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.

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.

Decrease in oxidative stress and histological changes induced by physical exercise calibrated in rats with osteoarthritis induced by monosodium iodoacetate.

OBJECTIVE: The purpose of this study was to evaluate the effects of impact exercise on the joint cartilage of rats with osteoarthritis (OA) induced by monosodium iodoacetate (MIA). METHODS: Eighteen male rats were divided into three groups of six animals each: control, OA, and OA plus exercise (OAE). The OAE group trained on a treadmill for 8 weeks. Afterward, the right joints of the animals were washed with saline solution and joint lavage was used for biochemical analyses of myeloperoxidase (MPO) and enzyme superoxide dismutase (SOD) activities and total thiol content. The same limb provided samples of the articular capsule for analyses of MPO activity and total thiol content. The left joint was used for histological analysis. RESULTS: Our results indicate that MPO activity was increased in both OA groups in the lavage as well as the articular capsule, regardless of exercise status. SOD activity was increased in animals with OA, especially in the animals that had run on the treadmill. On the other hand, thiol content in the articular capsule and joint lavage decreased in the OA group, while the OAE group had values similar to those of the control group. The histological data indicate that animals that were submitted to running exercise showed a higher preservation rate of proteoglycan content in the superficial and intermediate areas of the joint cartilage. CONCLUSION: Our results show that physical training contributes to the preservation of joint cartilage in animals with OA and to increase the defense mechanism against oxidative stress.

Intracerebral amyloidoma: case report and review of the literature.

Intracerebral amyloidoma (ICA) is a type of monoclonal immunoglobulin deposition disease (MIDD) which is accompanied by an overexpression and fibrillary assembly of monoclonal light chains, ultimately leading to nodular deposits of light chains in the form of amyloid light chain (AL-amyloid). The diagnosis is made by the histological demonstration of intracerebral masses harboring the classical staining and birefringence features of amyloid. We aim to report a case of ICA and review histological features of previous cases. A 51-year-old man with epilepsy and cognitive decline was admitted for epileptic seizures. A brain magnetic resonance imaging (MRI) disclosed periventricular enhancing lesions, hypointense on T1 and heterogeneous on T2-weighted images. A brain stereotactic biopsy was performed. The neuropathological examination revealed several congophilic nodules, allowing the diagnosis of ICA. The immunohistochemical study was positive for transthyretin (TTR), and both lambda and kappa immunoglobulin light chains. No inflammatory infiltrates were seen. Although a plasma cell clone may play a major role in the etiopathogeny of ICA, plasma cells were scarce or even absent when reviewing histological reports. ICA has a poorly understood patgogenesis. ICA may simulate malignant neoplasms, hence the need for a definite histological diagnosis.

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.

Papillary glioneuronal tumor--prognostic value of the extension of surgical resection.

OBJECTIVE: Papillary glioneuronal tumors (PGNT) is well-recognized in the literature, although reports usually have not attempted a critical analysis of their characteristics. We report two PGNT and perform a comprehensive review of the published cases, aiming to clarify their clinical, imaging and histopathological features. MATERIAL AND METHODS: We have reviewed all glioneuronal tumors diagnosed in our laboratory over the last 10 years and found 2 cases PGNTs along with their clinical, imaging and surgical data. We have processed material for light microscopy, and for immunohistochemistry study, we have used antisera against glial fibrillary acidic protein, Olig-2C, neurofilament protein, synaptophysin and Ki-67. We searched Medline (1966 through October 2007) for original articles or previous reviews. RESULTS: Case 1, a 19-year-old girl with a left, partially cystic, occipital tumor, totally removed, with no signs of recurrence 32 months after surgery, Case 2, a 9-year-old girl with a right, cystic with a solid nodule, temporal tumor, totally removed, with no signs of recurrence 19 months after surgery. Histopathology and immunohistochemistry studies favored a diagnosis of PGNT. A survey of 38 reported PGNT cases together with our two disclosed the following typical profile: young adulthood predominance, temporal lobe location, presence of cystic components: a close association with the lateral ventricles, a few anaplastic tumors, and gross total resections were usually possible with no recurrences the extent of surgical removal being the main prognostic factor. CONCLUSIONS: Although histopathology is usually characteristic, imaging features may also be important in the presurgical evaluation of PGNTs. Gross total resections are usually possible and seem to govern prognosis. However, longer follow-up data are required.

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).

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.

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.

Determination of trans-resveratrol in grapes by pressurised liquid extraction and fast high-performance liquid chromatography.

A study has been made of the extraction of trans-resveratrol from grapes using pressurised liquids (PLE); for this, the first stage was to determine the stability of this compound during extractions at different temperatures (50, 100, 150 degrees C), with quantitative recoveries being obtained up to 150 degrees C. By employing solid-phase extraction (SPE) it was possible to retain this compound and separate it from other interfering substances present in the grape. The method developed comprises a sequential extraction of the sample adsorbed (0.5g) on a polystyrene-divinylbenzene based sorbent in the extraction chamber, first with water at 40 degrees C and 40atm of pressure (three cycles of 5min), and then with methanol at 150 degrees C and 40atm (three cycles of 5min). The trans-resveratrol content of the methanolic extract is determined by means of liquid chromatography. A rapid (5min) chromatographic method employing a monolithic column, with fluorescence detection, has been developed; for this, the conditions for detection of the compound were optimised (excitation at 310nm and emission at 403nm). The analytical parameters of the method of chromatographic analysis developed have been calculated: linear range (0.11-2.75mg/L), detection limit (0.003mg/L), quantification limit (0.004mg/L). Using this method, three varieties of grape have been analysed and the concentration of trans-resveratrol in these has been determined.

Can Nassarius reticulatus be used as a bioindicator for Hg contamination? Results from a longitudinal study of the Portuguese coastline.

The focus of this work is to explore the use of the netted whelk, Nassarius reticulatus (L.), as an indicator of mercury (Hg) contamination, by assessing the concentration of Hg in the sediments and in the whelk along the entire Portuguese coast. Total Hg concentrations ranged from below the detection limit (0.01 ng absolute mercury) up to 0.87 mg kg(-1) dry weight (dwt) in sediments and between 0.06 and 1.02 mg kg(-1) (dwt) for organisms, with no significant differences between males and females. Although organic mercury was not detected in the sediments, it represented, on average, 52% of the total Hg in the whelk tissues, and as high as 88% in some cases, suggesting mercury accumulation from dietary intake. Significant negative correlations were found between the total Hg concentrations in the sediments and the log(10) of Hg concentrations in whelk tissues males (r=-0.64; P<0.01) and females (r=-0.52; P<0.01) indicating that the species is a poor indicator of Hg contamination. Nevertheless, since the highest concentrations of organic mercury in the whelk tissues were found in the least contaminated areas, this species must be highly relevant in the trophic web, namely on the possible biomagnification of mercury. The high dietary mercury accumulation from feeding on carrion and the low bioavailability of mercury to whelks in estuarine sediments may be the basis of the mercury accumulation pattern in N. reticulatus.

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.

Determination of phenolics in cosmetic creams and similar emulsions.

A new method for the analysis of phenolics in cosmetic creams has been developed, based on a systematic study of the extractability of five phenolic compounds from such emulsions. A solid-liquid extraction using ultrasound was applied as a prior stage to the chromatographic determination of phenolics in the extracts. Three solvents, hexane, methanol and water, were used as extracting agents. These solvents permit both the de-emulsification of the creams and the extraction of phenolics. A factorial fractional experimental design was developed to analyse the influence in the extraction process of five different extraction variables: ultrasound horn, temperature, extracting volume, cycle and amplitude of ultrasounds. Graphic analysis of results revealed the variables with most influence in the extraction, as well as the interactions between the variables. Finally, the influence of the extraction time and the sample quantity were also studied. With this new method, phenolics can be extracted from silicone-based cosmetic creams in 10 min, using 50 degrees C as extraction temperature. RSDs (n=6) calculated ranged from 1.5% for ferulic acid to 6.5% for epicatechin. Recoveries of between 88.9% for gallic acid and 98.4% for caffeic acid were obtained.

Innervation of human epicardial coronary veins: immunohistochemistry and vasomotility.

OBJECTIVE: The aim was to investigate the innervation and vasomotor responses to classical and putative transmitters of the coronary venous bed. METHODS: The innervation of human epicardial coronary veins was investigated using acetylcholinesterase histochemistry and immunofluorescence staining, together with antisera against the general neuronal marker protein gene product 9.5 (PGP 9.5), the catecholamine synthesising enzyme tyrosine hydroxylase, and neuropeptides [neuropeptide Y, vasoactive intestinal peptide (VIP), substance P, and calcitonin gene related peptide (CGRP)]. The vasomotor responses to noradrenaline, acetylcholine, neuropeptide Y, substance P, human alpha calcitonin gene related peptide (alpha CGRP), and VIP were tested on isolated circular human epicardial coronary vein segments. RESULTS: A network of nerve fibres was shown in the major epicardial coronary veins by means of an antiserum to PGP 9.5. The majority of the perivascular nerve fibres possessed neuropeptide Y and tyrosine hydroxylase immunoreactivity. Only a few nerve fibres displayed substance P, CGRP, and VIP immunoreactivity and acetylcholinesterase activity. Noradrenaline and acetylcholine induced powerful contractions of all the tested segments, whereas no contraction was induced by neuropeptide Y, alpha CGRP, substance P, or VIP. All segments precontracted with U46619 responded with potent relaxation to alpha CGRP, substance P, and VIP, whereas noradrenaline and acetylcholine only in low concentrations induced weak relaxation of a few of the segments. No relaxation was induced by neuropeptide Y. CONCLUSIONS: This is the first study to demonstrate comprehensively the perivascular innervation of human coronary veins and corresponding vasomotor effects, suggesting a role in regulation of the coronary venous circulation.

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 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.

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.