The MMP-9 promoter genetic variant rs3918242, mRNA and protein expression in advanced carotid plaque tissue.

BACKGROUND: The MMP-9 is a known player in atherosclerosis, yet associations of the MMP-9 -1562 C/T variant (rs3918242) with various atherosclerotic phenotypes and tissue mRNA expression are still contradictory. This study aimed to investigate the MMP-9 -1562 C/T variant, its mRNA and protein expression in carotid plaque (CP) tissue, as a risk factor for CP presence and as a marker of different plaque phenotypes (hyperechoic and hypoechoic) in patients undergoing carotid endarterectomy. The MnSOD as an MMP-9 negative regulator was also studied in relation to CP phenotypes. METHODS AND RESULTS: Genotyping of 770 participants (285 controls/485 patients) was done by tetra-primer ARMS PCR. The MMP-9 mRNA expression in 88 human CP tissues was detected by TaqMan® technology. The protein levels of MMP-9 and MnSOD were assessed by Western blot analysis. The MMP-9 -1562 C/T variant was not recognized as a risk factor for plaque presence or in predisposing MMP-9 mRNA and protein levels in plaque tissue. Patients with hypoechoic plaques had significantly lower MMP-9 mRNA and protein levels than those with hyperechoic plaque (p = 0.008, p = 0.003, respectively). MnSOD protein level was significantly higher in hypoechoic plaque compared to hyperechoic (p = 0.039). MMP-9 protein expression in CP tissue was significantly affected by sex and plaque type interaction (p = 0.009). CONCLUSIONS: Considering the differences of MMP-9 mRNA and protein expression in CP tissue regarding different plaque phenotypes and the observed sex-specific effect, the role of MMP-9 in human atherosclerotic plaques should be further elucidated.

Screening Men and Women above the Age of 50 Years for Abdominal Aortic Aneurysm: A Pilot Study in an Upper Middle Income Country.

OBJECTIVE: Screening for abdominal aortic aneurysm (AAA) is recommended in high risk populations based on local conditions. Differences in lifestyle and risk factors between countries with different income status make risk stratification based on geographic location necessary. The majority of epidemiological studies on AAA have reported data from high income countries. The aim of this study was to explore the prevalence and risk factors for AAA in an upper middle income country in Eastern Europe. METHODS: A pilot screening project for AAA, supported by a mass media campaign, was conducted in 2023 in seven cities in Serbia. Ultrasound evaluation of the abdominal aorta was performed by a registered vascular surgeon on individuals who agreed to participate. Participants who attended screening completed a questionnaire on demographic and clinical information. To assess risk factors for AAA, univariable logistic regression analysis was performed to compute the odds ratio (OR) with 95% confidence interval (CI). Multivariable logistic regression was subsequently performed with adjustments for sex, age, family history of AAA, and other relevant factors. RESULTS: A total of 4 046 participants (51.2% male and 48.8% female; mean age 68.8 ± 7.6 years) responded to the campaign. An aneurysm was found in 195 (4.8%) screened individuals (8.2% of men and 1.3% of women). In males aged 50 - 64 years, the prevalence of AAA was 5.4%. Male sex, older age, family history of AAA, being a smoker or ex-smoker, being overweight, and alcohol consumption were predictors of AAA in the univariable analysis. After adjustments in the multivariable analysis, male sex (OR 8.04, 95% CI 4.87 - 13.28), older age (OR 1.04, 95% CI 1.02 - 1.07), positive family history (OR 2.47, 95% CI 1.61 - 3.78), smoker status (OR 3.10, 95% CI 2.10 - 4.59), ex-smoker status (OR 2.13, 95% CI 1.39 - 3.27), and being overweight (OR 1.85, 95% CI 1.25 - 2.74) were independent risk factors for AAA. CONCLUSION: The prevalence of AAA has not been reduced in all countries, and screening strategies might be changed based on local epidemiological data. The results of this pilot study underline the importance of exploring the prevalence of AAA in populations with a high prevalence of smoking.

TAXINOMISIS: A cloud - based platform for risk profiling and patient specific management of the carotid artery disease.

Carotid Artery Disease is a complex multi-disciplinary medical condition causing strokes and several other disfunctions to individuals. Within this work, a cloud - based platform is proposed for clinicians and medical doctors that provides a comprehensive risk assessment tool for carotid artery disease. It includes three modeling levels: baseline data-driven risk assessment, blood flow simulations and plaque progression modeling. The proposed models, which have been validated through a wide set of studies within the TAXINOMISIS project, are delivered to the end users through an easy-to-use cloud platform. The architecture and the deployment of this platform includes interfaces for handling the electronic patient record, the 3D arterial reconstruction, blood flow simulations and risk assessment reporting. TAXINOMISIS, compared with both similar software approaches and with the current clinical workflow, assists clinicians to treat patients more effectively and more accurately by providing innovative and validated tools.Clinical Relevance - Asymptomatic carotid artery disease is a prevalent condition that affects a significant portion of the population, leading to an increased risk of stroke and other cardiovascular events. Early detection and appropriate treatment of this condition can significantly reduce the risk of adverse outcomes and improve patient outcomes. The development of a software tool to assist clinicians in the assessment and management of asymptomatic patients with carotid artery disease is therefore of great clinical relevance. By providing a comprehensive and reliable assessment of the disease and its risk factors, this tool will enable clinicians to make informed decisions regarding patient management and treatment. The impact of this tool on patient outcomes and the reduction of healthcare costs will be of great importance to both patients and the healthcare system.

A Machine Learning Model for the prediction of the progression of carotid arterial stenoses.

Atherosclerotic carotid plaque development results in a steady narrowing of the artery lumen, which may eventually trigger catastrophic plaque rupture leading to thromboembolism and stroke. The primary cause of ischemic stroke in the EU is carotid artery disease, which increases the demand for tools for risk stratification and patient management in carotid artery disease. Additionally, advancements in cardiovascular modeling over the past few years have made it possible to build accurate three-dimensional models of patient-specific primary carotid arteries. Computational models then incorporate the aforementioned 3D models to estimate either the development of atherosclerotic plaque or a number of flow-related parameters that are linked to risk assessment. This work presents an attempt to provide a carotid artery stenosis prognostic model, utilizing non-imaging and imaging data, as well as simulated hemodynamic data. The overall methodology was trained and tested on a dataset of 41 cases with 23 carotid arteries with stable stenosis and 18 carotids with increasing stenosis degree. The highest accuracy of 71% was achieved using a neural network classifier. The novel aspect of our work is the definition of the problem that is solved, as well as the amount of simulated data that are used as input for the prognostic model.Clinical Relevance-A prognostic model for the prediction of the trajectory of carotid artery atherosclerosis is proposed, which can support physicians in critical treatment decisions.

Analysis of Lower Extremity Amputations from the SerbVasc Registry.

BACKGROUND: Peripheral arterial disease (PAD) and diabetes are the major causes of lower extremity amputations (LEAs) worldwide. Morbidity and mortality in patients with LEAs are high with an associated significant burden on the global health system. The aim of this article is to report the overall morbidity and mortality rates after major and minor LEAs from the Serbian Vascular Registry (SerbVasc), with an analysis of predictive factors that influenced adverse outcomes. MATERIALS AND METHODS: SerbVasc was created in 2019 as a part of the Vascunet collaboration that is aiming to include all vascular procedures from 21 hospitals in Serbia. Prevalence of diabetes among patients with LEAs, previous revascularization procedures, the degree and the type of foot infection and tissue loss, and overall morbidity and mortality rates were analyzed, with a special reference to mortality predictors. RESULTS: In the period from January 2020 to December 2022, data on 702 patients with LEAs were extracted from the SerbVasc registry, mean age of 69.06±10.63 years. Major LEAs were performed in 59%, while minor LEAs in 41% of patients. Diabetes was seen in 65.1% of the patients, with 44% of them being on insulin therapy. Before LEA, only 20.3% of patients had previous peripheral revascularization. Soft tissue infection, irreversible acute ischemia, and Fontaine III and IV grade ischemia were the most common causes of above-the-knee amputations while diabetic foot was the most common cause of transphalangeal and toe amputations. The infection rate was 3.7%, the re-amputation rate was 5.7%, and the overall mortality rate was 6.9%, with intrahospital mortality in patients with above-the-knee amputation of 11.1%. The most significant intrahospital mortality predictors were age >65 years (p<0.001), chronic kidney disease (CKD) (p<0.001), ischemic heart disease (IHD) (p=0.001), previous myocardial revascularization (p=0.017), emergency type of admission (p<0.001), not using aspirin (p=0.041), using previous anticoagulation therapy (p=0.003), and postoperative complications (p<0.001). CONCLUSIONS: The main predictors of increased mortality after LEAs from the SerbVasc registry are age >65 years, CKD, IHD, previous myocardial revascularization, emergency type of admission, not using aspirin, using previous anticoagulation therapy, and postoperative complications. Taking into account high mortality rates after LEAs and a small proportion of previous peripheral revascularization, the work should be done on early diagnosis and timely treatment of PAD hopefully leading to decreased number of LEAs and overall mortality. CLINICAL IMPACT: Mortality after lower limb amputation from the SerbVasc register is high. A small number of previously revascularized patients is of particular clinical importance, bearing in mind that the main reasons for above-the-knee amputations were irreversible ischemia, Fontaine III and Fontaine IV grade ischemia. Lack of diagnostics procedures and late recognition of patients with PAD, led to subsequent threating limb ischemia and increased amputation rates. The work should be done on early diagnosis and timely treatment of PAD in Serbia, hopefully leading to an increased number of PAD procedures, decreased number of LEAs, and lower overall mortality.

The Value of Troponin Measurement in Carotid Revascularization: A Scoping Review.

PURPOSE: Patients undergoing carotid endarterectomy (CEA) or carotid artery stenting (CAS) are at substantially increased risk of short-term and long-term cardiac complications. Still, the role of perioperative troponin in predicting cardiac events remains unclear. The objective was to systematically summarize the existing evidence on the topic and provide directions for further research. MATERIALS AND METHODS: Studies that examined perioperative troponin values and its association with myocardial injury, and/or myocardial infarction (MI), and/or major adverse cardiac events (MACE) and postoperative mortality in exclusively CEA/CAS patients, published in English until March 15, 2022, were retrieved through a systematic search of MEDLINE and Web of Science. The study selection process was independently performed by 2 authors, while the third researcher resolved disagreements. RESULTS: Four studies with 885 participants met the inclusion criteria. Age, chronic kidney disease, presentation of carotid disease, type of closure (primary closure/venous patch/Dacron/polytetrafluoroethylene patch), coronary artery disease, chronic heart failure, and the long-term use of calcium channel blockers represent risk factors for troponin elevation, which occurred in 11% to 15.3%. Myocardial infarction and MACE occurred in 23.5% to 40%, that is, 26.5% of patients with troponin elevation, respectively, during the first 30 postoperative days. Elevated postoperative troponin levels were significantly associated with adverse cardiac events during the long-term surveillance period. The rates of cardiac-related and all-cause mortality were higher in patients with postoperative troponin elevation. CONCLUSION: Troponin measurement could be helpful in the prediction of adverse cardiac events. The predictive role of preoperative troponin, the patient population in whom routine troponin sampling should be used, and a comparison of different treatment methods/anesthesia techniques in carotid patients should be further examined. CLINICAL IMPACT: The present scoping review critically appraises the extent and nature of the existing literature data on the predictive value of troponin on the occurrence of cardiac complications in patients undergoing CEA and CAS. In particular, it provides clinicians with essential insights by systematically summarizing the core evidence and identifying knowledge gaps that may direct future research. This, in turn, may significantly alter the current clinical practice and perhaps even reduce the incidence of cardiac complications in patients undergoing CEA/CAS.

The influence of gender on 30-day adverse clinical outcomes in patients undergoing carotid surgery.

BACKGROUND: We aimed to further evaluate sex differences of perioperative and 30-day complications after carotid surgery in patients with both asymptomatic and symptomatic carotid artery stenosis. METHODS: This was a single-center prospective cohort study including 2013 consecutive patients, who were treated surgically due to extracranial carotid artery stenosis and prospectively followed. Patients who underwent carotid artery stenting and who were treated conservatively were excluded. The primary endpoints for this study were hospital stroke/transitory ischemic attack (TIA) and overall survival rates. Secondary outcomes included all other hospital adverse events, 30-day stroke/TIA, and 30-day mortality rates. RESULTS: Hospital mortality was higher in female patients with symptomatic carotid stenosis (3% vs. 0.5%, P=0.018). Bleeding requiring re-intervention occurred more often in female patients with both asymptomatic (1.5% vs. 0.4%, P=0.045) and symptomatic carotid stenosis (2.4% vs. 0.2%, P=0.022). 30-day stroke/TIA and mortality rates were higher in female patients with both asymptomatic (stroke/TIA 4.4% vs. 2.5%, P=0.041; mortality 3.3% vs. 1.6%, P=0.046) and symptomatic carotid stenosis (stroke/TIA 8.3% vs. 4.2%, P=0.040; mortality 4.1% vs. 0.7%, P=0.006). After adjusting for all confounding factors, female gender remained an important predicting factor for 30-day stroke/TIA in asymptomatic (OR=1.4, 95%CI 1.0-4.7, P=0.041) and symptomatic patients (OR=1.7, 95%CI 1.1-5.3, P=0.040), as well as for 30-day all-cause mortality in patients with asymptomatic (OR=1.5, 95%CI 1.1-4.1, P=0.030) and symptomatic carotid artery disease (OR=1.2, 95%CI 1.0-5.2, P=0.048). CONCLUSIONS: Female gender is important predicting factor for stroke/TIA and all-cause mortality, both perioperative and during the first 30 days after carotid surgery.

Treatment of vascular non-iatrogenic injuries of upper and lower extremities in tertiary vascular center.

BACKGROUND: The purpose of this study was to examine demographic and baseline characteristics of patients with vascular injuries of extremities and to define the most relevant factors which influenced an early outcome, as well as limb salvage after the management of vascular trauma. METHODS: This study used the database that included 395 patients with peripheral arterial injuries, who were treated in the tertiary vascular university center in the period between 2005-2020. Exclusion criteria were isolated thoracic, abdominal and neck injuries as well as iatrogenic injuries and injuries of intravenous addicts. Univariate binary logistic regression analysis and multiple logistic regression were used to determine risk factors for lomb loss (after vascular reconstruction) or mortality. Decision to perform primary amputation (without vascular reconstruction) was based on surgeons' preference and experience. RESULTS: Out of 395 vascular injuries treated in the period 2005-2020, 210 (53.2%) presented with non-iatrogenic vascular injuries of upper and lower extremities were analyzed. According to the univariate regression analysis, hemorrhage as the main clinical manifestation on admission (P=0.035) and early reintervention (P=0.048) increased, while an early patency of repaired artery (0.010) significantly decreased a 30-day amputation rate. Multivariate logistic regression analysis of these three variables showed that only early patency of repaired artery significantly decreased the early amputation rate (P=0.009). CONCLUSIONS: Based on presented experience, the patency of vascular reconstruction plays a crucial role in limb salvage in patients with non-iatrogenic peripheral vascular injuries. All factors that might influence the patency should be in focus of improvement.

Arterial thoracic outlet syndrome: a 30-year experience in a high-volume referral center.

BACKGROUND: The purpose of this study was to assess the clinical presentation and contemporary management of arterial thoracic outlet syndrome (TOS) in high-volume referral center. METHODS: We conducted a retrospective review of a prospectively maintained database of patients with TOS of any etiology between January 1st 1990 and 2021. Supra-, or combined supra-/infraclavicular approaches have been used for decompression/vascular reconstructions. The group was divided into two equal time periods: period 1 (1990-2006, N.=27) and period 2 (2006-2021, N.=36). RESULTS: Sixty-three consecutive patients underwent surgical treatment due to arterial TOS. Period 2 had more patients who were asymptomatic (N.=16, 44.4% vs. N.=0, 0%, P<0.001) and those presenting with critical hand ischemia (N.=12, 33.3% vs. N.=0, 0%, P=0.01), while acute limb ischemia was more common in period 1 (N.=16, 59.2% vs. N.=5, 13.9%, P<0.001). SA compression without lesion was more common in period 2 (N.=16, 44.4% vs. N.=0, 0%, P<0.001), while SA intimal damage with mural thrombus formation was more common in the period 1 (N.=12, 44.4% vs. N.=1, 2.7%, P<0.001). Decompression as an isolated procedure was performed in 25.4% (N.=16) of all asymptomatic patients, while combined decompressive and vascular procedure in 71.4% (N.=45) of patients. The most common postoperative complication was pneumothorax (N.=7, 11.1%). CONCLUSIONS: The supraclavicular approach with its modifications provides adequate decompression and allows also repair or reconstruction of the SA, as well as complete additional revascularization of the upper extremity without the need for further patient repositioning. While treatment methods and early outcomes have not changed significantly over time, there has been a trend towards different clinical and SA pathomorphological presentation.

Detection of Asymptomatic Carotid Artery Stenosis through Machine Learning.

Carotid artery disease, the pathological condition of carotid arteries, is considered as the most significant cause of cerebral events and stroke. Carotid artery disease is considered as an inflammatory process, which involves the deposition and accumulation of atherosclerotic plaque inside the carotid intima, resulting in the narrowing of the arteries. Carotid artery stenosis (CAS) is either symptomatic or asymptomatic and its presence and location is determined by different imaging modalities, such as the carotid duplex ultrasound, the computed tomography angiography, the magnetic resonance angiography (MRA) and the cerebral angiography. The aim of this study is to present a machine learning model for the diagnosis and identification of individuals of asymptomatic carotid artery stenosis, using as input typical health data. More specifically, the overall model is trained with typical demographics, clinical data, risk factors and medical treatment data and is able to classify the individuals into high risk (Class 1-CAS group) and low risk (Class 0-non CAS group) individuals. In the presented study, we implemented a statistical analysis to check the data quality and the distribution into the two classes. Different feature selection techniques, in combination with classification schemes were applied for the development of our machine learning model. The overall methodology has been trained and tested using 881 cases (443 subjects in low risk class and 438 in high risk class). The highest accuracy 0.82 and an area under curve 0.9 were achieved using the relief feature selection technique and the random forest classification scheme.

Correlation Between Proteolytic Activity and Abdominal Aortic Aneurysm Wall Morphology with Intraluminal Thrombus Volume.

BACKGROUND: The aim of this study was to examine the influence of intraluminal thrombus (ILT) volume on the level of proteolytic activity and the content of abdominal aortic aneurysm (AAA) wall. METHODS: The research was designed as a cross-sectional study at the Clinic for Vascular and Endovascular Surgery, Clinical Center of Serbia in the period from April 2017 to February 2018. During this period, a total of 155 patients with asymptomatic AAA underwent open surgical treatment and 50 were included in the study based on inclusion and exclusion criteria. Before surgery, patients included in the study were examined by MRI. During the operation, samples of ILT and AAA wall were taken for biochemical analysis. RESULTS: A statistically significant correlation was found between the volume of the ILT and largest AAA diameter (ρ = 0.56; P < 0.001). The correlation of the ILT volume on the anterior wall and the concentration of MMP-9, MMP-2 and NE/ELA in the wall did not find statistical significance. Also, no statistically significant association was found between the volume of ILT and the concentration of ECM proteins (collagen type 3, elastin, proteoglycan) in the corresponding part of the wall. The association of ILT volume with MDA was also of no statistical significance. There was a positive statistical significance found in correlation of volume of ILT and catalase activity in the wall of AAA (ρ = 0.28, P = 0.049). CONCLUSIONS: The volume of ILT in the aneurysmal sac seemed not to affect the level of proteolytic activity and the content of the aneurysm wall. However, a positive correlation was found between the ILT and the catalase activity. The effect of ILT on the aneurysm wall and its role in the progression of aneurysmal disease should be examined in future studies.

Translation and validation of the Amsterdam preoperative anxiety and information scale (APAIS) in Serbia.

OBJECTIVES: Preoperative anxiety is common and might affect surgical treatment outcomes. The aim was to translate and validate the Serbian version of the Amsterdam Preoperative Anxiety and Information Scale (APAIS). METHODS: Following translation and initial evaluation, the Serbian version (S-APAIS) was administered to 385 patients. Internal consistency, construct validity, prognostic criteria validity, and concurrent validity between S-APAIS and Visual Analogue Scale for Anxiety (VAS-A) were evaluated. RESULTS: Factor analysis revealed two factors: APAIS-anesthesia (items 1, 2, 3) and APAIS-procedure (items 4, 5, 6). The whole scale, APAIS-anesthesia, and APAIS-procedure subscales showed an adequate level of internal consistency (Cronbach's αs: 0.787, 0.806, and 0.805, respectively). High concurrent validity was observed between APAIS-anesthesia and VAS-A (ρ = 0.628, p < .001). A moderate correlation was found between APAIS-procedure and VAS-A scale (ρ = 0.537, p < .001). At the cut-off point of 9, the area under the curve (AUC) of APAIS-anesthesia was 0.815 (95% CI: 0.77-0.85, p < .001). For the APAIS-procedure, AUC was 0.772 (95% CI: 0.73-0.81, p < .001) at the cut-off point of 8. CONCLUSION: The structure of S-APAIS substantially differs from the original and allows separate measurement of anesthesia- and procedure-related anxieties. S-APAIS is a comprehensive, valid, and reliable instrument for the measurement of preoperative anxiety.

Tag Variants of LGALS-3 Containing Haplotype Block in Advanced Carotid Atherosclerosis.

OBJECTIVES: Galectin-3 affects a variety of biological processes. It is encoded by LGALS-3, located in unique haplotype block in Caucasians. Most of the studies regarding the gal-3 role in atherosclerosis are focused exclusively on protein/mRNA levels. Genetic analyses of LGALS-3 are scarce. We sought to thoroughly examine the genetic background of gal-3 and to analyze tag variants that cover more than 80% variability of the LGALS-3 containing hap-block in association with carotid plaque presence (CPP). According to Tagger server, rs4040064 G/T, rs11628437 G/A and rs7159490 C/T cover 82% (r2 > 0.8) of the genetic variance of this hap-block. Our aims were to investigate possible association of rs4040064, rs11628437 and rs7159490 haplotypes with CPP in patients with advanced carotid atherosclerosis (CA) and to analyze their possible effect on LGALS-3 mRNA expression in carotid plaques. MATERIALS AND METHODS: Study group consisted of 468 patients and 296 controls. Rs4040064, rs11628437, rs7159490 and LGALS-3 mRNA expression were detected by TaqMan® technology. RESULTS: We have found that haplotype TAC was associated with the cerebrovascular insult (CVI) occurrence (OR = 1.68, 95% CI = 1.09-2.58, p = 0.02), compared to the referent haplotype. OR was adjusted for hypertension, age and BMI. TAC also showed higher, but not statistically significant, LGALS-3 expression in carotid plaques. CONCLUSIONS: Our results suggest that rs4040064, rs11628437 and rs7159490 bear no association with CPP, neither they affect LGALS-3 mRNA in carotid plaques. However, we showed a significant association of haplotype TAC with the CVI occurrence in CA patients from Serbia. Replication and validation of our results are required.