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1.
Front Oncol ; 12: 932674, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35928872

RESUMO

Purpose: To seek novel diagnostic approaches, we improved the workflow of cell-free DNA (cfDNA) sequencing and evaluated its feasibility in vitreoretinal lymphoma (VRL) specimens; the profile of mutations was preliminarily analyzed for potential diagnostic value. Methods: The study was a diagnostic trial. 23 eyes of 23 patients with VRL and 25 eyes of 25 patients with inflammatory eye diseases were enrolled. Approximate 500µl undiluted vitreous humor and 10ml diluted vitreous fluid was obtained through diagnostic vitrectomy and sent for cytopathological examinations. 500µl of the diluted vitreous fluid was spared for cfDNA sequencing. For cfDNA sequencing, DNA fragmentation procedure was added to the workflow to improve the extraction efficiency; mutations detected were analyzed for potential diagnostic model. The sensitivity and specificity of the cytopathology and cfDNA sequencing were compared. The clinical manifestations were preliminarily analyzed for potential correlations with the genotypes. Results: CfDNA sequencing was accomplished in 23 eyes with VRL and 20 eyes with inflammatory eye diseases. VRL-related mutated genes included MYD88 (18 eyes, 78%), ETV6 (11 eyes, 48%), PIM1 (11 eyes,48%), BTG2 (7 eyes, 30%), IRF4 (7 eyes, 30%), CD79B (6 eyes, 26%), LRP1B (6 eyes, 26%), etc. Logistic regression based on the mutations of MYD88 and ETV6 was of the potential for the diagnosis of VRL (P<0.001, adjusted R2 = 0.789, sensitivity 0.913, specificity 0.950); by comparison, the sensitivity and specificity of the vitreous cytopathology were 0.826 and 1.000, respectively. Further analysis of the mutation profile showed that patients carrying CD79B mutation tended to have higher intraocular interleukin-10 level (P=0.030), that CARD11 mutation was correlated with younger age at ocular onset (P=0.039), and that patients with intracranial involvement carried more multiple-site mutations in the BTG2 gene (P=0.013). Conclusions: The improved workflow of CfDNA sequencing is of sound feasibility in a limited amount of vitreous humor. The logistic model based on the mutations could help to provide reliable clues for the diagnosis of VRL.

2.
Mini Rev Med Chem ; 22(2): 322-354, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34036917

RESUMO

BACKGROUND: Flavonoid glucuronides are a kind of natural products presenting a flavone linked directly with one or several glucuronides through O-glycoside bond. They had become of interest in natural product research in the past decades for their antioxidant, anti-inflammatory, and antibacteria activities. In particular, the compound breviscapine has a notable effect on cardiocerebrovascular diseases. Several other compounds even have antitumor activity. METHODS: Through searching the database and reading a large number of documents, we summarized the related findings of flavonoid glucuronides. RESULTS: We summarized 211 naturally occurring flavonoid glucuronides in 119 references with their chemical structures, biological activities, and metabolism. A total of 220 references from 1953 to 2020 were cited in this paper according to literature databases such as CNKI, Weipu, Wanfang data, Elsevier, Springer, Wiley, NCBI, PubMed, EmBase, etc. Conclusion: Flavonoid glucuronides are a class of compounds with various chemical structures and a diverse range of biological activities. They are thought to be potential candidates for drug discovery, but the specific study on their mechanisms is still limited until now. We hope this article can provide references for natural product researchers and draw more attention to flavonoid glucuronides' biological activities and mechanisms.


Assuntos
Flavonoides , Glucuronídeos , Anti-Inflamatórios/química , Antioxidantes/química , Antioxidantes/farmacologia , Flavonoides/química , Flavonoides/farmacologia , Glucuronídeos/metabolismo , Glucuronídeos/farmacologia , Compostos Fitoquímicos/química , Extratos Vegetais/química
3.
J Hazard Mater ; 424(Pt A): 127386, 2022 02 15.
Artigo em Inglês | MEDLINE | ID: mdl-34879576

RESUMO

Bis(2ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH), a replacement for restricted flame retardants, has become ubiquitous in the environment. To reveal the neurotoxicity and underlying mechanism of TBPH, we first evaluated its penetrability through the blood-brain barrier (BBB) using hCMEC/D3 cells as in vitro model, and found TBPH had poor penetrability through BBB with a maximum Papp of 14.8 × 10-6 cms-1. Further study using transgenetic zebrafish (Tg flk1: EGFP) as in vivo model confirmed that TBPH could affect the BBB permeability, probably via affecting the transcription of genes encoding tight junction proteins. Finally, wild type zebrafish embryos/larvae were exposed to TBPH to evaluate the neurotoxicity. The neurodevelopment, neurotransmitters and locomotor activity of zebrafish larvae did not changed, which may be because TBPH can hardly cross the BBB to pose direct exposure to the central nervous system. However, the transcription of opsins genes and visual response to light stimulation in zebrafish larvae were inhibited, pointing to additional mechanism that may cause visual impairment indirectly. Above all, these results can help further understand the neurotoxicity and underlying mechanism by TBPH, and also pointed out potential risk of this chemical to aquatic organisms.


Assuntos
Retardadores de Chama , Ácidos Ftálicos , Animais , Barreira Hematoencefálica , Retardadores de Chama/toxicidade , Permeabilidade , Ácidos Ftálicos/toxicidade , Peixe-Zebra
4.
Anim Biotechnol ; : 1-11, 2021 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-34890305

RESUMO

KLF7 belongs to the Krüppel-like factors (KLFs) family, which function as transcriptional regulators controlling a number of basic cellular processes, involving proliferation, differentiation, and migration. Here, we reveal insights into the differentiated expression of KLF7 in different goat tissues and different stages of growth, and the inhibition role of KLF7 knockdown to differentiation by using goat intramuscular and subcutaneous preadipocytes. We demonstrate that KLF7 expression is obviously changed during the differentiation of preadipocytes into mature adipocytes. Knockdown of KLF7 inhibited lipid droplet accumulation, reduced the expression of adipogenic markers both in intramuscular and subcutaneous preadipocytes in goats, suggesting that KLF7 is a novel regulator of adipogenesis. KLF7 expression changed also up or down-regulation the other KLF family members, but there were differences between these two types of cells. Investigation into the mechanism that KLF7 regulates preadipocyte differentiation revealed that KLF family members KLF1, KLF5, KLF6, KLF8, KLF11, KLF12, KLF16, KLF17 and adipogenic markers C/EBPα and SREBP1 promoter region present KLF7 transcriptional binding sites. Altogether, the data here identify KLF7 as a novel regulator of adipogenesis.

5.
Anim Biotechnol ; : 1-13, 2021 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-34939903

RESUMO

Fibroblast growth factor 1(FGF1) has been proved to bind to specific signal molecules and activate intracellular signal transduction, leading to proliferation or differentiation of cells. However, the role of FGF1 in goat adipocytes is still unclear. Here, we investigated its role in lipogenesis of goats, which depends on the activation of FGFRs. In goat intramuscular and subcutaneous adipocytes, we observed that adipocytes accumulation was inhibited by interfering of FGF1, the expression of C/EBPα, C/EBPß, LPL, Pref-1, PPARγ, AP2, KLF4, KLF6, KLF10 and KLF17 were significantly down-regulated (p < 0.05). When the FGF1 was up-regulated, the opposite result was found, while the expression of C/EBPß, LPL, PPARγ, SREBP1, AP2, KLF4, KLF7, KLF15, KLF16 and KLF17 were increased significantly (p < 0.05) in goat intramuscular and subcutaneous adipocytes. The expression level of FGFR1 was significantly and decreased with the interference of FGF1, and increased with the overexpression of FGF1. But in goat subcutaneous adipocytes, only the expression of FGFR2 was consistent with the expression of FGF1. Interference methods confirmed that FGFR1 or FGFR2 and FGF1 have the similarly promoting function in adipocytes differentiation. With the co-transfection technology, we confirmed that FGF1 promoted the differentiation of intramuscular and subcutaneous adipocytes might via FGFR1 or FGFR2, respectively.

6.
Environ Pollut ; 290: 118127, 2021 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-34523534

RESUMO

Multiple environmental stressors, including chemicals termed obesogens, contribute to the susceptibility of organisms to obesity. Bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH), a novel brominated flame retardant, is an environmental contaminant that may disrupt lipid metabolism. However, the risk of TBPH leading to obesity remains unknown. Herein, adult female zebrafish fed a normal-fat diet (NFD) or high-fat diet (HFD) were exposed to 0, 0.02 and 2.0 µM TBPH for 6 weeks. The results showed that chronic TBPH exposure lead to significant weight gain, adipocyte hypertrophy, and subcutaneous fat accumulation, which could be enhanced by HFD feeding. HFD individuals also showed significant visceral fat accumulation. Transcription of the main adipokines regulating lipid metabolism associated with the brain-gut axis were significantly affected by TBPH, especially leptin (brain) and adiponectin (intestine). Additionally, peroxisome proliferator-activated nuclear receptor gamma (PPAR-γ) was significantly upregulated in intestine. TBPH increased the abundance of Firmicutes and Bacteroidetes in the gut microbiota in both NFD and HFD groups, resulting in obesity. Interestingly, population diversity analysis indicated that TBPH alone had a comparable impact on gut microbiota composition to that of HDF controls. Thus, TBPH increased the susceptibility of female zebrafish to obesity by disrupting brain-gut axis regulation and gut microbial composition, leading to enhanced fat accumulation under HFD conditions.


Assuntos
Microbioma Gastrointestinal , Obesidade , Ácidos Ftálicos/efeitos adversos , Peixe-Zebra , Animais , Encéfalo , Dieta Hiperlipídica , Feminino , Obesidade/induzido quimicamente
7.
Am J Chin Med ; 49(6): 1369-1397, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34263720

RESUMO

Breviscapine is one of the extracts of several flavonoids of Erigeron breviscapus. Scutellarin is the main active component of breviscapine, and the qualitative or quantitative criteria as well. Scutellarin and its analogs share a similar skeleton of the flavonoids. Breviscapine has been widely used in the treatment of cerebral infarction and its sequelae, cerebral thrombus, coronary heart disease (CHD), and angina pectoris. Breviscapine has a broad spectrum of pharmacological activities, such as increasing blood flow, improving microcirculation, dilating blood vessels, decreasing blood viscosity, promoting fibrinolysis, inhibiting platelet aggregation, and thrombosis formation, etc. In addition, breviscapine and its analogs have significant value for drug research and development because of the superiority of those significant bioactivities. Furthermore, an increasing number of pharmacokinetic studies have explored the mechanism of scutellarin and its analogs. To provide a comprehensive understanding of the current research on breviscapine, scutellarin, and the analogs, the structural features, distribution situation, preparation method, content determination method, clinical applications, pharmacological action as well as pharmacokinetics are summarized in the present review.


Assuntos
Apigenina , Flavonoides , Glucuronatos , Extratos Vegetais , Apigenina/química , Apigenina/farmacocinética , Apigenina/farmacologia , Flavonoides/química , Flavonoides/farmacocinética , Flavonoides/farmacologia , Glucuronatos/química , Glucuronatos/farmacocinética , Glucuronatos/farmacologia , Humanos , Estrutura Molecular , Extratos Vegetais/química , Extratos Vegetais/farmacocinética , Extratos Vegetais/farmacologia
8.
Sci Total Environ ; 792: 148352, 2021 Oct 20.
Artigo em Inglês | MEDLINE | ID: mdl-34147798

RESUMO

The consumption of pharmaceuticals and personal care products (PPCPs) for controlling and preventing the COVID-19 would have sharply increased during the pandemic. To evaluate their post-pandemic environmental impacts, five categories of drugs were detected in lakes and WWTP-river-estuary system near hospitals of Jinyintan, Huoshenshan and Leishenshan in the three regions (J, H and L) (Regions J, H and L) in Wuhan, China. The total amount of PPCPs (ranging from 2.61 to 1122 ng/L in water and 0.11 to 164 ng/g dry weight in sediments) were comparable to historical reports in Yangtze River basin, whereas the detection frequency and concentrations of ribavirin and azithromycin were higher than those of historical studies. The distribution of concerned drugs varied with space, season, media and water types: sampling sites located at WWTPs-river-estuary system around two hospitals (Regions L and J) usually had relatively high waterborne contamination levels, most of which declined in autumn; lakes had relatively low waterborne contamination levels in summer but increased in autumn. The potential risks of detected PPCPs were further evaluated using the multiple-level ecological risk assessment (MLERA): sulfamethoxazole and azithromycin were found to pose potential risks to aquatic organisms according to a semi-probabilistic approach and classified as priority pollutants based on an optimized risk assessment. In general, the COVID-19 pandemic did not cause serious pollution in lakes and WWTPs-river-estuary system in Wuhan City. However, the increased occurrence of certain drugs and their potential ecological risks need further attention. A strict source control policy and an advanced monitoring and risk warning system for emergency response and long-term risk control of PPCPs is urgent.


Assuntos
COVID-19 , Cosméticos , Preparações Farmacêuticas , Poluentes Químicos da Água , China , Cosméticos/análise , Monitoramento Ambiental , Estuários , Humanos , Lagos , Pandemias , Medição de Risco , Rios , SARS-CoV-2 , Poluentes Químicos da Água/análise
9.
Environ Pollut ; 285: 117471, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34082372

RESUMO

Tris (1,3-dichloro-2-propyl) phosphate (TDCIPP), an alternative to brominated flame retardants, might pose an exposure risk to humans and wild animals during fetal development. Our recent study suggested that short-term TDCIPP exposure during early development caused sex-dependent behavioral alteration in adults. In the present study, multigenerational neurodevelopmental toxicity upon early-life exposure of parental zebrafish was evaluated, and the possible underlying mechanisms were further explored. Specifically, after embryonic exposure (0-10 days post-fertilization, dpf) to TDCIPP (0, 0.01, 0.10, and 1.00 µM), zebrafish larvae were cultured in clean water until the sexually matured to produce progeny (F1). The results confirmed neurodevelopmental toxicity in F1 larvae characterized by changes of developmental endpoints, reduced thigmotaxis, as well as altered transcription of genes including myelin basic protein a (mbpa), growth associated protein (gap43) and synapsin IIa (syn2a). Sex-specific changes in thyroid hormones (THs) indicated the relationship of abnormal THs levels with previously reported neurotoxicity in adult females after early-life exposure to TDCIPP. Similar changing profiles of TH levels (increased T3 and decreased T4) in adult females and F1 eggs, but not in F1 larvae, suggested that the TH disruptions were primarily inherited from the maternal fish. Further results demonstrated hypermethylation of global DNA and key genes related to TH transport including transthyretin (ttr) and solute carrier family 16 member 2 (slc16a2), which might affect the transport of THs to target tissues, thus at least partially contributing to the neurodevelopmental toxicity in F1 larvae. Overall, our results confirmed that early-life TDCIPP exposure of parental fish could affect the early neurodevelopment of F1 offspring. The underlying mechanism could involve altered TH levels inherited from maternal zebrafish and epigenetic modifications in F1 larvae.


Assuntos
Retardadores de Chama , Simportadores , Poluentes Químicos da Água , Animais , Epigênese Genética , Feminino , Retardadores de Chama/toxicidade , Humanos , Larva/metabolismo , Masculino , Transportadores de Ácidos Monocarboxílicos , Compostos Organofosforados , Fosfatos/metabolismo , Hormônios Tireóideos , Poluentes Químicos da Água/toxicidade , Peixe-Zebra/metabolismo
10.
Environ Sci Technol ; 55(10): 6926-6935, 2021 05 18.
Artigo em Inglês | MEDLINE | ID: mdl-33938212

RESUMO

Bis(2-ethylhexyl)-2,3,4,5-tetrabromophthalate (TBPH), a novel brominated flame retardant, can potentially cause lipid metabolism disorder; however, its biological effects on lipid homeostasis remain unknown. We investigated its ability to cause nonalcoholic fatty liver disease (NAFLD) in zebrafish. Female zebrafish were fed a high-fat diet (HFD, 24% crude fat) or normal diet (ND, 6% crude fat), and exposed to TBPH (0.02, 2.0 µM) for 2 weeks. Consequently, HFD-fed fish showed a higher measured concentration of TBPH than ND-fed fish. Further, TBPH-treated fish in the HFD group showed higher hepatic triglyceride levels and steatosis. In comparison to ND-fed fish, treating HFD-fed fish with TBPH led to an increase in the concentration of several proinflammatory markers (e.g., TNF-α, IL-6); TBPH exposure also caused oxidative stress. In addition, the mRNA levels of genes encoding peroxisome proliferator-activated receptors were increased, and the transcription of genes involved in lipid synthesis, transport, and oxidation was upregulated in both ND- and HFD-fed fish. Both the ND and HFD groups also showed demethylation of the peroxisome proliferator-activated receptor-γ coactivator 1-α gene promoter, accompanied by the upregulation of tet1 and tet2 transcription. To summarize, we found that TBPH amplified the disruption of lipid homeostasis in zebrafish, leading to the enhancement of diet-induced NAFLD progression.


Assuntos
Retardadores de Chama , Hepatopatia Gordurosa não Alcoólica , Animais , Feminino , Retardadores de Chama/toxicidade , Homeostase , Fígado , Hepatopatia Gordurosa não Alcoólica/induzido quimicamente , Peixe-Zebra
11.
Front Cell Infect Microbiol ; 11: 639083, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33718285

RESUMO

This study aimed to detetct Mycoplasma bovis (M. bovis) in bovine milk quickly and directly by developing and validating isothermal recombinase polymerase amplification (RPA) assays. Targeting the uvrC gene of M. bovis, an RPA assay based on the fluorescence monitoring (real-time RPA) and an RPA assay combined with a lateral flow strip (LFS RPA) were conducted. It took 20 min for the real-time RPA to finish in a Genie III at 39°C, and 15 min were required to perform the LFS RPA in an incubator block at 39°C, followed by the visualization of the products on the lateral flow strip within 5 min. Both of the two assays showed high specificity for M. bovis without any cross-reaction with the other tested pathogens. With the standard recombinant plasmid pMbovis-uvrC serving as a template, both RPA assays had a limit of detcion of 1.0 × 101 copies per reaction, equivalent to that of a real-time PCR assay. In the 65 milk samples collected from cattle with mastitis, the M. bovis genomic DNA was detected in 24 samples by both the real-time RPA and the LFS RPA assays. The developed RPA assays could detect M. bovis in bovine milk in an efficient, convenient, and credible manner as attractive and promising tools, and the assays would be helpful in the rapid response to M. bovis infection causing bovine mastitis.


Assuntos
Mycoplasma bovis , Animais , Bovinos , Feminino , Leite , Mycoplasma bovis/genética , Técnicas de Amplificação de Ácido Nucleico , Reação em Cadeia da Polimerase em Tempo Real , Recombinases , Sensibilidade e Especificidade
12.
Environ Pollut ; 270: 116277, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33360065

RESUMO

Strobilurin fungicides have been frequently detected in aquatic environments and can induce mitochondrial toxicity to non-target aquatic organisms. However, the derived toxicity and subsequent mechanisms related to their adverse effects are not fully elucidated. In the present study, we compared the mitochondrial and developmental toxicity of azoxystrobin, pyraclostrobin, and trifloxystrobin using zebrafish embryo/larvae. The results showed that all three strobilurins inhibited mitochondrial and non-mitochondrial respiration (the potency is pyraclostrobin ≈ trifloxystrobin > azoxystrobin). Behavioral changes indicated that sublethal doses of pyraclostrobin and azoxystrobin caused hyperactivity of zebrafish larvae in dark cycles, whereas trifloxystrobin resulted in hypoactivity of zebrafish larvae. In addition, pyraclostrobin exposure impaired the inflation of swim bladder, and caused down-regulation of annexin A5 (anxa5) mRNA levels, and up-regulated transcript levels of pre-B-cell leukemia homeobox 1a (pbx1a); conversely, azoxystrobin and trifloxystrobin did not cause detectable effects with swim bladder inflation. Molecular docking results indicated that azoxystrobin had higher interacting potency with iodotyrosine deiodinase (IYD), prolactin receptor (PRLR), antagonistic conformation of thyroid hormone receptor ß (TRß) and glucocorticoid receptor (GR) compared to pyraclostrobin and trifloxystrobin; pyraclostrobin and azoxystrobin were more likely to interact with the antagonistic conformation of TRß and GR, respectively. These results may partially explain the different effects observed in behavior and swim bladder inflation, and also point to potential endocrine disruption induced by these strobilurins. Taken together, our study revealed that all three strobilurins alter mitochondrial bioenergetics and cause developmental toxicity. However, the toxic phenotypes and underlying mechanisms of each chemical may differ, and this requires further investigation. Pyraclostrobin showed higher mitochondrial toxicity at lethal doses and higher developmental toxicity at sublethal doses compared to the two other strobilurins tested. These results provide novel information for toxicological study as well as risk assessment of strobilurin fungicides.


Assuntos
Fungicidas Industriais , Peixe-Zebra , Animais , Fungicidas Industriais/toxicidade , Larva , Simulação de Acoplamento Molecular , Estrobilurinas/toxicidade
13.
Materials (Basel) ; 13(19)2020 Oct 03.
Artigo em Inglês | MEDLINE | ID: mdl-33023004

RESUMO

The microstructure and micro-mechanics around the repaired interface, and the tensile properties of laser additive repaired (LARed) Inconel 625 alloy were investigated. The results showed that the microstructure around the repaired interface was divided into three zones: the substrate zone (SZ), the heat-affected zone (HAZ), and the repaired zone (RZ). The microstructure of the SZ had a typical equiaxed crystal structure, displaying simultaneously precipitated block-shaped MC-type carbides (NbC, TiC), with bimodal sizes of approximately 10 µm and 0.5 µm and an irregularly shaped flocculent Laves phase. Recrystallization occurred in the HAZ, and led to significant grain growth; a portion of the second phase dissolved in the original grain boundaries. In the RZ, there was a columnar crystal structure, and the size increased with increasing deposition thickness. Moreover, the microstructure between the layer interface and layer interior was quite different, presenting an overlapping transition zone (OTZ), in which the dendritic structure coarsened and more Laves phase were precipitated, compared to in the layer interior. The hardness and tensile properties of the LARed samples were equivalent to those of the wrought substrate, which indicates that laser additive repairing (LAR) is a reliable repair solution for damaged and mis-machined components comprising Inconel 625 alloy.

14.
BMC Vet Res ; 16(1): 172, 2020 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-32487081

RESUMO

BACKGROUND: Mycoplasmal pneumonia is an important infectious disease that threatens sheep and goat production worldwide, and Mycoplasma ovipneumoniae is one of major etiological agent causing mycoplasmal pneumonia. Recombinase polymerase amplification (RPA) is an isothermal nucleic acid amplification technique, and RPA-based diagnostic assays have been described for the detection of different types of pathogens. RESULTS: The RPA assays using real-time fluorescence detection (real-time RPA) and lateral flow strip detection (LFS RPA) were developed to detect M. ovipneumoniae targeting a conserved region of the 16S rRNA gene. Real-time RPA was performed in a portable florescence scanner at 39 °C for 20 min. LFS RPA was performed in a portable metal bath incubator at 39 °C for 15 min, and the amplicons were visualized with the naked eyes within 5 min on the lateral flow strip. Both assays were highly specific for M. ovipneumoniae, as there were no cross-reactions with other microorganisms tested, especially the pathogens involved in respiratory complex and other mycoplasmas frequently identified in ruminants. The limit of detection of LFS RPA assay was 1.0 × 101 copies per reaction using a recombinant plasmid containing target gene as template, which is 10 times lower than the limit of detection of the real-time RPA and real-time PCR assays. The RPA assays were further validated on 111 clinical sheep nasal swab and fresh lung samples, and M. ovipneumoniae DNA was detected in 29 samples in the real-time RPA, 31 samples in the LFS RPA and 32 samples in the real-time PCR assay. Compared to real-time PCR, the real-time RPA and LFS RPA showed diagnostic specificity of 100 and 98.73%, diagnostic sensitivity of 90.63 and 93.75%, and a kappa coefficient of 0.932 and 0.934, respectively. CONCLUSIONS: The developed real-time RPA and LFS RPA assays provide the attractive and promising tools for rapid, convenient and reliable detection of M. ovipneumoniae in sheep, especially in resource-limited settings. However, the effectiveness of the developed RPA assays in the detection of M. ovipneumoniae in goats needs to be further validated.


Assuntos
Mycoplasma ovipneumoniae/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/veterinária , Pneumonia por Mycoplasma/diagnóstico , Doenças dos Ovinos/diagnóstico , Animais , Mycoplasma ovipneumoniae/genética , RNA Ribossômico 16S , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Recombinases/metabolismo , Ovinos
15.
Vet Med Sci ; 6(4): 894-900, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32452632

RESUMO

Haemophilus parasuis is the etiological agent of Glässer's disease in swine, which associates with severe economic losses in the swine industry worldwide. A real-time recombinase polymerase amplification assay (real-time RPA) was developed for direct and rapid detection of H. parasuis basing on the translation-initiation factor IF2 (infB) gene. The assay was performed successfully at 39°C for 20 min in Genie III, which is portable and chargeable by battery. The developed assay was highly specific for H. parasuis, and the limit of detection of the assay was 6.0 × 103  fg of H. parasuis genomic DNA, which was the same as that of a real-time PCR developed previously. The assay was further evaluated on 68 pig tissue samples, and 18 (26.5%), 20 (29.4%), and 8 (11.8%) samples were positive for H. parasuis by the real-time RPA, real-time PCR and bacterial isolation, respectively. With the bacteria isolation as the reference method, the real-time RPA showed a diagnostic specificity of 83.33% and a diagnostic sensitivity of 100%. The above data demonstrated the well-potentiality and usefulness of the developed real-time RPA assay in reliable diagnosis of swine Glässer's disease, especially in resource limited settings.


Assuntos
Infecções por Haemophilus/veterinária , Haemophilus parasuis/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/veterinária , Doenças dos Suínos/diagnóstico , Animais , Infecções por Haemophilus/diagnóstico , Infecções por Haemophilus/virologia , Haemophilus parasuis/enzimologia , Técnicas de Amplificação de Ácido Nucleico/métodos , Recombinases/análise , Sus scrofa , Suínos , Doenças dos Suínos/virologia
16.
Materials (Basel) ; 13(3)2020 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-32012825

RESUMO

Zr-based bulk metallic glasses have been attracting tremendous interest of researchers because of their unique combination of mechanical and chemical properties. However, their application is limited as large-scale production is difficult due to the limitation of cooling rate. Recently, additive manufacturing technology has been proposed as a new solution for fabricating bulk metallic glasses without size limitation. In this study, selective laser melting technology was used to prepare Zr60Fe10Cu20Al10 bulk metallic glass. The laser parameters for fabricating full dense amorphous specimens were investigated. The mechanical and corrosion resistance properties of the prepared samples were measured by micro-compression and electrochemical corrosion testing, respectively. Lastly, Zr60Fe10Cu20Al10 bulk metallic glass (BMG) with dispersed nano-crystals was made, and good deformation ability was revealed during micro-compression test. The corrosion resistance decreased a bit due to the crystalline phases. The results provide a promising route for manufacturing large and complex bulk metallic glasses with better mechanical property and acceptable corrosion resistance.

17.
BMC Genomics ; 21(1): 115, 2020 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-32013861

RESUMO

BACKGROUND: Dunaliella salina is a good model organism for studying salt stress. In order to have a global understanding of the expression profiles of Dunaliella salina in response to hypersaline stress, we performed quantitative transcriptomic analysis of Dunaliella salina under hypersaline stress (2.5 M NaCl) of different time duration by the second and third generation sequencing method. RESULTS: Functional enrichment of the up-regulated genes was used to analyze the expression profiles. The enrichment of photosynthesis was observed, accompanied by enrichments of carbon fixation, pigment biosynthetic process and heme biosynthetic process, which also imply the enhancement of photosynthesis. Genes responsible for starch hydrolysis and glycerol synthesis were significantly up-regulated. The enrichment of biosynthesis of unsaturated fatty acids implies the plasma membrane undergoes changes in desaturation pattern. The enrichment of endocytosis implies the degradation of plasma membrane and might help the synthesis of new glycerophospholipid with unsaturated fatty acids. Co-enrichments of protein synthesis and degradation imply a higher protein turnover rate. The enrichments of spliceosome and protein processing in endoplasmic reticulum imply the enhancement of regulations at post-transcriptional and post-translational level. No up-regulation of any Na+ or Cl- channels or transporters was detected, which implies that the extra exclusion of the ions by membrane transporters is possibly not needed. Voltage gated Na+ and Cl- channels, mechanosensitive ion channel are possible signal receptors of salt stress, and Ca2+ and MAP kinase pathways might play a role in signal transduction. CONCLUSION: At global transcriptomic level, the response of Dunaliella salina to hypersaline stress is a systematic work, possibly involving enhancements of photosynthesis, carbon fixation, and heme biosynthetic process, acceleration of protein turnover, spliceosome, protein processing in endoplasmic reticulum, and endocytosis, as well as degradation of starch, synthesis of glycerol, membrane lipid desaturation. Altogether, the changes of these biological processes occurred at trancriptomic level will help understand how a new intracellular balance achieved in Dunaliella salina to adapt to hypersaline environment, which are worth being confirmed at the physiological levels.


Assuntos
Clorófitas/genética , Clorófitas/fisiologia , Estresse Salino/fisiologia , Transcriptoma/genética , Cálcio/metabolismo , Clorófitas/metabolismo , Glicerol/metabolismo , Transdução de Sinais/genética , Sódio/metabolismo , Regulação para Cima/genética
18.
Environ Int ; 134: 105293, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31731001

RESUMO

Early-life exposure to toxicants could affect health outcomes in adulthood. We determined the effects of early-life exposure to the organophosphorus flame-retardant tris (1,3-dichloro-2-propyl) phosphate (TDCIPP) in adult zebrafish. Embryos were exposed to TDCIPP from early embryogenesis (2 h post-fertilization) to 10 days post-fertilization (dpf), and larvae were transferred to clean water until adulthood (150 dpf). TDCIPP showed accumulation in larvae, but returned to control levels after 7 days of depuration. In adult zebrafish exposed to TDCIPP in early life, vulnerability to anxiety-like behavior was observed in females but not males, suggesting gender-dependent neurotoxicity. Decreased dopamine (DA) concentration and down-regulation of dopaminergic signaling related genes were observed in the brains of adult females. Upregulation of DNA methylation transferases (dnmt1, dnmt3a, and dnmt3b) genes were observed in larvae and brains of adult females. Further, the promoter regions of the selected key genes (bdnf, drd4b, zc4h2 and th) showed increased DNA methylation status, accompanied by down-regulation of gene transcription in larvae and brains of adult females. Our results indicate that early-life exposure to TDCIPP could cause delayed neurotoxicity in adult zebrafish.


Assuntos
Metilação de DNA , Animais , Feminino , Retardadores de Chama , Masculino , Organofosfatos , Compostos Organofosforados , Fosfatos , Poluentes Químicos da Água , Peixe-Zebra
19.
Mol Cell Probes ; 45: 14-18, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-30930280

RESUMO

Actinobacillus pleuropneumoniae is the etiological agent of swine contagious pleuropneumoniae, which is distributed globally and associated with severe economic losses in the pig rearing industry. In this study, a real-time recombinase polymerase amplification assay (real-time RPA) based on the apxIVA gene was developed to rapid detect A. pleuropneumoniae. Real-time RPA was performed successfully in Genie III at the constant temperature of 39 °C for 20 min. The developed assay was highly specific for A. pleuropneumoniae, and the sensitivity at 95% probability was 536 fg of A. pleuropneumoniae genomic DNA. The real-time RPA for A. pleuropneumoniae was further evaluated on the 112 clinical swine lung and tonsil samples, and 25 (22.3%), 27 (24.1%), and 12 (10.7%) samples were positive for A. pleuropneumoniae by the real-time RPA, real-time PCR and bacterial isolation, respectively. With a real-time PCR as the reference method, the real-time RPA showed a diagnostic specificity of 98.8%, a diagnostic sensitivity of 88.9%, a positive predicative value of 96.0%, a negative predictive value of 96.5%, and a kappa value of 0.900. The above data demonstrated the well potentiality and usefulness of the developed real-time RPA assay in the reliable detection of A. pleuropneumoniae, especially in resource limited settings.


Assuntos
Infecções por Actinobacillus/diagnóstico , Actinobacillus pleuropneumoniae/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/veterinária , Doenças dos Suínos/virologia , Infecções por Actinobacillus/veterinária , Actinobacillus pleuropneumoniae/genética , Animais , Proteínas de Bactérias/genética , Pulmão/microbiologia , Tonsila Palatina/microbiologia , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Recombinases/metabolismo , Sensibilidade e Especificidade , Suínos
20.
J Microbiol Methods ; 159: 56-61, 2019 04.
Artigo em Inglês | MEDLINE | ID: mdl-30807776

RESUMO

Mycoplasma hyopneumoniae is the etiological agent of swine enzootic pneumonia, which is associated with high economic losses in swine production worldwide. In this study, recombinase polymerase amplification assays using real-time fluorescence detection (real-time RPA) and lateral flow strip detection (LFS RPA) were developed to detect M. hyopneumoniae based on the conserved region of the mhp165 gene. Real-time RPA was performed in Genie III at 39 °C for 20 min, while the LFS RPA was performed in an incubator block at 39 °C for 15 min, and the products were visible on the LFS inspected by the naked eyes within 2 min. Both assays were specific for M. hyopneumoniae, as there were no cross-reactions with other pathogens tested. The limit of detection of both RPA assay was 5.0 × 102 fg of M. hyopneumoniae DNA, which was the same as that of a real-time PCR assay. Of the 146 clinical samples, M. hyopneumoniae DNA was identified in 41, 42, and 47 samples by the real-time RPA, LFS RPA and real-time PCR, respectively. Compared to real-time PCR, the real-time RPA and LFS RPA assays showed diagnostic specificity of 100%, a diagnostic sensitivity of 87.23% and 89.36%, and a kappa value of 0.903 and 0.909, respectively. These results have demonstrated that the developed RPA assays are suitable for rapid and reliable detection of M. hyopneumoniae in diagnostic laboratory and at point-of-need facility.


Assuntos
Mycoplasma hyopneumoniae/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Pneumonia Suína Micoplasmática/microbiologia , Animais , Proteínas de Bactérias/genética , Mycoplasma hyopneumoniae/genética , Recombinases/química , Sensibilidade e Especificidade , Suínos
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