In silico prediction of the enzymes involved in the degradation of the herbicide molinate by Gulosibacter molinativorax ON4T.

Gulosibacter molinativorax ON4T is the only known organism to produce molinate hydrolase (MolA), which catalyses the breakdown of the thiocarbamate herbicide into azepane-1-carboxylic acid (ACA) and ethanethiol. A combined genomic and transcriptomic strategy was used to fully characterize the strain ON4T genome, particularly the molA genetic environment, to identify the potential genes encoding ACA degradation enzymes. Genomic data revealed that molA is the only catabolic gene of a novel composite transposon (Tn6311), located in a novel low copy number plasmid (pARLON1) harbouring a putative T4SS of the class FATA. pARLON1 had an ANI value of 88.2% with contig 18 from Agrococcus casei LMG 22410T draft genome. Such results suggest that pARLON1 is related to genomic elements of other Actinobacteria, although Tn6311 was observed only in strain ON4T. Furthermore, genomic and transcriptomic data demonstrated that the genes involved in ACA degradation are chromosomal. Based on their overexpression when growing in the presence of molinate, the enzymes potentially involved in the heterocyclic ring breakdown were predicted. Among these, the activity of a protein related to caprolactone hydrolase was demonstrated using heterologous expression. However, further studies are needed to confirm the role of the other putative enzymes.

Discovery of serum biomarkers for diagnosis of tuberculosis by NMR metabolomics including cross-validation with a second cohort.

BACKGROUND: Tuberculosis (TB) is a disease with worldwide presence and a major cause of death in several developing countries. Current diagnostic methodologies often lack specificity and sensitivity, whereas a long time is needed to obtain a conclusive result. METHODS: In an effort to develop better diagnostic methods, this study aimed at the discovery of a biomarker signature for TB diagnosis using a Nuclear Magnetic Resonance based metabolomics approach. In this study, we acquired 1H NMR spectra of blood serum samples of groups of healthy subjects, individuals with latent TB and of patients with pulmonary and extra-pulmonary TB. The resulting data were treated with uni- and multivariate statistical analysis. RESULTS: Six metabolites (inosine, hypoxanthine, mannose, asparagine, aspartate and glutamate) were validated by an independent cohort, all of them related with metabolic processes described as associated with TB infection. CONCLUSION: The findings of the study are according with the WHO Target Product Profile recommendations for a triage test to rule-out active TB.

Impairment of swimming performance in Tritia reticulata (L.) veligers under projected ocean acidification and warming scenarios.

Tritia reticulata (L.) is a neogastropod ubiquitous in the coastal communities of the NE Atlantic. Its life cycle relies on the swimming performance of planktonic early life stages, whose sensitivity to the climate conditions projected for the near future, namely of ocean acidification (OA) and warming (W), is, to our best knowledge, unknown. To examine the resilience of larval stages to future environmental conditions, this work investigates the effect of OA-W on the swimming performance of T. reticulata veligers under a range of experimental conditions, based on the end-of-century projections of the Intergovernmental Panel on Climate Change. Veligers were exposed to six experimental scenarios for 14 days, employing a full factorial design with three temperatures (T°C: 18, 20 and 22 °C) and two pH levels (pHtarget: 8.1 and 7.8). Mortality was assessed throughout the trial, after which swimming behaviour - characterised by the activity, speed and the distance travelled by veligers - was analysed by automated video recordings in a Zebrabox® device. Mortality increased with OA-W and, although more active, larvae travelled shorter distances revealing reduced swimming speed under acidic and warmer conditions, with the interaction of the tested stressors - pH and T°C - being highly significant. Results motivated the morpho-histological analysis of larvae preserved at the end of the trial, to check for the integrity of the organs involved in veligers' motion: statocysts, velum and foot. Statocyst and velar morpho-structure were conserved but histological damage of metapodial epithelia was evident under acidity, namely an apparent hypertrophy and protrusion of the secretory cells, with dispersed pigmented granules and, at 22 °C, less cilia, with potential functional implications. Negative consequences of the OA-W scenarios tested on veligers' competence are unveiled, pointing towards the eminent threat these phenomena constitute to T. reticulata perpetuation in case no mitigation measures are taken, and projections become effective.

Discriminant ability of phenolic compounds and short chain organic acids profiles in the determination of quality parameters of Brandy de Jerez.

Brandy de Jerez is the most produced spirit in Spain. The rules of its Regulatory Council require the spirit to age in American oak casks that have previously contained any kind of sherry wine. This use, called seasoning, releases wine compounds into the spirit. Because of the differences among sherries, the organoleptic features of a brandy will be significantly different from any other depending on the seasoning. In addition, its specific features make it different from any other spirit. The chromatographic profiles of Brandy de Jerez are reported to be different depending on the seasonings through their ageing process. Different types of Brandy de Jerez have been characterised, regarding their seasoning, using chromatographic techniques. Applying statistical analysis, correlations between the chromatographic profiles and the seasonings have risen up. In addition, the profiles have demonstrated to possess a high degree of correlation with the ageing time of the samples.

Fast Determination of Phenolic Compounds in Rice Grains by Ultraperformance Liquid Chromatography Coupled to Photodiode Array Detection: Method Development and Validation.

There are several phenolic compounds in rice grains providing benefits for human health. The concentration of phenolic compounds in rice is strongly affected by the polishing steps during rice production. A new sensitive ultraperformance liquid chromatography-ultraviolet-visible spectroscopy method with a photodiode array detection protocol has been developed and validated for the quantitation of phenolic compounds in rice grains. Several working variables and two different columns were evaluated. Finally, a less than 3 min analysis time was developed to achieve enough resolution for the simultaneous determination of the 20 most common phenolic compounds in rice. The analytical properties for the separation method produced an adequate sensitivity for all phenolic compounds in the regular range for phenolics in rice, 0.5-100 mg L-1 ( R2 > 0.997), with high precisions for both repeatability and intermediate precisions (coefficients of variation less than 0.4 and 2.5% for the retention time and area of the peaks, respectively).

Thymol action on cells and tissues of the synganglia and salivary glands of Rhipicephalus sanguineus sensu lato females (Acari: Ixodidae).

Thymol is a monoterpene present in plants of the families Lamiaceae, Verbenaceae and Apiaceae. Despite its proven acaricidal activity, little is known about the mechanism of action of thymol in ticks. Thus, the aim of this study was to perform a morpho-histochemical analysis of the synganglion and salivary glands of partially engorged females of the brown dog tick, Rhipicephalus sanguineus sensu lato (s.l.), exposed to thymol at different concentrations. Five groups were established: Control Group I (distilled water), Control Group II (ethanol 30%), Group III (thymol 1.25 mg/mL), Group IV (thymol 2.5 mg/mL) and Group V (thymol 5.0 mg/mL). The females were exposed to the treatments by the immersion method and subsequently kept in a climatic chamber (27 ± 1 °C and relative humidity 80 ± 10%) for five days. After this period, the synganglion and salivary glands were removed, and the hematoxylin/eosin morphological technique was applied. The von Kossa staining method with counterstaining neutral red was performed on the salivary glands. The results showed that females exposed to thymol had damaged synganglia, with pyknotic nuclei and vacuoles in the cortex and subperineurial regions, as well as rupture of the neural lamellae. The salivary glands showed type I acini with a dilated lumen. Cells with extremely vacuolated cytoplasm and fragmented nuclei were observed in type II and III acini. Type II acini of the females exposed to thymol revealed different calcium staining when compared to the Control Groups I and II. We therefore conclude that the salivary glands and synganglion are subject to changes in morphology and calcium levels when exposed to thymol at concentrations of 1.25, 2.5 and 5.0 mg/mL, demonstrating that this monoterpene has acaricidal potential on partially engorged females of R. sanguineus (s.l.).

Study of a laboratory-scaled new method for the accelerated continuous ageing of wine spirits by applying ultrasound energy.

During the ageing of brandies, many physicochemical processes take place involving the distilled spirit and the wood of the casks. Because of these reactions, the polyphenolic content of brandies and their content of organic acids increase with the ageing. These reactions are slow, and the ageing of high-quality brandies takes several years. In this paper, the development of a system that uses the circulation of the wine distillate through encapsulated American oak chips and the application of ultrasound energy with the aim of producing aged wine spirits has been carried out, and the influences of the operation variables over the characteristics of the produced drink have been measured. With that proposal, the influence of different powers of ultrasound, and also the influence of the movement of the liquor through oak chips, was determined first. This way, the results show that higher powers of ultrasound, of nearly 40W/L, in addition with the movement of the spirit, improve the extraction of phenolic compounds in a 33.94%, after seven days of ageing. Then, applying Youden and Steiner's experimental design, eight experiments of ageing were performed, and the samples obtained by this new method were analysed to obtain information related to their physicochemical and oenological characterisation in order to determine the experimental conditions that produce the best ageing results. This way, the best spirit produced by this new method of ageing is obtained with a high alcoholic strength of the distilled wine and a high quantity of oak chips, and with room temperature and high flow rate. In addition, the presence of oxygen in the sample and the absence of light increase the quality of the produced spirit. Finally, the application of ultrasound energy in large pulses is related with the improvement of two important ageing markers: the intensity of the colour and the TPI. As a last experiment, we applied this ageing method to five varietal spirits. The sensorial analysis of aged samples showed the aged spirits had better ratings than the initial distilled wine.

Pressurized liquid extraction of phenolic compounds from rice (Oryza sativa) grains.

An analytical pressurized liquid extraction (PLE) process has been studied for the extraction of phenolic compounds from rice grains. A fractional factorial design (2(7-2)) with a centre point was used to optimize PLE parameters such as solvent composition (EtOAc in MeOH), extraction temperature, pressure, flushing, static extraction time, solvent-purge and sample weight. Extraction temperature, solvent and static extraction time were found to have a significant effect on the response value. The optimized method was validated for selectivity, linearity, limits of detection and quantification, recovery and precision. The validated method was successfully applied for the analysis of a wide variety of rice grains. Seventeen phenolic compounds were detected in the sample and guaiacol, ellagic acid, vanillic acid and protocatechuic acid were identified as the most abundant compounds. Nonetheless, different species of rice show very varied compound diversity and levels of compounds in their grain compositions.

Concomitant histone deacetylase and phosphodiesterase 5 inhibition synergistically prevents the disruption in synaptic plasticity and it reverses cognitive impairment in a mouse model of Alzheimer's disease.

BACKGROUND: Given the implication of histone acetylation in memory processes, histone deacetylase inhibitors (HDACIs) have been postulated as potential modulators of cognitive impairment in Alzheimer's disease (AD). However, dose-dependent side effects have been described in patients with the currently available broad-spectrum HDACIs, explaining why their therapeutic potential has not been realized for chronic diseases. Here, by simultaneously targeting two independent enzyme activities, histone deacetylase (HDAC) and phosphodiesterase-5 (PDE5), we propose a novel mode of inhibitory action that might increase the therapeutic specificity of HDACIs. RESULTS: The combination of vorinostat, a pan-HDACI, and tadalafil, a PDE5 inhibitor, rescued the long-term potentiation impaired in slices from APP/PS1 mice. When administered in vivo, the combination of these drugs alleviated the cognitive deficits in AD mice, as well as the amyloid and tau pathology, and it reversed the reduced dendritic spine density on hippocampal neurons. Significantly, the combination of vorinostat and tadalafil was more effective than each drug alone, both against the symptoms and in terms of disease modification, and importantly, these effects persisted after a 4-week washout period. CONCLUSIONS: The results highlight the pharmacological potential of a combination of molecules that inhibit HDAC and PDE5 as a therapeutic approach for AD treatment.

Comparison of the international normalized ratio levels obtained by portable coagulometer and laboratory in a clinic specializing in oral anticoagulation.

INTRODUCTION: Therapy with Vitamin K antagonists (VKA) is effective in reducing thromboembolic events in various diseases. There are limitations however, which limit clinical handling and maintaining INR within therapeutic range. Studies have shown that portable coagulometers, when compared to laboratory tests, are more practical and provide better patient adherence and involvement toward treatment which results in better INR control. This study aimed to evaluate laboratory obtained INR results compared to two different portable coagulometers. METHODS: A prospective study which monitored 1009 patients using VKA in the Anticoagulation Clinic at the Institute Dante Pazzanese of Cardiology in São Paulo between July and September 2012. Patient INR values were obtained by the laboratory through venipuncture and then compared to INR values obtained by capillary puncture from two different portable coagulometers. RESULTS: Overall, 1009 patients were included in the study; among these, 520 (51.5%) are male with average age of 59.6 years (13-91). The more common indications were atrial fibrillation (49.9%) and mechanical prosthesis (33.7%). The correlation coefficient was of 0.95 with and 0.88 with INRatio PT Monitor(®) compared to laboratory. In patients with INR < 2 (lower than therapeutic range), the coefficient was 0.92 and 0.81 for CoaguChek XS plus(®) and INRatio PT Monitor(®) respectively. In patients within therapeutic range (INR 2-3), the coefficient was 0.86 with CoaguChek XS Plus(®) and 0.76 with INRatio PT Monitor(®) . For INR above therapeutic range (INR > 3.0) the correlation was 0.80 with CoaguChek XS Plus(®) and 0.54 with INRatio PT Monitor(®) . As for concordance between methods, the intraclass correlation coefficients (ICC) were slightly smaller than those previously stated (ICC = 0.899 with CoaguChek XS Plus(®) and ICC = 0.716 with INRatio PT Monitor(®) ). CONCLUSION: The use of portable coagulometers was comparable to laboratory tests and better correlation coefficients were observed with CoaguChek XS Plus(®) and in patients with INR lower or within therapeutic range. Portable coagulometers proved to be a useful and reliable tool for INR control in patients using VKA.

Optimisation and validation of the microwave-assisted extraction of phenolic compounds from rice grains.

A new microwave-assisted extraction (MAE) method has been investigated for the extraction of phenolic compounds from rice grains. The experimental conditions studied included temperature (125-175°C), microwave power (500-1000W), time (5-15min), solvent (10-90% EtOAc in MeOH) and solvent-to-sample ratio (10:1 to 20:1). The extraction variables were optimised by the response surface methodology. Extraction temperature and solvent were found to have a highly significant effect on the response value (p<0.0005) and the extraction time also had a significant effect (p<0.05). The optimised MAE conditions were as follows: extraction temperature 185°C, microwave power 1000W, extraction time 20min, solvent 100% MeOH, and solvent-to-sample ratio 10:1. The developed method had a high precision (in terms of CV: 5.3% for repeatability and 5.5% for intermediate precision). Finally, the new method was applied to real samples in order to investigate the presence of phenolic compounds in a wide variety of rice grains.

Comparison of the international normalized ratio levels obtainedby portable coagulometer and laboratory in a clinic specializing inoral anticoagulation

Therapy with Vitamin K antagonists (VKA) is effectivein reducing thromboembolic events in various diseases. There arelimitations however, which limit clinical handling and maintainingINR within therapeutic range. Studies have shown that portable coagulometers,when compared to laboratory tests, are more practicaland provide better patient adherence and involvement towardtreatment which results in better INR control. This study aimed toevaluate laboratory obtained INR results compared to two differentportable coagulometers.Methods: A prospective study which monitored 1009 patients usingVKA in the Anticoagulation Clinic at the Institute Dante Pazzaneseof Cardiology in S~ao Paulo between July and September 2012.Patient INR values were obtained by the laboratory through venipunctureand then compared to INR values obtained by capillarypuncture from two different portable coagulometers.Results: Overall, 1009 patients were included in the study; amongthese, 520 (51.5%) are male with average age of 59.6 years (13–91). The more common indications were atrial fibrillation (49.9%)and mechanical prosthesis (33.7%). The correlation coefficient wasof 0.95 with and 0.88 with INRatio PT Monitor compared to laboratory.In patients with INR 3.0) the correlation was 0.80 with CoaguChek XSPlus and 0.54 with INRatio PT Monitor ...

Relación entre los Niveles de Fibrinógeno e Insulina con los Parámetros que Definen al Síndrome Metabólico / Relationship between Insulin levels with Fibrinogen and parameters defining the Metabolic Syndrome

Metabolic syndrome is a combination of risk factors that increase an individual's chance of developing ischemic cardiovascular diseases which are the leading cause of death worldwide. In a number of metabolic syndrome abnormalities such as impaired secretion of acute phase reactants and certain hormones, such as the case of fibrinogen and insulin respectively. In this research fibrinogen levels were correlated with insulin and the factors that define the metabolic syndrome according to NCEP- ATP III in a population from the Municipality Francisco Linares Alcántara aged 35 and 60, the sample consisted of 30 patients and 29 controls. Was made comparison of means of anthropometric parameters and biochemical testing using the Wilcoxon signed ranks with a statistical significance (p < 0.05) and Spearman correlation analysis between insulin and fibrinogen levels with anthropometric and biochemical parameters studied was made it was found that patients had higher BMI, glucose , triglycerides, VLDL-C , insulin and fibrinogen compared to controls. Furthermore, the latter showed the highest HDL-C value compared with patients . On the other hand there was a positive association between insulin levels with BMI and glycemic variables . There was a negative association between levels of fibrinogen and LDL cholesterol variables in patients. No association between insulin levels and fibrinogen was found.

El síndrome metabólico es la conjunción de factores de riesgo en un individuo que aumentan su probabilidad de padecer enfermedades cardiovasculares isquémicas las cuales constituyen la primera causa de muerte a nivel mundial. En el síndrome metabólico se desencadenan una serie de anomalías como ejemplo alteración en la secreción de reactantes de fase aguda y de ciertas hormonas, como el caso del fibrinógeno y la insulina respectivamente. En esta investigación se correlacionaron los niveles de fibrinógeno y la insulina con los factores que definen al síndrome metabólico según la NCEP-ATP III en una población procedente del Municipio Francisco Linares Alcántara en edades comprendidas entre 35 y 60 años. La muestra estaba conformada entre 30 pacientes y 29 controles. Se hizo comparación de medias de los parámetros antropométricos y bioquímicos utilizando la prueba de los rangos con signo de Wilcoxon con una significancia estadística de (p<0,05). De igual modo se realizó análisis de correlación de Spearman entre los niveles de Insulina y fibrinógeno con los parámetros antropométricos y bioquímicos estudiados se encontró que los pacientes tenían más elevado el IMC, glicemia, triglicéridos, VLDLc, insulina y fibrinógeno con respecto a los controles. Además, estos últimos presentaron el valor de HDLc más elevado comparado con el de los pacientes. Por otro lado hubo asociación positiva entre los niveles de insulina con las variables IMC y glicemia. Hubo asociación negativa entre los niveles de fibrinógeno con las variables colesterol y LDLc en los pacientes. No se encontró asociación entre los niveles de insulina y fibrinógeno.

Complete genome sequence of the Radiation-Resistant bacterium Rubrobacter radiotolerans RSPS-4.

Rubrobacter radiotolerans strain RSPS-4 is a slightly thermophilic member of the phylum "Actinobacteria" isolated from a hot spring in São Pedro do Sul, Portugal. This aerobic and halotolerant bacterium is also extremely resistant to gamma and UV radiation, which are the main reasons for the interest in sequencing its genome. Here, we present the complete genome sequence of strain RSPS-4 as well as its assembly and annotation. We also compare the gene sequence of this organism with that of the type strain of the species R. radiotolerans isolated from a hot spring in Japan. The genome of strain RSPS-4 comprises one circular chromosome of 2,875,491 bp with a G+C content of 66.91%, and 3 circular plasmids of 190,889 bp, 149,806 bp and 51,047 bp, harboring 3,214 predicted protein coding genes, 46 tRNA genes and a single rRNA operon.

Effects of a diabetes prevention programme on weight-specific quality of life in Latino youth.

OBJECTIVE: To examine the effects of a diabetes prevention programme on weight-specific Quality of Life (QOL) in obese Latino youth. METHODS: Fifteen obese Latino adolescents (body mass index % = 96.4 ± 1.2; age = 15.0 ± 1.0) completed a 12-week culturally grounded, community-based intervention designed to improve physical and psychosocial health. Weight-specific QOL was assessed by the Youth Quality of Life-Weight module and compared with age, sex and ethnicity-matched lean youth. RESULTS: At baseline, intervention youth exhibited significantly lower weight-specific QOL compared with lean youth (70.8 ± 5.4 vs. 91.2 ± 2.2, P = 0.002). However, following the intervention, total weight-specific QOL increased by 21.8% among obese youth (70.8 ± 5.4 to 86.2 ± 4.3, P < 0.001) and was no longer different from lean controls. Significant increases in weight-specific QOL were noted across all subdomains including self (45.7%), social (11.9%) and environmental (36.2%) despite the fact that weight did not change (90.6 ± 6.8 to 89.9 ± 7.2, P = 0.44). The improvements in QOL were maintained for up to 12 months after the intervention. CONCLUSION: Weight-specific QOL among obese Latino youth can be improved through lifestyle interventions to a level similar to lean peers. Further, weight loss may not be necessary to observe improvements in QOL.

Assessment of imposex and butyltin concentrations in Gemophos viverratus (Kiener, 1834), from São Vicente, Republic of Cabo Verde (Africa).

This work constitutes the first assessment of tributyltin (TBT) pollution levels in the Republic of Cabo Verde (Africa) and proposes the marine gastropod Gemophos viverratus (Kiener, 1834) as a new bioindicator of TBT pollution in the Macaronesia and west coast of Africa. Specimens were collected between August and October 2012 along a gradient of naval traffic in São Vicente Island. The results clearly indicate an increase of imposex levels (percentage of females affected with imposex, 0-100 %; vas deferens sequence index, 0-4.1; relative penis length index, 0-54.6 %) and female TBT contamination (from 5 to 37 ngSn g(-1) dry weight (dw)) from outside to inside the harbour of Porto Grande Bay and identify this area as the focus of TBT pollution in the island. The butyltin degradation index for G. viverratus tissues ranged between 1.3 and 2.2, which being above 1 suggests that a considerable part of TBT inputs to the bay may not be very recent. Sterile females were found inside the harbour with an incidence up to 21.4 %. Considering the existence of a planktonic veliger stage in the life cycle of G. viverratus, it is expected that recruitment of newborn individuals can be supplied from unaffected breeding females inside and outside the Porto Grande Bay, resulting in a reduced impact of TBT pollution on population abundance. G. viverratus is very promising to be used as a simple, inexpensive and efficient novel tool for TBT pollution biomonitoring in the Macaronesia and west coast of Africa, a region for which there is an astonishing lack of information concerning levels and ecological impacts of TBT pollution.

Direct determination of organic acids in wine and wine-derived products by Fourier transform infrared (FT-IR) spectroscopy and chemometric techniques.

FT-IR with partial least squares (PLS) was used to establish a full calibration model for tartaric acid, malic acid, lactic acid, succinic acid, citric acid and acetic acid in wines, vinegars and spirits. Sample pre-treatment was not required except for filtering. The PLS method was employed and FT-IR spectra were correlated with the results from a reference HPLC method. In the validation with an independent set of samples, a strong correlation with the reference values was demonstrated for the highest concentration range (>0.6 g L(-1)) in all acids but the correlation was much weaker in the lower range (<0.6 g L(-1)). In the case of acetic acid, however, good results were obtained in the low concentration range for both red and white wine and for spirit drinks. This finding explains the fact that calibration and validation of the FT-IR spectroscopy method depends very strongly on the composition of the sample set and on the quality of the reference analysis. It was not possible to obtain a single calibration for all of the analysed samples and in some cases individual calibrations for specific samples were required. This situation was due to the different matrixes in the studied samples: 12-15% ethanol (wines), 30-40% ethanol (spirits) and 6-10% acetic acid (vinegars). As a result, a calibration model was developed for each acid in red and white wine, tartaric acid, acetic acid and total acidity in vinegar, and acetic acid in spirit drinks.

Differential genotoxicity of Roundup(®) formulation and its constituents in blood cells of fish (Anguilla anguilla): considerations on chemical interactions and DNA damaging mechanisms.

It has been widely recognized that pesticides represent a potential threat in aquatic ecosystems. However, the knowledge on the genotoxicity of pesticides to fish is still limited. Moreover, genotoxic studies have been almost exclusively focused on the active ingredients, whereas the effect of adjuvants is frequently ignored. Hence, the present study addressed the herbicide Roundup®, evaluating the relative contribution of the active ingredient (glyphosate) and the surfactant (polyethoxylated amine; POEA) to the genotoxicity of the commercial formulation on Anguilla anguilla. Fish were exposed to equivalent concentrations of Roundup® (58, 116 µg L⁻¹), glyphosate (17.9, 35.7 µg L⁻¹) and POEA (9.3, 18.6 µg L⁻¹), during 1 and 3 days. The comet assay was applied to blood cells, either as the standard procedure, or with an extra step involving DNA lesion-specific repair enzymes in an attempt to clarify DNA damaging mechanisms. The results confirmed the genotoxicity of Roundup®, also demonstrating the genotoxic potential of glyphosate and POEA individually. Though both components contributed to the overall genotoxicity of the pesticide formulation, the sum of their individual effects was never observed, pointing out an antagonistic interaction. Although POEA is far from being considered biologically inert, it did not increase the risk associated to glyphosate when the two were combined. The analysis of oxidatively induced breaks suggested that oxidation of DNA bases was not a dominant mechanism of damage. The present findings highlighted the risk posed to fish populations by the assessed chemicals, jointly or individually, emphasizing the need to define regulatory thresholds for all the formulation components and recommending, in particular, the revision of the hazard classification of POEA.

Sildenafil restores cognitive function without affecting ß-amyloid burden in a mouse model of Alzheimer's disease.

BACKGROUND AND PURPOSE: Inhibitors of phosphodiesterase 5 (PDE5) affect signalling pathways by elevating cGMP, which is a second messenger involved in processes of neuroplasticity. In the present study, the effects of the PDE5 inhibitor, sildenafil, on the pathological features of Alzheimer's disease and on memory-related behaviour were investigated. EXPERIMENTAL APPROACH: Sildenafil was administered to the Tg2576 transgenic mouse model of Alzheimer's disease and to age-matched negative littermates (controls). Memory function was analysed using the Morris water maze test and fear conditioning tasks. Biochemical analyses were performed in brain lysates from animals treated with saline or with sildenafil. KEY RESULTS: Treatment of aged Tg2576 animals with sildenafil completely reversed their cognitive impairment. Such changes were accompanied in the hippocampus by a reduction of tau hyperphosphorylation and a decrease in the activity of glycogen synthase kinase 3ß (GSK3ß) and of cyclin-dependent kinase 5 (CDK5) (p25/p35 ratio). Moreover, sildenafil also increased levels of brain-derived neurotrophic factor (BDNF) and the activity-regulated cytoskeletal-associated protein (Arc) in the hippocampus without any detectable modification of brain amyloid burden. CONCLUSIONS AND IMPLICATIONS: Sildenafil improved cognitive functions in Tg2576 mice and the effect was not related to changes in the amyloid burden. These data further strengthen the potential of sildenafil as a therapeutic agent for Alzheimer's disease.