Association between undiagnosed obstructive sleep apnea and severe course of COVID-19: a prospective observational study.

PURPOSE: Obstructive sleep apnea (OSA) is associated with many long-term health consequences. We hypothesized that previously unrecognized and untreated OSA may be associated with more severe respiratory failure in hospitalized patients with COVID-19. METHODS: Patients hospitalized in the Pulmonology Department with confirmed COVID-19, University Hospital in Kraków, Poland, between September 2020 and April 2021 were enrolled. OSA screening questionnaires including Epworth Sleepiness Scale (ESS), STOP-BANG, Berlin questionaire (BQ), OSA-50, and No-SAS were completed. Polygraphy was performed after > 24 h without requirement for supplemental oxygen. RESULTS: Of 125 patients with median age of 61.0 years, 71% of whom were male. OSA was diagnosed in 103 patients (82%) and was categorized as mild, moderate, and severe in 41 (33%), 30 (24%), and 32 (26%), respectively. Advanced respiratory support was introduced in 85 patients (68%), and 8 (7%) patients eventually required intubation. Multivariable analysis revealed that increased risk of requirement for advanced respiratory support was associated with higher respiratory event index (OR 1.03, 95%CI 1.00 to 1.07), oxygen desaturation index (OR 1.05, 95%CI 1.02 to 1.10), and hypoxic burden (1.02 95% CI 1.00 to 1.03) and lower minimal SpO2 (OR 0.89, 95%CI 0.81 to 0.98), but not with results of OSA screening tools like BQ score (OR 0.66, 95%CI 0.38 to 1.16), STOP-BANG score (OR 0.73, 95%CI 0.51 to 1.01), NoSAS score (OR 1.01, 95%CI 0.87 to 1.18), or OSA50 score (OR 0.84, 95%CI 0.70 to 1.01). CONCLUSION: Previously undiagnosed OSA was common among hospitalized patients who survived the acute phase of COVID-19. The degree of OSA was associated with the severity of respiratory failure.

Serum IL-17 and TNFα as prognostic biomarkers in systemic sclerosis patients: a prospective study.

Recent reports have demonstrated that endothelial injury is critical in the pathogenesis of systemic sclerosis (SSc) and is associated with increased levels of circulating inflammatory biomarkers. This study aims to analyze the serum concentrations of selected cytokines and evaluate their relationship with SSc clinics and the long-term course of the disease. This study included 43 SSc patients and 24 matched healthy controls. In both groups, we measured serum levels of inflammatory cytokines related to the inflammatory response, such as tumor necrosis factor (TNF)α, interferon (IFN)γ, interleukin (IL)-4, IL-6, IL-10, and IL-17, and fibroblast activation protein (FAP). Additionally, in SSc patients, we evaluated the presence of four single nucleotide polymorphisms (SNPs) located in the promotor region of the TNFA gene, namely rs361525, rs1800629, rs1799964, and rs1799724, which might be related to increased TNFα concentrations. The main aim consisted of associating inflammatory cytokines with (1) clinical disease characteristics and (2) longitudinal observation of survival and cancer prevalence. SSc patients were characterized by a 17% increase in serum TNFα. There was no other difference in serum cytokines between the studied groups and diffuse vs. limited SSc patients. As expected, evaluated serum cytokines correlated with inflammatory biomarkers (e.g., IL-6 and C-reactive protein). Interestingly, patients with higher IL-17 had decreased left ventricle ejection fraction. During the median 5-year follow-up, we recorded four cases of neoplastic diseases (lung cancer in two cases, squamous cell carcinoma of unknown origin, and breast cancer with concomitant multiple myeloma) and nine deaths. The causes of death included lung cancer (n = 2), renal crisis (n = 1), multiple-organ failure (n = 1), and unknown reasons in five cases. Surprisingly, higher TNFα was associated with an increased cancer prevalence, while elevated IL-17 with death risk in the follow-up. Furthermore, the AG rs361525 genotype referred to higher TNFα levels than GG carriers. Both AG rs361525 and CT rs1799964 genotypes were associated with increased cancer risk. Higher serum concentrations of TNFα characterize the SSc patients, with the highest values associated with cancer. On the other hand, increased IL-17 in peripheral blood might predict poor SSc prognosis. Further research is needed to validate these findings.

Data quality and patient characteristics in European ANCA-associated vasculitis registries: data retrieval by federated querying.

OBJECTIVES: This study aims to describe the data structure and harmonisation process, explore data quality and define characteristics, treatment, and outcomes of patients across six federated antineutrophil cytoplasmic antibody-associated vasculitis (AAV) registries. METHODS: Through creation of the vasculitis-specific Findable, Accessible, Interoperable, Reusable, VASCulitis ontology, we harmonised the registries and enabled semantic interoperability. We assessed data quality across the domains of uniqueness, consistency, completeness and correctness. Aggregated data were retrieved using the semantic query language SPARQL Protocol and Resource Description Framework Query Language (SPARQL) and outcome rates were assessed through random effects meta-analysis. RESULTS: A total of 5282 cases of AAV were identified. Uniqueness and data-type consistency were 100% across all assessed variables. Completeness and correctness varied from 49%-100% to 60%-100%, respectively. There were 2754 (52.1%) cases classified as granulomatosis with polyangiitis (GPA), 1580 (29.9%) as microscopic polyangiitis and 937 (17.7%) as eosinophilic GPA. The pattern of organ involvement included: lung in 3281 (65.1%), ear-nose-throat in 2860 (56.7%) and kidney in 2534 (50.2%). Intravenous cyclophosphamide was used as remission induction therapy in 982 (50.7%), rituximab in 505 (17.7%) and pulsed intravenous glucocorticoid use was highly variable (11%-91%). Overall mortality and incidence rates of end-stage kidney disease were 28.8 (95% CI 19.7 to 42.2) and 24.8 (95% CI 19.7 to 31.1) per 1000 patient-years, respectively. CONCLUSIONS: In the largest reported AAV cohort-study, we federated patient registries using semantic web technologies and highlighted concerns about data quality. The comparison of patient characteristics, treatment and outcomes was hampered by heterogeneous recruitment settings.

GraphTar: applying word2vec and graph neural networks to miRNA target prediction.

BACKGROUND: MicroRNAs (miRNAs) are short, non-coding RNA molecules that regulate gene expression by binding to specific mRNAs, inhibiting their translation. They play a critical role in regulating various biological processes and are implicated in many diseases, including cardiovascular, oncological, gastrointestinal diseases, and viral infections. Computational methods that can identify potential miRNA-mRNA interactions from raw data use one-dimensional miRNA-mRNA duplex representations and simple sequence encoding techniques, which may limit their performance. RESULTS: We have developed GraphTar, a new target prediction method that uses a novel graph-based representation to reflect the spatial structure of the miRNA-mRNA duplex. Unlike existing approaches, we use the word2vec method to accurately encode RNA sequence information. In conjunction with the novel encoding method, we use a graph neural network classifier that can accurately predict miRNA-mRNA interactions based on graph representation learning. As part of a comparative study, we evaluate three different node embedding approaches within the GraphTar framework and compare them with other state-of-the-art target prediction methods. The results show that the proposed method achieves similar performance to the best methods in the field and outperforms them on one of the datasets. CONCLUSIONS: In this study, a novel miRNA target prediction approach called GraphTar is introduced. Results show that GraphTar is as effective as existing methods and even outperforms them in some cases, opening new avenues for further research. However, the expansion of available datasets is critical for advancing the field towards real-world applications.

Association between clinical features and course of systemic sclerosis and serum interleukin-8, vascular endothelial growth factor, basic fibroblast growth factor, and interferon alpha.

BACKGROUND: Certain mediators, such as soluble growth factors and cytokines, among others, are implicated in the immunopathogenesis of systemic sclerosis (SSc). OBJECTIVES: This study aimed to examine the association between serum levels of vascular endothelial growth factor (VEGF), interleukin-8 (IL-8), interferon alpha (IFN-α), and basic fibroblast growth factor (bFGF) and the clinical presentation and course of SSc. MATERIAL AND METHODS: This longitudinal, observational study included 43 patients with SSc and 24 healthy subjects. Serum concentrations of VEGF, IL-8, IFN-α, and bFGF were measured at baseline in patients previously treated for SSc. Medical history of patients was analyzed retrospectively at the time of cytokine measurement to infer clinical correlations, and during follow-up for a median of 5 years, assessing the incidence of death or cancer. RESULTS: The bFGF and IFN-α concentrations differed between SSc patients and controls (p < 0.01). In turn, organ involvement and SSc phenotypes did not impact studied cytokine concentrations, similar to systemic steroid and/or immunosuppressant use at enrollment. However, we have documented a positive correlation between the current oral steroid dose and serum levels of IL-8 and bFGF. Furthermore, patients with a VEGF level ≥95.7 pg/mL and IFN-α level ≥3.6 pg/mL required cyclophosphamide therapy more often, currently or in the past (approx. 3-fold and 4-fold, respectively). Substantially elevated VEGF and IFN-α concentrations at baseline were associated with higher cancer occurrence (n = 4) during follow-up, while elevated circulating IL-8 level was associated with an increased risk of death (n = 9). CONCLUSIONS: The SSc group was characterized by higher serum concentrations of bFGF and IFN-α compared to healthy controls. Patients treated with cyclophosphamide or receiving higher systemic steroid doses, thus suffering from a more severe disease type, had increased cytokine levels. Elevated circulating IFN-α and VEGF levels might be correlated with cancer, whereas raised IL-8 levels may be associated with an increased risk of death. However, further research is needed to verify our findings.

Comprehensive Analysis of Circular RNAs in Endothelial Cells.

Non-coding RNAs constitute a heterogeneous group of molecules that lack the ability to encode proteins but retain the potential ability to influence cellular processes through a regulatory mechanism. Of these proteins, microRNAs, long non-coding RNAs, and more recently, circular RNAs have been the most extensively described. However, it is not entirely clear how these molecules interact with each other. For circular RNAs, the basics of their biogenesis and properties are also lacking. Therefore, in this study we performed a comprehensive analysis of circular RNAs in relation to endothelial cells. We identified the pool of circular RNAs present in the endothelium and showed their spectrum and expression across the genome. Using different computational strategies, we proposed approaches to search for potentially functional molecules. In addition, using data from an in vitro model that mimics conditions in the endothelium of an aortic aneurysm, we demonstrated altered expression levels of circRNAs mediated by microRNAs.

Decreased protein C activity, lower ADAMTS13 antigen and free protein S levels accompanied by unchanged thrombin generation potential in hospitalized COVID-19 patients.

INTRODUCTION: COVID-19 is associated with an increased thromboembolic risk. However, the mechanisms triggering clot formation in those patients remain unknown. PATIENTS AND METHODS: In 118 adult Caucasian severe but non-critically ill COVID-19 patients (median age 58 years; 73 % men) and 46 controls, we analyzed in vitro plasma thrombin generation profile (calibrated automated thrombogram [CAT assay]) and investigated thrombophilia-related factors, such as protein C and antithrombin activity, free protein S level, presence of antiphospholipid antibodies and factor V Leiden R506Q and prothrombin G20210A mutations. We also measured circulating von Willebrand factor (vWF) antigen and a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) antigen and activity. In patients, blood samples were collected on admission to the hospital before starting any therapy, including heparin. Finally, we examined the relationship between observed alterations and disease follow-up, such as thromboembolic complications. RESULTS: COVID-19 patients showed 17 % lower protein C activity, 22 % decreased free protein S levels, and a higher prevalence of positive results for IgM anticardiolipin antibodies. They also had 151 % increased vWF, and 27 % decreased ADAMTS13 antigens compared with controls (p < 0.001, all). On the contrary, thrombin generation potential was similar to controls. In the follow-up, pulmonary embolism (PE) occurred in thirteen (11 %) patients. They were characterized by a 55 % elevated D-dimer (p = 0.04) and 2.7-fold higher troponin I (p = 0.002) during hospitalization and 29 % shorter time to thrombin peak in CAT assay (p = 0.009) compared to patients without PE. CONCLUSIONS: In COVID-19, we documented prothrombotic abnormalities of peripheral blood. PE was characterized by more dynamic thrombin generation growth in CAT assay performed on admittance to the hospital.

Bronchial epithelial cell transcriptome shows endotype heterogeneity of asthma in patients with NSAID-exacerbated respiratory disease.

BACKGROUND: Nonsteroidal anti-inflammatory drugs-exacerbated respiratory disease (N-ERD) is currently classified as a type-2 (T2) immune-mediated disease characterized by asthma, chronic rhinosinusitis, and hypersensitivity to cyclooxygenase-1 inhibitors. OBJECTIVES: The aim of this study was to characterize immunological endotypes of N-ERD based on the gene expression profile in the bronchial epithelium. METHODS: mRNA transcriptome (mRNA-sequencing) was analyzed in bronchial brushings from patients with N-ERD (n = 22), those with nonsteroidal anti-inflammatory drug-tolerant asthma (NTA, n = 21), and control subjects (n = 11). Additionally, lipid and protein mediators were measured in bronchoalveolar lavage fluid (BALF). RESULTS: Initial analysis of the entire asthma group revealed 2 distinct gene expression signatures: "T2-high" with increased expression of T2-related genes (eg, CLCA1, CST1), and "proinflammatory" characterized by the expression of innate immunity (eg, FOSB, EGR3) and IL-17A response genes. These endotypes showed similar prevalence in N-ERD and NTA (eg, T2-high: 33% and 32%, respectively). T2-high asthma was characterized by increased expression of mast cell and eosinophil markers, goblet cell hyperplasia, and elevated LTE4 and PGD2 in BALF. Patients with a proinflammatory endotype showed mainly neutrophilic inflammation and increased innate immunity mediators in BALF. Furthermore, the proinflammatory signature was associated with a more severe course of asthma and marked airway obstruction. These signatures could be recreated in vitro by exposure of bronchial epithelial cells to IL-13 (T2-high) and IL-17A (proinflammatory). CONCLUSIONS: T2-high signature was found only in one-third of patients with N-ERD, which was similar to what was found in patients with NTA. The proinflammatory endotype, which also occurred in N-ERD, suggests a novel mechanism of severe disease developing on a non-T2 background.

Improvements in Quality Control and Library Preparation for Targeted Sequencing Allowed Detection of Potentially Pathogenic Alterations in Circulating Cell-Free DNA Derived from Plasma of Brain Tumor Patients.

Malignant gliomas are the most frequent primary brain tumors in adults. They are genetically heterogenous and invariably recur due to incomplete surgery and therapy resistance. Circulating tumor DNA (ctDNA) is a component of circulating cell-free DNA (ccfDNA) and represents genetic material that originates from the primary tumor or metastasis. Brain tumors are frequently located in the eloquent brain regions, which makes biopsy difficult or impossible due to severe postoperative complications. The analysis of ccfDNA from a patient's blood presents a plausible and noninvasive alternative. In this study, freshly frozen tumors and corresponding blood samples were collected from 84 brain tumor patients and analyzed by targeted next-generation sequencing (NGS). The cohort included 80 glioma patients, 2 metastatic cancer patients, and 2 primary CNS lymphoma (PCNSL) patients. We compared the pattern of genetic alterations in the tumor DNA (tDNA) with that of ccfDNA. The implemented technical improvements in quality control and library preparation allowed for the detection of ctDNA in 8 out of 84 patients, including 5 out of 80 glioma patients. In 32 out of 84 patients, we found potentially pathogenic genetic alterations in ccfDNA that were not detectable in tDNA. While sequencing ccfDNA from plasma has a low efficacy as a diagnostic tool for glioma patients, we concluded that further improvements in sample processing and library preparation can make liquid biopsy a valuable diagnostic tool for glioma patients.

Genetic Association between TNFA Polymorphisms (rs1799964 and rs361525) and Susceptibility to Cancer in Systemic Sclerosis.

Tumor necrosis factor (TNF)-α is a proinflammatory cytokine that plays an important role in the pathogenesis of autoimmune diseases. The aim of the study was to establish an association between TNF-α promoter variability and systemic sclerosis (SSc). The study included 43 SSc patients and 74 controls. Four single nucleotide polymorphisms (rs361525, rs1800629, rs1799724, and rs1799964) located at the promoter of the TNFA gene were genotyped using commercially available TaqMan allelic discrimination assays with real-time PCR. The rs1799724 allele was associated with an increased SSc susceptibility (p = 0.028). In turn, none of the polymorphisms studied were related to the clinical and laboratory parameters of SSc patients, except for a higher prevalence of anti-Ro52 antibodies in the AG rs1800629 genotype in comparison to GG carriers (p = 0.04). Three of four cancer patients had both CT rs1799964 and AG rs361525 genotypes; thus, both of them were related to the increased risk of cancer, as compared to the TT (p = 0.03) and GG carriers (p = 0.0003), respectively. The TNFA C rs1799724 variant is associated with an increased risk of SSc, while the CT rs1799964 and AG rs361525 genotypes might enhance cancer susceptibility in SSc patients, although large observational and experimental studies are needed to verify the above hypothesis.

Association of discharge policy with the length of hospital stay among patients with coronavirus disease 2019: a cross-sectional study.

AIM: To assess the association between discharge policy and hospital stay length, and to evaluate the factors related to duration of viral clearance among patients with coronavirus disease 2019 (COVID-19). METHODS: This cross-sectional study enrolled consecutive patients aged ≥18 years with SARS-CoV-2 infection confirmed by reverse transcription polymerase chain reaction test who were admitted to hospital. The participants were divided into the test-based (TB) policy group or symptom-based (SB) group depending on the policy valid at their hospital discharge. Multivariable analyses were performed to assess the factors related to the duration of hospital stay and viral clearance. RESULTS: The study involved 305 patients (66.6% men). The mean age was 60.9 (15.2) years. TB and SB policy groups consisted of 145 (47.5%) and 160 patients (52.5%), respectively. The TB group had significantly longer duration of hospital stay (21.0 vs 16.0, P=0.003). In multivariable analysis, SB policy was associated with significantly shorter hospital stay (ß-coefficient -5.87, 95% confidence interval [CI] -9.78 to -1.96, P=0.003). Longer viral clearance was associated with older age (ß-coefficient 0.33, 95% CI 0.15 to 0.51, P<0.001) and history of cough in the pre-hospital phase of the disease (5.96, 95% CI 0.64 to 11.29, P = 0.028). CONCLUSION: SB discharge policy is preferable in the context of limited resources during the COVID-19 pandemic.

MiR-191 as a Key Molecule in Aneurysmal Aortic Remodeling.

Abdominal aortic aneurysms (AAA) are a complex disease with an unclear pathomechanism. A positive family history is emphasized as a significant risk factor, and a nonspecific model of inheritance suggests participation of epigenetic regulation in the pathogenesis of this disease. Past studies have implicated microRNAs in the development of AAA; therefore in this project, we measured miR-191 levels in AAA patients and compared them with a control group. We found that miR-191 levels were significantly elevated in aneurysmal patients, although this did not correlate with the available clinical data. We then developed an in vitro model where, using cells with an endothelial phenotype, we determined the effect of miR-191 on the transcriptome using RNA sequencing. Subsequent pathway analysis established that some of the perturbations mediated by miR-191 can be explained by several processes which have long been observed and described in literature as accompanying the development of abdominal aortic aneurysms.

High-flow nasal oxygen therapy in the treatment of acute respiratory failure in severe COVID-19 pneumonia: a prospective observational study.

INTRODUCTION: A significant proportion of patients with COVID­19 present with a rapidly progressing severe acute respiratory failure. OBJECTIVES: We aimed to assess the efficacy of high­flow nasal oxygen (HFNO) therapy in severe acute respiratory failure in the course of COVID­19 in a noncritical care setting as well as to identify predictors of HFNO failure. PATIENTS AND METHODS: This prospective observational study was conducted between March and December 2020. We enrolled all consecutive patients hospitalized with confirmed SARS­CoV­2 infection in whom HFNO therapy was used. The primary outcome was death or endotracheal intubation within 30 days from admission. RESULTS: Of the 380 patients with COVID­19 hospitalized at our tertiary center, 116 individuals (30.5%) requiring HFNO due to severe pneumonia were analyzed. The primary outcome occurred in 54 patients (46.6%). The overall 30­day mortality rates were 30.2% (35 out of 116 patients) in the entire cohort and 64.7% (34 out of 51 patients) among individuals requiring endotracheal intubation. A multivariable analysis revealed that the ROX index (the ratio of oxygen saturation / fraction of inspired oxygen to respiratory rate) below 3.85 measured within the first 12 hours of therapy was related to increased mortality (hazard ratio, 5.86; 95% CI, 3.03-11.35) compared with the ROX index of 4.88 or higher. CONCLUSIONS: The results of our study suggest that nearly half of patients treated with HFNO due to severe COVID­19 pneumonia will require mechanical ventilation. The ROX index is a useful tool for predicting HFNO failure in this population.

A clinical profile and factors associated with severity of the disease among Polish patients hospitalized due to COVID-19 - an observational study.

INTRODUCTION: The coronavirus disease 2019 (COVID-19) is one of the greatest clinical challenges of the last decades. Clinical factors associated with severity of the disease remain unclear. The aim of the study was to characterize Polish patients hospitalized due to COVID-19 and to evaluate potential prognostic factors of severe course of the disease. MATERIAL AND METHODS: An observational study was conducted from March to July 2020 in the Pulmonology and Allergology Department of the University Hospital in Kraków, Poland. Consecutive patients with confirmed SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) infection were enrolled, and data about past medical history, signs and symptoms, laboratory results, imaging studies results, in-hospital management and outcomes was prospectively gathered. RESULTS: The study sample comprised 100 patients at the mean age of 59.2 (SD 16.1) years among whom 63 (63.0%) were male. Among them 10 (10.0%) died, 47 (47%) presented respiratory failure, 15 (15.0%) were transferred to the intensive care unit, 17 (17.0%) developed acute kidney injury, 7 (7.0%) had sepsis and 10 (10.0%) were diagnosed with pulmonary embolism. Multivariable analysis revealed age (OR 1.1; 95% CI 1.01-1.15), body mass index (BMI; OR 1.24; 95% CI 1.01-1.53), modified early warning score (MEWS; OR 3.95; 95% CI 1.48-12), the highest d-dimer value (OR 1.73; 95% CI 1.03-2.9) and lactate dehydrogenase (LDH; OR 1.16; 95% CI 1.03-1.3) to be associated with severe course of COVID-19. CONCLUSION: This observational study showed that almost half of hospitalized patients with COVID-19 developed respiratory failure in the course of the disease. Increasing age, BMI, MEWS, d-dimer value and LDH concentration were associated with the severity of COVID-19.

Time-dependent course of gastric ulcer healing and molecular markers profile modulated by increased gastric mucosal content of carbon monoxide released from its pharmacological donor.

BACKGROUND AND PURPOSE: Besides hydrogen sulfide (H2S) and nitric oxide (NO), carbon monoxide (CO) contributes to the maintenance of gastric mucosal integrity. We investigated increased CO bioavailability effects on time-dependent dynamics of gastric ulcer healing mediated by particular growth factors, anti-inflammatory and molecular pathways. EXPERIMENTAL APPROACH: Wistar rats with gastric ulcers induced by serosal acetic acid application (day 0) were treated i.g. throughout 3, 6 or 14 days with vehicle or CO-releasing tricarbonyldichlororuthenium (II) dimer (CORM-2, 2.5 mg/kg). Gross and microscopic alterations in gastric ulcer size and gastric blood flow (GBF) at ulcer margin were determined by planimetry, histology and laser flowmetry, respectively. Gastric mRNA/protein expressions of platelet derived growth factors (PDGFA-D), insulin-like growth factor (IGF-1), epidermal growth factor (EGF), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGFA) and their receptors, heme oxygenases (HMOX), nuclear factor (erythroid-derived 2)-like 2 (Nrf-2), cyclooxygenase (COX-2), hypoxia inducible factor (HIF)-1α, anti-inflammatory annexin-1 and transforming growth factor (TGF-ß1) were assessed by real-time PCR or Western blot. TGF-ß1-3 and IL-10 plasma concentration were measured using Luminex platform. Prostaglandin E2 content at ulcer margin was assessed by ELISA. KEY RESULTS: CORM-2 decreased ulcer area and increased GBF after 6 and 14 days of treatment comparing to vehicle. CO donor upregulated HGF, HGFr, VEGFR1, VEGFR2, TGF-ß1, annexin-1 and maintained increased IGF-1, PDGFC and EGF expression at various time-intervals of ulcer healing. TGF-ß3 and IL-10 plasma concentration were significantly increased after COMR-2 vs. vehicle. CONCLUSIONS: CO time-dependently accelerates gastric ulcer healing and raises GBF at ulcer margin by mechanism involving subsequent upregulation of anti-inflammatory, growth promoting and angiogenic factors response, not observed physiologically.

Biomarkers of inflammatory external root resorption as a result of traumatic dental injury to permanent teeth in children.

OBJECTIVE: External inflammatory root resorption (EIRR) is a common complication of traumatic dental injury (TDI) that can be detected radiologically. During EIRR, various proteins are released into gingival sulcus fluid (GCF). The aim of the study was to monitor the levels of selected proteins in GCF in children (8-16 years of age) in order to assess their utility in the early diagnosis of EIRR. DESIGN: Twenty five children who experienced TDI to permanent incisors with ended root development were enrolled. GCF was collected from injured and control teeth with paper strips within seven days after TDI and on three visits during six-month follow-up. Concentrations of IL-1α, IL-1ß, IL-6, IL-8, TNFα, RANKL and MMP-9 in GCF were measured using enzyme-linked immunosorbent assays. EIRR was confirmed by radiological imaging techniques. RESULTS: Of all analyzed proteins, only the levels of IL-1α, Il-1ß and TNFα in GCF from the injured teeth with resorption were higher than in GCF from control teeth on the visit during which the EIRR was diagnosed. In univariate logistic regression model, the concentration of IL-1α in GCF was found as the strongest risk factor for the occurrence of EIRR. CONCLUSIONS: The composition of GCF may be indicative of EIRR after TDI. The monitoring of selected biomarkers in GCF may help to detect EIRR at its early stage and might be useful in reducing radiological exposure in children after TDI. IL-1α can be considered as a potential marker of the EIRR in children after TDI to the permanent teeth.

Influence of TNF-α promoter variability on stage and grade in individuals with colorectal cancer.

Carcinogenesis is a multistep process in which inflammation plays an important role. Tumour necrosis factor a (TNF-α) is a cytokine that plays a major role in inflammation. Activity of the TNF cytokine family could influence progression of colorectal cancer (CRC). The aim of the study was to establish an association between TNF-α promoter variability and stage/grade in individuals with sporadic CRC. The study included 152 CRC patients and 107 healthy volunteers. Four single nucleotide polymorphisms (rs361525, rs1800629, rs1799724, and rs1799964) located at the promoter of TNFA gene were genotyped using commercially available TaqMan allelic discrimination assays by real-time PCR. CRC stage was described on the basis of preoperative imaging studies and postoperative histopathological report. The grade was described on the basis postoperative pathological examination of the specimen. In the case of rs361525, there was a statistically significant association with M-score (p = 0.0209). Rs361525 has significant association with tumour grade (p = 0.0260). We failed to demonstrate significant association between the other 3 SNPs and cancer grade. Rs361525 potentially could be under consideration when the survival rate and prognosis is assessed.

miR-29c-3p is an Effective Biomarker of Abdominal Aortic Aneurysm in Patients Undergoing Elective Surgery.

BACKGROUND: Abdominal aortic aneurysm (AAA) is usually asymptomatic and mostly diagnosed incidentally. Aortic dilatation progresses along with the wall weakening and eventually leads to a life-threatening rupture. Currently, there are no effective screening biomarkers for AAA. MicroRNAs, a class of small noncoding RNA molecules, offer great potential as biomarkers because of their stability in the bloodstream and expression profile specific for different diseases. OBJECTIVE: To assess the circulating miR-29c-3p as a potential biomarker of AAA and to elucidate the biological functions of miR-29c-3p in terms of pathogenesis of aneurysms. METHODS: Two groups of patients scheduled for elective surgery were studied: 52 patients with AAA, and 51 patients with peripheral artery disease who served as a control group (matched by age and gender). Serum miRNA was measured for circulating miR-29c-3p using quantitative real-time PCR. Functional tests of miR-29c-3p impact on the targeted transcripts were studied using its mimic or inhibitor and a cell culture of endothelial cells. RESULTS: Serum miR-29c-3p was significantly elevated in AAA patients as compared with controls (RQ=8.73) and correlated with the diameter of the aneurysm. No association between well-known risk factors and level of circulating miR-29c-3p was found using a logistic regression. In vitro study revealed that miR-29c-3p suppressed transcripts of ELN, COL4A1, PTEN and VEGFA. CONCLUSION: The results suggest that elevated miR-29c-3p is a potential serum biomarker for AAA. Causal involvement of miR-29c-3p in pathogenesis of the disease was found in human vascular endothelial cells, which extracellular matrix synthesis and integrity maintenance was inhibited.

Unbiased Profile of MicroRNA Expression in Ascending Aortic Aneurysm Tissue Appoints Molecular Pathways Contributing to the Pathology.

BACKGROUND: Complex etiopathogenesis of ascending aortic aneurysm suggests contribution of epigenetic mechanisms in its development. Several studies appointed microRNAs (miRs) as essential epigenetic factors in various human diseases; however, little is known about their role in ascending aortic aneurysm. Therefore, the aim of this study was to perform unbiased molecular screening of miRs expression in aneurysmal tissue and establish their functions on a transcriptional level. METHODS: Samples of ascending aortic tissue were obtained from 15 patients, and total RNA was isolated separately from aneurysmal and unaffected aortic tissue obtained from the same patient. Expression of the complete panel of human miRs was assessed by quantitative real-time polymerase chain reaction. Using bioinformatic tools, 13 genes were selected that were putatively regulated by overexpressed miRs. Expression level of transcripts were evaluated by quantitative real-time polymerase chain reaction and correlated with their targeting miRs. RESULTS: Overexpression of 10 miRs distinguished aneurysmal tissue from the unchanged one. These miRs were involved in cell senescence (miR-191-5p), maintenance of vascular integrity (miR-126-3p and miR-374-5p), nitric oxide-dependent vascular relaxation (miR-21-5p), smooth muscle differentiation, and contractility (miR-145- 3p, miR-29c-3p, miR-133a-3p, miR-186-5p, miR-143-3p, and miR-24-3p), and correlated with abundance of its miR targets. CONCLUSIONS: Altered expression of particular miRs selectively in the affected tissue indicate their role as factors that trigger pathways of aneurysmal transformation. Limited reparative properties due to overexpression of miR-191 may play a crucial role for aneurysm enlargement, whereas nitric oxide-dependent relaxation of vascular smooth muscle mediated by miR-21 offers an attractive explanation of the aneurysm's initiation, and is confirmed in experimental conditions.

Computed tomography characteristics of coronary artery atherosclerosis in subjects with lower extremity peripheral artery disease and no cardiac symptoms.

INTRODUCTION:  Computed tomography coronary angiography (CTCA) enables noninvasive evaluation of coronary artery atherosclerosis. However, its value to assess coronary artery disease (CAD) in subjects with lower­extremity peripheral artery disease (PAD) and no cardiac symptoms is unknown. Moreover, the relationship between coronary artery plaque characteristics and severity of peripheral atherosclerosis in this group of patients was not sufficiently elucidated. OBJECTIVES:  The aim of the study was to determine the value of CTCA to assess coronary artery atherosclerosis and to evaluate the relationship between coronary artery plaque characteristics and severity of peripheral atherosclerosis in subjects with lower­extremity PAD and no cardiac symptoms. PATIENTS AND METHODS:  Sixty­five individuals (45 men, 20 women, mean age, 62.5 ±7.6 years) with lower­extremity PAD and no cardiac symptoms underwent CTCA. RESULTS:  CTCA revealed CAD in 56 subjects. Twenty­two had obstructive CAD. The mean ankle-brachial index (ABI) was 0.64 ±0.16. Twenty­six individuals demonstrated abnormal carotid artery intima-media thickness (IMT). ABI lower than median, if compared with ABI equal of higher than median, was associated with a higher proportion of obstructive multivessel to single vessel CAD (8:4 vs. 1:9; P = 0.01) and higher number of coronary artery segments with mixed plaques (2.3 ±2.2 vs. 1.2 ±1.3; P = 0.02). Comparing patients with abnormal and normal IMT, the former demonstrated higher proportion of obstructive multivessel to single-vessel CAD (7:3 vs. 2:10; P = 0.01) and higher number of coronary artery segments with noncalcified (1.9 ±3.2 vs. 0.6 ±1.4; P = 0.04) and mixed plaques (2.3 ±2.1 vs. 1.3 ±1.7; P = 0.049). CONCLUSIONS:  CTCA may be effective to detect CAD in subjects with lower­extremity PAD and no cardiac symptoms. The low ABI and abnormal IMT are associated with more extensive CAD and higher burden of high­risk coronary artery plaques.