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

UWB device for breast microwave imaging: phantom and clinical validations

Microwave imaging has received increasing attention in the last decades, motivated by its application in diagnostic imaging. Such effort has been encouraged by the fact that, at microwave frequencies, it is possible to distinguish between tissues with different dielectric properties. In such framework, a novel microwave device is presented here. The apparatus, consisting of two antennas operating in air, is completely safe and non-invasive since it does not emit any ionizing radiation and it can be used for breast lesion detection without requiring any breast crushing. We use Huygens Principle to provide a novel understanding into microwave imaging; specifically, the algorithm based on this principle provides images which represent homogeneity maps of the dielectric properties (dielectric constant and/or conductivity). The experimental results on phantoms having inclusions with different dielectric constants are presented here. In addition, the capability of the device to detect breast lesions has been verified through clinical examinations on 51 breasts. We introduce a metric to measure the non-homogeneous behaviour of the image, establishing a modality to detect the presence of inclusions inside phantoms and, similarly, the presence of a lesion inside a breast

Arsenic movement and fractionation in agricultural soils which received wastewater from an adjacent industrial site for 50 years

Arsenic (As) is an element with important environmental and human health implications due to its toxic properties. It is naturally occurring since it is contained in minerals, but it can also be enriched and distributed in the environment by anthropogenic activities. This paper reports on the historic As contamination of agricultural soils in one of the most important national relevance site for contamination in Italy, the so-called SIN Brescia-Caffaro, in the city of Brescia, northern Italy. These agricultural areas received As through the use of irrigation waters from wastewater coming from a factory of As-based pesticides (lead and calcium arsenates, sodium arsenite). Pesticide production started in 1920 and ended in the '70. Concentrations in the areas are generally beyond the legal threshold values for different soil uses and are up to >200 mg/kg. Arsenic contamination was studied to assess the long-time trend and the dynamics related to the vertical movement of As down to 1 m depth and its horizontal diffusion with surface irrigation in the entire field. Arsenic fractionation analysis (solid phase speciation by sequential extraction procedure) was also performed on samples collected from these areas and employed in greenhouse experiments with several plant species to evaluate the long-term contamination and the role of plant species in modifying As availability in soil. The results of this work can help in the evaluation of the conditions controlling the vertical transfer of As towards surface aquifers, the bioaccumulation likelihood in the agricultural food chain and the selection of sustainable remediation techniques such as phytoextraction

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

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%)

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

Mouse Panx1 Is Dispensable for Hearing Acquisition and Auditory Function

Panx1 forms plasma membrane channels in brain and several other organs, including the inner ear. Biophysical properties, activation mechanisms and modulators of Panx1 channels have been characterized in detail, however the impact of Panx1 on auditory function is unclear due to conflicts in published results. To address this issue, hearing performance and cochlear function of the Panx1−/− mouse strain, the first with a reported global ablation of Panx1, were scrutinized. Male and female homozygous (Panx1−/−), hemizygous (Panx1+/−) and their wild type (WT) siblings (Panx1+/+) were used for this study. Successful ablation of Panx1 was confirmed by RT-PCR and Western immunoblotting in the cochlea and brain of Panx1−/− mice. Furthermore, a previously validated Panx1-selective antibody revealed strong immunoreactivity in WT but not in Panx1−/− cochleae. Hearing sensitivity, outer hair cell-based “cochlear amplifier” and cochlear nerve function, analyzed by auditory brainstem response (ABR) and distortion product otoacoustic emission (DPOAE) recordings, were normal in Panx1+/− and Panx1−/− mice. In addition, we determined that global deletion of Panx1 impacts neither on connexin expression, nor on gap-junction coupling in the developing organ of Corti. Finally, spontaneous intercellular Ca2+ signal (ICS) activity in organotypic cochlear cultures, which is key to postnatal development of the organ of Corti and essential for hearing acquisition, was not affected by Panx1 ablation. Therefore, our results provide strong evidence that, in mice, Panx1 is dispensable for hearing acquisition and auditory function