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1.
An Acad Bras Cienc ; 93(1): e20200129, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-33852715

RESUMEN

Eragrostis plana (Nees) (Tough Lovegrass) shows ability to interfere with other plants, a phenomenon known as allelopathy. This chemical interaction between plants occurs due to the release of compounds into the environment. Thus, a phytotoxicity study was carried out with E. plana roots collected during each season throughout the year, and the compounds were extracted with solvents of increasing polarity. The data from the bioassays were analyzed by GLM and PCA. In addition, a fingerprint of these extracts was obtained by HPLC-DAD. The extracts in petroleum ether from roots collected in the winter and summer showed greater phytotoxicity on Ipomoea grandifolia germination and growth. The PCA obtained from the chromatogram of the crude extract showed that the extracts in petroleum ether were chemically different from the extracts in ethyl acetate and methanol. Thus, continuing this study in order to develop a new generation of bio-herbicides is essential.


Asunto(s)
Eragrostis , Alelopatía , Bioensayo , Cromatografía Líquida de Alta Presión , Extractos Vegetales/toxicidad , Raíces de Plantas
2.
J Vis Exp ; (168)2021 02 14.
Artículo en Inglés | MEDLINE | ID: mdl-33645588

RESUMEN

Microglia orchestrate neuroimmune responses in several neurodegenerative diseases, including Parkinson's disease and Alzheimer's disease. Microglia clear up dead and dying neurons through the process of efferocytosis, a specialized form of phagocytosis. The phagocytosis function can be disrupted by environmental or genetic risk factors that affect microglia. This paper presents a rapid and simple in vitro microscopy protocol for studying microglial efferocytosis in an induced pluripotent stem cell (iPSC) model of microglia, using a human neuroblastoma cell line (SH-SY5Y) labeled with a pH-sensitive dye for the phagocytic cargo. The procedure results in a high yield of dead neuroblastoma cells, which display surface phosphatidylserine, recognized as an "eat-me" signal by phagocytes. The 96-well plate assay is suitable for live-cell time-lapse imaging, or the plate can be successfully fixed prior to further processing and quantified by high-content microscopy. Fixed-cell high-content microscopy enables the assay to be scaled up for screening of small molecule inhibitors or assessing the phagocytic function of genetic variant iPSC lines. While this assay was developed to study phagocytosis of whole dead neuroblastoma cells by iPSC-macrophages, the assay can be easily adapted for other cargoes relevant to neurodegenerative diseases, such as synaptosomes and myelin, and other phagocytic cell types.


Asunto(s)
Bioensayo/métodos , Células Madre Pluripotentes Inducidas/metabolismo , Macrófagos/metabolismo , Neuroblastoma/patología , Fagocitosis , Animales , Muerte Celular , Línea Celular Tumoral , Análisis de Datos , Colorantes Fluorescentes/química , Células Madre Embrionarias Humanas/citología , Humanos , Concentración de Iones de Hidrógeno , Células Madre Pluripotentes Inducidas/citología , Control de Calidad , Reproducibilidad de los Resultados , Imagen de Lapso de Tiempo
3.
Sci Total Environ ; 771: 144514, 2021 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-33736142

RESUMEN

Every year thousands of chemicals get discharged into the waterbodies of the world. These chemicals cause endocrine disruption and induce adverse health effects in human and aquatic life. Global environmental protection agencies emphasise the need to develop rapid and specific tests for identification of these endocrine disruptive chemicals (EDCs) in water. Detection of chemicals that disrupt androgen signaling is especially important because androgen input at specific phases of life is critical for proper male development. Effect-based methods such as reporter assays are suitable tools for identification of EDCs in mixtures of unknown composition. The current study describes a stable, secreted alkaline protease (SEAP)-based reporter assay system, for visual detection of androgenic/antiandrogenic activity present in water samples. A novel feature of this system is the inclusion of coactivators, GRIP1, CARM1, p300 and mZac1b, in addition to an optimal combination of androgen response element (3× HRE), androgen receptor (AR) and the SEAP reporter gene. Incorporation of the coactivators resulted in a transcriptional fold change of 162 folds, enabling visual detection at much lower concentrations of androgen (1 picomolar) within 1 h of addition of test sample. Also, non-androgenic steroids such as estrogen, progesterone and Dexamethasone did not induce significant reporter activity, except at very high concentrations. This reporter assay can be readily converted into a high throughput format for investigation in multiple samples simultaneously, and reflects the changes that can be expected to occur inside a mammalian cell. The androgenic activity in six different water sources was evaluated using this assay. The results reveal significant androgenic activity in rivers and lakes close to Industrial areas, whereas the highest androgenic activity was observed in water containing paper and pulp mill effluents. This bioassay therefore provides a rapid, visual detection tool for effect-directed analysis of androgenic/antiandrogenic compounds in water. IMPACT STATEMENT: The current SEAP-based assay allows visual detection of androgens/antiandrogens in water, at concentrations as low as 1 picomolar, within a 1 h time period, in a high throughput format, providing a very useful technique for field users and regulatory bodies.


Asunto(s)
Antagonistas de Andrógenos , Andrógenos , Antagonistas de Andrógenos/toxicidad , Animales , Proteínas Bacterianas , Bioensayo , Endopeptidasas , Genes Reporteros , Humanos , Masculino , Agua
4.
Sci Total Environ ; 769: 145218, 2021 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-33736268

RESUMEN

Bisphenol A (BPA) is a well-known xenoestrogen, and UV/H2O2 advanced oxidation process (AOP) is one of the most effective technologies to remove BPA from water. Using BPA spiked tap water, a batch-scale photochemical experiment was conducted to investigate whether BPA can pose a genotoxicity concern during the medium pressure (MP) UV/H2O2 treatment and the post-chloramination. Samples at different UV exposure and post-chloramination durations were collected and analyzed by CALUX® gene reporter assays regarding estrogen receptor α (ERα) and p53 transcriptional activity. MP UV/H2O2 process did not cause extra estrogenic effects from the degradation of BPA, whereas genotoxicity occurred when the treated water was exposed with monochloramine. Seven frequently reported nitrogenous disinfection byproducts (N-DBPs) were detected, but none of them were responsible for the observed genotoxicity. Employed with gas chromatography-quadrupole time-of-flight mass spectrometry (GC-QTOF-MS), four compounds possibly contributed to the genotoxicity were tentatively identified and two of them with aminooxy- or cyano- group were considered as "new" N-DBPs. This study demonstrated that by-products differ from their parent compounds in toxicity can be formed in the UV oxidation with post-disinfection process, which should become a cause for concern.


Asunto(s)
Contaminantes Químicos del Agua , Purificación del Agua , Compuestos de Bencidrilo , Bioensayo , Desinfección , Peróxido de Hidrógeno , Fenoles , Rayos Ultravioleta , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad
5.
Sci Total Environ ; 768: 144750, 2021 May 10.
Artículo en Inglés | MEDLINE | ID: mdl-33736315

RESUMEN

Recent urban public water supply contamination events emphasize the importance of screening treated drinking water quality after distribution. In vitro bioassays, when run concurrently with analytical chemistry methods, are effective tools to evaluating the efficacy of water treatment processes and water quality. We tested 49 water samples representing the Chicago Department of Water Management service areas for estrogen, (anti)androgen, glucocorticoid receptor-activating contaminants and cytotoxicity. We present a tiered screening approach suitable to samples with anticipated low-level activity and initially tested all extracts for statistically identifiable endocrine activity; performing a secondary dilution-response analysis to determine sample EC50 and biological equivalency values (BioEq). Estrogenic activity was detected in untreated Lake Michigan intake water samples using mammalian (5/49; median: 0.21 ng E2Eq/L) and yeast cell (5/49; 1.78 ng E2Eq/L) bioassays. A highly sensitive (anti)androgenic activity bioassay was applied for the first time to water quality screening and androgenic activity was detected in untreated intake and treated pre-distribution samples (4/49; 0.93 ng DHTEq/L). No activity was identified above method detection limits in the yeast androgenic, mammalian anti-androgenic, and both glucocorticoid bioassays. Known estrogen receptor agonists were detected using HPLC/MS-MS (estrone: 0.72-1.4 ng/L; 17α-estradiol: 1.3-1.5 ng/L; 17ß-estradiol: 1.4 ng/L; equol: 8.8 ng/L), however occurrence did not correlate with estrogenic bioassay results. Many studies have applied bioassays to water quality monitoring using only relatively small samples sets often collected from surface and/or wastewater effluent. However, to realistically adapt these tools to treated water quality monitoring, water quality managers must have the capacity to screen potentially hundreds of samples in short timeframes. Therefore, we provided a tiered screening model that increased sample screening speed, without sacrificing statistical stringency, and detected estrogenic and androgenic activity only in pre-distribution Chicago area samples.


Asunto(s)
Disruptores Endocrinos , Contaminantes Químicos del Agua , Animales , Bioensayo , Chicago , Disruptores Endocrinos/análisis , Monitoreo del Ambiente , Estrógenos/análisis , Michigan , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad , Calidad del Agua
6.
Sci Total Environ ; 770: 144748, 2021 May 20.
Artículo en Inglés | MEDLINE | ID: mdl-33736394

RESUMEN

N-nitrosodimethylamine (NDMA) is one of the emerging nitrogenous disinfection by-products (DBPs) with probable cytotoxicity, genotoxicity, and carcinogenesis. Its potential toxicological effects have drawn considerable attention but remain inadequately known. Here two methods, including developmental toxicity evaluation and neurobehavioral toxicity evaluation, were used and compared to investigate the safety of low-dose (0, 0.1, 1, 10, 100 µg/L) NDMA exposure to embryo-larval stage of zebrafish. Results showed that the survival and malformation rate of larvae at 6-day post fertilization (dpf) and the hatching rates at 48-h post fertilization (hpf) and 72 hpf in treatment groups had no significant difference with the control group, indicating that the tested NDMA doses were not developmentally toxic. Differently, the changes of neurobehavioral indicators performed more sensitivity. For example, 100 µg/L NDMA exposure induced locomotor hyperactivity at 7 dpf and induced an increasing effect on the relative path angle value. Further, relative value of path angle had more consistency with locomotion results compared with absolute value, indicating that relative value of path angle may be more suitable for the safety evaluation of low-dose NDMA exposure. This work supported that zebrafish neurobehavioral test is a powerful tool in the safety assessment of low-dose NDMA exposure and also has the potential to evaluate the safety of other DBPs.


Asunto(s)
Dimetilnitrosamina , Pez Cebra , Animales , Bioensayo , Desinfección , Embrión no Mamífero , Larva
7.
Molecules ; 26(4)2021 Feb 16.
Artículo en Inglés | MEDLINE | ID: mdl-33669256

RESUMEN

O-GlcNAcylation is a posttranslational modification that occurs at serine and threonine residues of protein substrates by the addition of O-linked ß-d-N-acetylglucosamine (GlcNAc) moiety. Two enzymes are involved in this modification: O-GlcNac transferase (OGT), which attaches the GlcNAc residue to the protein substrate, and O-GlcNAcase (OGA), which removes it. This biological balance is important for many biological processes, such as protein expression, cell apoptosis, and regulation of enzyme activity. The extent of this modification has sparked interest in the medical community to explore OGA and OGT as therapeutic targets, particularly in degenerative diseases. While some OGA inhibitors are already in phase 1 clinical trials for the treatment of Alzheimer's disease, OGT inhibitors still have a long way to go. Due to complex expression and instability, the discovery of potent OGT inhibitors is challenging. Over the years, the field has grappled with this problem, and scientists have developed a number of techniques and assays. In this review, we aim to highlight assays and techniques for OGT inhibitor discovery, evaluate their strength for the field, and give us direction for future bioassay methods.


Asunto(s)
Bioensayo/métodos , N-Acetilglucosaminiltransferasas/metabolismo , Acetilglucosamina/química , Acetilglucosamina/metabolismo , Fenómenos Biofísicos , Química Clic , Unión Proteica
8.
Molecules ; 26(4)2021 Feb 16.
Artículo en Inglés | MEDLINE | ID: mdl-33669312

RESUMEN

Diabetes mellitus is a chronic disease and one of the fastest-growing health challenges of the last decades. Studies have shown that chronic low-grade inflammation and activation of the innate immune system are intimately involved in type 2 diabetes pathogenesis. Momordica charantia L. fruits are used in traditional medicine to manage diabetes. Herein, we report the purification of a new 23-O-ß-d-allopyranosyl-5ß,19-epoxycucurbitane-6,24-diene triterpene (charantoside XV, 6) along with 25ξ-isopropenylchole-5(6)-ene-3-O-ß-d-glucopyranoside (1), karaviloside VI (2), karaviloside VIII (3), momordicoside L (4), momordicoside A (5) and kuguaglycoside C (7) from an Indian cultivar of Momordica charantia. At 50 µM compounds, 2-6 differentially affected the expression of pro-inflammatory markers IL-6, TNF-α, and iNOS, and mitochondrial marker COX-2. Compounds tested for the inhibition of α-amylase and α-glucosidase enzymes at 0.87 mM and 1.33 mM, respectively. Compounds showed similar α-amylase inhibitory activity than acarbose (0.13 mM) of control (68.0-76.6%). Karaviloside VIII (56.5%) was the most active compound in the α-glucosidase assay, followed by karaviloside VI (40.3%), while momordicoside L (23.7%), A (33.5%), and charantoside XV (23.9%) were the least active compounds. To better understand the mode of binding of cucurbitane-triterpenes to these enzymes, in silico docking of the isolated compounds was evaluated with α-amylase and α-glucosidase.


Asunto(s)
Antiinflamatorios/farmacología , Simulación por Computador , Frutas/química , Glicósidos/química , Glicósidos/farmacología , Hipoglucemiantes/farmacología , Momordica charantia/química , Triterpenos/química , Triterpenos/farmacología , Animales , Antiinflamatorios/química , Bioensayo , Espectroscopía de Resonancia Magnética con Carbono-13 , Glicósidos/aislamiento & purificación , Hipoglucemiantes/química , Ligandos , Ratones , Conformación Molecular , Simulación del Acoplamiento Molecular , Espectroscopía de Protones por Resonancia Magnética , Células RAW 264.7 , ARN Mensajero/genética , ARN Mensajero/metabolismo , Triterpenos/aislamiento & purificación , alfa-Amilasas/química , alfa-Amilasas/metabolismo , alfa-Glucosidasas/química , alfa-Glucosidasas/metabolismo
9.
Molecules ; 26(4)2021 Feb 16.
Artículo en Inglés | MEDLINE | ID: mdl-33669321

RESUMEN

This study was aimed at preparing and characterizing solid lipid nanoparticles loading rutin (RT-SLNs) for the treatment of oxidative stress-induced diseases. Phospholipon 80H® as a solid lipid and Polysorbate 80 as surfactant were used for the SLNs preparation, using the solvent emulsification/diffusion method. We obtained spherical RT-SLNs with low sizes, ranging from 40 to 60 nm (hydrodynamic radius) for the SLNs prepared starting from 2% and 5% (w/w) theoretical amount. All prepared formulations showed negative zeta-potential values. RT was efficiently encapsulated within SLNs, obtaining high encapsulation efficiency and drug content percentages, particularly for SLNs prepared with a 5% theoretical amount of RT. In vitro release profiles and analysis of the obtained data applying different kinetic models revealed Fickian diffusion as the main mechanism of RT release from the SLNs. The morphology of RT-SLNs was characterized by scanning electron microscopy (SEM), whereas the interactions between RT and the lipid matrix were investigated by Raman spectroscopy, evidencing spectral modifications of characteristic bands of RT due to the establishment of new interactions. Finally, antioxidant activity assay on human glioblastoma astrocytoma (U373) culture cells showed a dose-dependent activity for RT-SLNs, particularly at the highest assayed dose (50 µM), whereas the free drug showed the lesser activity.


Asunto(s)
Lípidos/química , Nanopartículas/química , Rutina/farmacología , Antioxidantes/farmacología , Bioensayo , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Liberación de Fármacos , Humanos , Hidrodinámica , Nanopartículas/ultraestructura , Análisis de Regresión , Espectrometría Raman , Electricidad Estática
10.
Molecules ; 26(4)2021 Feb 19.
Artículo en Inglés | MEDLINE | ID: mdl-33669666

RESUMEN

Preliminary bioassay-guided fractionation was performed to identify cytotoxic compounds from Hechtia glomerata, a plant that is used in Mexican ethnomedicine. Organic and aqueous extracts were prepared from H. glomerata's leaves and evaluated against two cancer cell lines. The CHCl3/MeOH (1:1) active extract was fractionated, and the resulting fractions were assayed against prostate adenocarcinoma PC3 and breast adenocarcinoma MCF7 cell lines. Active fraction 4 was further analyzed by high-performance liquid chromatography-quadrupole time-of-flight-mass spectrometry analysis to identify its active constituents. Among the compounds that were responsible for the cytotoxic effects of this fraction were flavonoids, phenolic acids, and aromatic compounds, of which p-coumaric acid (p-CA) and its derivatives were abundant. To understand the mechanisms that underlie p-CA cytotoxicity, a microarray assay was performed on PC3 cells that were treated or not with this compound. The results showed that mitogen-activated protein kinases (MAPKs) that regulate many cancer-related pathways were targeted by p-CA, which could be related to the reported effects of reactive oxygen species (ROS). A molecular docking study of p-CA showed that this phenolic acid targeted these protein active sites (MAPK8 and Serine/Threonine protein kinase 3) at the same binding site as their inhibitors. Thus, we hypothesize that p-CA produces ROS, directly affects the MAPK signaling pathway, and consequently causes apoptosis, among other effects. Additionally, p-CA could be used as a platform for the design of new MAPK inhibitors and re-sensitizing agents for resistant cancers.


Asunto(s)
Bromeliaceae/química , Ácidos Cumáricos/farmacología , Proteínas Quinasas Activadas por Mitógenos/antagonistas & inhibidores , Extractos Vegetales/química , Inhibidores de Proteínas Quinasas/farmacología , Bioensayo , Muerte Celular/efectos de los fármacos , Cromatografía Líquida de Alta Presión , Ácidos Cumáricos/química , Regulación hacia Abajo/efectos de los fármacos , Regulación hacia Abajo/genética , Humanos , Células MCF-7 , Proteínas Quinasas Activadas por Mitógenos/química , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Simulación del Acoplamiento Molecular , Simulación de Dinámica Molecular , Células PC-3 , Fenoles/farmacología , Regulación hacia Arriba/efectos de los fármacos , Regulación hacia Arriba/genética
11.
Mikrochim Acta ; 188(4): 137, 2021 03 25.
Artículo en Inglés | MEDLINE | ID: mdl-33763734

RESUMEN

The novel corona (SARS-CoV-2) virus causes a global pandemic, which motivates researchers to develop reliable and effective methods for screening and detection of SARS-CoV-2. Though there are several methods available for the diagnosis of SARS-CoV-2 such as RT-PCR and ELSIA, nevertheless, these methods are time-consuming and may not apply at the point of care. In this study, we have developed a specific, sensitive, quantitative and fast detection method for SARS-CoV-2 by fluorescence resonance energy transfer (FRET) assay. The total extracellular protease proteolytic activity from the virus has been used as the biomarker. The specific peptide sequences from the library of 115 dipeptides were identified via changes in the fluorescence signal. The fluorogenic dipeptide substrates have the fluorophore and a quencher at the N- and the C- terminals, respectively. When the protease hydrolyzes the peptide bond between the two specific amino acids, it leads to a significant increase in the fluorescence signals. The specific fluorogenic peptide (H-d) produces a high fluorescence signal. A calibration plot was obtained from the changes in the fluorescence intensity against the different concentrations of the viral protease. The lowest limit of detection of this method was 9.7 ± 3 pfu/mL. The cross-reactivity of the SARS-CoV-2-specific peptide was tested against the MERS-CoV which does not affect the fluorescence signal. A significant change in the fluorescence signal with patient samples indicates that this FRET-based assay might be applied for the diagnosis of SARS-CoV-2 patients. Graphical abstract.


Asunto(s)
/métodos , /metabolismo , Colorantes Fluorescentes/metabolismo , Péptidos/metabolismo , Proteínas Virales/metabolismo , Animales , Bioensayo , Chlorocebus aethiops , Transferencia Resonante de Energía de Fluorescencia , Humanos , Biblioteca de Péptidos , /metabolismo , Células Vero , Ensayo de Placa Viral
12.
J Vis Exp ; (168)2021 02 10.
Artículo en Inglés | MEDLINE | ID: mdl-33645569

RESUMEN

Eye disorders affect millions of people worldwide, but the limited availability of human tissues hinders their study. Mouse models are powerful tools to understand the pathophysiology of ocular diseases because of their similarities with human anatomy and physiology. Alterations in the retinal pigment epithelium (RPE), including changes in morphology and function, are common features shared by many ocular disorders. However, successful isolation and culture of primary mouse RPE cells is very challenging. This paper is an updated audiovisual version of the protocol previously published by Fernandez-Godino et al. in 2016 to efficiently isolate and culture primary mouse RPE cells. This method is highly reproducible and results in robust cultures of highly polarized and pigmented RPE monolayers that can be maintained for several weeks on Transwells. This model opens new avenues for the study of the molecular and cellular mechanisms underlying eye diseases. Moreover, it provides a platform to test therapeutic approaches that can be used to treat important eye diseases with unmet medical needs, including inherited retinal disorders and macular degenerations.


Asunto(s)
Disección , Cultivo Primario de Células/métodos , Epitelio Pigmentado de la Retina/citología , Animales , Bioensayo , Diferenciación Celular , Polaridad Celular , Separación Celular , Impedancia Eléctrica , Células Epiteliales/citología , Humanos , Ratones Endogámicos C57BL , Fagocitosis , Factores de Tiempo
13.
J Vis Exp ; (168)2021 02 11.
Artículo en Inglés | MEDLINE | ID: mdl-33645577

RESUMEN

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate their food environment. The fruit fly, Drosophila melanogaster, is a genetically tractable model organism that is widely used to decipher the molecular, cellular, and neural underpinnings of food preference. To analyze fly food preference, a robust feeding method is needed. Described here is a two-choice feeding assay, which is rigorous, cost-saving, and fast. The assay is Petri-dish-based and involves the addition of two different foods supplemented with blue or red dye to the two halves of the dish. Then, ~70 prestarved, 2-4-day-old flies are placed in the dish and allowed to choose between blue and red foods in the dark for about 90 min. Examination of the abdomen of each fly is followed by the calculation of the preference index. In contrast to multiwell plates, each Petri dish takes only ~20 s to fill and saves time and effort. This feeding assay can be employed to quickly determine whether flies like or dislike a particular food.


Asunto(s)
Bioensayo/métodos , Drosophila melanogaster/fisiología , Preferencias Alimentarias , Animales , Colorantes , Conducta Alimentaria , Indicadores y Reactivos , Inanición
14.
Int J Mol Sci ; 22(5)2021 Feb 25.
Artículo en Inglés | MEDLINE | ID: mdl-33668965

RESUMEN

Bacterial wilt (BW) disease from Ralstonia solanacearum is a serious disease and causes severe yield losses in chili peppers worldwide. Resistant cultivar breeding is the most effective in controlling BW. Thus, a simple and reliable evaluation method is required to assess disease severity and to investigate the inheritance of resistance for further breeding programs. Here, we developed a reliable leaf-to-whole plant spread bioassay for evaluating BW disease and then, using this, determined the inheritance of resistance to R. solanacearum in peppers. Capsicum annuum 'MC4' displayed a completely resistant response with fewer disease symptoms, a low level of bacterial cell growth, and significant up-regulations of defense genes in infected leaves compared to those in susceptible 'Subicho'. We also observed the spreading of wilt symptoms from the leaves to the whole susceptible plant, which denotes the normal BW wilt symptoms, similar to the drenching method. Through this, we optimized the evaluation method of the resistance to BW. Additionally, we performed genetic analysis for resistance inheritance. The parents, F1 and 90 F2 progenies, were evaluated, and the two major complementary genes involved in the BW resistance trait were confirmed. These could provide an accurate evaluation to improve resistant pepper breeding efficiency against BW.


Asunto(s)
Bioensayo/métodos , Capsicum/microbiología , Resistencia a la Enfermedad/genética , Patrón de Herencia/genética , Fitomejoramiento , Enfermedades de las Plantas/microbiología , Hojas de la Planta/microbiología , Ralstonia solanacearum/fisiología , Capsicum/genética , Segregación Cromosómica/genética , Progresión de la Enfermedad , Fenotipo , Enfermedades de las Plantas/genética
15.
Ecotoxicol Environ Saf ; 214: 112092, 2021 May.
Artículo en Inglés | MEDLINE | ID: mdl-33690008

RESUMEN

Over the last two decades, effect-directed analysis (EDA) gained importance as a seminal screening tool for tracking biological effects of environmental organic micro-pollutants (MPs). As EDA using high-performance liquid chromatography and bioassays is costly and time consuming, recent implementations of this approach have combined high-performance thin-layer chromatography (HPTLC) with effect-based methods (EBMs) using cell-based bioassays, enabling the detection of estrogenic, androgenic, genotoxic, photosystem II (PSII)- inhibiting, and dioxin-like sample components on a HPTLC plate. In the present study, the developed methodologies were applied as a HPTLC-based bioassay battery, to investigate toxicant elimination efficiency of wastewater treatment plants (WWTPs), and to characterize the toxic potential of landfill leachates. Activity levels detected in untreated landfill leachates, expressed as reference compound equivalence (EQ) concentration, were up to 16.8 µg ß-naphthoflavone-EQ L-1 (indicating the degree of dioxin-like activity), 1.9 µg estradiol-EQ L-1 (estrogenicity) and 8.3 µg diuron-EQ L­1 (PSII-inhibition), dropping to maximal concentrations of 47 ng ß-naphthoflavone-EQ L-1, 0.7 µg estradiol-EQ L-1 and 53.1 ng diuron-EQ L-1 following treatment. Bisphenol A (BPA) is suggested to be the main contributor to estrogenic activity, with concentrations determined by the planar yeast estrogen screen corresponding well to results from chemical analysis. In the investigated WWTP samples, a decrease of estrogenic activity of 6-100% was observed following treatment for most of the active fractions, except of a 20% increase in one fraction (Rf = 0.568). In contrast, androgenicity with concentrations up to 640 ng dihydrotestosterone-EQ L-1 was completely removed by treatment. Interestingly, genotoxic activity increased over the WWTP processes, releasing genotoxic fractions into receiving waters. We propose this combined HPTLC and EBM battery to contribute to an efficient, cheap, fast and robust screening of environmental samples; such an assay panel would allow to gain an estimate of potential biological effects for prioritization prior to substance identification, and its routine application will support an inexpensive identification of the toxicity drivers as a first tier in an EDA strategy.


Asunto(s)
Bioensayo/métodos , Contaminantes Químicos del Agua/toxicidad , Purificación del Agua , Compuestos de Bencidrilo , Cromatografía en Capa Delgada/métodos , Monitoreo del Ambiente/métodos , Estrógenos/toxicidad , Fenoles , Dibenzodioxinas Policloradas/análisis , Aguas Residuales/análisis , beta-naftoflavona
16.
Molecules ; 26(4)2021 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-33672045

RESUMEN

Hydrochar is a carbon-based material that can be used as soil amendment. Since the physical-chemical properties of hydrochar are mainly assigned to process parameters, we aimed at evaluating the organic fraction of different hydrochars through 13C-NMR and off-line TMAH-GC/MS. Four hydrochars produced with sugarcane bagasse, vinasse and sulfuric or phosphoric acids were analyzed to elucidate the main molecular features. Germination and initial growth of maize seedlings were assessed using hydrochar water-soluble fraction to evaluate their potential use as growth promoters. The hydrochars prepared with phosphoric acid showed larger amounts of bioavailable lignin-derived structures. Although no differences were shown about the percentage of maize seeds germination, the hydrochar produced with phosphoric acid promoted a better seedling growth. For this sample, the greatest relative percentage of benzene derivatives and phenolic compounds were associated to hormone-like effects, responsible for stimulating shoot and root elongation. The reactions parameters proved to be determinant for the organic composition of hydrochar, exerting a strict influence on molecular features and plant growth response.


Asunto(s)
Espectroscopía de Resonancia Magnética con Carbono-13 , Carbón Orgánico/química , Carbón Orgánico/farmacología , Cromatografía de Gases y Espectrometría de Masas , Desarrollo de la Planta/efectos de los fármacos , Compuestos de Amonio Cuaternario/química , Agua/química , Bioensayo , Raíces de Plantas/anatomía & histología , Raíces de Plantas/efectos de los fármacos , Brotes de la Planta/anatomía & histología , Brotes de la Planta/efectos de los fármacos , Semillas/efectos de los fármacos , Zea mays/efectos de los fármacos , Zea mays/crecimiento & desarrollo
17.
J Insect Sci ; 21(2)2021 Mar 01.
Artículo en Inglés | MEDLINE | ID: mdl-33693806

RESUMEN

Background odors produced by plants in the environment can interfere with the response of insects to a point-releasing attractant, especially when their compositions overlap. In this study, a series of binary choice tests was conducted in a wind tunnel to investigate whether background odors emitted from cherry, blueberry, blackberry, or raspberry fruits would affect the level of Drosophila suzukii (Matsumura) attraction to its symbiotic yeast, Hanseniaspora uvarum (Niehaus) (Saccharomycetales: Saccharomycetaceae). Whether an increase in the intensity of background odors would affect the attractiveness of H. uvarum to D. suzukii was also investigated, either by increasing the number of cherry or raspberry fruit per cup or by increasing the number of fruit cups surrounding the cup baited with the yeast. In wind tunnel assays, background fruit odors interfering with D. suzukii attraction to the yeast varied among fruit types. Raspberry odor inhibited the attractiveness of H. uvarum to the fly the most, followed by blackberry odor, whereas cherry and blueberry odors had no significant impact on the attraction. An increase in the intensity of odors by adding more cherry or raspberry fruit per cup did not increase the impact of fruit odor on the attraction; however, adding more raspberry cups around H. uvarum linearly decreased its attractiveness, suggesting that background host fruit abundance and likely increase in host odor may influence D. suzukii attraction to yeast odor depending on host species.


Asunto(s)
Drosophila , Frutas/fisiología , Hanseniaspora , Odorantes , Animales , Bioensayo/métodos , Arándanos Azules (Planta)/fisiología , Drosophila/microbiología , Drosophila/fisiología , Prunus avium/fisiología , Rubus/fisiología , Saccharomycetales , Simbiosis
18.
Wei Sheng Yan Jiu ; 50(1): 111-115, 2021 Jan.
Artículo en Chino | MEDLINE | ID: mdl-33517971

RESUMEN

OBJECTIVE: To compare the result of serum folate determined by improved microbial assay and electrochemiluminescence method, and to look for the relationship between them, so as to provide basis for the assessment of nutrition status of folate in population. METHODS: A total of 258 serum samples were examined by improved microbial assay and electrochemiluminescence method. The correlation and consistence of the two method were analyzed. RESULTS: The result showed that the correlation coefficient of the two method was 0. 885, which indicated that the result of two method were highly correlated. Results of Bland-Altman method showed that 94. 5% of the values were within the consistency limit, and the Kappa value of Kappa test was 0. 665. The result of consistency analysis showed that there were some differences between the two methods, and the result of serum folate tested by improved microbial assay were higher than that of electrochemiluminescence method in general. CONCLUSION: The result of serum folate tested by electrochemiluminescence were highly correlated with the improved microbial assay, yet there are some differences in the consistency result between the two methods. Evaluating the nutrition status of folate by electrochemiluminescence may lead to a higher number of folate deficiency.


Asunto(s)
Deficiencia de Ácido Fólico , Bioensayo , Ácido Fólico , Humanos , Estado Nutricional
19.
Water Res ; 193: 116859, 2021 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-33540341

RESUMEN

Bioassays are increasingly being implemented for water quality monitoring as targeted chemical analyses are not always sufficient for the detection of all emerging chemicals or transformation products. However, the interpretation of bioassay results remains challenging, in particular because a positive response does not necessarily indicate that there may be an increased risk. For this purpose, effect-based trigger (EBT) values have been introduced as thresholds above which action needs to be undertaken to determine the cause of the response. The goals of this study were to (i) evaluate various approaches used to determine EBT values and (ii) based on the findings, derive human health EBT values for Chemical Activated LUciferase gene eXpression (CALUX) in vitro bioassays used for routine monitoring of water quality in the Netherlands. Finally, (iii) an uncertainty analysis was carried out to determine the protective power of the derived EBT values and the chance that potentially harmful substances might not be detected. EBT values that can be implemented in routine monitoring could be determined for four of eight selected bioassays. These EBT were compared to bioassay results from routine water quality monitoring carried out in the Netherlands. Furthermore, a framework for the calculation and evaluation of derived EBT values for routine application to monitor drinking water and its sources is proposed.


Asunto(s)
Agua Potable , Contaminantes Químicos del Agua , Bioensayo , Monitoreo del Ambiente , Humanos , Países Bajos , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad , Calidad del Agua
20.
Ecotoxicol Environ Saf ; 212: 112010, 2021 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-33550081

RESUMEN

Coastal areas are under continuous and increasing pressure from different human activities. A mixture of contaminants (e.g. hydrocarbons, pesticides, persistent organic pollutants (POPs), emerging contaminants, and others), originating mainly from populated, industrialised and agricultural areas, can reach the marine environment through different means such as wastewater discharge, soil runoffs, leaching from agriculture, and volatilisation/deposition. In this context, marine sediments have increasingly been considered repositories for a variety of pollutants that can accumulate and be stored for long periods, acting as a secondary source of contaminants during subsequent dredging operation or vessel manoeuvring. Chemical and ecotoxicological analyses of sediments are routinely conducted to evaluate the potential hazard/risk to the environment, either on bulk sediment or elutriate. In general, sediment elutriates are commonly prepared according to ASTM Guide even if alternative protocols are proposed by USACE for the various condition that they have to represent. The goal of the present study was to determine if the toxicological properties of ASTMprepared elutriates are comparable to those obtained from the USACE protocol. Sediment coming from 3 harbours (Olbia, Cagliari, and Toulon), as part of the "Se.D.Ri.Port" Interreg Project, were processed to obtain elutriates according to ASTM Guide and USACE Dredging Elutriate protocol and tested with the sea urchin Paracentrotus lividus embryo development test. Moreover, the significance of different stirring times of water/sediment mixture (1 h, 3 h, and 24 h) was tested with both the ASTM and USACE protocol. In addition to the biological analysis, for each sediment sample, heavy metals concentration, granulometry, and organic matter were determined. Even if for the ports of Toulon and Cagliari, the ASTM and USACE elutriates showed comparable results with P. lividus bioassay, for the port of Olbia the two protocols showed different criticalities. Preliminary results show that for the site Olbia elutriates prepared with the USACE protocol resulted in higher toxicity than elutriates obtained with ASTM (p < 0.001). In conclusion, differences in preparation protocols appear to be significant and can lead to different results in biological testing. To overcome this problem and to obtain more reliable evaluations of risk to the environment, standardisation and regulation must be the next goals in sediment management procedure.


Asunto(s)
Desarrollo Embrionario/efectos de los fármacos , Monitoreo del Ambiente/métodos , Sedimentos Geológicos/química , Metales Pesados/toxicidad , Paracentrotus/efectos de los fármacos , Agua de Mar/química , Contaminantes Químicos del Agua/toxicidad , Animales , Bioensayo , Ecotoxicología , Humanos , Italia , Larva/efectos de los fármacos , Larva/crecimiento & desarrollo , Mar Mediterráneo , Metales Pesados/análisis , Paracentrotus/embriología , Contaminantes Químicos del Agua/análisis
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