Therapeutic Intervention Research Articles

Luminescent Alendronic Acid-Conjugated Micellar Nanostructures for Potential Application in the Bone-Targeted Delivery of Cholecalciferol

Vitamin D, an essential micronutrient crucial for skeletal integrity and various non-skeletal physiological functions, exhibits limited bioavailability and stability in vivo. This study is focused on the development of polyethylene glycol (PEG)-grafted phospholipid micellar nanostructures co-encapsulating vitamin D3 and conjugated with alendronic acid, aimed at active bone targeting. Furthermore, these nanostructures are rendered optically traceable in the UV–visible region of the electromagnetic spectrum via the simultaneous encapsulation of vitamin D3 with carbon dots, a newly emerging class of fluorescents, biocompatible nanoparticles characterized by their resistance to photobleaching and environmental friendliness, which hold promise for future in vitro bioimaging studies. A systematic investigation is conducted to optimize experimental parameters for the preparation of micellar nanostructures with an average hydrodynamic diameter below 200 nm, ensuring colloidal stability in physiological media while preserving the optical luminescent properties of the encapsulated carbon dots. Comprehensive chemical-physical characterization of these micellar nanostructures is performed employing optical and morphological techniques. Furthermore, their binding affinity for the principal inorganic constituent of bone tissue is assessed through a binding assay with hydroxyapatite nanoparticles, indicating significant potential for active bone-targeting. These formulated nanostructures hold promise for novel therapeutic interventions to address skeletal-related complications in cancer affected patients in the future.

Abstract IA008: Dissecting innate immune mechanisms of resistance to immune checkpoint blockade therapy in bladder cancer

Abstract We have explored the TME of bladder cancer (BC) leveraging single-cell RNA sequencing (scRNAseq) and clinical trial data to ascertain the complexity of tumor-infiltrated immune ecllls in BC. As a first step we identify distinct tumor-associated macrophage (TAM) populations associated with immune checkpoint blockade (ICB) response and resistance. Through plasma cytokine profiling, in vitro assays, and spatial analyses, we uncover novel cytokine-driven axes influencing these TAM populations and their impact on T cells, offering insights into enhancing anti-tumor T cell immunity in BC. Second, we investigate the potential of urine-derived cells (UDCs) as surrogates for monitoring the BC TME. Our scRNAseq atlas of BC patient UDCs reveals a diverse composition of immune cell populations, strikingly transcriptionally similar to tumor immune cells and containing immune subsets absent or rare in peripheral blood, suggesting their utility as biomarkers for response to therapeutic intervention. Collectively, these findings provide new insights into the immune landscape of BC and provide putative targets to enhance existing immunotherapy strategies in BC. Citation Format: Nina Bhardwaj. Dissecting innate immune mechanisms of resistance to immune checkpoint blockade therapy in bladder cancer [abstract]. In: Proceedings of the AACR Special Conference on Bladder Cancer: Transforming the Field; 2024 May 17-20; Charlotte, NC. Philadelphia (PA): AACR; Clin Cancer Res 2024;30(10_Suppl):Abstract nr IA008.

Limning of HIF-2 and HIF-3 in the Tumor Microenvironment: Developing Concepts for the Treatment of Hypoxic Cancer.

Hypoxia, characterized by insufficient oxygen supply to tissues, is a significant factor in tumor growth and resistance to treatment. The hypoxia-inducible factor (HIF) signaling pathway is activated when oxygen levels decline, influencing cell activities and promoting tumor progression. HIF-1α and HIF-2α are the main targets for therapeutic intervention in tumors. Nevertheless, the significance of HIF-2α is often overlooked. This review examines the physiological role of HIF-2α in tumor growth and its involvement in tumor growth. HIFs, composed of hypoxia-responsive α and oxygeninsensitive β subunits, play a crucial role in controlling gene expression in both normal and solid tumor tissues under low oxygen levels. HIF-3α, formerly considered a detrimental modulator of HIF-regulated genes, exerts a transcriptional regulatory role by inhibiting gene expression through competition with HIF-1α and HIF-2α for binding to transcriptional sites in target genes under hypoxia. Recent research indicates that various HIF-3 variants exhibit distinct and potentially contrasting functionalities. Hypoxia often occurs during the initiation and progression of cancer formation. Recent research has discovered that HIF-2α, also known as endothelial PAS domain protein 1, has a significant impact on tumors. HIF-2α is a significant cancer-causing gene and a crucial predictor of prognosis in non-small cell lung cancer. However, due to limited research investigating the relationship between HIF-2α and small-cell lung cancer, it is not possible to reach a definitive conclusion. HIF-2α plays a vital function in cancer by preserving the stemness of cancer cells. This review provides a comprehensive overview of HIF-2 and the role of HIF-3 in various cancer-related processes, as well as its potential as a targeted therapeutic approach.

Unraveling the path to osteoarthritis management: targeting chondrocyte apoptosis for therapeutic intervention

Osteoarthritis (OA) is a chronic disease affecting joints and further causing disabilities. This disease affects around 240 million people worldwide. It is a multifactorial disease, and its etiology is difficult to determine. Although numerous therapeutic strategies are available, the therapies are aimed at reducing pain and improving patients’ quality of life. Hence, there is an urgent need to develop disease-modifying drugs (DMOAD) that can reverse or halt OA progression. Apoptosis is a cell removal process that is important in maintaining homeostatic mechanisms in the development and sustaining cell population. The apoptosis of chondrocytes is believed to play an important role in OA progression due to poor chondrocytes self-repair abilities to maintain the extracellular matrix (ECM). Hence, targeting chondrocyte apoptosis can be one of the potential therapeutic strategies in OA management. There are various mediators and targets available to inhibit apoptosis such as autophagy, endoplasmic reticulum (ER) stress, oxidative stress, and inflammation. As such, this review highlights the importance and potential targets that can be aimed to reduce chondrocyte apoptosis.

Greater mechanistic understanding of the cutaneous pathogenesis of Stevens-Johnson syndrome/toxic epidermal necrolysis can shed light on novel therapeutic strategies: a comprehensive review.

Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN) are severe cutaneous adverse drug reactions (SCARs) characterized by widespread epithelial detachment and blistering, which affects the skin and mucocutaneous membranes. To date, therapeutic interventions for SJS/TEN have focused on systematic suppression of the inflammatory response using high-dose corticosteroids or intravenous immunoglobulin G (IgG), for example. No targeted therapies for SJS/TEN currently exist. Though our understanding of the pathogenesis of SJS/TEN has advanced from both an immunological and dermatological perspective, this knowledge is yet to translate into the development of new targeted therapies. Greater mechanistic insight into SJS/TEN would potentially unlock new opportunities for identifying or repurposing targeted therapies to limit or even prevent epidermal injury and blistering.

O20 Dissecting junctional epidermolysis bullosa heterogeneity

Abstract Introduction and aims Epidermolysis bullosa (EB) is a family of rare, incurable, inherited disorders that are characterized by extreme skin and mucosal fragility. One of the most severe forms is junctional EB (JEB). JEB is caused by mutations in genes encoding the skin basement membrane proteins laminin 332 (α3, β3, γ2), type XVII collagen or integrin α6β4. Despite similarities between the disease pathology of JEB and other forms of EB there is a dramatic reduction in life expectancy for patients with JEB. Patients with JEB rarely survive beyond the first year of life. To investigate this disparity, we have optimized a previously published conditional tamoxifen induced knockout JEB mouse model (Lama3flx/flxK14CreERT). This mouse model recapitulates all key features of JEB, such as thickening of the epidermis and visible blistering owing to the loss of laminin α3 at the basement membrane. Methods Single-cell (sc)RNA-Seq analysis was conducted on pooled skin samples from a healthy (control) and JEB mouse model (n = 3 per genotype). Immunofluorescent staining was conducted to confirm scRNA-Seq analysis including markers for skin differentiation (K14, transglutaminase 1, loricrin), proliferation (Ki67), basement membrane (laminin α3, laminin β3, laminin γ2, collagen VII) and immune markers (F4/80, Ly6-G/C, CD3, CD79). Results We can see large differences in cellular populations across the two conditions. With increased T- and B-immune-cell subpopulations in the JEB model compared with the healthy control. Utilizing CellChatDB we identified disease-specific signalling pathways that identify activated T- and B-cell populations interacting with JEB macrophage populations via proinflammatory pathways. Conclusions We have utilized an in vivo mouse model to gain insight into the molecular and cellular differences in a JEB diseased phenotype. Identifying changes in cellular pathways could be used as a targeted approach to identify a beneficial therapeutic treatment for JEB.

O22 Unravelling the pathophysiology of KLICK syndrome to identify therapeutically targetable pathways

Abstract Introduction and aims Keratosis linearis with ichthyosis congenita and sclerosing keratoderma (KLICK) syndrome is an ultrarare genodermatosis associated with a homozygous pathogenic variant, c.-95delC in POMP, which encodes for proteasome maturation protein. Clinical manifestations include linear hyperkeratotic papules in the flexures, palmoplantar keratoderma and pseudoainhum. The pathophysiology of KLICK syndrome is poorly understood, resulting in nonspecific, limited treatment options. The aim of this work is to characterize KLICK syndrome at the cellular and molecular level, with the utilization of transcriptomic techniques for potential drug repurposing. Methods A lesional skin biopsy was obtained, following informed consent, from a 32-year-old female patient with KLICK syndrome. Histology and immunostaining were used to characterize the disrupted skin architecture. RNA sequencing was used to identify key dysregulated pathways and the L1000FWD reverse transcriptomics tool enabled the generation of a shortlist of potential therapeutics for KLICK syndrome. CRISPR editing techniques and induced pluripotent stem cell (iPSC) reprogramming Methods were used to generate cellular based models carrying the pathogenic POMP variant in order to support evaluation of the shortlisted therapeutic options. Results KLICK syndrome skin histology featured hyperkeratosis, hypergranulosis and delayed cellular flattening. Consistent with pathological features, pathway analysis of transcriptomic data revealed keratinocyte differentiation and epidermis development as key upregulated pathways in KLICK syndrome. Interestingly, the Panther pathway analysis tool suggested dysregulation in epidermal growth factor receptor (EGFR) signalling, and reverse transcriptomics identified erlotinib, an EGFR inhibitor, as a potential therapeutic treatment for KLICK syndrome. HaCaT keratinocytes were successfully edited using CRISPR-Cas9 to carry the KLICK patient POMP variant and a KLICK patient iPSC line was generated to support in vitro KLICK syndrome modelling. Conclusions KLICK syndrome is a hyperproliferative inflammatory skin disorder associated with a pathogenic variant in POMP. Reverse transcriptomics is a useful tool for shortlisting drugs with biologically relevant mechanisms of action with potential for treatment of rare genetic diseases such as KLICK syndrome.

Individualized strategies for depression: narrative review of clinical profiles responsive to vortioxetine.

Depression is a highly heterogeneous disorder, often resulting in suboptimal response and remission rates. This underscores the need for more nuanced clinical characterization of patients to tailor individualized treatment plans. Emerging evidence highlights the critical role of cognitive and emotional dysfunction in major depression, prompting the exploration of novel therapeutic interventions that target these specific symptom domains. Vortioxetine, a multimodal antidepressant, enhances serotonergic activity while also modulating several other neurotransmitter systems involved in depressive symptoms such as emotional blunting, anhedonia, and cognitive dysfunction. Numerous randomized, placebo-controlled trials have demonstrated vortioxetine's efficacy and safety in treating depression, particularly in specific subgroups of depressed patients, including those with cognitive deficits and comorbid anxiety symptoms or disorders. Although not randomized or placebo-controlled, studies have also shown vortioxetine's efficacy in depressed patients with emotional blunting or anhedonia. Vortioxetine's ability to effectively treat a range of depressive symptoms, including anhedonia, emotional blunting, anxiety, and cognitive dysfunction, provides an individualized treatment solution for depressed individuals suffering from these symptoms. The purpose of this paper is to identify clinical profiles of patients who may benefit from vortioxetine, with the goal of optimizing therapeutic outcomes. Vortioxetine has been shown to be effective for patients with depression and symptoms such as anhedonia, emotional blunting, anxiety, and cognitive dysfunction. Tailoring treatment plans to individual needs and personalizing treatment choices based on the specific symptoms presented by depressed patients improve treatment outcomes.

Molecular Insights of Drug Resistance in Epilepsy: Multi-omics Unveil.

Epilepsy is a devastating neurological disorder mainly associated with impaired synchronic discharge that leads to sensory, motor, and psychomotor impairments. Till now, about 30 anti-seizure medications (ASMs) have been approved for the management of epilepsy, yet one-third of individuals still have uncontrollable epilepsy and develop resistance. Drug resistance epilepsy (DRE) is defined as the condition where two ASMs fail to control the seizure in epileptic patients. The leading cause of the resistance was the extended use of ASMs. According to various studies, alterations in some genes and their expressions, along with specific metabolic impairments, are suggested to be associated with ASMs resistance and DRE pathophysiology. Several factors aid in the pathophysiology of DRE, such as alterations in protein-encoding genes such as neurotransmitter receptors, drug transporters, ion channels, and drug targets. Furthermore, the altered metabolite levels of metabolites implicated in neurotransmitter signaling, energetic pathways, oxidative stress, and neuroinflammatory signaling differentiate the epileptic patient from the DRE patient. Various DRE biomarkers can be identified using the "integrated omics approach," which includes the study of genomics, transcriptomics, and metabolomics. The current review has been compiled to understand the pathophysiological mechanisms of DRE by focusing on genomics, transcriptomics, and metabolomics. An effort has also been made to identify the therapeutic targets based on identifying significant markers by a multi-omics approach. This has the potential to develop novel therapeutic interventions in the future.

De Novo Elastin Assembly Alleviates Development of Supravalvular Aortic Stenosis.

A series of incurable cardiovascular disorders arise due to improper formation of elastin during development. Supravalvular aortic stenosis (SVAS), resulting from a haploinsufficiency of ELN, is caused by improper stress sensing by medial vascular smooth muscle cells, leading to progressive luminal occlusion and heart failure. SVAS remains incurable, as current therapies do not address the root issue of defective elastin. We use SVAS here as a model of vascular proliferative disease using both human induced pluripotent stem cell-derived vascular smooth muscle cells and developmental Eln± mouse models to establish de novo elastin assembly as a new therapeutic intervention. We demonstrate mitigation of vascular proliferative abnormalities following de novo extracellular elastin assembly through the addition of the polyphenol epigallocatechin gallate to SVAS human induced pluripotent stem cell-derived vascular smooth muscle cells and in utero to Eln± mice. We demonstrate de novo elastin deposition normalizes SVAS human induced pluripotent stem cell-derived vascular smooth muscle cell hyperproliferation and rescues hypertension and aortic mechanics in Eln± mice, providing critical preclinical findings for the future application of epigallocatechin gallate treatment in humans.

Contemporary interventions in counselling children and adolescents: A literature review

This study explores the evolving landscape of therapeutic interventions for children and adolescents in contemporary society. It examines the growing body of research on practical counselling approaches tailored to address the unique challenges young people face today. The review acknowledges the importance of considering developmental stages when selecting interventions and emphasizes the need for culturally competent practices. Mental health concerns among children and adolescents are on the rise. Factors such as social media pressures, academic stress, and family dysfunction contribute to a complex landscape of emotional and behavioural issues. Effective therapeutic interventions are crucial to supporting the mental wellbeing of this population. By examining contemporary counselling approaches, this review aims to equip mental health professionals with the knowledge and tools to provide the most effective care for their young clients. This review employs a systematic search of academic databases and peer-reviewed journals, focusing on publications within the past five years. Additionally, relevant books and chapters by leading authorities in the field will be incorporated. This review is expected to identify a range of effective contemporary interventions for counselling children and adolescents. It will highlight the importance of tailoring interventions to address specific needs, developmental stages, and cultural backgrounds. The review may also identify gaps in the current research and suggest areas for future investigation. By providing a comprehensive overview of contemporary interventions in counselling children and adolescents, this review aims to contribute to the advancement of evidence-based practices in the field. Equipping mental health professionals with effective tools will ultimately contribute to improved outcomes and enhanced wellbeing for young people facing mental health challenges.

Transcranial direct current stimulation in the treatment of alcohol, tobacco and opioid use disorder in clinical studies

Transcranial direct current stimulation (tDCS) is a promising research tool to address substance abuse, including alcohol, tobacco, opioid, and drug use disorders. The present literature review compared previous studies conducted with various current intensities, application regions, durations of stimulations, and different region targets of the brain. Studies based on the analyses conducted after tDCS administration in substance use disorder were promising for the use of tDCS as adjunctive therapy to reduce visible psychological and neurological symptoms of the addiction. Therefore, we aimed to provide an insight into the current state of research on tDCS as a therapeutic intervention in substance use disorders, identify gaps in the literature, and emphasize future investigation areas. Ultimately, the review sought to contribute to the understanding of the role of tDCS in addressing the complex challenges posed by substance use disorder, and its potential as a complementary or adjunctive treatment modality in addiction care. The present study identified that the left dlPFC and brain regions were effective targets for 1 mA and 2 mA tDCS current density in tobacco/nicotine use disorder. Also, the left dlPFC and 2 mA current density were identified as effective targets for tDCS in alcohol use disorder. Furthermore, left dlPFC and 2 mA current density were identified as effective targets for tDCS in opioid use disorder. Additionally, the right/left dlPFC, orbital frontal cortex, thalamus, and 2 mA current density were identified as effective targets for tDCS in other drug or substance use disorders. Animal studies demonstrated that tDCS was promising in reducing neuropathic pain, modulating neuropeptide Y activity, and reducing the redevelopment of ethanol consumption in animal models. However, further research is required to fully understand the optimal tDCS application parameters.

HEART FAILURE AND ITS PHENOTYPES IN THE REAL WORLD: DATA FROM THE CARDIOVASCULAR OBSERVATORY OF FRIULI–VENEZIA GIULIA

Abstract Introduction Heart Failure (HF) is a disease with high mortality and morbidity burden. Guidelines classify HF according the ejection fraction (EF) in three phenotypes. HF with preserved ejection fraction (HFpEF) is the most prevalent in the real world for increasing in life expectancy and the presence of many comorbidities. Aim Knowing and analysing the epidemiology and clinic features of HF to create the most appropriate diagnostic–therapeutic pathways. The Cardiovascular Observatory of Friuli–Venezia Giulia (CVOFVG), which allows this analysis, is a powerful tool for clinical governance. Results In Friuli Venezia Giulia, at 1st January 2023, 21.212 alive patients had a HF diagnosis (1.78% prevalence). Considering the different phenotypes, in the real world, from 2018 to 2023, the most prevalent patients had EF > 40% (in 2022: HFpEF 55%, HFmrEF 9% and HFimpEF 20%). In our registry patients with HFpEF are older (mean age 80±10 yrs vs 79±11 yrs in HFmrEF vs 76±11 yrs in HFrEF), mostly women (52% vs 38% in HFmrEF, vs 27% in HFrEF), with many cardiovascular (CV) diseases, such as atrial fibrillation (57% vs 53% in HFmrEF vs 49% in HFrEF) and non CV as obesity and chronic obstructive pulmonary disease. As expected, ischemic heart disease is prevalent in HFrEF in 59% vs 38% in HFpEF. In the CVOFVG, in 2022 the hospitalization rate was 1260/100000 inhabitants (262 for HF hospitalization, 188 for CV causes, 808 for non CV disease). HFrEF has a higher incidence of HF hospitalization (54.3 on 166.2/100.000 totals). Furthermore, HFpEF because of its high prevalence, has a higher rate of CV and HF hospitalization than HFrEF (77.8 vs 54.3/100.000 inhabitants for year, 929 hospitalization vs 646). The mortality in one year is higher in HFrEF and HFmrEF vs HFpEF and above all HFimpEF. Drug prescription was different from clinical trial: in 2022, 72.2% of HFrEF patients received renin angiotensin system inhibitors (RASi), 89.2% beta blockers (BB), 62.6% mineralocorticoid receptor antagonist (MRA) and 44% SGLT2–inibitors (SGLT2–I). Same prescription for RASi, BB, MRA was in HFimpEF and HFmrEF, less for HFpEF. Conclusions The patients’ complexity, their increasing number, the new drugs for HFpEF lead to create multidisciplinary pathways, above all for HFpEF patients who are frequently visited by General Physicians, Internal Physicians and Geriatrics, to select the most complex case where a therapeutic intervention can be efficacy.

Power dynamics in couple relationships: A review and applications for systemic family therapists.

Power dynamics, generally defined as the patterns of partners enacting or resisting influence, are inherent in all relationships. Power structures and processes play a role in people's perceptions of themselves and others, their feelings and emotions, and both their implicit and explicit behaviors. As such, understanding power dynamics is crucial for fully conceptualizing and intervening within relationships. Although power was not always given high priority in many of the early systemic family therapy models, that has changed over the years, with scholars working to address how power is manifested in relationships, how power imbalances affect relationships, and how power can be addressed more explicitly in treatment. Nonetheless, there is much additional work needed to ensure that systemic therapists have an appropriate depth of understanding regarding power dynamics to fully recognize their manifestations in relationships and then intervene appropriately. To help in these efforts, this paper aims to synthesize relational power research into a more complete description of what power is and how it is enacted in couple relationships. To do this, we introduce relevant perspectives of power not fully integrated with family therapy theories. Overall, we provide a brief history of power-oriented research in the fields of family therapy, outline couple research regarding the sources of power that can inform therapeutic case conceptualizations and interventions, describe how power is addressed in specific couple therapy models, and highlight some important clinical applications that can help systemic therapists more fully address power.

THE IMPACT OF NURSE–LED EDUCATION, ON QUALITY OF LIFE IN PATIENTS WITH HEART FAILURE DURING CARDIAC REHABILITATION: A REVIEW OF THE LITERATURE

Abstract Introduction The complex mosaic of cardiac rehabilitation (CR) in heart failure (HF) is composed of several "pieces": adherence to the rehabilitation program (Mazza & Paneroni, 2019), therapeutic education, improvement of perceived QoL and self–care (Deek et al., 2017). To improve the patient‘s QoL, however, it is imperative to position them at the focal point of the care process. Several studies, have documented the favorable impact of CR on QoL in HF patients (Conway et al., 2015). Although variations in therapeutic and rehabilitative treatment plans vary according to each stage of the disease, adopting lifestyle changes is a recommended strategy regardless of the stage of HF from which the person is affected (ACC/AHA 2005; Yancy et al., 2005). However, despite the evidence–based benefits of post–discharge interventions combined with patient education in HF patients, the specific impact and benefits of nurse–led patient education alone, independent of the broader health care system, remain still unclear. Objective This review aims to investigate the influence of the nurse‘s educational role on the quality of life of patients with HF in the setting of cardiac rehabilitation. Materials and Methods A comprehensive literature review was conducted using databases such as PubMed, Scopus, CIHNAL Complete and EMBASE. The review followed the PICO methodology, with the formulation of a research question. MeSH terms and keywords were combined using Boolean operators and inclusion criteria. The primary outcome of interest was improved QoL. Relevant articles were subjected to preliminary analysis on the Rayyan platform. They were first evaluated by title and abstract, then selected for full–text reading, and finally subjected to "quality appraisal". Results, Discussion and Conclusions Individuals with HF experience a decline in QoL and are at increased risk of poor prognosis, characterized by exacerbation episodes leading to frequent re–hospitalizations and increased mortality rates (Heo et al., 2009; Rechenberg et al., 2020; Freedland et al., 2021). Therefore, improving QoL in patients with HF is a priority from both care and organizational perspectives (Ades et al., 2013; Darkins et al., 2015). Notably, a tailored CR educational program helps to improve self–care management and, consequently, improve QoL. That highlights the importance of nurse–led CR for HF patients in improving health behaviors and adherence to therapeutic practices.

Analysis of tumor microenvironment composition in vestibular schwannomas: insights into NF2-associated and sporadic variations and their clinical correlations

ObjectiveVestibular schwannomas (VS), benign tumors stemming from the eighth cranial nerve’s Schwann cells, are associated with Merlin gene mutations, inflammation, and the tumor microenvironment (TME), influencing tumor initiation, maintenance, and potential neural dysfunction. Understanding TME composition holds promise for systemic therapeutic interventions, particularly for NF2-related schwannomatosis.MethodologyA retrospective analysis of paraffin-embedded tissue from 40 patients (2013-2020), evenly divided by neurofibromatosis type 2 status, with further stratification based on magnetic resonance imaging (MRI) progression and hearing function. Immunohistochemistry assessed TME components, including T-cell markers (CD4, CD8, CD25), NK cells (CD7), and macrophages (CD14, CD68, CD163, CCR2). Fiji software facilitated image analysis.ResultsT-cell markers (CD4, CD8, CD7) exhibited low expression in VS, with no significant NF2-associated vs. sporadic distinctions. Macrophage-related markers (CD14, CD68, CD163, CCR2) showed significantly higher expression (CD14: p = 0.0187, CD68: p < 0.0001, CD163: p = 0.0006, CCR2: p < 0.0001). CCR2 and CD163 significantly differed between NF2-associated and sporadic VS. iNOS, an M1-macrophage marker, was downregulated. CD25, a regulatory T-cell marker, correlated significantly with tumor growth dynamics (p = 0.016).DiscussionImmune cells, notably monocytes and macrophages, crucially contribute to VS pathogenesis in both NF2-associated and sporadic cases. Significant differences in CCR2 and CD163 expression suggest distinct immune responses. Regulatory T-cells may serve as growth dynamic markers. These findings highlight immune cells as potential biomarkers and therapeutic targets for managing VS.

Rapid and Reliable Quantification of Urinary Malondialdehyde by HILIC-MS/MS: A Derivatization-free Breakthrough Approach

BackgroundThe development of fast analytical methods is crucial for the research, discovery, and confirmation of crucial biomarkers. Furthermore, the implementation of fast analytical strategies contributes to efficient and time-effective procedures. In this sense, analysis of malondialdehyde (MDA) has become an important tool for understanding the role of oxidative stress in various diseases and for evaluating the efficacy of therapeutic interventions. ResultsA rapid and robust liquid chromatography tandem mass spectrometry method (HPLC-MS/MS) has been developed to determine endogenous amounts of malondialdehyde (MDA) in human urine without any associated derivatization reaction. MDA was separated in 4 minutes through a Urea-HILIC column and was analyzed using a triple quadrupole mass spectrometer in negative electrospray ionization mode. With a 50-fold dilution as the only sample pretreatment after alkaline hydrolysis, no matrix effect was present, which allowed for a fast and simple quantification by means of an external standard calibration with a limit of detection of 0.20 ng mL-1. The whole methodology was validated by analyzing unspiked and spiked urine samples from ten healthy individuals and comparing with the results obtained by the standard addition method. MDA was detected in all cases, with natural concentrations varying from 0.11±0.03 to 0.31±0.03 mg g-1 creatinine. Accuracies were found to be satisfactory, ranging from 95% to 101%. The proposed method also exhibited good repeatability and reproducibility (RSD<15%) for four quality control levels. SignificanceThe main significance of this method is the avoidance of a derivatization reaction for the determination of urinary MDA, this constituting a step forward when compared with previous literature. This breakthrough not only streamlines time analysis to less than five minutes per sample but also results in a more robust procedure. Consequently, the method here developed could be applied to subsequent future research involving the determination of MDA as a lipid peroxidation biomarker, where simple, rapid, and reliable methods could represent a significant improvement.

NEMO/NF-κB signaling functions as a double-edged sword in PanIN formation versus progression to pancreatic cancer

BackgroundPancreatic ductal adenocarcinoma (PDAC) is marked by a dismal survival rate, lacking effective therapeutics due to its aggressive growth, late-stage diagnosis, and chemotherapy resistance. Despite debates on NF-κB targeting for PDAC treatment, no successful approach has emerged.MethodsTo elucidate the role of NF-κB, we ablated NF-κB essential modulator (NEMO), critical for conventional NF-κB signaling, in the pancreata of mice that develop precancerous lesions (KC mouse model). Secretagogue-induced pancreatitis by cerulein injections was utilized to promote inflammation and accelerate PDAC development.ResultsNEMO deletion reduced fibrosis and inflammation in young KC mice, resulting in fewer pancreatic intraepithelial neoplasias (PanINs) at later stages. Paradoxically, however, NEMO deletion accelerated the progression of these fewer PanINs to PDAC and reduced median lifespan. Further, analysis of tissue microarrays from human PDAC sections highlighted the correlation between reduced NEMO expression in neoplastic cells and poorer prognosis, supporting our observation in mice. Mechanistically, NEMO deletion impeded oncogene-induced senescence (OIS), which is normally active in low-grade PanINs. This blockage resulted in fewer senescence-associated secretory phenotype (SASP) factors, reducing inflammation. However, blocked OIS fostered replication stress and DNA damage accumulation which accelerated PanIN progression to PDAC. Finally, treatment with the DNA damage-inducing reagent etoposide resulted in elevated cell death in NEMO-ablated PDAC cells compared to their NEMO-competent counterparts, indicative of a synthetic lethality paradigm.ConclusionsNEMO exhibited both oncogenic and tumor-suppressive properties during PDAC development. Caution is suggested in therapeutic interventions targeting NF-κB, which may be detrimental during PanIN progression but beneficial post-PDAC development.Graphical

Can Stem Cell Therapy be an Effective Therapeutic Option for Complex and Neurological Conditions?

This article explores the feasibility and potential benefits of stem cell therapy as a promising therapeutic intervention for individuals facing severe behavioral and cognitive impairments, targeting underlying neurological mechanisms to offer potential symptom improvement and function restoration, thereby enhancing the quality of life for patients and potentially alleviating stress and burnout among caregivers. The aims of the study include investigating the efficacy of stem cell therapy in addressing these impairments and evaluating its potential to alleviate caregiver burden. Through a comprehensive review of existing literature, clinical trials, and studies, this research synthesizes the mechanisms of stem cell therapy, potential applications, clinical trial outcomes, caregiver experiences, and economic implications. The methods employed involve analyzing diverse data sources to provide a holistic understanding of the possibilities and challenges associated with stem cell therapy. Key outcomes of the study include highlighting promising results from clinical trials targeting conditions such as autism spectrum disorder, Parkinson's disease, and Crohn's disease, which underscore the transformative potential of stem cell therapy in patient care. Additionally, economic implications suggest potential reductions in healthcare costs and alleviation of financial strain on individuals, families, and healthcare providers. Stem cell therapy represents a paradigm shift in healthcare, promising to redefine treatment approaches and alleviate broader societal burdens associated with complex neurological conditions. The relevance of the study lies in emphasizing the need for continued research, ethical considerations, and multidisciplinary collaboration to ensure the safe and effective integration of stem cell therapy into clinical practice, ultimately offering new hope and improved care for patients, caregivers, and healthcare systems worldwide.

ACUTE MYOCARDIAL INFARCTION WITH “ST” SEGMENT ELEVATION IN SOUTHERN MARCHE: INTRA AND POST–HOSPITAL PROCESS INDICATORS IN THE APPLICATION OF THE UNIFIED PDTA “RETE STEMI” AST OF ASCOLI PICENO AND FERMO. LONGITUDINAL OBSERVATIONAL CLINICAL STUDY

Abstract Background SCAs (Acute Coronary Syndromes) are one of the most frequent cardiovascular diseases, among which STEMI is certainly the most relevant in terms of short and long–term prognosis. The latest ESC Guidelines about ACS–STEMI indicate some key elements in terms of diagnosis and treatment, from first contact to cardiological rehabilitation. Objective Detect the number of SCA–STEMIs managed in the AST of Ascoli Piceno, along with intra and post–hospital process indicators provided in the PDTA of the “RETE STEMI” issued by the Marche Region specifically in AST of Ascoli Piceno. MATERIALS &amp; METHODS An observational, longitudinal, monocentric, not–for–profit clinical study without diagnostic–therapeutic interventions was designed. The data collection started on 01/11/2022 and ended on 30/06/2023, at “Madonna del Soccorso” Hospital of San Benedetto del Tronto and “Mazzoni” Hospital of Ascoli Piceno in Cardiological Intensive Care Units (UTIC). Useful data for this study were acquired every month, consulting the folders of each patient diagnosed with “SCA–STEMI”, and then, reported in an Excel sheet, they were processed with different mathematical and statistical functions, including frequency, percentage, arithmetic average and standard deviation, and later used to design tables and graphs. All the data have been aggregated in full compliance with current legislation on the protection of patient privacy. Results The most significant results are related to the time between the first medical contact and the’ starting of the PPCI procedure, with an average time detected of 110 minutes. Another very important result is the number of patients sent to a post–acute cardiological rehabilitation program (PCRP): from the present study it appears that only the 1% of the entire sample was sent to a PCRP. Finally, other indicators considered in relation to the PDTA appear to be in line with the current Guidelines. Conclusions The present study highlights the aspects of the STEMI PDTA of the Marche Region which could be improved, namely reducing FCM–Wire crossing time and sending patients to a cardiological rehabilitation program, also by implementing the figure of the nurse specialized in the critical and cardiological area.