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1.
Artigo em Inglês | MEDLINE | ID: mdl-31894386

RESUMO

PURPOSE: This study aimed to determine the molecular features and clinical outcomes of young patients with non-small cell lung cancer (NSCLC) harboring ALK fusion genes. METHODS: We interrogated the genomic profile of 1652 patients with lung cancer who underwent targeted next-generation sequencing to screen for candidate oncogenic drivers using histological specimens acquired from January 2016 to December 2018. RESULTS: ALK fusions were identified in 101 NSCLC patients, and 52 of them were diagnosed before the age of 50 years (52/367, 14.2%). Of the 52 patients with early-onset disease, 22 (42.3%) were male and 43 (82.7%) never smoked; the median patient age was 44 years (range 28-50 years). The most frequently occurring ALK fusion partner was EML4, which was identified in 80.8% (42/52) of young patients. Compared to the older patients, patients with early-onset disease were more likely to harbor EML4-ALK variant 1 (38.5% vs. 14.3%; P = 0.007). We also identified rare ALK fusions, including CHRNA7-ALK, TACR1-ALK, HIP1-ALK, DYSF-ALK and ITGAV-ALK, in patients with early-onset disease, and patients with these fusions responded well to crizotinib treatment. A statistically significant difference was observed in progression-free survival (PFS) between the young patients and older patients who received crizotinib as the first-line therapy (17.5 months vs 9.0 months, P = 0.048). However, the median PFS of young patients harboring concurrent TP53 mutations was only 6.2 months. CONCLUSION: Unique genetic characteristics were found in ALK-rearranged NSCLC patients with early disease onset, and these patients responded better to crizotinib and had longer PFS compared to patients with later disease onset. However, patients with concomitant TP53 mutations may not have a significant response to treatment.

2.
PLoS One ; 15(1): e0227544, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31917827

RESUMO

AIM: A total of 241 patients with chronic HCV infection were recruited to investigate the association between liver fibrosis and PLT counts, as well as with MPV, PDW and P-LCR indices. METHODS: The determination of PLT indices was carried out using a Sysmex XT-1800i automated hematology analyzer. Serological tests for HA, LN, C-IV and PIIINP were performed in 210 patients. The liver stiffness was measured in 69 patients by transient elastography (FibroScan). RESULTS: The analysis showed that the four serum fibrosis markers were negatively correlated with PLT counts, but positively correlated with the MPV, PDW and P-LCR values. Moreover, a similar pattern was found after analyzing the FibroScan measurements, which were negatively correlated with PLT counts, but positively correlated with MPV, PDW and P-LCR values. We subdivided the HCV-infected patients into mild and advanced fibrosis groups. The PLT counts were significantly decreased and the MPV, PDW and P-LCR values were significantly increased in the advanced fibrosis group when compared with the mild fibrosis group. CONCLUSIONS: Our results demonstrate that not only the PLT counts but also the MPV, PDW and P-LCR indices significantly correlate with liver fibrosis in HCV-infected patients. Therefore, these indices may be useful laboratory measures for evaluating liver fibrosis progression.

3.
Oncologist ; 25(1): 15-18, 2020 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31848313

RESUMO

Cervical carcinosarcoma is an extremely rare type of neoplasm that lacks standard of care. Preclinical and clinical evidence has suggested that cryoablation in combination with immunotherapy may result in a synergistic effect, generating a more robust immune response to distant lesions. A few clinical trials have evaluated the efficacy of such combination treatment in a variety of solid tumors, but with conflicting results. This report describes the first clinical efficacy of cryoablation followed by pembrolizumab observed in a patient with tumor mutational burden (TMB)-high metastatic cervical carcinosarcoma that was negative for programmed cell death protein 1 expression, microsatellite instability stable, and had mutations in DNA polymerase epsilon (POLE). She had achieved complete response (CR) after 3 months of pembrolizumab treatment and had maintained CR as of the time of submission of this manuscript, with a progression-free survival of 11 months and counting. The case exhibited an exceptional response to cryoablation followed by pembrolizumab, potentially attributed to mutations in POLE, which lead to an extremely high TMB. This report paves the avenue for establishing treatment regimens for patients with TMB-high cervical carcinosarcoma. KEY POINTS: Owing to its rarity, cervical carcinosarcoma has not been well characterized, and currently, there is no standard of care for this disease. This report describes the first case of clinical efficacy of cryoablation followed by pembrolizumab observed in a patient with tumor mutational burden-high metastatic cervical carcinosarcoma. The case exhibited an exceptional response (maintained CR as of the time of submission of this article: 11 months) to cryoablation followed by pembrolizumab. This is the first POLE-mutated cervical carcinosarcoma case.

4.
Protein Expr Purif ; 168: 105565, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31887428

RESUMO

The present study recombinantly expressed a citrate synthase from cyanobacteria Anabaena sp. PCC7120 (AnCS) in Escherichia coli and characterized its enzymatic activity. The molecular mass of native AnCS was 88,533.1 Da containing two 44,162.7 Da subunits. Recombinant AnCS revealed the highest activity at pH 9.0 and 25 °C. AnCS displayed high thermal stability with a half-life time (t1/2) of approximately 6.5 h at 60 °C, which was more thermostable than most CS from general organisms, but less than those from hyperthermophilic bacteria. The Km values of oxaloacetate and acetyl-CoA were 138.50 and 18.15 µM respectively, suggesting a higher affinity to acetyl-CoA than oxaloacetate. Our inhibition assays showed that AnCS activity was not severely affected by most metal ions, but was strongly inhibited by Cu2+ and Zn2+. Treatments with ATP, ADP, AMP, NADH, and DTT depressed the AnCS activity. Overall, our results provide information on the enzymatic properties of AnCS, which contributes to the basic knowledge on CS selection for industrial utilizations.

5.
Int J Biol Sci ; 15(13): 2798-2814, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31853219

RESUMO

Disequilibrium of CD4+ T-cell subpopulations in peripheral blood (PB) of patients with primary immune thrombocytopenia (ITP) has been well established, whereas the profile of CD4+ T-cell subpopulations in bone marrow (BM) remains elusive. In the present study, the frequencies of T helper 22 (Th22), Th17, Th1, Th2, follicular T helper (Tfh) cells and regulatory T cells (Tregs) as well as their effector cytokines in BM and PB from active ITP patients and healthy controls (HCs) were determined. Results showed that the frequencies of Th22, Th17, Th1, and Tfh cells were significantly higher, but Treg number was remarkably lower in BM from ITP patients than from HCs. In the ITP group, it was notable that the numbers of BM Th22, Th17, Th1, Th2, and Tfh cells were significantly elevated compared with the matched PB counterparts, while Treg number in BM was considerably reduced compared with that in PB. In consistence with the BM Th subset pattern, plasma levels of interleukin (IL)-22, IL-17A, and interferon (INF)-γ in BM from ITP patients were significantly increased compared with that from HCs. Therefore, the balance of CD4+ T-cell subsets was disrupted in both BM and PB of ITP patients, suggesting that this might play important roles in the pathophysiological process of ITP.

6.
FEMS Microbiol Lett ; 366(19)2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31755935

RESUMO

Citrate synthase (CS) is an important enzyme in energy conversion and material circulation, participating in many important biochemical processes. In the present study, CS from Microcystis aeruginosa PCC7806 (MaCS) was cloned and expressed in Escherichia coli Rosetta (DE3). The recombinant MaCS was purified and its enzymological properties were characterized. The results showed that MaCS formed dimers in native status. The optimum temperature and pH of MaCS was 30°C and 8.2, respectively. MaCS displayed relative high thermal stability. Treatment at 50°C for 20 min only decreased 11.30% activity of MaCS and the half-life of MaCS was approximately 35 min at 55°C. The kcat and Km of acetyl-CoA and oxaloacetic acid were 17.133 s-1 (kcat) and 11.62 µM (Km), 24.502 s-1 and 103.00 µM, respectively. MaCS activity was not drastically inhibited by monovalent ions and NADH but depressed by divalent ions and some small molecular compounds, especially Mg2+, Zn2+, Co2+ and DTT. Overall, these data contributed to further understanding of energy metabolism in cyanobacteria and also provided basic information for industrial application of CS.

7.
Ecol Evol ; 9(20): 11647-11656, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31695875

RESUMO

Herbivory tolerance can offset the negative effects of herbivory on plants and plays an important role in both immigration and population establishment. Biomass reallocation is an important potential mechanism of herbivory tolerance. To understand how biomass allocation affects plant herbivory tolerance, it is necessary to distinguish the biomass allocations resulting from environmental gradients or plant growth. There is generally a tight balance between the amounts of biomass invested in different organs, which must be analyzed by means of an allometric model. The allometric exponent is not affected by individual growth and can reflect the changes in biomass allocation patterns of different parts. Therefore, the allometric exponent was chosen to study the relationship between biomass allocation pattern and herbivory tolerance. We selected four species (Wedelia chinensis, Wedelia trilobata, Merremia hederacea, and Mikania micrantha), two of which are invasive species and two of which are accompanying native species, and established three herbivory levels (0%, 25% and 50%) to compare differences in allometry. The biomass allocation in stems was negatively correlated with herbivory tolerance, while that in leaves was positively correlated with herbivory tolerance. Furthermore, the stability of the allometric exponent was related to tolerance, indicating that plants with the ability to maintain their biomass allocation patterns are more tolerant than those without this ability, and the tendency to allocate biomass to leaves rather than to stems or roots helps increase this tolerance. The allometric exponent was used to remove the effects of individual development on allocation pattern, allowing the relationship between biomass allocation and herbivory tolerance to be more accurately explored. This research used an allometric model to fit the nonlinear process of biomass partitioning during the growth and development of plants and provides a new understanding of the relationship between biomass allocation and herbivory tolerance.

8.
J Neuroinflammation ; 16(1): 214, 2019 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-31722723

RESUMO

BACKGROUND: Neonatal hypoxic-ischemic brain damage (HIBD), a leading cause of neonatal mortality, has intractable sequela such as epilepsy that seriously affected the life quality of HIBD survivors. We have previously shown that ion channel dysfunction in the central nervous system played an important role in the process of HIBD-induced epilepsy. Therefore, we continued to validate the underlying mechanisms of TRPV1 as a potential target for epilepsy. METHODS: Neonatal hypoxic ischemia and oxygen-glucose deprivation (OGD) were used to simulate HIBD in vivo and in vitro. Primarily cultured astrocytes were used to assess the expression of TRPV1, glial fibrillary acidic protein (GFAP), cytoskeletal rearrangement, and inflammatory cytokines by using Western blot, q-PCR, and immunofluorescence. Furthermore, brain electrical activity in freely moving mice was recorded by electroencephalography (EEG). TRPV1 current and neuronal excitability were detected by whole-cell patch clamp. RESULTS: Astrocytic TRPV1 translocated to the membrane after OGD. Mechanistically, astrocytic TRPV1 activation increased the inflow of Ca2+, which promoted G-actin polymerized to F-actin, thus promoted astrocyte migration after OGD. Moreover, astrocytic TRPV1 deficiency decreased the production and release of pro-inflammatory cytokines (TNF, IL-6, IL-1ß, and iNOS) after OGD. It could also dramatically attenuate neuronal excitability after OGD and brain electrical activity in HIBD mice. Behavioral testing for seizures after HIBD revealed that TRPV1 knockout mice demonstrated prolonged onset latency, shortened duration, and decreased seizure severity when compared with wild-type mice. CONCLUSIONS: Collectively, TRPV1 promoted astrocyte migration thus helped the infiltration of pro-inflammatory cytokines (TNF, IL-1ß, IL-6, and iNOS) from astrocytes into the vicinity of neurons to promote epilepsy. Our study provides a strong rationale for astrocytic TRPV1 to be a therapeutic target for anti-epileptogenesis after HIBD.

9.
Materials (Basel) ; 12(20)2019 Oct 14.
Artigo em Inglês | MEDLINE | ID: mdl-31614970

RESUMO

As one candidate alloy for future Generation IV and fusion reactors, a dual-phase 12Cr oxide-dispersion-strengthened (ODS) alloy was developed for high temperature strength and creep resistance and has shown good void swelling resistance under high damage self-ion irradiation at high temperature. However, the effect of helium and its combination with radiation damage on oxide dispersoid stability needs to be investigated. In this study, 120 keV energy helium was preloaded into specimens at doses of 1 × 1015 and 1 × 1016 ions/cm2 at room temperature, and 3.5 MeV Fe self-ions were sequentially implanted to reach 100 peak displacement-per-atom at 475 °C. He implantation alone in the control sample did not affect the dispersoid morphology. After Fe ion irradiation, a dramatic increase in density of coherent oxide dispersoids was observed at low He dose, but no such increase was observed at high He dose. The study suggests that helium bubbles act as sinks for nucleation of coherent oxide dispersoids, but dispersoid growth may become difficult if too many sinks are introduced, suggesting that a critical mass of trapping is required for stable dispersoid growth.

10.
Rev Sci Instrum ; 90(9): 093504, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31575228

RESUMO

The development and testing of a Faraday cup fast-ion loss detector capable of measuring sub 100 keV particles is documented. Such measurement capabilities play an important role in the assessment of particle confinement of nuclear fusion experiments. The detector is manufactured using thin-film deposition techniques, building upon previous work using discrete foils. This new manufacturing method allows the form factor of the sensor to become that of essentially a microchip. Analysis of the diagnostic response is performed using Monte-Carlo particle simulations. These simulations show peaks in the detector response at 40 and 70 keV. The sensor is then tested in a tunable linear accelerator capable of accelerating protons from 20 to 120 keV. The detector response was found to be well matched to simulations. Improvements to the design to facilitate robustness are discussed.

11.
PhytoKeys ; 130: 25-32, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31534391

RESUMO

A new species Amentotaxus hekouensis L.M. Gao is described as new to science from Hekou, Yunnan of China, Lao Cai of Vietnam and Xiang Khouang of Laos. The new species is similar to A. argotaenia (Hance) Pilg. in linear or linear-lanceolate leaves, stomatal bands white and microsporophylls 6-8, each with 4-6 pollen sacs, but differs from the latter by its larger leaf size with 8-12.5 cm × 0.9-1.4 cm (vs. 2-11 cm × 0.5-1.1 cm in A. argotaenia), long acuminate leaf apex (vs. rounded to sharply triangular in A. argotaenia), stomatal bands with 25-30 rows (vs. 15-25 rows in A. argotaenia), stomatal bands equal to or slightly narrower than marginal bands (vs. narrower than marginal bands in A. argotaenia); pollen-cone racemes borne 1-2 (vs. 2-4 (10) in A. argotaenia), cones in 12-16 pairs (vs. ca. 12 pairs in A. argotaenia). Its distinctive nature has also been confirmed through DNA barcoding analysis of this genus. The new species is provisionally assessed as endangered (EN) due to its restricted distribution, small population size and the prevalence of habitat destruction within its range.

12.
Int J Ophthalmol ; 12(8): 1317-1322, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31456923

RESUMO

AIM: To investigate whether the gene variants in MYOC and ABCA1 are associated with primary angle-closure glaucoma (PACG) and anterior chamber depth (ACD) and axial length (AL) in samples from northern China. METHODS: The present case-control association study consisted of 500 PACG patients and 720 unrelated controls. Each participant was genotyped for eleven single nucleotide polymorphisms (SNPs) in MYOC and ABCA1 genes (rs12076134, rs183532, rs235875 and rs235913 in MYOC, rs2422493, rs2487042, rs2472496, rs2472493, rs2487032, rs2472459 and rs2472519 near ABCA1) using an improved multiplex ligation detection reaction (iMLDR) technique. The genetic association analyses were performed by PLINK using a logistic regression model. The association between genotypes and ocular biometric parameters was performed by SPSS using generalized estimation equation. Bonferroni corrections were implemented and the statistical power was calculated by the Power and Sample Size Calculation. RESULTS: Two SNPs rs183532 and rs235875 as well as a haplotype TTC in MYOC were nominally associated with PACG despite the significance was lost after Bonferroni correction. No association was observed between ABCA1 and PACG, neither did the association between these variants and ACD as well as AL. CONCLUSION: The present study suggests MYOC and ABCA1 do not play a part in the pathogenesis of PACG as well as the regulation of ocular biometric parameters in a northern Chinese population. Further investigations with large sample size are needed to verify this consequence.

14.
Ecol Lett ; 22(9): 1449-1461, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31267650

RESUMO

The mechanisms underpinning forest biodiversity-ecosystem function relationships remain unresolved. Yet, in heterogeneous forests, ecosystem function of different strata could be associated with traits or evolutionary relationships differently. Here, we integrate phylogenies and traits to evaluate the effects of elevational diversity on above-ground biomass across forest strata and spatial scales. Community-weighted means of height and leaf phosphorous concentration and functional diversity in specific leaf area exhibited positive correlations with tree biomass, suggesting that both positive selection effects and complementarity occur. However, high shrub biomass is associated with greater dissimilarity in seed mass and multidimensional trait space, while species richness or phylogenetic diversity is the most important predictor for herbaceous biomass, indicating that species complementarity is especially important for understory function. The strength of diversity-biomass relationships increases at larger spatial scales. We conclude that strata- and scale- dependent assessments of community structure and function are needed to fully understand how biodiversity influences ecosystem function.


Assuntos
Biodiversidade , Biomassa , Florestas , Plantas/classificação , China , Código de Barras de DNA Taxonômico , Filogenia
15.
Eur J Med Chem ; 179: 723-735, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31284082

RESUMO

The biological resistance of methicillin-resistant staphylococcus aureus (MRSA) has pushed synthetic antibiotics to the forefront. To combat the resistance of MRSA, our new effort directed towards the development of novel structural candidates of enone-bridged indole nitroimidazole scaffolds, and wished to shed some light on the combination of some single pharmacophore with different biological activities. Bioassay revealed that the active compound 4b gave a satisfactory inhibition on MRSA (MIC = 1 µg/mL) and could effectively prevent the development of bacterial resistance. Mechanism exploration indicated that molecule 4b could not only intercalate into MRSA deoxyribonucleic acid (DNA), but also permeate MRSA membrane and bind with penicillin-binding protein 2a (PBP2a), then decreased the expression of three relevant genes in MRSA. Furthermore, it was able to be stored and carried by human serum albumin (HSA), and the participation of metal ions in 4b-HSA system was helpful to improve the supramolecular transport behavior. Hybrid 4b also exhibited low cytotoxicity towards normal lung epithelial cell line BEAS-2B.


Assuntos
Antibacterianos/farmacologia , Regulação Bacteriana da Expressão Gênica/efeitos dos fármacos , Indóis/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Nitroimidazóis/farmacologia , Antibacterianos/síntese química , Antibacterianos/química , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Indóis/química , Staphylococcus aureus Resistente à Meticilina/genética , Testes de Sensibilidade Microbiana , Estrutura Molecular , Nitroimidazóis/química , Relação Estrutura-Atividade
16.
BMC Cancer ; 19(1): 647, 2019 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31262262

RESUMO

BACKGROUND: Recently, long non-coding RNAs (lncRNAs) were considered as important gene expression regulators involving various biological processes. In this study, we explored the role of lncRNAs in the pathogenesis of radiation-induced intestinal fibrosis (RIF). METHODS: LncRNAs were screened by microarray (Human LncRNA Array v3.0, Arraystar, Inc.) and the differentially expressed lncRNAs in RIF and non-RIF were analyzed by bioinformatics methods. The expression of WWC2-AS1/miR-16/FGF2 axis was compared on mRNA and protein level between human intestinal CCD-18Co fibroblasts cell lines and subepithelial SEMFs in response to radiation treatment. The significance of WWC2-AS1 in regulating FGF2 associated proliferation, migration, invasion and fibrosis of CCD-18Co and SEMFs by exposure to radiation was analyzed by shRNA (WWC2-AS1 shRNA) knock-down of endogenous WWC2-AS1. RESULTS: WWC2-AS1 and FGF2 level was significantly higher while miR-16 was down-regulated in radiation-treated intestinal tissues. WWC2-AS1 more potently boosted FGF2 expression via reducing miR-16, and WWC2-AS1 shRNA remarkably inhibited FGF2 associated proliferation, migration, invasion and fibrosis of radiation treatment in vitro, further demonstrating physical interaction between miR-16 and WWC2-AS1 in radiation-induced fibrosis progress. CONCLUSIONS: WWC2-AS1 was highly expressed in RIF, may function as a ceRNA in the regulation of FGF2 by binding miR-16. Targeting WWC2-AS1 thus may benefit radiation-induced fibrosis treatment.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Fator 2 de Crescimento de Fibroblastos/metabolismo , Intestinos/efeitos da radiação , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , RNA Interferente Pequeno/metabolismo , Lesões por Radiação/metabolismo , Linhagem Celular , Colo/metabolismo , Colo/patologia , Colo/efeitos da radiação , Regulação para Baixo , Fibroblastos/metabolismo , Fibrose , Humanos , Intestinos/patologia
17.
Stroke ; 50(9): 2314-2321, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31288671

RESUMO

Background and Purpose- Discrimination of the stability of intracranial aneurysms is critical for determining the treatment strategy, especially in small aneurysms. This study aims to evaluate the feasibility of applying machine learning for predicting aneurysm stability with radiomics-derived morphological features. Methods- Morphological features of 719 aneurysms were extracted from PyRadiomics, of which 420 aneurysms with Maximum3DDiameter ranging from 4 mm to 8 mm were enrolled for analysis. The stability of these aneurysms and other clinical characteristics were reviewed from the medical records. Based on the morphologies with/without clinical features, machine learning models were constructed and compared to define the morphological determinants and screen the optimal model for predicting aneurysm stability. The effect of clinical characteristics on the morphology of unstable aneurysms was analyzed. Results- Twelve morphological features were automatically extracted from PyRadiomics implemented in Python for each aneurysm. Lasso regression defined Flatness as the most important morphological feature to predict aneurysm stability, followed by SphericalDisproportion, Maximum2DDiameterSlice, and SurfaceArea. SurfaceArea (odds ratio [OR], 0.697; 95% CI, 0.476-0.998), SphericalDisproportion (OR, 1.730; 95% CI, 1.143-2.658), Flatness (OR, 0.584; 95% CI, 0.374-0.894), Hyperlipemia (OR, 2.410; 95% CI, 1.029-5.721), Multiplicity (OR, 0.182; 95% CI, 0.082-0.380), Location at middle cerebral artery (OR, 0.359; 95% CI, 0.134-0.902), and internal carotid artery (OR, 0.087; 95% CI, 0.030-0.211) were enrolled into the final prediction model. In terms of performance, the area under curve of the model reached 0.853 (95% CI, 0.767-0.940). For unstable aneurysms, Compactness1 (P=0.035), Compactness2 (P=0.036), Sphericity (P=0.035), and Flatness (P=0.010) were low, whereas SphericalDisproportion (P=0.034) was higher in patients with hypertension. Conclusions- Morphological features extracted from PyRadiomics can be used for aneurysm stratification. Flatness is the most important morphological determinant to predict aneurysm stability. Our model can be used to predict aneurysm stability. Unstable aneurysm is more irregular in patients with hypertension.

18.
Mol Biol Cell ; 30(17): 2296-2308, 2019 08 01.
Artigo em Inglês | MEDLINE | ID: mdl-31166830

RESUMO

In eukaryotic cells, organelle-specific protein quality control (PQC) is critical for maintaining cellular homeostasis. Despite the Golgi apparatus being the major protein processing and sorting site within the secretory pathway, how it contributes to PQC has remained largely unknown. Using different chemical biology-based protein unfolding systems, we reveal the segregation of unfolded proteins from folded proteins in the Golgi. Quality control (QC) substrates are subsequently exported in distinct carriers, which likely contain unfolded proteins as well as highly oligomerized cargo that mimic protein aggregates. At an additional sorting step, oligomerized proteins are committed to lysosomal degradation, while unfolded proteins localize to the endoplasmic reticulum (ER) and associate with chaperones. These results highlight the existence of checkpoints at which QC substrates are selected for Golgi export and lysosomal degradation. Our data also suggest that the steady-state ER localization of misfolded proteins, observed for several disease-causing mutants, may have different origins.

19.
J Vis Exp ; (148)2019 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-31233035

RESUMO

Caenorhabditis elegans (C. elegans) stands out as the only organism in which the challenge of understanding the cellular origins of an entire nervous system can be observed, with single cell resolution, in vivo. Here, we present an integrated protocol for the examination of neurodevelopment in C. elegans embryos. Our protocol combines imaging, lineaging and neuroanatomical tracing of single cells in developing embryos. We achieve long-term, four-dimensional (4D) imaging of living C. elegans embryos with nearly isotropic spatial resolution through the use of Dual-view Inverted Selective Plane Illumination Microscopy (diSPIM). Nuclei and neuronal structures in the nematode embryos are imaged and isotropically fused to yield images with resolution of ~330 nm in all three dimensions. These minute-by-minute high-resolution 4D data sets are then analyzed to correlate definitive cell-lineage identities with gene expression and morphological dynamics at single-cell and subcellular levels of detail. Our protocol is structured to enable modular implementation of each of the described steps and enhance studies on embryogenesis, gene expression, or neurodevelopment.

20.
Eur J Med Chem ; 179: 166-181, 2019 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-31254919

RESUMO

This work did a new exploration towards aminothiazolquinolone oximes as potentially multi-targeting antimicrobial agents. A class of novel hybrids of quinolone, aminothiazole, piperazine and oxime fragments were designed for the first time, conveniently synthesized as well as characterized by 1H NMR, 13C NMR and HRMS spectra. Biological activity showed that some of the synthesized compounds exhibited good antimicrobial activities in comparison with the reference drugs. Especially, O-methyl oxime derivative 10b displayed excellent inhibitory efficacy against MRSA and S. aureus 25923 with MIC values of 0.009 and 0.017 mM, respectively. Further studies indicated that the highly active compound 10b showed low toxicity toward BEAS-2B and A549 cell lines and no obvious propensity to trigger the development of bacterial resistance. Quantum chemical studies have also been conducted and rationally explained the structural features essential for activity. The preliminarily mechanism exploration revealed that compound 10b could not only exert efficient membrane permeability by interfering with the integrity of cells, bind with topoisomerase IV-DNA complex through hydrogen bonds and π-π stacking, but also form a steady biosupramolecular complex by intercalating into DNA to exert the efficient antibacterial activity. The supramolecular interaction between compound 10b and human serum albumin (HSA) was a static quenching, and the binding process was spontaneous, where hydrogen bonds and van der Waals force played vital roles in the supramolecular transportation of the active compound 10b by HSA.


Assuntos
Antibacterianos/farmacologia , Desenho de Drogas , Oximas/farmacologia , Quinolonas/farmacologia , Antibacterianos/síntese química , Antibacterianos/química , DNA/efeitos dos fármacos , DNA Topoisomerase IV/antagonistas & inibidores , DNA Topoisomerase IV/metabolismo , Relação Dose-Resposta a Droga , Humanos , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Testes de Sensibilidade Microbiana , Estrutura Molecular , Oximas/síntese química , Oximas/química , Teoria Quântica , Quinolonas/síntese química , Quinolonas/química , Albumina Sérica Humana/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Relação Estrutura-Atividade
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