Biomimetic Electrospun Self-Assembling Peptide Scaffolds for Neural Stem Cell Transplantation in Neural Tissue Engineering

Spinal cord regeneration using stem cell transplantation is a promising strategy for regenerative therapy. Stem cells transplanted onto scaffolds that can mimic natural extracellular matrix (ECM) have the potential to significantly improve outcomes. In this study, we strived to develop a cell carrier by culturing neural stem cells (NSCs) onto electrospun 2D and 3D constructs made up of specific crosslinked functionalized self-assembling peptides (SAPs) featuring enhanced biomimetic and biomechanical properties. Morphology, architecture, and secondary structures of electrospun scaffolds in the solid-state and electrospinning solution were studied step by step. Morphological studies showed the benefit of mixed peptides and surfactants as additives to form thinner, uniform, and defect-free fibers. It has been observed that β-sheet conformation as evidence of self-assembling has been predominant throughout the process except for the electrospinning solution. In vitro NSCs seeded on electrospun SAP scaffolds in 2D and 3D conditions displayed desirable proliferation, viability, and differentiation in comparison to the gold standard. In vivo biocompatibility assay confirmed the permissibility of implanted fibrous channels by foreign body reaction. The results of this study demonstrated that fibrous 2D/3D electrospun SAP scaffolds, when shaped as micro-channels, can be suitable to support NSC transplantation for regeneration following spinal cord injury

Data repurposing from digital home cage monitoring enlightens new perspectives on mouse motor behaviour and reduction principle

: In this longitudinal study we compare between and within-strain variation in the home-cage spatial preference of three widely used and commercially available mice strains-C57BL/6NCrl, BALB/cAnNCrl and CRL:CD1(ICR)-starting from the first hour post cage-change until the next cage-change, for three consecutive intervals, to further profile the circadian home-cage behavioural phenotypes. Cage-change can be a stressful moment in the life of laboratory mice, since animals are disturbed during the sleeping hours and must then rapidly re-adapt to a pristine environment, leading to disruptions in normal motor patterns. The novelty of this study resides in characterizing new strain-specific biological phenomena, such as activity along the cage walls and frontality, using the vast data reserves generated by previous experimental data, thus introducing the potential and exploring the applicability of data repurposing to enhance Reduction principle when running in vivo studies. Our results, entirely obtained without the use of new animals, demonstrate that also when referring to space preference within the cage, C57BL/6NCrl has a high variability in the behavioural phenotypes from pre-puberty until early adulthood compared to BALB/cAnNCrl, which is confirmed to be socially disaggregated, and CRL:CD1(ICR) which is conversely highly active and socially aggregated. Our data also suggest that a strain-oriented approach is needed when defining frequency of cage-change as well as maximum allowed animal density, which should be revised, ideally under the EU regulatory framework as well, according to the physiological peculiarities of the strains, and always avoiding the "one size fits all" approach

Oral D-Aspartate Treatment Improves Sperm Fertility in Both Young and Adult B6N Mice

D-Aspartate (D-Asp) treatment improved the fertility of young male C57BL/6N mice in vivo revealing a direct role on capacitation, acrosome reaction, and fertility in vitro in young males only. We investigated whether the positive effect of D-Asp on fertility could be extended to adult males and evaluated the efficacy of a 2- or 4-week-treatment in vivo. Therefore, 20 mM sodium D-Asp was supplied in drinking water to males of different ages so that they were 9 or 16 weeks old at the end of the experiments. After sperm freezing, the in vitro fertilization (IVF) rate, the birth rate, hormone levels (luteinizing hormone (LH), epitestosterone, and testosterone), the sperm quality (morphology, abnormalities, motility, and velocity), the capacitation rate, and the acrosome reaction were investigated. Oral D-Asp treatment improves the fertilizing capability in mice regardless of the age of the animals. Importantly, a short D-Asp treatment of 2 weeks in young males elevates sperm parameters to the levels of untreated adult animals. In vivo, D-Asp treatment highly improves sperm quality but not sperm concentration. Therefore, D-Asp plays a beneficial role in mouse male fertility and may be highly relevant for cryorepositories to improve mouse sperm biobanking

Comparison of in vivo and in vitro digestibility in donkeys

We compared in vivo and in vitro dry matter (DM) and neutral detergent fiber (NDF) digestibility in donkeys using feces as microbial inoculum. Four donkeys were used in a 4 × 4 Latin square design with a 2 × 2 factorial arrangement of treatments. The animals were fed two types of hay, with or without flaked barley. For the in vivo procedure, total feces were collected for 6 days from each donkey; digestibility was calculated as the difference between ingested and excreted DM and NDF. For the in vitro procedure, donkey feces were buffered and used as microbial inoculum in an Ankom DaisyII Incubator; digestibility was estimated after 60 h of incubation. In vivo results showed that the addition of barley to hays did not change the digestibility values. In vivo estimates were higher than in vitro ones. The equations used to predict in vivo estimates from in vitro data were not reliable (R2 = 0.47 and 0.21; P = 0.003 and 0.078 for NDF and DM digestibility, respectively). Further studies need to evaluate different sample size and digestion times

Towards large scale automated cage monitoring - Diurnal rhythm and impact of interventions on in-cage activity of C57BL/6J mice recorded 24/7 with a non-disrupting capacitive-based technique.

BACKGROUND AND AIMS: Automated recording of laboratory animal\u27s home cage behavior is receiving increasing attention since such non-intruding surveillance will aid in the unbiased understanding of animal cage behavior potentially improving animal experimental reproducibility. MATERIAL AND METHODS: Here we investigate activity of group held female C57BL/6J mice (mus musculus) housed in standard Individually Ventilated Cages across three test-sites: Consiglio Nazionale delle Ricerche (CNR, Rome, Italy), The Jackson Laboratory (JAX, Bar Harbor, USA) and Karolinska Insititutet (KI, Stockholm, Sweden). Additionally, comparison of female and male C57BL/6J mice was done at KI. Activity was recorded using a capacitive-based sensor placed non-intrusively on the cage rack under the home cage collecting activity data every 250 msec, 24/7. The data collection was analyzed using non-parametric analysis of variance for longitudinal data comparing sites, weekdays and sex. RESULTS: The system detected an increase in activity preceding and peaking around lights-on followed by a decrease to a rest pattern. At lights off, activity increased substantially displaying a distinct temporal variation across this period. We also documented impact on mouse activity that standard animal handling procedures have, e.g. cage-changes, and show that such procedures are stressors impacting in-cage activity. These key observations replicated across the three test-sites, however, it is also clear that, apparently minor local environmental differences generate significant behavioral variances between the sites and within sites across weeks. Comparison of gender revealed differences in activity in the response to cage-change lasting for days in male but not female mice; and apparently also impacting the response to other events such as lights-on in males. Females but not males showed a larger tendency for week-to-week variance in activity possibly reflecting estrous cycling. CONCLUSIONS: These data demonstrate that home cage monitoring is scalable and run in real time, providing complementary information for animal welfare measures, experimental design and phenotype characterization

Stocking density affects welfare indicators in horses reared for meat production

Horses kept for meat production are reared in intensive breeding farms. We employed a checklist adapted from the Animal Welfare Indicators (AWIN) assessment protocol. Our evaluation aims to assess whether welfare indicators are influenced by stocking densities (m2/horse) and feeding strategies applied. An analysis was carried out on the data obtained from 7 surveys conducted at a single horse farm designed for meat production. In each survey, the same 12 pens were assessed, but on each occasion, the horses in the pens had been changed as had the stocking densities. Briefly, 561 horses aged 16 ± 8 months (mean ± standard deviation) were evaluated. Two stocking density cut-off values (median and 75th percentile: 3.95 and 4.75 m2/horse, respectively) were applied to investigate the effect of stocking density on horse welfare. Data were analysed using Mann–Whitney U and Fisher’s exact tests (p < 0.05). When cut-off was set as the median percentile, lower stocking density was associated with improvements in body condition score (BCS), coat cleanliness and bedding quantity, less coughing, less resting in a standing position, and less feeding related to the greater space available at the feed bunk. When the 75th percentile cut-off was used, indicators that improved were coat cleanliness, bedding quantity and mane and tail condition, as well as less resting in standing position and less feeding related to the greater space available at the feed bunk. Accordingly, the use of two different stocking density cut-off values showed that the increase of space allowance affected specific welfare indicators. Further increment of space and/or changes in management regimes should be investigated to improve all the indicators. Moreover, results related to feeding indicated the need to intervene as starch intakes exceeded recommended safe levels, negatively affecting horse welfare

Time-budget of horses reared for meat production: Influence of stocking density on behavioural activities and subsequent welfare

Horses reared for meat production can be kept in intensive breeding farms where they are housed in group pens at high stocking densities. The present study aimed to evaluate whether the expressed behaviours correlated with stocking density, and to compare their time-budget with that of wild-living horses. An ethogram of 13 mutually exclusive behavioural activities was developed. Behavioural observations were performed over a 72 h period on group pens selected on the basis of stocking density and the homogeneity of breed, age, height at the withers, and time since arriving at the farm. Scan sampling (n = 96 scans/horse/day) was used on 22 horses. The mean frequency (%) standard deviation (SD) for each behavioural activity was calculated to obtain the time-budget. The associations between time-budget and stocking density were evaluated using a bivariate analysis. The relationships were analysed by Pearson’s correlation coecient (r). Our results show that locomotion, playing, and self-grooming positively correlated with a reduction in stocking density, indicating the potential to use these behaviours as positive welfare indicators for young horses kept in group pens. The data also revealed an unusual time-budget, where the main behavioural activity expressed was standing (30.56% 6.56%), followed by feeding (30.55% 3.59%), lying (27.33% 2.05%), and locomotion (4.07% 1.06%)

A Fibre- vs. cereal grain-based diet: Which is better for horse welfare? Effects on intestinal permeability, muscle characteristics and oxidative status in horses reared for meat production

Horses reared for meat production are fed high amounts of cereal grains in comparison with horses raised for other purposes. Such feeding practice may lead to risk of poor welfare consequences. The aim of this study was to investigate the effects of two feeding practices on selected metabolic parameters and production aspects. Nineteen Bardigiano horses, 14.3 ± 0.7 months of age, were randomly assigned to two groups—one fed with high amounts of cereal grains (HCG; n = 9; 43% hay plus 57% cereal grain-based pelleted feed) vs. one fed with high amounts of fibre (HFG; n = 10; 70% hay plus 30% pelleted fibrous feed)—for 129 days. At slaught on abattoir, biological and tissue samples were collected to evaluate the microbiological contamination of mesenteric lymph nodes and liver; selected meat quality traits (chemical composition and fatty acid profile of the Longissimus thoracis et lumborum muscle); and the oxidative status of the horse. A linear mixed model was used: dietary treatment and sex were fixed effects and their interaction analysed on production and metabolic parameters as dependent variables. Results showed an increased intestinal permeability in the horses fed HCG compared to HFG, according to the significant increased total mesophilic aerobic bacteria counts in mesenteric lymph nodes (p = 0.04) and liver samples (p = 0.05). Horses in HCG showed increased muscle pH (p = 0.02), lighter muscle colour (L) (p = 0.01), increased intramuscular fat concentrations (p = 0.03), increased muscle glutathione peroxidase and superoxide dismutase activities (p = 0.01 and p = 0.03, respectively). Moreover, horses in HCG had lower muscle water holding capacity at interaction with sex (p = 0.03, lower in female), lower muscle protein content (p = 0.01), lower concentration of muscle PUFAs (p = 0.05) and lower plasma catalase activities (p = 0.05). Our results showed that feeding a high cereal grains diet can have global effects on horse physiology, and thus represents a threat for their welfare

Geotechnical characterization of the upper Pleistocene-Holocene alluvial deposits of Roma (Italy) by means of multivariate geostatistics: Cross-validation results

We are presenting an attempt to evaluate the spatial variability of geotechnical parameters in the upper Pleistocene–Holocene alluvial deposits of Roma (Italy) by means of multivariate geostatistics. The upper Pleistocene–Holocene alluvial deposits of Roma are sensitive to high levels of geohazard. They occupy a sizable and significant part of the city, being the foundation for many monuments, historical neighborhoods, and archaeological areas, and the main host of the present and future subway lines. We have stored information from more than 2000 geotechnical boreholes crossing the alluvial deposits into a relational database. For the present study, only the boreholes with lithologic/textural interpretation and geotechnical information were selected. The set includes 283 boreholes and 719 samples, which have a set of geotechnical information comprising physical properties and mechanical parameters. Techniques of multivariate statistics and geostatistics were combined and compared to evaluate the estimation methods of the mechanical parameters, with special reference to the drained friction angle from direct shear test (φ′). Principal Component Analysis was applied to the dataset to highlight the relationships between the geotechnical parameters. Through cross-validation analysis, multiple linear regression, kriging, and cokriging were tested as estimators of φ′. Cross-validation demonstrates that the cokriging with granulometries as auxiliary variables is the most suitable method to estimate φ′. In addition to proving that cokriging is a good estimator of φ′, cross-validation demonstrates that input data are coherent and this allows us to use them for estimation of geotechnical parameters, although they come from different laboratories and different vintages. Nevertheless, to get the same good results of cross-validation in estimation, it is necessary for granulometries to be available at grid points. Since this information being not available at all grid points, it is expected that, in the future, textural information can be derived in an indirect way, i.e., from lithologic/textural spatial reconstructions.Published251-2682.3. TTC - Laboratori di chimica e fisica delle rocceJCR Journalope

Novel microwave apparatus for breast lesions detection: Preliminary clinical results

This paper presents preliminary results of an innovative microwave imaging apparatus for breast lesions detection. Specially, a Huygens Principle based method is employed to process the microwave signals and to build the respective microwave images. The apparatus has been first tested on phantoms. Next, its performance has been verified through clinical examinations on 22 healthy breasts and on 29 breast having lesions, using as gold standard the output of the radiologist study review obtained using conventional techniques. Specifically, we introduce a metric, which is the ratio between maximum and average of the image intensity (MAX/AVG). We found that MAX/AVG of microwave images can be used for classifying breasts containing lesions. In addition, using MAX/AVG as classification parameter, receiver operating characteristic curves have been empirically determined. Furthermore, for one randomly selected breast having lesion, we have demonstrated that the localisation of the inclusion acquired through microwave imaging is compatible with mammography images