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1.
Magn Reson Imaging ; 69: 57-64, 2020 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-32171775

RESUMO

RATIONALE AND OBJECTIVES: In magnetic resonance (MR) fetal imaging, the image quality acquired by the traditional Cartesian-sampled breath-hold T1-weighted (T1W) sequence may be degraded by motion artifacts arising from both mother and fetus. The radial VIBE sequence is reported to be a viable alternative to conventional Cartesian acquisition for both pediatric and adult MR, yielding better image quality. This study evaluated the role of radial VIBE in fetal MR imaging and compared its image quality and motion artifacts with those of the Cartesian T1W sequence. MATERIALS AND METHODS: We included 246 pregnant women with 50 lesions on 1.5-T MR imaging. Image quality and lesion conspicuity were evaluated by two radiologists, blinded to the acquisition schemes used, using a five-point scale, where a higher score indicated a better trajectory method. Mixed-model analysis of variance and interobserver variability assessment were performed. RESULTS: The radial VIBE sequence showed a significantly better performance than conventional T1W imaging in the head and neck, fetal body, and placenta region: 3.92 ± 0.88 vs 3 ± 0.74, p < 0.001, 3.8 ± 0.94 vs 3.15 ± 0.87, p < 0.001, and 4.17 ± 0.63 vs 3.12 ± 0.72, p < 0.001, respectively. Additionally, fewer motion artifacts were observed in all regions with the radial VIBE sequence (p < 0.01). Of 50 lesions, 49 presented better lesion conspicuity on radial VIBE images than on T1W images (4.34 ± 0.91 vs 3.48 ± 1.46, p < 0.001). CONCLUSION: For fetal imaging, the radial VIBE sequences yielded better image quality and lesion conspicuity, with fewer motion artifacts, than conventional breath-hold Cartesian-sampled T1W sequences.

2.
J Agric Food Chem ; 2020 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-32193930

RESUMO

Metal-organic frameworks (MOFs) are a class of porous materials with versatile properties. In this study, ZIF-8 was employed to establish a two-enzyme system by encapsulating permeabilized Bacillus subtilis cells coated with glucose isomerase. B. subtilis was constructed by introducing the shuttle plasmid PMA5 associated with the over-expression of trehalose synthase. Using this two-enzyme system, trehalose was produced by trehalose synthase, and the by-product glucose was converted to fructose with the help of glucose isomerase. The decrease of glucose production not only relieved the inhibition of the entire reaction chain, but also increased the final yield of trehalose. The highest trehalose production rate reached 67.7%, and remained above 50% after 20 batches. In addition, the toxicity of the ZIF-8 coating for B. subtilis was investigated by fluorescence microscopy, and was found to be negligible. By simulating an extreme environment, the ZIF-8 coating was demonstrated to have a protective effect on the cells and the enzymes. This study provides a theoretical basis for the application of MOFs in the immobilization of microorganisms and enzymes.

3.
J Cell Mol Med ; 2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32129945

RESUMO

Malignant melanoma is one of the most invasive tumours. However, effective therapeutic strategies are limited, and overall survival rates remain low. By utilizing transcriptomic profiling, tissue array and molecular biology, we revealed that two key ubiquitin-specific proteases (USPs), ubiquitin-specific peptidase10 (USP10) and ubiquitin-specific peptidase10 (USP13), were significantly elevated in melanoma at the mRNA and protein levels. Spautin-1 has been reported as a USP10 and USP13 antagonist, and we demonstrated that spautin-1 has potent anti-tumour effects as reflected by MTS and the colony formation assays in various melanoma cell lines without cytotoxic effects in HaCaT and JB6 cell lines. Mechanistically, we identified apoptosis and ROS-mediated DNA damage as critical mechanisms underlying the spautin-1-mediated anti-tumour effect by utilizing transcriptomics, qRT-PCR validation, flow cytometry, Western blotting and immunofluorescence staining. Importantly, by screening spautin-1 with targeted or chemotherapeutic drugs, we showed that spautin-1 exhibited synergy with cisplatin in the treatment of melanoma. Pre-clinically, we demonstrated that spautin-1 significantly attenuated tumour growth in a cell line-derived xenograft mouse model, and its anti-tumour effect was further enhanced by cotreatment with cisplatin. Taken together, our study revealed a novel molecular mechanism of spautin-1 effecting in melanoma and identified a potential therapeutic strategy in treatment of melanoma patients.

4.
Int Urol Nephrol ; 2020 Mar 02.
Artigo em Inglês | MEDLINE | ID: mdl-32124233

RESUMO

OBJECTIVES: We evaluated the clinical manifestations and outcomes of nocardiosis, a rare opportunistic infection that occurs in patients with nephrotic syndrome. METHODS: The records of NS patients with nocardiosis in a single hospital during 2000-2019 were retrieved and studied in detail. RESULTS: Eleven patients were included. The mean time to develop nocardiosis after glucocorticoid therapy was 11.5 ± 14.8 months. Most patients had fever, elevated white blood cell counts and C-reactive protein, whereas procalcitonin levels were normal or slightly elevated in 91% (10/11) patients, except one patient suffered from septic shock. Nine patients were tested for CD4+ T-cell counts; of these, four patients had counts < 200 cells/µL. The most common site of nocardiosis involvement was lung (100%), followed by subcutaneous tissue (72.7%). Radiological findings for lungs in seven cases were characterized by isolated or scattered nodules and masses, usually located subpleural or close to the hilum. Positive smears of Nocardia were detected in 100% of samples of subcutaneous abscess and pleural fluid. Nine patients received oral trimethoprim-sulfamethoxazole, four of which received combined carbapenem, and the remaining two patients received carbapenem monotherapy. The long-term prognosis was excellent, with a treatment success rate of 100% in all patients. CONCLUSIONS: NS patients can develop immunodeficiency after treatment with glucocorticoid and immunosuppressants. In cases where patients develop systemic multiple abscesses, or lung images reveal isolated or scattered nodules and masses that are subpleural or close to the hilum, nocardial infection should be considered. Early diagnosis and specific treatment may improve patient outcomes.

5.
Bioorg Med Chem Lett ; : 127086, 2020 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-32165043

RESUMO

Honokiol, a natural bioactive neolignan isolated from the bark and leaf of Magnolia officinalis and Magnolia obovata, exhibits many important biological properties. In continuation of our interest in discovery of the agrochemicals derived from the natural sources, thirty-seven new 8/8'-alkylthiol-benzoxazole and N-alkyl/sulfonyl-benzoxazolone derivatives of honokiol were prepared and their insecticidal activities were evaluated against the larvae of Mythimna separata Walker and Plutella xylostella Linnaeus. The results showed that eleven derivatives exhibited potent insecticidal activity against M. separata when compared with the positive control. Particularly, compound 5h displayed the most promising insecticidal activity against M. separata with the final mortality rate (FMR) of 58.6%. Meanwhile, compounds 7n (FMR = 65.3%), 7p (FMR = 61.5%), and 8c (FMR = 65.3%) demonstrated a greater insecticidal activity against P. xylostella than toosendanin, a well-known botanical insecticide. Additionally, the preliminary structure-activity relationships (SARs) were also discussed. This study indicates that these honokiol derivatives could be used as leads for the further derivation and development of the potential pesticide candidates for crop protection.

6.
Proc Natl Acad Sci U S A ; 117(11): 6103-6113, 2020 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-32123069

RESUMO

Clinical observation of the association between cancer aggressiveness and embryonic development stage implies the importance of developmental signals in cancer initiation and therapeutic resistance. However, the dynamic gene expression during organogenesis and the master oncofetal drivers are still unclear, which impeded the efficient elimination of poor prognostic tumors, including human hepatocellular carcinoma (HCC). In this study, human embryonic stem cells were induced to differentiate into adult hepatocytes along hepatic lineages to mimic liver development in vitro. Combining transcriptomic data from liver cancer patients with the hepatocyte differentiation model, the active genes derived from different hepatic developmental stages and the tumor tissues were selected. Bioinformatic analysis followed by experimental assays was used to validate the tumor subtype-specific oncofetal signatures and potential therapeutic values. Hierarchical clustering analysis revealed the existence of two subtypes of liver cancer with different oncofetal properties. The gene signatures and their clinical significance were further validated in an independent clinical cohort and The Cancer Genome Atlas database. Upstream activator analysis and functional screening further identified E2F1 and SMAD3 as master transcriptional regulators. Small-molecule inhibitors specifically targeting the oncofetal drivers extensively down-regulated subtype-specific developmental signaling and inhibited tumorigenicity. Liver cancer cells and primary HCC tumors with different oncofetal properties also showed selective vulnerability to their specific inhibitors. Further precise targeting of the tumor initiating steps and driving events according to subtype-specific biomarkers might eliminate tumor progression and provide novel therapeutic strategy.

7.
Bioorg Chem ; 97: 103696, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32135360

RESUMO

Xanthatin, a natural sesquiterpene lactone, occurs as one of the major constituents of Xanthium plants (Compositae) and exhibits many important biological properties. To discover natural products-based pesticides, forty-nine Michael-type thiol/amino adducts of xanthatin were synthesized and characterized, while their pesticidal activities were investigated. Among them, compounds 2c, 2h, 2i, and 2t exhibited more potent antifungal activity against Botrytis cinerea (IC50 = 0.96, 0.38, 6.33, and 7.21 µg/mL, respectively) than xanthatin and the two commercial fungicides. Compounds 2t and 2u displayed broad-spectrum and excellent antifungal effects against all tested phytopathogenic fungi, while their IC50 values ranged from 7.21 to 75.88 µg/mL. Compounds 2a, 2f, 2l, 2m, 2v, 7c, 7e, 7h, 7i, and 7j showed moderate larvicidal activity against Plutella xylostella Linnaeus. Furthermore, compounds 2b, 7g, and 7h demonstrated significant ovicidal activity against P. xylostella with the LC50 values of 14.04, 10.00, and 11.95 mg/L, respectively. These findings suggest that thiol/amino appended in the C-13 position of xanthatin may improve antifungal and ovicidal activities for the derivatives. It was also noticed that the exocyclic double bond of xanthatin is crucial for its larvicidal activity. This work also provides some important hints for further design, synthesis, and structural modification of the xanthanolides sesquiterpene lactones toward development of the new environmentally friendly pesticides for sustainable agricultural production.

8.
Environ Pollut ; 262: 114304, 2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32179214

RESUMO

Strategies to prevent cadmium (Cd) mobilization by crops under salinity conditions differs among distinct genotypes, but the biological mechanisms of Cd accumulation in different genotype crops promoted by salinity have remained scarce. In this study, we investigated the biological mechanisms of Cd accumulation in two quite different amaranth cultivars of low-Cd accumulator Quanhong (QH) and high-Cd accumulator Liuye (LY) in response to salt stress. Transcriptomes analysis was carried out on leaves and roots tissues of LY and QH grown with exchangeable Cd 0.27 mg kg-1 and salinity 3.0 g kg-1 treatment or control conditions, respectively. A total of 3224 differentially expressed genes (DEGs) in LY (1119 in roots, 2105 in leaves) and 848 in QH (207 in roots, 641 in leaves) were identified. Almost in each fold change category (2-25, 25-210, >210), the numbers of DEGs induced by salinity in LY treatments were much more than those in QH treatments, indicating that LY is more salt sensitive. Gene ontology (GO) analysis revealed that salinity stress promoted soil acidification and Cd mobilization in LY treatments through the enhancive expression of genes related to adenine metabolism (84-fold enrichment) and proton pumping ATPase (50-fold enrichment) in roots, and carbohydrate hydrolysis (2.5-fold enrichment) in leaves compared with that of whole genome, respectively. The genes expression of organic acid transporter (ALMT) was promoted by 2.71- to 3.94-fold in roots, facilitating the secretion of organic acids. Salt stress also inhibited the expression of key enzymes related to cell wall biosynthesis in roots, reducing the physical barriers for Cd uptake. All these processes altered in LY were more substantially compared with that of QH, suggesting that salt sensitive cultivars might accumulate more Cd and pose a higher health risk.

9.
Clin Exp Nephrol ; 2020 Mar 22.
Artigo em Inglês | MEDLINE | ID: mdl-32201917

RESUMO

BACKGROUND: To analyze the incidence of early acute kidney injury (AKI) and perioperative factors following hip and knee joint replacement. METHODS: A total of 6281 patients from the department of orthopedics from January 2016 to July 2018 were enrolled, and 1490 patients undergoing hip and knee arthroplasty met the inclusion criteria. The preoperative, intraoperative and postoperative parameters were recorded. The retrospective cohort study was carried out to analyze predictors for AKI and postoperative creatinine elevation following hip and knee joint replacement. RESULTS: Eighty patients (5.4%) met AKI criteria. Age, American Society of Anesthesiologists (ASA) physical status and preoperative diabetes were identified as independent predictors for postoperative AKI in patients undergoing hip and knee arthroplasty (p < 0.05). Age, male, preoperative diabetes, hypertension, and preoperative creatinine were identified as independent predictors for postoperative creatinine elevation (p < 0.05). Patients with AKI were more likely to enter the ICU than non-AKI patients (25% vs 5.6%, p < 0.05). Compared with non-AKI patients, the total hospital stay (16 [11-22] vs 13 [10-16] days) and postoperative hospital stay (11 [8-14] vs 8 [7-11] days) for AKI patients were significantly prolonged (p < 0.05). CONCLUSION: The study shows age, male, preoperative diabetes, hypertension, and preoperative creatinine were independent predictors for postoperative creatinine elevation. In addition, age, ASA physical status and preoperative diabetes are independent predictors for postoperative AKI in patients undergoing hip and knee joint replacement. Postoperative AKI seems to increase ICU admission and significantly prolonged hospital stay.

10.
Viruses ; 12(3)2020 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-32168921

RESUMO

The objective of this study was to investigate the inhibitory effect of chlorogenin 3-O-ß-chacotrioside derivatives against H5N1 subtype of the highly pathogenic avian influenza (HPAI) viruses and its molecular mechanism. A series of novel small molecule pentacyclic triterpene derivatives were designed and synthesized and their antiviral activities on HPAI H5N1 viruses were detected. The results displayed that the derivatives UA-Nu-ph-5, XC-27-1 and XC-27-2 strongly inhibited wild-type A/Duck/Guangdong/212/2004 H5N1 viruses with the IC50 values of 15.59 ± 2.4 µM, 16.83 ± 1.45 µM, and 12.45 ± 2.27 µM, respectively, and had the selectivity index (SI) > 3, which was consistent with the efficacy against A/Thailand/kan353/2004 pseudo-typed viruses. Four dealt patterns were compared via PRNT. The prevention dealt pattern showed the strongest inhibitory effects than other patterns, suggesting that these derivatives act on the entry process at the early stages of H5N1 viral infection, providing protection for cells against infection. Further studies through hemagglutinin inhibition (HI) and neuraminidase inhibitory (NAI) assay confirmed that these derivatives inhibited H5N1 virus replication by interfering with the viral hemagglutinin function. The derivatives could recognize specifically HA protein with binding affinity constant KD values of 2.57 × 10-4 M and 3.67 × 10-4 M. In addition, through site-directed mutagenesis combined with a pseudovirion system, we identified that the high-affinity docking sites underlying interaction were closely associated with amino acid residues I391 and T395 of HA. However, the potential binding sites of the derivatives with HA did not locate at HA1 sialic acids receptor binding domain (RBD). Taken together, these study data manifested that chlorogenin 3-O-ß-chacotrioside derivatives generated antiviral effect against HPAI H5N1 viruses by targeting the hemagglutinin fusion machinery.

11.
Biotechniques ; 2020 Mar 06.
Artigo em Inglês | MEDLINE | ID: mdl-32141763

RESUMO

The establishment of transgenic plants has greatly promoted the progress of plant research. However, traditional selection methods using antibiotics or herbicides may miss any positive transformants with growth defects. Additionally, screening with antibiotics/herbicides requires a huge amount of seeds, sterile work conditions and a large amount of space to germinate plants, making the selection process time- and labor-consuming. In this study, we constructed a novel stable transformation vector, plasmid of OLE1-GFP T-DNA vector (pOGT), which can shorten the steps of cloning foreign genes into expression vectors by using TA cloning. Additionally, selection of transformed seeds with fluorescence overcomes the difficulties of conventional selection with antibiotics/herbicides and simplifies the screening process for transgenic plants.

12.
Redox Biol ; 32: 101479, 2020 Feb 26.
Artigo em Inglês | MEDLINE | ID: mdl-32143149

RESUMO

The incidence and severity of acute kidney injury (AKI) is increased yearly in diabetic patients. Although the mechanisms for this remain unclear, the prevention of AKI in diabetic nephropathy is feasible and of value. As we detected highly activation of TGF-ß/Smad3 signaling in both human biopsy and mouse model of diabetic nephropathy, we hypothesized that Smad3 activation in diabetic kidneys may increase AKI sensitivity. We tested our hypothesis in vitro using TGF-ß type II receptor (TGF-ßRII) disrupted tubular epithelial cells (TECs) and in vivo in mice with streptozotocin (STZ)-induced diabetic nephropathy before the induction of ischemia/reperfusion (I/R) injury. We found that high glucose (HG)-cultured TECs showed increased inflammation, apoptosis and oxidative stress following hypoxia/reoxygenation (H/R) injury. Disruption of TGF-ßRII attenuated cell injury induced by H/R in HG-treated TECs. Consistently, Smad3 knockdown in diabetic kidney attenuated I/R-induced AKI. Mechanistically, Smad3 binds to p53 and enhances p53 activity in cells treated with HG and H/R, which may lead to TECs apoptosis. Additionally, ChIP assay showed that Smad3 bound with the promoter region of NOX4 and induced ROS production and inflammation. In conclusion, our results demonstrate that Smad3 promotes AKI susceptibility in diabetic mice by interacting with p53 and NOX4.

13.
J Cell Mol Med ; 2020 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-32096914

RESUMO

As a main part of pigmentation disorders, skin depigmentation diseases such as vitiligo and achromic naevus are very common and get more attention now. The pathogenesis of depigmentation includes melanocyte dysfunction and loss, which are possibly caused by heredity, autoimmunity and oxidative stress. Among them, oxidative stress plays a key role; however, few clinical treatments can deal with oxidative stress. As reported, Cistanche deserticola polysaccharide (CDP) is an effective antioxidant; based on that, we evaluated its role in melanocyte and further revealed the mechanisms. In this study, we found that CDP could promote melanogenesis in human epidermal melanocytes (HEMs) and mouse melanoma B16F10 cells, it also induced pigmentation in zebrafish. Furthermore, CDP could activate mitogen-activated protein kinase (MAPK) signal pathway, then up-regulated the expression of microphthalmia-associated transcription factor (MITF) and downstream genes TYR, TRP1, TRP2 and RAB27A. Otherwise, we found that CDP could attenuate H2 O2 -induced cytotoxicity and apoptosis in melanocytes. Further evidence revealed that CDP could enhance NRF2/HO-1 antioxidant pathway and scavenge intracellular ROS. In summary, CDP can promote melanogenesis and prevent melanocytes from oxidative stress injury, suggesting that CDP helps maintain the normal status of melanocytes. Thus, CDP may be a novel drug for the treatment of depigmentation diseases.

14.
Mol Med Rep ; 21(3): 1421-1430, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32016458

RESUMO

The occurrence of hyperpigmentation or hypopigmentation after inflammation is a common condition in dermatology and cosmetology. Since the exact mechanism of its occurrence is not yet known, prevention and treatment are troublesome. Previous studies have confirmed that α­melanocyte­stimulating hormone, stem cell factor and other factors can promote melanogenesis­related gene expression through the activation of signaling pathways. Recent studies have revealed that a variety of inflammatory mediators can also participate in the regulation of melanogenesis in melanocytes. In this review, we summarized that interleukin­18, interleukin­33, granulocyte­macrophage colony stimulating factor, interferon­Î³, prostaglandin E2 have the effect of promoting melanogenesis, while interleukin­1, interleukin­4, interleukin­6, interleukin­17 and tumor necrosis factor can inhibit melanogenesis. Further studies have found that these inflammatory factors may activate or inhibit melanogenesis­related signaling pathways (such as protein kinase A and mitogen activated protein kinase) by binding to corresponding receptors, thereby promoting or inhibiting the expression of melanogenesis­related genes and regulating skin pigmentation processes. This suggests that the development of drugs or treatment methods from the perspective of regulating inflammation can provide new ideas and new targets for the treatment of pigmented dermatosis. This review outlines the current understanding of the inflammation factors' roles in melanogenesis.

15.
Redox Biol ; : 101459, 2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-32086008

RESUMO

Phosphatidylinositol-3,4,5-trisphosphate [PI(3,4,5)P3] is a phosphorylated derivative of phosphatidylinositol 4-phosphate [PI(4)P] and phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2], which recruit and activate AKT in the plasma membrane (PM) to promote cellular survival. ORP5 anchors at the endoplasmic reticulum (ER)-PM contact sites and acts as a PI(4)P and PI(4,5)P2/phosphatidylserine (PS) exchanger. Here, a lipidomics analysis of the sensorimotor cortex revealed that transient middle cerebral artery occlusion (tMCAO) disturbs the homeostasis of phosphatidylinositols (PIs) and PS between the PM and ER. Conditional knockout mice showed that ORP5 contributes to this abnormal distribution. Abolishing the ORP5 gene significantly inhibited apoptosis and autophagy. RNA sequencing and RNA pull down analyses confirmed a competing endogenous RNA pathway in which circ_0001449 sponges miR-124-3p and miR-32-5p to promote Osbpl5 translation. Our data showed that circRNA_0001449 regulates membrane homeostasis via ORP5 and is involved in the AKT survival pathway.

16.
Anal Chim Acta ; 1101: 193-198, 2020 Mar 08.
Artigo em Inglês | MEDLINE | ID: mdl-32029111

RESUMO

A protein gel electrophoresis procedure using 0.05% w/v sarkosyl, is reported. The method called 05SAR-PAGE can be used to identify the native masses, dimeric states and modification states of proteins, and also be suitable for pursuing native electroblotting and immunodetection. It has been demonstrated by NMR spectroscopy that 0.05% w/v SAR is much milder than SDS, so it has subtle effects on the native structure of proteins. Therefore, the non-covalent dimerization of PhoBN and PhoRcp can be identified by 05SAR-PAGE which cannot be observed by SDS-PAGE. It has also been demonstrated that 05SAR-PAGE can be used to identify the phosphorylated or methylated proteins. Besides, 05SAR-PAGE shows the advantages of simple operation and low cost, and can be easily adapted to diverse applications.

17.
J Chin Med Assoc ; 83(3): 288-294, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-32080024

RESUMO

BACKGROUND: The aim of this report was to review experience from a single hospital in treating ureteral obstruction related to endometriosis with robot-assisted laparoscopic ureteral reconstruction. METHODS: This retrospective analysis study (Canadian Task Force classification II-3) was conducted at an academic tertiary hospital. Five female patients with hydronephrosis without significant elevation of serum creatinine levels were enrolled. Ureteral endometriosis with obstruction was suspected on radiological images. Previous treatment with double-J stenting with or without medical treatment had failed in all of the patients. We performed robot-assisted laparoscopic segmental resection for ureteral endometriosis and reconstructed the ureter through ureteroureterostomy (RUU) or ureteroneocystostomy (RUC). The involved ureters included left lower ureter in three patients and right lower ureter in two patients. RUU was performed in four patients and RUC in one patient. All of the operations were completed smoothly without complications. RESULTS: All ureteral endometrioses were successfully resected, and follow-up sonography or intravenous pyelography showed resolution of hydronephrosis in all of the patients. CONCLUSION: Our experience proves the feasibility and efficacy of a robot-assisted approach for this rare situation with good outcomes.

18.
Spectrochim Acta A Mol Biomol Spectrosc ; 232: 118152, 2020 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-32088532

RESUMO

This paper, we introduced Sub-terahertz (Sub-THz) technology to identify nematode DNA sequence. First, data mining technology and restriction enzyme digestion were used to cut out two corresponding sequences, each containing about 100 base pairs that could represent the characteristic fragments of Bursaphelenchus xylophilus (Bx) and Bursaphelenchus mucronatus (Bm) rDNA in internal transcribed spacer 1 (ITS1) region. Then, vibration spectra of the two enzyme-cut sequences were measured by Fourier transform infrared spectroscopy (FTIR). Meanwhile, the spectrum was analyzed by molecular dynamics method. It was found that the calculated and experimental spectra of the two enzyme-cut sequences were consistent, although the differences of the sequences could not be well reflected in the spectra. The vibration modes corresponding to diverse absorption peaks in the spectra were quite different, which were closely related to the internal bases sequencing. This can be used as an indicator for identifying Bx and Bm DNA. Moreover, the normal mode analysis (NMA) method was first adopted for spectral attribution analysis of DNA long sequences. Finally, the vibration spectra of shorter sequences predicted by second-order Markov chains and Monte Carlo method were studied. To some extent, the predicted short sequences can represent the complete sequence as the initial calculation structure.

19.
Int J Toxicol ; 39(2): 115-123, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31933405

RESUMO

Azo dyes are used widely as color additives in food, drugs, and cosmetics; hence, there is an increasing concern about their safety and possible health hazards. In the present study, we chose 4 azo dyes tartrazine, Sunset Yellow, amaranth, and Allura red and evaluated their developmental toxicity on zebrafish embryos. At concentration levels of 5 to 50 mM, we found that azo dyes can induce hatching difficulty and developmental abnormalities such as cardiac edema, decreased heart rate, yolk sac edema, and spinal defects including spinal curvature and tail distortion. Exposure to 100 mM of each azo dye was completely embryolethal. The median lethal concentration (LC50), median effective concentration (EC50), and teratogenic index (TI) were calculated for each azo dye at 72 hours postfertilization. For tartrazine, the LC50 was 47.10 mM and EC50 value was at 42.66 mM with TI ratio of 1.10. For Sunset Yellow, the LC50 was 38.93 mM and EC50 value was at 29.81 mM with TI ratio of 1.31. For amaranth, the LC50 was 39.86 mM and EC50 value was at 31.94 mM with TI ratio of 1.25. For Allura red, the LC50 was 47.42 mM and EC50 value was 40.05 mM with TI ratio of 1.18. This study reports the developmental toxicity of azo dyes in zebrafish embryos at concentrations higher than the expected human exposures from consuming food and drugs containing azo dyes.

20.
Biochem Biophys Res Commun ; 523(3): 733-738, 2020 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-31948765

RESUMO

Histidine kinase (HK) of two-component signal transduction system (TCS) is a potential drug target for treating bacterial infections, and most HKs are bifunctional. We have previously identified the HXXXT motif of HK in HisKA subfamily to perform the phosphatase activity, but the specific working mechanism of the threonine is not well understood. In this paper, we use the phosphate group analog BeF3- to capture the enzymatic intermediates between HK853 and RR468 from Thermotoga maritima during dephosphorylation, and demonstrate that the T264 site is essential for populating capable near attack conformers (NAC) between enzyme and substrate to facilitate catalysis. Importantly, mutations at this site can modulate the phosphatase activity of HK. Our results help to understand the TCS signal transduction mechanisms and provide a reference for drug design.

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