Vol 5, No 1 (2026)

Aim. Perivascular adipose tissue (PVAT) is widely recongnized as a metabolically active organ that possibly play a role in the development of atherosclerotic cardiovascular (CV) disease. Accordingly, PVAT thickness at different arterial districts could serve as a marker of atherosclerosis and CV risk. The aims of our study were 1) to evaluate the feasibility a non-invasive, ultrasound-based approach for measuring carotid PVAT thickness (cPVATt), and 2) to evaluate the association between cPVATt, carotid atherosclerosis burden, and CV risk.
Methods. We conducted an observational, cross-sectional, pilot study. Carotid PVAT was evaluated bilaterally by ultrasonography using a 10-MHz multifrequency linear probe positioned at the base of the neck in contact with clavicle, perpendicular to the skin and in a transverse orientation. The mean distance between the common carotid adventitia and the sternocleidomastoid muscle anteriorly and the longus colli muscle posteriorly, measured on both the right and the left sides, was used as a cumulative measure of cPVATt.
Results. A total of 465 patients were included in the study. The median value of cPVATt was 0.68 (0.58-0.85) cm. Significant direct correlations emerged between cPVATt and body mass index (r=0.170, p<0.001), waist circumference (r=0.224, p<0.001), neck circumference (r=0.269, p<0.001), uric acid (r=0.106, p=0.031), triglycerides (r=0.095, p=0.048), and hs-CRP (r=0.202, p=0.019). A significant inverse correlation was observed between cPVATt and HDL cholesterol (r=-0.152, p=0.001). No correlation was observed between cPVATt and any measure of carotid atherosclerotic burden. There was a significant increase in cPVATt across CV risk categories (p for trend=0.043).
Conclusions. The present study preliminarly demonstrates the feasibility of an ultrasound approach for assessing cPVAT, the reliability of cPVATt as a measure of adiposity and its potential value as a marker of increased CV risk. However, it does not show any significant correlation between cPVATt and carotid atherosclerotic burden.

Aim Heart failure (HF) is a clinical syndrome involving structural and/or functional cardiac abnormalities, classified as reduced (HFrEF) or preserved (HFpEF) based on the ejection fraction percentage of the left ventricle. As extracellular vesicles (EVs) reflect onset and severity of cardiac diseases, they attract interest as potential liquid biopsies. Aim of the present project was to characterize EVs in HF patients, investigating their potential as biomarkers and tools to discriminate between HFrEF and HFpEF clinical phenotypes.
Methods The study included 39 HF patients (13 HFpEF and 26 HFrEF) and 28 volunteers (CTR). EVs were isolated from plasma by size-exclusion chromatography and ultracentrifugation, then characterized using nanoparticles tracking analysis, transmission electron microscopy (TEM), Western blot (WB) and flow cytometry (FACS). Functional assays using patient-derived EVs were performed on cellular models of monocyte (THP-1) and cardiomyocyte (H9C2).
Results Diagnosis of HF relied on echocardiographic (e.g. E/e' ratio) and biochemical parameters (e.g. NT-proBNP). Isolation of EV was confirmed by FACS and WB analyses (e.g. the presence of CD63, CD9, CD81, Alix and β1 integrin), while integrity by TEM. EV size was increased in HF (nm: 202 vs 181). Among different subpopulations of EVs, those from monocytes (CD14+), macrophages (CD206+), neutrophils (CD66b+), endothelial cells (CD202b+), activated endothelial cells (CD62E+), cardiomyocytes (CD172a+), platelets (CD41a+), were significantly reduced in HF. Conversely, EVs released by T helper lymphocytes (CD4+) were significantly increased in HF patients when compared to controls. Treatment of THP-1 and H9C2 cells with EVs derived from HF patients led to an increased expression of proinflammatory cytokines (i.e. IL-1, IL-1, IL-6), when compared to cells treated with EVs isolated from CTR subjects. This change was mostly driven by EVs derived from HFpEF patients.
Conclusions EVs derived from HF patients exhibit a distinct profile that reflects the hemodynamic characteristics of the condition and possess proinflammatory properties.

Background and objectives: People living with HIV (PLWH) face an increased cardiovascular (CV) risk due to the interaction of traditional risk factors, chronic inflammation and cumulative antiretroviral therapy (ART) adverse metabolic effects. However, standard risk models often underestimate this burden, limiting effective prevention. This study evaluates LDL cholesterol target achivement in PLWH, based on European Society of Cardiology guidelines, and explores its association with clinical, immunological, and therapeutic HIV-related variables.
Methods: A retrospective analysis was conducted on 246 HIV-positive patients, aged ≥40, on ART at Niguarda Hospital. Clinical, laboratory, and therapeutic data were extracted from the hospital’s electronic registries, while ten-year CV risk was assessed using SCORE2 from which each patients LDL cholesterol target was defined.
Results: Only 27.2% of the analyzed cohort achieved the recommended LDL cholesterol targets; a significantly higher prevalence of uncontrolled profiles was observed among patients belonging to the “high” or “very high” SCORE2 risk categories (29.3 and 14.6% of the population, respectively). 35.4% of the patients take statins, 12.2% ezetimibe while only the l’11.4% take their association. Univariate analysis showed that lower value of total cholesterol (r=-0.490, p<0.0001), triglycerides (r=-0.188, p=0.003), systolic blood pressure (r=-0.190, p=0.003), and SCORE2 risk class (r=0.270, p<0.0001) were significantly associated with an increased likelihood of achieving the LDL cholesterol target, whereas no significant relation was found with HIV-specific variables.
Conclusions: LDL cholesterol target achievement in PLWH remains suboptimal. A refined predictive model integrating HIV-specific variables, could be useful to enhance individualized risk stratification and to optimize therapeutic strategies tailored to PLWH.

Aim: Chronic inflammation underlies numerous diseases, including atherosclerosis. The identification of anti-inflammatory agents, particularly from dietary sources, is of great interest for the development of functional products targeting early stages of chronic inflammatory conditions. This study investigated the in vitro anti-inflammatory activity of chiral phenyl-γ-valerolactones, the main colonic metabolites of flavan-3-ols.
Methods: Human dermal fibroblasts were treated with 10 phenyl-γ-valerolactones (1 μM) for 48 hours. Compounds included pure enantiomers and methylated or sulfated derivatives, tested at concentrations representative of plasma levels following dietary intake. During the first 24 hours, cells were treated under basal conditions; during the second 24 hours, treatments were repeated in the presence or absence of lipopolysaccharide (LPS, 1 μg/mL). Cytotoxicity was assessed by MTT and lactate dehydrogenase assays (LDH). Anti-inflammatory activity was evaluated by measuring IL-6 and IL-8 secretion using ELISA, with data normalized to protein content (bicinchoninic acid assay). To investigate the underlying mechanism of action, NF-κB activation was assessed by western blot analysis of p65 expression, normalized to β-actin. A 24-hour pharmacokinetic study was conducted to evaluate compound biotransformation and to characterize metabolic products, monitoring 31 phenyl-γ-valerolactones.
Results: None of the tested compounds induced cytotoxicity. (4R)-5-(4ʹ-hydroxyphenyl)-γ-valerolactone (R-CC01) reduced IL-6 and IL-8 secretion by 76% (p<0.001) and 70% (p<0.01), respectively, while its enantiomer (S-CC01) inhibited IL-6 by 89% (p<0.001) and IL-8 by 86% (p<0.01). (4R)-5-(3ʹ,4ʹ-dihydroxyphenyl)-γ-valerolactone (R-CC02) reduced both IL-6 and IL-8 by 83% (p<0.001). (4S)- and (4R)-5-(3ʹ-hydroxy-4ʹ-methoxyphenyl)-γ-valerolactones (CC03) reduced IL-6 by 90% and 78% (p<0.001), and IL-8 by 87% and 71% (p<0.01), respectively. Western blot analysis showed reduced NF-κB activation, with p65 levels decreased by 37% (R-CC01, p<0.05), 61% (R-CC02, p<0.01), and 73% (R-CC03, p<0.001). The analysis of cellular metabolism revealed that within 24 hours R-CC01 remained unmodified, whereas R-CC02 and R-CC03 formed sulfate metabolites in a time-dependent manner.
Conclusions: Phenyl-γ-valerolactones significantly reduced pro-inflammatory cytokine secretion in LPS-stimulated human fibroblasts, partly through NF-κB inhibition. These findings support their potential role in dietary or nutraceutical strategies targeting chronic inflammation.

Aim. The epidemiological burden MASLD have been slowly increasing in recent years. Starting from a background of metabolic dysfunction, we evaluated the associations between adipo-neuroinflammation markers (LCN2), indicators of cognitive impairment (MMSE score), surrogate cardiovascular risk indicators (RHI, IMT and MMEE), and MASLD.
Methods. In this cross-sectional study, we enrolled a group of 40 patients with a recent diagnosis of MASLD and a control group of 40 patients with no history of liver disease.
Results. Compared with the controls, patients with MASLD had higher serum levels of LCN2, lower RHI and MMEE values, and lower MMSE scores; univariate analysis also revealed that the differences between the groups in terms of heart rate, body weight, body mass index, body surface area, glycated haemoglobin, and echocardiographic variables (interventricular septal thickness, LVPWT, EF, LAVI, and E/A ratio) were statistically significant. Multinomial regression revealed that the presence of MASLD was significantly positively associated with LVPWT and LCN2, and significantly negatively associated with the RHI. With regards to assessments of cognitive impairment, the presence of MASLD was significantly negatively associated with the MMSE score. We also performed ROC curve analysis to explore the ability of RHI to predict MASLD; the results yielded an AUC of 0.826 (95% CI: 0.72–0.90; p<0.0005) at an optimal cut-off value of 1.87 (sensitivity=72.5%, specificity=90%), suggesting that the RHI can serve as a marker of endothelial dysfunction and thus as an indirect indicator of cardiovascular risk in patients with MASLD.
Conclusions. Patients with MASLD have greater cognitive impairment than controls; they also have higher serum levels of LCN-2 and greater endothelial dysfunction. These results imply that subjects with MASLD have a worse cardiovascular risk profile in addition to more pronounced cognitive impairment than controls do, thus suggesting that liver plays a greater role than simply serving as the metabolic centre.

Excess circulating fatty acids contributes to the association between metabolic disorders and hormonal alterations. Indeed, obesity and diabetes mellitus are frequently associated with functional hypogonadism characterised by reduced testosterone levels. Although a high-fat-diet is known to negatively affect testicular function, its specific impact on steroidogenesis remains unclear. Therefore, the aim of this study has been to investigate the effects of different FFAs, with or without human chorionic gonadotropin (hCG) stimulation, on steroidogenesis and apoptosis in a murine Leydig cell line (mLTC1).
mLTC1 cells were exposed with increasing concentrations of palmitate (PA) or oleate (OA), in the presence or absence of 0.2 IU/mL hCG. The expression of the Steroidogenic Acute Regulatory (STAR) protein, a key marker of steroidogenesis, was evaluated by immunoblotting. Testosterone secretion was measured by ELISA, while apoptosis was evaluated by cleaved caspase-3 (C3C) protein expression using immunoblotting.
Exposure to PA significantly reduced hCG-induced STAR protein expression in a dose-dependent manner after 6h and 15h, respectively, compared with PA-free conditions. Similarly, exposure to OA reduced STAR protein expression at all concentrations tested (0.4-1 mM). Prolonged treatment with PA (96 hours) further compromised steroidogenic capacity, leading to a reduction in testosterone production. Furthermore, PA significantly increased C3C levels. Notably, hCG stimulation during the last 6h of incubation significantly reduced PA-induced C3C levels at all concentrations.
In conclusion, both PA and OA impair hCG-induced steroidogenesis, while PA additionally promotes Leydig cell apoptosis. These findings suggest a direct detrimental role of excess FFAs on testicular function in obesity and indicate a potential protective role of hCG in preserving Leydig cell viability under conditions of metabolic stress.

Cystic fibrosis is a genetic and multisystemic disease associated with a very poor quality of life and life expectancy. The introduction of the combined treatment with elexacaftor/tezacaftor/ivacaftor (ETI, Kaftrio) has extended the life expectancy of adults with cystic fibrosis (awCF), which could have an impact on the prevalence of certain chronic diseases, such as cardiovascular (CV) diseases. In this regard, in a recent work, it was observed that treatment with Kaftrio has a heterogeneous effect on CV risk factors, ameliorating some of them, such as chronic inflammation, and worsening others. Among the relevant factors determining CV risk, the capacity of HDL to promote cholesterol efflux (HDL-CEC), one of the main functions of this class of lipoproteins, has proven to be a better predictor than plasma HDL concentrations. In addition, the serum capacity to load macrophages with cholesterol (cholesterol loading capacity, CLC) represents an index of serum lipoproteins' pro-atherogenic potential.
This work aimed to evaluate the effect of six months of therapy with Kaftrio on serum lipoprotein functions, namely HDL-CEC and serum CLC, in 16 awCF, and in 8 sex and age-matched healthy controls. HDL-CEC through the main pathways was evaluated with a radioisotopic cell-based assay in specific cell models, and serum CLC was assessed fluorimetrically in human monocyte-derived macrophages THP-1.
Concerning plasma lipid profile, after treatment with Kaftrio, total cholesterol, LDL-C, and HDL-C significantly increased as compared to baseline (p<0.001, p<0.001, and p=0.023, respectively), reaching values comparable to those of healthy controls. Regarding HDL function, HDL-CEC mediated by the transporter ABCA1 was significantly lower in awCF at baseline compared to healthy controls (-14%, p=0.0142). Treatment with Kaftrio increased ABCA1 HDL-CEC compared to awCF at baseline (+7%, p=0.02499). Similarly, ABCG1 HDL-CEC was significantly lower in awCF at baseline compared to healthy controls (-22%, p=0.0218) while significantly higher after treatment compared to not treated awCF (+18%, p=0.0255). In both cases, Kaftrio restored HDL-CEC levels to values comparable to those of healthy controls.
No significant differences were found for serum CLC in awCF compared to healthy controls. Kaftrio did not have any significant impact on this parameter, despite the increased LDL-C levels observed after treatment.
In conclusion, Kaftrio treatment had a positive impact on the functional lipid profile as it increased ABCA1 and ABCG1 HDL-CEC without negatively affecting serum CLC. All these effects may contribute to reducing the CV risk in awCF.

Background: Metabolic dysfunction–associated steatotic liver disease (MASLD) encompasses a spectrum ranging from simple steatosis to metabolic dysfunction–associated steatohepatitis (MASH) and cirrhosis. Lipoprotein(a) [Lp(a)] is a well-established atherogenic risk factor, but its relationship with MASLD and its histological severity has been poorly investigated. The aim of this study was to describe plasma Lp(a) levels across different histological stages and features of MASLD.
Methods: Eighty-eight patients enrolled in the PLINIO study (ClinicalTrials.gov Identifier: NCT04036357) who underwent liver biopsy were included. Plasma Lp(a) levels were measured using a specific ELISA kit. Patients were stratified according to histological diagnosis into simple steatosis (MASLD-SS), MASH, and cirrhosis. In a subgroup of 29 patients, immunohistochemical analysis was performed to assess hepatic Lp(a) expression on liver biopsy samples.
Results: Plasma Lp(a) levels increased progressively with histological severity (MASLD-SS: 22.1 [20.2–26.6] mg/dL; MASH: 28.7 [24.2–32.8] mg/dL; cirrhosis: 31.1 [29.4–33.1] mg/dL; p=0.001). Plasma Lp(a) correlated positively with fibrosis stage (rS=0.401, p<0.001), inflammation grade (rS=0.214, p=0.045), hepatocellular ballooning (rS=0.383, p<0.001), and NAFLD Activity Score (rS=0.410, p<0.001). In multivariable regression analysis including demographic and clinical variables, plasma Lp(a) was independently associated with LDL cholesterol (β=0.003, p=0.012) after adjustment for age, sex, ALT, and diabetes. At immunohistochemical analysis, no overall correlation was found between hepatic and plasma Lp(a) or histological features. However, after excluding cirrhotic patients, hepatic and plasma Lp(a) levels correlated significantly (rS=0.436, p=0.033). Among patients with MASH, hepatic Lp(a) expression was higher in those with more severe ballooning (p=0.043).
Conclusions: In this histologically characterized MASLD cohort, Lp(a) levels increased with disease severity and correlated with fibrosis and activity parameters. These findings suggest a potential role for Lp(a) in MASLD progression and warrant further studies to clarify its contribution to liver disease pathogenesis and cardiovascular risk in this population.

Background: Enhanced platelet activation contributes to the increased cardiovascular risk of elderly patients with atrial fibrillation. The biological mechanisms underlying this phenomenon are not fully clarified. This study investigated whether age-related increases in serum thromboxane B₂ (TxB₂) are associated with platelet cyclooxygenase-1 (Cox-1) upregulation in older individuals.
Methods: Serum levels of Cox-1 and TxB₂ were assessed in patients with atrial fibrillation enrolled between 2022 and 2023. A subset of participants underwent in vitro analyses to evaluate the inhibitory effect of aspirin on platelet TxB₂ generation and to quantify platelet Cox-1 expression across different age groups (<65 vs. ≥65 years). The relationship between Cox-1 expression and aspirin-mediated inhibition of TxB₂ was also explored. Associations were analyzed using Spearman correlation, and mediation analysis was performed to assess indirect effects.
Results: The study included 134 patients. Age showed a positive correlation with both Cox-1 expression (R = 0.42, p <0.01) and TxB₂ levels (R = 0.44, p <0.01). Additionally, Cox-1 expression was positively associated with TxB₂ concentrations (R = 0.50, p <0.01). Mediation analysis demonstrated that Cox-1 partially mediated the relationship between age and TxB₂ levels (β = 5.23, 95% CI: 2.33–8.63). In vitro experiments revealed a reduced sensitivity to aspirin in older patients, as reflected by higher IC₅₀ values for inhibition of platelet TxB₂ production (96.78 μM in ≥65 years vs. 48.92 μM in <65 years). This reduced response was accompanied by increased platelet Cox-1 expression in the elderly group. Moreover, higher Cox-1 levels were inversely correlated with aspirin-induced inhibition of platelet TxB₂ (R = −0.64, p <0.01).
Conclusions: Advancing age is associated with increased thromboxane production, driven in part by platelet Cox-1 upregulation. This alteration is accompanied by a reduced capacity of aspirin to suppress Cox-1 activity, potentially contributing to the heightened thrombotic risk observed in elderly patients.

Aim: Elevated lipoprotein(a) [Lp(a)] levels have been strongly related to cardiovascular (CV) risk. However, its association with Hypertension Mediated Organ Damage (HMOD) and CV events in the primary prevention setting remains unclear. The aim of our study is to evaluate in these patients the correlation between Lp(a) levels and: (i) heart, vessels and kidney HMOD and; (ii) CV events and all-cause mortality in a primary prevention setting.
Methods: 747 low CV risk subjects were recruited between 2009 and 2014. HMOD was assessed through Pulse Wave Velocity (PWV), carotid Intima-Media Thickness (IMT), presence of carotid plaques, Left Ventricular Hypertrophy (LVH), Ejection Fraction (EF) and glomerular filtration rate (GFR). All-cause mortality and CV events up to 2021 were retrieved by electronic health records, for a median follow-up time of 10 years (I-III quartiles 9.6-11.1).
Results: Mean age was 50.8 ± 13.0 years and 63.5% of the subjects were men. The prevalence of hypertension was 37.9%, dyslipidemia 67.2%, smoking 17.8%, and diabetes mellitus 8.7%. Median Lp(a) value was 17 mg/dL (5.9–56.0), and 26.5% of patients had values above 50 mg/dL. Regarding HMOD, 10.3% subjects had arterial stiffness, 7.2% increased IMT, 19.8% carotid plaques while only 0.7% had LVH. No significant correlation was found between Lp(a) levels and indices of subclinical HMOD. Furthermore, no relationship was found between CV events and all-cause mortality and Lp(a) levels.
Conclusions: In this primary prevention cohort, elevated Lp(a) levels were not associated with significant structural damage to the heart, carotid arteries, or increased aortic stiffness and were not associated with CV events and all-cause mortality.