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BACKGROUND: Immunotherapy with clodronate-encapsulated liposomes, which induce macrophage depletion, has been studied extensively. However, previously reported liposomal formulation-based drugs (Clodrosome® and m-Clodrosome®) are limited by their inconsistent size and therapeutic efficacy. Thus, we aimed to achieve consistent therapeutic effects by effectively depleting macrophages with uniform-sized liposomes. RESULTS: We developed four types of click chemistry-based liposome nanoplatforms that were uniformly sized and encapsulated with clodronate, for effective macrophage depletion, followed by conjugation with Man-N3 and radiolabeling. Functionalization with Man-N3 improves the specific targeting of M2 macrophages, and radioisotope labeling enables in vivo imaging of the liposome nanoplatforms. The functionalized liposome nanoplatforms are stable under physiological conditions. The difference in the biodistribution of the four liposome nanoplatforms in vivo were recorded using positron emission tomography imaging. Among the four platforms, the clodronate-encapsulated mannosylated liposome effectively depleted M2 macrophages in the normal liver and tumor microenvironment ex vivo compared to that by Clodrosome® and m-Clodrosome®. CONCLUSION: The newly-developed liposome nanoplatform, with finely tuned size control, high in vivo stability, and excellent ex vivo M2 macrophage targeting and depletion effects, is a promising macrophage-depleting agent.
Assuntos
Ácido Clodrônico , Lipossomos , Masculino , Humanos , Lipossomos/farmacologia , Ácido Clodrônico/farmacologia , Distribuição Tecidual , MacrófagosRESUMO
n-Butyl acrylate (nBA) is one of acrylate esters which has been applied to diverse industrial fields. For unveiling of xeno-estrogenic effects and oxidative stress induction by nBA under two-generational exposure regimen (17 weeks), the biomarkers relevant to an estrogenic effect and oxidative stress were analyzed. Acute toxicity value of nBA in Oryzias latipes was 7.2 mg/L (96 h-LC50). Over exposure time, the significant transcriptional change of cytochrome P450 19A (CYP19A) and vitellogenin 1/2 (VTG1/2) was not observed (one-way ANOVA, P < 0.05), meaning no estrogenic effect of nBA. Significant reduction of glutathione (GSH) content was observed in F0 male and female fish, while in F1 male, the content was increased (P < 0.05). Catalase (CAT) activity of male fish showed the significant decrease in both F0 and F1 fish, showing multi-generational suppressing effect of nBA on CAT activity. But in case of reactive oxygen species (ROS), expression level and glutathione S-transferase (GST) activity were not modulated in response to nBA. These findings suggest that nBA could affect an antioxidant system alteration through GSH depletion and inhibition of CAT activity which could be transferred to the next generation, whereas xeno-estrogenic effect would be questionable.
Assuntos
Acrilatos/toxicidade , Antioxidantes/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Oryzias/genética , Acrilatos/metabolismo , Adaptação Fisiológica/fisiologia , Animais , Feminino , Proteínas de Peixes/genética , Proteínas de Peixes/metabolismo , Fígado/metabolismo , Masculino , Oryzias/metabolismo , Testes de Toxicidade AgudaRESUMO
Pancreatic cancer is one of the most lethal cancers and remains a major unsolved health problem. Less than 20 % of patients are surgical candidates, and the median survival for non-resected patients is approximately 3 to 4 months. Despite the existence of many conventional cancer therapies, few targeted therapies have been developed for pancreatic cancer. Combination therapy using erlotinib and gemcitabine is an approved standard chemotherapy for advanced pancreatic cancer, but it has marginal therapeutic benefit. To try to improve the therapeutic outlook, we studied the efficacy of another combination treatment and the relevance to E-cadherin in human pancreatic cancer cells. We treated two human pancreatic cancer cell lines with the histone deacetylase inhibitor (HDACi) SAHA. Interestingly, in these Panc-1 and Capan1 cells, we observed that the expression levels of E-cadherin and phosphorylated EGFR were gradually upregulated after treatment with SAHA. Furthermore, these cells underwent induced cell death after exposure to the combination treatment of SAHA and erlotinib. In Panc-1 cells, overexpression of E-cadherin activated the phosphorylation of EGFR and increased the cell sensitivity to erlotinib. In Capan1 cells, knocking down E-cadherin decreased the expression of phosphorylated EGFR, and these cells did not respond to erlotinib. Therefore, we demonstrated the efficacy of the combined treatment with SAHA and erlotinib in human pancreatic cancer cells, and we determined that the increased efficacy was due, at least in part, to the effects of SAHA on the expression of E-cadherin. Our studies suggest that E-cadherin may be a potent biomarker for pancreatic cancer.
Assuntos
Caderinas/genética , Receptores ErbB/biossíntese , Cloridrato de Erlotinib/administração & dosagem , Inibidores de Histona Desacetilases/administração & dosagem , Ácidos Hidroxâmicos/administração & dosagem , Neoplasias Pancreáticas/tratamento farmacológico , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Caderinas/biossíntese , Linhagem Celular Tumoral , Desoxicitidina/administração & dosagem , Desoxicitidina/análogos & derivados , Resistencia a Medicamentos Antineoplásicos/genética , Receptores ErbB/genética , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Técnicas de Silenciamento de Genes , Humanos , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/patologia , Vorinostat , GencitabinaRESUMO
BACKGROUND: Multidrug resistance is a major problem in the treatment of breast cancer, and a number of studies have attempted to find an efficient strategy with which to overcome it. In this study, we investigate the synergistic anticancer effects of resveratrol (RSV) and doxorubicin (Dox) against human breast cancer cell lines. METHODS: The synergistic effects of RSV on chemosensitivity were examined in Dox-resistant breast cancer (MCF-7/adr) and MDA-MB-231 cells. In vivo experiments were performed using a nude mouse xenograft model to investigate the combined sensitization effect of RSV and Dox. RESULTS AND CONCLUSION: RSV markedly enhanced Dox-induced cytotoxicity in MCF-7/adr and MDA-MB-231 cells. Treatment with a combination of RSV and Dox significantly increased the cellular accumulation of Dox by down-regulating the expression levels of ATP-binding cassette (ABC) transporter genes, MDR1, and MRP1. Further in vivo experiments in the xenograft model revealed that treatment with a combination of RSV and Dox significantly inhibited tumor volume by 60%, relative to the control group. GENERAL SIGNIFICANCE: These results suggest that treatment with a combination of RSV and Dox would be a helpful strategy for increasing the efficacy of Dox by promoting an intracellular accumulation of Dox and decreasing multi-drug resistance in human breast cancer cells.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Apoptose/efeitos dos fármacos , Neoplasias da Mama/tratamento farmacológico , Resistência a Múltiplos Medicamentos/efeitos dos fármacos , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Subfamília B de Transportador de Cassetes de Ligação de ATP , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Animais , Protocolos de Quimioterapia Combinada Antineoplásica/farmacocinética , Western Blotting , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Proliferação de Células/efeitos dos fármacos , Doxorrubicina/administração & dosagem , Sinergismo Farmacológico , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Associadas à Resistência a Múltiplos Medicamentos/genética , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo Real , Resveratrol , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Estilbenos/administração & dosagem , Distribuição Tecidual , Células Tumorais CultivadasRESUMO
The uptake of nanoparticles by aquatic organisms such as fish has raised concerns about the possible adverse effects of nanoparticles (NPs). In this study, we aimed to evaluate the toxicological effects in juvenile common carp exposed to zinc oxide nanoparticles (ZnO-NPs) for 12 weeks. The carp were exposed to 0 (control), 0.1, 0.3, 0.8, and 2.4mg/L of ZnO-NPs under a flow-through exposure system. Fish were sampled at 0, 4, 8, and 12 weeks to test for zinc in the test water and blood, and biochemistry analysis; further, they were sampled at 12 weeks to observe ultrastructural changes in the liver, kidney, and gill. In the organic serum, changes in the glutamic pyruvic transaminase/alanine aminotransferase (GPT/ALT) and glutamic oxaloacetic transaminase/aspartate aminotransferase (GOT/AST) levels were significant, but changes in the lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) levels were not significantly different across all exposure periods. In the inorganic serum, the magnesium (Mg), inorganic phosphorus (IP), sodium (Na(+)), and chloride (Cl(-)) levels were significantly different in the exposure group and across exposure periods. However, calcium (Ca) and potassium (K(+)) levels were not significantly different. In the enzyme serum, the glucose (GLU) level significantly increased for the highest exposure group, but the total cholesterol (TCHO), triglyceride (Tg), and total protein (TP) levels were not significantly different during the exposure period. Ultrastructural changes in the liver induced changes in the black granules (of various sizes) in the lysosomes, indistinct nucleus membrane, and non-spherical nucleus. In the kidney, some mild changes were observed in the size and number of the lysosomes in the renal tubule. Desquamation and hypertrophy of pavement epithelial cells and vacuolation in the cytoplasm of the chloride cells were observed in the gill. Nanoparticles were also observed in the red blood cells, cytoplasm of all tissues, and glomerulus of the kidney. The observed changes in the serum and tissues may provide useful information regarding environmental conditions and risk assessments of aquatic organisms.
Assuntos
Carpas/fisiologia , Enzimas/sangue , Brânquias/efeitos dos fármacos , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Nanopartículas/toxicidade , Óxido de Zinco/toxicidade , Animais , Análise Química do Sangue , Ativação Enzimática/efeitos dos fármacos , Poluentes Químicos da Água/toxicidadeRESUMO
Ample evidence demonstrates that α-synuclein (α-syn) has a critical role in the pathogenesis of Parkinson's disease (PD) with evidence indicating that its propagation from one area of the brain to others may be the primary mechanism for disease progression. Uric acid (UA), a natural antioxidant, has been proposed as a potential disease modifying candidate in PD. In the present study, we investigated whether UA treatment modulates cell-to-cell transmission of extracellular α-syn and protects dopaminergic neurons in the α-syn-enriched model. In a cellular model, UA treatment decreased internalized cytosolic α-syn levels and neuron-to-neuron transmission of α-syn in donor-acceptor cell models by modulating dynamin-mediated and clathrin-mediated endocytosis. Moreover, UA elevation in α-syn-inoculated mice inhibited propagation of extracellular α-syn which decreased expression of phosphorylated α-syn in the dopaminergic neurons of the substantia nigra leading to their increased survival. UA treatment did not lead to change in markers related with autophagolysosomal and microglial activity under the same experimental conditions. These findings suggest UA may control the pathological conditions of PD via additive mechanisms which modulate the propagation of α-syn.
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Significant improvement in targeted therapy for colorectal cancer (CRC) has occurred over the past few decades since the approval of the EGFR inhibitor cetuximab. However, cetuximab is used only for patients possessing the wild-type oncogene KRAS, NRAS, and BRAF, and even most of these eventually acquire therapeutic resistance, via activation of parallel oncogenic pathways such as RAS-MAPK or PI3K/Akt/mTOR. The two aforementioned pathways also contribute to the development of therapeutic resistance in CRC patients, due to compensatory and feedback mechanisms. Therefore, combination drug therapies (versus monotherapy) targeting these multiple pathways may be necessary for further efficacy against CRC. In this study, we identified PIK3CA mutant (PIK3CA MT) as a determinant of resistance to SMI-4a, a highly selective PIM1 kinase inhibitor, in CRC cell lines. In CRC cell lines, SMI-4a showed its effect only in PIK3CA wild type (PIK3CA WT) cell lines, while PIK3CA MT cells did not respond to SMI-4a in cell death assays. In vivo xenograft and PDX experiments confirmed that PIK3CA MT is responsible for the resistance to SMI-4a. Inhibition of PIK3CA MT by PI3K inhibitors restored SMI-4a sensitivity in PIK3CA MT CRC cell lines. Taken together, these results demonstrate that sensitivity to SMI-4a is determined by the PIK3CA genotype and that co-targeting of PI3K and PIM1 in PIK3CA MT CRC patients could be a promising and novel therapeutic approach for refractory CRC patients.
Assuntos
Neoplasias do Colo , Fosfatidilinositol 3-Quinases , Humanos , Cetuximab/farmacologia , Cetuximab/uso terapêutico , Fosfatidilinositol 3-Quinases/genética , Neoplasias do Colo/tratamento farmacológico , Neoplasias do Colo/genética , Biomarcadores , Classe I de Fosfatidilinositol 3-Quinases/genética , Proteínas Proto-Oncogênicas c-pim-1/genéticaRESUMO
Melamine-induced renal toxicity is associated with crystal formation in the kidney following exposure to melamine and cyanuric acid. However, metabolomic profiling of intact kidney tissue after chronic intake of melamine and cyanuric acid (M + CA) mixtures has rarely been studied. The present study investigated the melamine-induced renal toxicity by determining metabolites in the kidney through [(1)H]nuclear magnetic resonance. Melamine (63 mg/kg) and cyanuric acid (6.3 mg/kg) were co-administered to rats via oral gavage for 30 days. The mixture of M + CA (63/6.3 mg/kg) induced nephrotoxicity, as determined by increased blood urea nitrogen (BUN) and creatinine levels. The kidney weights were significantly increased in the animals treated with M + CA (63/6.3 mg/kg). The histological analysis revealed epithelial degeneration and necrotic cell death in the proximal and distal tubules. Furthermore, various metabolites were altered in both renal medullar and cortical tissues. In the medullar tissues, asparagine, choline, creatinine, cysteine, ethanolamine, glucose, isoleucine, glutamine, and myo-inositol levels were elevated, but glucitol, phenylalanine, tyrosine, and sn-glycero-3-levels were reduced. In the cortex, ethanolamine, hypoxanthine, isoleucine and o-phosphoethanolamine levels were increased, whereas formate, glucose, glutathione, threonine, and myo-inositol levels were decreased, suggesting the M + CA-induced renal cell injury. These data suggest that a mixture of M + CA-induced metabolites may be useful biomarkers for the detection of kidney injury.
Assuntos
Biomarcadores/sangue , Biomarcadores/urina , Nefropatias/induzido quimicamente , Nefropatias/patologia , Metabolômica/métodos , Triazinas/toxicidade , Animais , Análise Química do Sangue , Western Blotting , Peso Corporal/efeitos dos fármacos , Imuno-Histoquímica , Rim/patologia , Espectroscopia de Ressonância Magnética , Masculino , Análise Multivariada , Tamanho do Órgão/efeitos dos fármacos , Análise de Componente Principal , Ratos , Ratos Sprague-Dawley , UrináliseRESUMO
We demonstrate significantly improved performance of a nonvolatile polymeric ferroelectric field effect transistor (FeFET) memory using nanoscopic confinement of poly(vinylidene fluoride-co-trifluoroethylene) (PVDF-TrFE) within self-assembled organosilicate (OS) lamellae. Periodic OS lamellae with 30 nm in width and 50 nm in periodicity were templated using block copolymer self-assembly. Confined crystallization of PVDF-TrFE not only significantly reduces gate leakage current but also facilitates ferroelectric polarization switching. These benefits are due to the elimination of structural defects and the development of an effective PVDF-TrFE crystal orientation through nanoconfinement. A bottom gate FeFET fabricated using a single-crystalline triisopropylsilylethynyl pentacene channel and PVDF-TrFE/OS hybrid gate insulator shows characteristic source-drain current hysteresis that is fully saturated at a programming voltage of ±8 V with an ON/OFF current ratio and a data retention time of approximately 10(2) and 2 h, respectively.
RESUMO
1,2,3-benzotriazole (BT) is used in large amounts around the world and is one of the substances derived from household chemicals that are of concern for risk when discharged to aquatic environments. Therefore, several studies have been conducted on the aquatic toxicity effects of BT, but the chronic impact assessment studies to evaluate the developmental effects on the early-life stage of fish are insufficient. In this study, the acute toxicity test and subchronic toxicity test (fish, early-life stage toxicity test, ELS test) using embryos of Japanese medaka (Oryzias latipes) were performed to evaluate the acute toxicity, developmental toxicity, growth (indicated by total length and weight at the end of the test), and histopathological effect of BT. In the short-term toxicity test on embryo and sac-fry stage, toxicity value was calculated to be 41 mg/L (NOEC). Based on this value, the exposure concentration of the ELS test was determined as 0.04, 0.4, 4 and 40 mg/L, and total exposure duration was 42 days. At the highest concentration group (40 mg/L), failure of swim bladder inflation and decrease of survival and size (total length and weight) were observed. Moreover, in the histopathological analysis, abnormal findings were detected in swim bladders from the 40 mg/L group such as inflammation and tumor changes. On the other hands, condition index (weight-length relationships, CI) was statistically significantly lower in all exposed groups compared to the control group. NOEC for the survival of BT was calculated to be 4 mg/L. LOEC for CI was 0.04 mg/L, which means BT inhibited weight gain relative to its length on larvae of medaka.
Assuntos
Oryzias , Poluentes Químicos da Água , Animais , Embrião não Mamífero , Testes de Toxicidade Aguda , Triazóis , Poluentes Químicos da Água/toxicidadeRESUMO
BACKGROUND: Mesenchymal stem cells (MSCs) may be one of candidates for disease-modifying therapy in Parkinsonian diseases. As knowledge regarding the therapeutic properties of MSCs accumulates, some obstacles still remain to be overcome, especially, successful clinical translation requires the development of culture systems that mimic the natural MSC niche, while allowing clinical-scale cell expansion without compromising quality and function of the cells. In recent years, priming approaches using bioactive peptide or complement components have been investigated to enhance the therapeutic potential of MSCs. METHODS: We investigated an innovative priming strategy by conditioning the MSCs with α-synuclein (α-syn). To induce priming, MSCs were treated with different concentrations of α-syn and various time course. We evaluated whether α-syn enhances stemness properties of MSCs and priming MSCs with α-syn would modulate autophagy-related gene expression profiles. RESULTS: Treatment of naïve MSCs with α-syn upregulated transcriptional factors responsible for regulation of stemness, which was associated with the elevated expression of genes involved in glycolysis and cell re-programming. Primed MSCs with α-syn enhanced the expression of autophagy-regulating miRNA, and exosomes derived from primed MSCs were packed with autophagy-associated miRNA. In α-syn-overexpressing neuronal cells, primed MSCs with α-syn enhanced neuronal viability relative to naïve MSCs, through the induction of autophagy and lysosome activity. Animal study using an α-syn-overexpressing mice showed that the pro-survival effect of MSCs on dopaminergic neurons was more prominent in primed MSC-treated mice compared with that in naïve MSC-treated mice. CONCLUSIONS: The present data suggest that MSC priming with α-syn exerts neuroprotective effects through augmented stemness and possibly the enhancement of autophagy-mediated α-syn modulation in Parkinsonian models.
Assuntos
Células-Tronco Mesenquimais , MicroRNAs , Fármacos Neuroprotetores , Animais , Autofagia/genética , Neurônios Dopaminérgicos/metabolismo , Células-Tronco Mesenquimais/metabolismo , Camundongos , MicroRNAs/genética , MicroRNAs/metabolismo , Fármacos Neuroprotetores/farmacologia , alfa-Sinucleína/genética , alfa-Sinucleína/metabolismo , alfa-Sinucleína/farmacologiaRESUMO
Emerging contaminants (EC) such as benzotriazole are being released into the environment in various ways, therefore it is necessary to understand how organisms are affected by EC. In this study, we exposed medaka (Oryzias latipes) and zebrafish (Danio rerio) during their embryonic period (1 day after hatching) to benzotriazole to investigate its effects on oxidative stress (ROS, GSH, GST, SOD, CAT and MDA) and changes in gene expression patterns. In both medaka and zebrafish, the influence of oxidative stress was confirmed through an increased MDA level and changes in the ROS and GSH levels. Antioxidant enzymes such as GST, CAT, and SOD were affected by benzotriazole; however, medaka and zebrafish showed different patterns in the effects by benzotriazole. Results of oxidative stress genes expression showed that medaka had either no influence or had a decrease in the gene expression profile, whereas zebrafish had a statistically significant increase in the expression of some genes. The cyp1a gene expression was increased in both species. However, vtg gene expression was increased only in zebrafish but decreased in medaka, indicating no estrogenic effects in medaka. Apoptosis genes showed changes in expression in both the species but was these changes were not dose-dependent. However, zebrafish caspase-9 gene expression was increased in all of the exposed groups, suggesting the effects on the intrinsic pathway associated with caspase-9. In conclusion, the results indicate that the toxic effects of benzotriazole differ at various levels in the two small fish medaka and zebrafish embryos.
Assuntos
Embrião não Mamífero/efeitos dos fármacos , Oryzias/embriologia , Estresse Oxidativo/efeitos dos fármacos , Triazóis/toxicidade , Peixe-Zebra/embriologia , Animais , Biomarcadores/metabolismo , Caspase 9/genética , Caspase 9/metabolismo , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Testes de Toxicidade , Vitelogeninas/genética , Vitelogeninas/metabolismo , Poluentes Químicos da Água/toxicidadeRESUMO
Background: Adult neurogenesis is the process of generating new neurons to enter neural circuits and differentiate into functional neurons. However, it is significantly reduced in Parkinson's disease (PD). Uric acid (UA), a natural antioxidant, has neuroprotective properties in patients with PD. This study aimed to investigate whether UA would enhance neurogenesis in PD. Methods: We evaluated whether elevating serum UA levels in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced parkinsonian mouse model would restore neurogenesis in the subventricular zone (SVZ). For a cellular model, we primary cultured neural precursor cells (NPCs) from post-natal day 1 rat and evaluated whether UA treatment promoted cell proliferation against 1-methyl-4-phenylpyridinium (MPP+). Results: Uric acid enhanced neurogenesis in both in vivo and in vitro parkinsonian model. UA-elevating therapy significantly increased the number of bromodeoxyuridine (BrdU)-positive cells in the SVZ of PD animals as compared to PD mice with normal UA levels. In a cellular model, UA treatment increased the expression of Ki-67. In the process of modulating neurogenesis, UA elevation up-regulated the expression of mitochondrial fusion markers. Conclusion: In MPTP-induced parkinsonian model, UA probably enhanced neurogenesis via regulating mitochondrial dynamics, promoting fusion machinery, and inhibiting fission process.
RESUMO
Ample evidence has demonstrated that α-Synuclein can propagate from one area of the brain to others via cell-to-cell transmission, which might be the underlying mechanism for pathological propagation and the disease progression of Parkinson's disease (PD). Recent reports have demonstrated cell surface receptor-mediated cell-to-cell transmission of α-synuclein. Memantine decreased the levels of internalized cytosolic α-synuclein and led to attenuation in α-synuclein-induced cell death. Specifically, memantine attenuated α-synuclein-induced expression of clathrin and EEA1, and increased expression of NR2A subunits. Moreover, memantine inhibited propagation of extracellular α-synuclein and thus, decreased the expression of the phosphorylated form of α-synuclein in dopaminergic neurons of the substantia nigra, which was accompanied by increased survival of dopaminergic neurons with functional improvement of motor deficits. The present study demonstrated that memantine modulates extracellular α-synuclein propagation by inhibiting interactions between α-synuclein and NR2A subunits, which leads to neuroprotective effects on nigral dopaminergic neurons against α-synuclein-enriched conditions. The repositioning use of memantine in α-synuclein propagation needs to be further evaluated in patients with α-synucleinopathies as an effective therapeutic approach.
Assuntos
Memantina/farmacologia , Neurônios/efeitos dos fármacos , Fármacos Neuroprotetores/farmacologia , Transtornos Parkinsonianos/patologia , alfa-Sinucleína/efeitos dos fármacos , Animais , Linhagem Celular , Humanos , Camundongos Endogâmicos C57BL , Neurônios/metabolismo , Neurônios/patologia , Transtornos Parkinsonianos/metabolismo , Receptores de N-Metil-D-Aspartato/efeitos dos fármacos , Receptores de N-Metil-D-Aspartato/metabolismo , alfa-Sinucleína/metabolismoRESUMO
A series of 3-aryl-3-azolylpropan-1-amines was prepared and screened for its capability of inhibiting monoamine reuptake. Analogs with nanomolar potency, good human in vitro microsomal stability, and low drug-drug interaction potential were described. In vivo models were used to evaluate the compound 19r for antidepressive, anxiolytic, and analgesic activity.
Assuntos
Antidepressivos/uso terapêutico , Dopamina/metabolismo , Inibidores da Captação de Neurotransmissores/uso terapêutico , Norepinefrina/metabolismo , Propilaminas/uso terapêutico , Serotonina/metabolismo , Tetrazóis/uso terapêutico , Analgésicos/química , Analgésicos/farmacocinética , Analgésicos/farmacologia , Analgésicos/uso terapêutico , Animais , Ansiolíticos/química , Ansiolíticos/farmacocinética , Ansiolíticos/farmacologia , Ansiolíticos/uso terapêutico , Antidepressivos/química , Antidepressivos/farmacocinética , Antidepressivos/farmacologia , Células CHO , Linhagem Celular , Cricetinae , Cricetulus , Humanos , Camundongos , Inibidores da Captação de Neurotransmissores/química , Inibidores da Captação de Neurotransmissores/farmacocinética , Inibidores da Captação de Neurotransmissores/farmacologia , Propilaminas/química , Propilaminas/farmacocinética , Propilaminas/farmacologia , Ratos , Tetrazóis/química , Tetrazóis/farmacocinética , Tetrazóis/farmacologiaRESUMO
Metformin has been treated for diabetes (type 2). Nowadays, this compound is frequently found in ambient water, influent/effluent of a wastewater treatment plant. To evaluate the metformin aquatic toxicity under a multi-generational exposure regimen, we exposed Oryzias latipes to metformin for two generations (133 d) and investigated its adverse effects. In the F0 generation, metformin significantly elevated gene expression for cytochrome P450 19a (CYP19a) and estrogen receptor α (ERα) in male fish; in female fish, the treatment decreased gene expression of vitellogenin (VTG2) and ERß1, suggesting endocrine disruption (one-way ANOVA, p < 0.05). Intersex occurrence of F0 female fish were found in a concentration-dependent manner, whereas no significant changes in fecundity and hatching rate were observed (p < 0.05). Metformin increased the reactive oxygen species (ROS) content, and decreased the glutathione (GSH) content in F0 male fish compared with those of the control (one-way ANOVA, p > 0.05). In F0 female fish, metformin increased catalase activity compared with that of the control (p > 0.05). The results demonstrated that metformin leads to oxidative stress and two-generation endocrine disruption in O. latipes. These results may be useful for better understanding metformin toxicity mechanism.
Assuntos
Disruptores Endócrinos/toxicidade , Hipoglicemiantes/toxicidade , Metformina/toxicidade , Oryzias/genética , Oryzias/metabolismo , Poluentes Químicos da Água/toxicidade , Animais , Catalase/metabolismo , Família 19 do Citocromo P450/genética , Transtornos do Desenvolvimento Sexual , Receptor alfa de Estrogênio/genética , Receptor beta de Estrogênio/genética , Feminino , Proteínas de Peixes/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Glutationa/metabolismo , Glutationa Transferase/metabolismo , Masculino , Estresse Oxidativo/efeitos dos fármacos , Espécies Reativas de Oxigênio/metabolismo , Vitelogeninas/genéticaRESUMO
Influenza A virus carries eight negative single-stranded RNAs and uses spliced mRNAs to increase the number of proteins produced from them. Several genome-wide screens for essential host factors for influenza A virus replication revealed a necessity for splicing and splicing-related factors, including Cdc-like kinase 1 (CLK1). This CLK family kinase plays a role in alternative splicing regulation through phosphorylation of serine-arginine rich (SR) proteins. To examine the influence that modulation of splicing regulation has on influenza infection, we analyzed the effect of CLK1 knockdown and inhibition. CLK1 knockdown in A549â¯cells reduced influenza A/WSN/33 virus replication and increased the level of splicing of segment 7, which encodes the viral M1 and M2 proteins. CLK1-/- mice infected with influenza A/England/195/2009 (H1N1pdm09) virus supported lower levels of virus replication than wild-type mice. Screening of newly developed CLK inhibitors revealed several compounds that have an effect on the level of splicing of influenza A gene segment M in different models and decrease influenza A/WSN/33 virus replication in A549â¯cells. The promising inhibitor KH-CB19, an indole-based enaminonitrile with unique binding mode for CLK1, and its even more selective analogue NIH39 showed high specificity towards CLK1 and had a similar effect on influenza mRNA splicing regulation. Taken together, our findings indicate that targeting host factors that regulate splicing of influenza mRNAs may represent a novel therapeutic approach.
Assuntos
Processamento Alternativo , Vírus da Influenza A/fisiologia , Infecções por Orthomyxoviridae/virologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Tirosina Quinases/metabolismo , RNA Mensageiro/metabolismo , Processamento Alternativo/efeitos dos fármacos , Animais , Antivirais/farmacologia , Linhagem Celular , Interações Hospedeiro-Patógeno , Humanos , Vírus da Influenza A/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Inibidores de Proteínas Quinases/farmacologia , Proteínas Serina-Treonina Quinases/antagonistas & inibidores , Proteínas Serina-Treonina Quinases/deficiência , Proteínas Serina-Treonina Quinases/genética , Proteínas Tirosina Quinases/antagonistas & inibidores , Proteínas Tirosina Quinases/deficiência , Proteínas Tirosina Quinases/genética , Fatores de Processamento de Serina-Arginina/genética , Fatores de Processamento de Serina-Arginina/metabolismo , Proteínas Virais/genética , Replicação Viral/efeitos dos fármacosRESUMO
The occurrence and distribution of hexabromocyclododecanes (HBCDs) were investigated in freshwater, sediment, and selected crucian carp (Carassius carassius) tissues (muscle, liver, egg, and blood) to evaluate the potential for HBCDs bioaccumulation. The HBCDs concentration ranged from not detected to 0.35ng/L in freshwater, and from 0.037 to 35.4ng/g-dw in sediment. The highest HBCDs concentration was detected in crucian carp liver (5.14±8.15ng/g-ww), followed by egg (3.88±10.1ng/g-ww), blood (0.61±0.63ng/mL), and muscle (0.38±0.70ng/g-ww). In all crucian carp tissues, α-HBCD was the predominant stereoisomer, and the fraction of α-HBCD as a proportion of the total HBCDs in liver tissue (96%) was higher than that in egg tissue (79%). There was a positive correlation between the HBCDs concentration in crucian carp muscle and body size (p<0.01, Spearman). The biota-sediment accumulation factor (BSAF) (0.14) and bioconcentration factor (BCF) (137,000L/kg) values were estimated in crucian carp muscle using field-based data.
Assuntos
Carpas/metabolismo , Monitoramento Ambiental/métodos , Hidrocarbonetos Bromados/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Água Doce/químicaRESUMO
Zinc oxide nanoparticles (ZnO NPs) and titanium dioxide nanoparticles (TiO2 NPs) are well known as photoreactive nanoparticles (NPs). Various phototoxicities of ZnO NPs and TiO2 NPs were reported on several organisms. It was still necessary to evaluate the toxicity of photoreactive ZnO NPs and TiO2 NPs due to species-specific effects under various irradiation conditions. We compared the acute toxicity of Moina macrocopa under visible, ultraviolet (UV) A, and B irradiations, according to the Organization for Economic Cooperation and Development guidelines for the testing of chemicals (Test No. 202). The sensitivity of ZnO NPs for M. macrocopa was UVB>UVA>visible light irradiation. There were no significant lethal and immobile effects of TiO2 NPs on juveniles under all irradiations and in the tested concentrations of TiO2 NPs. Photoreactive NPs have a potential and accelerated toxicity on organisms in the ambient environments.
RESUMO
To elucidate the multi-generational estrogenic potential of Perfluoroalkyl acids (PFAAs) mixture, vitellogenin (VTG) expression, growth indices, histological alteration, fecundity, hatching rate, larval survival rate, and sex ratio of Japanese medaka (Oryzias latipes) were investigated by exposing the fish to a mixture of perfluorooctane sulfonate (PFOS), perfluroroctanoic acid (PFOA), perfluorobutane sulfonate (PFBS), and perfluorononanoic acid (PFNA) for three generations (238 days). Mixture composition is in the ratio of 1:1:1:1. In addition, whole body burden for each PFAA was analyzed. According to the results, concentrated levels of the PFAAs in both F1 and F2 generation O. latipes were ordered PFOS > PFNA > PFOA > PFBS at both low concentration (0.5 µg/L) and high concentration (5 µg/L), whereas a significant difference in whole body burden based on sex or generation was not detected. Significant induction of VTG expression in F2 and the decline of the gonad somatic index (GSI) in F1 were observed following PFAAs mixture exposure (p < 0.05, one-way ANOVA). Furthermore, suppression level of reproduction rate relative to the control increased as generation was transferred to the next in response to PFAAs mixture or 17 ß-estradiol exposure, with the inhibition of hatchability observed in the F1 generation. The PFAA high concentration caused significant alteration of F1 generation sex ratio, suggesting the adverse effect of PFAA in population level (Chi-square test, P > 0.05). Overall, this study demonstrated that PFAA mixture could have the potential of multi-generational endocrine disruptors in O. latipes.