Porcupettes Management at Wildlife Rescue Centers and Liberation into the Wild: Implications for Post-Liberation Success.

The crested porcupine is a frequent host species in wildlife rescue centers and no guidelines for its management, liberation and post-liberation monitoring are yet available. Here, captive-grown porcupines' behavior and survival in the wild after liberation were investigated and described for the first time. Management strategies adopted at the centers could affect porcupine adaptation to the natural environment. The detention of porcupettes in single cages and fed only human-supplied food may not ensure ethological welfare, nor a suitable behavioral development compatible with wildlife. The liberation of captive-grown porcupines should also be carefully planned to promote and increase the possibility of post-liberation success.

Exact Large-Scale Fluctuations of the Phase Field in the Sine-Gordon Model.

We present the first exact theory and analytical formulas for the large-scale phase fluctuations in the sine-Gordon model, valid in all regimes of the field theory, for arbitrary temperatures and interaction strengths. Our result is based on the ballistic fluctuation theory combined with generalized hydrodynamics, and can be seen as an exact "dressing" of the phenomenological soliton-gas picture first introduced by Sachdev and Young [Phys. Rev. Lett. 78, 2220 (1997)PRLTAO0031-900710.1103/PhysRevLett.78.2220], to the modes of generalized hydrodynamics. The resulting physics of phase fluctuations in the sine-Gordon model is qualitatively different, as the stable quasiparticles of integrability give coherent ballistic propagation instead of diffusive spreading. We provide extensive numerical checks of our analytical predictions within the classical regime of the field theory by using Monte Carlo methods. We discuss how our results are of ready applicability to experiments on tunnel-coupled quasicondensates.

Settlement of a stable wolf pack in a highly anthropic area of Pisan hills: Relationship with animal husbandry and hunting in a human-wolf coexistence perspective.

Anthropic areas play a pivot role for main wolf conservation challenges. Wolf presence in the higher Pisan hills has been well documented while wolf settlement in the lower Pisan hills is still uncertain. In this study, long-term information on wolf presence in a highly anthropic area of the lower Pisan hills was collected by using non-invasive monitoring techniques. Furthermore, both the relationship of this predator with human activity and the impact of hunting on wolf presence have been investigated. The results obtained indicate the presence of a stable and reproductive wolf pack composed by both Italian wolf and hybrids individuals in the municipalities of Crespina Lorenzana and Casciana Terme Lari. A high impact of wolf on livestock was recorded in this area since no prevention systems were adopted by farmers. Wolf appears not to have a negative impact on wild boar population. Similarly, wild boar drive hunting does not appear to affect the wolf pack presence in the area. Thereby wolf may play a key role as controller of wild population. Prevention strategies improvement becomes instrumental to promote wolf-human coexistence. Further investigation to monitor pack hybridization level and turnover and to assess the impact of packs on wild population and livestock in anthropic areas is desirable.

Evidence of scavenging behaviour in crested porcupine.

The vegetarian diet of many herbivorous mammals is supplemented with proteins of animal origin, especially in young individuals and in breeding females, to provide key proteins necessary for both growth and breeding. Among porcupine species, only the Cape porcupine (Hystrix africaeaustralis) has been observed to consume carrion flesh. From June to August 2019, a pigeon carcass was placed together with corn in 7 study settlements and near 2 monitored capture-traps, in order to assess the carrion flesh feeding habits of the crested porcupine (Hystrix cristata). Scavenging behaviour was recorded on four occasions. All the recorded individuals were adults and at least one was female. This demonstrates that the crested porcupine occasionally does eat flesh. Such evidence raises important questions concerning the relationship between feeding habits and the physiological needs of this herbivorous rodent.

Epidemiologic Survey on Toxoplasma gondii and Trichinella pseudospiralis Infection in Corvids from Central Italy.

Free-ranging corvids-678 magpies (Pica pica) and 120 hooded crows (Corvus cornix) from nine protected areas of the Pisa province (central Italy)-were examined for Toxoplasma gondii and Trichinella pseudospiralis. The intracardiac blood clots from 651 magpies and 120 hooded crows were serologically examined for T. gondii. The DNA extracted from the hearts of seropositive birds was then used to perform a nested PCR for the amplification of the T. gondii B1 gene and for genotyping for SAG genetic markers. Breast muscle samples from 678 magpies and 91 hooded crows were tested by an artificial digestion method for Trichinella. Data were statistically analyzed. Forty-five (5.8%-41 magpies and four hooded crows) out of the 771 examined animals scored seropositive for T. gondii, with titers ranging from 1:25 to 1:100. T. gondii DNA was detected in 15 of the 45 positive birds and T. gondii genotypes II and III were identified. No positivity for T. pseudospiralis was found. No significant differences between the two species of corvids and among the different areas of origin were observed for seropositivity to T. gondii. This is the first extensive study on both T. gondii and T. pseudospiralis in magpies and hooded crows, as well as the first detection of T. gondii SAG genotypes in magpies.

MASK-RL: Multiagent Video Object Segmentation Framework Through Reinforcement Learning.

Integrating human-provided location priors into video object segmentation has been shown to be an effective strategy to enhance performance, but their application at large scale is unfeasible. Gamification can help reduce the annotation burden, but it still requires user involvement. We propose a video object segmentation framework that leverages the combined advantages of user feedback for segmentation and gamification strategy by simulating multiple game players through a reinforcement learning (RL) model that reproduces human ability to pinpoint moving objects and using the simulated feedback to drive the decisions of a fully convolutional deep segmentation network. Experimental results on the DAVIS-17 benchmark show that: 1) including user-provided prior, even if not precise, yields high performance; 2) our RL agent replicates satisfactorily the same variability of humans in identifying spatiotemporal salient objects; and 3) employing artificially generated priors in an unsupervised video object segmentation model reaches state-of-the-art performance.

Diurnal motor activity and "sunbathing" behaviour in crested porcupine (Hystrix cristata L., 1758).

The crested porcupine is a mainly nocturnal mammal that shows both moonlight avoidance and some diurnal motor activity, the latter as an extension of its night-time foraging behaviour. Furthermore, a peculiar daytime behaviour, described as "sunbathing", was reported as episodic in H. africaeaustralis. Between 2016 and 2019 a camera-trapping monitoring was performed within 10 porcupine settlements in order to detect the diurnal motor activity and to verify and describe the sunbathing behaviour in crested porcupine. Out of 1,003 trap days, a total of 148 events of diurnal motor activity were recorded. The diurnal motor activity occurred throughout the year mainly between December and June from 15:00 to 16:00, with no statistical difference between cubs, youngsters and adults. The sunbathing behaviour was detected for a total of 36 episodes recorded. Sunbathing was performed mainly by cubs. The sunbathing behaviour occurred only between April and June during the hottest hours of the day (11:00 to 12:00). Diurnal motor activity and sunbathing behaviour of porcupine are discussed in relation to food availability and porcupine physiology.

The metabolic basis of cognitive insight in psychosis: A positron emission tomography study.

The purpose of this study was to investigate the relationship between cognitive insight and cerebral metabolism in patients suffering from psychosis. The Beck Cognitive Insight Scale (BCIS) was administered to 63 patients with psychosis undergoing Positron Emission Tomography investigation. The sample was divided into two groups considering the BCIS score. Data were analyzed using Statistical Parametric Mapping. RESULTS: patients with low insight, compared to those with high insight, showed decreased metabolism in the right fusiform gyrus, left precuneus, superior temporal gyrus and insula bilaterally, as well as increased metabolism in the left orbito-frontal gyrus (all p<0.005). Our results suggest that reduced posterior (occipito-temporo-insulo-parietal) and increased anterior (orbitofrontal) cerebral metabolism may sustain low cognitive insight in psychosis.

Cortico-Cerebellar Connectivity in Autism Spectrum Disorder: What Do We Know So Far?

Although the Autism Spectrum Disorder (ASD) is renowned to be a connectivity disorder and a condition characterized by cerebellar involvement, the connectivity between the cerebellum and other cortical brain regions is particularly underexamined. Indeed, converging evidence has recently suggested that the cerebellum could play a key role in the etiopathogenesis of ASD, since cerebellar anomalies have been consistently reported in ASD from the molecular to the behavioral level, and damage to the cerebellum early in development has been linked with signs of autistic features. In addition, current data have shown that the cerebellum is a key structure not only for sensory-motor control, but also for "higher functions," such as social cognition and emotion, through its extensive connections with cortical areas. The disruption of these circuits could be implicated in the wide range of autistic symptoms that the term "spectrum" connotes. In this review, we present and discuss the recent findings from imaging studies that investigated cortico-cerebellar connectivity in people with ASD. The literature is still too limited to allow for definitive conclusions; however, this brief review reveals substantial areas for future studies, underlining currently unmet research perspectives.

A hybrid chimeric system for versatile and ultra-sensitive RNase detection.

We developed a new versatile strategy that allows the detection of several classes of RNases (i.e., targeting ss- or ds-RNA, DNA/RNA hetero-hybrid or junctions) with higher sensitivity than existing assays. Our two-step approach consists of a DNA-RNA-DNA chimeric Hairpin Probe (cHP) conjugated to magnetic microparticles and containing a DNAzyme sequence in its terminal region, and molecular beacons for fluorescence signal generation. In the first step, the digestion of the RNA portion of the cHP sequences in presence of RNases leads to the release of multiple copies of the DNAzyme in solution. Then, after magnetic washing, each DNAzyme molecule elicits the catalytic cleavage of numerous molecular beacons, providing a strong amplification of the overall sensitivity of the assay. We successfully applied our approach to detect very low concentrations of RNase A, E. coli RNase I, and RNase H. Furthermore, we analyzed the effect of two antibiotics (penicillin and streptomycin) on RNase H activity, demonstrating the applicability of our strategy for the screening of inhibitors. Finally, we exploited our system to detect RNase activity directly in crude biological samples (i.e., blood and saliva) and in cell culture medium, highlighting its suitability as cheap and sensitive tool for the detection of RNase levels.

A fruit fly in the nanoworld: once again Drosophila contributes to environment and human health.

Drosophila was the most important model organism used in the fields of medicine and biology over the last century. Recently, Drosophila was successfully used in several studies in the field of nanotoxicology. However, only a part of its potential has been exploited in this field until now. In fact, apart from macroscopic observations of the effect due to the interaction between nanomaterials and living organism (i.e. lifespan, fertility, phenotypic aberrations, etc.), Drosophila has the potential to be a very useful tool to deeply analyze the molecular pathways involved in response to the interactions at nano-bio level. The aim of this editorial is to encourage the use of Drosophila by the different research groups working in the fields of nanotoxicology and nanomedicine, in order to define the effects induced by nanomaterials at molecular level for their subsequent exploitation in the field of nanomedicine.

A general mechanism for intracellular toxicity of metal-containing nanoparticles.

The assessment of the risks exerted by nanoparticles is a key challenge for academic, industrial, and regulatory communities worldwide. Experimental evidence points towards significant toxicity for a range of nanoparticles both in vitro and in vivo. Worldwide efforts aim at uncovering the underlying mechanisms for this toxicity. Here, we show that the intracellular ion release elicited by the acidic conditions of the lysosomal cellular compartment--where particles are abundantly internalized--is responsible for the cascading events associated with nanoparticles-induced intracellular toxicity. We call this mechanism a "lysosome-enhanced Trojan horse effect" since, in the case of nanoparticles, the protective cellular machinery designed to degrade foreign objects is actually responsible for their toxicity. To test our hypothesis, we compare the toxicity of similar gold particles whose main difference is in the internalization pathways. We show that particles known to pass directly through cell membranes become more toxic when modified so as to be mostly internalized by endocytosis. Furthermore, using experiments with chelating and lysosomotropic agents, we found that the toxicity mechanism for different metal containing NPs (such as metallic, metal oxide, and semiconductor NPs) is mainly associated with the release of the corresponding toxic ions. Finally, we show that particles unable to release toxic ions (such as stably coated NPs, or diamond and silica NPs) are not harmful to intracellular environments.

Lab-on-a-chip-based high-throughput screening of the genotoxicity of engineered nanomaterials.

The continuous increasing of engineered nanomaterials (ENMs) in our environment, their combinatorial diversity, and the associated genotoxic risks, highlight the urgent need to better define the possible toxicological effects of ENMs. In this context, we present a new high-throughput screening (HTS) platform based on the cytokinesis-block micronucleus (CBMN) assay, lab-on-chip cell sorting, and automated image analysis. This HTS platform has been successfully applied to the evaluation of the cytotoxic and genotoxic effects of silver nanoparticles (AgNPs) and silica nanoparticles (SiO2NPs). In particular, our results demonstrate the high cyto- and genotoxicity induced by AgNPs and the biocompatibility of SiO2NPs, in primary human lymphocytes. Moreover, our data reveal that the toxic effects are also dependent on size, surface coating, and surface charge. Most importantly, our HTS platform shows that AgNP-induced genotoxicity is lymphocyte sub-type dependent and is particularly pronounced in CD2+ and CD4+ cells.

InP/ZnS as a safer alternative to CdSe/ZnS core/shell quantum dots: in vitro and in vivo toxicity assessment.

We show that water soluble InP/ZnS core/shell QDs are a safer alternative to CdSe/ZnS QDs for biological applications, by comparing their toxicity in vitro (cell culture) and in vivo (animal model Drosophila). By choosing QDs with comparable physical and chemical properties, we find that cellular uptake and localization are practically identical for these two nanomaterials. Toxicity of CdSe/ZnS QDs appears to be related to the release of poisonous Cd(2+) ions and indeed we show that there is leaching of Cd(2+) ions from the particle core despite the two-layer ZnS shell. Since an almost identical amount of In(III) ions is observed to leach from the core of InP/ZnS QDs, their very low toxicity as revealed in this study hints at a much lower intrinsic toxicity of indium compared to cadmium.

Molecular response of Escherichia coli adhering onto nanoscale topography.

Bacterial adhesion onto abiotic surfaces is an important issue in biology and medicine since understanding the bases of such interaction represents a crucial aspect in the design of safe implant devices with intrinsic antibacterial characteristics. In this framework, we investigated the effects of nanostructured metal substrates on Escherichia coli adhesion and adaptation in order to understand the bio-molecular dynamics ruling the interactions at the interface. In particular, we show how highly controlled nanostructured gold substrates impact the bacterial behavior in terms of morphological changes and lead to modifications in the expression profile of several genes, which are crucially involved in the stress response and fimbrial synthesis. These results mainly demonstrate that E. coli cells are able to sense even slight changes in surface nanotopography and to actively respond by activating stress-related pathways. At the same time, our findings highlight the possibility of designing nanoengineered substrates able to trigger specific bio-molecular effects, thus opening the perspective of smartly tuning bacterial behavior by biomaterial design.

Concentration-dependent, size-independent toxicity of citrate capped AuNPs in Drosophila melanogaster.

The expected potential benefits promised by nanotechnology in various fields have led to a rapid increase of the presence of engineered nanomaterials in a high number of commercial goods. This is generating increasing questions about possible risks for human health and environment, due to the lack of an in-depth assessment of the physical/chemical factors responsible for their toxic effects. In this work, we evaluated the toxicity of monodisperse citrate-capped gold nanoparticles (AuNPs) of different sizes (5, 15, 40, and 80 nm) in the model organism Drosophila melanogaster, upon ingestion. To properly evaluate and distinguish the possible dose- and/or size-dependent toxicity of the AuNPs, we performed a thorough assessment of their biological effects, using two different dose-metrics. In the first approach, we kept constant the total surface area of the differently sized AuNPs (Total Exposed Surface area approach, TES), while, in the second approach, we used the same number concentration of the four different sizes of AuNPs (Total Number of Nanoparticles approach, TNN). We observed a significant AuNPs-induced toxicity in vivo, namely a strong reduction of Drosophila lifespan and fertility performance, presence of DNA fragmentation, as well as a significant modification in the expression levels of genes involved in stress responses, DNA damage recognition and apoptosis pathway. Interestingly, we found that, within the investigated experimental conditions, the toxic effects in the exposed organisms were directly related to the concentration of the AuNPs administered, irrespective of their size.

Mutagenic effects of gold nanoparticles induce aberrant phenotypes in Drosophila melanogaster.

The peculiar physical/chemical characteristics of engineered nanomaterials have led to a rapid increase of nanotechnology-based applications in many fields. However, before exploiting their huge and wide potential, it is necessary to assess their effects upon interaction with living systems. In this context, the screening of nanomaterials to evaluate their possible toxicity and understand the underlying mechanisms currently represents a crucial opportunity to prevent severe harmful effects in the next future. In this work we show the in vivo toxicity of gold nanoparticles (Au NPs) in Drosophila melanogaster, highlighting significant genotoxic effects and, thus, revealing an unsettling aspect of the long-term outcome of the exposure to this nanomaterial. After the treatment with Au NPs, we observed dramatic phenotypic modifications in the subsequent generations of Drosophila, demonstrating their capability to induce mutagenic effects that may be transmitted to the descendants. Noteworthy, we were able to obtain the first nanomaterial-mutated organism, named NM-mut. Although these results sound alarming, they underline the importance of systematic and reliable toxicology characterizations of nanomaterials and the necessity of significant efforts by the nanoscience community in designing and testing suitable nanoscale surface engineering/coating to develop biocompatible nanomaterials with no hazardous effects for human health and environment. FROM THE CLINICAL EDITOR: While the clinical application of nanomedicine is still in its infancy, the rapid evolution of this field will undoubtedly result in a growing number of clinical trials and eventually in human applications. The interactions of nanoparticles with living organisms determine their toxicity and long-term safety, which must be properly understood prior to large-scale applications are considered. The paper by Dr. Pompa's team is the first ever demonstration of mutagenesis resulting in clearly observable phenotypic alterations and the generation of nano-mutants as a result of exposure to citrate-surfaced gold nanoparticles in drosophila. These groundbreaking results are alarming, but represent a true milestone in nanomedicine and serve as a a reminder and warning about the critical importance of "safety first" in biomedical science.

SiO2 nanoparticles biocompatibility and their potential for gene delivery and silencing.

Despite the extensive use of silica nanoparticles (SiO(2)NPs) in many fields, the results about their potential toxicity are still controversial. In this work, we have performed a systematic in vitro study to assess the biological impact of SiO(2)NPs, by investigating 3 different sizes (25, 60 and 115 nm) and 2 surface charges (positive and negative) of the nanoparticles in 5 cell lines (3 in adherence and 2 in suspension). We analyzed the cellular uptake and distribution of the NPs along with their possible effects on cell viability, membrane integrity and generation of reactive oxygen species (ROS). Experimental results show that all the investigated SiO(2)NPs do not induce detectable cytotoxic effects (up to 2.5 nM concentration) in all cell lines, and that cellular uptake is mediated by an endocytic process strongly dependent on the particle size and independent of its original surface charge, due to protein corona effects. Once having assessed the biocompatibility of SiO(2)NPs, we have evaluated their potential in gene delivery, showing their ability to silence specific protein expression. The results of this work indicate that monodisperse and stable SiO(2)NPs are not toxic, revealing their promising potential in various biomedical applications.

Impact of nanoscale topography on genomics and proteomics of adherent bacteria.

Bacterial adhesion onto inorganic/nanoengineered surfaces is a key issue in biotechnology and medicine, because it is one of the first necessary steps to determine a general pathogenic event. Understanding the molecular mechanisms of bacteria-surface interaction represents a milestone for planning a new generation of devices with unanimously certified antibacterial characteristics. Here, we show how highly controlled nanostructured substrates impact the bacterial behavior in terms of morphological, genomic, and proteomic response. We observed by atomic force microscopy (AFM) and scanning electron microscopy (SEM) that type-1 fimbriae typically disappear in Escherichia coli adherent onto nanostructured substrates, as opposed to bacteria onto reference glass or flat gold surfaces. A genetic variation of the fimbrial operon regulation was consistently identified by real time qPCR in bacteria interacting with the nanorough substrates. To gain a deeper insight into the molecular basis of the interaction mechanisms, we explored the entire proteomic profile of E. coli by 2D-DIGE, finding significant changes in the bacteria adherent onto the nanorough substrates, such as regulations of proteins involved in stress processes and defense mechanisms. We thus demonstrated that a pure physical stimulus, that is, a nanoscale variation of surface topography, may play per se a significant role in determining the morphological, genetic, and proteomic profile of bacteria. These data suggest that in depth investigations of the molecular processes of microorganisms adhering to surfaces are of great importance for the design of innovative biomaterials with active biological functionalities.

Correlation between body mass index and others risk factors for cardiovascular disease in women compared with men.

OBJECTIVE: In this study we examine the association between body mass index and a wide range of metabolic coronary heart disease risk factors in a female population of South Italy. DESIGN: Observational study named "VIP Project" divided in three phases: collection of data, follow-up of the population and new controls within five and ten years. Data presented are about the transversal phase of the study. SUBJECTS: 1200, 600 males and 600 females, age ranging from 25 to 74 years, were enrolled at random from the electoral lists of the towns of Mercato San Severino and Baronissi, near Salerno, in Southern Italy. MEASUREMENTS: Weight (electronic scale), height (ruler attached to the wall), blood pressure (sphygmomanometer), fasting venous blood to determine: total cholesterol, HDL and LDL cholesterol, triglycerides, uric acid, blood glucose, haemocrome, insulinaemia, fibrinogen, C3 and creatinine. Smoking habit and the practice of sports were determined by an interview. RESULTS: The distribution of BMI in the population, increases with reference to the age. Between BMI and the classical risk factors, there is a significant correlation with triglycerides, glycaemia, total cholesterol, diastolic blood pressure, systolic blood pressure, insulin, C3. In the decades from 35-44 years to 45-54 years, it can be observed a notable increasing of the following risk factors in percentage for female population: hypertriglyceridemia from 3.9% to 33.9%, diabetes from 4.4% to 10.2%, hypertension from 9.1% to 25.8%, obesity from 22.5% to 42.5% and hypercholesterolaemia from 3.6% to 25%. Only the number of smokers is decreasing from 38.3% to 19.2%. CONCLUSION: The association between body mass index and increasing coronary heart disease risk in women is partly explained by a rise in blood pressure, lipid profile and blood glucose across the range of body mass index. Most approaches to weight loss recommend a target or optimal weight based on body mass index alone. Our data show that this assumption is unwarranted and that body mass index should not be used as the sole basis for intervention in individuals. Successful weight loss should be defined in terms of a reduction in metabolic risk, which can often be achieved by relatively modest weight loss.