Evaluation of Ki-67 expression in feline non-ocular melanocytic tumours

Fifty tumours located in skin (n = 33) and mucosae (n = 17) were included. Forty-eight percent and 95% of amelanotic tumours (n = 21) stained positive for Melan A and S100, respectively. Most achromic tumours were mucosal (P  5, Ki-67 > 20% and lack of treatment administration. On multivariable analysis, only tumour histotype and treatment retained prognostic significance. Conclusions Although the majority of feline NOMs behave aggressively, Ki-67 index, together with other parameters, may contribute to prognostic assessment. Prospective studies on homogeneous populations are warranted to identify reliable threshold values for this marker

Evidence of temperature-dependent effects on the estrogenic response of fish: implications with regard to climate change

The official published version can be obtained from the link below - Copyright @ 2008 Elsevier BV.Chemical risk assessment is fraught with difficulty due to the problem of accounting for the effects of mixtures. In addition to the uncertainty arising from chemical-to-chemical interactions, it is possible that environmental variables, such as temperature, influence the biological response to chemical challenge, acting as confounding factors in the analysis of mixture effects. Here, we investigate the effects of temperature on the response of fish to a defined mixture of estrogenic chemicals. It was anticipated that the response to the mixture may be exacerbated at higher temperatures, due to an increase in the rate of physiological processing. This is a pertinent issue in view of global climate change. Fathead minnows (Pimephales promelas) were exposed to the mixture in parallel exposure studies, which were carried out at different temperatures (20 and 30 degrees C). The estrogenic response was characterised using an established assay, involving the analysis of the egg yolk protein, vitellogenin (VTG). Patterns of VTG gene expression were also analysed using real-time QPCR. The results revealed that there was no effect of temperature on the magnitude of the VTG response after 2 weeks of chemical exposure. However, the analysis of mixture effects at two additional time points (24 h and 7 days) revealed that the response was induced more rapidly at the higher temperature. This trend was apparent from the analysis of effects both at the molecular and biochemical level. Whilst this indicates that climatic effects on water temperature are not a significant issue with regard to the long-term risk assessment of estrogenic chemicals, the relevance of short-term effects is, as yet, unclear. Furthermore, analysis of the patterns of VTG gene expression versus protein induction gives an insight into the physiological mechanisms responsible for temperature-dependent effects on the reproductive phenology of species such as roach. Hence, the data contribute to our understanding of the implications of global climate change for wild fish populations.This work was funded by a grant from the Natural Environment Research Council NE/D00389X/1). Additional support was provided by a small research grant from the Fisheries Society of the British Isles

Development of a feed additive to reduce caecal Campylobacter jejuni in broilers at slaughter age: from in vitro to in vivo, a proof of concept

Aim: In vitro and in vivo challenge studies were undertaken to develop an in-feed additive of microencapsulated propionic, sorbic acids and pure botanicals to control Campylobacter jejuni in broilers at slaughter age. Methods and Results: Organic acids (OA) and pure botanicals were tested in vitro against Camp. jejuni, whereas in vivo, chickens were fed either a control diet, or increasing doses of the additive for 42 days (experiment 1); in the second experiment, chickens received the additive at 01 or 03% from day 0 to 21 or from day 22 to 42. The additive consistently reduced Camp. jejuni caecal counts at any given dose (exp. 1) or inclusion plan (exp. 2). Moreover, it was able to reduce the number of goblet cells and modify mucin glycoconjugates biosynthesis pattern. Conclusions: We developed an additive that was effective in reducing Camp. jejuni in slaughter-age chickens even at low doses (01%). That efficacy was the result of the synergistic action between OA and botanicals. Significance and Impact of the Study: This study provides a strategy to reduce Camp. jejuni in broilers and, as a consequence, to improve the safety of the food chain. Moreover, data suggest that a treatment limited to the last weeks before slaughter would allow to save on inclusion of the additive throughout the whole production cycle

Biochemically Controlled Release of Dexamethasone Covalently Bound to PEDOT

reserved9noPEDOT (Poly(3,4-ethylenedioxythiophene)) is one of the most promising electrode materials for biomedical applications like neural recording and stimulation, thanks to its enhanced biocompatibility and electronic properties. Drug delivery by PEDOT is typically achieved by incorporating drugs as dopants during the electrodeposition procedure and a subsequent release can be promoted by applying a cathodic trigger that reduces PEDOT while enabling the drug to diffuse. This approach has several disadvantages including, for instance, the release of contaminants mainly due to PEDOT decomposition during electrochemical release. Herein we describe a new strategy based on the formation of a chemical linkage between the drug and the conductive polymer. In particular, dexamethasone was successfully integrated into a new electropolymerized PEDOT–Dex composite, leading to a self-adjusting drug release system based on a biochemically hydrolysable bond between dexamethasone and PEDOT.mixedCarli S.; Trapella C.; Armirotti A.; Fantinati A.; Ottonello G.; Scarpellini A.; Prato M.; Fadiga L.; Ricci D.Carli, S.; Trapella, C.; Armirotti, A.; Fantinati, A.; Ottonello, G.; Scarpellini, A.; Prato, M.; Fadiga, L.; Ricci, D

Effect of the novel synthetic cannabinoids AKB48 and 5F-AKB48 on “tetrad”, sensorimotor, neurological and neurochemical responses in mice. In vitro and in vivo pharmacological studies

Rationale: AKB48 and its fluorinate derivate 5F-AKB48 are two novel synthetic cannabinoids belonging to a structural class with an indazole core structure. They are marketed as incense, herbal preparations or chemical supply for their psychoactive Cannabis-like effects. Objectives: The present study was aimed at investigating the in vitro and in vivo pharmacological activity of AKB48 and 5F-AKB48 in male CD-1 mice and comparing their in vivo effects with those caused by the administration of Δ9-THC and JWH-018. Results: In vitro competition binding experiments performed on mouse and human CB1 and CB2 receptors revealed a nanomolar affinity and potency of the AKB48 and 5F-AKB48. In vivo studies showed that AKB48 and 5F-AKB48, induced hypothermia, increased pain threshold to both noxious mechanical and thermal stimuli, caused catalepsy, reduced motor activity, impaired sensorimotor responses (visual, acoustic and tactile), caused seizures, myoclonia, hyperreflexia and promoted aggressiveness in mice. Moreover, microdialysis study in freely moving mice showed that systemic administration of AKB48 and 5F-AKB48 stimulated dopamine release in the nucleus accumbens. Behavioural, neurological and neurochemical effects were fully prevented by the selective CB1 receptor antagonist/inverse agonist AM 251. Conclusions: For the first time, the present study demonstrates the overall pharmacological effects induced by the administration of AKB48 and 5F-AKB48 in mice and suggests that the fluorination can increase the power and/or effectiveness of SCBs. Furthermore, this study outlines the potential detrimental effects of SCBs on human health

Metabolic profile and pharmaco-toxicological effects of MPA in mouse model

Methiopropamine (MPA) is a thiophene ring-based structural analog of methamphetamine that is categorized as a novel psychoactive substance. MPA acts primary as a norepinephrine-dopamine reuptake inhibitor and, secondary, as a serotonin reuptake inhibitor. In human, MPA induces stimulation, alertness and increase of focus and energy. However, important side-effects were reported after MPA administration such as tachycardia, anxiety, panic attacks, perspiration, headache and difficulty in breathing. To date, few data are available on the pharmaco-toxicological effects of MPA and on its metabolism in vivo. To this purpose we investigated the acute in vivo effects induced by MPA on visual, acoustic and tactile sensorimotor responses, body temperature, pain threshold, motor activity, cardiovascular and cardiorespiratory changes in CD-1 male mice. In vitro (from human liver microsomes preparation) and in vivo (from CD-1 male mice urine) metabolism studies were also conducted in order to characterize the phase I metabolic profile of MPA, with the aim to select both the best marker(s) of intake and the responsible of the potential behavioural effects. Systemic administration of MPA (0.01-30 mg/kg) impaired visual placing response, facilitated acoustic and tactile response, induced hypothermia, increased mechanical and thermal analgesia stimulated locomotor activity and induced motor stereotypies in mice. MPA strongly affected cardiovascular and respiratory parameters. It induced tachycardia, increased diastolic and systolic blood pressure, caused vasoconstriction. Moreover, MPA increased breath rate but it reduced SpO2 saturation. The acute administration of MPA at 10 and 30 mg/kg caused the death (~30% and ~40%, respectively) of mice. Metabolism studies show that MPA is exensively oxidated to form mainly nor-MPA, hydroxy-MPA, oxo-MPA. The best markers of intake and the compounds detected in huge amount during the behavioural tests are the MPA itself and its hydroxylated and demethylated metabolites. The experimental evidence obtained in this study demonstrates for the first time that MPA impairs sensorimotor responses, has psychostimulant effect, causes cardiovascular and respiratory alterations, thus suggesting its possible hazard for human health. This research has been funded by the Drug Policies Department, Presidency of the Council of Ministers, Italy (project: “Effects of NPS: development of a multicentric research for the information enhancement of the Early Warning System” to MM) and FIRB 2012 to FDG

Conjugation of LasR Quorum-Sensing Inhibitors with Ciprofloxacin Decreases the Antibiotic Tolerance of P. aeruginosa Clinical Strains

Pseudomonas aeruginosa is a Gram-negative bacterium that commonly infects subjects with weakened immune system causing deadly infections above all at pulmonary level. During infection, P. aeruginosa produces a well-organized bacterial structure, called biofilm, activating the quorum-sensing (QS) signaling, a mechanism of gene regulation. In this work, we synthesized already known QS inhibitors (QSi) designed on the scaffold of the N-(3-oxododecanoyl) homoserine lactone (3O-C12-HSL) QS molecule and conjugated them with ciprofloxacin to inhibit P. aeruginosa biofilm formation and increase the antibiotic susceptibility of clinical strains. We identified, for the first time, a QSi conjugated with ciprofloxacin (ET37), that is able to reduce the formation of biofilm and the onset of tolerant clones in P. aeruginosa clinical strains. This compound could have a wide application in clinical setting. The possibility to affect biofilm formation in chronically infected patients, such as patients affected by cystic fibrosis, and to reduce the onset of ciprofloxacin resistance would improve patient healing and allow to decrease antibiotic drug dosage