Skipskis, Vilius

Introduction: Severe manifestations of COVID-19 are frequently characterized by acute systemic inflammation, endothelial dysfunction, and a heightened predisposition toward cardiovascular and thrombotic events (1). Many patients who developed severe COVID-19 have pre-existing chronic conditions, such as cardiovascular or metabolic disorders, which are commonly associated with impaired or dysregulated immune response. SARS-CoV-2-induced hypoxia, cytokine-driven inflammation, and direct injury to epithelial and endothelial tissues can disrupt vital biological barriers, such as the intestinal epithelium and the alveolar-capillary junction, thereby facilitating the movement of microbial components into the bloodstream (1-3). The presence of bacterial components may further augment immune activation and inflammatory cascades. Emerging data suggest that bacterial DNA fragments in the bloodstream may trigger inflammatory pathways leading to vascular injury and endothelial activation (4,5). This interaction among viral infection, bacterial translocation, and impaired host immunity may underlie increased susceptibility to complications. Nevertheless, the occurrence, composition, and taxonomic diversity of circulating bacterial DNA in complicated COVID- 19 patients have not been thoroughly characterized. Aim: To use next-generation sequencing (NGS) to detect circulating bacterial DNA and characterize its taxonomic diversity and inter-individual heterogeneity in patients recovering from severe SARS-CoV-2 infection. Methods: The study was conducted at the Laboratory of Molecular Cardiology, Institute of Cardiology, Lithuanian University of Health Sciences. Venous blood samples were obtained after completion of inpatient treatment in patients with a prior episode of severe COVID-19, following acute hospitalization (n=101). Bacterial DNA was amplified by PCR targeting the 16S rRNA gene. Samples yielding detectable 16S rRNA products underwent sequencing using Oxford Nanopore Technologies (ONT, The Oxford Science Park, UK). Taxonomic classification of both 16S rRNA amplicon data and shotgun metagenomic reads was performed using the EPI2ME platform (ONT). Reads were classified at the class level. Patients from the same clinical cohort in whom bacterial DNA was not detected served as an internal comparator group. Additionally, clinical and laboratory parameters related to cardiac injury, systemic inflammation, and coagulation were assessed to provide a clinical context. Results: Circulating bacterial DNA was detected in most of the patients (76% of the total cohort, n=101). The analysis highlighted significant variation among individual microbial profiles. Taxonomic profiling demonstrated that Gammaproteobacteria and Alphaproteobacteria were the most dominant taxa, occurring in 95% of the bacterial DNA- positive samples. In comparison, Betaproteobacteria were identified in 43% of cases, while Bacilli and Actinomycetes were less frequent, appearing in 33% and 29% of the analyzed samples, respectively. Conclusions: Bacterial DNA is detectable in the blood of patients recovering from severe COVID-19 and exhibits marked interindividual diversity. The recurrent detection of Gammaproteobacteria may indicate a non-random microbial pattern; however, further studies with rigorous contamination controls are required. These results underscore the need for further research into how bacterial components might exacerbate systemic inflammation in viral infections.

Introduction Severe forms of COVID-19 are linked to significant systemic inflammation, endothelial dysfunction, and an elevated risk of thrombotic and cardiovascular complications (1). SARSCoV-2-induced hypoxia, cytokine-driven inflammation, and direct injury to epithelial and endothelial tissues can disrupt vital biological barriers facilitating the movement of microbial components into the bloodstream (1–3). Increasing evidence indicates that microbial components in the bloodstream may contribute to inflammation-related endothelial activation and vascular damage (4,5). This interaction among viral infection, bacterial translocation, and impaired host immunity may underlie increased susceptibility to complications. Nevertheless, the occurrence, composition, and taxonomic diversity of circulating bacterial DNA in complicated COVID-19 patients have not been thoroughly characterized. Aim To investigate the presence, taxonomic diversity, and variability of circulating bacterial DNA in the venous blood of patients hospitalized with severe COVID-19 infection, using nextgeneration sequencing. Methods The study was conducted at the Laboratory of Molecular Cardiology of the Institute of Cardiology at the Lithuanian University of Health Sciences. Venous blood samples were collected from 65 patients after hospitalization for severe COVID-19 infection. PCR targeting the 16S rRNA gene was employed to amplify bacterial DNA from patient samples. At the time of analysis, sequencing was completed for 21 samples, while analysis of the remaining samples is ongoing. Subsequently, microbiome profiling using Oxford Nanopore Technologies (ONT) sequencing was performed only on samples containing 16S rRNA gene sequences. Taxonomic interpretation was performed at the class level. A healthy control group was not included, as previous studies conducted in the same laboratory have not demonstrated the presence of circulating bacterial DNA in healthy individuals. Clinical and laboratory parameters related to cardiac injury, inflammation and coagulation were evaluated. Results Bacterial 16S rRNA gene sequences were detected in 20 out of 21 analyzed samples (95.2%). Gammaproteobacteria and Alphaproteobacteria were the predominant classes, detected in 95.2% of samples. Betaproteobacteria were identified in 42.9% of samples, while Actinomycetes and Bacilli were present in 28.6% and 33.3% of cases, respectively. Within one year after hospitalization, 14 of the 21 patients were diagnosed with cardiovascular conditions. Conclusions Circulating bacterial DNA is detectable in the blood of patients after hospitalization for severe COVID-19 infection, and its interpatient variability is evident. The relevance of the Gammaproteobacteria class indicates a non-random microbial DNA signal and supports further investigation into the potential role of bacterial components in systemic inflammation.

Postoperative bleeding following cardiopulmonary bypass (CPB) remains a significant challenge.Although viscoelastic testing is increasingly used, the relative contributions of fibrinogen, platelet count and clot firm-ness to blood loss remain debated. We evaluated the diagnostic accuracy of thromboelastometry (ROTEM) comparedwith platelet aggregometry (PA) and standard tests, using the Hb/kg index to quantify blood loss.

Background: Ischemic heart disease remains the leading cause of death worldwide, with coronary microvascular dysfunction (CMD) as a key complication after ST-elevation myocardial infarction (STEMI). Endothelial dysfunction contributes to CMD, impairing vascular tone and increasing inflammation. While angiotensin-converting enzyme (ACE) inhibitors and angiotensin II receptor blockers (ARBs) aid vascular health, their efficacy may improve with therapeutic ultrasound, which enhances drug delivery and endothelial response. This study explores the combined effects of ultrasound and pharmacological treatment on the ACE axis and inflammation in endothelial and renal cells. Methods: Human umbilical vein endothelial cells (HUVECs) and human renal proximal tubular epithelial cell line RPTEC/TERT1 were treated with captopril, losartan, and dexamethasone, alone or combined with low-frequency ultrasound (LFU). Cell viability and wound-healing assays assessed cellular function, while nitric oxide (NO) and reactive oxygen species (ROS) assays were used to evaluate redox signaling. Gene expression related to the ACE axis, inflammation, and vascular and renal cell function was analyzed via qPCR. Results: Captopril and losartan combined with LFU improved endothelial cell viability, wound healing, and NO production at various concentrations, whereas only losartan with LFU enhanced cell viability and wound healing in renal cells. Dexamethasone with LFU increased ROS levels and had variable effects on RPTEC/TERT1 cell survival. Gene expression analysis showed that LFU alone reduced pro-inflammatory markers VCAM-1, ICAM-1, and PTGS2 in captopril-treated HUVECs and similarly affected CYP4F2 in losartan-treated HUVECs. LFU also decreased PTGS2 expression at higher dexamethasone concentrations. In RPTEC/TERT1 cells, LFU alone did not impact SGLT2 or GGT1 expression, but captopril with LFU downregulated GGT1, and dexamethasone with LFU upregulated SGLT2 at higher concentrations. Conclusions: This study demonstrates that LFU enhances the effects of RAS inhibitors by promoting NO synthesis and reducing oxidative stress, while its combination with dexamethasone may have variable, potentially cytotoxic effects on renal cells. Gene expression patterns suggest LFU’s anti-inflammatory potential and its role in modulating drug efficacy.

The effectiveness of lipid-lowering therapies may be insufficient in high-risk cardiovascular patients and depends on the genetic variability of drug-metabolizing enzymes. Customizing statin therapy, including treatment with atorvastatin, may improve clinical outcomes. Currently, there is a lack of guidelines allowing the prediction of the therapeutic efficacy of lipid-lowering statin therapy. This study aimed to determine the effects of clinically significant gene variants of CYP2C19 on atorvastatin therapy in patients with acute coronary syndromes. In total, 92 patients with a confirmed diagnosis of ST-elevation myocardial infarction (STEMI) or non-ST-elevation myocardial infarction (NSTEMI) were sequenced for target regions within the CYP2C19 gene on the Illumina Miniseq system. The CYP2C19 poor metabolizer phenotype (carriers of CYP2C192, CYP2C194, and CYP2C19*8 alleles) was detected in 29% of patients. These patients had significantly lower responses to treatment with atorvastatin than patients with the normal metabolizer phenotype. CYP2C19-metabolizing phenotype, patient age, and smoking increased the odds of undertreatment in patients (∆LDL-C (mmol/L) < 1). These results revealed that the CYP2C19 phenotype may significantly impact atorvastatin therapy personalization in patients requiring LDL lipid-lowering therapy.

Background/objectives. To investigate the associations between ophthalmic parameters, CYP4F2 (rs2108622) and ABCA1 (rs1883025) polymorphisms and coronary artery disease, considering the accessibility, non-invasive origin of retinal examination and its possible resemblance to coronary arteries. Subjects/methods. Overall 165 participants divided into groups based on the coronary angiography results and clinical status: control group (N = 73), MI group (N = 63), 3VD (three vessel disease) (N = 24). All the participants underwent total ophthalmic examination - optical coherence tomography (OCT) and OCT angiography of the macula region were performed and evaluated. Total cholesterol, high-density lipoprotein, low-density lipoprotein and triglyceride cholesterol (Tg-C) were tested. A standard manufacturer's protocol for CYP4F2 (rs2108622) and ABCA1 (rs1883025) was used for genotyping with TaqMan probes. Results. GCL+ layer was thicker in control group vs. 3VD group (74.00; 62.67-94.67 (median; min.-max.) vs. 71.06; 51.33-78.44, p = 0.037). T allele carriers under ABCA1 rs1883025 dominant model were shown to have ticker retina and smaller foveal avascular zone in superficial capillary plexus and smaller Tg-C concentration. ABCA1 rs1883025 was associated with retinal thickness (OR = 0.575, 95% CI 0.348-0.948, p = 0.030). Univariate logistic regression showed that ABCA1 rs1883025 CT genotype is associated with decreased risk for coronary artery disease development under overdominant genetic model (OR = 0.498, 95% CI 0.254-0.976; p = 0.042) and codominant genetic model (OR = 0.468, 95% CI 0.232-0.945, p = 0.034). Conclusions. Results of this study confirmed that non-invasive methods such as OCT of eye might be used for identification of patients at risk of CAD.

The aim of this study was to determine the effect of ABCB1 genetic polymorphism and renal function on the occurrence of ticagrelor-related dyspnea.

We conducted a research study to create the groundwork for personalized solutions within a skin aging segment. This test utilizes genetic and general laboratory data to predict individual susceptibility to weak skin characteristics, leveraging the research on genetic polymorphisms related to skin functional properties. A cross-sectional study was conducted in a collaboration between the Private Clinic Medicina Practica Laboratory (Vilnius, Lithuania) and the Public Institution Lithuanian University of Health Sciences (Kaunas, Lithuania). A total of 370 participants agreed to participate in the project. The median age of the respondents was 40, with a range of 19 to 74 years. After the literature search, we selected 15 polymorphisms of the genes related to skin aging, which were subsequently categorized in terms of different skin functions: SOD2 (rs4880), GPX1 (rs1050450), NQO1 (rs1800566), CAT (rs1001179), TYR (rs1126809), SLC45A2 (rs26722), SLC45A2 (rs16891982), MMP1 (rs1799750), ELN (rs7787362), COL1A1 (rs1800012), AHR (rs2066853), IL6 (rs1800795), IL1Beta (rs1143634), TNF-α (rs1800629), and AQP3 (rs17553719). RT genotyping, blood count, and immunochemistry results were analyzed using statistical methods. The obtained results show significant associations between genotyping models and routine blood screens. These findings demonstrate the personalized medicine approach for the aging segment and further add to the growing literature. Further investigation is warranted to fully understand the complex interplay between genetic factors, environmental influences, and skin aging.

Šis išradimas yra odos polinkio į tam tikrus fenotipinius pokyčius nustatymo būdas ir atitinkamos odos priežiūros režimo parinkimas, ir priklauso matavimo ir testavimo procesų sričiai, tiksliau matavimo ir testavimo procesams, kuriuose naudojamos nukleorūgštys, sričiai. Šiame dokumente aprašomas Lietuvos populiacijai tinkamas genetinis odos būklės tyrimas ir jo vertinimo būdas, naudojant vieno nukleotido polimorfizmų analizę genuose, susijusiuose su odos antioksidacine funkcija, odos atsparumo saulės poveikiui funkcija, odos elastingumo ir jos atramine funkcija, odos drėkinimo funkcija bei odos imuninio atsako funkcija. 1. Odos polinkio į tam tikrus fenotipinius pokyčius nustatymo būdas, apimantis:- vieno nukleotido polimorfizmų nustatymą iš anksto atrinktuose vieno nukleotido polimorfizmų regionuose, kur kiekvienas šiuose regionuose esantis vieno nukleotido polimorfizmas yra genetiškai susietas su odos polinkiu į tam tikrus fenotipinius pokyčius, kur iš anksto atrinktus vieno nukleotido polimorfizmų regionus sudaro:- pirmasis iš anksto nustatytų vieno nukleotido polimorfizmų regionų pogrupis, apimantis vieno nukleotido polimorfizmų regionus, genetiškai susijusius su odos atsparumu saulės poveikiui ir esančius genuose TYR, SLC45A21 ir SLC45A22;- antrasis iš anksto nustatytų vieno nukleotido polimorfizmų regionų pogrupis, apimantis vieno nukleotido polimorfizmų regionus, genetiškai susijusius su odos elastingumu ir jos atramine funkcija ir esančius genuose MMP1 ir ELN;- trečiasis iš anksto nustatytų vieno nukleotido polimorfizmų regionų pogrupis, apimantis vieno nukleotido polimorfizmų regioną, genetiškai susijusį su odos drėkinimo funkcija ir esantį gene AQP3;- ketvirtasis iš anksto nustatytų vieno nukleotido polimorfizmų regionų pogrupis, apimantis vieno nukleotido polimorfizmų regionus genetiškai susijusius su odos antioksidacine funkcija ir esančius genuose SOD2, GPX1, CAT ir NQO1; [...].

Introduction: Persistent coronary microcirculatory dysfunction (CMD) and elevated trimethylamine N-oxide (TMAO) levels after ST-elevation myocardial infarction (STEMI) may drive negative structural and electrical cardiac remodeling, resulting in new-onset atrial fibrillation (AF) and a decrease in left ventricular ejection fraction (LVEF). Aims: TMAO and CMD are investigated as potential predictors of new-onset AF and left ventricular remodeling following STEMI. Methods: This prospective study included STEMI patients who had primary percutaneous coronary intervention (PCI) followed by staged PCI three months later. Cardiac ultrasound images were obtained at baseline and after 12 months to assess LVEF. Coronary flow reserve (CFR), and index of microvascular resistance (IMR) were assessed using the coronary pressure wire during the staged PCI. Microcirculatory dysfunction was defined as having an IMR value ≥25 U and CFR value <2.5 U. Results: A total of 200 patients were included in the study. Patients were categorized according to whether or not they had CMD. Neither group differed from the other with regards to known risk factors. Despite making up only 40.5% of the study population, females represented 67.4% of the CMD group p < 0.001. Similarly, CMD patients had a much higher prevalence of diabetes than those without CMD (45.7% vs. 18.2%; p < 0.001). At the one-year follow-up, the LVEF in the CMD group had decreased to significantly lower levels than those in the non-CMD group (40% vs. 50%; p < 0.001), whereas it had been higher in the CMD group at baseline (45% vs. 40%; p = 0.019). Similarly, during the follow-up, the CMD group had a greater incidence of AF (32.6% vs. 4.5%; p < 0.001). In the adjusted multivariable analysis, the IMR and TMAO were associated with increased odds of AF development (OR: 1.066, 95% CI: 1.018–1.117, p = 0.007), and (OR: 1.290, 95% CI: 1.002–1.660, p = 0.048), respectively. Similarly, elevated levels of IMR and TMAO were linked with decreased odds of LVEF improvement, while higher CFR values are related to a greater likelihood of LVEF improvement. Conclusions: CMD and elevated TMAO levels were highly prevalent three months after STEMI. Patients with CMD had an increased incidence of AF and a lower LVEF 12 months after STEMI.