Inherited retinal dystrophies are often associated with mutations in the genes involved in the phototransduction cascade in photoreceptors, a paradigmatic signaling pathway mediated by G protein-coupled receptors. Photoreceptor viability is strictly dependent on the levels of the second messengers cGMP and Ca²⁺. Here we explored the possibility of modulating the phototransduction cascade in mouse rods using direct or liposome-mediated administration of a recombinant protein crucial for regulating the interplay of the second messengers in photoreceptor outer segments. The effects of administration of the free and liposome-encapsulated human guanylate cyclase-activating protein 1 (GCAP1) were compared in biological systems of increasing complexity (in cyto, ex vivo, and in vivo). The analysis of protein biodistribution and the direct measurement of functional alteration in rod photoresponses show that the exogenous GCAP1 protein is fully incorporated into the mouse retina and photoreceptor outer segments. Furthermore, only in the presence of a point mutation associated with cone-rod dystrophy in humans p.(E111V), protein delivery induces a disease-like electrophysiological phenotype, consistent with constitutive activation of the retinal guanylate cyclase. Our study demonstrates that both direct and liposome-mediated protein delivery are powerful complementary tools for targeting signaling cascades in neuronal cells, which could be particularly important for the treatment of autosomal dominant genetic diseases. Supplementary Information The online version contains supplementary material available at 10.1007/s00018-023-05022-0.
Digital anthropometry by three-dimensional optical imaging systems and smartphones has recently been shown to provide non-invasive, precise, and accurate anthropometric and body composition measurements. To our knowledge, no previous study performed smartphone-based digital anthropometric assessments in young athletes. The aim of this study was to investigate the reproducibly and validity of smartphone-based estimation of anthropometric and body composition parameters in youth soccer players. A convenience sample of 124 male players and 69 female players (median ages of 16.2 and 15.5 years, respectively) was recruited. Measurements of body weight and height, one whole-body Dual-Energy X-ray Absorptiometry (DXA) scan, and acquisition of optical images (performed in duplicate by the Mobile Fit app to obtain two avatars for each player) were performed. The reproducibility analysis showed percent standard error of measurement values < 10% for all anthropometric and body composition measurements, thus indicating high agreement between the measurements obtained for the two avatars. Mobile Fit app overestimated the body fat percentage with respect to DXA (average overestimation of + 3.7% in males and + 4.6% in females), while it underestimated the total lean mass (− 2.6 kg in males and − 2.5 kg in females) and the appendicular lean mass (− 10.5 kg in males and − 5.5 kg in females). Using data of the soccer players, we reparameterized the equations previously proposed to estimate the body fat percentage and the appendicular lean mass and we obtained new equations that can be used in youth athletes for body composition assessment through conventional anthropometrics-based prediction models.
Background Pancreatic ductal adenocarcinoma (PDAC) is a lethal disease. This is due to its aggressive course, late diagnosis and its intrinsic drugs resistance. The complexity of the tumor, in terms of cell components and heterogeneity, has led to the approval of few therapies with limited efficacy. The study of the early stages of carcinogenesis provides the opportunity for the identification of actionable pathways that underpin therapeutic resistance. Methods We analyzed 43 Intraductal papillary mucinous neoplasms (IPMN) (12 Low-grade and 31 High-grade) by Spatial Transcriptomics. Mouse and human pancreatic cancer organoids and T cells interaction platforms were established to test the role of mucins expression on T cells activity. Syngeneic mouse model of PDAC was used to explore the impact of mucins downregulation on standard therapy efficacy. Results Spatial transcriptomics showed that mucin O -glycosylation pathway is increased in the progression from low-grade to high-grade IPMN. We identified GCNT3, a master regulator of mucins expression, as an actionable target of this pathway by talniflumate. We showed that talniflumate impaired mucins expression increasing T cell activation and recognition using both mouse and human organoid interaction platforms. In vivo experiments showed that talniflumate was able to increase the efficacy of the chemotherapy by boosting immune infiltration. Conclusions Finally, we demonstrated that combination of talniflumate, an anti-inflammatory drug, with chemotherapy effectively improves anti-tumor effect in PDAC.
In plants, glutamate dehydrogenase (GDH) is an ubiquitous enzyme that catalyzes the reversible amination of 2‐oxoglutarate in glutamate. It contributes to both the amino acid homeostasis and the management of intracellular ammonium, and it is regarded as a key player at the junction of carbon and nitrogen assimilation pathways. To date, information about the GDH of terrestrial plants refers to a very few species only. We focused on selected species belonging to the division Marchantiophyta, providing the first panoramic overview of biochemical and functional features of GDH in liverworts. Native electrophoretic analyses showed an isoenzymatic profile less complex than what was reported for Arabidposis thaliana and other angiosperms: the presence of a single isoform corresponding to an α‐homohexamer, differently prone to thermal inactivation on a species‐ and organ‐basis, was found. Sequence analysis conducted on amino acid sequences confirmed a high similarity of GDH in modern liverworts with the GDH2 protein of A. thaliana , strengthening the hypothesis that the duplication event that gave origin to GDH1 ‐homolog gene from GDH2 occurred after the evolutionary bifurcation that separated bryophytes and tracheophytes. Experiments conducted on Marchantia polymorpha and Calypogeia fissa grown in vitro and compared to A. thaliana demonstrated through in gel activity detection and monodimensional Western Blot that the aminating activity of GDH resulted in strongly enhanced responses to ammonium excess in liverworts as well, even if at a different extent compared to Arabidopsis and other vascular species. The comparative analysis by bi‐dimensional Western Blot suggested that the regulation of the enzyme could be, at least partially, untied from the protein post‐translational pattern. Finally, immuno‐electron microscopy revealed that the GDH enzyme localizes at the subcellular level in both mitochondria and chloroplasts of parenchyma and is specifically associated to the endomembrane system in liverworts.
This is a comparative study of two countries’ legal and policy actions to stem the spread of COVID-19 in the workplace, focusing on Italy and the United States (US). Both nations sustained great loss of life and high infection rates during the first years of the pandemic. This chapter examines how Italy and the US approached COVID-19 vaccine mandates for workers. Of particular interest are the regulatory choices made, including the choice not to regulate, and the consequences of those choices on the employment relationship. Additionally revealing are the legal grounds upon which regulatory actions were challenged, and how courts balanced the interests at stake. Finally, the way in which the debates over workplace vaccine mandates were framed illuminate national culture and the extent to which each country views labor rights as human rights. To provide context for these insights, this chapter examines convergence and divergence in the two countries’ initial responses to the global health emergency presented by COVID-19, and the way in which workplace vaccine mandates were initially embraced.
Importance High-sensitivity troponin tests can detect even milder cardiac troponin elevations in plasma, beyond the threshold of conventional troponin tests. Whether detection of low-grade cardiac troponin elevation is associated with outcomes of patients with hemodynamically stable pulmonary embolism (PE) and helps with risk stratification is unknown. Objective To determine the association between high-sensitivity cardiac troponin I (hs-cTnI) compared with conventional cardiac troponin I (cTnI) and PE risk designations according to the European Society of Cardiology (ESC) 2019 classification scheme and clinical outcomes in patients with hemodynamically stable PE. Design, Setting, and Participants This is a post hoc analysis of data from the prospective Prognostic Value of Computed Tomography (PROTECT) multicenter cohort study enrolling patients from 12 hospital emergency departments in Spain. In this analysis, cTnI and hs-cTnI were compared with respect to ESC risk designation, and the association between troponin values and a complicated course after PE diagnosis was evaluated. Of 848 patients enrolled in PROTECT, 834 (98.3%) had hsTnI and cTnI values available and were included in the present analysis. Data were analyzed from May to December 2022. Exposures Troponin blood testing with cTnI (threshold of >0.05 ng/mL) vs hs-cTnI (threshold of >0.029 ng/mL) assays at the time of PE diagnosis. Main Outcomes Complicated course, defined as hemodynamic collapse, recurrent PE, or all-cause death, within 30 days after PE. Results Of 834 patients (mean [SEM] age, 67.5 [0.6] years; 424 [50.8%] female), 139 (16.7%) had elevated cTnI and 264 (31.7%) elevated hs-TnI, respectively. During follow-up, 62 patients (7.4%; 95% CI, 5.7-9.4) had a complicated course. Analyzing troponin elevation as a binary variable, elevated cTnI (odds ratio [OR], 2.84; 95% CI, 1.62-4.98) but not hs-cTnI (OR, 1.12; 95% CI, 0.65-1.93) was associated with increased odds of a complicated course. Of 125 patients who had elevated hs-cTnI but normal cTnI, none (0; 95% CI, 0.0-2.9) developed a complicated course. Using the 2019 ESC risk stratification scheme, hs-TnI classified fewer patients as low risk compared with cTnI. Among 78 patients designated as ESC low risk when using cTnI but not with hsTnI, none (0; 95% CI, 0.0-4.6) had a complicated course. Conclusions and Relevance In this study of patients with hemodynamically stable PE, hs-cTnI identified modest elevations in cardiac troponin levels. However, the results did not provide additive clinical value compared with cTnI. These findings suggest that use of hs-cTnI may result in overestimation of the risk in patients with stable PE.
Radioligand therapy (RLT) with lutetium (177Lu) oxodotreotide is an approved therapy in combination with somatostatin analogues (SSAs) for patients with advanced, well-differentiated G1–G2, gastro-entero-pancreatic neuroendocrine tumours (GEP-NETs) that progress on SSAs. We conducted a series of round table meetings throughout Italy to identify issues related to RLT delivery to patients with GEP-NETs. Four key issues were identified: (1) the proper definition of tumour progression prior to RLT initiation; (2) the impact of RLT in patients with bone metastases and/or high hepatic tumour burden; (3) the optimal follow-up protocol after RLT; and (4) organisational issues related to RLT use and managerial implications. This article reviews the literature relating to the aforementioned issues and makes recommendations based on available evidence and Italian NET experts’ opinions. In particular, the group recommends the development of a diagnostic–therapeutic care pathway (DTCP) for patients undergoing RLT which provides systematic guidance but can still be individualised for each patient’s clinical and psychosocial needs. A DTCP may clarify the diagnostic, therapeutic and post-treatment monitoring process, and improve communication and the coordination of care between hub and spoke centres. The DTCP may also contribute to changes in the care process related to the 2013/59/EURATOM Directive and to the definition of costs when planning for future or updated reimbursement of RLT in Italy.
One of the main technological barriers hindering the development of active industrial exoskeleton is today represented by the lack of suitable payload estimation algorithms characterized by high accuracy and low calibration time. The knowledge of the payload enables exoskeletons to dynamically provide the required assistance to the user. This work proposes a payload estimation methodology based on personalized Electromyography-driven musculoskeletal models (pEMS) combined with a payload estimation method we called "delta torque" that allows the decoupling of payload dynamical properties from human dynamical properties. The contribution of this work lies in the conceptualization of such methodology and its validation considering human operators during industrial lifting tasks. With respect to existing solutions often based on machine learning, our methodology requires smaller training datasets and can better generalize across different payloads and tasks. The proposed payload estimation methodology has been validated on lifting tasks with 0kg, 5kg, 10kg and 15kg, resulting in an average MAE of about 1.4 Kg. Even if 5kg and 10Kg lifting tasks were out of the training set, the MAE related to these tasks are 1.6 kg and 1.1 kg, respectively, demonstrating the generalizing property of the proposed methodology. To the best of the authors’ knowledge, this is the first time that an EMG-driven model-based approach is proposed for human payload estimation.
Crowdfunding campaigns have recently promoted a range of new business models in different contexts. This study investigates crowdfunding in the accommodation realm from a socio-cultural perspective and across its international dynamics. Drawing on complexity theory, the study explores the successful case of CleanBnb, the leading crowdfunded company in the Italian short-term rental market, and informs hospitality actors on the coping strategies implemented to challenge the Covid-19 pandemic. The study adopts a case study approach, combining primary data collected through an in-depth interview of the CEO and the analysis of secondary data from different company reports. The results highlight the importance of (1) business diversification, (2) grouping opportunities and (3) widening of service range as key factors in pandemic business survival for start-ups operating in the accommodation realm. The study finally discusses post-pandemic scenarios for both the traditional hotel industry and sharing economy operators by offering managerial insights.
Purpose Breast cancer (BC) is the most common malignancy that affects women, and it is, to date, their leading cause of death. Luminal A molecular subtype accounts for 40% of BC and is characterized by hormone receptors positive/human epidermal growth factor 2 expression and current treatment consists of surgery plus aromatase inhibitor therapy. Interestingly, several studies demonstrated that the heavy metal cadmium (Cd), classified as a group 1 human carcinogen and widely spread in the environment, exerts estrogen-like activities in several tissues and suggested an intriguing relationship between increased Cd exposure and BC incidence. Thus, aim of this study was to evaluate effects of Cd on Luminal A BC estrogen receptor (ER) positive/progesterone receptor positive cell models in vitro to characterize the mechanism(s) involved in breast cell homeostasis disruption. Methods T47D and MCF7 were exposed to Cd (0.5–1 µM) for 6–24 h to evaluate potential alterations in: cells viability, steroid receptors and intracellular signaling by western blot. Moreover, we evaluated the expression of inflammatory cytokines interleukin by RT-PCR. Results Our results showed a significant induction of androgen receptor (AR) and an increased AR/ER ratio. Further, Cd exposure increased pro-inflammatory cytokines interleukin (IL)6, IL8 and tumor necrosis factor α levels. Finally, as previously demonstrated by our group, Cd alters pathways such as mitogen-activated protein kinase family and protein kinase B. Conclusion In conclusion, our study demonstrates that Cd modifies the expression and pattern of ERs and AR in BC cell lines, suggesting an alteration of BC cells homeostasis, likely predisposing to a carcinogenetic microenvironment.
Background Surgery with negative resection margins and appropriate lymphadenectomy is the only curative treatment for gastric cancer. The adequate proximal resection margins length in European guidelines is between 5 to 8 centimeters depending on the histological subtype. However, these recommendations are based on outdated studies with limited data. The peri-operative treatment modalities have evolved, and the application of these resection margin cut-offs may not be as uniform as before. The aim of our study was to assess the current European practice in gastric cancer surgery with particular attention to resection margins. Methods A panel of 13 international experts composed of surgeons, pathologists, and oncologists developed a web-based survey containing 45 questions about center demographic, resection margin strategy, and post-operative care. The survey was distributed to Upper Gastrointestinal (UGI) surgeons through UGI and visceral surgical societies mailing lists and social media between February and June 2022. Participants were excluded if they did not answer a minimum of 80% completion of the questionary). Results 172 surgeons from 154 centers in 19 countries responded to the survey. The most represented countries were Germany 60 (35%), Switzerland 30 (18%) and Italy 16 (9%). The mean surgical years’ experience of the responders was 20.8 and their caseload was 23.2 cases per year. For intestinal type gastric tumor, a proximal resection margin length of a minimum of 5 centimeters was advocated by 119 (77%) of the responders. For diffuse, a resection margin of a minimum of 8 cm was applied by 98 (63%) of the surgeons. Macroscopic inspection of the specimen was routinely performed by 110 respondents (64%). Conclusions The real-world data from our questionnaire shows that the length of resection margins proposed by actual guidelines are not strictly applied in European centers. Further data from clinical research are needed to standardize resection margin cut-off. This could help European surgeons to propose more organ-sparing strategies to their patients.
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