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1.
Int J Syst Evol Microbiol ; 70(1): 618-623, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31661045

RESUMO

A golden-pigmented, Gram-strain-negative, aerobic, rod-shaped, non-flagellated and non-gliding bacterium, designated strain lm2T, was isolated from activated sludge obtained from a wastewater treatment plant in Binzhou (Shandong province, PR China). Growth occurred at 15-45°C (optimum, 30 °C), in the presence of 0-5.0 % (w/v) NaCl (optimum, 0-2.0 %) and at pH 6.5-8.0 (optimum, pH 7.0-7.5). The chemotaxonomic, phenotypic and genomic traits were investigated. Phylogenetic analysis based on 16S rRNA gene sequence showed that strain lm2T belonged to the genus Chryseobacterium, with highest sequence similarity to Chryseobacterium echinoideorum CC-CZW010T (97.1 %). Genome sequencing revealed a genome size of 3 611 894 bp and a G+C content of 34.9 mol%. The average nucleotide identity value and the digital DNA-DNA hybridization (dDDH) value between strain lm2T and C. echinoideorum JCM 30470T were 87.8 and 34.7 %, respectively. The major respiratory quinone was Menaquinone-6 (MK-6). The major fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH and iso-C17 : 1 ω9c and its polar lipids consisted of phosphatidylethanolamine (PE), unidentified lipids (L1-5) and unidentified aminolipids (AL1-4). On the basis of these data, strain lm2T is considered to represent a novel species of the genus Chryseobacterium, for which the name Chryseobacterium binzhouense sp. nov. is proposed. The type strain is lm2T (=KCTC 72529T=CCTCC AB2019126T).


Assuntos
Chryseobacterium/classificação , Filogenia , Esgotos/microbiologia , Técnicas de Tipagem Bacteriana , Composição de Bases , China , Chryseobacterium/isolamento & purificação , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfatidiletanolaminas/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Vitamina K 2/análogos & derivados , Vitamina K 2/química , Águas Residuárias
2.
Acta Trop ; 203: 105283, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31811863

RESUMO

Echinococcosis is considered by the World Health Organization (WHO) to be a neglected zoonotic disease in the world. Some Tibetan communities were found to be highly endemic for echinococcosis just 20 years ago. Until recently, we were able to understand the overall disease burden of echinococcosis in Tibetan communities after prevalence data being available from nationwide investigations from 2012 to 2016. Data were abstracted from 9 publications regarding to echinococcosis prevalence between 2016-2018; from 10 data bases on echinococcosis prevalence for 151 Tibetan counties; and statistics of population, Gross Domestic Product (GDP) and health staff from 44 local statistic bureaus and government websites at provincial, prefecture and county level, and 2 books of provincial yearly statistics. These data were used to estimate the Disability Adjusted Life Years (DALYs) due to cystic echinococcosis (CE) and alveolar echinococcosis (AE). The distribution of DALYs was presented geographically and economically. The echinococcosis DALYs in the Tibetan communities were estimated to be 126,159 (95%UI 122,415-137,675) annually using the method recommended by WHO. AE DALYs were estimated to be 105,829 (95%UI 101,969-117,090), which were more than CE DALYs of 20,330 (95%UI 19,690-21,581). Echinococcosis affects people more in underdeveloped areas. There was a tendency that a higher echinococcosis DALYs were usually correlated a higher altitude. Health services are also poorly provided in terms of number of health staff of 5.05 per 1000 population in comparison with the national average of 5.8 per 1000 population. The data suggest that the echinococcosis burden in the center region of Qinghai-Tibet plateau is higher than that of other regions, and consequently more control and health services should be provided to the region.

3.
Sensors (Basel) ; 19(20)2019 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-31614560

RESUMO

Glaucoma is a serious eye disease that can cause permanent blindness and is difficult to diagnose early. Optic disc (OD) and optic cup (OC) play a pivotal role in the screening of glaucoma. Therefore, accurate segmentation of OD and OC from fundus images is a key task in the automatic screening of glaucoma. In this paper, we designed a U-shaped convolutional neural network with multi-scale input and multi-kernel modules (MSMKU) for OD and OC segmentation. Such a design gives MSMKU a rich receptive field and is able to effectively represent multi-scale features. In addition, we designed a mixed maximum loss minimization learning strategy (MMLM) for training the proposed MSMKU. This training strategy can adaptively sort the samples by the loss function and re-weight the samples through data enhancement, thereby synchronously improving the prediction performance of all samples. Experiments show that the proposed method has obtained a state-of-the-art breakthrough result for OD and OC segmentation on the RIM-ONE-V3 and DRISHTI-GS datasets. At the same time, the proposed method achieved satisfactory glaucoma screening performance on the RIM-ONE-V3 and DRISHTI-GS datasets. On datasets with an imbalanced distribution between typical and rare sample images, the proposed method obtained a higher accuracy than existing deep learning methods.

4.
Int J Syst Evol Microbiol ; 69(12): 3830-3836, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31502947

RESUMO

A Gram-stain-negative, non-spore-forming, non-motile, short-rod-shaped and aerobic bacterial strain (designated lm1T) was isolated from propylene oxide saponification wastewater activated sludge obtained from a wastewater treatment facility in Binzhou (Shandong province, PR China). Strain lm1T grew between 15 and 45 °C (optimum, 40 °C). The pH range for growth was at pH 4.0-10.0 (optimum growth at pH 8.0). The range of NaCl concentration for the growth of strain lm1T was 0-4.0 % (w/v), with optimum growth at 1.0-2.0 % (w/v). Phylogenetic analysis based on the 16S rRNA gene sequence showed that strain lm1T belonged to the genus Rhodoligotrophos and was closely related to Rhodoligotrophos appendicifer JCM 16873T (96.7 % 16S rRNA gene sequence similarity) and Rhodoligotrophosjinshengii CCTCC AB2013083T (96.2 %). The average nucleotide identity value and the digital DNA-DNA hybridization value between strain lm1T and R. appendicifer JCM 16873T were 73.4 and 14.3 %, respectively. The major cellular fatty acids of strain lm1T were C19 : 0cyclo ω8c, C18 : 1ω7c and C16 : 0. Strain lm1T contained Q-10 as the predominant respiratory quinone. The polar lipid profile was composed of phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, diphosphatidylglycerol, three unidentified aminolipids, one unidentified glycolipid and five unknown lipids. The G+C content of the genomic DNA was 64.4 mol%. Based on polyphasic taxonomic data, strain lm1T could be classified as a representative of a novel species of the genus Rhodoligotrophos, for which the name Rhodoligotrophosdefluvii sp. nov. is proposed. The type strain is lm1T (=CCTCC AB2019071T=KCTC 72156T).


Assuntos
Alphaproteobacteria/classificação , Filogenia , Esgotos/microbiologia , Alphaproteobacteria/isolamento & purificação , Técnicas de Tipagem Bacteriana , Composição de Bases , China , DNA Bacteriano/genética , Ácidos Graxos/química , Hibridização de Ácido Nucleico , Fosfolipídeos/química , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Ubiquinona/análogos & derivados , Ubiquinona/química , Águas Residuárias/microbiologia
5.
Infect Dis Poverty ; 8(1): 68, 2019 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-31362789

RESUMO

BACKGROUND: Echinococcosis caused by Echinococcus is one of the most major infectious diseases in north-west highland of China. E. granulosus sensu strict, E. multilocularis, and E. canadensis are known to be the only three species related to human health transmitting in the areas. To achieve targeted treatment and control of echinococcosis, the accurate identification and discrimination of the species are important. However, currently the available diagnostic approaches do not present ideal results either in accuracy or efficiency. METHODS: In the study, a set of primers were designed to aim at the three human-pathogenic Echinococcus species in China. The one-step multiplex PCR assay was developed and evaluated for the specificity and sensitivity. A total of 73 parasitic lesions and 41 fecal materials obtained from human and various animals collected in the clinic and the field were tested to assess the applicability of this method. RESULTS: The multiplex PCR effectively detected the individual DNA from the targeted species and their random mixtures generating with distinguishable expected size of products. The detection limit of the assay for each of the three species was 5 pg/µl when they were tested separately. When DNA mixtures of the targeted species containing the same concentration were used as templates, the lowest amount of DNA which can be detected was 50 pg/µl, 10 pg/µl and 5 pg/µl for E. granulosus s. s., E. multilocularis, and E. canadensis respectively. No cross-reactivity was observed when DNA from eight genetically close species was used as control templates. The multiplex PCR identifications of all samples were in line with the original sequencing results except for those infected with E. shiquicus, which showed negative signals in the developed assay. Of all the tested stool materials, 16 were previously found positive for Echinococcus by visual and microscopic examination. Among these 16 samples, 13 were confirmed by the multiplex PCR, and the other three tested negative. Additionally, the multiplex PCR identified another 14 positive feces from the remained 25 stool samples which absence of worms. CONCLUSIONS: The developed multiplex PCR shows advantages in fast diagnosis and large-scale epidemiological investigation, which proven to be a promising tool utilized in clinic and surveillance system.


Assuntos
Equinococose/diagnóstico , Echinococcus/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Animais , China , Diagnóstico Diferencial , Equinococose/classificação , Equinococose/parasitologia , Echinococcus granulosus/isolamento & purificação , Echinococcus multilocularis/isolamento & purificação , Humanos
6.
Structure ; 27(7): 1162-1170.e3, 2019 07 02.
Artigo em Inglês | MEDLINE | ID: mdl-31080119

RESUMO

Arrestins, in addition to desensitizing GPCR-induced G protein activation, also mediate G protein-independent signaling by interacting with various signaling proteins. Among these, arrestins regulate MAPK signal transduction by scaffolding mitogen-activated protein kinase (MAPK) signaling components such as MAPKKK, MAPKK, and MAPK. In this study, we investigated the binding mode and interfaces between arrestin-3 and JNK3 using hydrogen/deuterium exchange mass spectrometry, 19F-NMR, and tryptophan-induced Atto 655 fluorescence-quenching techniques. Results suggested that the ß1 strand of arrestin-3 is the major and potentially only interaction site with JNK3. The results also suggested that C-lobe regions near the activation loop of JNK3 form the potential binding interface, which is variable depending on the ATP binding status. Because the ß1 strand of arrestin-3 is buried by the C-terminal strand in its basal state, C-terminal truncation (i.e., pre-activation) of arrestin-3 facilitates the arrestin-3/JNK3 interaction.


Assuntos
Trifosfato de Adenosina/química , Arrestinas/química , Proteína Quinase 10 Ativada por Mitógeno/química , Trifosfato de Adenosina/metabolismo , Sequência de Aminoácidos , Animais , Arrestinas/genética , Arrestinas/metabolismo , Sítios de Ligação , Clonagem Molecular , Cristalografia por Raios X , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Humanos , Sistema de Sinalização das MAP Quinases/genética , Proteína Quinase 10 Ativada por Mitógeno/genética , Proteína Quinase 10 Ativada por Mitógeno/metabolismo , Modelos Moleculares , Fosforilação , Ligação Proteica , Conformação Proteica em alfa-Hélice , Conformação Proteica em Folha beta , Domínios e Motivos de Interação entre Proteínas , Ratos , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo
7.
J Neurochem ; 148(4): 550-560, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30451284

RESUMO

Protein Phosphatase Mg2+ /Mn2+ -Dependent 1K (PPM1K),also named as PP2Cm or branched-chain α-ketoacid dehydrogenase complex phosphatase, is a member of the metal-dependent phosphatase family and an important metabolic regulator. Single nucleotide polymorphisms (SNPs) in PPM1K contributing to protein functional defects have been found to be associated with numerous human diseases, such as cardiovascular disease, maple syrup urine disease, type 2 diabetes, and neurological disease. PPM1K N94K is an identified missense mutant produced by one of the SNPs in the human PPM1K coding sequence. However, the effects of the N94K mutant on its activity and structural property have not been defined. Here, we performed a detailed enzymological study using steady-state kinetics in the presence of pNPP or phospho-peptide substrates and crystallographic analyses of the wild-type and N94K PPM1K. The PPM1K-N94K significantly impaired its Mg2+ -dependent catalytic activity and structural analysis demonstrated that the N94K mutation induced a conformational change in the key residue in coordinating the Mg2+ in the active site. Specifically, three Mg2+ were located in the active site of the PPM1K N94K instead of two Mg2+ in the PPM1K wild type. Therefore, our results provide a structure basis for the metal ion-dependent PPM1K-N94K phosphatase activity.


Assuntos
Proteína Fosfatase 2C/química , Proteína Fosfatase 2C/genética , Biocatálise , Humanos , Mutação , Relação Estrutura-Atividade
8.
Nat Chem Biol ; 14(9): 876-886, 2018 09.
Artigo em Inglês | MEDLINE | ID: mdl-30120361

RESUMO

Signals from 800 G-protein-coupled receptors (GPCRs) to many SH3 domain-containing proteins (SH3-CPs) regulate important physiological functions. These GPCRs may share a common pathway by signaling to SH3-CPs via agonist-dependent arrestin recruitment rather than through direct interactions. In the present study, 19F-NMR and cellular studies revealed that downstream of GPCR activation engagement of the receptor-phospho-tail with arrestin allosterically regulates the specific conformational states and functional outcomes of remote ß-arrestin 1 proline regions (PRs). The observed NMR chemical shifts of arrestin PRs were consistent with the intrinsic efficacy and specificity of SH3 domain recruitment, which was controlled by defined propagation pathways. Moreover, in vitro reconstitution experiments and biophysical results showed that the receptor-arrestin complex promoted SRC kinase activity through an allosteric mechanism. Thus, allosteric regulation of the conformational states of ß-arrestin 1 PRs by GPCRs and the allosteric activation of downstream effectors by arrestin are two important mechanisms underlying GPCR-to-SH3-CP signaling.


Assuntos
Regulação Alostérica , Arrestina/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , Transdução de Sinais , Domínios de Homologia de src , Células Cultivadas , Células HEK293 , Humanos
9.
Elife ; 72018 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-29393851

RESUMO

Luminal fluid reabsorption plays a fundamental role in male fertility. We demonstrated that the ubiquitous GPCR signaling proteins Gq and ß-arrestin-1 are essential for fluid reabsorption because they mediate coupling between an orphan receptor ADGRG2 (GPR64) and the ion channel CFTR. A reduction in protein level or deficiency of ADGRG2, Gq or ß-arrestin-1 in a mouse model led to an imbalance in pH homeostasis in the efferent ductules due to decreased constitutive CFTR currents. Efferent ductule dysfunction was rescued by the specific activation of another GPCR, AGTR2. Further mechanistic analysis revealed that ß-arrestin-1 acts as a scaffold for ADGRG2/CFTR complex formation in apical membranes, whereas specific residues of ADGRG2 confer coupling specificity for different G protein subtypes, this specificity is critical for male fertility. Therefore, manipulation of the signaling components of the ADGRG2-Gq/ß-arrestin-1/CFTR complex by small molecules may be an effective therapeutic strategy for male infertility.


Assuntos
Regulador de Condutância Transmembrana em Fibrose Cística/metabolismo , Fertilidade , Subunidades alfa Gq-G11 de Proteínas de Ligação ao GTP/metabolismo , Receptores Acoplados a Proteínas-G/metabolismo , beta-Arrestina 1/metabolismo , Animais , Regulador de Condutância Transmembrana em Fibrose Cística/genética , Masculino , Camundongos Endogâmicos C57BL , Camundongos Knockout , Receptores Acoplados a Proteínas-G/genética , beta-Arrestina 1/genética
10.
Ther Drug Monit ; 38(3): 365-70, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-26818624

RESUMO

BACKGROUND: Monohydroxycarbamazepine (MHD, 10-hydroxy-carbamazepine) is the main active metabolite of oxcarbazepine (OXC). The present study aims to investigate the relationship between plasma and saliva concentrations of MHD in Chinese children with epilepsy. METHODS: Plasma and saliva samples were collected and MHD levels were measured by high-performance liquid chromatography system. Linear regression analysis was conducted between the dose of OXC and saliva concentrations, between the dose of OXC and plasma concentrations, and between the saliva concentrations and plasma concentrations. Student's t-test was used for unpaired data. A one-way analysis of variance was used for analyzing co-medication in subgroups of patients. RESULTS: A total of 58 blood samples and 58 saliva samples were obtained from 52 pediatric epileptic patients, with a median age of 5.67 years (0.58-15 years, 23 males and 29 females). There was an apparent positive correlation between the plasma and saliva MHD concentrations [Y = 0.77x - 0.85 (n = 58), R = 0.908, P < 0.01]. MHD plasma and saliva concentrations were positively correlated to daily drug dose (r = 0.461 and 0.417; P < 0.01 respectively). The saliva/plasma MHD ratio was around 0.71 and had no significant difference with age, gender, and combined medications. When data were analyzed for subgroups (one group taking OXC as monotherapy, the second group taking OXC in add-on with non-enzyme-inducing antiepileptic drugs, and the third group taking OXC in add-on with hepatic-enzyme-inducing antiepileptic drugs or moderate inducers), no significant difference was found between plasma and saliva MHD concentrations in all the above 3 groups. CONCLUSIONS: High correlation between plasma and saliva MHD levels supported the use of saliva as an alternative to plasma for OXC monitoring in children with epilepsy.


Assuntos
Anticonvulsivantes/farmacocinética , Carbamazepina/análogos & derivados , Monitoramento de Medicamentos/métodos , Epilepsia/tratamento farmacológico , Adolescente , Anticonvulsivantes/administração & dosagem , Grupo com Ancestrais do Continente Asiático , Carbamazepina/administração & dosagem , Carbamazepina/farmacocinética , Criança , Pré-Escolar , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Feminino , Humanos , Lactente , Modelos Lineares , Masculino , Oxcarbazepina , Estudos Prospectivos , Saliva/química
11.
J Int Med Res ; 41(4): 1318-25, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23812114

RESUMO

OBJECTIVE: To explore the effects of progesterone (P) elevation on pregnancy outcomes of day 3 embryo and day 5 blastocyst transfer. METHODS: Clinical outcomes (pregnancy and ectopic pregnancy rates) following day 3 embryo and day 5 blastocyst transfer cycles were retrospectively analysed. Day 3 embryo and day 5 blastocyst transfer cycles were divided into normal P level (P ≤ 1.5 ng/ml) and P elevation group (P > 1.5 ng/ml), based on the serum P level on the day of human chorionic gonadotropin (hCG) administration. RESULTS: A total of 2868 cycles were analysed. In day 3 embryo transfer cycles (n = 2345), the clinical pregnancy rate was significantly higher in the normal P level group compared with the P elevation group (55.4% versus 46.7%, respectively) and the ectopic pregnancy rate was significantly lower in the normal P level group compared with the P elevation group (2.8% versus 7.9%, respectively). In day 5 blastocyst transfer cycles (n = 523), there were no significant differences in the clinical pregnancy and ectopic pregnancy rates between the two groups, based on the P level. CONCLUSION: These preliminary findings suggest that day 5 blastocyst transfer should be adopted for patients with P elevation on the day of hCG administration.


Assuntos
Blastocisto/fisiologia , Gonadotropina Coriônica/uso terapêutico , Transferência Embrionária/métodos , Fertilização In Vitro , Infertilidade Feminina/terapia , Progesterona/sangue , Adulto , Feminino , Humanos , Infertilidade Feminina/sangue , Infertilidade Feminina/diagnóstico por imagem , Gravidez , Resultado da Gravidez , Taxa de Gravidez , Gravidez Ectópica/diagnóstico por imagem , Gravidez Ectópica/fisiopatologia , Estudos Retrospectivos , Fatores de Tempo , Ultrassonografia
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