Effects of Bacillus subtilis and its metabolites on corn silage quality.

Cellulolytic micro-organisms are potent silage inoculants that decrease the fibrous content in silage and increase the fibre digestibility and nutritional value of silage. This study aimed to evaluate the effects of Bacillus subtilis CCMA 0087 and its enzyme ß-glucosidase on the nutritional value and aerobic stability of corn silage after 30 and 60 days of storage. We compared the results among silage without inoculant (SC) and silages inoculated with B. subtilis 8 log10 CFU per kg forage (SB8), 9 log10 CFU per kg forage (SB9) and 9·84 log10 CFU per kg forage + ß-glucosidase enzyme (SBE). No differences were observed in the levels of dry matter, crude protein and neutral detergent fibre due to the different treatments or storage times of the silos. Notably, the population of spore-forming bacteria increased in the SB9-treated silage. At 60 days of ensiling, the largest populations of lactic acid bacteria were found in silages treated with SB8 and SBE. Yeast populations were low for all silages, irrespective of the different treatments, and the presence of filamentous fungi was observed only in the SBE-treated silage. Among all silage treatments, SB9 treatment resulted in the highest aerobic stability.

The Effects of Testosterone Therapy Combined with Swimming Exercise on Adipose Tissue and Biochemical Parameters in Male Obese Wistar Rats.

Context: Exercise and anabolic steroids are anticipated to promote fat mass reduction and so to decrease the number of comorbidities related to excessive weight. Objective: The aim of this study was to verify the influence of aerobic exercise and the use of steroids on the accumulation of adipose tissue and on the biochemical limitations of Wistar rats nourished by a hypercaloric diet. Methods: Forty, young male Wistar rats were split into four groups: obese control (n=10), obese under treatment (n=10), obese under aerobic exercise (n=10) and obese under aerobic exercise and treatment (n=10). All animals were fed with a hypercaloric diet and animals under treatment received intramuscular testosterone. Body (weight and visceral fat) and blood (lipidogram, glucose, and liver enzymes) parameters were assessed. Results: The group treated with aerobic exercise and testosterone revealed a reduction in body weight and visceral, perirenal, retroperitoneal and epididymal fats, accompanied by the blood levels of glucose, lactate, LDL-cholesterol, HDL-cholesterol, and lactate dehydrogenase; following high-intensity physical activity. Conclusion: The results support the theory that the combination of steroids and physical activity reduces the side-effects of androgenic-anabolic hormones and conveys benefits to some constraints.

In Vitro and In Vivo Studies of the Trypanocidal Effect of Novel Quinolines.

Therapies for human African trypanosomiasis and Chagas disease, caused by Trypanosoma brucei and Trypanosoma cruzi, respectively, are limited, providing minimal therapeutic options for the millions of individuals living in very poor communities. Here the effects of 10 novel quinolines are evaluated in silico and by phenotypic studies using in vitro and in vivo models. Absorption, distribution, metabolism, excretion, and toxicity (ADMET) properties revealed that most molecules did not infringe on Lipinski's rules, which is a prediction of good oral absorption. These quinolines showed high probabilities of Caco2 permeability and human intestinal absorption and low probabilities of mutagenicity and of hERG1 inhibition. In vitro screens against bloodstream forms of T. cruzi demonstrated that all quinolines were more active than the reference drug (benznidazole [Bz]), except for DB2171 and DB2192, with five (DB2187, DB2131, DB2186, DB2191, and DB2217) displaying 50% effective concentrations (EC50s) of <3 µM (4-fold lower than that of Bz). Nine quinolines were more effective than Bz (2.7 µM) against amastigotes, showing EC50s ranging from 0.6 to 0.1 µM. All quinolines were also highly active in vitro against African trypanosomes, showing EC50s of ≤0.25 µM. The most potent and highly selective candidates for each parasite species were tested in in vivo models. Results for DB2186 were promising in mice with T. cruzi and T. brucei infections, reaching a 70% reduction of the parasitemia load for T. cruzi, and it cured 2 out of 4 mice infected with T. brucei DB2217 was also active in vivo and cured all 4 mice (100% cure rate) with T. brucei infection.

Antitrypanosomal Activity of Sterol 14α-Demethylase (CYP51) Inhibitors VNI and VFV in the Swiss Mouse Models of Chagas Disease Induced by the Trypanosoma cruzi Y Strain.

Chagas disease is a life-threatening infection caused by a variety of genetically diverse strains of the protozoan parasite Trypanosoma cruzi The current treatment (benznidazole and nifurtimox) is unsatisfactory, and potential alternatives include inhibitors of sterol 14α-demethylase (CYP51), the cytochrome P450 enzyme essential for the biosynthesis of sterols in eukaryotes and the major target of clinical and agricultural antifungals. Here we performed a comparative investigation of two protozoon-specific CYP51 inhibitors, VNI and its CYP51 structure-based derivative VFV, in the murine models of infection caused by the Y strain of T. cruzi The effects of different treatment regimens and drug delivery vehicles were evaluated in animals of both genders, with benznidazole serving as the reference drug. Regardless of the treatment scheme or delivery vehicle, VFV was more potent in both genders, causing a >99.7% peak parasitemia reduction, while the VNI values varied from 91 to 100%. Treatments with VNI and VFV resulted in 100% animal survival and 0% natural relapse after the end of therapy, though, except for the 120-day treatment schemes with VFV, relapses after three cycles of immunosuppression were observed in each animal group, and quantitative PCR analysis revealed a very light parasite load in the blood samples (sometimes below or near the detection limit, which was 1.5 parasite equivalents/ml). Our studies support further investigations of this class of compounds, including their testing against other T. cruzi strains and in combination with other drugs.

In Vitro and In Vivo Trypanosomicidal Action of Novel Arylimidamides against Trypanosoma cruzi.

Arylimidamides (AIAs) have been shown to have considerable biological activity against intracellular pathogens, includingTrypanosoma cruzi, which causes Chagas disease. In the present study, the activities of 12 novel bis-AIAs and 2 mono-AIAs against different strains ofT. cruziin vitroandin vivowere analyzed. The most active wasm-terphenyl bis-AIA (35DAP073), which had a 50% effective concentration (EC50) of 0.5 µM for trypomastigotes (Y strain), which made it 26-fold more effective than benznidazole (Bz; 13 µM). It was also active against the Colombiana strain (EC50= 3.8 µM). Analysis of the activity against intracellular forms of the Tulahuen strain showed that this bis-AIA (EC50= 0.04 µM) was about 100-fold more active than Bz (2 µM). The trypanocidal effect was dissociated from the ability to trigger intracellular lipid bodies within host cells, detected by oil red labeling. Both an active compound (35DAP073) and an inactive compound (26SMB060) displayed similar activation profiles. Due to their high selectivity indexes, two AIAs (35DAP073 and 35DAP081) were moved toin vivostudies, but because of the results of acute toxicity assays, 35DAP081 was excluded from the subsequent tests. The findings obtained with 35DAP073 treatment of infections caused by the Y strain revealed that 2 days of therapy induced a dose-dependent action, leading to 96 to 46% reductions in the level of parasitemia. However, the administration of 10 daily doses in animals infected with the Colombiana strain resulted in toxicity, preventing longer periods of treatment. The activity of the combination of 0.5 mg/kg of body weight/day 35DAP073 with 100 mg/kg/day Bz for 10 consecutive days was then assayed. Treatment with the combination resulted in the suppression of parasitemia, the elimination of neurological toxic effects, and survival of 100% of the animals. Quantitative PCR showed a considerable reduction in the parasite load (60%) compared to that achieved with Bz or the amidine alone. Our results support further investigations of this class with the aim of developing novel alternatives for the treatment of Chagas disease.

Mapping and validation of Xanthomonas citri subsp citri genes regulated by putative plant-inducible promoter box (PIP-box).

Citrus canker, caused by the Gram-negative bacterium Xanthomonas citri subsp citri (Xac), is a major disease affecting citriculture worldwide, because of the susceptibility of the host and the lack of efficient control methods. Previous studies have reported that some genes of phytopathogenic bacteria possess a consensus nucleotide sequence (TTCGC...N15...TTCGC) designated the "plant-inducible-promoter box" (PIP box) located in the promoter region, which is responsible for activating the expression of pathogenicity and virulence factors when the pathogen is in contact with the host plant. In this study, we mapped and investigated the expression of 104 Xac genes associated with the PIP box sequences using a macroarray analysis. Xac gene expression was observed during in vitro (Xac grown for 12 or 20 h in XAM1 induction medium) or in vivo (bacteria grown in orange leaves for 3 to 5 days) infection conditions. Xac grown in non-induction NB liquid medium was used as the control. cDNA was isolated from bacteria grown under the different conditions and hybridized to the macroarray, and 32 genes differentially expressed during the infection period (in vitro or in vivo induction) were identified. The macroarray results were validated for some of the genes through semi-quantitative RT-PCR, and the functionality of the PIP box-containing promoter was demonstrated by activating b-glucuronidase reporter gene activity by the PIP box-containing promoter region during Xac-citrus host interaction.

Different Therapeutic Outcomes of Benznidazole and VNI Treatments in Different Genders in Mouse Experimental Models of Trypanosoma cruzi Infection.

The lack of translation between preclinical assays and clinical trials for novel therapies for Chagas disease (CD) indicates a need for more feasible and standardized protocols and experimental models. Here, we investigated the effects of treatment with benznidazole (Bz) and with the potent experimental T. cruzi CYP51 inhibitor VNI in mouse models of Chagas disease by using different animal genders and parasite strains and employing distinct types of therapeutic schemes. Our findings confirm that female mice are less vulnerable to the infection than males, show that male models are less susceptible to treatment with both Bz and VNI, and thus suggest that male models are much more suitable for selection of the most promising antichagasic agents. Additionally, we have found that preventive protocols (compound given at 1 dpi) result in higher treatment success rates, which also should be avoided during advanced steps of in vivo trials of novel anti-T. cruzi drug candidates. Another consideration is the relevance of immunosuppression methods in order to verify the therapeutic profile of novel compounds, besides the usefulness of molecular diagnostic tools (quantitative PCR) to ascertain compound efficacy in experimental animals. Our study aims to contribute to the development of more reliable methods and decision gates for in vivo assays of novel antiparasitic compounds in order to move them from preclinical to clinical trials for CD.

A new alternative use for coffee pulp from semi-dry process to ß-glucosidase production by Bacillus subtilis.

UNLABELLED: Coffee is among the most preferred nonalcoholic drinks, and its consumption is distributed globally. During the coffee fruiting process, however, a large amount of waste is generated in the form of pulp, mucilage, husks, and water waste. The pulp and mucilage have the chemical composition to support the growth of micro-organisms and the production of value-added product. The aim was testify pulp coffee can be considered as carbon and inductor source for ß-glucosidase by Bacillus subtilis CCMA 0087. The response surface methodology (RSM) based on a central composite rotatable design (CCRD) was employed for this optimization. The methodology used in the optimization process was validated by testing the best conditions obtained and comparing them with the values predicted by the model. The highest ß-glucosidase production (22·59 UI ml(-1) ) was reached in 24 h of culturing at coffee pulp concentration of 36·8 g l(-1) , temperature of 36·6°C, and pH of 3·64. SIGNIFICANCE AND IMPACT OF THE STUDY: Countries whose economy is based on agricultural activities generate a great deal of liquid and solid waste. Thus, it is important to develop new alternatives for using this waste rather than disposing it in the environment. The production of enzymes, and particularly cellulase, is one such alternative. In this study, we proposed to produce ß-glucosidase production from pulp coffee extract using a Bacillus subtilis strain.

In vitro and in vivo studies of the biological activity of novel arylimidamides against Trypanosoma cruzi.

Fifteen novel arylimidamides (AIAs) (6 bis-amidino and 9 mono-amidino analogues) were assayed against Trypanosoma cruzi in vitro and in vivo. All the bis-AIAs were more effective than the mono-AIAs, and two analogues, DB1967 and DB1989, were further evaluated in vivo. Although both of them reduced parasitemia, protection against mortality was not achieved. Our results show that the number of amidino-terminal units affects the efficacy of arylimidamides against T. cruzi.

Differential expression of IGF family members in heat-stressed embryos produced in vitro from OPU-derived oocytes of Nelore (Bos indicus) and Holstein (Bos taurus) cows.

The IGF system is related to embryo quality. We aim to determine the effect of the heat stress on the mRNA expression of IGF1 and IGF2, IGFR1 and IGFR2, IGFBP2 and IGFBP4, and PAPPA in in vitro production (IVP) blastocysts from Nelore and Holstein after ovum pick up (OPU) to better understand the differences between these breeds. Oocytes from four Nelore and seven Holstein were collected in six OPU sessions. Following in vitro maturation and fertilization using six Nelore or Holstein sires, embryos were divided into control (cultured at 39°C) and heat stress (HS; exposed to 41°C for 9 h). Blastocysts were submitted to RNA extraction. The IGF1 expression was higher in blastocysts under HS in both breeds, and the expression of IGFBP2 and IGFBP4 was higher in Holstein blastocysts under HS. The high PAPPA expression and the low expression of IGFBP2 and IGFBP4 are associated with a more efficient degradation of IGFBPs, which results in greater IGF bioavailability in Nelore blastocysts and may contribute to the superior HS tolerance in Nelore, when compared to Holstein.

Isolation of the pituitary gonadotrophic α-subunit hormone of the giant amazonian fish: pirarucu (Arapaima gigas).

The cDNAs of the α-subunit of the pituitary gonadotrophic hormones (GTHα) of fish of the order Osteoglossiformes or the superorder Osteoglossomorpha have never been sequenced. For a better understanding the phylogenetic diversity and evolution of PGHα in fish and for future biotechnological synthesis of the gonadotrophic hormones (ag-FSH and ag-LH), of Arapaima gigas, one of the largest freshwater fishes of the world, its GTHα cDNA was synthesized by reverse transcriptase and the polymerase chain reaction starting from total pituitary RNA. The ag-GTHα-subunit was found to be encoded by 348 bp, corresponding to a protein of 115 amino acids, with a putative signal peptide of 24 amino acids and a mature peptide of 91 amino acids. Ten cysteine residues, responsible for forming 5 disulfide linkages, 2 putative N-linked glycosylation sites and 3 proline residues, were found to be conserved on the basis of the known sequences of vertebrate gonadotrophic hormones. Phylogenetic analysis, based on the amino acid sequences of 38 GTHα-subunits, revealed the highest identity of A. gigas with members of the Acipenseriformes, Anguilliformes, Siluriformes and Cypriniformes (87.1-89.5 %) and the lowest with Gadiformes and Cyprinodontiformes (55.0 %). The obtained phylogenetic tree agrees with previous analysis of teleostei, since A. gigas, of the order of Osteoglossiformes, appears as the sister group of Clupeocephala, while Elopomorpha forms the most basal group of all other teleosts.

The trypanocidal activity of amidine compounds does not correlate with their binding affinity to Trypanosoma cruzi kinetoplast DNA.

Due to limited efficacy and considerable toxicity, the therapy for Chagas' disease is far from being ideal, and thus new compounds are desirable. Diamidines and related compounds such as arylimidamides have promising trypanocidal activity against Trypanosoma cruzi. To better understand the mechanism of action of these heterocyclic cations, we investigated the kinetoplast DNA (kDNA) binding properties and trypanocidal efficacy against T. cruzi of 13 compounds. Four diamidines (DB75, DB569, DB1345, and DB829), eight arylimidamides (DB766, DB749, DB889, DB709, DB613, DB1831, DB1852, and DB2002), and one guanylhydrazone (DB1080) were assayed in thermal denaturation (T(m)) and circular dichroism (CD) studies using whole purified T. cruzi kDNA and a conserved synthetic parasite sequence. The overall CD spectra using the whole kDNA were similar to those found for the conserved sequence and were indicative of minor groove binding. Our findings showed that some of the compounds that exhibited the highest trypanocidal activities (e.g., DB766) caused low or no change in the T(m) measurements. However, while some active compounds, such as DB766, induced profound alterations of kDNA topology, others, like DB1831, although effective, did not result in altered T(m) and CD measurements. Our data suggest that the strong affinity of amidines with kDNA per se is not sufficient to generate and trigger their trypanocidal activity. Cell uptake differences and possibly distinct cellular targets need to be considered in the final evaluation of the mechanisms of action of these compounds.

Biological control in the germination of seeds from two species native of the Cerrado region.

Microorganisms have been efficiently used for the biological control of phytopathogens through the production of antimicrobial substances. However, the objectives of this work were: to study the germination of Butia purpurascens Glassman and Butia archeri Glassman seeds in different substrates, to select and identify the endophytic and rhizospheric bacterial isolates of B. purpurascens and B. archeri, and to perform antibiosis tests based on the isolated microorganisms of these tree species. No difference was found between the cultivation substrates for the percentages of germination, hard seeds, and fungal contamination in the B. purpurascens seeds. The Bacillus subtilis isolated showed the best capacity for suppressing the growth of the two deteriorative fungi tested in B. purpurascens seeds. No difference was found for inhibition of the growth of Aspergillus niger fungus (deteriorative fungus of B. archeri seeds) between the microorganisms with Bacillus sp. and Brevibacillus brevis compared to the control. In the microbiolization of B. purpurascens and B. archeri seeds performed with microbiological solutions produced from the endophytic and rhizospheric strains of Bacillus sp., no differences were observed in the percentages of germination and contamination by fungi. For B. archeri seeds, there was contamination by fungi and bacteria after one day of cultivation, primarily in the regions with lesions caused by the extraction and scarification process.

Features of Saccharomyces cerevisiae as a culture starter for the production of the distilled sugar cane beverage, cachaça in Brazil.

AIMS: To evaluate the dominance and persistence of strains of Saccharomyces cerevisiae during the process of sugar cane fermentation for the production of cachaça and to analyse the microbial compounds produced in each fermentative process. METHODS AND RESULTS: Three S. cerevisiae strains were evaluated during seven consecutive 24-h fermentation batches using recycled inocula. The UFLA CA 116 strain had the largest population of viable organisms, and the maximum population was achieved in the fourth batch after 96 h of fermentation. The UFLA CA 1162 and UFLA CA 1183 strains grew more slowly, and the maximum population was reached in the seventh batch. Molecular characterization of isolated yeast cells using PFGE (pulse field gel electrophoresis) revealed that more than 86% of the isolates corresponded to the initially inoculated yeast strain. The concentration of aldehydes, esters, methanol, alcohol and volatile acids in the final-aged beverages were within the legal limits. CONCLUSIONS: Cachaça produced by select yeast strains exhibits analytical differences. UFLA CA 1162 and UFLA CA 116 S. cerevisiae isolates can be considered the ideal strains for the artisanal production of cachaça in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of select yeast strains can improve the quality and productivity of cachaça production. Our findings are important for the appropriate monitoring of yeast during sugar cane fermentation. In addition, we demonstrate that UFLA CA 116 and UFLA CA 1162, the ideal yeast strains for cachaça production, are maintained at a high population density. The persistence of these yeast strains in the fermentation of sugar cane juice promotes environmental conditions that prevent or decrease bacterial contamination. Thus, the use of select yeast strains for the production of cachaça is a viable economic alternative to standardize the production of this beverage.

Double membrane based on lidocaine-coated polymyxin-alginate nanoparticles for wound healing: In vitro characterization and in vivo tissue repair.

The aim of this study was to develop and characterize a double layer biomembrane for dual drug delivery to be used for the treatment of wounds. The membrane was composed of chitosan, hydroxypropyl methylcellulose and lidocaine chloride (anesthetic drug) in the first layer, and of sodium alginate-polymyxin B sulphate (antibiotic) nanoparticles as the second layer. A product with excellent thickness (0.01-0.02 mm), adequate mechanical properties with respect to elasticity, stiffness, tension, and compatible pH for lesion application has been successfully obtained. The incorporation of the drugs was confirmed analysing the membrane cross-sections by scanning electron microscopy. A strong interaction between the drugs and the functional groups of respective polymers was confirmed by Fourier-Transform Infrared Spectroscopy, thermal analysis and X-ray diffraction. Microbiological assays showed a high antimicrobial activity when polymyxin B was present to act against the Staphylococcus aureus and Pseudomonas aeruginosa strains. Low cytotoxicity observed in a cell viability colorimetric assay and SEM analysis suggest biocompatibility between the developed biomembrane and the cell culture. The in vivo assay allowed visualizing the healing potential by calculating the wound retraction index and by histological analysis. Our results confirm the effectiveness of the developed innovative biomaterial for tissue repair and regeneration in an animal model.

Silver Nanoparticles-Composing Alginate/Gelatine Hydrogel Improves Wound Healing In Vivo.

Polymer hydrogels have been suggested as dressing materials for the treatment of cutaneous wounds and tissue revitalization. In this work, we report the development of a hydrogel composed of natural polymers (sodium alginate and gelatin) and silver nanoparticles (AgNPs) with recognized antimicrobial activity for healing cutaneous lesions. For the development of the hydrogel, different ratios of sodium alginate and gelatin have been tested, while different concentrations of AgNO3 precursor (1.0, 2.0, and 4.0 mM) were assayed for the production of AgNPs. The obtained AgNPs exhibited a characteristic peak between 430-450 nm in the ultraviolet-visible (UV-Vis) spectrum suggesting a spheroidal form, which was confirmed by Transmission Electron Microscopy (TEM). Fourier Transform Infra-red (FT-IR) analysis suggested the formation of strong intermolecular interactions as hydrogen bonds and electrostatic attractions between polymers, showing bands at 2920, 2852, 1500, and 1640 cm-1. Significant bactericidal activity was observed for the hydrogel, with a Minimum Inhibitory Concentration (MIC) of 0.50 µg/mL against Pseudomonas aeruginosa and 53.0 µg/mL against Staphylococcus aureus. AgNPs were shown to be non-cytotoxic against fibroblast cells. The in vivo studies in female Wister rats confirmed the capacity of the AgNP-loaded hydrogels to reduce the wound size compared to uncoated injuries promoting histological changes in the healing tissue over the time course of wound healing, as in earlier development and maturation of granulation tissue. The developed hydrogel with AgNPs has healing potential for clinical applications.

Biodegradable antioxidant chitosan films useful as an anti-aging skin mask.

Cosmetics, personal care and biomedical products obtained by bio-based polymers and natural bioactive compounds are a new growing market. The ecological awareness is changing consumers' demands, causing consumers to look for more sustainable options, with a reduced environmental impact. The innovation of this work was to develop a natural polymer matrix (chitosan) entrapping antioxidant actives compounds such as annatto (Bixa Orellana L.) and vitamin C with potential application as sustainable anti-aging skin mask treatment. Films of chitosan (Ch) and reacetylated chitosan (RCh), exhibiting different degrees of acetylation (DA = 13.3 and 33.9%, respectively), were produced. The formulations of active films of chitosan (BCh) and reacetylated chitosan (BRCh) were 1% (w/w) of chitosan, 1% (w/w) of annatto powder, 5% (w/w) of vitamin C and 1% (w/w) of glycerol (as plasticizer). Reacetylated chitosan films (DA = 33.9%) presented higher water affinity than chitosan films (DA = 13.3%). The elongation of RCh and BRCh increased and the resistance decreased, as compared to Ch and BCh. The antioxidants compounds (annatto and vitamin C) of BRCh films released faster than BCh films. Thus, the BRCh films showed potential application as an anti-aging skin mask.

Microdialysis as a tool to determine free kidney levels of voriconazole in rodents: a model to study the technique feasibility for a moderately lipophilic drug.

Microdialysis has been employed for the in vivo measurement of endogenous compounds and a variety of drugs in different tissues. The applicability of this technique can be limited by drug lipophilicity which can impair the diffusion through dialysis membrane. The objective of this study was to evaluate the feasibility of using microdialysis to study kidney penetration of voriconazole, a moderately lipophilic antifungal triazolic agent (LogD7.4=1.8). Microdialysis probe recoveries were investigated in vitro by dialysis and retrodialysis using four different drug concentrations (0.1-2 microg/mL) at five flow rates (1-5 microL/min). Recoveries were dependent on the method used for the determination as well as on the flow rate, but independent of drug concentration. The average apparent recoveries determined by dialysis and retrodialysis, at flow rate of 2 microL/min, were 21.1+/-1.5% and 28.7+/-2.0%, respectively. Recovery by retrodialysis was bigger than the recovery by dialysis. The average apparent dialysis/retrodialysis recovery ratio in vitro was 0.73 for all concentrations investigated. The differences between retrodialysis and dialysis recoveries were attributed to the drug's binding to the plastic tubing before and after the dialysis membrane which was experimentally evaluated and mathematically modeled. The in vivo apparent recovery determined by retrodialysis in healthy Wistar rats' kidney was 38.5+/-3.5%, similar to that observed in vitro using the same method (28.7+/-2.0%). The in vivo apparent recovery after correcting for plastic tubing binding (25.1+/-2.8%) was successfully used for determining free kidney levels of voriconazole in rats following 40 and 60mg/kg oral dosing. The results confirmed that microdialysis can be used as sampling technique to determine free tissue levels of moderately lipophilic drugs once the contribution of tubing binding and membrane diffusion on the apparent recovery are disentangled.

Event-related potentials modulated by the perception of sexual dimorphism: The influence of attractiveness and sex of faces.

Sexual dimorphism has been proposed as one of the facial traits to have evolved through sexual selection and to affect attractiveness perception. Even with numerous studies documenting its effect on attractiveness and mate choice, the neurophysiological correlates of the perception of sexual dimorphism are not yet fully understood. In the present study, event-related potentials (ERPs) were recorded during visualisation of faces that had been previously transformed in shape to appear more masculine or more feminine. The participants' task consisted of judging the attractiveness of half of the total number of faces, and performing a sex discrimination task on the other half. Both early and late potentials were modulated by the sex of faces, whereas the effect of the sexually dimorphic transform was mainly visible in the P2 (positive deflection around 200 ms after stimulus onset), EPN (early posterior negativity) and LPP (late positive potential) components. There was an effect of sexual dimorphism on P2 and EPN amplitudes when female participants visualised male faces, which may indicate that masculinity is particularly attended to when viewing opposite sex members. Also, ERP results seem to support the idea of sex differences in social categorisation decisions regarding faces, although differences were not evident on behavioural results. In general, these findings contribute to a better understanding of how humans perceive sexually dimorphic characteristics in other individuals' faces and how they affect attractiveness judgements.