RESUMEN

Castor (Ricinus communis) is a relevant industrial oilseed feedstock for many industrial applications, being globally mainly cultivated by smallholder farmers in semiarid areas, where abiotic stresses predominate. Therefore, susceptible to generating reactive oxygen species (ROS) and subsequent oxidative stress, compromising cell metabolism upon seed imbibition and germination, seedling and crop establishment, and yield. The present study evaluated the consequences of water restriction by Polyethylene glycol (PEG) and Sodium chloride (NaCl) on cell cycle and metabolism reactivation on germinability, seedling growth, and vigor parameters in 2 commercial castor genotypes (Nordestina and Paraguaçu). PEG water restriction inhibited germination completely at -0.23 MPa or higher, presumably due to reduced oxygen availability. The restrictive effects of NaCl saline stress on germination were observed only from -0.46 MPa onwards, affecting dry mass accumulation and the production of normal seedlings. In general, superoxide dismutase (SOD) activity increased in NaCl -0.23 MPa, whereas its modulation during the onset of imbibition (24h) seemed to depend on its initial levels in dry seeds in a genotype-specific manner, therefore, resulting in the higher stress tolerance of Nordestina compared to Paraguaçu. Overall, results show that Castor germination and seedling development are more sensitive to the restrictive effects of PEG than NaCl at similar osmotic potentials, contributing to a better understanding of the responses to water restriction stresses by different Castor genotypes. Ultimately, SOD may constitute a potential marker for characterizing castor genotypes in stressful situations during germination, early seedling, and crop establishment, and a target for breeding for Castor-improved stress tolerance.

Asunto(s)

Ricinus communis , Plantones , Plantones/metabolismo , Cloruro de Sodio/farmacología , Cloruro de Sodio/metabolismo , Ricinus communis/genética , Polietilenglicoles/farmacología , Polietilenglicoles/metabolismo , Germinación , Ciclo Celular , Semillas/metabolismo , Agua/metabolismo , Superóxido Dismutasa/metabolismo

RESUMEN

PURPOSE: Liver damage caused by drugs and other chemicals accounts for about 5% of all cases. Methotrexate (MTX), a folic acid analogue, is a first-line synthetic antimetabolite agent routinely used in the treatment of rheumatoid arthritis and other autoimmune and chronic inflammatory diseases. Polyethylene glycol (PEG) has antioxidant activity. In this study, we evaluated biochemically and histopathologically the antifibrotic effect of PEG 3350 administered intraperitoneally to prevent methotrexate-induced liver damage in rats. METHODS: A total of 30 male rats including 10 rats was given no drugs (normal group), and 20 rats received single-dose 20 mg/kg MTXfor induced liver injury in this study. MTX was given to 20 rats, which were divided in two groups. Group 1 rats was given PEG30 mg/kg/day (Merck) intraperitoneally, and Group 2 rats % 0.9 NaCl saline 1 mL/kg/day intraperitoneally daily for two weeks. RESULTS: Transforming growth factor beta (TGF-ß), plasma malondialdehyde (MDA), liver MDA, serum tumour necrosis factor alpha (TNF-α), alanine aminotransferase and plasma pentraxin-3 levels and, according to tissue histopathology, hepatocyte necrosis, fibrosis and cellular infiltration were significantly better in MTX+PEG group than in MTX+saline group. CONCLUSIONS: PEG 3350 is a hope for toxic hepatitis due to other causes, since liver damage occurs through oxidative stress and cell damage, similar to all toxic drugs.

Asunto(s)

Hepatopatías , Metotrexato , Animales , Masculino , Malondialdehído/metabolismo , Metotrexato/efectos adversos , Estrés Oxidativo , Polietilenglicoles/metabolismo , Polietilenglicoles/farmacología , Ratas , Ratas Wistar

RESUMEN

Cytochrome c (Cyt-c), a small mitochondrial electron transport heme protein, has been employed in bioelectrochemical and therapeutic applications. However, its potential as both a biosensor and anticancer drug is significantly impaired due to poor long-term and thermal stability. To overcome these drawbacks, we developed a site-specific PEGylation protocol for Cyt-c. The PEG derivative used was a 5 kDa mPEG-NHS, and a site-directed PEGylation at the lysine amino-acids was performed. The effects of the pH of the reaction media, molar ratio (Cyt-c:mPEG-NHS) and reaction time were evaluated. The best conditions were defined as pH 7, 1:25 Cyt-c:mPEG-NHS and 15 min reaction time, resulting in PEGylation yield of 45% for Cyt-c-PEG-4 and 34% for Cyt-c-PEG-8 (PEGylated cytochrome c with 4 and 8 PEG molecules, respectively). Circular dichroism spectra demonstrated that PEGylation did not cause significant changes to the secondary and tertiary structures of the Cyt-c. The long-term stability of native and PEGylated Cyt-c forms was also investigated in terms of peroxidative activity. The results demonstrated that both Cyt-c-PEG-4 and Cyt-c-PEG-8 were more stable, presenting higher half-life than unPEGylated protein. In particular, Cyt-c-PEG-8 presented great potential for biomedical applications, since it retained 30-40% more residual activity than Cyt-c over 60-days of storage, at both studied temperatures of 4 °C and 25 °C.

Asunto(s)

Citocromos c , Lisina , Dicroismo Circular , Citocromos c/química , Lisina/química , Polietilenglicoles/química , Polietilenglicoles/metabolismo , Polietilenglicoles/farmacología , Proteínas

RESUMEN

PURPOSE: The aim of this work was to formulate and characterize surfactant-free glibenclamide nanoparticles using Eudragit RLPO and polyethylene glycol as sole stabilizer. METHODS: Glibenclamide nanoparticles were obtained by nanoprecipitation and evaluated in terms of drug content, encapsulation efficiency, apparent saturation solubility, drug release profile, solid state and storage stability. The influence of different stirring speed on the particle size, size distribution and zeta potential of the nanoparticles was investigated. The nanoparticle biocompatibility and permeability were analyzed in vitro on Caco-2 cell line (clone HTB-37) and its interaction with mucin was also investigated. RESULTS: It was found that increasing the molecular weight of polyethylene glycol from 400 to 6000 decreased drug encapsulation, whereas the aqueous solubility and dissolution rate of the drug increased. Particle size of the nanoformulations, with and without polyethylene glycol, were between 140 and 460 nm. Stability studies confirmed that glibenclamide nanoparticles were stable, in terms of particle size, after 120 days at 4°C. In vitro studies indicated minimal interactions of glibenclamide nanoparticles and mucin glycoproteins suggesting favorable properties to address the intestinal mucus barrier. Cell viability studies confirmed the safety profile of these nanoparticles and showed an increased permeation through epithelial cells. CONCLUSION: Taking into consideration these findings, polyethylene glycol is a useful polymer for stabilizing these surfactant-free glibenclamide nanoparticles and represent a promising alternative to improve the treatment of non-insulin dependent diabetes.

Asunto(s)

Composición de Medicamentos/métodos , Gliburida/metabolismo , Hipoglucemiantes/metabolismo , Mucosa Intestinal/metabolismo , Nanopartículas/metabolismo , Tensoactivos , Células CACO-2 , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/fisiología , Evaluación Preclínica de Medicamentos/métodos , Gliburida/administración & dosificación , Gliburida/química , Humanos , Hipoglucemiantes/administración & dosificación , Hipoglucemiantes/química , Mucosa Intestinal/efectos de los fármacos , Nanopartículas/administración & dosificación , Nanopartículas/química , Tamaño de la Partícula , Polietilenglicoles/administración & dosificación , Polietilenglicoles/química , Polietilenglicoles/metabolismo , Polímeros/administración & dosificación , Polímeros/química , Polímeros/metabolismo

RESUMEN

Simple size observations of 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethyleneglycol)-2000] (DSPE-mPEG2000) polymeric micelles (PM) with different compositions including or not paclitaxel (PTX) are unable to evidence changes on the nanocarrier structure. In such system a detailed characterization using highly sensitive techniques such as X-ray scattering and asymmetric flow field flow fractionation coupled to multi-angle laser light scattering and dynamic light scattering (AF4-MALS-DLS) is mandatory to observe effects that take place by the addition of PTX and/or more lipid-polymer at PM, leading to complex changes on the structure of micelles, as well as in their supramolecular organization. SAXS and AF4-MALS-DLS suggested that PM can be found in the medium separately and highly organized, forming clusters of PM in the latter case. SAXS fitted parameters showed that adding the drug does not change the average PM size since the increase in core radius is compensated by the decrease in shell radius. SAXS observations indicate that PEG conformation takes place, changing from brush to mushroom depending on the PM composition. These findings directly reflect in in vivo studies of blood clearance that showed a longer circulation time of blank PM when compared to PM containing PTX.

Asunto(s)

Paclitaxel/sangre , Fosfatidiletanolaminas/sangre , Polietilenglicoles/metabolismo , Animales , Cápsulas/química , Cápsulas/metabolismo , Ratones , Micelas , Estructura Molecular , Paclitaxel/química , Fosfatidiletanolaminas/química , Polietilenglicoles/química , Dispersión del Ángulo Pequeño , Difracción de Rayos X

RESUMEN

Chitosan has received a lot of attention as a carrier for small interfering RNA (siRNA), due to its capacity for complexation and intracellular release of these molecules. However, one of its limitations is its insolubility at neutral pH and the tendency towards aggregation of its nanoparticles in isotonic ionic strength. In this study, a series of amphipathic chitosans were synthesized by varying the degree of acetylation (DA) from ˜2 to ˜30 mol% and the degree of substitution (DS) from 5 to 25%. by tertiary amino groups (DEAE) The results showed that the adjustment of these parameters decreases the interparticle interactions mediated by hydrogen bonding to obtain nanoparticles with improved colloidal stability. siRNA-containing nanoparticles of 100 to 150 nm with low polydispersities (0.15-0.2) and slightly positive zeta potentials (˜+ 5 mV) were resistant to aggregation at pH 7.4 and ionic strength of 150 mM. This resistance to aggregation is provided by changes on the nanoparticle surface and highlights the importance of more organized self-assembly in providing colloidal stability at physiological conditions. Additionally, the PEGylation of the most promising vectors conferred favorable physicochemical properties to nanoparticles. The chitosans and their nanoparticles exhibited low toxicity and an efficient cell uptake, as probed by confocal microscopy of rhodamine labeled vectors. The results provide a new approach to overcome the limited stability of chitosan nanoparticles at physiological conditions and show the potential of these amphipathic chitosans as siRNA carriers.

Asunto(s)

Quitosano/análogos & derivados , Portadores de Fármacos/química , Nanopartículas/química , ARN Interferente Pequeño/administración & dosificación , Tensoactivos/química , Anhídridos Acéticos/química , Acetilación , Animales , Quitosano/síntesis química , Quitosano/metabolismo , Quitosano/toxicidad , Dietilaminas/química , Portadores de Fármacos/síntesis química , Portadores de Fármacos/metabolismo , Portadores de Fármacos/toxicidad , Fluorescencia , Colorantes Fluorescentes/química , Concentración de Iones de Hidrógeno , Ratones , Nanopartículas/metabolismo , Nanopartículas/toxicidad , Tamaño de la Partícula , Polietilenglicoles/síntesis química , Polietilenglicoles/química , Polietilenglicoles/metabolismo , Polietilenglicoles/toxicidad , Células RAW 264.7 , ARN Interferente Pequeño/química , Rodaminas/química , Tensoactivos/síntesis química , Tensoactivos/metabolismo , Tensoactivos/toxicidad

RESUMEN

L-asparaginase (ASNase) from Escherichia coli is currently used in some countries in its PEGylated form (ONCASPAR, pegaspargase) to treat acute lymphoblastic leukemia (ALL). PEGylation refers to the covalent attachment of poly(ethylene) glycol to the protein drug and it not only reduces the immune system activation but also decreases degradation by plasmatic proteases. However, pegaspargase is randomly PEGylated and, consequently, with a high degree of polydispersity in its final formulation. In this work we developed a site-specific N-terminus PEGylation protocol for ASNase. The monoPEG-ASNase was purified by anionic followed by size exclusion chromatography to a final purity of 99%. The highest yield of monoPEG-ASNase of 42% was obtained by the protein reaction with methoxy polyethylene glycol-carboxymethyl N-hydroxysuccinimidyl ester (10kDa) in 100 mM PBS at pH 7.5 and PEG:ASNase ratio of 25:1. The monoPEG-ASNase was found to maintain enzymatic stability for more days than ASNase, also was resistant to the plasma proteases like asparaginyl endopeptidase and cathepsin B. Additionally, monoPEG-ASNase was found to be potent against leukemic cell lines (MOLT-4 and REH) in vitro like polyPEG-ASNase. monoPEG-ASNase demonstrates its potential as a novel option for ALL treatment, being an inventive novelty that maintains the benefits of the current enzyme and solves challenges.

Asunto(s)

Asparaginasa/química , Asparaginasa/metabolismo , Polietilenglicoles/metabolismo , Asparaginasa/aislamiento & purificación , Asparaginasa/farmacología , Sitios de Unión , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Cromatografía en Gel , Estabilidad de Enzimas , Humanos , Leucemia-Linfoma Linfoblástico de Células Precursoras/tratamiento farmacológico

RESUMEN

The effects on the vasculature produced by ethanol withdrawal include both vasodilatation and hypocontractility, although a detailed biochemical understanding of these processes is yet to be accomplished. Here, we sought to investigate some of the mechanisms underlying vascular hypocontractility induced by ethanol withdrawal. Male Wistar rats were treated with increasing doses of 3-9% ethanol (v/v) for 21 days and the impact of ethanol withdrawal on the vascular function was assessed 48 h after immediate ethanol suspension. Endothelium-denuded rat aortic rings showed a reduced contractile response to phenylephrine, angiotensin II, serotonin and KCl after ethanol withdrawal, but the same phenomenon was not observed in endothelium-intact rings. Indomethacin, but not L-NAME, tiron, PEG-catalase and SC560, restored the contractile response to phenylephrine of endothelium-denuded aortas from abstinent rats. Hyporeactivity to phenylephrine induced by ethanol withdrawal was reversed by SC236, a selective cyclooxygenase (COX)-2 inhibitor. Similarly, Ro1138452, a selective prostacyclin IP receptor antagonist, reversed vascular hypocontractility induced by ethanol withdrawal. Increased concentrations of 6-keto-prostaglandin (PG)F1α, a stable product of PGI2, was detected in endothelium-denuded aortas from abstinent rats, and this response was prevented by indomethacin. However, no changes in aortic PGE2 levels were detected after ethanol withdrawal. In situ quantification of hydrogen peroxide (H2O2) and nitric oxide (NO) using fluorescent dyes revealed that ethanol withdrawal decreased the levels of these two compounds in the tunica media. Our studies show that the vascular hypocontractility induced by ethanol withdrawal is independent of the endothelium and it is mediated by PGI2 derived from COX-2.

Asunto(s)

Ciclooxigenasa 2/metabolismo , Epoprostenol/metabolismo , Etanol/efectos adversos , Síndrome de Abstinencia a Sustancias/metabolismo , Animales , Aorta/efectos de los fármacos , Aorta/metabolismo , Compuestos de Bencilo/farmacología , Catalasa/metabolismo , Inhibidores de la Ciclooxigenasa 2/farmacología , Endotelio Vascular/metabolismo , Peróxido de Hidrógeno/farmacología , Imidazoles/farmacología , Indometacina/farmacología , Masculino , Fenilefrina/farmacología , Polietilenglicoles/metabolismo , Pirazoles/farmacología , Ratas , Ratas Wistar , Sulfonamidas/farmacología , Vasoconstricción/efectos de los fármacos , Vasodilatación/efectos de los fármacos

RESUMEN

Crisantaspase is an asparaginase enzyme produced by Erwinia chrysanthemi and used to treat acute lymphoblastic leukemia (ALL) in case of hypersensitivity to Escherichia coli l-asparaginase (ASNase). The main disadvantages of crisantaspase are the short half-life (10 H) and immunogenicity. In this sense, its PEGylated form (PEG-crisantaspase) could not only reduce immunogenicity but also improve plasma half-life. In this work, we developed a process to obtain a site-specific N-terminal PEGylated crisantaspase (PEG-crisantaspase). Crisantaspase was recombinantly expressed in E. coli BL21(DE3) strain cultivated in a shaker and in a 2-L bioreactor. Volumetric productivity in bioreactor increased 37% compared to shaker conditions (460 and 335 U L-1  H-1 , respectively). Crisantaspase was extracted by osmotic shock and purified by cation exchange chromatography, presenting specific activity of 694 U mg-1 , 21.7 purification fold, and yield of 69%. Purified crisantaspase was PEGylated with 10 kDa methoxy polyethylene glycol-N-hydroxysuccinimidyl (mPEG-NHS) at different pH values (6.5-9.0). The highest N-terminal pegylation yield (50%) was at pH 7.5 with the lowest poly-PEGylation ratio (7%). PEG-crisantaspase was purified by size exclusion chromatography and presented a KM value three times higher than crisantaspase (150 and 48.5 µM, respectively). Nonetheless, PEG-crisantaspase was found to be more stable at high temperatures and over longer periods of time. In 2 weeks, crisantaspase lost 93% of its specific activity, whereas PEG-crisantaspase was stable for 20 days. Therefore, the novel PEG-crisantaspase enzyme represents a promising biobetter alternative for the treatment of ALL.

Asunto(s)

Asparaginasa/biosíntesis , Asparaginasa/química , Polietilenglicoles/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Asparaginasa/metabolismo , Humanos , Cinética , Polietilenglicoles/química , Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo

RESUMEN

Plants in arid zones are constantly exposed to drought stress. The ASR protein family (Abscisic, Stress, Ripening) -a subgroup of the late embryogenesis abundant superfamily- is involved in the water stress response and adaptation to dry environments. Tomato ASR1, as well as other members of this family, is an intrinsically disordered protein (IDP) that functions as a transcription factor and a chaperone. Here we employed different biophysical techniques to perform a deep in vitro characterization of ASR1 as an IDP and showed how both environmental factors and in vivo targets modulate its folding. We report that ASR1 adopts different conformations such as α-helix or polyproline type II in response to environmental changes. Low temperatures and low pH promote the polyproline type II conformation (PII). While NaCl increases PII content and slightly destabilizes α-helix conformation, PEG and glycerol have an important stabilizing effect of α-helix conformation. The binding of Zn2+in the low micromolar range promotes α-helix folding, while extra Zn2+ results in homo-dimerization. The ASR1-DNA binding is sequence specific and dependent on Zn2+. ASR1 chaperone activity does not change upon the structure induction triggered by the addition of Zn2+. Furthermore, trehalose, which has no effect on the ASR1 structure by itself, showed a synergistic effect on the ASR1-driven heat shock protection towards the reporter enzyme citrate synthase (CS). These observations prompted the development of a FRET reporter to sense ASR1 folding in vivo. Its performance was confirmed in Escherichia coli under saline and osmotic stress conditions, representing a promising probe to be used in plant cells. Overall, this work supports the notion that ASR1 plasticity is a key feature that facilitates its response to drought stress and its interaction with specific targets.

Asunto(s)

Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Solanum lycopersicum/crecimiento & desarrollo , Estrés Fisiológico , Frío , Sequías , Glicerol/metabolismo , Concentración de Iones de Hidrógeno , Solanum lycopersicum/metabolismo , Polietilenglicoles/metabolismo , Unión Proteica , Multimerización de Proteína , Estructura Secundaria de Proteína , Desplegamiento Proteico , Trehalosa/metabolismo , Zinc/metabolismo

RESUMEN

OBJECTIVE: To determine whether trace amounts of ethylene glycol (EG), diethylene glycol (DEG), or triethylene glycol (TEG) in PEG 3350 are associated with increased blood levels of EG, DEG, or TEG in children receiving daily PEG 3350 therapy. STUDY DESIGN: Blood samples were drawn from 9 children who were being treated for constipation with PEG 3350 (6-12 years old) before and every 30 minutes for 3 hours after receiving 17 g of PEG 3350. PEG 3350, tap water, and blood samples from 18 age- and sex-matched controls also were analyzed. RESULTS: Baseline blood levels of EG and TEG did not differ between control and treated groups. DEG levels (median [IQR]) were lower in the PEG 3350 group (40.13 ng/mL [36.69, 63.94] vs 92.83 ng/mL [51.06, 128.93], P = .008). After PEG 3350 dose, levels of EG (390.51 ng/mL [326.06, 624.55]) and TEG (2.21 ng/mL [0, 4.5]) peaked at 90 minutes at 1032.81 ng/mL (826.84, 1486.13) (P = .009) and 35.17 ng/mL (15.81, 45.13) (P = .0005), respectively. DEG levels did not significantly change. Standard 17-g doses of PEG 3350 in 8 oz (237 mL) of water resulted in concentrations (mean ± SD) of EG, DEG, and TEG of 1.32 ± 0.23 µg/mL, 0.18 ± 0.03 µg/mL, and 0.12 ± 0.01 µg/mL, respectively. EG, DEG, and TEG levels in public water supply were 0.07 µg/mL, 0.21 µg/mL, and 0.02 µg/mL, respectively. CONCLUSIONS: Daily PEG 3350 therapy in children was not associated with sustained elevation of EG, DEG, or TEG blood levels over levels in matched controls. Although EG and TEG levels increased after a standard dose of PEG 3350, their peak values remained well below toxic levels.

Asunto(s)

Glicol de Etileno/sangre , Glicoles de Etileno/sangre , Laxativos/química , Polietilenglicoles/química , Polietilenglicoles/metabolismo , Biomarcadores/sangre , Estudios de Casos y Controles , Niño , Estreñimiento/sangre , Estreñimiento/tratamiento farmacológico , Femenino , Humanos , Laxativos/uso terapéutico , Masculino , Polietilenglicoles/uso terapéutico

RESUMEN

BACKGROUND AND OBJECTIVE: In this mini-review, we have compiled the most recent and comparable information to shed light on the action of PEGylation in the biodistribution of carbon nanotubes (CNT) in the central nervous system (CNS). It is well known that due to the complexity of the CNS and the severity of the outcome following changes in this system, this is one of the areas where there are more investments in research to develop new technologies and approaches for more effective and less invasive treatments. The CNS is highly protected against toxic and invasive microorganisms thanks to the blood brain barrier (BBB), but this protection also prevents the passage of potentially beneficial molecules for the treatment of neurological disorders. Nanotechnology attempts to develop nanocompounds that are biocompatible and non-immunogenic, and that are able to cross the BBB in therapeutic amounts without causing damage and to diffuse through nerve tissue. These compounds should also be cleared and biodistributed properly, being capable of performing drug delivery exclusively for CNS pathologies, such as neurodegenerative diseases (Parkinson's and Alzheimer's) and brain tumors. CONCLUSION: In this way, this review focuses on CNT PEGylation, aiming to help in the development of viable and effective nanomedicines for neuroscience applications.

Asunto(s)

Enfermedades del Sistema Nervioso Central/metabolismo , Nanotubos de Carbono , Polietilenglicoles/metabolismo , Distribución Tisular/fisiología , Animales , Barrera Hematoencefálica/fisiología , Humanos , Nanotecnología

RESUMEN

Nowadays cancer is one of the most common causes of deaths worldwide. Conventional antitumor agents still present various problems related to specificity for tumor cells often leading to therapeutic failure. Nanoscale particles are considered potential alternative to direct access of drugs into tumor cells, therefore increasing the drug accumulation and performance. The aim of this study was to evaluate the antitumor activity of doxorubicin (DOX)-loaded nanostructured lipid carriers (NLC) versus liposomes against a breast cancer animal experimental model. NLC-DOX and liposomes-DOX were successfully prepared and characterized. Tumor-bearing mice were divided into five groups (blank-NLC, blank-liposome, DOX, NLC-DOX, liposome-DOX). Each animal received by the tail vein four doses of antitumoral drugs (total dose, 16mg/kg), every 3 days. Antitumor efficacy was assessed by measuring 1) tumor volume, calculating the inhibitory ratio (TV-IR, see after) and 2) acquiring scintigraphic images of the tumor using doxorubicin radiolabeled with technetium-99m as an imaging tumor probe. Liposome-DOX and free DOX did not showed differences in the tumor mean volume, whereas NLC-DOX proved to be the best treatments in controlling the tumor growth. NLC-DOX showed an inhibition ration (TV-IR) of 73.5% while free DOX and liposome-DOX decreased TV-RI of 48.8% and 68.0%, respectively. Tumor was clearly visualized in controls, DOX, and liposome-DOX groups. Yet, regarding the NLC-DOX group, tumor was barely identified by the image, indicating antitumor efficacy. Moreover, both NLC and liposomes proved to be able to delay the occurrence of lung metastasis. In conclusion, results of this study indicated that NLC-DOX might be an alternative strategy to achieve an efficient antitumor activity.

Asunto(s)

Antibióticos Antineoplásicos/administración & dosificación , Neoplasias de la Mama/tratamiento farmacológico , Doxorrubicina/análogos & derivados , Lípidos/química , Nanopartículas , Animales , Antibióticos Antineoplásicos/química , Antibióticos Antineoplásicos/metabolismo , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Línea Celular Tumoral , Doxorrubicina/administración & dosificación , Doxorrubicina/química , Doxorrubicina/metabolismo , Composición de Medicamentos , Femenino , Inyecciones Intravenosas , Liposomas , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/prevención & control , Neoplasias Pulmonares/secundario , Ratones Endogámicos BALB C , Polietilenglicoles/administración & dosificación , Polietilenglicoles/química , Polietilenglicoles/metabolismo , Factores de Tiempo , Carga Tumoral

RESUMEN

OBJECTIVES: Our main objective was to investigate the mechanisms underlying the effects of hyperhomocysteinaemia (HHcy) on contractile response mediated by α1-adrenoceptors in the rat corpus cavernosum. METHODS: Concentration-response curves for phenylephrine (PE) were obtained in strips of corpus cavernosum, in absence or after incubation with tiron, tempol or polyethylene glycol (PEG)-catalase combined or not with tempol. We also measured the superoxide anion (O2(-)) and hydrogen peroxide (H2O2) generation, superoxide dismutase (SOD) and catalase activity and α-actin expression in rat corpus cavernosum from both groups. KEY FINDINGS: HHcy increased PE-induced contraction in cavernosal strips. Tiron, PEG-catalase or tempol increased PE-induced contraction in strips from control rats, but it was not altered by tiron or PEG-catalase in HHcy rats, whereas tempol reduced this response. The combination of PEG-catalase and tempol did not alter the contractile response to PE in both groups. HHcy increased O2(-) generation and SOD activity, whereas H2O2 concentration was reduced. Finally, HHcy did not alter catalase activity or expression of α-actin. CONCLUSIONS: The major new finding from this study is that HHcy induced a marked increase in PE-induced contraction in rat corpus cavernosum by a mechanism that involves increased O2(-) generation and it could play a role in the pathogenesis of erectile dysfunction associated with HHcy.

Asunto(s)

Hiperhomocisteinemia/metabolismo , Pene/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Receptores Adrenérgicos alfa 1/metabolismo , Actinas/metabolismo , Animales , Catalasa/metabolismo , Disfunción Eréctil/metabolismo , Peróxido de Hidrógeno/metabolismo , Masculino , Contracción Muscular/efectos de los fármacos , Contracción Muscular/fisiología , Músculo Liso/efectos de los fármacos , Músculo Liso/metabolismo , Pene/efectos de los fármacos , Fenilefrina/farmacología , Polietilenglicoles/metabolismo , Ratas , Ratas Wistar , Superóxidos/metabolismo

RESUMEN

The use of oxidoredutive enzymes in removing organic pollutants has been the subject of much research. The oxidation of phenolic compounds in the presence of chemical additives has been the focus of this study. In this investigation, the influence of the additives polyethylene glycol and Triton X-100 was evaluated in the phenol oxidation, caffeic acid, chlorogenic acid and total phenolic compounds present in coffee processing wastewater (CPW) at different pH values, performed by turnip peroxidase and peroxidase extracted from soybean seed hulls. The influence of these additives was observed only in the oxidation of phenol and caffeic acid. In the oxidation of other studied phenolic compounds, the percentage of oxidation remained unchanged in the presence of these chemical additives. In the oxidation of CPW in the presence of additives, no change in the oxidation of phenolic compounds was observed. Although several studies show the importance of evaluating the influence of additives on the behaviour of enzymes, this study found a positive response from the economic point of view for the treatment of real wastewater, since the addition of these substances showed no influence on the oxidation of phenolic compounds, which makes the process less costly.

Asunto(s)

Brassica napus/enzimología , Glycine max/enzimología , Peroxidasas/metabolismo , Fenoles/aislamiento & purificación , Eliminación de Residuos Líquidos/métodos , Aguas Residuales/análisis , Ácidos Cafeicos/aislamiento & purificación , Ácidos Cafeicos/metabolismo , Ácido Clorogénico/aislamiento & purificación , Ácido Clorogénico/metabolismo , Estabilidad de Enzimas/efectos de los fármacos , Residuos Industriales/análisis , Octoxinol/metabolismo , Oxidación-Reducción , Peroxidasas/aislamiento & purificación , Fenoles/metabolismo , Polietilenglicoles/metabolismo , Sustancias Protectoras/metabolismo , Purificación del Agua/métodos

RESUMEN

Nitric oxide (NO) releasing biomaterials represent a potential strategy for use as active wound dressings capable of accelerating wound healing. Topical NO-releasing poly(vinyl alcohol) (PVA) films and Pluronic F127 hydrogels (F127) have already exhibited effective skin vasodilation and wound healing actions. In this study, we functionalized PVA films with SNO groups via esterification with a mixture of mercaptosucinic acid (MSA) and thiolactic acid (TLA) followed by S-nitrosation of the SH moieties. These films were combined with an underlying layer of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide), i.e., PEO-PPO-PEO (Pluronic F127) hydrogel and used for the topical treatment of skin lesions in an animal model. The mixed esterification of PVA with MSA and TLA led to chemically crosslinked PVA-SNO films with a high swelling capacity capable of spontaneously releasing NO. Real time NO-release measurements revealed that the hydrogel layer reduces the initial NO burst from the PVA-SNO films. We demonstrate that the combination of PVA-SNO films with F127 hydrogel accelerates wound contraction, decreases wound gap and cellular density and accelerates the inflammatory phase of the lesion. These results were reflected in an increase in myofibroblastic differentiation and collagen type III expression in the cicatricial tissue. Therefore, PVA-SNO films combined with F127 hydrogel may represent a new approach for active wound dressings capable of accelerating wound healing.

Asunto(s)

Hidrogeles/química , Óxido Nítrico/química , Poloxámero/química , Alcohol Polivinílico/química , Actinas/metabolismo , Animales , Antígenos de Diferenciación/metabolismo , Western Blotting , Hidrogeles/metabolismo , Hidrogeles/farmacología , Inmunohistoquímica , Masculino , Ratones , Óxido Nítrico/metabolismo , Poloxámero/metabolismo , Polietilenglicoles/química , Polietilenglicoles/metabolismo , Alcohol Polivinílico/metabolismo , Alcohol Polivinílico/farmacología , Glicoles de Propileno/química , Glicoles de Propileno/metabolismo , S-Nitrosoglutatión/química , S-Nitrosoglutatión/metabolismo , Piel/metabolismo , Piel/patología , Piel/fisiopatología , Compuestos de Sulfhidrilo/química , Tiomalatos/química , Factores de Tiempo , Cicatrización de Heridas/efectos de los fármacos

RESUMEN

Studies were conducted to determine the effect of osmotic and matric stress on germination and growth of two Fusarium solani strains, the etiological agent responsible of peanut brown root rot. Both strains had similar osmotic and matric potential ranges that allowed growth, being the latter one narrower. F. solani showed the ability to grow down to -14 MPa at 25 °C in non-ionic modified osmotic medium, while under matric stress this was limited to -8.4 MPa at 25 °C. However, both strains were seen to respond differently to decreasing osmotic and matric potentials, during early stages of germination. One strain (RC 338) showed to be more sensitive to matric than osmotic (non ionic) and the other one (RC 386) showed to be more sensitive to osmotic than matric imposed water stress. After 24 h of incubation, both isolates behaved similarly. The minimum water potential for germination was -8.4 MPa on glycerol amended media and -5.6 MPa for NaCl and PEG amended media, respectively. The knowledge of the water potential range which allow mycelia growth and spore germination of F. solani provides an inside to the likely behaviour of this devastating soilborne plant pathogen in nature and has important practical implications.

Asunto(s)

Fusarium/crecimiento & desarrollo , Presión Osmótica , Agua/metabolismo , Arachis/microbiología , Fusarium/efectos de los fármacos , Fusarium/efectos de la radiación , Glicerol/metabolismo , Enfermedades de las Plantas/microbiología , Polietilenglicoles/metabolismo , Cloruro de Sodio/metabolismo , Microbiología del Suelo , Temperatura

RESUMEN

Studies were conducted to determine the effect of osmotic and matric stress on germination and growth of two Fusarium solani strains, the etiological agent responsible of peanut brown root rot. Both strains had similar osmotic and matric potential ranges that allowed growth, being the latter one narrower. F. solani showed the ability to grow down to -14 MPa at 25 °C in non-ionic modified osmotic medium, while under matric stress this was limited to -8.4 MPa at 25 °C. However, both strains were seen to respond differently to decreasing osmotic and matric potentials, during early stages of germination. One strain (RC 338) showed to be more sensitive to matric than osmotic (non ionic) and the other one (RC 386) showed to be more sensitive to osmotic than matric imposed water stress. After 24 h of incubation, both isolates behaved similarly. The minimum water potential for germination was -8.4 MPa on glycerol amended media and -5.6 MPa for NaCl and PEG amended media, respectively. The knowledge of the water potential range which allow mycelia growth and spore germination of F. solani provides an inside to the likely behaviour of this devastating soilborne plant pathogen in nature and has important practical implications.

Asunto(s)

Fusarium/crecimiento & desarrollo , Presión Osmótica , Agua/metabolismo , Arachis/microbiología , Fusarium/efectos de los fármacos , Fusarium/efectos de la radiación , Glicerol/metabolismo , Enfermedades de las Plantas/microbiología , Polietilenglicoles/metabolismo , Microbiología del Suelo , Cloruro de Sodio/metabolismo , Temperatura

RESUMEN

Studies were conducted to determine the effect of osmotic and matric stress on germination and growth of two Fusarium solani strains, the etiological agent responsible of peanut brown root rot. Both strains had similar osmotic and matric potential ranges that allowed growth, being the latter one narrower. F. solani showed the ability to grow down to -14 MPa at 25 °C in non-ionic modified osmotic medium, while under matric stress this was limited to -8.4 MPa at 25 °C. However, both strains were seen to respond differently to decreasing osmotic and matric potentials, during early stages of germination. One strain (RC 338) showed to be more sensitive to matric than osmotic (non ionic) and the other one (RC 386) showed to be more sensitive to osmotic than matric imposed water stress. After 24 h of incubation, both isolates behaved similarly. The minimum water potential for germination was -8.4 MPa on glycerol amended media and -5.6 MPa for NaCl and PEG amended media, respectively. The knowledge of the water potential range which allow mycelia growth and spore germination of F. solani provides an inside to the likely behaviour of this devastating soilborne plant pathogen in nature and has important practical implications.

Asunto(s)

Fusarium/crecimiento & desarrollo , Presión Osmótica , Agua/metabolismo , Arachis/microbiología , Fusarium/efectos de los fármacos , Fusarium/efectos de la radiación , Glicerol/metabolismo , Enfermedades de las Plantas/microbiología , Polietilenglicoles/metabolismo , Microbiología del Suelo , Cloruro de Sodio/metabolismo , Temperatura

RESUMEN

N,N-diethyl-meta-toluamide (DEET) is a widely used insect repellent due to its high efficacy. In this work, micellar systems based on poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) triblock copolymer were developed and studied for the purpose of controlling the release and cutaneous permeation of DEET, using concentrated solutions of the copolymer Pluronic F127 to form thermoreversible gels. The formulations presented thermoreversible gelation above 5°C and altered rheological behavior at 15 and 25°C. The presence of the drug drastically changed the sol-gel transition temperatures. The micrographs suggest that DEET induced the formation of anisotropic structures, and Maltese Crosses were observed. The formulation containing 10wt% DEET and 15wt% Pluronic F127 presented sustained drug release for up to 7h. DEET release profile followed the Higuchi kinetics model. There was a reduction of approximately 35% in the amount of DEET absorbed through the skin after 6h. About 62% of DEET from the formulation consisting of Pluronic F127 and DEET remain retained on the skin. The anisotropic structure may constitute a barrier to diffusion and thereby controlling the drug release effectively. These tests suggest that the tested samples exhibit safety profile greater than some commercially available products.

Asunto(s)

DEET/química , Repelentes de Insectos/química , Micelas , Polietilenglicoles/química , Glicoles de Propileno/química , Animales , DEET/metabolismo , Preparaciones de Acción Retardada/química , Técnicas In Vitro , Repelentes de Insectos/metabolismo , Ratones , Poloxámero/química , Polietilenglicoles/metabolismo , Glicoles de Propileno/metabolismo , Reología , Piel/metabolismo , Absorción Cutánea , Solubilidad