Search

Preclinical testing of novel therapeutics for chronic hepatitis B (CHB) requires suitable animal models. Equids host homologs of hepatitis C virus (HCV). Because coinfections of hepatitis B virus (HBV) and HCV occur in humans, we screened 2,917 specimens from equids from five continents for HBV. We discovered a distinct HBV species (Equid HBV, EqHBV) in 3.2% of donkeys and zebras by PCR and antibodies against EqHBV in 5.4% of donkeys and zebras. Molecular, histopathological, and biochemical analyses revealed that infection patterns of EqHBV resembled those of HBV in humans, including hepatotropism, moderate liver damage, evolutionary stasis, and potential horizontal virus transmission. Naturally infected donkeys showed chronic infections resembling CHB with high viral loads of up to 2.6 × 10(9) mean copies per milliliter serum for >6 mo and weak antibody responses. Antibodies against Equid HCV were codetected in 26.5% of donkeys seropositive for EqHBV, corroborating susceptibility to both hepatitis viruses. Deltavirus pseudotypes carrying EqHBV surface proteins were unable to infect human cells via the HBV receptor NTCP (Na(+)/taurocholate cotransporting polypeptide), suggesting alternative viral entry mechanisms. Both HBV and EqHBV deltavirus pseudotypes infected primary horse hepatocytes in vitro, supporting a broad host range for EqHBV among equids and suggesting that horses might be suitable for EqHBV and HBV infections in vivo. Evolutionary analyses suggested that EqHBV originated in Africa several thousand years ago, commensurate with the domestication of donkeys. In sum, EqHBV naturally infects diverse equids and mimics HBV infection patterns. Equids provide a unique opportunity for preclinical testing of novel therapeutics for CHB and to investigate HBV/HCV interplay upon coinfection.

Background. The primary somatosensory cortex (S1), once thought to be only modality-specific, is involved in higher level functions like emotion recognition and motor learning by observation. S1’ functions may extend even to memory: some theories speculate that these areas may be involved not only in the perception of intrinsic features of the percept but also in the retention of that information. In a recent EEG study, Galvez-Pol suggested that the neural responses of somatosensory cortices to visually perceived body-related information probably reflected their involvement in short-term memory (STM). Aim. The present experiment looks for causal evidence that S1 is implicated in the retention of visual information that may be salient for this cortical area 1. To this purpose we interfered with S1 activity by means of repetitive Transcranial Magnetic Stimulation (rTMS) during a STM task where body-related information had to be retained. Methods. Eighteen healthy volunteers took part in a three sessions within-subjects experiment. The STM task (i.e., delayed match-to-sample task) consisted in the rapid presentation of two lateralized arrays, each one depicting three body-related stimuli (i.e., pictures of hands in different position, taken from previous literature. In the task, half of the trials were identical and half were different for only one stimulus. In each session, every subject performed the task with and without rTMS, with the latter condition serving as a baseline. The rTMS protocol consisted in a train of 3 TMS-pulses (at 10 Hz frequency, fixed intensity of 60% of the maximum stimulator output) delivered after 200 ms from the offset of the first array (i.e. during the retention phase). As control sites for rTMS effects, we stimulated two other cortical areas: the dorsolateral prefrontal cortex (dlPFC; an area commonly activated during working memory tasks) and the lateral occipital cortex (lOC; an area activated in feature-based analysis of visual stimuli). Only the right hemisphere was targeted. Results. We analyzed the sensitivity detection index (d') in the STM task through a 2 X 3 repetitive-measures ANOVA with the within-subjects factors “Condition” (baseline/rTMS) and “Area” (S1/dlPFC/lOC). We found a significant interaction between these two factors (p=0.036). Planned comparisons revealed an improvement of subjects’ performance in the STM task only when rTMS is delivered over S1 (vs. baseline, p

Background & object. The primary somatosensory cortex (S1), once thought to be only modality-specific, is involved in higher level functions like emotion recognition and motor learning by observation. S1 functions may extend even to memory: some theories speculate that sensorimotor areas may be involved not only in the perception of intrinsic features of the percept but also in the retention of that information. In a recent EEG study, it has been suggested that the neural responses of somatosensory cortices to visually perceived body-related information probably reflect their involvement in short-term memory (STM). The present experiment looks for causal evidence that S1 is implicated in the retention of visual information that may be salient for this cortical area1. To this purpose we interfered with S1 activity by means of repetitive Transcranial Magnetic Stimulation (rTMS) during a STM task where body-related information had to be retained. Methods & materials. Eighteen healthy volunteers took part in a three sessions within-subjects experiment. The STM task (i.e., delayed match-to-sample task) consisted in the rapid presentation of two lateralized arrays, each one depicting three body-related stimuli (i.e., pictures of hands in different position, taken from previous literature2). In the task, half of the trials were identical and half were different for only one stimulus. In each session, every subject performed the task with and without rTMS, with the latter condition serving as a baseline. The rTMS protocol consisted in a train of 3 TMS-pulses (at 10 Hz frequency, fixed intensity of 60% of the maximum stimulator output) delivered after 200 ms from the offset of the first array (i.e. during the retention phase). As control sites for rTMS effects, we stimulated two other cortical areas: the dorsolateral prefrontal cortex (dlPFC; an area commonly activated during WM tasks) and the lateral occipital cortex (lOC; an area activated in feature-based analysis of visual stimuli). Only the right hemisphere was targeted. Results. We analyzed the sensitivity detection index (d') in the STM task through a 2 X 3 repetitive-measures ANOVA with the within-subjects factors “Condition” (baseline vs. rTMS) and “Area” (S1, dlPFC, lOC). We found a statistically significant interaction between these two factors (p=0.036). Planned comparisons revealed an improvement of subjects’ performance in the STM task only when rTMS is delivered over S1 (vs. baseline, p

In Italy despite the increasing use of donkeys in leisure activities, onotherapy and milk production, there are no epidemiological data regarding the parasites affecting this species. A survey was conducted to determine the prevalence of the principal endoparasitic infections in 50 donkey farms. Between March 2012-2015, faecal samples were collected from 961 donkeys and examined individually using a modified McMaster technique. In addition a centrifugation/flotation technique (Proudman test) and a sedimentation technique were used for the diagnosis of Anoplocephalidae and Fasciola hepatica eggs, respectively. On each farm pooled coprocoltures were made. Strongyles were the most common parasitic species found, with a prevalence of 88.5%. 11.4% of donkeys were not infected and infection intensity was lower than the donkey cut-off selective therapy of 300 eggs per gram (epg) in 33.4% of animals. The other epg values were 300–500 in 14.3%, 501–1,000 in 26.0%, 1,001-2,000 in 12.8% and > 2,000 in 2.1% of donkeys. In all tested farms, coprocoltures revealed the presence of Cyathostomes (100%) followed by Strongylus vulgaris in 13 farms (26%), Poteriostomum spp. (24%), Triodontophorus spp. (6%) and S. edentatus (4%). Other parasites found in studied donkeys were: Oxyuris equi (7.8%), Dictyocaulus arnfieldi (4.4%), Parascaris equorum (2.8%), Anoplocephala perfoliata (1.1%) Eimeria leuckarti (0.6%) and Strongyloides westeri (0.1%). No positivity was detected for F. hepatica. This study represents the first large-scale survey in Italy and added data to the parasitological scenario of donkey population.

Twenty-eight buffalo cows were used to evaluate the effect of housing system on a range of behavioural and physiological variables from February to July. Fourteen cows were group-housed in a loose open-sided barn with a concrete floor and 10 m2/head as space allowance (Group IS). Fourteen others were group-housed in a similar barn but they could also benefit of an outdoor yard with 500 m2/head as space allowance, spontaneous vegetation and free access to potholes for wallowing (Group TS). Six behavioural recordings were performed using instantaneous scan sampling. Data were expressed as proportion of subjects observed in each behavioural category. Phytoemagglutinin (PHA) and ovalbumin were used to assess cellular and humoral immune responses, respectively. Cortisol concentration was evaluated prior to and after exogenous ACTH injection. The metabolic status of the animals and milk production were also monitored. Immune responses, metabolite concentrations and milk production were not affected by treatment. The provision of an housing system close to natural conditions was able to improve the welfare of buffalo cows as indicated by the expression of some species-specific natural behaviours (i.e. grazing and wallowing) and the reduction of idling (P

Objective Social cognition and executive deficits are frequent in neurological and neuropsychiatric disorders. Yet, there is a paucity of standardized domain-specific psychometric tools for the assessment of complex decision-making, social cognition (i.e., Theory of Mind), and empathy. To this aim, this study intended at providing normative data in an Italian population sample for the Iowa Gambling Task, the Reading the Mind in the Eyes Test, and the Interpersonal Reactivity Index. Method The Iowa Gambling Task, the Reading the Mind in the Eyes Test, and the Interpersonal Reactivity Index were administered to 462 healthy Italian participants aged between 18 and 91 years, considering demographic factors. A confirmatory factor analysis was run to test for unidimensionality. Normative values were derived by means of the Equivalent Scores method. Results Education predicted all tests’ scores, age influenced scores at the Interpersonal Reactivity Index and the Reading the Mind in the Eyes Test, gender predicted empathic abilities only. The three tests did not underpin a unidimensional structure. Conclusions The present work provides demographically adjusted Italian normative data for a set of tests assessing real-life decision-making, emotion recognition and empathy, filling a gap within II-level, domain-specific, neuropsychological testing. For the first time, the Interpersonal Reactivity Index is proposed as an adjuvant neuropsychological tool, while the standardization of the Iowa Gambling Task offers a new scoring system for advantageous/disadvantageous choices.

Starting from the early 2000s, paired associative stimulation (PAS) protocols have been used in humans to study brain connectivity in motor and sensory networks by exploiting the intrinsic properties of timing-dependent cortical plasticity. In the last 10 years, PAS have also been developed to investigate the plastic properties of complex cerebral systems, such as the frontal ones, with promising results. In the present work, we review the most recent advances of this technique, focusing on protocols targeting frontal cortices to investigate connectivity and its plastic properties, subtending high-order cognitive functions like memory, decision-making, attentional, or emotional processing. Overall, current evidence reveals that PAS can be effectively used to assess, enhance or depress physiological connectivity within frontal networks in a timing-dependent way, in turn modulating cognitive processing in healthy and pathological conditions.

In the early 2000s, a novel non-invasive brain stimulation protocol, the paired associative stimulation (PAS), was introduced, allowing to induce and investigate Hebbian associative plasticity within the humans' motor system, with patterns resembling spike-timing-dependent plasticity properties found in cellular models. Since this evidence, PAS efficacy has been proved in healthy, and to a lesser extent, in clinical populations. Recently, novel 'modified' protocols targeting sensorimotor and crossmodal networks appeared in the literature. In the present work, we have reviewed recent advances using these 'modified' PAS protocols targeting sensory and motor cortical networks. To better categorize them, we propose a novel classification according to the nature of the peripheral and cortical stimulations (i.e., within-system, cross-systems, and cortico-cortical PAS). For each protocol of the categories mentioned above, we describe and discuss their main features, how they have been used to study and promote brain plasticity, and their advantages and disadvantages. Overall, current evidence suggests that these novel non-invasive brain stimulation protocols represent very promising tools to study the plastic properties of humans' sensorimotor and crossmodal networks, both in the healthy and in the damaged central nervous system.

INTRODUCTION. Horses are naturally parasitized by several genus/group of helminths and intestinal strongyles are the most common. Presently three anthelmintic classes are registered in horses: tetrahydropyrimidines, benzimidazoles and macrocyclic lactones (Gokbulut and McKellar, 2018 Vet Parasitol. 261: 27-52). The traditional approach to helminths control is based on interval dose program without a diagnosis, furthermore the frequent and reason less drug use has led to development of anthelmintic resistance (Leathwick et al., 2019 Vet Parasitol. 209: 210-20). Equine deworming scheme for adult horses is based on strongyle Fecal Egg Count (FEC) and on treatment of horses with FEC>200 egg per gram of feces (EPG) (Selective Treatment) (AAEP Internal Parasite Control Guidelines, 2019). FEC is also crucial to evaluate the treatment efficacy. The aims of this work were to carry out a survey on intestinal strongyles in horses in Italy and to propose a veterinary assistance considering a Parasitological Assistance Program in Equids (PAPE) based on coprological diagnosis (FEC). MATERIALS AND METHODS. The study was performed on 7,796 horses bred in 542 Italian farms. Individual FECs were performed using Mini-FLOTAC technique and a Sheather's sugar solution with a specific gravity of 1.250. Individual coprocultures were set up for each horse with FEC>200EPG. RESULTS AND CONCLUSIONS. Intestinal strongyle eggs were found in 420 (77.5%) tested farms with a prevalence at animal level of 57.6% (4,492/7,796). The average EPG was 441.2±622.0 (min 5 max 7,100). Regarding the egg shedding level, 71.4% of horses had a FEC 500 EPG (high contaminators). Coprocultures revealed L3 of Cyathostominae (100%), Poteriostomum (3.2%), Gyalocephalus (2.1%), Strongylus vulgaris (1.9%), Strongylus edentates (1.1%), Triodontophorus (0.7%) and Oesophagodontus (0.2%). Considering that FEC was

Donkeys have been used as working animals for transport and farm activities worldwide. Recently, in European countries, there has been an increasing interest in donkeys due to their use as pets, onotherapy or milk production. During 2014–2016, a countrywide survey was conducted to determine prevalence and risk factors of principal helminth infections in 1775 donkeys in 77 Italian farms. A questionnaire on management and parasite control practices was filled out for each farm. Faecal samples were examined using a modified McMaster technique, a centrifugation/flotation method and a sedimentation technique. Pooled coprocultures were performed for differentiation of strongylid eggs. Strongyles were the most common parasites detected (84.9%), followed by Dictyocaulus arnfieldi (6.9%), Oxyuris equi (5.8%), Parascaris spp. (3.6%), Anoplocephala spp. (1.0%), Strongyloides westeri (0.3%). Coprocultures revealed an omnipresence of cyathostomins (100%), followed by Strongylus vulgaris (31.0%), Poteriostomum spp. (25.0%), Triodontophorus spp. (9.0%), Strongylus edentatus (7.0%), Strongylus equinus (5.0%). Logistic regression analysis identified breed, co-pasture with horses, living area, herd size and number of treatments as significantly associated with strongyles. Sex, age, living area and herd size were significantly associated with Parascaris spp. Dictyocaulus arnfieldi was significantly associated with sex, grass, co-pasture with horses, living area and herd size. Strongylus vulgaris was significantly associated with living area and herd size. The mean number of anthelmintic treatments/year was 1.4; most of the donkeys (71.8%) were dewormed using an ivermectin drug. It is important to design parasite programs to specifically address both D. arnfieldi and S. vulgaris in donkeys, and this is especially important if donkeys co-graze with horses.

Intestinal strongyles are the most common endoparasites of horses, and anthelmintic treatments are the main strategy to control these nematodes. However, the development of anthelmintic resistance has led to a decreased efficacy of synthetic drugs, and for this reason, there is a growing interest in alternative control strategies as the use of medicinal plants. The aim of the present study was to determine the in vivo efficacy of garlic (Allium sativum) in horses naturally infected by intestinal strongyles. The field trial was conducted in a horse trotter farm in Southern Italy. Fifteen mares were selected based on fecal egg count >200 eggs per gram and allocated into three groups of five animals: fresh garlic group (FG group), animals received 40 g of fresh crushed garlic once daily for 15 days; dry garlic group (DG group), animals received 40 g of commercial dry garlic flakes food supplement once daily for 15 days; and control group (C group), not treated. Two weeks after the first administration of garlic, fecal egg count reduction test showed failure of garlic to reduce intestinal strongyles egg shedding (−11.7% and −19.4% for FG and DG groups, respectively). Red blood cell count values were in the normal ranges over the entire period of garlic administration. In our study model, the oral administration of garlic formulations has no effect on reducing the egg shedding of intestinal strongyles, and the garlic supplementation over a short period of time is not responsible for hematological changes in horses.

Mebendazole (MBZ) has been licensed for use in horses and donkeys, however there are no data available in the literature regarding its pharmacokinetic disposition and efficacy in donkeys. This study was designed to determine the plasma disposition, milk excretion and anthelmintic efficacy of MBZ in donkeys naturally infected by Cyathostominae. The animals were allocated to three groups, each of six donkeys. One group was untreated control (C-group) and the others were treated using a paste formulation of MBZ administered per os at the manufacturer's recommended horse dosage of 10 mg/kg body weight (MBZ 1) and at the double horse dosage 20 mg/kg body weight (MBZ 2). Blood and milk samples were collected at various times between 1 h and 120 h post treatment and analyzed by high performance liquid chromatography with photodiode array detector. Individual FECs (Faecal Egg Counts) were performed on each animal before the treatment (day-3) and weekly from day 7 until day 56 post treatment using a modified McMaster technique. The plasma concentrations and systemic exposure of MBZ in donkeys were relatively lower compared with the other methylcarbamate benzimidazoles. Dose-dependent plasma dispositions of MBZ were observed at the increased dosage (10 mg/kg vs 20 mg/kg) in donkeys. MBZ was not detected in any milk samples at a dosage of 10 mg/kg. However, the parent drug reached 0.01 μg/ml peak milk concentration at 10.66 h and AUCmilk/AUCplasma value was 0.18 ± 0.02 at a dosage of 20 mg/kg bodyweight. This study indicated that per os administration of MBZ has a minimal disposition rate into the milk and may be used in lactating donkeys with zero milk-withdrawal period. The results of FECRT for both MBZ dosages were efficient (>95% efficacy) until day 28. This trial demonstrates that MBZ oral paste at horse dosage (10 mg/kg B.W.) was effective and safety for the treatment of Cyathostominae in donkeys. Therefore, similar dosage regimens of MBZ could be used for horses and donkeys.

Collective evidence indicates that motor neuron degeneration in amyotrophic lateral sclerosis (ALS) is non-cell-autonomous and requires the interaction with the neighboring astrocytes. Recently, we reported that a subpopulation of spinal cord astrocytes degenerates in the microenvironment of motor neurons in the hSOD1 G93A mouse model of ALS. Mechanistic studies in vitro identified a role for the excitatory amino acid glutamate in the gliodegenerative process via the activation of its inositol 1,4,5-triphosphate (IP 3)-generating metabotropic receptor 5 (mGluR5). Since non-physiological formation of IP 3 can prompt IP 3 receptor (IP 3R)-mediated Ca 2+ release from the intracellular stores and trigger various forms of cell death, here we investigated the intracellular Ca 2+ signaling that occurs downstream of mGluR5 in hSOD1 G93A-expressing astrocytes. Contrary to wild-type cells, stimulation of mGluR5 causes aberrant and persistent elevations of intracellular Ca 2+ concentrations ([Ca 2+] i) in the absence of spontaneous oscillations. The interaction of IP 3Rs with the anti-apoptotic protein Bcl-X L was previously described to prevent cell death by modulating intracellular Ca 2+ signals. In mutant SOD1-expressing astrocytes, we found that the sole BH4 domain of Bcl-X L, fused to the protein transduction domain of the HIV-1 TAT protein (TAT-BH4), is sufficient to restore sustained Ca 2+ oscillations and cell death resistance. Furthermore, chronic treatment of hSOD1 G93A mice with the TAT-BH4 peptide reduces focal degeneration of astrocytes, slightly delays the onset of the disease and improves both motor performance and animal lifespan. Our results point at TAT-BH4 as a novel glioprotective agent with a therapeutic potential for ALS. © The Author 2011. Published by Oxford University Press. All rights reserved