Search results for: Della Rovere Federica

Authors: Engy Shokry, Giancarlo La Marca, Maria Luisa Della Bona

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Newborn screening for Congenital Adrenal Hyperplasia (CAH) relies on quantification of 17α-hydroxyprogesterone using enzyme immunoassays. These assays, in spite of being rapid, readily available and easy to perform, its reliability was found questionable due to lack of selectivity and specificity resulting in large number of false-positives, consequently family anxiety and associated hospitalization costs. To improve specificity of conventional 17α-hydroxyprogesterone screening which may experience false transient elevation in preterm, low birth weight or acutely ill neonates, steroid profiling by LC-MS/MS as a second-tier test was implemented. Unlike the previously applied LC-MS/MS methods, with the disadvantage of requiring a relatively high number of blood drops. Since newborn screening tests are increasing, it is necessary to minimize the sample volume requirement to make the maximum use of blood samples collected on filter paper. The proposed new method requires just one 3.2 mm dried blood spot (DBS) punch. Extraction was done using methanol: water: formic acid (90:10:0.1, v/v/v) containing deuterium labelled internal standards. Extracts were evaporated and reconstituted in 10 % acetone in water. Column switching strategy for on-line sample clean-up was applied to improve the chromatographic run. The first separative step retained the investigated steroids and passed through the majority of high molecular weight impurities. After the valve switching, the investigated steroids are back flushed from the POROS® column onto the analytical column and separated using gradient elution. Found quantitation limits were 5, 10 and 50 nmol/L for 17α-hydroxyprogesterone, androstenedione and cortisol respectively with mean recoveries of between 98.31-103.24 % and intra-/ inter-assay CV% < 10 % except at LLOQ. The method was validated using standard addition calibration and isotope dilution strategies. Reference ranges were determined by analysing samples from 896 infants of various ages at the time of sample collection. The method was also applied on patients with confirmed CAH. Our method represents an attractive combination of low sample volume requirement, minimal sample preparation time without derivatization and quick chromatography (5 min). The three steroid profile and the concentration ratios (17OHP + androstenedione/cortisol) allowed better screening outcomes of CAH reducing false positives, associated costs and anxiety.

Keywords: congenital adrenal hyperplasia (CAH), 17α-hydroxyprogesterone, androstenedione, cortisol, LC-MS/MS

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Authors: Clara Tramuta, Floris Irene, Daniela Manila Bianchi, Monica Pitti, Giulia Federica Cazzaniga, Lucia Decastelli

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This work presents the results obtained by the Regional Reference Centre for Salmonella Typing (CeRTiS) in a retrospective study aimed to investigate, through Pulsed-field Gel Electrophoresis (PFGE) analysis, the genetic relatedness of emerging Salmonella serotypes of human origin circulating in North-West of Italy. Furthermore, the goal of this work was to create a Regional database to facilitate foodborne outbreak investigation and to monitor them at an earlier stage. A total of 112 strains, isolated from 2016 to 2018 in hospital laboratories, were included in this study. The isolates were previously identified as Salmonella according to standard microbiological techniques and serotyping was performed according to ISO 6579-3 and the Kaufmann-White scheme using O and H antisera (Statens Serum Institut®). All strains were characterized by PFGE: analysis was conducted according to a standardized PulseNet protocol. The restriction enzyme XbaI was used to generate several distinguishable genomic fragments on the agarose gel. PFGE was performed on a CHEF Mapper system, separating large fragments and generating comparable genetic patterns. The agarose gel was then stained with GelRed® and photographed under ultraviolet transillumination. The PFGE patterns obtained from the 112 strains were compared using Bionumerics version 7.6 software with the Dice coefficient with 2% band tolerance and 2% optimization. For each serotype, the data obtained with the PFGE were compared according to the geographical origin and the year in which they were isolated. Salmonella strains were identified as follow: S. Derby n. 34; S. Infantis n. 38; S. Napoli n. 40. All the isolates had appreciable restricted digestion patterns ranging from approximately 40 to 1100 kb. In general, a fairly heterogeneous distribution of pulsotypes has emerged in the different provinces. Cluster analysis indicated high genetic similarity (≥ 83%) among strains of S. Derby (n. 30; 88%), S. Infantis (n. 36; 95%) and S. Napoli (n. 38; 95%) circulating in north-western Italy. The study underlines the genomic similarities shared by the emerging Salmonella strains in Northwest Italy and allowed to create a database to detect outbreaks in an early stage. Therefore, the results confirmed that PFGE is a powerful and discriminatory tool to investigate the genetic relationships among strains in order to monitoring and control Salmonellosis outbreak spread. Pulsed-field gel electrophoresis (PFGE) still represents one of the most suitable approaches to characterize strains, in particular for the laboratories for which NGS techniques are not available.

Keywords: emerging Salmonella serotypes, genetic characterization, human strains, PFGE

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Authors: Federica Marangio

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This study looks at the triangle of knowledge: research-education-innovation as growth engine of an inclusive and sustainable society, where the research is the strategic process which allows the acquisition of knowledge, innovation appraises the knowledge acquired and the education is the enabling factor of the human capital to create entrepreneurial capital. Where does Italy and China stand in the global geography of innovation? Europe is calling on a smart, inclusive and sustainable growth through a specializing process that looks at the social and economic challenges, able to understand the characteristics of specific geographic areas. It is easily questionable why it is not as simple as it looks to come up with entrepreneurial ideas in all the geographic areas. Seen that the technology plus the human capital should be the means through which is possible to innovate and contribute to the boost of innovation culture, then the young educated people can be seen as the society changing agents and it becomes clear the importance of investigating the skills and competencies that lead to innovation. By starting innovation-based activities, other countries on an international level, are able now to be part of an healthy innovative ecosystem which is the result of a strong growth policy which enables innovation. Analyzing the geography of the innovation on a global scale, comes to light that the innovative entrepreneurship is the process which portrays the competitiveness of the regions in the knowledge-based economy as strategic process able to match intellectual capital and market opportunities. The level of innovative entrepreneurship is not only the result of the endogenous growth ability of the enterprises, but also by significant relations with other enterprises, universities, other centers of education and institutions. To obtain more innovative entrepreneurship is necessary to stimulate more synergy between all these territory actors in order to create, access and value existing and new knowledge ready to be disseminate. This study focuses on individual’s lived experience and the researcher believed that she can’t understand the human actions without understanding the meaning that they attribute to their thoughts, feelings, beliefs and so given she needed to understand the deeper perspectives captured through face-to face interaction. A case study approach will contribute to the betterment of knowledge in this field. This case study will represent a picture of the innovative ecosystem and the entrepreneurial mindset as a key ingredient of endogenous growth and a must for sustainable local and regional development and social cohesion. The case study will be realized analyzing two Chinese companies. A structured set of questions will be asked in order to gain details on what generated success or failure in the different situations with the past and at the moment of the research. Everything will be recorded not to lose important information during the transcription phase. While this work is not geared toward testing a priori hypotheses, it is nevertheless useful to examine whether the projects undertaken by the companies, were stimulated by enabling factors that, as result, enhanced or hampered the local innovation culture.

Keywords: Entrepreneurship, education, geography of innovation, education.

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Authors: Mark Opmeer

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The fruitful relationship between cultural heritage and digital technology is evident. Due to the development of user-friendly software, an increasing amount of heritage scholars use ict for their research activities. As a result, the implementation of information technology for heritage planning has become a research objective in itself. During the last decades, we have witnessed a growing debate and literature about the importance of computer technologies for the field of cultural heritage and ecotourism. Indeed, implementing digital technology in support of these domains can be very fruitful for one’s research practice. However, due to the rapid development of new software scholars may find it challenging to use these innovations in an appropriate way. As such, this contribution seeks to explore the interplay between geospatial technologies (geo-ict), civic engagement and cultural heritage and tourism. In this article, we discuss our findings on the use of geo-ict in support of civic participation, cultural heritage and sustainable tourism development in the southern Italian district of Brindisi. In the city of Latiano, three workshops were organized that involved local members of the community to distinguish and discuss interesting points of interests (POI’s) which represent the cultural significance and identity of the area. During the first workshop, a so called mappa della comunità was created on a touch table with collaborative mapping software, that allowed the participators to highlight potential destinations for tourist purposes. Furthermore, two heritage-based itineraries along a selection of identified POI’s was created to make the region attractive for recreants and tourists. These heritage-based itineraries reflect the communities’ ideas about the cultural identity of the region. Both trails were subsequently implemented in a dedicated mobile application (app) and was evaluated using a mixed-method approach with the members of the community during the second workshop. In the final workshop, the findings of the collaboration, the heritage trails and the app was evaluated with all participants. Based on our conclusions, we argue that geospatial technologies have a significant potential for involving local communities in heritage planning and tourism development. The participants of the workshops found it increasingly engaging to share their ideas and knowledge using the digital map of the touch table. Secondly, the use of a mobile application as instrument to test the heritage-based itineraries in the field was broadly considered as fun and beneficial for enhancing community awareness and participation in local heritage. The app furthermore stimulated the communities’ awareness of the added value of geospatial technologies for sustainable tourism development in the area. We conclude this article with a number of recommendations in order to provide a best practice for organizing heritage workshops with similar objectives.

Keywords: civic engagement, geospatial technologies, tourism development, cultural heritage

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Authors: Federica Ceccoli

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Italy is undergoing one of its largest migratory waves, and Italian schools are reporting the highest numbers of multilingual students coming from immigrant families and speaking minority languages. For these pupils, who have not perfectly acquired their mother tongue yet, learning a second language may represent a burden on their linguistic development and may have some repercussions on their school performances and relational skills. These are some of the reasons why they have turned out to be those who have the worst grades and the highest school drop-out rates. However, despite these negative outcomes, it has been demonstrated that multilingual immigrant students frequently act as translators or language brokers for their peers or family members who do not speak Italian fluently. This activity has been defined as Child Language Brokering (hereinafter CLB) and it has become a common practice especially in minority communities as immigrants’ children often learn the host language much more quickly than their parents, thus contributing to their family life by acting as language and cultural mediators. This presentation aims to analyse the data collected by a research carried out during the school year 2014-2015 in the province of Ravenna, in the Northern Italian region of Emilia-Romagna, among 126 immigrant students attending junior high schools. The purpose of the study was to analyse by means of a structured questionnaire whether multilingualism matched with language brokering experiences or not and to examine the perspectives of those students who reported having acted as translators using their linguistic knowledge to help people understand each other. The questionnaire consisted of 34 items roughly divided into 2 sections. The first section required multilingual students to provide personal details like their date and place of birth, as well as details about their families (number of siblings, parents’ jobs). In the second section, they were asked about the languages spoken in their families as well as their language brokering experience. The in-depth questionnaire sought to investigate a wide variety of brokering issues such as frequency and purpose of the activity, where, when and which documents young language brokers translate and how they feel about this practice. The results have demonstrated that CLB is a very common practice among immigrants’ children living in Ravenna and almost all students reported positive feelings when asked about their brokering experience with their families and also at school. In line with previous studies, responses to the questionnaire item regarding the people they brokered for revealed that the category ranking first is parents. Similarly, language-brokering activities tend to occur most often at home and the documents they translate the most (either orally or in writing) are notes from teachers. Such positive feelings towards this activity together with the evidence that it occurs very often in schools have laid the foundation for further projects on how this common practice may be valued and used to strengthen the linguistic skills of these multilingual immigrant students and thus their school performances.

Keywords: immigration, language brokering, multilingualism, students' points of view

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Authors: Beste Ozcan, Valerio Sperati, Laura Romano, Tania Moretta, Simone Scaffaro, Noemi Faedda, Federica Giovannone, Carla Sogos, Vincenzo Guidetti, Gianluca Baldassarre

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+me is an experimental interactive toy with the appearance of a soft, pillow-like, panda. Shape and consistency are designed to arise emotional attachment in young children: a child can wear it around his/her neck and treat it as a companion (i.e. a transitional object). When caressed on paws or head, the panda emits appealing, interesting outputs like colored lights or amusing sounds, thanks to embedded electronics. Such sensory patterns can be modified through a wirelessly connected tablet: by this, an adult caregiver can adapt +me responses to a child's reactions or requests, for example, changing the light hue or the type of sound. The toy control is therefore shared, as it depends on both the child (who handles the panda) and the adult (who manages the tablet and mediates the sensory input-output contingencies). These features make +me a potential tool for therapy with children with Neurodevelopmental Disorders (ND), characterized by impairments in the social area, like Autism Spectrum Disorders (ASD) and Language Disorders (LD): as a proposal, the toy could be used together with a therapist, in rehabilitative play activities aimed at encouraging simple social interactions and reinforcing basic relational and communication skills. +me was tested in two pilot experiments, the first one involving 15 Typically Developed (TD) children aged in 8-34 months, the second one involving 7 children with ASD, and 7 with LD, aged in 30-48 months. In both studies a researcher/caregiver, during a one-to-one, ten-minute activity plays with the panda and encourages the child to do the same. The purpose of both studies was to ascertain the general acceptability of the device as an interesting toy that is an object able to capture the child's attention and to maintain a high motivation to interact with it and with the adult. Behavioral indexes for estimating the interplay between the child, +me and caregiver were rated from the video recording of the experimental sessions. Preliminary results show how -on average- participants from 3 groups exhibit a good engagement: they touch, caress, explore the panda and show enjoyment when they manage to trigger luminous and sound responses. During the experiments, children tend to imitate the caregiver's actions on +me, often looking (and smiling) at him/her. Interesting behavioral differences between TD, ASD, and LD groups are scored: for example, ASD participants produce a fewer number of smiles both to panda and to a caregiver with respect to TD group, while LD scores stand between ASD and TD subjects. These preliminary observations suggest that the interactive toy +me is able to raise and maintain the interest of toddlers and therefore it can be reasonably used as a supporting tool during therapy, to stimulate pivotal social skills as imitation, turn-taking, eye contact, and social smiles. Interestingly, the young age of participants, along with the behavioral differences between groups, seem to suggest a further potential use of the device: a tool for early differential diagnosis (the average age of a child

Keywords: autism spectrum disorders, interactive toy, social interaction, therapy, transitional wearable companion

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Authors: Tramuta Clara, Masotti Chiara, Pitti Monica, Adriano Daniela, Battistini Roberta, Serraca Laura, Decastelli Lucia

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Oysters are considered filter organisms, and their raw consumption may increase health risks for consumers: it is often associated with outbreaks of gastroenteritis or enteric illnesses. Most of these foodborne diseases are caused by Vibrio strains, enteric pathogens also involved in the diffusion of genetic determinants of antibiotic resistance and their entrance along the food chain. The European Food Safety Authority (EFSA), during the European Union report on antimicrobial resistance in 2017, focused the attention about the role of food as a possible carrier of antibiotic-resistant bacteria or antibiotic-resistance genes that determine health risks for humans. This study wants to determine antibiotic resistance and antibiotic-resistance genes in Vibrio spp. isolated from Crassostrea gigas oysters collected in the Golfo della Spezia (Liguria, Italy). A total of 47 Vibrio spp. strains were isolated (ISO21872-2:2017) during the summer of 2021 from oysters of Crassostrea gigas. The strains were identified by MALDI-TOF (Bruker, Germany) mass spectrometry and tested for antibiotic susceptibility using a broth microdiluition method (ISO20776-1:2019) using Sensititre EUVSEC plates (Thermo-Fisher Scientific) to obtain the Minimum Inhibitory Concentration (MIC). The strains were tested with PCR-based biomolecular methods, according to previous works, to define the presence of 23 resistance genes of the main classes of antibiotics used in human and veterinary medicine: tet (B), tet (C), tet (D), tet (A), tet (E), tet (G ), tet (K), tet (L), tet (M), tet (O), tet (S) (tetracycline resistance); blaCTX-M, blaTEM, blaOXA, blaSHV (β-lactam resistance); mcr-1 and mcr-2 (colistin resistance); qnrA, qnrB, and qnrS (quinolone resistance); sul1, sul2 and sul3 (sulfonamide resistance). Six different species have been identified: V. alginolyticus 34% (n=16), V. harveyi 28% (n=13), V. fortis 15% (n=7), V. pelagius 8% (n=4), V. parahaemolyticus 11% (n=5) e V. chagasii 4% (n=2). The PCR assays showed the presence of the blaTEM gene on 40% of the strains (n=19). All the other genes were not detected, except for a V. alginolyticus positive for anrS gene. The broth microdiluition method results showed an high level of resistance for ciprofloxacin (62%; n=29), ampicillin (47%; n=22), and colistin (49%; n=23). Furthermore, 32% (n=15) of strains can be considered multiresistant bacteria for the simultaneous presence of resistance for three different antibiotic classes. Susceptibility towards meropenem, azithromycin, gentamicin, ceftazidime, cefotaxime, chloramphenicol, tetracycline and sulphamethoxazole reached 100%. The Vibrio species identified in this study are widespread in marine environments and can cause gastrointerstinal infections after the ingestion of raw fish products and bivalve molluscs. The level of resistance to antibiotics such as ampicillin, ciprofloxacin and colistin can be connected to anthropic factors (industrial, agricultural and domestic wastes) that promote the spread of resistance to these antibiotics. It can be also observed a strong correlation between phenotypic (resistant MIC) and genotypic (positive blaTEM gene) resistance for ampicillin on the same strains, probably due to the transfer of genetic material between bacterial strains. Consumption of raw bivalve molluscs can represent a risk for consumers heath due to the potentially presence of foodborne pathogens, highly resistant to different antibiotics and source of transferable antibiotic-resistant genes.

Keywords: vibrio species, blaTEM genes, antimicrobial resistance, PCR

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Authors: Federica Verna, Alessandra Pautasso, Maria Caramelli, Cristiana Maurella, Walter Mignone, Cristina Casalone

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Mosquito-Borne Diseases (MBDs) are dangerously increasing in prevalence, geographical distribution and severity, representing an emerging threat for both humans and animals. Interaction between multiple disciplines is needed for an effective early warning, surveillance and control of MBDs, according to the One Health concept. This work reports the integrated surveillance system enforced by IZSPLV in Piedmont, Liguria and Valle d’Aosta regions (Northwestern Italy) in order to control MDBs spread. Veterinary services and local human health authority are involved in an information network, to connect the surveillance of human clinical cases with entomological surveillance and veterinary monitoring in order to implement control measures in case of outbreak. A systematic entomological surveillance is carried out during the vector season using mosquitoes traps located in sites selected according to risk factors. Collected mosquitoes are counted, identified to species level by morphological standard classification keys and pooled by collection site, date and species with a maximum of 100 individuals. Pools are analyzed, after RNA extraction, by Real Time RT-PCR distinctive for West Nile Virus (WNV) Lineage 1 and Lineage 2, Real Time RT-PCR USUTU virus (USUV) and a traditional flavivirus End-point RT-PCR. Positive pools are sequenced and the related sequences employed to perform a basic local alignment search tool (BLAST) in the GenBank library. Positive samples are sent to the National Reference Centre for Animal Exotic Diseases (CESME, Teramo) for confirmation. With particular reference to WNV, after the confirmation, as provided by national legislation, control measures involving both local veterinary and human health services are activated: equine sera are randomly sampled within a 4 km radius from the positive collection sites and tested with ELISA kit and WNV NAT screening of blood donors is introduced. This surveillance network allowed to detect since 2011 USUV circulation in this area of Italy. WNV was detected in Piedmont and Liguria for the first time in 2014 in mosquitoes. During the 2015 vector season, we observed the expansion of its activity in Piedmont. The virus was detected in almost all Provinces both in mosquitoes (6 pools) and animals (19 equine sera, 4 birds). No blood bag tested resulted infected. The first neuroinvasive human case occurred too. Competent authorities should be aware of a potentially increased risk of MBDs activity during the 2016 vector season. This work shows that this surveillance network allowed to early detect the presence of MBDs in humans and animals, and provided useful information to public authorities, in order to apply control measures. Finally, an additional value of our diagnostic protocol is the ability to detect all viruses belonging to the Flaviviridae family, considering the emergence caused by other Flaviviruses in humans such as the recent Zika virus infection in South America. Italy has climatic and environmental features conducive to Zika virus transmission, the competent vector and many travellers from Brazil reported every year.

Keywords: integrated surveillance, mosquito borne disease, West Nile virus, Zika virus

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Authors: Francois Brambati, Daniele Ruscio, Federica Biassoni, Rebecca Hueting, Alessandra Tedeschi

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This research, in the frame of social acceptance of renewable energies and community-based production and consumption models, aims at (1) supporting a data-driven approachable to dealing with climate change and (2) identifying & quantifying the psycho-sociological dimensions and factors that could support the transition from a technology-driven approach to a consumer-driven approach throughout the emerging “prosumer business models.” In addition to the existing Social Acceptance dimensions, this research tries to identify a purely individual psychological fourth dimension to understand processes and factors underling individual acceptance and adoption of renewable energy business models, realizing a Prosumer Acceptance Index. Questionnaire data collection has been performed throughout an online survey platform, combining standardized and ad-hoc questions adapted for the research purposes. To identify the main factors (individual/social) influencing the relation with renewable energy technology (RET) adoption, a Factorial Analysis has been conducted to identify the latent variables that are related to each other, revealing 5 latent psychological factors: Factor 1. Concern about environmental issues: global environmental issues awareness, strong beliefs and pro-environmental attitudes rising concern on environmental issues. Factor 2. Interest in energy sharing: attentiveness to solutions for local community’s collective consumption, to reduce individual environmental impact, sustainably improve the local community, and sell extra energy to the general electricity grid. Factor 3. Concern on climate change: environmental issues consequences on climate change awareness, especially on a global scale level, developing pro-environmental attitudes on global climate change course and sensitivity about behaviours aimed at mitigating such human impact. Factor 4. Social influence: social support seeking from peers. With RET, advice from significant others is looked for internalizing common perceived social norms of the national/geographical region. Factor 5. Impact on bill cost: inclination to adopt a RET when economic incentives from the behaviour perception affect the decision-making process could result in less expensive or unvaried bills. Linear regression has been conducted to identify and quantify the factors that could better predict behavioural intention to become a prosumer. An overall scale measuring “acceptance of a renewable energy solution” was used as the dependent variable, allowing us to quantify the five factors that contribute to measuring: awareness of environmental issues and climate change; environmental attitudes; social influence; and environmental risk perception. Three variables can significantly measure and predict the scores of the “Acceptance in becoming a prosumer” ad hoc scale. Variable 1. Attitude: the agreement to specific environmental issues and global climate change issues of concerns and evaluations towards a behavioural intention. Variable 2. Economic incentive: the perceived behavioural control and its related environmental risk perception, in terms of perceived short-term benefits and long-term costs, both part of the decision-making process as expected outcomes of the behaviour itself. Variable 3. Age: despite fewer economic possibilities, younger adults seem to be more sensitive to environmental dimensions and issues as opposed to older adults. This research can facilitate policymakers and relevant stakeholders to better understand which relevant psycho-sociological factors are intervening in these processes and what and how specifically target when proposing change towards sustainable energy production and consumption.

Keywords: behavioural intention, environmental risk perception, prosumer, renewable energy technology, social acceptance

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Authors: Leila Noori, Rosario Barone, Francesca Rappa, Antonella Marino Gammazza, Alessandra Maria Vitale, Giuseppe Donato Mangano, Giusy Sentiero, Filippo Macaluso, Kathryn H. Myburgh, Francesco Cappello, Federica Scalia

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The neuromuscular system controls, directs, and allows movement of the body through the action of neural circuits, which include motor neurons, sensory neurons, and skeletal muscle fibers. Protein homeostasis of the involved cytotypes appears crucial to maintain the correct and prolonged functions of the neuromuscular system, and both neuronal cells and skeletal muscle fibers express significant quantities of protein chaperones, the molecular machinery responsible to maintain the protein turnover. Genetic mutations or defective post-translational modifications of molecular chaperones (i.e., genetic or acquired chaperonopathies) may lead to neuromuscular disorders called as neurochaperonopathies. The limited knowledge of the effects of the defective chaperones on skeletal muscle fibers and neurons impedes the progression of therapeutic approaches. A distinct genetic variation of CCT5 gene encoding for the subunit 5 of the chaperonin CCT (Chaperonin Containing TCP1; also known as TRiC, TCP1 Ring Complex) was recently described associated with severe distal motor neuropathy by our team. In this study, we investigated the histopathological abnormalities of the skeletal muscle biopsy of the pediatric patient affected by the mutation Leu224Val in the CCT5 subunit. We provide molecular and structural features of the diseased skeletal muscle tissue that we believe may be useful to identify undiagnosed cases of this rare genetic disorder. We investigated the histological abnormalities of the affected tissue via hematoxylin and eosin staining. Then we used immunofluorescence and qPCR techniques to explore the expression and distribution of CCT5 in diseased and healthy skeletal muscle tissue. Immunofluorescence and immunohistochemistry assays were performed to study the sarcomeric and structural proteins of skeletal muscle, including actin, myosin, tubulin, troponin-T, telethonin, and titin. We performed Western blot to examine the protein expression of CCT5 and some heat shock proteins, Hsp90, Hsp60, Hsp27, and α-B crystallin, along with the main client proteins of the CCT5, actin, and tubulin. Our findings revealed muscular atrophy, abnormal morphology, and different sizes of muscle fibers in affected tissue. The swollen nuclei and wide interfiber spaces were seen. Expression of CCT5 had been decreased and showed a different distribution pattern in the affected tissue. Altered expression, distribution, and bandage pattern were detected by confocal microscopy for the interested muscular proteins in tissue from the patient compared to the healthy control. Protein levels of the studied Hsps normally located at the Z-disk were reduced. Western blot results showed increased levels of the actin and tubulin proteins in the diseased skeletal muscle biopsy compared to healthy tissue. Chaperones must be expressed at high levels in skeletal muscle to counteract various stressors such as mechanical, oxidative, and thermal crises; therefore, it seems relevant that defects of molecular chaperones may result in damaged skeletal muscle fibers. So far, several chaperones or cochaperones involved in neuromuscular disorders have been defined. Our study shows that alteration of the CCT5 subunit is associated with the damaged structure of skeletal muscle fibers and alterations of chaperone system components and paves the way to explore possible alternative substrates of chaperonin CCT. However, further studies are underway to investigate the CCT mechanisms of action to design applicable therapeutic strategies.

Keywords: molecular chaperones, neurochaperonopathy, neuromuscular system, protein homeostasis

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