HR/MS-based lipidome analysis of rat brain modulated by tolcapone.

Lipids play key roles in the body, influencing cellular regulation, function, and signalling. Tolcapone, a potent catechol-O-methyltransferase (COMT) inhibitor described to enhance cognitive performance in healthy subjects, was previously shown to impact fatty acid ß-oxidation and oxidative phosphorylation. However, its impact on the brain lipidome remains unexplored. Hence, this study aimed to assess how tolcapone affects the lipidome of the rat pre-frontal cortex (PFC), a region of the brain highly relevant to tolcapone therapeutic effect, while evaluating its influence on operant behaviour. Tolcapone at 20 mg/kg was chronically administered to Wistar rats during a behavioural task and an untargeted liquid chromatography high-resolution mass spectrometry (LC-HR/MS) approach was employed to profile lipid species. The untargeted analysis identified 7227 features, of which only 33% underwent statistical analysis following data pre-processing. The results revealed an improved cognitive performance and a lipidome remodelling promoted by tolcapone. The lipidomic analysis showed 32 differentially expressed lipid species in tolcapone-treated animals (FC ≥ 1.2, p-value ≤ 0.1), and among these several triacylglycerols, cardiolipins and N-acylethanolamine (NAE 16:2) were found upregulated whereas fatty acids, hexosylceramides, and several phospholipids including phosphatidylcholines and phosphatidylethanolamines were downregulated. These preliminary findings shed light on tolcapone impact on lipid pathways within the brain. Although tolcapone improved cognitive performance and literature suggests the significance of lipids in cognition, this study did not conclusively establish that lipids directly drove or contributed to this outcome. Nevertheless, it underscores the importance of lipid modulation and encourages further exploration of tolcapone-associated mechanisms in the central nervous system (CNS).

Predictors of psychotherapy dropout in patients with borderline personality disorder: A systematic review.

INTRODUCTION: Borderline personality disorder (BPD) is a highly debilitating psychiatric condition. Despite the expansion of new BPD specific forms of psychotherapy in the last few decades, high dropout rates have been reported in these treatments. Treatment discontinuation is associated with poor patient outcomes, inefficient resource utilization and the demoralization of healthcare providers. METHODS: In order to identify predictors of psychotherapy dropout among patients with BPD, a systematic search of Medline, the Cochrane Library, PsycInfo and PsycArticles was conducted. Studies included were randomized-controlled trials in which patients diagnosed with BPD were exposed to a therapeutic intervention consisted of an evidence-based psychotherapy. The quality of evidence in the studies was assessed through the use of revised Cochrane risk of bias tool. RESULTS: Six articles, incorporating four types of psychotherapy programmes, were included. Overall, the studies present low risk of attrition and reporting bias and unclear risk of selection, performance and detection bias. Patients with weaker therapeutic alliance scores and higher hostility presented with higher dropout rates. In contrast, better mindfulness skills and greater performance in specific neuropsychological domains, such as memory and executive control, were identified as predictive of lower risk of dropout. Sociodemographic variables and treatment history did not influence treatment retention. CONCLUSIONS: Factors that influence discontinuation should be taken into consideration in future treatment programmes, in an effort to optimize retention. Qualitative assessments of patients' reasons for dropping out may also help guide adjustments.

Exploring the response of a key Mediterranean gorgonian to heat stress across biological and spatial scales.

Understanding the factors and processes that shape intra-specific sensitivity to heat stress is fundamental to better predicting the vulnerability of benthic species to climate change. Here, we investigate the response of a habitat-forming Mediterranean octocoral, the red gorgonian Paramuricea clavata (Risso, 1826) to thermal stress at multiple biological and geographical scales. Samples from eleven P. clavata populations inhabiting four localities separated by hundreds to more than 1500 km of coast and with contrasting thermal histories were exposed to a critical temperature threshold (25 °C) in a common garden experiment in aquaria. Ten of the 11 populations lacked thermotolerance to the experimental conditions provided (25 days at 25 °C), with 100% or almost 100% colony mortality by the end of the experiment. Furthermore, we found no significant association between local average thermal regimes nor recent thermal history (i.e., local water temperatures in the 3 months prior to the experiment) and population thermotolerance. Overall, our results suggest that local adaptation and/or acclimation to warmer conditions have a limited role in the response of P. clavata to thermal stress. The study also confirms the sensitivity of this species to warm temperatures across its distributional range and questions its adaptive capacity under ocean warming conditions. However, important inter-individual variation in thermotolerance was found within populations, particularly those exposed to the most severe prior marine heatwaves. These observations suggest that P. clavata could harbor adaptive potential to future warming acting on standing genetic variation (i.e., divergent selection) and/or environmentally-induced phenotypic variation (i.e., intra- and/or intergenerational plasticity).

Lactate modulates cardiac gene expression in mice during acute physical exercise.

The aim of the present study was to verify the role of lactate as a signaling molecule in cardiac tissue under physiological conditions. C57BL6/J male mice were submitted to acute running bouts on a treadmill at different exercise intensities (30, 60, and 90% of maximal speed - Smax) under the effect of two doses (0.5 and 5 mM) of α-cyano-4-hydroxycynnamate (CINN), a blocker of lactate transporters. Cardiac lactate levels, activity of the enzymes of glycolytic [hexokinase (HK) and lactate dehydrogenase (LDH)] and oxidative metabolism [citrate synthase (CS)], and expression of genes also related to metabolism [LDH, nuclear factor erythroid 2-related factor 2 (NRF-2), cytochrome oxidase IV (COX-IV), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α)] were evaluated. Elevated cardiac lactate levels were observed after high intensity running at 90% of Smax, which were parallel to increased activity of the HK and CS enzymes and mRNA levels of PGC-1α and COX-IV. No changes were observed in cardiac lactate levels in mice running at lower exercise intensities. Interestingly, prior intraperitoneal administration (15 min) of CINN (0.5 mM) significantly reduced cardiac lactate concentration, activities of HK and CS, and mRNA levels of PGC-1α and COX-IV in mice that ran at 90% of Smax. In addition, cardiac lactate levels were significantly correlated to both PGC-1α and COX-IV cardiac gene expression. The present study provides evidence that cardiac lactate levels are associated to gene transcription during an acute bout of high intensity running exercise.

Lactate modulates cardiac gene expression in mice during acute physical exercise

The aim of the present study was to verify the role of lactate as a signaling molecule in cardiac tissue under physiological conditions. C57BL6/J male mice were submitted to acute running bouts on a treadmill at different exercise intensities (30, 60, and 90% of maximal speed - Smax) under the effect of two doses (0.5 and 5 mM) of α-cyano-4-hydroxycynnamate (CINN), a blocker of lactate transporters. Cardiac lactate levels, activity of the enzymes of glycolytic [hexokinase (HK) and lactate dehydrogenase (LDH)] and oxidative metabolism [citrate synthase (CS)], and expression of genes also related to metabolism [LDH, nuclear factor erythroid 2-related factor 2 (NRF-2), cytochrome oxidase IV (COX-IV), and peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC-1α)] were evaluated. Elevated cardiac lactate levels were observed after high intensity running at 90% of Smax, which were parallel to increased activity of the HK and CS enzymes and mRNA levels of PGC-1α and COX-IV. No changes were observed in cardiac lactate levels in mice running at lower exercise intensities. Interestingly, prior intraperitoneal administration (15 min) of CINN (0.5 mM) significantly reduced cardiac lactate concentration, activities of HK and CS, and mRNA levels of PGC-1α and COX-IV in mice that ran at 90% of Smax. In addition, cardiac lactate levels were significantly correlated to both PGC-1α and COX-IV cardiac gene expression. The present study provides evidence that cardiac lactate levels are associated to gene transcription during an acute bout of high intensity running exercise.

Receptor-mediated PLGA nanoparticles for glioblastoma multiforme treatment.

Glioblastoma multiforme is the most lethal type of brain tumor and the established therapy only extends patients survival to approximately one year. Its first-line treatment is based on of chemotherapy with the alkylating agent temozolomide (TMZ). As many other chemotherapeutic drugs, TMZ presents several limitations as high toxicity and low bioavailability. The delivery of TMZ using poly(lactic-co-glycolic acid) nanoparticles is proposed in this work. Stable nanoparticles functionalized with a OX26 type monoclonal antibody for transferrin receptor were developed, targeting the glioblastoma tumor cells, since these cells are known for overexpressing this receptor. The release profile of TMZ from the nanoparticles was studied mimicking physiological conditions, and targeted cellular internalization was also investigated. Two glioblastoma cell lines - U215 and U87 - were used to evaluate the in vitro cytotoxicity of the drug, showing that the prepared nanocarriers enhance the anticancer activity of TMZ. The functionalization with the monoclonal antibody for transferrin receptor proved to be advantageous in enhancing the cellular internalization in glioblastoma cells.

Homer2 within the central nucleus of the amygdala modulates withdrawal-induced anxiety in a mouse model of binge-drinking.

A history of binge-drinking decreases protein expression of the glutamate-related scaffolding protein Homer2 within the central nucleus of the amygdala (CEA), coinciding with behavioral signs of negative affect. To assess the functional relevance of this protein change for withdrawal-induced hyper-anxiety, adult (PND 56) and adolescent (PND 28) male C57BL/6J mice were administered an intra-CEA infusion of an adeno-associated viral vector (AAV) carrying either cDNA to express Homer2 (H2-cDNA) or GFP as control. Mice underwent 14 days of binge-drinking under multi-bottle, limited-access conditions and were assayed for behavioral signs of negative affect during withdrawal using the light-dark box, marble burying, and forced swim tests (FST). Following behavioral testing, all animals experienced 5 days of drinking to evaluate the effects of prior alcohol experience and Homer2 manipulation on subsequent alcohol consumption. During protracted (4 weeks) withdrawal, adolescent alcohol-experienced GFP controls showed increased signs of negative affect across all 3 assays, compared to water-drinking GFP animals, and also showed elevated alcohol consumption during the subsequent drinking period. Homer2-cDNA infusion in adolescent-onset alcohol-drinking animals was anxiolytic and reduced subsequent alcohol consumption. Conversely, Homer2-cDNA was anxiogenic and increased drinking in water-drinking adolescents. Unfortunately, the data from adult-onset alcohol-drinking animals were confounded by low alcohol consumption and negligible behavioral signs of anxiety. Nevertheless, the present results provide novel cause-effect evidence supporting a role for CEA Homer2 in the regulation of both basal anxiety and the time-dependent intensification of negative affective states in individuals with a history of binge-drinking during adolescence.

Variability in prescription opioid intake and reinforcement amongst 129 substrains.

Opioid abuse in the United States has reached epidemic proportions, with treatment admissions and deaths associated with prescription opioid abuse quadrupling over the past 10 years. Although genetics are theorized to contribute substantially to inter-individual variability in the development, severity and treatment outcomes of opioid abuse/addiction, little direct preclinical study has focused on the behavioral genetics of prescription opioid reinforcement and drug-taking. Herein, we employed different 129 substrains of mice currently available from The Jackson Laboratory (129S1/SvlmJ, 129X1/SvJ, 129S4/SvJaeJ and 129P3/J) as a model system of genetic variation and assayed mice for oral opioid intake and reinforcement, as well as behavioral and somatic signs of dependence. All substrains exhibited a dose-dependent increase in oral oxycodone and heroin preference and intake under limited-access procedures and all, but 129S1/SvlmJ mice, exhibited oxycodone reinforcement. Relative to the other substrains, 129P3/J mice exhibited higher heroin and oxycodone intake. While 129X1/SvJ exhibited the highest anxiety-like behavior during natural opioid withdrawal, somatic and behavior signs of precipitated withdrawal were most robust in 129P3/J mice. These results demonstrate the feasibility and relative sensitivity of our oral opioid self-administration procedures for detecting substrain differences in drug reinforcement/intake among 129 mice, of relevance to the identification of genetic variants contributing to high vs. low oxycodone reinforcement and intake.

Adaptability and phenotypic stability of soybean cultivars for grain yield and oil content.

The aim of this study was to verify the adaptability and stability of soybean cultivars with regards to yield and oil content. Data of soybean yield and oil content were used from experiments set up in six environments in the 2011/12 and 2012/13 crop seasons in the municipalities of Patos de Minas, Uberaba, Lavras, and São Gotardo, Minas Gerais, Brazil, testing 36 commercial soybean cultivars of both conventional and transgenic varieties. The Wricke method and GGE biplot analysis were used to evaluate adaptability and stability of these cultivars. Large variations were observed in grain yield in relation to the different environments studied, showing that these materials are adaptable. The cultivars exhibited significant differences in oil content. The cultivars BRSGO204 (Goiânia) and BRSMG (Garantia) exhibited the greatest average grain yield in the different environments studied, and the cultivar BRSMG 760 SRR had the greatest oil content among the cultivars evaluated. Ecovalence was adopted to identify the most stable cultivars, and the estimates were nearly uniform both for grain yield and oil content, showing a variation of 0.07 and 0.01%, respectively. The GGE biplot was efficient at identifying cultivars with high adaptability and phenotype stability.

Catechol-O-methyltransferase activity in erythrocytes from patients with eating disorders.

PURPOSE: Abnormal feeding has been linked to disruptions in brain dopaminergic activity and recent studies have assessed the role of catechol-O-methyltransferase (COMT) in eating disorders. This is the first study to quantify the soluble catechol-O-methyltransferase (S-COMT) activity in erythrocytes from patients with anorexia nervosa (AN), bulimia nervosa (BN) and binge-eating disorder (BED) and the first study at all to evaluate the COMT on patients with BED. METHODS: Forty blood samples from patients with AN, BN and BED and healthy controls were drawn to evaluate S-COMT activity in erythrocytes by high-performance liquid chromatography and mass spectrometry. Since several patients were being treated with fluoxetine 20 mg, they were included in a different group (BN MED and BED MED). Liver homogenates from rats were used to evaluate baseline S-COMT activity in the presence of fluoxetine by the same in vitro procedures and assays. RESULTS: Erythrocyte S-COMT activity (pmol/mg prt/h) was significantly increased in patients with BN and BED (41.3 ± 6.8 and 41.4 ± 14, respectively) compared to control group (25.3 ± 9.7). In fluoxetine-treated patients with BN, S-COMT activity (15.9 ± 8.8) was decreased compared to the other BN group; however, in BED group, the difference between BED MED and BED was not observed. In patients with AN, no significant difference was found compared to controls. CONCLUSION: Patients with BN and BED presented higher S-COMT activity in erythrocytes, which is in agreement with previous studies on the literature addressing the high-activity COMT allele, Val158, as risk factor for eating disorders. Although in fluoxetine-treated patients with BN the activity of S-COMT was similar to the controls, this is not explained by a direct interaction between fluoxetine and S-COMT as verified in in vitro assays.

Doctors' awareness concerning primary immunodeficiencies in Brazil.

BACKGROUND: PIDs are a heterogeneous group of genetic illnesses, and delay in their diagnosis is thought to be caused by a lack of awareness among physicians concerning PIDs. The latter is what we aimed to evaluate in Brazil. METHODS: Physicians working at general hospitals all over the country were asked to complete a 14-item questionnaire. One of the questions described 25 clinical situations that could be associated with PIDs and a score was created based on percentages of appropriate answers. RESULTS: A total of 4026 physicians participated in the study: 1628 paediatricians (40.4%), 1436 clinicians (35.7%), and 962 surgeons (23.9%). About 67% of the physicians had learned about PIDs in medical school or residency training, 84.6% evaluated patients who frequently took antibiotics, but only 40.3% of them participated in the immunological evaluation of these patients. Seventy-seven percent of the participating physicians were not familiar with the warning signs for PIDs. The mean score of correct answers for the 25 clinical situations was 48.08% (±16.06). Only 18.3% of the paediatricians, 7.4% of the clinicians, and 5.8% of the surgeons answered at least 2/3 of these situations appropriately. CONCLUSIONS: There is a lack of medical awareness concerning PIDs, even among paediatricians, who have been targeted with PID educational programmes in recent years in Brazil. An increase in awareness with regard to these disorders within the medical community is an important step towards improving recognition and treatment of PIDs.

Urinary profile of catecholamines and metabolites in Parkinson patients with deep brain stimulation.

BACKGROUND AND PURPOSE: Deep brain stimulation of the subthalamic nucleus (DBS-STN) is thought to continuously alter the activity of STN neurons in Parkinson's disease (PD). A chronic decrease in the levodopa dose with continuous STN stimulation may induce plastic neuronal changes. OBJECTIVE: The objective of this work was to study urinary excretion of catecholamines in patients with PD before and after DBS-STN. METHODS: Twenty-three patients were submitted to DBS-STN, and evaluated before and after surgery with respect to catecholamines and metabolites in 24-h urine measured by high-performance liquid chromatography with electrochemical detection. RESULTS: Of the 23 patients evaluated, a significant decrease of about 60% in the urinary excretion of L-3,4-dihydroxyphenylalanine (L-DOPA; in nmol/mg creatinine/24 h) was observed 1 week after DBS-STN. Moreover, in 17 patients with a follow-up of 8 weeks after surgery, there was a further 50% decrease in urinary L-DOPA levels, dropping to about 75% of the values before surgery. There was also a significant decrease in dopamine (DA) and 3,4-dihydroxyphenylacetic acid (DOPAC) levels 1 week after DBS-STN that was no longer present 8 weeks after. A significant increase in the DA/l-DOPA ratio was observed 1 week after surgery, with a further increase 8 weeks after surgery. CONCLUSION: After DBS-STN, the DA/l-DOPA ratio, an indirect measure of DA synthesis, increased. These results show that DBS-STN may improve the efficacy of oral levodopa.

Anaphylaxis in Latin American children and adolescents: the Online Latin American Survey on Anaphylaxis (OLASA).

OBJECTIVES: To identify the main clinical manifestations, triggers, and treatment of severe allergic reactions (SAR) in children and adolescents (n=191, up to 18 years of age) seen by allergologists and registered in the Online Latin American Survey of Anaphylaxis (OLASA). RESULTS: 53.0% of the patients were males and the aetiological agent was identified in 85.5% of them as follows: foods (36.1%), drugs (27.7%), and insect stings (26.2%). The most common symptoms during an acute episode were cutaneous (94.2%), and respiratory (78.5%). Most patients were treated in emergency setting, yet only 34.6% received parenteral epinephrine and 14.3% had to be hospitalised. CONCLUSION: Cutaneous symptoms ranked the order of clinical presentation of SAR. Food was the main triggering agent in the younger cases and insect sting and drugs in the adolescents. Treatment provided for SAR was not appropriate. It is necessary to improve educational programmes in order to enhance the knowledge on this potentially fatal emergency.

Development of an amperometric biosensor for phenol detection.

With the aim of searching for an in situ method for monitoring phenol, Agaricus bisporus tissue with tyrosine activity was used as a biocomponent and an oxygen electrode used as a transducer to develop a biosensor. The experimental methodology investigated the relation between dissolved oxygen and phenol concentration using a standard solution. Biosensor calibration was evaluated by studying reaction time and tissue amount necessary to promote a reliable response and to minimize errors. The influence of air saturation of the sample and washing of the electrode was also investigated. Results showed that 5 g of mushroom tissue with a 1 min reaction time promoted the best biosensor response within a phenol concentration range of 5-10 ppm. Washing of the electrode did not change the performance of the analysis; however, initial air saturation caused less variation amongst the samples.

Risk of malnutrition among Brazilian institutionalized elderly: a study with the Mini Nutritional Assessment (MNA) questionnaire.

BACKGROUND: Brazilian population has passed for a process of demographic transition throughout latest years, characterized for the increase of the elderly population. Malnutrition is a serious problem to frail elderly. OBJECTIVE: The objective of this study was o evaluate the risk of malnutrition among institutionalized elderly resident in municipal shelters in the city of Rio de Janeiro, Brazil, using the tool Mini Nutritional Assessment (MNA). DESIGN: 344 institutionalized elderly aged over 60 years old were tested in a cross-sectional study using MNA. This tool classifies the nutricional status of the elderly in three groups: malnutrition (score < 17), risk of malnutrition (score 17 - 23,5) and well-nourished (score > = 24). Anthropometric measurements such as calf circumference (CC), mid-arm circumference (MAC) and Body mass index (BMI) were also evaluated. The variables were evaluated using the chi-square or ANOVA test. To correlate it was used Pearson's Correlation Coefficient (r). RESULTS: Mean age were 75.4 (+- 9.4) years old. Most of the elderly were female gender (59.6%). According to MNA 8.3% were with malnutrition, 55.6% at risk of malnutrition and 36.1% well-nourished. BMI classified 10.0% of the elderly as underweight. CC classified 10.0 % of them as inadequate in muscular mass. MNA was well correlated to BMI (r=0.412 p=0.000), age (r=-0.124 p=0.031), CC (r=0.399 p = 0.000) and MAC (r=0.391 p=0.000). CONCLUSION: Risk of malnutrition was high among the institutionalized elderly from public shelters in Rio de Janeiro - Brazil. MNA is a useful diagnostic tool for the identification on the frail elderly at risk of malnutrition.

Avaliação da atividade antimicrobiana in vitro do extrato de Tabernaemontana catharinensis A. DC / Evaluation of in vitro antimicrobial activity of Tabernaemontana catharinensis A. DC. extract

Diversas espécies de Tabernaemontana têm sido estudadas devido a diversidade de alcalóides com atividade farmacológica. O objetivo desse trabalho foi avaliar a capacidade antimicrobiana in vitro do extrato das cascas do caule de Tabernaemontana catharinensis A. DC.em cepas de Staphylococcus aureus e Pseudomonas aeruginosa, microrganismos causadores de diversas infecções. Os testes de susceptibilidade bacteriana foram realizados usando o método de Kirby Bauer, consistindo na difusão em disco do antibiótico em meio de cultivo Mueller Hinton. Os testes de inibição foram realizados com soluções do extrato bruto seco de T. catharinensis dissolvido em etanol 70 por cento (v/v) na concentração 1,0 mg mL-1, que aplicada nos discos de área 20 mm², apresentaram concentração de 0,005 mg mm-2. Como controle negativo, realizou-se ensaios com placas contendo P. aeruginosa, e discos com etanol 70 por cento (v/v), e como controle positivo, discos com os antibióticos ceftriaxona sódica (0,25 mg mm-2 de área do disco), tetraciclina (0,005 mg mm-2) e cefalexina (0,005 mg mm-2). A solução do extrato na concentração de 0,005 mg mm-2 inibiu o Staphylococcus aureus, com diâmetro médio do halo de 0,6 cm. O halo de inibição para o Pseudomonas aeruginosa foi em média 1,2 cm. A tetraciclina, a cefalexina, e o controle negativo (etanol 70 por cento v/v) não demonstraram ação antimicrobiana. O halo de inibição usando ceftriaxona foi em média 2,2 cm para P. aeruginosa e 1,0 cm para Staphylococcus aureus.

Several Tabernaemontana species have been studied due to their several alkaloids with pharmacological activity. The aim of this work was to evaluate the in vitro antimicrobial action of the extract from stem barks of Tabernaemontana catharinensis A. DC. against strains of Staphylococcus aureus and Pseudomonas aeruginosa, microorganisms that cause several infections. Bacterial susceptibility tests were performed by the Kirby-Bauer method, consisting in antibiotic disk diffusion in Mueller Hinton medium. Inhibition tests were performed with solutions of T. catharinensis dry crude extract dissolved in ethanol 70 percent (v/v) at 1.0 mg mL-1, which became 0.005 mg mm-2 when applied to 20 mm² disks. As negative control, assays were carried out in plates containing P. aeruginosa and disks with ethanol 70 percent (v/v). Positive control consisted of disks containing the antibiotics ceftriaxone sodium (0.25 mg mm-2 disk area), tetracycline (0.005 mg mm-2) and cephalexin (0.005 mg mm-2). Extract solution at 0.005 mg mm-2 inhibited Staphylococcus aureus, with 0.6cm halo mean diameter. The inhibition halo for Pseudomonas aeruginosa was on average 1.2 cm. Tetracycline, cephalexin and negative control (ethanol 70 percent v/v) did not show antimicrobial action, whereas ceftriaxone sodium resulted in 2.2 and 1.0cm mean inhibition halo diameters for P. aeruginosa and Staphylococcus aureus, respectively.

Agaricus bisporus as a source of tyrosinase for phenol detection for future biosensor development.

Phenols are toxic compounds that are present in several industrial wastewaters, so their quantification has great environmental importance. In order to permit an analytical methodology for in situ monitoring, this work aims to study the application of Agaricus bisporus tissue as a source of tyrosinase and the optimum reaction conditions for the development of a phenol biosensor. Such an enzyme is a polyphenol oxidase that transforms many different phenolic compounds into quinones. Experiments with fungi tissue were performed to evaluate different sizes of tissue (0.5, 1.0 and 1.5 cm), different temperatures (23.5 degrees C to 60 degrees C), and different pH values (6, 7 and 8) to quantify analytically phenol content. Amongst the tested conditions, those that had presented larger efficiency in phenol oxidation were attained with the fungal tissue size of 1 cm, at pH 8.0, in the temperature range from 35 degrees C to 45 degrees C.

Effect of (-)-Delta(9)-tetrahydrocannabinoid on the hepatic redox state of mice.

(-)-Delta(9)-Tetrahydrocannabinol (Delta(9)-THC), a psychoactive component of marijuana, has been reported to induce oxidative damage in vivo and in vitro. In this study, we administered Delta(9)-THC to healthy C57BL/6J mice aged 15 weeks in order to determine its effect on hepatic redox state. Mice were divided into 3 groups: Delta(9)-THC (N = 10), treated with 10 mg/kg body weight Delta(9)-THC daily; VCtrl (N = 10), treated with vehicle [1:1:18, cremophor EL (polyoxyl 35 castor oil)/ethanol/saline]; Ctrl (N = 10), treated with saline. Animals were injected ip twice a day with 5 mg/kg body weight for 10 days. Lipid peroxidation, protein carbonylation and DNA oxidation were used as biomarkers of oxidative stress. The endogenous antioxidant defenses analyzed were glutathione (GSH) levels as well as enzyme activities of superoxide dismutase, catalase, glutathione S-transferase, glutathione reductase, and glutathione peroxidase (GPx) in liver homogenates. The levels of mRNA of the cannabinoid receptors CB1 and CB2 were also monitored. Treatment with Delta(9)-THC did not produce significant changes in oxidative stress markers or in mRNA levels of CB1 and CB2 receptors in the liver of mice, but attenuated the increase in the selenium-dependent GPx activity (Delta(9)-THC: 8%; VCtrl: 23% increase) and the GSH/oxidized GSH ratio (Delta(9)-THC: 61%; VCtrl: 96% increase), caused by treatment with the vehicle. Delta(9)-THC administration did not show any harmful effects on lipid peroxidation, protein carboxylation or DNA oxidation in the healthy liver of mice but attenuated unexpected effects produced by the vehicle containing ethanol/cremophor EL.