Fatty Hepatocytes Induce Skeletal Muscle Atrophy In Vitro: A New 3D Platform to Study the Protective Effect of Albumin in Non-Alcoholic Fatty Liver

The liver neutralizes endogenous and exogenous toxins and metabolites, being metabolically interconnected with many organs. Numerous clinical and experimental studies show a strong association between Non-alcoholic fatty liver disease (NAFLD) and loss of skeletal muscle mass known as sarcopenia. Liver transplantation solves the hepatic-related insufficiencies, but it is unable to revert sarcopenia. Knowing the mechanism(s) by which different organs communicate with each other is crucial to improve the drug development that still relies on the two-dimensional models. However, those models fail to mimic the pathological features of the disease. Here, both liver and skeletal muscle cells were encapsulated in gelatin methacryloyl and carboxymethylcellulose to recreate the disease’s phenotype in vitro. The 3D hepatocytes were challenged with non-esterified fatty acids (NEFAs) inducing features of Non-alcoholic fatty liver (NAFL) such as lipid accumulation, metabolic activity impairment and apoptosis. The 3D skeletal muscle tissues incubated with supernatant from fatty hepatocytes displayed loss of maturation and atrophy. This study demonstrates the connection between the liver and the skeletal muscle in NAFL, narrowing down the players for potential treatments. The tool herein presented was employed as a customizable 3D in vitro platform to assess the protective effect of albumin on both hepatocytes and myotubes

Healthcare Service Quality Evaluation in a Community-Oriented Primary Care Center, Italy

Community-oriented primary care (COPC) is an inclusive healthcare approach that combines individual care with a population-based outlook, striving to offer effective and equitable services. This study concentrates on assessing the perceived quality of a "Casa della Comunita" (CdC) implemented by the Romagna Local Health Authority, which embraces the COPC model. Through the examination of user experiences, the study aims to comprehend the influence of the CdC's care delivery model on the community's perception of service quality. From 13-18 March 2023, paper questionnaires were distributed by trained healthcare professionals and volunteers. The cross-sectional study enrolled participants aged 18 or older, capable of understanding written Italian, and willing to take part voluntarily. A total of 741 questionnaires were collected, resulting in an overall acceptance rate of 85.6%. Among the respondents, 37.9% were female, with an average age of 55.4 & PLUSMN; 16.2 years. While the respondents generally held a positive view of the quality, the results displayed varying levels of satisfaction across the different areas. Multivariate analysis revealed significant associations between factors such as gender, employment status, financial resources, education level, and distance from the healthcare center with the perceived quality of the facility in terms of accessibility, environment, staff, continuity of care, and overall satisfaction. The study yielded valuable insights, identifying strengths and areas for improvement and underscoring the importance of ongoing monitoring studies to enhance patient satisfaction continuously

Use of Medications for Management of Alzheimer’s Disease in Ontario’s Home Care Population

Abstract Background: Home care is an important care setting for those with Alzheimer’s disease (AD). It provides support that allows individuals with AD to remain at home and may delay the transition to long-term care homes. Many clients with AD receive medications that are used for managing the symptoms of AD: cholinesterase inhibitors (ChEIs) and memantine. Ontario’s provincial drug benefit plan (ODB) provides subsidies for some of these medications based on specific clinical criteria. These AD medications are costly and can have significant side effects, so it is important to understand how they are being used in practice. Objectives: The objectives of this study were to report the proportion taking AD medications and which types were taken, show the change in receipt of AD medications over time, and show the covariates that were independently associated with receiving AD medications. Methods: Analysis of secondary data was performed on the provincial home care dataset. All home care clients receiving long-term home care services were assessed using the RAI-Home Care (RAI-HC), which is a comprehensive and standardized assessment. One assessment from each individual over the age of 65 who was assessed between January 2004 and September 2008 was used, for a final sample size of 321,013. Results: Overall, 65% of clients with a diagnosis of AD were receiving an AD medication. Logistic regression analysis among those diagnosed with AD showed that increased physical impairment and clinical complexity were associated with decreased odds of receiving AD medication. Contraindicating diagnoses such as congestive heart failure, lack of medical oversight and needing to make economic tradeoffs were also associated with decreased odds of receiving AD medication. Conclusions: The multivariate model showed trends of rational prescribing, such as clients with contraindicating diagnoses or very high clinical complexity having decreased odds of receiving AD medications. At the same time, evidence of structural barriers to receiving the medications was shown. There is debate about the cost-effectiveness of these medications. The provincial government could consider expanding ODB guidelines to include all AD medications for those with all levels of cognitive impairment, but further analyses involving longitudinal outcomes available in this dataset should be performed to ensure it would be in the public interest

How future surgery will benefit from SARS-COV-2-related measures: a SPIGC survey conveying the perspective of Italian surgeons

COVID-19 negatively affected surgical activity, but the potential benefits resulting from adopted measures remain unclear. The aim of this study was to evaluate the change in surgical activity and potential benefit from COVID-19 measures in perspective of Italian surgeons on behalf of SPIGC. A nationwide online survey on surgical practice before, during, and after COVID-19 pandemic was conducted in March-April 2022 (NCT:05323851). Effects of COVID-19 hospital-related measures on surgical patients' management and personal professional development across surgical specialties were explored. Data on demographics, pre-operative/peri-operative/post-operative management, and professional development were collected. Outcomes were matched with the corresponding volume. Four hundred and seventy-three respondents were included in final analysis across 14 surgical specialties. Since SARS-CoV-2 pandemic, application of telematic consultations (4.1% vs. 21.6%; p < 0.0001) and diagnostic evaluations (16.4% vs. 42.2%; p < 0.0001) increased. Elective surgical activities significantly reduced and surgeons opted more frequently for conservative management with a possible indication for elective (26.3% vs. 35.7%; p < 0.0001) or urgent (20.4% vs. 38.5%; p < 0.0001) surgery. All new COVID-related measures are perceived to be maintained in the future. Surgeons' personal education online increased from 12.6% (pre-COVID) to 86.6% (post-COVID; p < 0.0001). Online educational activities are considered a beneficial effect from COVID pandemic (56.4%). COVID-19 had a great impact on surgical specialties, with significant reduction of operation volume. However, some forced changes turned out to be benefits. Isolation measures pushed the use of telemedicine and telemetric devices for outpatient practice and favored communication for educational purposes and surgeon-patient/family communication. From the Italian surgeons' perspective, COVID-related measures will continue to influence future surgical clinical practice

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

The Key Role of IP6K: A Novel Target for Anticancer Treatments?

Inositol and its phosphate metabolites play a pivotal role in several biochemical pathways and gene expression regulation: inositol pyrophosphates (PP-IPs) have been increasingly appreciated as key signaling modulators. Fluctuations in their intracellular levels hugely impact the transfer of phosphates and the phosphorylation status of several target proteins. Pharmacological modulation of the proteins associated with PP-IP activities has proved to be beneficial in various pathological settings. IP 7 has been extensively studied and found to play a key role in pathways associated with PP-IP activities. Three inositol hexakisphosphate kinase (IP 6 K) isoforms regulate IP 7 synthesis in mammals. Genomic deletion or enzymic inhibition of IP 6 K1 has been shown to reduce cell invasiveness and migration capacity, protecting against chemical-induced carcinogenesis. IP 6 K1 could therefore be a useful target in anticancer treatment. Here, we summarize the current understanding that established IP 6 K1 and the other IP 6 K isoforms as possible targets for cancer therapy. However, it will be necessary to determine whether pharmacological inhibition of IP 6 K is safe enough to begin clinical study. The development of safe and selective inhibitors of IP 6 K isoforms is required to minimize undesirable effects

The activation of miR-125a-5p/IP6K1 axis in breast cancer cells upon treatment with myo-Inositol.

Several studies have been performed with the aim of identifying drugs able in inhibiting Epithelial-Mesenchymal Transition (EMT), chiefly by blocking PI3K/Akt pathway. We have already demonstrated that treatment with myo-Inositol at the pharmacological dose can block EMT in breast cancer cells by downregulating PI3K/Akt and inducing changes in cytoskeletal architecture. Herewith, we investigated the mechanism of action of myo-inositol in both highly (MDA-MB-231) and low (MCF-7) invasive human breast cancer cells. After 30’ and 24h from treatment, gene expression analysis revealed a significant downregulation of Pi3k and Psen1 after 30’ in both cell lines. Psen1 downregulation was maintained in MDA-MB-231 at 24h. Likewise, we explored the modulation of Ip6k1, Dnmt3b, Isyna1 and p53. In MDA-MB-231, a strong downregulation of Ip6k1 expression was recorded at 30’ and 24h, whilst Dnmt3b was reduced only at 30’. On the contrary, in MCF-7, Ip6k1 downregulation was unexpectedly associated to the upregulation of Dnmt3b at 30’. IP6K1 is a key enzyme of inositol metabolism, inhibits ISYNA1, probably inducing de novo DNA methylation (i.e., DNMT3B). Furthermore, IP6K1 inhibition correlates with a decrease of cancer cells motility. The upregulation of Isyna1 was observed in both cell lines at 30’, together with p53. ISYNA1 activates myo-Inositol intracellular biosynthesis starting from glucose-6-phosphate. In this activation, p53 plays a key role in binding Isyna1 promoter and eventually enabling its expression. Western-blot of MDA-MB-231 confirmed that changes in gene expression were also mirrored by concurrently modifications in IP6K1 and p53 protein levels, altogether with a decrease of both MDM2 and YAP/TAZ. It is worth noting that in MCF-7, no changes were observed in protein levels. In-silico analysis was performed using TCGA miRNA-Seq data to identify differentially expressed miRNAs between normal and tumoral tissue in breast cancer patients. To further gain mechanistic insights on myo-Inositol effects, we compared these data with main differentially expressed cancer-related miRNAs in MDA-MB-231 cells after 30’ from treatment. This analysis allowed to identify two mRNAs, downregulated in tumor tissues, that were significantly increased with myo-Inositol: miR-92a-3p and miR-125a-5p. Using DIANA tools, miR-92a-3p was predicted to interact with Notch-1 and PI3K, linking it to cytoskeletal rearrangement. Moreover, a strong interaction was predicted between miR-125a-5p and IP6K1 in 3’-UTR site. Indeed, the upregulation of miR-125a-5p is usually correlated with metastasis inhibition in breast cancer. In MDA-MB-231, miR-125a-5p upregulation was maintained at 24h, while in MCF-7 was slightly upregulated at 30’ and downregulated at 24h. Our results suggest that myo-Inositol causes early changes in gene expression, probably led by miRNAs and methylation remodeling. Elucidation of the role of miR-125a-5p/IP6K1 axis will reveal strategies for molecular targeted therapies in breast cancer

Myo-Inositol treatment inhibits motility in triple negative breast cancer via miR-125a-5p/IP6K1 axis

Background: Several researches have been performed with the aim of identifying drugs able in blocking PI3K/Akt pathway. We have already demonstrated that myo-Inositol (myo-Ins) treatment can block EMT in breast cancer cells by downregulating PI3K/Akt and inducing changes in cytoskeletal architecture. Aim: Herein, we set our experiments to investigate migration/invasiveness inhibition though in vitro and in vivo models upon myo-Ins administration. Methods: In vitro experiments were performed using both mesenchymal-like (MDA-MB-231) and epithelial-like (MCF-7) invasive human breast cancer cells. We used transwell assays for in vitro and Zebrafish embryos as in vivo models to evaluate migration and invasiveness. The expression of key genes involved in the mechanism was evaluated by qPCR, while gain- and loss-of-function approaches allowed identifying the specific dynamical relationships. Results: Myo-Ins inhibits motility and invasiveness only in MDA-MB-231 cells both in vitro and in vivo. In MDA cells, miR-125a-5p upregulation was linked to IP6K1 downregulation triggered by myo-Ins treatment. Silencing and overexpression experiments confirmed the key role of miR-125a-5p/IP6K1 axis in blocking cell motility. This effect was demonstrated to be myo-Ins-dependent MDM2 inhibition. Given that MDM2 in MCF-7 cells was unaffected by treatment, in these cells myo-Ins was unable in antagonizing motility. Conversely, both miR-125a-5p and IP6K1 were not modulated. However, MDM2 silencing restore sensitivity to myo-Ins, thus leading to a significant inhibition of the MCF-7 cells motility capability. Conclusions: Our results suggest that myo-Ins can inhibit motility in triple negative breast cancer. Such an effect is likely mediated by MDM2 inhibition, which, in turn, triggers a complex tumor reversion promoted by the miR-125a-5p/IP6K1 axis modulation. Elucidation of the role of miR-125a- 5p/IP6K1 axis will reveal strategies for molecular targeted therapies in breast cancer

Risk stratifying asymptomatic left ventricular systolic dysfunction in the community. beyond left ventricular ejection fraction

Aims: Midwall fractional shortening (MWFS) is a measure of left ventricular (LV) systolic function that is more reliable in case of concentric LV geometry compared to LV ejection fraction (LVEF). We hypothesized that MWFS might predict heart failure (HF) and death in a high-risk asymptomatic population, beyond other echocardiographic parameters. Methods and results: Among 4047 subjects aged ≥55/≤80 years followed by 10 general practitioners in northern Italy, the DAVID-Berg study prospectively enrolled 623 asymptomatic outpatients at increased risk for HF. Baseline evaluation included clinical visit, electrocardiogram, N-terminal pro-brain natriuretic peptide (NT-proBNP), and echocardiogram. Mean age of the population was 69 ± 7 years, 56% were men, 88% had hypertension, mean LVEF was 61 ± 9%, and mean MWFS 16.2 ± 3.3. During a median follow-up of 5.7 years, 95 subjects experienced HF/death events. At Cox analysis, lower MWFS was the only echocardiographic parameter, among structural/functional ones, associated with higher risk of HF/death [hazard ratio (HR) 0.89, 95% confidence interval (CI) 0.84-0.95, Padjusted < 0.001]. The risk of HF/death related to clinical data and NT-proBNP (baseline model) was reclassified by echocardiography only when MWFS was included into the model (baseline C-statistics 0.761; adding conventional structural/functional echocardiographic data 0.776, P = 0.09; adding MWFS 0.791, P = 0.007). Compared to subjects with normal LVEF and MWFS, only subjects with combined systolic dysfunction (11% of the population) were at higher risk (P = 0.001 for both abnormal; P > 0.24 for either LVEF or MWFS abnormal). Conclusion: DAVID-Berg data suggest to include MWFS assessment in clinical practice, a simple and reliable echocardiographic parameter able to improve risk stratification in subjects at high risk for HF