How COVID-19 Pandemic Changed Children and Adolescents Use of the Emergency Department: the Experience of a Secondary Care Pediatric Unit in Central Italy.

Italy was the first European country hit by SARS-CoV-2 infection, particularly northern regions. After the beginning of national lockdown (March 9th, 2020), we observed a significant decrease in pediatric emergency department consultations (daily pediatric visits; pre-lockdown, 16 (11-22); lockdown, 3 (1-3); phase 2, 3 (3-5), p < 0.0001). On the other hand, the percentage of children discharged right after pediatric visit significantly decreased from 80% in January to 50% in April. After March 9th, we registered a change in the diagnoses of emergency department visits, with an increase in the percentage of non-infectious acute conditions and a decrease in infectious diseases, with two cases of a noteworthy delayed access to hospital care. We performed a retrospective analysis of consultations requested to our pediatric unit for children and adolescents referred to the general Emergency Department of San Luca Hospital of Lucca (Tuscany, Central Italy) from January 1st to May 31st, 2020. We split data in two different time periods according to consultations performed before (January 1st-March 8th) and after the beginning of lockdown (March 9th-May 31st). Analyzing the number of children hospitalized from January to May 2020 in comparison with the same period in 2019, a decreased hospitalization became evident after March (March - 74.6%, April - 71.6%, May - 58.6%). Nasopharyngeal swabs done in 115 children showed only one case of COVID-19. Even if COVID-19 outbreak more seriously affected Northern Italy, utilization of pediatric emergency services significantly changed also in Central Italy with consequent reduced demand and increased appropriateness.

Transcriptome characterization and expression profiling in chestnut cultivars resistant or susceptible to the gall wasp Dryocosmus kuriphilus.

The oriental gall wasp Dryocosmus kuriphilus represents a limiting pest for the European Chestnut (Castanea sativa, Fagaceae) as it creates severe yield losses. The European Chestnut is a deciduous tree, having major social, economic and environmental importance in Southern Europe, covering an area of 2.53 million hectares, including 75,000 ha devoted to fruit production. Cultivars show different susceptibility and very few are resistant to gall wasp. To deeply investigate the plant response and understand which factors can lead the plant to develop or not the gall, the study of transcriptome is basic (fundamental). To date, little transcriptomic information are available for C. sativa species. Hence, we present a de novo assembly of the chestnut transcriptome of the resistant Euro-Japanese hybrid 'Bouche de Bétizac' (BB) and the susceptible cultivar 'Madonna' (M), collecting RNA from buds at different stages of budburst. The two transcriptomes were assembled into 34,081 (BB) and 30,605 (M) unigenes, respectively. The former was used as a reference sequence for further characterization analyses, highlighting the presence of 1444 putative resistance gene analogs (RGAs) and about 1135 unigenes, as putative MiRNA targets. A global quantitative transcriptome profiling comparing the resistant and the susceptible cultivars, in the presence or not of the gall wasp, revealed some GO enrichments as "response to stimulus" (GO:0050896), and "developmental processes" (e.g., post-embryonic development, GO:0009791). Many up-regulated genes appeared to be transcription factors (e.g., RAV1, AP2/ERF, WRKY33) or protein regulators (e.g., RAPTOR1B) and storage proteins (e.g., LEA D29) involved in "post-embryonic development". Our analysis was able to provide a large amount of information, including 7k simple sequence repeat (SSR) and 335k single-nucleotide polymorphism (SNP)/INDEL markers, and generated the first reference unigene catalog for the European Chestnut. The transcriptome data for C. sativa will contribute to understand the genetic basis of the resistance to gall wasp and will provide useful information for next molecular genetic studies of this species and its relatives.

Combining porphyrins and pH indicators for analyte detection.

High sensitivity and cross-selectivity are mandatory properties for sensor arrays. Although metalloporphyrins and pH indicators are among the most common and appropriate choices for the preparation of optical sensor arrays, the sensitivity spectrum of these dyes is limited to those analytes able to induce an optical response. To extend the receptive field of optical sensors, we explore the design of composite materials, where the molecular interaction among the subunits enriches their sensing working mechanisms. We demonstrate that blends of single metalloporphyrins and pH indicators, tested with a transduction apparatus based on ubiquitous and easily available hardware, can be endowed with sensing properties wider than those of single constituents, enabling the recognition of a broad range of volatiles.

Volatile emissions from compressed tissue.

Since almost every fifth patient treated in hospital care develops pressure ulcers, early identification of risk is important. A non-invasive method for the elucidation of endogenous biomarkers related to pressure ulcers could be an excellent tool for this purpose. We therefore found it of interest to determine if there is a difference in the emissions of volatiles from compressed and uncompressed tissue. The ultimate goal is to find a non-invasive method to obtain an early warning for the risk of developing pressure ulcers for bed-ridden persons. Chemical analysis of the emissions, collected in compresses, was made with gas-chromatography-mass spectrometry and with a chemical sensor array, the so called electronic nose. It was found that the emissions from healthy and hospitalized persons differed significantly irrespective of the site. Within each group there was a clear difference between the compressed and uncompressed site. Peaks that could be certainly deemed as markers of the compression were, however, not identified. Nonetheless, different compounds connected to the application of local mechanical pressure were found. The results obtained with GC-MS reveal the complexity of VOC composition, thus an array of non-selective chemical sensors seems to be a suitable choice for the analysis of skin emission from compressed tissues; it may represent a practical instrument for bed side diagnostics. Results show that the adopted electronic noses are likely sensitive to the total amount of the emission rather than to its composition. The development of a gas sensor-based device requires then the design of sensor receptors adequate to detect the VOCs bouquet typical of pressure. This preliminary experiment evidences the necessity of studies where each given person is followed for a long time in a ward in order to detect the insurgence of specific VOCs pattern changes signalling the occurrence of ulcers.

mTrop1/Epcam knockout mice develop congenital tufting enteropathy through dysregulation of intestinal E-cadherin/ß-catenin.

Congenital tufting enteropathy (CTE) is a life-threatening hereditary disease that is characterized by enteric mucosa tufting degeneration and early onset, severe diarrhea. Loss-of-function mutations of the human EPCAM gene (TROP1, TACSTD1) have been indicated as the cause of CTE. However, loss of mTrop1/Epcam in mice appeared to lead to death in utero, due to placental malformation. This and indications of residual Trop-1/EpCAM expression in cases of CTE cast doubt on the role of mTrop1/Epcam in this disease. The aim of this study was to determine the role of TROP1/EPCAM in CTE and to generate an animal model of this disease for molecular investigation and therapy development. Using a rigorous gene-trapping approach, we obtained mTrop1/Epcam -null (knockout) mice. These were born alive, but failed to thrive, and died soon after birth because of hemorrhagic diarrhea. The intestine from the mTrop1/Epcam knockout mice showed intestinal tufts, villous atrophy and colon crypt hyperplasia, as in human CTE. No structural defects were detected in other organs. These results are consistent with TROP1/EPCAM loss being the cause of CTE, thus providing a viable animal model for this disease, and a benchmark for its pathogenetic course. In the affected enteric mucosa, E-cadherin and ß-catenin were shown to be dysregulated, leading to disorganized transition from crypts to villi, with progressive loss of membrane localization and increasing intracellular accumulation, thus unraveling an essential role for Trop-1/EpCAM in the maintenance of intestinal architecture and functionality.Supporting information is available for this article.

Detection of a hypersensitive reaction in the chestnut hybrid 'Bouche de Bétizac' infested by Dryocosmus kuriphilus Yasumatsu.

The purpose of the study was the identification of the mechanisms of resistance to Dryocosmus kuriphilus Yasumatsu in the hybrid-resistant cultivar 'Bouche de Bétizac' (Castanea sativa × Castanea crenata). Larvae and eggs of the insect are found in the buds of this cultivar at the end of winter, but there is no gall development after budburst. The hypothesis of the presence of a hypersensitive reaction (HR) in the buds was tested using diaminobenzidine (DAB) to detect H(2)O(2) and by Real Time PCR (RT-PCR) to evaluate the expression of a germin-like protein gene. HR in plants is elicited by the production of reactive oxygen compounds, such as H(2)O(2), and results in the programmed cell death. The DAB test was applied to buds of 'Bouche de Bétizac' and of the susceptible cultivar 'Madonna' (C. sativa) at different stages of budburst. The DAB staining produced brown areas in the swelling buds of 'Bouche de Bétizac', indicating the presence of H(2)O(2). On the contrary, all uninfested buds, as well as the infested buds of 'Madonna', appeared whitish. Papers report that germin and germin-like proteins (GLP) with oxalate oxidase activity are discrete markers of stress-responsive gene products. A strong expression of the chestnut GLP gene was detected by RT-PCR at bud swelling in infested 'Bouche de Bétizac' buds but not in 'Madonna' ones. The results support the hypothesis of the occurrence of an HR in 'Bouche de Bétizac' as response to the cynipid infestation, resulting in cell and larvae death.

Data processing for image-based chemical sensors: unsupervised region of interest selection and background noise compensation.

Natural olfaction suggests that numerous replicas of small sensors can achieve large sensitivity. This concept of sensor redundancy can be exploited by use of optical chemical sensors whose use of image sensors enables the simultaneous measurement of several spatially distributed indicators. Digital image sensors split the framed scene into hundreds of thousands of pixels each corresponding to a portion of the sensing layer. The signal from each pixel can be regarded as an independent sensor, which leads to a highly redundant sensor array. Such redundancy can eventually be exploited to increase the signal-to-noise ratio. In this paper we report an algorithm for reduction of the noise of pixel signals. For this purpose, the algorithm processes the output of groups of pixels whose signals share the same time behavior, as is the case for signals related to the same indicator. To define these groups of pixels, unsupervised clustering, based on classification of the indicator colors, is proposed here. This approach to signal processing is tested in experiments on the chemical sensitivity of replicas of eight indicators spotted on to a plastic substrate. Results show that the groups of pixels can be defined independently of the geometrical arrangement of the sensing spots, and substantial improvement of the signal-to-noise ratio is obtained, enabling the detection of volatile compounds at any location on the distributed sensing layer.

An investigation on the role of spike latency in an artificial olfactory system.

Experimental studies have shown that the reactions to external stimuli may appear only few hundreds of milliseconds after the physical interaction of the stimulus with the proper receptor. This behavior suggests that neurons transmit the largest meaningful part of their signal in the first spikes, and than that the spike latency is a good descriptor of the information content in biological neural networks. In this paper this property has been investigated in an artificial sensorial system where a single layer of spiking neurons is trained with the data generated by an artificial olfactory platform based on a large array of chemical sensors. The capability to discriminate between distinct chemicals and mixtures of them was studied with spiking neural networks endowed with and without lateral inhibitions and considering as output feature of the network both the spikes latency and the average firing rate. Results show that the average firing rate of the output spikes sequences shows the best separation among the experienced vapors, however the latency code is able in a shorter time to correctly discriminate all the tested volatile compounds. This behavior is qualitatively similar to those recently found in natural olfaction, and noteworthy it provides practical suggestions to tail the measurement conditions of artificial olfactory systems defining for each specific case a proper measurement time.

Fluorescence based sensor arrays.

Fluorescence-based cross reactive sensor arrays have experienced significant development in the last decade because of the advantages that they can offer with respect to other transduction mechanisms, in terms of the usual performance parameters such as sensitivity, selectivity and so on. From this point of view, a great impulse to this development has been due to the realization of novel transduction platforms, which has also taken advantage of the development of consumer electronics such as digital scanners, cameras, and screens, allowing the realization of low cost sensing layers suitable for many practical applications. This possibility, combined with continuous optimization of sensing material properties, the possible preparation of arrays with a high number of individual sensing elements and pattern recognition data analysis, has led to novel opportunities for the creation of luminescence based sensor arrays with improved capabilities. Herein we report on the development of these devices witnessed in the last decade, dividing the developed devices according to their exploitation in gaseous or in solution phase.

Poly(ester urethane) guides for peripheral nerve regeneration.

A biocompatible and elastomeric PU was synthesized from low-molecular-weight PCL as macrodiol, CMD as chain extender and HDI as chain linker for applications in the field of peripheral nerve repair. PU cast films supported in vitro attachment and proliferation of NOBEC. The in vitro adhesion and proliferation of S5Y5 neuroblastoma cells on the inner surface of uncoated, gelatin- and PL-coated PU guides were compared. Due to their superior in vitro performance, PL-coated PU guides were tested in vivo for the repair of 1.8 cm-long defects in rat sciatic nerves. The progressive regeneration was confirmed by EMG and histological analysis showing the presence of regenerating fibers in the distal stumps.

Polymers with embedded chemical indicators as an artificial olfactory mucosa.

Physiological investigations suggest that the olfactory mucosa probably plays an ancillary role in the recognition of odours introducing a sort of chromatographic separation that, together with the zonal distribution of olfactory receptors, gives place to selective spatio-temporal response patterns. It has been recently suggested that this behaviour may be simulated by chemical sensors embedded in continuous polymer layers. In this paper, in analogy to the biology of olfaction, a simple and compact platform able to separate and detect gases and vapours on the basis of their diffusion properties is proposed. In such a system, broadly selective colour indicators, such as metalloporphyrins, are embedded in continuous layers of polymers with different sorption properties. The exposure to various alcohols and amines shows that the porphyrins are mainly responsible for the recognition of the molecular family, while the occurring spatio-temporal signal patterns make possible the identification of the individual chemical species.

Chemical sensitivity of self-assembled porphyrin nano-aggregates.

Nanostructured molecular assemblies may provide additional sensing properties not found in other arrangements of the same basic constituents. Among three-dimensional structures, nanotubes are particularly appealing for applications as chemical sensors, because of the potential inclusion of different guests inside the cavity or the induced modification of the skeletal interaction after analyte binding. Porphyrins are a class of compounds characterized by brilliant sensing properties, appearing also in non-ordered solid-state aggregates. In recent years, it was reported that aggregation of oppositely charged porphyrins led to the formation of self-assembled nanotubes and in this paper their sensing properties, both in solution and in the solid state, have been investigated. The interactions of porphyrin nanotubes with guest molecules have been monitored by following the changes in their UV-vis spectra. The results obtained have been exploited to build up a sensing platform based on a computer screen as a light source and a digital camera as detector. Porphyrin nanostructures exhibited an enhanced sensitivity to different compounds with respect to those shown by single porphyrin subunits. The reason for the increased sensitivity may be likely found in an additional sensing mechanism related to the modulation of the strength of the forces that keep the supramolecular ensemble together.

Melt-extruded guides for peripheral nerve regeneration. Part I: poly(epsilon-caprolactone).

Melt-extruded guides for peripheral nerve repair based on poly(epsilon-caprolactone) (PCL) were realised and their physico-chemical properties were evaluated. Preliminarily, PCL cast films were found to support the attachment and proliferation of Neonatal Olfactory Bulb Ensheating Cells (NOBEC). S5Y5 neuroblastoma cells were cultured inside PCL guides in their uncoated form or coated with a non-specific adhesion protein (gelatin) and a specific peptide for nerve regeneration (poly(L-lysine)). Coating increased cell density (gelatin) and/or the cell density rate on substrates (poly(L-lysine); gelatin) as compared to uncoated guides. Various in vivo tests were carried out for the repair of small (0.5 cm), medium (1.5 cm) and long (4.5 cm) size defects in the peripheral nerves of Wistar rats. For the small nerve defects, uncoated and coated PCL guides were tested. Results from in vivo tests were subjected to histological examination after 45 days, 6 and 8 months postoperative for small, medium and large defects, respectively. Regeneration was found for small and medium size defects. For 0.5 cm defects, the coating did not affect regeneration significantly. Grip-tests also evidenced functional recovery for the 1.5 cm-long defects treated with PCL guides, after 6 months from implantation. On the other hand, mechanical stiffness of PCL conduits impaired the repair of 4.5 cm-long defects in 8-month period: the lack of flexibility of the guide to rat movements caused its detachment from the implant site. The research showed that PCL guides can be used for the successful repair of small and medium size nerve defects, with possible improvements by suitable bio-mimetic coatings.

Clinical analysis of human urine by means of potentiometric Electronic tongue.

The Electronic tongue (ET) composed of different kind of potentiometric chemical sensors has been applied for the detection of urinary system dysfunctions and creatinine levels. The creatinine contents evaluated by ET were compared with those obtained by automated Jaffe's method and GC-MS, obtaining a satisfying agreement for both methods. Partial least square regression discriminate analysis (PLS-DA) and feed forward back-propagation neural network (FFBP NN) classified 51 urine specimens from healthy volunteers in four classes, according to the creatinine content, showing that both techniques can satisfactorily differentiate urines according to this parameter. The best accuracy result of 92.2% correct classification of unknown samples was achieved with FFBP NN. Moreover, the possibility of ET system to distinguish between urine samples of healthy patients, and those with malignant and non-malignant tumor diagnosis of bladder has been shown.

[Venous thromboembolism risk in medical inpatients]. / Rischio di tromboembolismo venoso nel paziente internistico.

Venous thromboembolism is one of the main causes of morbidity and mortality in many diseases, either in medical or in surgical wards. The aim of this study is to determine its frequency in a medical ward and to find all the patients who need a prophylaxys. We examined all the inpatients admitted in our Department during a period of three years, selecting all subjects affected by venous thromboembolism. Patients affected by metastatic cancer, namely pancreatic one, or congestive heart failure showed higher risk to develope venous thromboembolism. A prompt heparinic prophylaxys could reduce thromboembolic events. Male gender could be a further risk factor.

PAM-microfabricated polyurethane scaffolds: in vivo and in vitro preliminary studies.

Polymeric scaffolds were realised with linear degradable PU in the form of square, hexagonal and octagonal grids. They were characterised in terms of their mechanical properties. Analysis shows that the mechanical properties of the scaffolds depend on their geometries which are easily modulated using PAM. In vitro biological assays showed that PU promotes the adhesion and proliferation of fibroblast cells and that cell activities are better on PU scaffolds than on PU films. In vivo implantation of PU and PLGA scaffolds and PU films demonstrated that the scaffolds are completely resorbed after three months with a slight inflammatory response, while the PU film was still present after six months with an intense granulomatous reaction.

Suspension of bone marrow-derived undifferentiated mesenchymal stromal cells for repair of superficial digital flexor tendon in race horses.

It has been proven that mesenchymal stromal cells (MSCs) can differentiate into tenocytes. Attempts to repair tendon lesions have been performed, mainly using scaffold carriers in experimental settings. In this article, we describe the clinical use of undifferentiated MSCs in racehorses. Significant clinical recovery was achieved in 9 of 11 horses evaluated using ultrasound analysis and their ability to return to racing. Our results show that the suspension of a small number of undifferentiated MSCs may be sufficient to repair damaged tendons without the use of scaffold support. Ultrasound scanning showed that fibers were correctly oriented. By using undifferentiated cells, no ectopic bone deposition occurred. A sufficient number of cells was recovered for therapeutic purposes in all but 1 case. We suggest that the use of autologous MSCs is a safe therapeutic method for treating incompletely (i.e., not full-thickness) damaged tendons.

Seroprevalence of varicella zoster and rubella antibodies among rural populations of the Chaco region, south-eastern Bolivia.

OBJECTIVE: To determine the seroprevalence of antibodies against varicella zoster virus (VZV) and rubella virus among the population of two rural areas, Camiri and Villa Montes, Chaco region, south-eastern Bolivia. METHODS: In a cross-sectional study, serum samples from randomly selected subjects were screened for VZV- and rubella antibodies using enzyme-linked immunosorbent assay (ELISA). RESULTS: The prevalence of VZV antibodies was 80% (391 of 489). No significant differences were observed between genders and study areas. The prevalence increased with age with a seropositivity rate of 21.2% in the 1-4-year-old children, 56.9% in the 5-9 age group and 83.7% in the 10-14 age group, and reached 98.2% in over 45-year olds. The prevalence of rubella virus antibodies was 76.9% (377 of 490), without significant differences between genders and study areas. Similar to the trend observed for VZV antibodies, the seroprevalence increased with age with a seropositivity rate of 18.1% in the 1-4-year-old children, 53.9% in the 5-9 age group and 78.4% in the 10-14 age group, and reached 94.7% in over 45-year-old subjects. Among the 95 women of childbearing age (15-44 years) the susceptibility rate was 11.6%. CONCLUSIONS: A common seroimmunological profile was evidenced for the two infections. The age-specific profile of VZV seropositivity differs from that reported for other tropical countries where higher median age of seroconversion and lower seroprevalence among adults are observed. Data concerning the prevalence of immunity to rubella may be useful to evaluate the impact of the recently introduced rubella immunization programme.