RESUMEN
Self-regenerating trigger waves can spread rapidly through the crowded cytoplasm without diminishing in amplitude or speed, providing consistent, reliable, long-range communication. The macromolecular concentration of the cytoplasm varies in response to physiological and environmental fluctuations, raising the question of how or if trigger waves can robustly operate in the face of such fluctuations. Using Xenopus extracts, we find that mitotic and apoptotic trigger wave speeds are remarkably invariant. We derive a model that accounts for this robustness and for the eventual slowing at extremely high and low cytoplasmic concentrations. The model implies that the positive and negative effects of cytoplasmic concentration (increased reactant concentration vs. increased viscosity) are nearly precisely balanced. Accordingly, artificially maintaining a constant cytoplasmic viscosity during dilution abrogates this robustness. The robustness in trigger wave speeds may contribute to the reliability of the extremely rapid embryonic cell cycle.
Asunto(s)
Citoplasma , Mitosis , Xenopus laevis , Animales , Citoplasma/metabolismo , Apoptosis , Viscosidad , Extractos Celulares/química , Modelos Biológicos , Xenopus , Ciclo CelularRESUMEN
Ongoing research on cyanotoxins, driven by the socioeconomic impact of harmful algal blooms, emphasizes the critical necessity of elucidating the toxicological profiles of algal cell extracts and pure toxins. This study comprehensively compares Raphidiopsis raciborskii dissolved extract (RDE) and cylindrospermopsin (CYN) based on Daphnia magna assays. Both RDE and CYN target vital organs and disrupt reproduction, development, and digestion, thereby causing acute and chronic toxicity. Disturbances in locomotion, reduced behavioral activity, and weakened swimming capability in D. magna have also been reported for both RDE and CYN, indicating the insufficiency of conventional toxicity evaluation parameters for distinguishing between the toxic effects of algal extracts and pure cyanotoxins. Additionally, chemical profiling revealed the presence of highly active tryptophan-, humic acid-, and fulvic acid-like fluorescence compounds in the RDE, along with the active constituents of CYN, within a 15-day period, demonstrating the chemical complexity and dynamics of the RDE. Transcriptomics was used to further elucidate the distinct molecular mechanisms of RDE and CYN. They act diversely in terms of cytotoxicity, involving oxidative stress and response, protein content, and energy metabolism, and demonstrate distinct modes of action in neurofunctions. In essence, this study underscores the distinct toxicity mechanisms of RDE and CYN and emphasizes the necessity for context- and objective-specific toxicity assessments, advocating nuanced approaches to evaluate the ecological and health implications of cyanotoxins, thereby contributing to the precision of environmental risk assessments.
Asunto(s)
Alcaloides , Toxinas Bacterianas , Toxinas de Cianobacterias , Cianobacterias , Daphnia , Animales , Toxinas Bacterianas/toxicidad , Daphnia/efectos de los fármacos , Alcaloides/toxicidad , Cianobacterias/química , Uracilo/análogos & derivados , Uracilo/toxicidad , Extractos Celulares/química , Extractos Celulares/farmacología , Floraciones de Algas NocivasRESUMEN
High plasma levels of homocysteine (Hcy) promote the progression of neurodegenerative diseases. However, the mechanism by which Hcy mediates neurotoxicity has not been elucidated. We observed that upon incubation with Hcy, the viability of a neuroblastoma cell line Neuro2a declined in a dose-dependent manner, and apoptosis was induced within 48 h. The median effective concentration (EC50) of Hcy was approximately 5 mM. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) nuclear translocation and acylation has been implicated in the regulation of apoptosis. We found that nuclear translocation and acetylation of GAPDH increased in the presence of 5 mM Hcy and that higher levels of acetyltransferase p300/CBP were detected in Neuro2a cells. These findings implicate the involvement of GAPDH in the mechanism whereby Hcy induces apoptosis in neurons. This study highlights a potentially important pathway in neurodegenerative disorders, and a novel target pathway for neuroprotective therapy.
Asunto(s)
Animales , Conejos , Apoptosis/efectos de los fármacos , Fármacos Neuroprotectores/farmacología , Gliceraldehído-3-Fosfato Deshidrogenasas/metabolismo , Homocisteína/farmacología , Acetilación , Acetiltransferasas/análisis , Factores de Tiempo , Recuento de Células , Extractos Celulares/química , Núcleo Celular/metabolismo , Supervivencia Celular/fisiología , Inducción Enzimática , Western Blotting , Técnica del Anticuerpo Fluorescente , Apoptosis/fisiología , Fármacos Neuroprotectores/administración & dosificación , Línea Celular Tumoral , Factores de Transcripción p300-CBP/metabolismo , Homocisteína/administración & dosificaciónRESUMEN
BACKGROUND: Unstable generation of free radicals in the body are responsible for many degenerative diseases. A bloom forming algae Euglena tuba growing abundantly in the aquatic habitats of Cachar district in the state of Assam in North-East India was analysed for its phytochemical contents, antioxidant activity as well as free radical scavenging potentials. RESULTS: Based on the ability of the extract in ABTSâ¢+ radical cation inhibition and Fe3+ reducing power, the obtained results revealed the prominent antioxidant activity of the algae, with high correlation coefficient of its TEAC values to the respective phenolic and flavonoid contents. The extract had shown its scavenging activity for different free radicals and 41.89 ± 0.41 µg/ml, 5.83 ± 0.07 µg/ml, 278.46 ± 15.02 µg/ml and 223.25 ± 4.19 µg/ml were determined as the IC50 values for hydroxyl, superoxide, nitric oxide and hypochlorous acid respectively, which are lower than that of the corresponding reference standards. The phytochemical analysis also revealed that the phenolics, flavonoids, alkaloids, tannins and carbohydrates are present in adequate amount in the extract which was confirmed by HPLC analysis. CONCLUSIONS: The results showed that 70% methanol extract of the algae possesses excellent antioxidant and free radical scavenging properties.
Asunto(s)
Animales , Masculino , Ratones , Extractos Celulares/química , Depuradores de Radicales Libres/metabolismo , Sustancias Reductoras/metabolismo , Euglena/química , Antioxidantes/metabolismo , Oxidación-Reducción , Fenoles/análisis , Ácido Ascórbico/análisis , Taninos/análisis , Flavonoides/análisis , Peroxidación de Lípido/efectos de los fármacos , Cromanos/metabolismo , Cromatografía Líquida de Alta Presión , Metanol , Alcaloides/análisis , Microalgas , Glucosa/análisis , IndiaRESUMEN
Human papillomavirus E7 (HPV E7) is a viral oncoprotein that plays an important role in cervical carcinogenesis through binding with retinoblastoma protein (Rb). Inactivation of Rb by E7 is necessary but not sufficient for cellular transformation, suggesting other protein-protein interactions are required for E7-mediated cellular transformation aside from the interaction with Rb. However, studies on the oncogenic function of HPV E7 have been limited by its poor immunoreactivity. In this report, we show that the fixation of purified recombinant HPV E7 on blotted nitrocellulose membrane with glutaldehyde markedly enhanced the immunoreactivity of HPV E7 protein. Using HeLa and Caski cell line which are infected with HPV 18 and HPV 16, respectively, we demonstrated that native HPV E7 proteins also could be detected by this method. These results therefore can provide the experimental conditions for detection of HPV E7 proteins with greater sensitivity and may help to analyze E7 functions.
Asunto(s)
Humanos , Extractos Celulares/química , Línea Celular , Inmunoquímica/métodos , Proteínas Oncogénicas Virales/análisis , Papillomaviridae/químicaRESUMEN
El n-dodecil sarcosinato de sodio produce la solubilización del espermatozoide humano con algo de contaminación acrosomal. El extracto está compuesto por una mezcla de productos con pesos moleculares y características inmunológicas similares a las del plasma seminal humano. El hecho de que la antigenicidad se mantenga luego de la solubilización, hace que ésta sea un comienzo útil en el aislamiento de moléculas que juegan un papel en la infertilidad por causas inmunológicas
Asunto(s)
Humanos , Masculino , Extractos Celulares/análisis , Membrana Celular/efectos de los fármacos , Infertilidad Masculina/diagnóstico , Sarcosina , Espermatozoides/ultraestructura , Extractos Celulares/química , Extractos Celulares/inmunología , Infertilidad Masculina/inmunología , Interacciones Espermatozoide-Óvulo , Interacciones Espermatozoide-Óvulo/inmunología , Semen/química , Semen/inmunología , Espermatozoides/efectos de los fármacos , Espermatozoides/inmunologíaRESUMEN
El n-dodecil sarcosinato de sodio produce la solubilización del espermatozoide humano con algo de contaminación acrosomal. El extracto está compuesto por una mezcla de productos con pesos moleculares y características inmunológicas similares a las del plasma seminal humano. El hecho de que la antigenicidad se mantenga luego de la solubilización, hace que ésta sea un comienzo útil en el aislamiento de moléculas que juegan un papel en la infertilidad por causas inmunológicas