Microcystin-LR acute exposure increases AChE activity via transcriptional ache activation in zebrafish (Danio rerio) brain

AbstractMicrocystins (MCs) constitute a family of cyanobacterial toxins, with more than 80 variants. These toxins are able to induce hepatotoxicity in several organisms mainly through the inhibition of protein phosphatases PP1 and PP2A and oxidative stress generation. Since recent evidence shows that MCs can either accumulate in brain or alter behavior patterns of fish species, in this study we tested the in vitro and in vivo effects of MC-LR at different concentrations on acetylcholinesterase (AChE) activity in zebrafish brain. In vivo studies showed that 100μg/L MC-LR led to a significant increase in the AChE activity (27%) when zebrafish were exposed to the toxin dissolved in water, but did not cause any significant changes when injected intraperitoneally. In addition, semiquantitative RT-PCR analysis demonstrated that 100μg/L MC-LR exposure also increased ache mRNA levels in zebrafish brain. The in vitro assays did not reveal any significant changes in AChE activity. These findings provide the first evidence that brain AChE is another potential target for MCs and suggest that the observed increases in AChE enzymatic activity and in ache transcript levels after MC-LR exposure depend, at least partially, on branchial uptake or ingestion

Intraperitoneal exposure to nano/microparticles of fullerene (C60) increases acetylcholinesterase activity and lipid peroxidation in adult zebrafish (danio rerio) brain

Even though technologies involving nano/microparticles have great potential, it is crucial to determine possible toxicity of these technological products before extensive use. Fullerenes C60 are nanomaterials with unique physicochemical and biological properties that are important for the development of many technological applications. The aim of this study was to evaluate the consequences of nonphotoexcited fullerene C60 exposure in brain acetylcholinesterase expression and activity, antioxidant responses, and oxidative damage using adult zebrafish as an animal model. None of the doses tested (7.5, 15, and 30 mg/kg) altered AChE activity, antioxidant responses, and oxidative damage when zebrafish were exposed to nonphotoexcited C60 nano/microparticles during 6 and 12 hours. However, adult zebrafish exposed to the 30 mg/kg dose for 24 hours have shown enhanced AChE activity and augmented lipid peroxidation (TBARS assays) in brain. In addition, the up-regulation of brain AChE activity was neither related to the transcriptional control (RT-qPCR analysis) nor to the direct action of nonphotoexcited C60 nano/microparticles on the protein (in vitro results) but probably involved a posttranscriptional or posttranslational modulation of this enzymatic activity. Taken together these findings provided further evidence of toxic effects on brain after C60 exposure

Avalia??o da atividade das ectonucleotidases em resposta aos efeitos adversos causados pela exposi??o a 2,3,7,8- Tetraclorodibenzeno-p-dioxina (TCDD) utilizando zebrafish (Danio rerio) como modelo de estudo

Made available in DSpace on 2015-04-14T14:51:13Z (GMT). No. of bitstreams: 1 434409.pdf: 489532 bytes, checksum: 6d9ad85140ac1052e70482873df7cc50 (MD5) Previous issue date: 2011-08-03A 2,3,7,8-tetraclorodibenzeno-p-dioxina (TCDD) ? membro de um grupo de contaminantes ambientais conhecido como hidrocarboneto arom?tico policlorinato e pode ser formada durante o processo de branqueamento com cloro, utilizado por f?brica de celulose e papel e como um subproduto da fabrica??o de determinados produtos qu?micos. Uma vez associado a ligantes como a TCDD, o receptor de hidrocarboneto arila (AHR) ? translocado para o n?cleo onde ir? formar um d?mero com o translocador nuclear do receptor hidrocarboneto arila (ARNT). O heterod?mero AHR/ARNT formado regula a express?o de uma s?rie de genes envolvidos na resposta celular a mudan?as ambientais e a condi??es de desenvolvimento. Entre os genes, est? o que codifica para NTPDase2 (previamente conhecida como ecto-ATPase). Com isso, torna-se importante avaliar a exposi??o aguda (in vivo) da TCDD sobre a hidr?lise de ectonucleotidases em membranas cerebrais de zebrafish. As ectonucleotidases s?o enzimas capazes de hidrolisar os nucleot?deos e inativar a sinaliza??o mediada pelos nucleot?deos extracelulares. Entre as ectonucleotidases, destacam-se as NTPDases e a ecto-5 -nucleotidase. A atividade de NTPDase demonstrou que n?o houve diferen?a significativa na hidr?lise de ATP e de ADP, bem como de AMP em membranas cerebrais de zebrafish. Estas descobertas parecem ser importantes para refor?ar a id?ia de que a indu??o do gene que codifica ? NTPDase2 pode n?o ser um fator geral da toxicidade de TCDD. No entanto, mais estudos utilizando doses mais elevadas de TCDD e exposi??o a longo prazo s?o necess?rias

Histological and Molecular Evidence of the Positive Performance of Glycerol-Plasticized Chitosan-Alginate Membranes on Skin Lesions of Hyperglycemic Mice

The purpose of this study was to investigate tissue repair of excisional wounds in hyperglycemic animals treated with chitosan-alginate membranes (CAM) produced in the presence of glycerol. 8-week C57B1 male mice were divided into normoglycemic animals with a 0.9% saline solution topical treatment (CTSF); hyperglycemic animals with 0.9% saline solution topical treatment (DMSF) and hyperglycemic animals with glycerol-plasticized chitosan-alginate membrane topical treatment (DMCAM). On post-wound day three, the DMCAM group presented a lower number of leukocytes, mature mastocytes, a higher number of vessels (p < 0.05), and active mastocytes (p < 0.05) when compared to the CTSF and DMSF groups. There were no differences regarding the distribution, deposition, organization, and thickness of collagen fibers. On day 7 there were no differences in the analysis of fibroblasts, mastocytes, and TGF−β1 and VEGF expressions among the groups. Regarding collagen fibers, the DMCAM group presented slight red-orange birefringence when compared to the CTSF and DMSF groups. On day 14 there was a slight concentration of thinner elastic fibers for the DMCAM group, with a greater reorganization of papillary skin and improved red-orange birefringence collagen fibers, as well as net-shaped orientation, similar to intact skin. In addition, improved elastic fiber organization distributed in the entire neo-dermis and a larger presence of elaunin fibers were observed, in a similar pattern found in the intact skin. The use of CAM in cutaneous lesions boosted tissue repair since there was a smaller number of inflammatory cells and mastocytes, and an improvement in collagen deposition and collagen fibers. These results demonstrate the high potential of plasticized chitosan-alginate membrane for skin wound dressing of hyperglycemic patients

Weaning from mechanical ventilation in intensive care units across 50 countries (WEAN SAFE): a multicentre, prospective, observational cohort study

Background Current management practices and outcomes in weaning from invasive mechanical ventilation are poorly understood. We aimed to describe the epidemiology, management, timings, risk for failure, and outcomes of weaning in patients requiring at least 2 days of invasive mechanical ventilation. Methods WEAN SAFE was an international, multicentre, prospective, observational cohort study done in 481 intensive care units in 50 countries. Eligible participants were older than 16 years, admitted to a participating intensive care unit, and receiving mechanical ventilation for 2 calendar days or longer. We defined weaning initiation as the first attempt to separate a patient from the ventilator, successful weaning as no reintubation or death within 7 days of extubation, and weaning eligibility criteria based on positive end-expiratory pressure, fractional concentration of oxygen in inspired air, and vasopressors. The primary outcome was the proportion of patients successfully weaned at 90 days. Key secondary outcomes included weaning duration, timing of weaning events, factors associated with weaning delay and weaning failure, and hospital outcomes. This study is registered with ClinicalTrials.gov, NCT03255109. Findings Between Oct 4, 2017, and June 25, 2018, 10 232 patients were screened for eligibility, of whom 5869 were enrolled. 4523 (77·1%) patients underwent at least one separation attempt and 3817 (65·0%) patients were successfully weaned from ventilation at day 90. 237 (4·0%) patients were transferred before any separation attempt, 153 (2·6%) were transferred after at least one separation attempt and not successfully weaned, and 1662 (28·3%) died while invasively ventilated. The median time from fulfilling weaning eligibility criteria to first separation attempt was 1 day (IQR 0–4), and 1013 (22·4%) patients had a delay in initiating first separation of 5 or more days. Of the 4523 (77·1%) patients with separation attempts, 2927 (64·7%) had a short wean (≤1 day), 457 (10·1%) had intermediate weaning (2–6 days), 433 (9·6%) required prolonged weaning (≥7 days), and 706 (15·6%) had weaning failure. Higher sedation scores were independently associated with delayed initiation of weaning. Delayed initiation of weaning and higher sedation scores were independently associated with weaning failure. 1742 (31·8%) of 5479 patients died in the intensive care unit and 2095 (38·3%) of 5465 patients died in hospital. Interpretation In critically ill patients receiving at least 2 days of invasive mechanical ventilation, only 65% were weaned at 90 days. A better understanding of factors that delay the weaning process, such as delays in weaning initiation or excessive sedation levels, might improve weaning success rates

Weaning from mechanical ventilation in intensive care units across 50 countries (WEAN SAFE): a multicentre, prospective, observational cohort study

Background: Current management practices and outcomes in weaning from invasive mechanical ventilation are poorly understood. We aimed to describe the epidemiology, management, timings, risk for failure, and outcomes of weaning in patients requiring at least 2 days of invasive mechanical ventilation. Methods: WEAN SAFE was an international, multicentre, prospective, observational cohort study done in 481 intensive care units in 50 countries. Eligible participants were older than 16 years, admitted to a participating intensive care unit, and receiving mechanical ventilation for 2 calendar days or longer. We defined weaning initiation as the first attempt to separate a patient from the ventilator, successful weaning as no reintubation or death within 7 days of extubation, and weaning eligibility criteria based on positive end-expiratory pressure, fractional concentration of oxygen in inspired air, and vasopressors. The primary outcome was the proportion of patients successfully weaned at 90 days. Key secondary outcomes included weaning duration, timing of weaning events, factors associated with weaning delay and weaning failure, and hospital outcomes. This study is registered with ClinicalTrials.gov, NCT03255109. Findings: Between Oct 4, 2017, and June 25, 2018, 10 232 patients were screened for eligibility, of whom 5869 were enrolled. 4523 (77·1%) patients underwent at least one separation attempt and 3817 (65·0%) patients were successfully weaned from ventilation at day 90. 237 (4·0%) patients were transferred before any separation attempt, 153 (2·6%) were transferred after at least one separation attempt and not successfully weaned, and 1662 (28·3%) died while invasively ventilated. The median time from fulfilling weaning eligibility criteria to first separation attempt was 1 day (IQR 0-4), and 1013 (22·4%) patients had a delay in initiating first separation of 5 or more days. Of the 4523 (77·1%) patients with separation attempts, 2927 (64·7%) had a short wean (≤1 day), 457 (10·1%) had intermediate weaning (2-6 days), 433 (9·6%) required prolonged weaning (≥7 days), and 706 (15·6%) had weaning failure. Higher sedation scores were independently associated with delayed initiation of weaning. Delayed initiation of weaning and higher sedation scores were independently associated with weaning failure. 1742 (31·8%) of 5479 patients died in the intensive care unit and 2095 (38·3%) of 5465 patients died in hospital. Interpretation: In critically ill patients receiving at least 2 days of invasive mechanical ventilation, only 65% were weaned at 90 days. A better understanding of factors that delay the weaning process, such as delays in weaning initiation or excessive sedation levels, might improve weaning success rates. Funding: European Society of Intensive Care Medicine, European Respiratory Society