to 1445 µg g(-1) in surface sediments. The ratios of pristine to phytane suggested that the main sources of TPH in the sediment were petroleum importation. The highest concentrations of TPH were present in the spring season. When compared with results from previous studies, it can be concluded that the Pearl River Delta was moderately polluted by TPH. No statistically significant correlations were observed between the concentrations of TPH in surface water and sediments.
Assuntos
Sedimentos Geológicos/química , Hidrocarbonetos/análise , Petróleo/análise , Rios/química , Estações do Ano , Poluentes Químicos da Água/análise , Qualidade da Água , Alcanos/análise , China , Terpenos/análiseRESUMO
Many plants originating from the Asteraceae family are applied as herbal medicines and also beverage ingredients in Asian areas, particularly in China. However, they may be confused due to their similar odor, especially when ground into powder, losing their typical macroscopic characteristics. In this paper, 11 different multiple mathematical algorithms, which are commonly used in data processing, were utilized and compared to analyze the electronic nose (E-nose) response signals of different plants from Asteraceae family. Results demonstrate that three-dimensional plot scatter figure of principal component analysis with less extracted components could offer the identification results more visually; simultaneously, all nine kinds of artificial neural network could give classification accuracies at 100%. This paper presents a rapid, accurate, and effective method to distinguish Asteraceae plants based on their response signals in E-nose. It also gives insights to further studies, such as to find unique sensors that are more sensitive and exclusive to volatile components in Chinese herbal medicines and to improve the identification ability of E-nose. Screening sensors made by other novel materials would be also an interesting way to improve identification capability of E-nose.