VIRŠKINIMO SISTEMOS TYRIMO INSTITUTAS (08)

Colorectal cancer (CRC) is one of the most common malignancies worldwide. Early and accurate diagnosis remains a clinical priority, yet current biopsy techniques are invasive, spatially limited, and may not capture the molecular heterogeneity of tumors. We evaluated the feasibility and diagnostic potential of electroporation-based biopsy (e-biopsy) as a minimally invasive technique for proteomic sampling of colorectal cancer tissues. We conducted a multicenter, multinational study involving 19 patients undergoing surgical resection for CRC. Paired tumor and adjacent normal tissues were sampled ex vivo using e-biopsy. Proteins extracted from each sample were analyzed via LC-MS/MS. Bioinformatics pipelines, including differential expression, PCA, and pathway analysis, were used to identify CRC-specific signatures. E-biopsy consistently retrieved more proteins from tumor tissues than from adjacent healthy tissues (mean: 1300 vs. 800). Of the 3246 proteins identified, 54% were significantly upregulated in tumor tissues. Notably, proteins such as DLAT, LETM1, RBBP4, PPIB, and BCAP31 emerged as potential CRC biomarkers. Functional analyses revealed dysregulation in RNA processing, immune response, and metabolic pathways, consistent with known CRC biology. The integrated workflow, spanning tissue collection, electroporation, protein isolation, and mass spectrometry analysis across four institutions in two countries, was successfully executed, demonstrating the feasibility of multi-institutional implementation of the e-biopsy protocol while preserving tissue integrity throughout. E-biopsy enables rapid, reproducible, and minimally invasive molecular sampling of CRC tissue. This study demonstrates its potential to complement standard histopathology, aid in early diagnosis, and support molecularly guided treatment strategies in colorectal oncology.

Introduction. The handbook provides recommendations that regulate the process of preparing the final master's theses (henceforth, FMT) in the integrated study programme Medicine. The document outlines the principles of work planning, research ethics, methodological requirements and scientific communication, thereby ensuring academic quality and the reliability of the research. [...].

Background and Objectives: Microscopic colitis (MC) encompasses two chronic inflammatory disorders of the large intestine: collagenous colitis (CC) and lymphocytic colitis (LC). Both conditions are characterised by chronic watery diarrhoea and substantially impaired quality of life. Diagnosis relies on colonoscopy with multiple biopsies, and no reliable non-invasive biomarker currently exists. This exploratory study aimed to investigate circulating fatty acid-binding protein 2 (FABP2), interleukin-10 (IL-10), and lipopolysaccharides (LPSs) as potential biomarkers for MC and to compare their profiles with those in ulcerative colitis (UC). Materials and Methods: Plasma samples were obtained from 45 patients with active CC, 16 patients with active LC, 52 healthy controls, 43 patients with active UC, and 43 patients with inactive UC. Concentrations of FABP2, IL-10, and LPS were measured by enzyme-linked immunosorbent assay (ELISA). Results: Plasma FABP2 concentrations differed significantly across groups (Kruskal–Wallis p = 0.008). CC patients exhibited the highest levels (median 1719.0 pg/mL, IQR 1364.0–2240.0) compared with active UC (median 1272.0 pg/mL, IQR 861.7–1727.5; p = 0.005) and inactive UC (median 1334.0 pg/mL, IQR 854.2–1702.0; p = 0.001), but did not differ significantly from controls (median 1364.5 pg/mL, IQR 982.6–2160.5; p = 0.076) or LC (median 1421.5 pg/mL, IQR 1207.0–2002.2; p = 0.171). IL-10 concentrations also differed across groups (Kruskal–Wallis p = 0.029 after removal of one extreme outlier in active UC). Active UC patients had significantly lower levels (median 2.6 pg/mL, IQR 1.4–4.6) than CC (median 4.0 pg/mL, IQR 3.0–7.2; p = 0.009) and controls (median 4.8 pg/mL, IQR 2.6–7.4; p = 0.005). LPS concentrations showed no overall differences across groups (Kruskal–Wallis p = 0.55), although CC patients had numerically higher levels (median 73.7 pg/mL, IQR 45.6–104.9) compared with controls (median 56.4 pg/mL, IQR 33.7–87.1; p = 0.124). No significant differences were observed between LC and other groups for any biomarker. Conclusions: In this exploratory study, plasma FABP2 and IL-10 showed limited diagnostic accuracy in differentiating CC from UC but failed to distinguish MC from healthy controls. LPS levels were not significantly different among study groups. None of the biomarkers reliably separated LC from other groups, possibly reflecting the small LC sample size. These preliminary findings suggest subtle differences in circulating biomarker profiles between CC and UC that warrant validation in larger cohorts.

Throughout Europe there are important differences in the structure and characteristics of Emergency Medical Services (EMS) and their response to out-of-hospital cardiac arrest (OHCA). The primary aim of EuReCa-THREE was to examine the epidemiology of cardiac arrest in Europe and explore the association between EMS response time and survival.

Background and aims: In primary biliary cholangitis (PBC), clinically significant portal hypertension (CSPH) may develop even in precirrhotic stages due to a presinusoidal component, potentially limiting the accuracy of Baveno VII (BVII) criteria for predicting esophageal varices (EV). We aimed to evaluate the utility of spleen stiffness measurement (SSM) in predicting EV and varices needing treatment (VNT) in patients with PBC. Method: This multicenter cross-sectional study included patients from 14 centers with PBC fulfilling at least one of the following criteria: Liver stiffness measurement (LSM) ≥8 kPa, splenomegaly, platelet count ≤150 × 109 /L, or ultrasonographic signs of cirrhosis or portal hypertension. LSM and SSM were assessed using transient elastography. Laboratory tests, ultrasound (±6 months from the SSM), and upper endoscopy (±12 m) were collected. Results: We included 126 patients, 87% women, with a median age of 64 years (IQR 58–71). Overall, 82% had a LSM ≥8 kPa, 48% thrombocytopenia, 49% splenomegaly, and 65% cirrhosis. Median LSM and SSM values were 13.6 kPa (IQR 9.1–21.3) and 36.6 kPa (IQR 25.2–48.6), respectively. EV were present in 39% of patients, of whom 28% had VNT. The prevalence of VNT was significantly higher in patients with LSM >15 kPa (24%) compared with those with LSM 8– 15 kPa (4%) or <8 kPa (0%; p = 0.002). For EV prediction, BVII criteria (LSM ≤ 15 kPa and platelet count ≥150 × 109 /L) showed moderate diagnostic accuracy (AUC 0.71), with a sensitivity (Se) of 85%, specificity (Sp) of 55%, and negative predictive value (NPV) of 85%. LSM alone demonstrated comparable performance (AUC 0.74, Se of 94%, Sp 48% and NPV of 93%) at a cut-off of 9.8 kPa. Platelet count also showed moderate accuracy (AUC 0.70, Se 55%, Sp 82%, NPV 74%) using a cut-off of 127.5 × 109 /L. SSM achieved the highest diagnostic accuracy for EV detection (AUC 0.79, Se 78%, Sp 74%, NPV 84%) at a cut-off of 37 kPa. Combining SSM ≥37 kPa with a platelet count <127.5 × 109 /L further improved diagnostic performance (AUC 0.81, Se 45%, Sp 88%, NPV 72%). For VNT prediction, SSM was the strongest predictor (AUC 0.85, Se 93%; Sp 65%; NPV 99%), at a cut-off of 39 kPa, outperforming BVII criteria, LSM, and platelet count (AUC 0.68, 0.74, and 0.77, respectively). Conclusion: SSM showed superior diagnostic accuracy for predicting esophageal varices and varices needing treatment in patients with PBC, improving BVII criteria. Its use may enable more accurate risk stratification, including in precirrhotic stages.

Background and aims: Prediction of hospitalizations is important in cirrhosis to ensure timely prevention. Microbial composition is altered in cirrhosis but functionality, which is the host-bacterial interface, needs further evaluation. Aim: define microbial pathways linked with 90-day hospitalization in a multi-national cirrhosis cohort. Method: Cirrhosis outpatients were recruited from 7 countries (USA, Türkiye, Korea, Denmark, Lithuania, Brazil & Mexico). Demographics, cirrhosis severity (MELD, complications, medications), diet, were recorded, stool collected & pts followed for 90 days for hospitalizations. Metagenomics & pathway analysis using HUMAnN were performed. PERMANOVA & regression analysis was performed using Maaslin2 for 90-day hospitalizations prediction. Results: 682 patients with cirrhosis (170 US, 90 Brazil, 99 Korea, 93 each Lithuania & Mexico, 84 Türkiye, & 53 Denmark) were enrolled. 60% were men, MELD 12 ± 5, albumin3.5 ± 0.70, Na 138 ± 7. Most common etiology was alcohol (35%) then viral (31%) & MASLD (17%). 56% were decompensated (35% on lactulose, 19% on rifaximin, 44% prior ascites, 19% prior variceal bleed, 31% on NS βblockers). Diet varied (most with yogurt & coffee). 90-day Hospitalizations: Seen in 169 (25%), most of which 133(79%) were liver-related. Country-wise rate differed with lowest in Brazil & Türkiye & highest in Lithuania & Korea. Bacterial pathway analysis: PERMANOVA adjusted for country showed differences between hospitalized versus not (p = 0.001). There were differences in pathway diversity (p = 0.004) but not richness. Maaslin2: Pathways ↓ in hospitalized: All had p < 0.01, carbohydrate storage and biosynthesis (sucrose synthesis II, glycogen degradation II), amino acid biosynthesis (L-ornithine & L-methionine synthesis), nucleotide & aromatic compound degradation Pathways ↑ in hospitalized fermentation, alternative substrate utilization, de novo nucleotide biosynthesis, cell wall recycling, lipid metabolism, and Enterobacteriaceae-associated functions (heterolactic fermentation, ethanolamine utilization, glycerol and hexitol fermentation, pyrimidine biosynthesis, & enterobacterial common antigen biosynthesis). Conclusion: In a multinational outpatient cirrhosis cohort, gut microbial functional pathways focused on urea cycle and bioenergetics intermediates were lower, while ammoniagenic and Enterobacterial pathways were higher in those who were hospitalized within 90 days. Microbial pathway profiling could improve risk prediction in cirrhosis beyond single center studies.

Background and aims: Metabolic dysfunction-associated steatotic liver disease (MASLD) is closely linked to obesity and type 2 diabetes, being now the second most common chronic liver disease in Europe, yet largely absent from policy agendas, exposing a major gap in Europe’s metabolic health response. This study aimed to provide the evidence on national policies and clinical practice guidelines (CPGs) for MASLD/metabolic dysfunction-associated steatohepatitis (MASH) in Europe. Method: MASLD/MASH policies (i.e., government strategies/action plans and CPGs) were reviewed in 25 European countries (in the European Union (EU) and the United Kingdom) and reported via a standardised data collection tool completed by a country lead. Each country’s data-collection tool was reviewed by a core study team of three researchers and cross-referenced with public documents in any language to ensure accuracy. Results: No country had a national MASLD/MASH strategy or action plan issued by a government body, though two (8.0%), Romania and Sweden, had sub-national strategies. Twelve (48.0%) had at least one national or subnational strategy or action plan in which MASLD/ MASH was mentioned (within alcohol, cancer, CVD, diabetes, healthy lifestyle/diet, liver disease, noncommunicable disease, obesity, or “other” strategies). Among the national or subnational strategies or action plans mentioning MASLD/MASH, obesity and diabetes (n = 9 out of 24 countries, 36.0% and n = 8, 32.0%, respectively) were the most frequent ones mentioning MASLD/MASH. Fifteen (60.0%) countries had national level MASLD/MASH CPGs. Regarding the mention of MASLD/MASH in national level clinical guidelines for dyslipidaemia, obesity, diabetes, alcohol, hypertension, CVD, end of stage liver disease (ESLD)/cirrhosis, liver cancer, liver transplantation, primary care, or “other,” 22 (88.0%) of the respondent countries had at least one clinical practice guideline mentioning MASLD/MASH. Obesity guidelines (n = 17, 68.0%), followed by guidelines for diabetes (n = 12, 48.0%), ESLD/cirrhosis (n = 10, 40.0%), liver transplantation (n = 6, 24.0%), and primary care (n = 5, 20.0%) most commonly included MASLD/MASH. Conclusion: MASLD remains insufficiently addressed in national policy, leaving health systems underprepared despite important diagnostic and therapeutic advances. Integrating MASLD/MASH into existing NCD frameworks and policies is urgently needed to address this public health threat.

Background and aims: Liver cancer is the 15th most common type of cancer in Lithuania (2022) and almost 50% of cases of hepatocellular carcinoma (HCC) were caused by the hepatitis C virus (HCV). The Lithuanian HCV screening programme started in 2022. HCV antibody testing by general practitioners (GP) was performed: 1) for people born 1945–1994 (once in a lifetime) and 2) for those who inject drugs (PWID) or HIV+ (annual testing). The aim of the study was to assess results of the first 39 months of the programme, evaluate different scenarios to achieve WHO 2030 targets and effect on HCC incidence. Method: GP invited people to perform a serum ant-HCV test. AntiHCV positive patients (pt) were referred to a gastroenterologist or infectologist for HCV RNA testing, and if result positive, direct-acting antiviral (DAA) was prescribed. Information about pt was obtained from National Health Insurance Fund. The Markov disease progression model elaborated CDA Foundation was used to assess HCV elimination progress. The data from the 2022–2023 screening were used as inputs. Three scenarios were developed: the ‘Base’ scenario - return to pre-screening program level in 2023 and 2 scenarios with different extents of treatment. Results: in 2022, about 1.8 million people born 1945–1994 lived in Lithuania. Between May 5, 2022 and July 31, 2025 from this population 1419059 (81%) were tested for HCV antibodies. Positive test were found in 1.3% of cases. In the risk group, 18695 PWID and HIV+ people were screened, of whom 29.4% were seropositive. Viremiawas detected in 52,8%. During 39-month period 3 times more HCV pt were treated with DAA than during the same period before starting of the program (8321 vs 2690). The Markov disease progression model to assess HCV elimination progress showed the following: Scenario 1: if the same number of pt are treated as before the screening, the WHO targets will not be reached. Scenario 2: treating 70% of infected patients will meet most but not all WHO targets. Scenario 3: by treating all infected patients by 2030, the WHO target will be met by saving 150 lives and preventing 90 new cases of decompensated cirrhosis and 120 cases of HCC. Conclusion: In country with moderate HCV seroprevalence, a population-based screening program can be feasible, screening 81% (more than 1,4 million) of the target population during the first 39 program months. In case of realization of the Scenario 3 we could expect to achieve WHO targets by 2030 and diminish HCC incidence in Lithuania.

Įvadas. Echinokokozė yra reta, lėtinė parazitinė liga, ku­rią Lietuvoje dažniausiai sukelia Echinococcus multilocu­laris (alveolinė echinokokozė, AE) ir Echinococcus gra­nulosus (cistinė echinokokozė, CE). Ši liga dažniausiai pažeidžia kepenis bei neretai diagnozuojama vėlyvose stadijose dėl ilgo latentinio periodo ir nespecifinių simp­tomų. Diagnozės patvirtinimui reikalingas kompleksinis ištyrimas, įskaitant radiologinius, serologinius ir histolo­ginius tyrimus, o gydymas dažniausiai susideda iš chi­rurginių intervencijų, taikomų kartu su antiparazitiniais preparatais. Tikslas. Pristatyti tretinio lygio universite­tinės ligoninės 20 metų patirtį, diagnozuojant ir gydant kepenų echinokokoze sergančius pacientus. Tyrimo medžiaga ir metodai. Į šį retrospektyvinį tyrimą įtraukti pacientų, sirgusių kepenų echinokokoze ir gydytų Lietuvos sveikatos mokslų ligoninės Kauno klinikų Chi­rurgijos klinikoje 2001–2021 metais, duomenys. Išanali­zavus pacientų duomenis, į galutinę analizę buvo įtraukti pacientai, kuriems patvirtinta kepenų echinokokozės dia­gnozė ir prieinami klinikiniai duomenys. Pacientai buvo atrinkti naudojant Tarptautinės statistinės ligų ir sveika­tos sutrikimų klasifikacijos dešimtojo leidimo Australi­jos modifikacijos (TLK-10-AM) kodus (B67.0-B67.9). Surinkti demografiniai duomenys, vaizdo ir serologiniai duomenys, antiparazitinio gydymo tipas, informacija apie chirurgines ar minimaliai invazines procedūras ir pooperacinės baigtys. Rezultatai. Į tyrimą įtraukta 100 pacientų (vid. amžius 55,8 m.), iš kurių 65% sudarė moterys. Dažniausiai nu­statytas sukėlėjas buvo E. multilocularis (47%), 33% – E. granulosus, o 20% atvejų sukėlėjas nebuvo iden­tifikuotas iki rūšies lygmens. Dauguma pacientų turėjo vieną židinį (44%), o 23% – keturis ar daugiau. Konser­vatyvusis gydymas taikytas 79% pacientų, dažniausiai skiriant albendazolą (66%). Chirurginis gydymas atliktas 75% pacientų – dažniausiai taikyta anatominė rezekcija (58,7%). Pooperacinės komplikacijos pasireiškė 24% pacientų. Mirties atvejis fiksuotas 1,3% pacientų. Išvados. Kompleksinis ištyrimas ir individualizuotas gy­dymas, taikytas tretinio lygio universitetinėje ligoninėje, lėmė palankias gydymo baigtis daugumai pacientų, pa­tvirtindamas centro lyderystę gydant kepenų echinoko­kozę Lietuvoje.