Long-term behavioral and neurochemical paradoxical alterations elicited following intranasal application of a chlorpyrifos formulation in mice.

The intranasal (IN) administration route represents a pathway for xenobiotics to reach the brain. The present study aimed to address the long-term consequences of IN administration of a chlorpyrifos (CPF) commercial formulation (fCPF) in mice. For this purpose, adult male CF-1 mice were intranasally administered with fCPF (10 mg/kg/day) three days a week, for 2 and 4 weeks, respectively. Behavioral and biochemical analyses were conducted 3-7, and 7.5 months after the last IN fCPF administration, respectively. Following a 6-month fCPF-free washout period, fur appearance and body injuries scores improved in the fCPF-treated groups. Notably, spatial learning and memory enhancement was observed 4 and 7 months after the last IN fCPF administration. Changes in oxidative stress markers and the activities of enzymes involved in cholinergic and glutamatergic pathways were observed in different brain areas from fCPF-treated mice, still after 7.5 months from fCPF application. Altogether, these neurochemical disturbances could be responsible for the described behavioral observations.

From waste to food and bioinsecticides: An innovative system integrating Tenebrio molitor bioconversion and pyrolysis bio-oil production.

To achieve a waste-free clean production, the present study aimed to valorize an underused agroindustrial byproduct (rice bran) by mealworms bioconversion and produce bio-oil from pyrolysis of insect excreta (frass) as bioinsecticide. To reach the first goal, the suitability of rice bran (RB) versus standard diet, wheat bran (WB), was examined by determining feed conversion, growth performance, and nutritional profile of T. molitor larvae. RB diet was an appropriate feed substrate for breeding mealworms, as evidenced by their high survival rates, optimal feed conversion parameters, and its capability to support the growth and life cycle of this insect. Besides, RB did not affect soluble larval protein content but modified crude fat content and fatty acid profile. In order to address the second aim, egested frass from RB and WB were subjected to pyrolysis to obtain bio-oils. The main compound was acetic acid (≈37%) followed by 1,6-anhydro-ß-d-glucopyranose (from 16 to 25%), as measured by GC-MS analysis. Nitrogen-containing chemicals accounted for ≈10%. Frass bio-oils could represent a novel source of bioinsecticides due to their bioeffectiveness in insect pests of economic importance (Plodia interpunctella and Tribolium castaneum) and medical interest (Culex pipiens pipiens). For P. interpunctella adults, frass bio-oils produced insecticidal activity by fumigant and contact exposure whereas for T. castaneum adults, just fumigant. By a miniaturized model that simulates semireal storage conditions, it was seen that, on T. castaneum, frass RB bio-oil generated higher repellent effect than frass WB. Finally, bio-oils proved to have larvicidal activity against Cx. p. pipiens.

Behavioral and neurochemical impairments after intranasal administration of chlorpyrifos formulation in mice.

Among the most relevant environmental factors associated with the etiology of neurodegenerative disorders are pesticides. Spray drift or volatilization generates pesticide dispersion after its application. In addition, inhalation or intranasal (IN) administration of xenobiotics constitutes a feasible route for substance delivery to the brain. This study investigates the behavioral and neurochemical effects of IN exposure to a commercial formulation of chlorpyrifos (fCPF). Adult male CF-1 mice were intranasally administered with fCPF (3-10 mg/kg/day) three days a week, for 2 weeks. Behavioral and biochemical analyses were conducted 20 and 30 days after the last IN fCPF administration, respectively. No significant behavioral or biochemical effects were observed in the 3 mg/kg fCPF IN exposure group. However, animals exposed to 10 mg/kg fCPF showed anxiogenic behavior and recognition memory impairment, with no effects on locomotor activity. In addition, the IN administration of 10 mg/kg fCPF altered the redox balance, modified the activity of enzymes belonging to the cholinergic and glutamatergic pathways, and affected glucose metabolism, and cholesterol levels in different brain areas. Taken together, these observations suggest that these biochemical imbalances could be responsible for the neurobehavioral disturbances observed after IN administration of fCPF in mice.

Rat developmental fluoride exposure affects retention memory, leads to a depressive-like behavior, and induces biochemical changes in offspring rat brains.

Water is the principal source of human exposure to fluoride (F). The high permeability of the placenta and blood-brain barrier to F during the intrauterine life up to the end of lactation may be crucial to neurological fetus development. Therefore, this study explores the effects of 5 and 10 mg/l F exposure during entire gestation and lactation periods, through neurobehavioral and biochemical tests performed on 90-day-old male offspring rats. The present study shows that pre and peri-natal exposure to F doses that are in the range of those found in groundwater sources in Argentina affects long-term memory and leads to a depressive-like behavior in 90-day-old male pup. Furthermore, the purpose of the investigation was to find out the possible biochemical changes through which the pre and peri-natal F-administration could generate such behavioral variations. We found alterations in transaminases, acetylcholinesterase, and alkaline phosphatase enzymes activity in specific brain areas (the prefrontal cortex, the striatum, and the hippocampus), together with findings regarding misbalanced oxidative stress. In conclusion, F exposure during the early stages of rat development alters brain-oxidative stress markers as well as the activity of enzymes implicated in cholinergic and glutamatergic systems. These molecular changes could contribute to the neurobehavioral alterations described in the present investigation.

Neurobehavioral and neurochemical effects in rats offspring co-exposed to arsenic and fluoride during development.

Arsenic (iAs) and fluoride (F) are ubiquitous in the environment. All over the world, in many countries, thousands of people are suffering from the toxic effects of arsenicals ad fluorides. These two elements are recognized worldwide as the most serious inorganic contaminants in drinking water. When two different types of toxicants are simultaneously going inside the human body they may function independently or can act as synergistic or antagonistic to one another. Although there have been reports in literature of individual toxicity of iAs and F, however, not much is known about the effects following the combined exposure to the toxicants above mentioned. In this work, we investigated the effect of the co-exposure to low levels of iAs/F through drinking water during pregnancy and lactation on central nervous system functionality in the exposed rats offspring. Wistar rats were exposed to one of these solutions: 0.05 mg/L iAs and 5 mg/L F (Concentration A) or 0.10 mg/L iAs and 10 mg/L F (Concentration B) from gestational day 0 up to post-gestational day 21. Sensory-motor reflexes a Functional Observational Battery and the locomotor activity in an open field were assessed in offspring. Additionally, the transaminases, acethylcholinesterase and catalase levels in the striatum were determined to elucidate the possible molecular mechanisms involved in locomotor and neurobehavioral disorders. The results showed that iAs/F exposition during development produces a delay reach the maturity of sensorimotor reflexes. A decrease in the nociceptive reflex response, and increase in the locomotor activity in adult rats offspring were observed. The increase in oxidative stress, the inhibition of transaminases enzymes and the inhibition of AChE in the striatum may partially regulate all the neurobehavioral disorders observed.

Magnetic nanotechnology for diclofenac remediation: molecular basis of drug adsorption and neurobehavioral toxicology as a preliminary study for safe application.

Diclofenac is a commercial non-steroidal anti-inflammatory drug commonly present as a pollutant in naturally occurring water sources and wastewaters. In this work, the adsorption of diclofenac onto chitosan-coated magnetic nanosystems is proposed as a possible tool for remediation. Experimental and theoretical studies have been carried out to reveal the mechanisms associated with diclofenac interactions among all the components of the nanosystem. Mechanisms are presented, analyzed and discussed. A toxicological study in mice was carried out to evaluate the parameters associated with neurotoxicity of the nanodevice. The elucidation of the mechanisms implied in the adsorption process of diclofenac onto magnetic chitosan nanocomposites suggests that diclofenac remediation from water is possible by adsorption onto chitosan. The strategy innovates the commonly used methodologies for diclofenac remediation from pharmaceutical wastes. This magnetic nanotechnology would not induce damage on the nervous system in a murine model, in case of traces remaining in water sources.

Intranasal glyphosate-based herbicide administration alters the redox balance and the cholinergic system in the mouse brain.

Pesticide exposure is associated with cognitive and psychomotor disorders. Glyphosate-based herbicides (GlyBH) are among the most used agrochemicals, and inhalation of GlyBH sprays may arise from frequent aerial pulverizations. Previously, we described that intranasal (IN) administration of GlyBH in mice decreases locomotor activity, increases anxiety, and impairs recognition memory. Then, the aim of the present study was to investigate the mechanisms involved in GlyBH neurotoxicity after IN administration. Adult male CF-1 mice were exposed to GlyBH IN administration (equivalent to 50 mg/kg/day of Gly acid, 3 days a week, during 4 weeks). Total thiol content and the activity of the enzymes catalase, acetylcholinesterase and transaminases were evaluated in different brain areas. In addition, markers of the cholinergic and the nigrostriatal pathways, as well as of astrocytes were evaluated by fluorescence microscopy in coronal brain sections. The brain areas chosen for analysis were those seen to be affected in our previous study. GlyBH IN administration impaired the redox balance of the brain and modified the activities of enzymes involved in cholinergic and glutamatergic pathways. Moreover, GlyBH treatment decreased the number of cholinergic neurons in the medial septum as well as the expression of the α7-acetylcholine receptor in the hippocampus. Also, the number of astrocytes increased in the anterior olfactory nucleus of the exposed mice. Taken together, these disturbances may contribute to the neurobehavioural impairments reported previously by us after IN GlyBH administration in mice.

Effects of Perinatal Fluoride Exposure on Short- and Long-Term Memory, Brain Antioxidant Status, and Glutamate Metabolism of Young Rat Pups.

Exposure to fluoride (F) during the development affects central nervous system of the offspring rats which results in the impairment of cognitive functions. However, the exact mechanisms of F neurotoxicity are not clearly defined. To investigate the effects of perinatal F exposure on memory ability of young rat offspring, dams were exposed to 5 and 10 mg/L F during gestation and lactation. Additionally, we evaluated the possible underlying neurotoxic mechanisms implicated. The results showed that the memory ability declined in 45-day-old offspring, together with a decrease of catalase and glutamate transaminases activity in specific brain areas. The present study reveals that exposure to F in early stages of rat development leads to impairment of memory in young offspring, highlighting the alterations of oxidative stress markers as well as the activity of enzymes involved in the glutamatergic system as a possible mechanisms of neurotoxicity.

Prenatal Exposure to Cadmium During Organogenesis Impairs Memory in Young Rats.

Pregnant rats were treated with 0.3 and 0.6 mg cadmium (CdCl2)/kg injected subcutaneously on a daily basis from gestational day 7 to day 15 (organogenesis period). One control group was not injected and other received saline. The 45-day-old offspring were tested in a step-down inhibitory avoidance to evaluate short-term and long-term memory and in a radial maze for the study of spatial memory. These studies showed that gestational exposure to 0.6 mg Cd/kg produced in the male offspring a significant impairment in the retention of long-term memory evaluated 24 hours after training in the step-down inhibitory avoidance. The radial maze also demonstrated that the male offspring prenatally exposed to 0.6 mg Cd presented a significant deficit in the retention of spatial memory evaluated 42 days after training. These results demonstrate that the exposure to Cd during organogenesis may affect the retention of some types of memory.

Alterations in the memory of rat offspring exposed to low levels of fluoride during gestation and lactation: Involvement of the α7 nicotinic receptor and oxidative stress.

Daily exposure to fluoride (F) depends mainly on the intake of this element with drinking water. When administered during gestation and lactation, F has been associated with cognitive deficits in the offspring. However, the mechanisms underlying the neurotoxicity of F remain obscure. In the current study, we investigated the effects of oral exposure to low levels of F during the gestational and lactation periods, on the memory of adult female rat offspring. We also considered a possible underlying neurotoxic mechanism. Our results showed that this exposure reduced step-down latency in the inhibitory avoidance task, and decreased both mRNA expression of the α7 nicotinic receptor (nAChR) and catalase activity in hippocampus. Our data indicates that low F concentrations administrated during gestation and lactation decrease the memory of 90-day-old female offspring. This suggests that the mechanism might be connected with an α7 nAChR deficit in the hippocampus, induced by oxidative stress.

Low arsenic concentrations impair memory in rat offpring exposed during pregnancy and lactation: Role of α7 nicotinic receptor, glutamate and oxidative stress.

Inorganic arsenic (iAs) is an important natural pollutant. Millions of individuals worldwide drink water with high levels of iAs. Arsenic exposure has been associated to cognitive deficits. However, the underlying mechanisms remain unknown. In the present work we investigated in female adult offspring the effect of the exposure to low arsenite sodium levels through drinking water during pregnancy and lactation on short- and long-term memory. We also considered a possible underlying neurotoxic mechanism. Pregnant rats were exposed during pregnancy and lactation to environmentally relevant iAs concentrations (0.05 and 0.10 mg/L). In 90-day-old female offspring, short-term memory (STM) and long-term memory (LTM) were evaluated using a step-down inhibitory avoidance task. In addition, we evaluated the α7 nicotinic receptor (α7-nAChR) expression, the transaminases and the oxidative stress levels in hippocampus. The results showed that the exposure to 0.10 mg/L iAs in this critical period produced a significant impairment in the LTM retention. This behavioral alteration might be associated with several events that occur in the hippocampus: decrease in α7-nAChR expression, an increase of glutamate levels that may produce excitotoxicity, and a decrease in the antioxidant enzyme catalase (CAT) activity.

Perinatal Glyphosate-Based Herbicide Exposure in Rats Alters Brain Antioxidant Status, Glutamate and Acetylcholine Metabolism and Affects Recognition Memory.

Glyphosate-based herbicides (Gly-BHs) lead the world pesticide market. Although are frequently promoted as safe and of low toxicity, several investigations question its innocuousness. Previously, we described that oral exposure of rats to a Gly-BH during pregnancy and lactation decreased locomotor activity and anxiety in the offspring. The aim of the present study was to evaluate the mechanisms of neurotoxicity of this herbicide. Pregnant Wistar rats were supplied orally with 0.2 and 0.4% of Gly-BH (corresponding to 0.65 and 1.30 g/l of pure Gly, respectively) from gestational day (GD) 0, until weaning (postnatal day, PND, 21). Oxidative stress markers were determined in whole brain homogenates of PND90 offspring. The activity of acetylcholinesterase (AChE), transaminases, and alkaline phosphatase (AP) were assessed in prefrontal cortex (PFC), striatum, and hippocampus. Recognition memory was evaluated by the novel object recognition test. Brain antioxidant status was altered in Gly-BH-exposed rats. Moreover, AChE and transaminases activities were decreased and AP activity was increased in PFC, striatum and hippocampus by Gly-BH treatment. In addition, the recognition memory after 24 h was impaired in adult offspring perinatally exposed to Gly-BH. The present study reveals that exposure to a Gly-BH during early stages of rat development affects brain oxidative stress markers as well as the activity of enzymes involved in the glutamatergic and cholinergic systems. These alterations could contribute to the neurobehavioral variations reported previously by us, and to the impairment in recognition memory described in the present work.

Magnetic nanoparticles for drug targeting: from design to insights into systemic toxicity. Preclinical evaluation of hematological, vascular and neurobehavioral toxicology.

A simple two-step drug encapsulation method was developed to obtain biocompatible magnetic nanocarriers for the potential targeted treatment of diverse diseases. The nanodevice consists of a magnetite core coated with chitosan (Chit@MNPs) as a platform for diclofenac (Dic) loading as a model drug (Dic-Chit@MNPs). Mechanistic and experimental conditions related to drug incorporation and quantification are further addressed. This multi-disciplinary study aims to elucidate the toxicological impact of the MNPs at hematological, vascular, neurological and behavioral levels. Blood compatibility assays revealed that MNPs did not affect either erythrosedimentation rates or erythrocyte integrity at the evaluated doses (1, 10 and 100 µg mL-1). A microscopic evaluation of blood smears indicated that MNPs did not induce morphological changes in blood cells. Platelet aggregation was not affected by MNPs either and just a slight diminution was observed with Dic-Chit@MNPs, an effect possibly due to diclofenac. The examined formulations did not exert cytotoxicity on rat aortic endothelial cells and no changes in cell viability or their capacity to synthesize NO were observed. Behavioral and functional nervous system parameters in a functional observational battery were assessed after a subacute treatment of mice with Chit@MNPs. The urine pools of the exposed group were decreased. Nephritis and an increased number of megakaryocytes in the spleen were observed in the histopathological studies. Sub-acute exposure to Chit@MNPs did not produce significant changes in the parameters used to evaluate neurobehavioral toxicity. The aspects focused on within this manuscript are relevant at the pre-clinical level providing new and novel knowledge concerning the biocompatibility of magnetic nanodevices for biomedical applications.

Influence of chitosan coating on magnetic nanoparticles in endothelial cells and acute tissue biodistribution.

Chitosan coating on magnetic nanoparticles (MNPs) was studied on biological systems as a first step toward the application in the biomedical field as drug-targeted nanosystems. Composition of MNPs consists of magnetite functionalized with oleic acid and coated with the biopolymer chitosan or glutaraldehyde-cross-linked chitosan. The influence of the biopolymeric coating has been evaluated by in vitro and in vivo assays on the effects of these MNPs on rat aortic endothelial cells (ECs) viability and on the random tissue distribution in mice. Results were correlated with the physicochemical properties of the nanoparticles. Nitric oxide (NO) production by ECs was determined, considering that endothelial NO represents one of the major markers of ECs function. Cell viability was studied by MTT assay. Different doses of the MNPs (1, 10 and 100 µg/mL) were assayed, revealing that MNPs coated with non-cross-linked chitosan for 6 and 24 h did not affect neither NO production nor cell viability. However, a significant decrease in cell viability was observed after 36 h treatment with the highest dose of this nanocarrier. It was also revealed that the presence and dose of glutaraldehyde in the MNPs structureimpact on the cytotoxicity. The study of the acute tissue distribution was performed acutely in mice after 24 h of an intraperitoneal injection of the MNPs and sub acutely, after 28 days of weekly administration. Both formulations greatly avoided the initial clearance by the reticuloendothelial system (RES) in liver. Biological properties found for N1 and N2 in the performed assays reveal that chitosan coating improves biocompatibility of MNPs turning these magnetic nanosystems as promising devices for targeted drug delivery.

Exposure to a glyphosate-based herbicide during pregnancy and lactation induces neurobehavioral alterations in rat offspring.

The impact of sub-lethal doses of herbicides on human health and the environment is a matter of controversy. Due to the fact that evidence particularly of the effects of glyphosate on the central nervous system of rat offspring by in utero exposure is scarce, the purpose of the present study was to assess the neurobehavioral effects of chronic exposure to a glyphosate-containing herbicide during pregnancy and lactation. To this end, pregnant Wistar rats were exposed through drinking water to 0.2% or 0.4% of a commercial formulation of glyphosate (corresponding to a concentration of 0.65 or 1.30g/L of glyphosate, respectively) during pregnancy and lactation and neurobehavioral alterations in offspring were analyzed. The postnatal day on which each pup acquired neonatal reflexes (righting, cliff aversion and negative geotaxis) and that on which eyes and auditory canals were fully opened were recorded for the assessment of sensorimotor development. Locomotor activity and anxiety levels were monitored via open field test and plus maze test, respectively, in 45- and 90-day-old offspring. Pups exposed to a glyphosate-based herbicide showed early onset of cliff aversion reflex and early auditory canal opening. A decrease in locomotor activity and in anxiety levels was also observed in the groups exposed to a glyphosate-containing herbicide. Findings from the present study reveal that early exposure to a glyphosate-based herbicide affects the central nervous system in rat offspring probably by altering mechanisms or neurotransmitter systems that regulate locomotor activity and anxiety.

Neurobehavioural effects of exposure to fluoride in the earliest stages of rat development.

It is known that exposure to high concentrations of Fluoride (F) produces deleterious health effects in human population. However, in the last years it has been concluded that low concentrations of F may have adverse health effects as well. Transplacental passage of F and its incorporation into foetal tissues has been demonstrated. Therefore, the purpose of the present work was to study the effects of the exposure to low levels of F during pregnancy and lactation on the central nervous system functionality. Wistar rats were exposed to low F concentrations (5 and 10 mg/l) during pregnancy and lactation. Sensorimotor reflexes in the each pup were analysed and the postnatal day on which both eyes and auditory canals were opened was recorded. Locomotor activity and anxiety were subsequently analysed in 45- and 90-day-old offspring by an open field test and plus maze test, respectively. A significant delay in the development of eye opening was observed in all offspring whose mothers had been exposed to the two F concentrations tested. Exposure to 5 and 10 mg/l F was also found to significantly decrease locomotor activity only in 90-day-old male and female offspring. A low index of anxiety in the young females and in all adult offspring exposed to the two F concentrations tested was also detected. Taken together, findings from the present study show that exposure to low F concentrations during pregnancy and lactation produces dysfunction in the central nervous system mechanisms which regulate motor and sensitive development, locomotor activity and anxiety

Locomotor activity and sensory-motor developmental alterations in rat offspring exposed to arsenic prenatally and via lactation.

Arsenic (As) is one of the most toxic naturally occurring contaminants in the environment. The major source of human exposure to inorganic As (iAs) is through contaminated drinking water. Although both genotoxicity and carcinogenicity derived from this metalloid have been thoroughly studied, the effects of iAs on the development and function of the central nervous system (CNS) have received less attention and only a few studies have focused on neurobehavioral effects. Thus, in order to characterize developmental and behavioral alterations induced by iAs exposure, pregnant Wistar rats were exposed to 0.05 and 0.10 mg/L iAs through drinking water during gestation and lactation. Sensory-motor reflexes in each pup were analyzed and the postnatal day when righting reflex, cliff aversion and negative geotaxis were recorded. Functional Observational Battery (FOB) and locomotor activity in an open field were assessed in 90-day-old offspring. Results show that rats exposed to low iAs concentrations through drinking water during early development evidence a delay in the development of sensory-motor reflexes. Both FOB procedure and open-field tests showed a decrease in locomotor activity in adult rats. This study reveals that exposure to the above-mentioned iAs concentrations produces dysfunction in the CNS mechanisms whose role is to regulate motor and sensory development and locomotor activity.

Enhanced analgesic properties and reduced ulcerogenic effect of a mononuclear copper(II) complex with fenoprofen in comparison to the parent drug: promising insights in the treatment of chronic inflammatory diseases.

Analgesic and ulcerogenic properties have been studied for the copper(II) coordination complex of the nonsteroidal anti-inflammatory drug Fenoprofen and imidazole [Cu(fen)2(im)2] (Cu: copper(II) ion; fen: fenoprofenate anion from Fenoprofen, im: imidazole). A therapeutic dose of 28 mg/kg was tested for [Cu(fen)2(im)2] and 21 mg/kg was employed for Fenoprofen calcium, administered by oral gavage in female mice to compare the therapeutic properties of the new entity. The acetic acid induced writhing test was employed to study visceral pain. The percentage of inhibition in writhing and stretching was 78.9% and 46.2% for the [Cu(fen)2(im)2] and Fenoprofen calcium, respectively. This result indicates that the complex could be more effective in diminishing visceral pain. The formalin test was evaluated to study the impact of the drugs over nociceptive and inflammatory pain. The complex is a more potent analgesic on inflammatory pain than the parent drug. Ulcerogenic effects were evaluated using a model of gastric lesions induced by hypothermic-restraint stress. Fenoprofen calcium salt caused an ulcer index of about 79 mm(2) while the one caused by [Cu(fen)2(im)2] was 22 mm(2). The complex diminished the development of gastric mucosal ulcers in comparison to the uncomplexed drug. Possible mechanisms of action related to both therapeutic properties have been discussed.

Anti-nociceptive activity and toxicity evaluation of Cu(II)-fenoprofenate complexes in mice.

The proposed curative properties of copper(II)-non-steroidal anti-inflammatory drugs (NSAIDs) have led to the development of numerous copper(II)-NSAID complexes with enhanced anti-inflammatory activity. In this work, the antinociceptive and toxic effects of two new coordination complexes: Cu2(fen)4(caf)2 [fen: fenoprofenate anion; caf: caffeine] and Cu2(fen)4(dmf)2 [dmf: N-N'-dimethylformamide] were evaluated in mice. The antinociceptive effect was evaluated with two models: acetic acid-induced writhing response and formalin test. For the sub-acute exposure, the complexes were added to the diet at different doses for 28days. Behavioral and functional nervous system parameters in a functional observational battery were assessed. Also, hematological, biochemical and histopathological studies were performed. Cu2(fen)4(caf)2 and Cu2(fen)4(dmf)2 significantly decreased the acetic acid-induced writhing response and the licking time on the late phase in the formalin test with respect to the control and fenoprofen salt groups. The sub-acute exposure to Cu2(fen)4(caf)2 complex increased the motor activity, the number of rearings and the arousal with respect to the control and fenoprofen salt groups. These impaired parameters in mice exposed to Cu2(fen)4(caf)2 can be attributable to the presence of caffeine as stimulating agent. On the other hand, all exposed groups decreased the urine pools in the functional observational battery and increased the plasmatic urea. These effects could be due to the decrease in the glomerular filtration caused by NSAIDs. In conclusion, both complexes Cu2(fen)4(dmf)2 and Cu2(fen)4(caf)2 were more potent antinociceptive agents than fenoprofen salt. Sub-acute exposure to different doses of these complexes did not produce significant changes in the parameters that evaluate toxicity.