RESUMO
Gene therapy offers a promising avenue for treating ischemic diseases, yet its clinical efficacy is hindered by the limitations of single gene therapy and the high oxidative stress microenvironment characteristic of such conditions. Lipid-polymer hybrid vectors represent a novel approach to enhance the effectiveness of gene therapy by harnessing the combined advantages of lipids and polymers. In this study, we engineered lipid-polymer hybrid nanocarriers with tailored structural modifications to create a versatile membrane fusion lipid-nuclear targeted polymer nanodelivery system (FLNPs) optimized for gene delivery. Our results demonstrate that FLNPs facilitate efficient cellular uptake and gene transfection via membrane fusion, lysosome avoidance, and nuclear targeting mechanisms. Upon encapsulating Hepatocyte Growth Factor plasmid (pHGF) and Catalase plasmid (pCAT), HGF/CAT-FLNPs were prepared, which significantly enhanced the resistance of C2C12 cells to H2O2-induced injury in vitro. In vivo studies further revealed that HGF/CAT-FLNPs effectively alleviated hindlimb ischemia-induced gangrene, restored motor function, and promoted blood perfusion recovery in mice. Metabolomics analysis indicated that FLNPs didn't induce metabolic disturbances during gene transfection. In conclusion, FLNPs represent a versatile platform for multi-dimensional assisted gene delivery, significantly improving the efficiency of gene delivery and holding promise for effective synergistic treatment of lower limb ischemia using pHGF and pCAT.
Assuntos
Terapia Genética , Isquemia , Lipídeos , Polímeros , Animais , Isquemia/terapia , Terapia Genética/métodos , Lipídeos/química , Camundongos , Polímeros/química , Nanopartículas/química , Fator de Crescimento de Hepatócito/genética , Linhagem Celular , Transfecção/métodos , Plasmídeos/genética , Técnicas de Transferência de Genes , Masculino , Membro Posterior/irrigação sanguínea , Catalase/metabolismoRESUMO
Biocatalytic degradation of non-hydrolyzable plastics is a rapidly growing field of research, driven by the global accumulation of waste. Enzymes capable of cleaving the carbon-carbon bonds in synthetic polymers are highly sought-after as they may provide tools for environmentally friendly plastic recycling. Despite some reports of oxidative enzymes acting on non-hydrolyzable plastics, including polyethylene or poly(vinyl chloride), the notion that these materials are susceptible to efficient enzymatic degradation remains controversial, partly driven by a general lack of studies independently reproducing previous observations. Here, we attempt to replicate two recent studies reporting that deconstruction of polyethylene and poly(vinyl chloride) can be achieved using an insect hexamerin from Galleria mellonella (so-called "Ceres") or a bacterial catalase-peroxidase from Klebsiella sp., respectively. Reproducing previously described experiments, we do not observe any activity on plastics using multiple reaction conditions and multiple substrate types. Digging deeper into the discrepancies between the previous data and our observations, we show how and why the original experimental results may have been misinterpreted.
Assuntos
Biodegradação Ambiental , Klebsiella , Polietileno , Cloreto de Polivinila , Polietileno/metabolismo , Polietileno/química , Animais , Cloreto de Polivinila/química , Cloreto de Polivinila/metabolismo , Klebsiella/enzimologia , Klebsiella/metabolismo , Catalase/metabolismo , Catalase/química , Proteínas de Insetos/metabolismo , Proteínas de Insetos/química , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/química , BiocatáliseRESUMO
In the in vitro motility assay (IVMA), actin filaments are observed while propelled by surface-adsorbed myosin motor fragments such as heavy meromyosin (HMM). In addition to fundamental studies, the IVMA is the basis for a range of lab-on-a-chip applications, e.g. transport of cargoes in nanofabricated channels in nanoseparation/biosensing or the solution of combinatorial mathematical problems in network-based biocomputation. In these applications, prolonged myosin function is critical as is the potential to repeatedly exchange experimental solutions without functional deterioration. We here elucidate key factors of importance in these regards. Our findings support a hypothesis that early deterioration in the IVMA is primarily due to oxygen entrance into in vitro motility assay flow cells. In the presence of a typically used oxygen scavenger mixture (glucose oxidase, glucose, and catalase), this leads to pH reduction by a glucose oxidase-catalyzed reaction between glucose and oxygen but also contributes to functional deterioration by other mechanisms. Our studies further demonstrate challenges associated with evaporation and loss of actin filaments with time. However, over 8 h at 21-26 °C, there is no significant surface desorption or denaturation of HMM if solutions are exchanged manually every 30 min. We arrive at an optimized protocol with repeated exchange of carefully degassed assay solution of 45 mM ionic strength, at 30 min intervals. This is sufficient to maintain the high-quality function in an IVMA over 8 h at 21-26 °C, provided that fresh actin filaments are re-supplied in connection with each assay solution exchange. Finally, we demonstrate adaptation to a microfluidic platform and identify challenges that remain to be solved for real lab-on-a-chip applications.
Assuntos
Actomiosina , Dispositivos Lab-On-A-Chip , Actomiosina/metabolismo , Actomiosina/química , Citoesqueleto de Actina/metabolismo , Oxigênio/metabolismo , Animais , Glucose Oxidase/metabolismo , Glucose Oxidase/química , Glucose/metabolismo , Concentração de Íons de Hidrogênio , Catalase/metabolismoRESUMO
Salt-processed Psoraleae Fructus is a commonly used tonic in clinical practice. However, its usage is restricted due to the inherent toxicity. The covalent modification of proteins by reactive metabolites(RMs) plays a role in the hepatotoxicity of salt-processed Psoraleae Fructus. This study delves into the protein covalent modification by RMs generated from psoralen/isopsoralen, the primary toxic components of salt-processed Psoraleae Fructus, by liquid chromatography-mass spectrometry(LC-MS), aiming to elucidate the mechanism underlying the hepatic injury induced by salt-processed Psoraleae Fructus. Biochemical methods were utilized to measure the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), catalase(CAT), malondialdehyde(MDA), superoxide dismutase(SOD), reduced glutathione(GSH), and glutathione S-transferase(GST) in mice. The pathological changes in the liver were observed by hematoxylin-eosin(HE) staining. Subsequently, ultra performance liquid chromatography-quadrupole-time-of-flight-mass spectrometry(UPLC-Q-TOF-MS) was employed to identify the primary toxic components of psoralen/isopsoralen and the RMs in salt-processed Psoraleae Fructus. Covalent bonding adducts of the toxic components/RMs with GSH and free amino acids were identified to investigate the effects of the toxic components on modification sites and patterns of amino acids. The modifications of RMs were incorporated into the variable modifications of Proteome Discoverer, and the target proteins of psoralen/isopsoralen were detected by liquid chromatography-quadrupole exactive-mass spectrometry. Lastly, Label-free quantitative proteomics was adopted to screen differential proteins, which were further subjected to KEGG and GO enrichment analyses and confirmed by qPCR. The results indicated that compared with the control group, salt-processed Psoraleae Fructus significantly elevated the ALT, AST, and MDA levels and lowered the SOD, CAT, GSH, and GST levels in a dose-dependent manner, while causing obvious vacuolization and inflammatory cell infiltration in mouse hepatocytes. Furthermore, the livers of mice in the salt-processed Psoraleae Fructus group showed the presence of five RMs of psoralen/isopsoralen, two adducts with GSH, and one adduct with cysteine. In addition, 10 proteins modified by the RMs of psoralen/isopsoralen were identified. A total of 133 differential proteins were detected in the livers of mice in the salt-processed Psoraleae Fructus group, including 92 with up-regulated expression and 41 with down-regulated expression. These differential proteins mainly involved ribosomes, rRNAs, and glutathione, affecting the proteasome pathway. The qPCR results were consistent with the differential proteins. These findings suggest that the RMs of psoralen/isopsoralen can covalently bind to GSH and modify cysteine and lysine residues of liver proteins. This covalent modification of proteins by harmful substances can potentially result in liver damage. Therefore, it can be inferred that the oxidative stress damage induced by salt-processed Psoraleae Fructus may be associated with the abnormality of proteasome and its complex, biosynthesis of ribosomes and their nucleoprotein complex, rRNA binding, and glutathione binding.
Assuntos
Doença Hepática Induzida por Substâncias e Drogas , Fígado , Psoralea , Animais , Camundongos , Psoralea/química , Fígado/efeitos dos fármacos , Fígado/metabolismo , Doença Hepática Induzida por Substâncias e Drogas/metabolismo , Masculino , Medicamentos de Ervas Chinesas/química , Superóxido Dismutase/metabolismo , Malondialdeído/metabolismo , Alanina Transaminase/metabolismo , Etanol/química , Etanol/toxicidade , Aspartato Aminotransferases/metabolismo , Aspartato Aminotransferases/genética , Humanos , Catalase/metabolismo , Proteínas/química , Proteínas/metabolismo , Frutas/química , Espectrometria de Massas , Glutationa/metabolismoRESUMO
BACKGROUND: Oxidative damage to the ovaries is the primary cause of impaired reproductive functions in female animals. This study aimed to investigate the protective role of N-Acetyl-L-cysteine (NAC) in reducing oxidative damage in the ovaries of female rabbits. METHODS AND RESULTS: Female rabbit ovaries were treated in vitro with varying concentrations of D-galactose (D-gal): 0, 5, 10, and 15 mg/mL, and it was found that 10 mg/mL D-gal significantly disrupted follicular structures, causing disarray in granulosa cell arrangements and significantly reducing T-SOD and GSH levels (p < 0.01). Consequently, we selected 10 mg/mL D-gal to establish an ovarian failure model. These models were treated with multiple doses of NAC (0, 0.1, 0.3, 0.5 mg/mL). The results revealed that the disruption in granulosa cell arrangement caused by 10 mg/mL D-gal was effectively alleviated by 0.1 mg/mL NAC compared to the D-gal treatment group. Furthermore, 10 mg/mL D-gal significantly (p < 0.01) reduced GSH, T-SOD, and catalase (CAT) levels in the ovaries. However, 0.1 mg/mL NAC effectively (p < 0.01) suppressed these adverse effects. Moreover, the current results showed that 10 mg/mL D-gal alone significantly (p < 0.01) downregulated the expression of Nrf2, GPX, PRDX4, GSR, SOD1, and TAF4B, whereas 0.1 mg/mL NAC counteracted these suppressive effects (p < 0.01). CONCLUSIONS: It could be concluded that NAC may delay ovarian failure by reducing D-gal-induced ovarian oxidative damage in female rabbit, suggested NAC could be a promising therapeutic agent for protecting against ovarian failure and potentially delaying ovarian failure in female rabbits.
Assuntos
Acetilcisteína , Galactose , Ovário , Estresse Oxidativo , Animais , Coelhos , Feminino , Acetilcisteína/farmacologia , Galactose/efeitos adversos , Galactose/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Ovário/efeitos dos fármacos , Ovário/metabolismo , Ovário/patologia , Insuficiência Ovariana Primária/induzido quimicamente , Insuficiência Ovariana Primária/metabolismo , Insuficiência Ovariana Primária/patologia , Células da Granulosa/metabolismo , Células da Granulosa/efeitos dos fármacos , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Superóxido Dismutase/metabolismo , Glutationa/metabolismo , Catalase/metabolismo , Modelos Animais de DoençasRESUMO
OBJECTIVES: Oxidative stress, an imbalance between the body's natural antioxidant defenses and the production of reactive oxygen species (ROS), can result in serious oral diseases, including oral cancer, periodontal diseases, and oral lichen planus, through the activation of the redox-sensitive transcription factors and inflammation. The purpose of this study was to assess the potential effects of a removable complete denture on the levels of oxidative stress markers, such as lipid peroxidation (MDA), advanced oxidation protein products (AOPP), and catalase, and the quantitative expression of the redox-sensitive transcription factor NF-κB p65 subunit. MATERIALS AND METHODS: This interventional follow-up study enrolled 40 participants of both sexes aged 28-78 years, with a median age of 56 years, where unstimulated saliva was collected before denture placement, immediately after the denture placement, and 24 h, 7 days, and 30 days after the denture placement. The most prominent ROS overproduction was reported on the seventh day (p < 0.05), followed by a significant fall in antioxidative defense. RESULTS: The NF-κB p65 subunit, whose expression pattern was highest in the same time period on the seventh day, serves as a signaling molecule for redox imbalance due to ROS production. Over the next 30 days, its levels remained moderately increased compared to the basal value, which may influence pro-inflammatory pathways and the integrity of oral tissue components. These alterations may be induced by the dentures, which can produce high pressures on the supporting tissues or by the synthetic materials used for producing the dentures. CONCLUSION: Our research may help to clarify the potential pathways by which oxidative stress and redox-sensitive inflammatory mediators, as well as mechanical and chemical irritants, may serve as risk factors for premalignant lesions in the mouth. Further research on this topic is required to understand the molecular mechanisms behind the relationship between inflammation and oral premalignant lesions caused by mechanical and chemical irritation.
Assuntos
Antioxidantes , Biomarcadores , Oxirredução , Estresse Oxidativo , Espécies Reativas de Oxigênio , Humanos , Pessoa de Meia-Idade , Masculino , Feminino , Idoso , Adulto , Biomarcadores/metabolismo , Biomarcadores/análise , Seguimentos , Espécies Reativas de Oxigênio/metabolismo , Antioxidantes/metabolismo , Saliva/química , Saliva/metabolismo , Inflamação/metabolismo , Prótese Total/efeitos adversos , NF-kappa B/metabolismo , Peroxidação de Lipídeos , Catalase/metabolismo , Produtos da Oxidação Avançada de Proteínas/metabolismo , Boca/metabolismo , Fator de Transcrição RelA/metabolismoRESUMO
Coastal ecosystems are characterized by various human activities with potential adverse impacts. This study aimed to evaluate the potential oxidative stress effects in representative aquatic biota deployed in situ at a sawmill wastes dump (test site) and reference site in a coastal ecosystem for a short term (28 days) period. PAHs and OCPs were analysed using GC-FID and GC-MS respectively in surface water and sediments. Oxidative stress indices (malondialdehyde, glutathione-s-transferase, reduced glutathione, catalase and superoxide dismutase) were evaluated following standard methods in Coptodon guineensis (Guinean Tilapia) and Callinectes amnicola (Blue crab) over a period of 28 days. Sum PAHs in the test site sediments, oxidative stress indices in C. guineensis liver and C. amnicola haemolymph after 28 days exposure were significantly higher (p < 0.0.5) compared to the reference site. The results showed the adverse impacts to biota of sawmill wastes which are continuously burnt at the test site with potential for long-term effects. Sustainable sawmill wastes management at the test site are recommended to sustain life below water (UNSDG 14).
Assuntos
Braquiúros , Monitoramento Ambiental , Estresse Oxidativo , Tilápia , Poluentes Químicos da Água , Animais , Poluentes Químicos da Água/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Ecossistema , Superóxido Dismutase/metabolismo , Catalase/metabolismoRESUMO
Plants produce reactive oxygen species (ROS) upon infection, which typically trigger defence mechanisms and impede pathogen proliferation. Root-knot nematodes (RKNs, Meloidogyne spp.) represent highly detrimental pathogens capable of parasitizing a broad spectrum of crops, resulting in substantial annual agricultural losses. The involvement of ROS in RKN parasitism is well acknowledged. In this study, we identified a novel effector from Meloidogyne incognita, named CATLe, that contains a conserved catalase domain, exhibiting potential functions in regulating host ROS levels. Phylogenetic analysis revealed that CATLe is conserved across RKNs. Temporal and spatial expression assays showed that the CATLe gene was specifically up-regulated at the early infection stages and accumulated in the subventral oesophageal gland cells of M. incognita. Immunolocalization demonstrated that CATLe was secreted into the giant cells of the host plant during M. incognita parasitism. Transient expression of CATLe significantly dampened the flg22-induced ROS production in Nicotiana benthamiana. In planta assays confirmed that M. incognita can exploit CATLe to manipulate host ROS levels by directly degrading H2O2. Additionally, interfering with expression of the CATLe gene through double-stranded RNA soaking and host-induced gene silencing significantly attenuated M. incognita parasitism, highlighting the important role of CATLe. Taken together, our results suggest that RKNs can directly degrade ROS products using a functional catalase, thereby manipulating host ROS levels and facilitating parasitism.
Assuntos
Catalase , Peróxido de Hidrogênio , Nicotiana , Espécies Reativas de Oxigênio , Tylenchoidea , Animais , Peróxido de Hidrogênio/metabolismo , Tylenchoidea/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Nicotiana/parasitologia , Catalase/metabolismo , Catalase/genética , Doenças das Plantas/parasitologia , Raízes de Plantas/parasitologia , Filogenia , Proteínas de Helminto/metabolismo , Proteínas de Helminto/genética , Interações Hospedeiro-ParasitaRESUMO
Background: In the realm of wheat seed germination, abiotic stresses such as salinity and high temperature have been shown to hinder the process. These stresses can lead to the production of reactive oxygen species, which, within a certain concentration range, may actually facilitate seed germination. γ-aminobutyric acid (GABA), a non-protein amino acid, serves as a crucial signaling molecule in the promotion of seed germination. Nevertheless, the potential of GABA to regulate seed germination under the simultaneous stress of heat and salinity remains unexplored in current literature. Methods: This study employed observational methods to assess seed germination rate (GR), physiological methods to measure H2O2 content, and the activities of glutamate decarboxylase (GAD), NADPH oxidase (NOX), superoxide dismutase (SOD), and catalase (CAT). The levels of ABA and GABA were quantified using high-performance liquid chromatography technology. Furthermore, quantitative real-time PCR technology was utilized to analyze the expression levels of two genes encoding antioxidant enzymes, MnSOD and CAT. Results: The findings indicated that combined stress (30 °C + 50 mM NaCl) decreased the GR of wheat seeds to about 21%, while treatment with 2 mM GABA increased the GR to about 48%. However, the stimulatory effect of GABA was mitigated by the presence of ABA, dimethylthiourea, and NOX inhibitor, but was strengthened by H2O2, antioxidant enzyme inhibitor, fluridone, and gibberellin. In comparison to the control group (20 °C + 0 mM NaCl), this combined stress led to elevated levels of ABA, reduced GAD and NOX activity, and a decrease in H2O2 and GABA content. Further investigation revealed that this combined stress significantly suppressed the activity of superoxide dismutase (SOD) and catalase (CAT), as well as downregulated the gene expression levels of MnSOD and CAT. However, the study demonstrates that exogenous GABA effectively reversed the inhibitory effects of combined stress on wheat seed germination. These findings suggest that GABA-induced NOX-mediated H2O2 signalling plays a crucial role in mitigating the adverse impact of combined stress on wheat seed germination. This research holds significant theoretical and practical implications for the regulation of crop seed germination by GABA under conditions of combined stress.
Assuntos
Germinação , Peróxido de Hidrogênio , Sementes , Triticum , Ácido gama-Aminobutírico , Peróxido de Hidrogênio/metabolismo , Triticum/efeitos dos fármacos , Triticum/metabolismo , Triticum/crescimento & desenvolvimento , Triticum/genética , Germinação/efeitos dos fármacos , Ácido gama-Aminobutírico/metabolismo , Sementes/efeitos dos fármacos , Sementes/crescimento & desenvolvimento , Transdução de Sinais/efeitos dos fármacos , Catalase/metabolismo , Catalase/genética , Estresse Salino/efeitos dos fármacos , Resposta ao Choque Térmico/efeitos dos fármacos , Resposta ao Choque Térmico/fisiologia , Superóxido Dismutase/metabolismo , Superóxido Dismutase/genéticaRESUMO
Di(2-ethylhexyl)phthalate (DEHP) is one of the most widely used plasticizers in plastic manufacturing. However, the toxicological effects of DEHP on dandelions remain poorly understood. This study comprehensively analyzed and explored the response mechanisms of dandelions to 1, 10, 50, and 100 mg L-1 DEHP influencing the morphophysiological growth, metabolomics, and molecular docking. DEHP reduced chlorophyll synthesis, inhibited plant growth, and induced oxidative-state-associated stress, which was manifested by the excessive production of reactive oxygen species, an increase in antioxidant enzyme activities, and enhanced synthesis of some osmoregulatory compounds, including proline and soluble protein. An analysis of the integrated biological response index showed that the toxicity was dose-dependent. Molecular docking demonstrated that DEHP could bind stably to three enzymes, and the binding energy was peroxidase (POD) > catalase (CAT) > superoxide dismutase (SOD). Metabolomics revealed that metabolite abundance and metabolic pathways were altered by DEHP, with 88 and 72 primary metabolites identified in shoots and roots, respectively. Amino acid, sugar, and organic acid metabolism were severely disturbed, with the most significant effects being on carbohydrate metabolism, valine, leucine, and isoleucine biosynthesis. Our study elucidated the influence of DEHP exposure on dandelions, providing new insights into the toxicity mechanisms and toxicological risk assessment.
Assuntos
Dietilexilftalato , Simulação de Acoplamento Molecular , Plastificantes , Taraxacum , Dietilexilftalato/toxicidade , Plastificantes/toxicidade , Taraxacum/efeitos dos fármacos , Superóxido Dismutase/metabolismo , Catalase/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Antioxidantes/metabolismo , Clorofila/metabolismo , Metabolômica , Peroxidase/metabolismoRESUMO
The toxicological hazard of iron-containing products is a public health concern that inspires research in identifying and developing readily available, inexpensive antidotes. Natural products, like plant-sourced antioxidants, can be of great value in this regard. Hesperetin a flavonoid abundantly present in citrus fruits is known to possess a diverse pharmacological and antioxidant attribute. The present study investigated the alleviation of detrimental effects of ferrous sulphate (FeSO4) by hesperetin in Drosophila melanogaster. Flies were exposed to FeSO4 (10 µM) alone or supplemented with hesperetin (50 or 100 µM) via diet for 7 consecutive days. Antioxidant enzyme activities, non-enzymatic antioxidant levels, acetylcholinesterase activity and oxidative stress markers were then measured. Hesperetin supplementation significantly (p < 0.05) attenuated FeSO4-induced oxidative stress by enhancement of enzymic antioxidants (catalase and glutathione-S-transferases) activities, preservation of non-enzymic antioxidants (total thiols and non-protein thiols), and reduction of other markers of oxidative stress (hydrogen peroxide, protein carbonyl and lipid peroxidation) in D. melanogaster. In addition, hesperetin supplementation decreased nitric oxide levels and enhanced acetylcholinesterase activity. Furthermore, hesperetin supplementation improved FeSO4-induced locomotor deficit, while there was no significant difference in cell viability (mitochondrial metabolic rate) in the treatment groups. This study suggests that hesperetin might be a promising functional agent in preventing iron toxicity and similar metal-induced impairments.
Assuntos
Acetilcolinesterase , Antioxidantes , Drosophila melanogaster , Compostos Ferrosos , Hesperidina , Estresse Oxidativo , Animais , Drosophila melanogaster/efeitos dos fármacos , Hesperidina/farmacologia , Compostos Ferrosos/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Acetilcolinesterase/metabolismo , Antioxidantes/farmacologia , Peroxidação de Lipídeos/efeitos dos fármacos , Masculino , Catalase/metabolismoRESUMO
The study investigated the impact of N-acetyl cysteine on growth, immune response, and antioxidant activity in tilapia (Oreochromis niloticus). Fish were reared at three densities (1.50, 3.00, and 4.50 kg/m3) with four levels of N-acetyl cysteine supplementation (0, 2, 4, and 6 mg/kg) over 60 days. Better growth was observed at low density, but at all densities, fish fed the highest N-acetyl cysteine level (6 mg/kg) showed improved growth. Chemical composition of fish and activity of amylase, lipase and protease in all treatments were noted to be insignificant. The levels of antioxidant enzymes (catalase, superoxide dismutase and glutathione peroxidase) and cortisol in HD treatments were high as compared to LD and MD treatment. However, fish fed with N3 diet in each density treatment showed the lowest level of antioxidant enzymes as well as cortisol. Similarly, the levels of malondialdehyde were noted to be high at HD treatments as compared to that in LD and MD. Its levels were lower in fish fed with N3 diets in all density treatments. Expression of somatostatins-1 did not increase in MD and HD treatments in response to high stocking density when compared with LD treatment. However, pro-opiomelanocortin-α level was reduced after N3 diet in HD treatment and interleukin 1-ß expression increased after N3 supplement in HD treatment. In conclusion, N-acetyl cysteine supplementation improved growth and antioxidant response in tilapia. The most optimum dose of N-acetyl cysteine was noted to be 6 mg/kg at high stocking, suggesting the potential role of this nutraceutical in tilapia intensive culture.
Assuntos
Acetilcisteína , Antioxidantes , Suplementos Nutricionais , Animais , Antioxidantes/metabolismo , Antioxidantes/farmacologia , Acetilcisteína/farmacologia , Hidrocortisona/metabolismo , Ciclídeos/crescimento & desenvolvimento , Ciclídeos/imunologia , Ciclídeos/metabolismo , Superóxido Dismutase/metabolismo , Ração Animal/análise , Catalase/metabolismo , Tilápia/crescimento & desenvolvimento , Tilápia/imunologia , Tilápia/metabolismo , Aquicultura/métodos , Glutationa Peroxidase/metabolismoRESUMO
Herbicides are the main class of pesticides applied in crops and are capable of polluting the surrounding freshwater system; thus, understanding their impact on non-target species, whose mechanism of action is not described, helps to elucidate the real risks of these pollutants to the environment. 2,4-dichlorophenoxyacetic acid (2,4-D) is frequently detected in water and, due to its persistence, poses a risk to wildlife. In this way, the present work aimed to describe the implication of exposure to concentrations of 2,4-D already reported in aquatic environments in several physiological mechanisms of C. riparius at molecular and biochemical levels. To achieve this, bioassays were conducted with fourth instar larvae exposed to three concentrations of 2,4-D (0.1, 1.0, and 7.5 µg L-1). Larvae were collected after 24 and 96 h of exposure, and the expression of 42 genes, related to six subcellular mechanisms, was assessed by Real-Time PCR (RT-PCR). Besides, the activity of the enzymes catalase (CAT), glutathione S-transferase (GST), and acetylcholinesterase (AChE) was determined. The main metabolic route altered after exposure to 2,4-D was the endocrine system (mainly related to 20-hydroxyecdysone and juvenile hormone), confirming its endocrine disruptor potential. Four of the eleven stress response genes studied were down-regulated, and later exposure modulated DNA-repair genes suggesting genotoxic capacity. Moreover, only one gene from each detoxification phase was modulated at short exposure to 1.0 µg L-1. The molecular responses were not dose-dependent, and some early responses were not preserved after 96 h, indicating a transient response to the herbicide. Exposure to 2,4-D did not alter the activity of CAT, GST, and AChE enzymes. The responses described in this study reveal new mechanistic pathways of toxicity for 2,4-D in non-target organisms and highlight potential ecological consequences for chironomids in aquatic systems at the edges of agricultural fields.
Assuntos
Ácido 2,4-Diclorofenoxiacético , Chironomidae , Glutationa Transferase , Herbicidas , Ácido 2,4-Diclorofenoxiacético/toxicidade , Animais , Chironomidae/efeitos dos fármacos , Chironomidae/genética , Herbicidas/toxicidade , Glutationa Transferase/metabolismo , Glutationa Transferase/genética , Acetilcolinesterase/metabolismo , Acetilcolinesterase/genética , Larva/efeitos dos fármacos , Larva/genética , Larva/metabolismo , Poluentes Químicos da Água/toxicidade , Catalase/metabolismo , Catalase/genética , Expressão Gênica/efeitos dos fármacosRESUMO
Plant essential oils (EOs)-based acaricides have been recognized as environmentally-friendly alternatives to synthetic acaricides because of their low toxicity against non-target species. Despite this, there are knowledge gaps regarding the toxicity mechanisms of plant EOs against non-target species. Here, the toxicology and enzymatic mechanism of Citrus reticulata and Citrus lemon EOs were evaluated against the vector pest, Haemaphysalis longicornis, and non-target ladybird beetle, Harmonia axyridis. Both EOs were mainly composed of d-Limonene, followed by ß-Myrcene and γ-Terpinene in C. reticulata, and (-)-ß-Pinene and γ-Terpinene in C. lemon. Citrus reticulata and C. lemon EOs were toxic to Hae. longicornis, with 50 % lethal concentration (LC50) values estimated at 0.43 and 0.98 µL/mL via nymphal immersion test, and 42.52 and 46.38 µL/mL via spray application, respectively. Among the constituents tested, ß-Myrcene was the most effective, with LC50 values of 0.17 and 47.87 µL/mL via immersion and spray treatment, respectively. A significant mortality of non-target Har. axyridis was found when treated by the EOs at concentrations two times greater than LC50 estimated against H. longicornis. The biochemical assay revealed that the EOs induced changes in the antioxidant enzyme activity of superoxide dismutases, catalase, and glutathione peroxidase in Hae. longicornis and Har. axyridis. The results demonstrated the acaricidal potential of citrus EOs and their major constituents for tick control, revealed the risk of the EOs to non-target species, and provided relevant insights into the mechanisms underlying their toxicity.
Assuntos
Acaricidas , Citrus , Besouros , Ixodidae , Óleos Voláteis , Animais , Óleos Voláteis/farmacologia , Óleos Voláteis/toxicidade , Besouros/efeitos dos fármacos , Ixodidae/efeitos dos fármacos , Ixodidae/enzimologia , Acaricidas/farmacologia , Acaricidas/toxicidade , Monoterpenos Cicloexânicos , Monoterpenos Bicíclicos/farmacologia , Monoterpenos Acíclicos/toxicidade , Monoterpenos Acíclicos/farmacologia , Limoneno/farmacologia , Monoterpenos/farmacologia , Monoterpenos/toxicidade , Cicloexenos/toxicidade , Cicloexenos/farmacologia , Terpenos/farmacologia , Catalase/metabolismo , Superóxido Dismutase/metabolismo , Glutationa Peroxidase/metabolismo , Antioxidantes/farmacologia , Haemaphysalis longicornisRESUMO
BACKGROUND: Tuberculosis (TB) is an airborne disease caused by Mycobacterium tuberculosis (M. tuberculosis). The world is currently facing challenges due to the spread of anti-tuberculosis drug-resistant of M. tuberculosis. Isoniazid-resistant (INH), is one of the first-line anti-tuberculosis agents that has a high resistance case. This study used Multiplex allele-specific Polymerase Chain Reaction (MAS-PCR) to detect the most common mutations associated with isoniazid resistance on inhA, katG, and ahpC gene. METHODS: This study used samples from clinical isolates of M. tuberculosis which had been tested for their antibiotic sensitivity of first-line anti-tuberculosis drugs. The DNA extraction process was carried out using the boiling method and then amplified with specific primers for inhA, katG, and ahpC genes using the MAS-PCR method. The results are then read on the electrophoretic gel with an interpretation of the mutation gene when the target gene DNA bands were absent according to the allele-specific fragments target. RESULTS: A total of 200 isolates were tested in this study consisting of isoniazid-resistant and susceptible with the largest distribution of Multi-Drug Resistant (MDR) isolates with a total of 146 isolates (73%). The most significant gene mutation was on the ahpC gene in 61 isolates (30,5%) and the combination mutation of the katG + ahpC gene in 52 isolates (26%) with sensitivity and specificity of the test reaching 87% and 42% for the detection of INH-resistant. CONCLUSION: Mutation on the ahpC gene has the highest percentage in this study. AhpC gene can be considered one of the essential genes to be tested for the cause of isoniazid-resistant. Using MAS-PCR for detecting gene mutation in isoniazid-resistant was simple and easy, it has the potential to be widely used as a rapid screening molecular test.
Assuntos
Antituberculosos , Proteínas de Bactérias , Catalase , Isoniazida , Mutação , Mycobacterium tuberculosis , Tuberculose Resistente a Múltiplos Medicamentos , Humanos , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/efeitos dos fármacos , Mycobacterium tuberculosis/isolamento & purificação , Indonésia , Isoniazida/farmacologia , Isoniazida/uso terapêutico , Antituberculosos/farmacologia , Antituberculosos/uso terapêutico , Proteínas de Bactérias/genética , Tuberculose Resistente a Múltiplos Medicamentos/microbiologia , Catalase/genética , Oxirredutases/genética , Testes de Sensibilidade Microbiana , Feminino , Masculino , Adulto , Reação em Cadeia da Polimerase Multiplex , Farmacorresistência Bacteriana Múltipla/genéticaRESUMO
Oxidative stress occurs during ex-situ heart perfusion (ESHP) and may negatively affect functional preservation of the heart. We sought to assess the status of key antioxidant enzymes during ESHP, and the effects of augmenting these antioxidants on the attenuation of oxidative stress and improvement of myocardial and endothelial preservation in ESHP. Porcine hearts were perfused for 6 hours with oxygen-derived free-radical scavengers polyethylene glycol (PEG)-catalase or PEG-superoxide dismutase (SOD) or with naive perfusate (control). The oxidative stress-related modifications were determined in the myocardium and coronary vasculature, and contractile function, injury, and endothelial integrity were compared between the groups. The activity of key antioxidant enzymes decreased and adding catalase and SOD restored the enzyme activity. Cardiac function and endothelial integrity were preserved better with restored catalase activity. Catalase and SOD both decreased myocardial injury and catalase reduced ROS production and oxidative modification of proteins in the myocardium and coronary vasculature. The activity of antioxidant enzymes decrease in ESHP. Catalase may improve the preservation of cardiac function and endothelial integrity during ESHP. While catalase and SOD may both exert cardioprotective effects, unbalanced SOD and catalase activity may paradoxically increase the production of reactive species during ESHP.
Assuntos
Catalase , Sequestradores de Radicais Livres , Estresse Oxidativo , Superóxido Dismutase , Animais , Suínos , Superóxido Dismutase/metabolismo , Catalase/metabolismo , Sequestradores de Radicais Livres/farmacologia , Estresse Oxidativo/efeitos dos fármacos , Perfusão/métodos , Miocárdio/metabolismo , Polietilenoglicóis/farmacologia , Coração/fisiologia , Coração/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Antioxidantes/farmacologia , Preservação de Órgãos/métodosRESUMO
Cotton aphids (Aphis gossypii Glover) cause harm by feeding on phloem sap and spreading plant viruses to lily. Understanding the mechanisms by which aphids infest lily plants is crucial for effective aphid management and control. In this study, we investigated the activity of antioxidants, integrated nontargeted metabolomes and transcriptomes of lilies infested by cotton aphids to explore the changes in lily leaves. Overall, the results indicated that the catalase (CAT) activity in the leaves of the lily plants was greater than that in the leaves of the control plants. A comprehensive identification of 604 substances was conducted in the leaves. Furthermore, the differentially abundant metabolite analysis revealed the enrichment of phenylalanine metabolism and α-linolenic acid metabolism. Moreover, 3574 differentially expressed genes (DEGs), whose expression tended to increase, were linked to glutathione metabolism and phenylpropanoid biosynthesis. In addition, the integrated analysis revealed that the defensive response of lily leaves to aphids is manifested through antioxidant reactions, phenylpropane and flavonoid biosynthesis, and α-linolenic acid metabolism. Finally, the key metabolites were CAT, glutathione, coumaric acid, and jasmonic acid, along with the key genes chalcone synthase (CHS), phenylalanine ammonia-lyase (PAL), and 12-oxo-phytodienoic acid reductase (OPR). Accordingly, the findings of this research elucidate the molecular and metabolic reactions of A. gossypii in lily plants, offering valuable insights for developing aphid resistance strategies in lily farming.
Assuntos
Antioxidantes , Afídeos , Lilium , Folhas de Planta , Transcriptoma , Afídeos/genética , Lilium/genética , Lilium/metabolismo , Animais , Folhas de Planta/genética , Folhas de Planta/metabolismo , Folhas de Planta/parasitologia , Antioxidantes/metabolismo , Regulação da Expressão Gênica de Plantas , Metabolômica/métodos , Metaboloma , Catalase/metabolismo , Catalase/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Extremely low-frequency electromagnetic fields (ELF-EMFs) are ubiquitous in industrialized environments due to the continuous use of electrical devices. Our previous studies demonstrated that ELF-EMFs affect muscle cells by modulating oxidative stress and enhancing myogenesis. This pilot study investigated these effects on the skeletal muscles of sedentary adult mice, assessing physiological responses to ELF-EMF exposure and potential modulation by antioxidant supplementation. Male C57BL/6 mice were exposed to ELF-EMFs (0.1 or 1.0 mT) for 1 h/day for up to 5 weeks and fed a standard diet without or with N-acetyl-cysteine (NAC). The results showed transient increases in muscle strength (after 2 weeks of exposure at 1.0 mT), potentially linked to muscle fiber recruitment and activation, revealed by higher PAX7 and myosin heavy chain (MyH) expression levels. After ELF-EMF exposure, oxidative status assessment revealed transient increases in the expression levels of SOD1 and catalase enzymes, in total antioxidant capacity, and in protein carbonyl levels, markers of oxidative damage. These effects were partially reduced by NAC. In conclusion, ELF-EMF exposure affects skeletal muscle physiology and NAC supplementation partially mitigates these effects, highlighting the complex interactions between ELF-EMFs and antioxidant pathways in vivo. Further investigations on ELF-EMFs as a therapeutic modality for muscle health are necessary.
Assuntos
Campos Eletromagnéticos , Camundongos Endogâmicos C57BL , Músculo Esquelético , Estresse Oxidativo , Superóxido Dismutase-1 , Animais , Campos Eletromagnéticos/efeitos adversos , Camundongos , Masculino , Projetos Piloto , Músculo Esquelético/metabolismo , Músculo Esquelético/efeitos da radiação , Estresse Oxidativo/efeitos da radiação , Superóxido Dismutase-1/metabolismo , Acetilcisteína/farmacologia , Cadeias Pesadas de Miosina/metabolismo , Antioxidantes/metabolismo , Fator de Transcrição PAX7/metabolismo , Comportamento Sedentário , Força Muscular/efeitos da radiação , Catalase/metabolismoRESUMO
PURPOSE: The main factor that causes cataracts is the increased oxidative stress and imbalance of an antioxidant defense mechanism, which leads to significant changes in the lens microarchitecture. Senile cataract is the most common type of acquired cataracts due to aging. METHODS: We carried out a case-control study in the biochemistry department to examine the antioxidant status (catalase and total antioxidant capacity [TAC]) and lipid peroxidation marker, that is, malondialdehyde (MDA) in human lens epithelial cells (HLECs) of different grades of senile cortical, nuclear, and posterior subcapsular cataracts. We collected 150 samples from patients aged 50-90 years. These included 50 samples of cortical cataracts, 50 of nuclear cataracts, and 50 samples of posterior subcapsular cataracts. We measured catalase activity by the Beer method, TAC by the Benzie and Strain method, and protein by the Bradford method. We also estimated TAC in the aqueous extract of HLECs by the ferric reducing ability of plasma (FRAP) method and MDA by the thiobarbituric acid assay method. RESULTS: The results of this study showed that the level of catalase enzyme was higher in the first grade of nuclear, posterior subcapsular, and cortical cataracts than in other grades. This suggests that the catalase enzyme activity drops sharply in the second and third grades of these types of cataracts. The same pattern was observed for TAC, which was higher in the first grade of nuclear, posterior subcapsular, and cortical cataracts than in other grades. There were significant differences between catalase and TAC in different grades of cataracts, indicating that as the grading increases, both catalase and TAC decrease. CONCLUSION: The results of this study showed that the levels of MDA were higher and the levels of catalase and TAC were lower in patients with more severe cataracts compared to the healthy controls.
Assuntos
Antioxidantes , Catalase , Catarata , Células Epiteliais , Cristalino , Estresse Oxidativo , Humanos , Catarata/metabolismo , Idoso , Feminino , Pessoa de Meia-Idade , Masculino , Células Epiteliais/metabolismo , Antioxidantes/metabolismo , Cristalino/metabolismo , Idoso de 80 Anos ou mais , Estudos de Casos e Controles , Catalase/metabolismo , Peroxidação de Lipídeos , Biomarcadores/metabolismo , Malondialdeído/metabolismoRESUMO
While vitiligo is primarily caused by melanocyte deficiency or dysfunction, recent studies have revealed a notable prevalence of metabolic syndrome (MetS) among patients with vitiligo. This suggests shared pathogenic features between the two conditions. Individuals with vitiligo often exhibit variations in triglyceride levels, cholesterol, and blood pressure, which are also affected in MetS. Given the similarities in their underlying mechanisms, genetic factors, pro-inflammatory signalling pathways, and increased oxidative stress, this study aims to highlight the common traits between vitiligo and metabolic systemic disorders. Serum analyses confirmed increased low-density lipoprotein (LDL) levels in patients with vitiligo, compared to physiological values. In addition, we reported significant decreases in folate and vitamin D (Vit D) levels. Oxidative stress is one of the underlying causes of the development of metabolic syndromes and is related to the advancement of skin diseases. This study found high levels of inflammatory cytokines, such as interleukin-6 (IL-6) and chemokine 10 (CXCL10), which are markers of inflammation and disease progression. The accumulation of insulin growth factor binding proteins 5 (IGFBP5) and advanced glycation end products (AGEs) entailed in atherosclerosis and diabetes onset, respectively, were also disclosed in vitiligo. In addition, the blood-associated activity of the antioxidant enzymes catalase (Cat) and superoxide dismutase (SOD) was impaired. Moreover, the plasma fatty acid (FAs) profile analysis showed an alteration in composition and specific estimated activities of FAs biosynthetic enzymes resembling MetS development, resulting in an imbalance towards pro-inflammatory n6-series FAs. These results revealed a systemic metabolic alteration in vitiligo patients that could be considered a new target for developing a more effective therapeutic approach.