On the morphology of Anisakis pegreffii: a comparative analysis of three microscopic techniques used to build a new parasite atlas

BACKGROUND: Human anisakidosis is a parasitic anthropozoonosis caused by larval nematodes of the family Anisakidae. Here, we report a detailed description of the morphology of Anisakis pegreffii third-stage larva performed using a conventional light and confocal microscopy, and scanning electron microscopy (SEM) that provide a basis for both phenotypic studies and genetic mutations. METHODS: The collected larvae from fish were morphologically identified as Anisakis larvae Type I, and they were characterized by PCR-RFLP to identify the Anisakis pegreffii specie. Using NC5/NC2 primers, ribosomal genomic regions ITS1, 5.8 SrRNA and ITS2 of DNA were amplified and PCR products were sequenced. Fifteen larvae belonging to Anisakis pegreffii were fixed, sectioned, and examined with a light and confocal microscope and by SEM. RESULTS: In our studies, have been acquired detailed ultrastructural images, which have been integrated with those derived from the dissection of the parasite, obtained with light and confocal microscopy. The structural and ultrastructural images concerning the third stage larvae of Anisakis pegreffii have been studied, analyzed and compared among them. The derived overall view has allowed detecting new interesting details of a well-known parasite and has been schematically showed. CONCLUSIONS: The aim of this study is to furnish an updated atlas of Anisakis pegreffii. Confocal microscopy, as well as the light and electron microscopy have played a pivotal role in the accumulation of new scientific data regarding the anatomical structures of this nematode. This work is the result of one year of engagement by the Authors and the outcome is a comprehensive atlas on Anisakis pegreffii microscopy

Human-Friendly Robotics 2020

Humans are very effective at interpreting subtle properties of the partner\u2019s movement and use this skill to promote smooth interactions. Therefore, robotic platforms that support human partners in daily activities should acquire similar abilities. In this work we focused on the features of human motor actions that communicate insights on the weight of an object and the carefulness required in its manipulation. Our final goal is to enable a robot to autonomously infer the degree of care required in object handling and to discriminate whether the item is light or heavy, just by observing a human manipulation. This preliminary study represents a promising step towards the implementation of those abilities on a robot observing the scene with its camera. Indeed, we succeeded in demonstrating that it is possible to reliably deduct if the human operator is careful when handling an object, through machine learning algorithms relying on the stream of visual acquisition from either a robot camera or from a motion capture system. On the other hand, we observed that the same approach is inadequate to discriminate between light and heavy objects

In vitro degradation and bioactivity of composite poly-l-lactic (PLLA)/bioactive glass (BG) scaffolds: comparison of 45S5 and 1393BG compositions

The objective of this study was to compare the effect of two bioglass (BG) compositions 45S5 and 1393 in poly-l-lactic composite scaffolds in terms of morphology, mechanical properties, biodegradation, water uptake and bioactivity. The scaffolds were produced via thermally induced phase separation starting from a ternary polymer solution (polymer/solvent/non-solvent). Furthermore, different BG to polymer ratios have been selected (1, 2.5, 5% wt/wt) to evaluate the effect of the amount of filler on the composite structure. Results show that the addition of 1393BG does not affect the scaffold morphology, whereas the 45S5BG at the highest amount tends to appreciably modify the scaffold architecture interacting with the phase separation process. Bioactivity tests confirmed the formation of a hydroxycarbonateapatite-layer in both types of BGs (detected via scanning electron microscopy, X-ray diffractometry and Fourier Transform Infrared Spectroscopy). Overall, the results showed that 1393BG composition affects the experimental preparation protocol to a minimal extent thus allowing a better control of the scaffold\ue2\u80\u99s morphology compared to 45S5BG

Kefiran-based scaffolds for biomedical applications

Kefiran is an exopolysaccharide produced by microorganisms present in kefir grains, with several health promoting properties. A optimized protocol was developed for the separation of kefiran from kefir grains, allowing to reach a yield 4\uf75 % without using toxic or expensive chemicals. The capability of kefiran to produce scaffold via Thermally Induced Phase Separation (TIPS) technique was investigated and porous scaffolds structure was obtained. Separated kefiran and scaffolds were analyzed via DSC and different thermal properties between purified kefiran and scaffold were revealed. XRD analysis revealed different structure between kefiran and scaffolds. The porous scaffold structure can be modulated by modifiying the thermal path of the solution during the phase separation. The citotoxicity of kefiran was evaluated with cell viability test and a significant faster cell proliferation in the treated colture medium with kefiran was evidenced

3D cultures of rat astrocytes and brain capillary endothelial cells on Poly-L-lactic acid scaffolds

Tissue engineering is an emerging multidisciplinary field that aims at reproducing in vitro and/or in vivo tissues with morphological and functional features similar to the biological tissue of the human body. In this communication we report setting of three-dimensional structures able to mimic the extracellular matrix of the nervous system: we prepared Poly-L-Lactic Acid (PLLA) porous scaffolds via thermally induced phase separation (TIPS), and investigated the parameters that influence porosity, average pore size and degree of interconnection, i.e. polymer concentration, temperature and time of process. Astrocytes and brain capillary endothelial cells (BCECs) were cultured on these three-dimensional structures and tested for their ability to grow and survive on PLLA scaffolds. We analyzed in parallel the cell growth in 2D and 3D culture systems and observed the differences in cell morphology by fluorescence analysis: three-dimensional scaffolds have the ability to guide cell growth, provide support, encourage cell adhesion and proliferation. Astrocytes and BCECs adapted well to these porous matrices, not only remaining on the surface, but also penetrating inside the scaffolds. This 3D cell culture system could be further enriched to host two or three different brain cell types, in order to set an in vitro model of blood brain barrier, that may be useful for drug delivery studies, and for the formulation of new therapeutic strategies, to be used for the treatment of neurological diseases

Engineering approaches in siRNA delivery

siRNAs are very potent drug molecules, able to silence genes involved in pathologies development. siRNAs have virtually an unlimited therapeutic potential, particularly for the treatment of inflammatory diseases. However, their use in clinical practice is limited because of their unfavorable properties to interact and not to degrade in physiological environments. In particular they are large macromolecules, negatively charged, which undergo rapid degradation by plasmatic enzymes, are subject to fast renal clearance/hepatic sequestration, and can hardly cross cellular membranes. These aspects seriously impair siRNAs as therapeutics. As in all the other fields of science, siRNAs management can be advantaged by physical-mathematical descriptions (modeling) in order to clarify the involved phenomena from the preparative step of dosage systems to the description of drug-body interactions, which allows improving the design of delivery systems/processes/therapies. This review analyzes a few mathematical modeling approaches currently adopted to describe the siRNAs delivery, the main procedures in siRNAs vectors\ue2\u80\u99 production processes and siRNAs vectors\ue2\u80\u99 release from hydrogels, and the modeling of pharmacokinetics of siRNAs vectors. Furthermore, the use of physical models to study the siRNAs vectors\ue2\u80\u99 fate in blood stream and in the tissues is presented. The general view depicts a framework maybe not yet usable in therapeutics, but with promising possibilities for forthcoming applications

Predictors of length of hospital stay among older adults admitted to acute care wards: a multicentre observational study

Reduction in length of hospital stay (LOS) is considered as a potential strategy to optimize resource consumption and reduce health care costs. We analysed predictors of increased LOS among older patients admitted to acute care wards according to type of admission (through the Emergency Room [ER] or elective)

Bile modulates secretion of incretins and insulin: a study on human extrahepatic cholestasis

OBJECTIVE: changes in bile flow after bariatric surgery may beneficially modulate secretion of insulin and incretins leading to diabetes remission. However, the exact mechanism(s) involved are still unclear. We here propose an alternative method to investigate the relation between alterations in physiological bile flow and insulin and incretin secretion by studying the changes in gut-pancreatic function in extrahepatic cholestasis in non-diabetic humans. METHODS: to pursue this aim, 58 non-diabetic patients with recent diagnosis of periampullary tumors underwent an OGTT, and a subgroup of 16 patients also underwent 4-hour mixed meal tests and hyperinsulinemic euglycemic clamps. RESULTS: The analysis of the entire cohort revealed strong inverse correlation between total bilirubin levels and insulinogenic index. When subjects were divided on the basis of bilirubin levels, used as marker of altered bile flow, subjects with high bilirubin levels displayed inferior glucose control and decreased insulin secretion during OGTT. Altered bile flow elicited a markedly greater increase in glucagon and GLP-1 secretion at fasting state and following the meal both glucagon and GLP-1 levels remained increased over time. Conversely, GIP levels were comparable at the fasting state, while the increase following meal ingestion was significantly blunted in H-Bil. We revealed strong correlations between total bilirubin and glucagon and GLP-1 levels. CONCLUSIONS: Our findings suggest that acute extrahepatic cholestasis determines a significant impairment in the entero-endocrine gut-pancreatic secretory function. The altered bile flow might determine a direct deleterious effect on beta cell function perhaps mediated by the impairment of incretin hormone function