Intracerebral administration of Interleukin-12 (IL-12) and IL-18 modifies the course of mouse scrapie

BACKGROUND: Prion diseases are characterised by a neurodegenerative pattern in which the function of immune system remains still elusive. In the present study, we evaluate if an exogenous treatment with Interleukin-12 (IL-12) and IL-18, able to activate microglia, is able to affect scrapie pathogenesis. RESULTS: Cytokines injected intracranially, induced a strong inflammatory response characterised by TNF-α production and microglia activation. Two groups of mice were injected intracerebrally with high dose of ME7 strain of scrapie containing IL-12 and IL-18 or sterile saline. Cytokines-treated mice showed a more pronounced accumulation of PrP(Sc )in brain tissues at 90 days post-inoculation and a shorter mean survival times than untreated mice. CONCLUSION: We can conclude that intracerebral administration of IL-12 and IL-18 can modulate scrapie pathogenesis possibly through a microglia-mediated pattern

Integrated analytical approach in veal calvesadministered the anabolic androgenic steroidsboldenone and boldione: urine and plasma kineticprofile and changes in plasma protein expression

Surveillance of illegal use of steroids hormones in cattle breeding is a key issue to preserve human health. To this purpose, an integrated approach has been developed for the analysis of plasma and urine from calves treated orally with a single dose of a combination of the androgenic steroids boldenone and boldione. A quantitative estimation of steroid hormones was obtained by LC-APCI-QMS/ MS analysis of plasma and urine samples obtained at various times up to 36 and 24 h after treatment, respectively. These experiments demonstrated that boldione was never found, while boldenone a- and b-epimers were detected in plasma and urine only within 2 and 24 h after drug administration, respectively. Parallel proteomic analysis of plasma samples was obtained by combined 2-DE,MALDI-TOF-MS and mLC-ESI-IT-MS/MS procedures. A specific protein, poorly represented in normal plasmasamples collected before treatment,was found upregulated even 36 h after hormone treatment.Extensivemassmapping experiments proved this component as an N-terminal truncated form of apolipoprotein A1 (ApoA1), a protein involved in cholesterol transport. The expression profile of ApoA1 analysed byWestern blot analysis confirmed a significant and time dependent increase of thisApoA1 fragment. Then, provided that further experiments performed with a growth-promoting schedule will confirm these preliminary findings, truncated ApoA1 may be proposed as a candidate biomarker for steroid boldenone and possibly other anabolic androgens misuse in cattle veal calves, when no traces of hormones are detectable in plasma or urine

Efficient Transmission and Characterization of Creutzfeldt–Jakob Disease Strains in Bank Voles

Transmission of prions between species is limited by the “species barrier,” which hampers a full characterization of human prion strains in the mouse model. We report that the efficiency of primary transmission of prions from Creutzfeldt–Jakob disease patients to a wild rodent species, the bank vole (Clethrionomys glareolus), is comparable to that reported in transgenic mice carrying human prion protein, in spite of a low prion protein–sequence homology between man and vole. Voles infected with sporadic and genetic Creutzfeldt–Jakob disease isolates show strain-specific patterns of spongiform degeneration and pathological prion protein–deposition, and accumulate protease-resistant prion protein with biochemical properties similar to the human counterpart. Adaptation of genetic Creutzfeldt–Jakob disease isolates to voles shows little or no evidence of a transmission barrier, in contrast to the striking barriers observed during transmission of mouse, hamster, and sheep prions to voles. Our results imply that in voles there is no clear relationship between the degree of homology of the prion protein of the donor and recipient species and susceptibility, consistent with the view that the prion strain gives a major contribution to the species barrier. The vole is therefore a valuable model to study human prion diversity and, being susceptible to a range of animal prions, represents a unique tool for comparing isolates from different species

PLEKHG5 deficiency leads to an intermediate form of autosomal-recessive Charcot-Marie-Tooth disease

Charcot-Marie-Tooth disease (CMT) comprises a clinically and genetically heterogeneous group of peripheral neuropathies characterized by progressive distal muscle weakness and atrophy, foot deformities and distal sensory loss. Following the analysis of two consanguineous families affected by a medium to late-onset recessive form of intermediate CMT, we identified overlapping regions of homozygosity on chromosome 1p36 with a combined maximum LOD score of 5.4. Molecular investigation of the genes from this region allowed identification of two homozygous mutations in PLEKHG5 that produce premature stop codons and are predicted to result in functional null alleles. Analysis of Plekhg5 in the mouse revealed that this gene is expressed in neurons and glial cells of the peripheral nervous system, and that knockout mice display reduced nerve conduction velocities that are comparable with those of affected individuals from both families. Interestingly, a homozygous PLEKHG5 missense mutation was previously reported in a recessive form of severe childhood onset lower motor neuron disease (LMND) leading to loss of the ability to walk and need for respiratory assistance. Together, these observations indicate that different mutations in PLEKHG5 lead to clinically diverse outcomes (intermediate CMT or LMND) affecting the function of neurons and glial cell

Correlation between infectivity and disease associated prion protein in the nervous system and selected edible tissues of naturally affected scrapie sheep

<div><p>The transmissible spongiform encephalopathies (TSEs) or prion diseases are a group of fatal neurodegenerative disorders characterised by the accumulation of a pathological form of a host protein known as prion protein (PrP). The validation of abnormal PrP detection techniques is fundamental to allow the use of high-throughput laboratory based tests, avoiding the limitations of bioassays. We used scrapie, a prototype TSE, to examine the relationship between infectivity and laboratory based diagnostic tools. The data may help to optimise strategies to prevent exposure of humans to small ruminant TSE material via the food chain. Abnormal PrP distribution/accumulation was assessed by immunohistochemistry (IHC), Western blot (WB) and ELISA in samples from four animals. In addition, infectivity was detected using a sensitive bank vole bioassay with selected samples from two of the four sheep and protein misfolding cyclic amplification using bank vole brain as substrate (vPMCA) was also carried out in selected samples from one animal. Lymph nodes, oculomotor muscles, sciatic nerve and kidney were positive by IHC, WB and ELISA, although at levels 100–1000 fold lower than the brain, and contained detectable infectivity by bioassay. Tissues not infectious by bioassay were also negative by all laboratory tests including PMCA. Although discrepancies were observed in tissues with very low levels of abnormal PrP, there was an overall good correlation between IHC, WB, ELISA and bioassay results. Most importantly, there was a good correlation between the detection of abnormal PrP in tissues using laboratory tests and the levels of infectivity even when the titre was low. These findings provide useful information for risk modellers and represent a first step toward the validation of laboratory tests used to quantify prion infectivity, which would greatly aid TSE risk assessment policies.</p></div

Grass intake and meat oxidative status of geese reared in three different agroforestry systems

The agroforestry system could be considered a dynamic management of the natural resource based on the integration of trees with crops or livestock. The purpose of this study was to evaluate the grass intake and the oxidative status of meat of geese reared in three different agroforestry systems: apple orchard (AO), olives trees (OT) and vineyard (V). Eighty one-day old Romagnola geese of both sexes were divided in four homogeneous groups: control (C), with indoor density of 5 geese/m2 and without pasture access, and the three agroforestry systems (AO, OT, V), with 1 hectare of pasture each. The geese were reared inside a poultry house until 20 days of age. At 21 days of age the animals belonging to AO, OT and V were allowed to outdoor access (pasture), whereas geese of the C group were kept indoor. At 150 days of age, the geese were slaughtered in a commercial slaughterhouse. After 24 h of storage at +4°C the breast and drumstick muscles were analysed to determine the fatty acid profile, the antioxidants content and the oxidative status. All the data were statistically analysed with ANOVA. The results showed that the grazing activity of geese improved the n-3 polyunsaturated fatty acids content, the n-6/n-3 ratio and the antioxidant content, especially in geese kept in the agroforestry systems enriched with trees (AO and OT). Indeed, the presence of trees make animals feel protected and stimulated them to explore the pasture and consequently to consume more grass. However, the best oxidative status was exhibited by the C geese. In the other groups the higher antioxidants intake through grass was not able to counteract the higher oxidative thrust and consequently, the meat of outdoor reared geese was characterized by a worst oxidative status. 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New approaches to selecting a scan-sampling method for chicken behavioral observations and their practical implications

The use of the scan-sampling method, especially when a large amount of data is collected, has become widespread in behavioral studies. However, there are no specific guidelines regarding the choice of the sampling interval in different conditions. Thus, establishing a standard approach for video analysis represents an important step forward within the scientific community. In the present work, we hypothesized that the length of the sampling interval could influence the results of chicken behavioral study, for which we evaluated the reliability, accuracy, and validity of three different sampling intervals (10, 15 and 30 min). The Bland-Altman test was proposed as an innovative approach to compare sampling intervals and support researcher choices. Moreover, these sampling intervals were applied to compare the behavior of 4 chicken genotypes kept under free-range conditions. The Bland-Altman plots suggested that sampling intervals greater than 10 min lead to biases in the estimation of rare behaviors, such as "Attacking". In contrast, the 30-min sampling interval was able to detect differences among genotypes in high-occurrence behaviors, such as those associated with locomotory activity. Thus, from a practical viewpoint, when a broad characterization of chicken genotypes is required, the 30-min scan-sampling interval might be suggested as a good compromise between resources and results

The Assessment of a Multifactorial Score for the Adaptability Evaluation of Six Poultry Genotypes to the Organic System

This study aimed to develop an adaptability score (AS) for chicken strains, which includes behavioral, plumage conditions, and body lesion indicators through a multifactorial approach. A total of 600 male chickens from 6 poultry genotypes—Ranger Classic (R1), Ranger Gold (R2), Rowan Ranger (R3), Hubbard Red JA (A), CY Gen 5 × JA87 (CY), and M22 × JA87 (M)—were reared under organic conditions, fed ad libitum, and individually weighed weekly to calculate the daily weight gain (DWG). The behavioral observations consisted of the explorative attitude (EA), recorded at 21 days, and the behavioral patterns (BPs) recorded the week before the slaughter. The AS was estab-lished by a principal component analysis, and the AS of these genotypes was compared. Moreover, the effect of DWG and genotype on the AS was evaluated by univariable and multivariable regres-sion models. Although the DWG and genotype were strictly dependent, genotype was the most important factor affecting the AS. In fact, its effect was significant both in univariable (p &lt; 0.001) and multivariable models (p &lt; 0.001). Conversely, the DWG was significant only in the univariable and lost significance when the effect of genotype was introduced in the model

The Assessment of a Multifactorial Score for the Adaptability Evaluation of Six Poultry Genotypes to the Organic System

This study aimed to develop an adaptability score (AS) for chicken strains, which includes behavioral, plumage conditions, and body lesion indicators through a multifactorial approach. A total of 600 male chickens from 6 poultry genotypes—Ranger Classic (R1), Ranger Gold (R2), Rowan Ranger (R3), Hubbard Red JA (A), CY Gen 5 × JA87 (CY), and M22 × JA87 (M)—were reared under organic conditions, fed ad libitum, and individually weighed weekly to calculate the daily weight gain (DWG). The behavioral observations consisted of the explorative attitude (EA), recorded at 21 days, and the behavioral patterns (BPs) recorded the week before the slaughter. The AS was established by a principal component analysis, and the AS of these genotypes was compared. Moreover, the effect of DWG and genotype on the AS was evaluated by univariable and multivariable regression models. Although the DWG and genotype were strictly dependent, genotype was the most important factor affecting the AS. In fact, its effect was significant both in univariable (p &lt; 0.001) and multivariable models (p &lt; 0.001). Conversely, the DWG was significant only in the univariable and lost significance when the effect of genotype was introduced in the model