Role of proteoglycans and glycosaminoglycans in tumorigenesis

La variabilità strutturale e funzionale dei proteoglicani (PGs) è alla base della loro fondamentale importanza biologica nei processi di sviluppo, nella vita adulta e in diverse condizioni patologiche come il cancro. I proteoglicani sono coinvolti in diversi processi cellulari grazie alla loro capacità di legare, modulandone l’attività, diversi fattori del crescita. Tra tutti i proteoglicani, i glipicani (GPCs) sono indicati sia come molecole tumore-associate sia come soppressori tumorali e la loro funzione sembra essere fortemente collegata con il fenotipo di tumore. I nostri studi si sono concentrati su un particolare glipicano, il glipicano 5 (GPC5), in un particolare fenotipo di tumore delle parti molli, l’osteosarcoma umano. I nostri dati supportano l’ipotesi che il glipicano 5 possa essere considerato come un soppressore della tumorigenesi in questi particolari fenotipi tumorali, agendo sia sulla proliferazione delle cellule tumorali sia sulla loro motilità. I glicosaminoglicani (GAGs) partecipano a molti dei processi biologici influenzati dai proteoglicani e la loro complessità funzionale è in parte correlata con le loro caratteristiche strutturali e chimiche. Per questa ragione abbiamo ritenuto opportuno studiare approfonditamente due particolari strutture glicosaminoglicaniche, catene di eparan solfato ad alta solfatazione e catene di cheratan solfato caratterizzate da una particolare struttura chimica. Questi studi sono stati condotti per mezzo della tecnica degli anticorpi monoclonali. Il nostro scopo ultimo è quello di tracciare la correlazione esistente tra struttura chimica di queste molecole e loro distribuzione nei tessuti umani. Questi studi rappresentano passi importanti nella comprensione profonda della relazione tra struttura e funzione dei proteoglicani che potrebbe rendere queste molecole degli interessanti bersagli terapeutici in diverse condizioni patologiche.Structural-functional diversity of proteoglycans (PGs) is the pivot of their biological importance during the embryonic development, in the adult and also in pathological conditions such as cancer. PGs may be involved in different cell behavior by binding and modulating a broad spectrum of growth factors. In particular, among PGs, glypicans (GPCs) are listed as both tumor-inhibitors and promoters and seems to be closely tumor-type specific. Our study on glypican 5 (GPC5) in a particular soft tissue sarcoma, human osteosarcoma, support the hypothesis that this molecule may be considered as a tumor-suppressor, inhibiting cancer cell growth and motility. Glycosaminoglycans (GAG) participate to many of the PG biological processes and their functional complexity is in part due to their structural and chemical characteristics. For this reason we decided to investigate two particular GAG chains, highly sulfated heparin sulfate moieties and a specific keratan sulfate structure, using the monoclonal antibody technique. Our goal is to understand the correlation between the chemical structures of these moieties and their distribution in the human adult tissues. These may be the first steps in the comprehension of the structural-functional relationship of PGs that could be considered as an attractive target for therapeutic intervention in different pathological conditions

The Utilisation Of Thermal Methods For The Screening Of Three Component Co-amorphous Systems

Indomethacin and piroxicam are known to become amorphous upon rapid cooling after melting. Multiple three-component co-amorphous mixtures have been produced via a melt quench method to evaluate the influence of the third component on stability and physical properties on the co-amorphous mixture of piroxicam and indomethacin. These have then been tested using thermal gravimetric analysis, differential scan calorimetry and hot stage microscopy. Results have confirmed the creation of a three-component co-amorphous system

Micro-photogrammetry as a tool for characterisation of dissolution behaviour of pharmaceutical dosage forms

The aim of this project is to develop an innovative analytical technique for the 3D chemical mapping of pharmaceutical dosage forms, via simultaneous topographic characterisation and dissolution analysis of solid drug delivery systems. The developed apparatus was optimised using the results obtained from the application of micro-photogrammetry to the analysis of commercial solid drug delivery systems. Results of the application of the developed method on the dissolution of a sugarcoated tablet sample are here presented

The Use Of XRPD And ATR-FTIR In The Screening Of Three Component Co-amorphous Systems Created Via A Melt-Quench Method.

Multiple three-component co-amorphous systems have been created via a melt- quench method. This has been confirmed by a single glass transition in the individual DSC traces and then reinforced by X-ray diffraction data, which show an absence of crystalline material. ATR-FTIR has also been used to show a change in bonding vibrations between the physical mixtures and co-amorphous samples. The change in bonding vibrations indicates different molecular interactions within the co-amorphous materials

Indigenous enzymes and leukocyte in sheep milk are markers of health status and physiology of the mammary gland

Plasmin, plasminogen and plasminogen activator in ewe bulk milk were not significantly affected by stage of lactation probably as a consequence of the good health of the ewe udders throughout lactation as indicated by SCC which never exceeded 600,000 cells/mL. Elastase content increased significantly during lactation whereas cathepsin showed the highest content in mid lactation. Changes in macrophages and neutrophyls levels in ewe bulk milk during lactation were also investigated. Macrophages minimally contributed to leukocyte cell count in milk and had the highest levels at the beginning of lactation. An opposite trend was recorded for polymorphonuclear leukocytes (PMNL) that increased throughout lactation showing the highest value in late lactation. The increase of PMNL percentage and elastase content in milk, in spite of relatively low SCC, suggests that PMNL and elastase underwent a physiological increase associated to the remodelling of mammary gland in late lactation

Evidence and consensus on technical aspects of embryo transfer

BACKGROUND Ultrasound-guided embryo transfer (US-GET) is a widely performed procedure, but standards for the best practice are not available. OBJECTIVE AND RATIONALE This document aims to provide an overview of technical aspects of US-GET after considering the published data and including the preparation for the embryo transfer (ET) procedure, the actual procedure, the post-procedure care, associated pathologies, complications and risks, quality assurance and practitioners' performance. SEARCH METHODS A literature search for evidence on key aspects of the ET procedure was carried out from database inception to November 2021. Selected papers (n = 359) relevant to the topic were analysed by the authors. The following key points were considered in the papers: whether ultrasound (US) practice standards were explained, to what extent the ET technique was described and whether complications or incidents and how to prevent such events were reported. In the end, 89 papers could be used to support the recommendations in this document, which focused on transabdominal US-GET. OUTCOMES The relevant papers found in the literature search were included in the current document and described according to the topic in three main sections: requirements and preparations prior to ET, the ET procedure and training and competence for ET. Recommendations are provided on preparations prior to ET, equipment and materials, ET technique, possible risks and complications, training and competence. Specific aspects of the laboratory procedures are covered, in particular the different loading techniques and their potential impact on the final outcomes. Potential future developments and research priorities regarding the ET technique are also outlined. LIMITATIONS, REASONS FOR CAUTION Many topics were not covered in the literature review and some recommendations were based on expert opinions and are not necessarily evidence based. WIDER IMPLICATIONS ET is the last procedural step in an ART treatment and is a crucial step towards achieving a pregnancy and live birth. The current paper set out to bring together the recent developments considering all aspects of ET, especially emphasizing US quality imaging. There are still many questions needing answers, and these can be subject of future research. STUDY FUNDING/COMPETING INTEREST(S) No funding. A.D.A. has received royalties from CRC Press and personal honorarium from Cook, Ferring and Cooper Surgical. The other co-authors have no conflicts of interest to declare that are relevant to the content of this article.Peer reviewe

A Multiple-Plasticity Spiking Neural Network Embedded in a Closed-Loop Control System to Model Cerebellar Pathologies

The cerebellum plays a crucial role in sensorimotor control and cerebellar disorders compromise adaptation and learning of motor responses. However, the link between alterations at network level and cerebellar dysfunction is still unclear. In principle, this understanding would benefit of the development of an artificial system embedding the salient neuronal and plastic properties of the cerebellum and operating in closed-loop. To this aim, we have exploited a realistic spiking computational model of the cerebellum to analyze the network correlates of cerebellar impairment. The model was modified to reproduce three different damages of the cerebellar cortex: (i) a loss of the main output neurons (Purkinje Cells), (ii) a lesion to the main cerebellar afferents (Mossy Fibers), and (iii) a damage to a major mechanism of synaptic plasticity (Long Term Depression). The modified network models were challenged with an Eye-Blink Classical Conditioning test, a standard learning paradigm used to evaluate cerebellar impairment, in which the outcome was compared to reference results obtained in human or animal experiments. In all cases, the model reproduced the partial and delayed conditioning typical of the pathologies, indicating that an intact cerebellar cortex functionality is required to accelerate learning by transferring acquired information to the cerebellar nuclei. Interestingly, depending on the type of lesion, the redistribution of synaptic plasticity and response timing varied greatly generating specific adaptation patterns. Thus, not only the present work extends the generalization capabilities of the cerebellar spiking model to pathological cases, but also predicts how changes at the neuronal level are distributed across the network, making it usable to infer cerebellar circuit alterations occurring in cerebellar pathologies

Automatic Setting Procedure for Exoskeleton-Assisted Overground Gait: Proof of Concept on Stroke Population

Stroke-related locomotor impairments are often associated with abnormal timing and intensity of recruitment of the affected and non-affected lower limb muscles. Restoring the proper lower limbs muscles activation is a key factor to facilitate recovery of gait capacity and performance, and to reduce maladaptive plasticity. Ekso is a wearable powered exoskeleton robot able to support over-ground gait training. The user controls the exoskeleton by triggering each single step during the gait cycle. The fine-tuning of the exoskeleton control system is crucial-it is set according to the residual functional abilities of the patient, and it needs to ensure lower limbs powered gait to be the most physiological as possible. This work focuses on the definition of an automatic calibration procedure able to detect the best Ekso setting for each patient. EMG activity has been recorded from Tibialis Anterior, Soleus, Rectus Femoris, and Semitendinosus muscles in a group of 7 healthy controls and 13 neurological patients. EMG signals have been processed so to obtain muscles activation patterns. The mean muscular activation pattern derived from the controls cohort has been set as reference. The developed automatic calibration procedure requires the patient to perform overground walking trials supported by the exoskeleton while changing parameters setting. The Gait Metric index is calculated for each trial, where the closer the performance is to the normative muscular activation pattern, in terms of both relative amplitude and timing, the higher the Gait Metric index is. The trial with the best Gait Metric index corresponds to the best parameters set. It has to be noted that the automatic computational calibration procedure is based on the same number of overground walking trials, and the same experimental set-up as in the current manual calibration procedure. The proposed approach allows supporting the rehabilitation team in the setting procedure. It has been demonstrated to be robust, and to be in agreement with the current gold standard (i.e., manual calibration performed by an expert engineer). The use of a graphical user interface is a promising tool for the effective use of an automatic procedure in a clinical context

Progettazione e Quality of Life nella presa in carico di persone con disabilità intellettive: l’agricoltura sociale come nuovo spazio e tempo generativo

Starting from the theoretical framework of Quality of Life (Giaconi, 2015b; Schalock, Verdugo Alonso, 2006, 2002), the paper explores the continuity between the results of two assessment scales (POS and SIS) in a case study. The case concerns the initial condition of a young person with intellectual disabilities involved in a social agriculture project, called “Tuttincampo”. The project involves the synergistic collaboration of an inter-institutional network composed of public and private entities, and is aimed at testing a new format of social and labour integration, with the goal of offering a viable alternative to classic rehabilitation day centres. In detail, we want to explore the first useful data to support the project pathway aimed at deepening the practices and methodologies to support the social and labour inclusion of people with intellectual disabilities.Starting from the theoretical framework of Quality of Life (Giaconi, 2015b; Schalock, Verdugo Alonso, 2006, 2002), the paper explores the continuity between the results of two assessment scales (POS and SIS) in a case study. The case con­cerns the initial condition of a young person with intellectual disabilities involved in a social agriculture project, called “Tuttincampo”. The project involves the synergistic collaboration of an inter­institutional network composed of public and private entities, and is aimed at testing a new format of social and labour integration, with the goal of offering a viable alternative to classic rehabilitation day centres. In detail, we want to explore the first useful data to support the project pathway aimed at deepening the practices and methodologies to support the social and labour inclusion of people with intellectual disabilities

VALUTAZIONE DEL CALCIO INTRACELLULARE IN PIASTINE DI CANI SANI ED AFFETTI DA LEISHMANIOSI

Le piastrine rappresentano gli elementi cellulari fondamentali per il processo di regolazione dell’emostasi. La loro interazione con l’endotelio vasale riveste un ruolo fondamentale sia nella fisiologia che nella fisiopatologia della coagulazione. fisiologicamente, infatti, le piastrine hanno la capacità di aderire alla parete del vaso solo dopo attivazione da parte dell’endotelio vasale. In condizioni patologiche, come quelle causate dalla rottura di una placca aterosclerotica, le piastrine aderiscono all’endotelio, si attivano e reclutano numerose altre piastrine per la formazione del tappo piastrinico. In medicina veterinaria, diversamente dalla medicina umana, la conoscenza dei meccanismi fisiologici specifici alla base di tali processi è piuttosto scarsa. Le piastrine subiscono fenomeni di attivazione, fra i quali è possibile menzionare, lo shape change, l’attivazione del recettore per il fibrinogeno che causa l’aggregazione delle piastrine, il rilascio del contenuto dei granuli e la produzione di trombossano A2 (Gachet C. et al., 1996, Mills D.C.B., 1996). Tali fenomeni sono mediati da modificazioni del calcio intracellulare che origina dal rilascio di calcio dagli store intracellulari e dall’entrata di calcio dal mezzo extracellulare (Rink T.J. et al., 1985, Rink T.J. et al., 1990). D’altro canto però, sono note numerose patologie nel cane, tra cui forme di Leishmaniosi eo Ehrlichiosi, che comportano seria compromissione della funzione coagulativa (Harrus S. et al., 1996; Weiss D.J. et al., 1995). La leishmaniosi, in particolare, è una zoonosi largamente diffusa nelle zone costiere del Mediterraneo caratterizzata da una alterazione della coagulazione spesso asintomatica che si inserisce in un quadro sintomatologico estremamente vario. In precedenti ricerche abbiamo mostrato che, in corso di leishmaniosi, si verifica un danno piastrinico di entità variabile che, peraltro, non trova completa risoluzione con le terapie normalmente utilizzate in corso di tale patologia. I meccanismi attraverso i quali l’aggregazione piastrinica risulta alterata in corso di leishmaniosi non sono ancora chiari. Per tale motivo, l’obiettivo di questo studio è stato quello di valutare le variazioni di calcio nelle piastrine di cani sani ed affetti da leishmaniosi allo scopo di raggiungere una migliore comprensione dei meccanismi responsabili di alterazioni dell’emostasi in corso di tale patologia