Search

Although, liver transplantation serves as the only curative treatment for patients with end- stage liver diseases, it is burdened with complications, which affect survival rates. In addition to clinical risk factors, contribution of recipient and donor genetic prognostic markers has been extensively studied in order to reduce the burden and improve the outcomes. Determination of single nucleotide polymorphisms (SNPs) is one of the most important tools in development of personalized transplant approach. To provide a better insight in recent developments, we review the studies published in the last three years that investigated an association of recipient or donor SNPs with most common issues in liver transplantation: Acute cellular rejection, development of new-onset diabetes mellitus and non-alcoholic fatty liver disease, hepatocellular carcinoma recurrence, and tacrolimus concentration variability. Reviewed studies confirmed previously established SNP prognostic factors, such as PNPLA3 rs738409 for non-alcoholic fatty liver disease development, or the role of CYP3A5 rs776746 in tacrolimus concentration variability. They also identified several novel SNPs, with a reasonably strong association, which have the potential to become useful predictors of post-transplant complications. However, as the studies were typically conducted in one center on relatively low-to-moderate number of patients, verification of the results in other centers is warranted to resolve these limitations. Furthermore, of 29 reviewed studies, 28 used gene candidate approach and only one implemented a genome wide association approach. Genome wide association multicentric studies are needed to facilitate the development of personalized transplant medicine.

Alcoholic liver cirrhosis (ALC) is the most common indication for liver transplantation (LT) in Croatia and presents a risk factor for the development of hepatocellular carcinoma (HCC). However, genetic susceptibility has not yet been systematically studied. We aimed to investigate the contribution of the risk polymorphisms PNPLA3 rs738409, EGF rs4444903, TM6SF2 rs58542926, MTHFR rs1801133, previously identified in other populations and, additionally, the contribution of Notch-related polymorphisms (NOTCH1 rs3124591, NOTCH3 rs1043996 and rs1044116, NOTCH4 rs422951). The study included 401 patients. The ALC group consisted of 260 LT candidates, 128 of whom had histopathologically confirmed HCC, and 132 of whom were without HCC. The control group included 141 patients without liver disease. Genotyping was performed by PCR using Taqman assays. The patients’ susceptibility to ALC was significantly associated with PNPLA3 rs738409, TM6SF2 rs58542926, and NOTCH3 rs1043996 polymorphisms. These polymorphisms remained significantly associated with ALC occurrence in a logistic regression model, even after additional model adjustment for sex and age. Cirrhotic patients with the PNPLA3 GG genotype demonstrated higher activity of ALT aminotransferases than patients with CC or CG genotypes. The susceptibility to the development of HCC in ALC was significantly associated with PNPLA3 rs738409 and EGF rs4444903 polymorphisms, and logistic regression confirmed these polymorphisms as independent predictors.

Aim: To investigate the association of FasL gene polymorphism (rs763110) with rheumatoid arthritis occurrence, disease activity, and tumor necrosis factor-α (TNF-α) plasma concentration in Croatian patients, and to conduct an updated meta-analysis. ----- Methods: This cross-sectional study enrolled 81 patients with rheumatoid arthritis and 94 control patients. After the assessment of the Disease Activity Score (DAS)-28, blood was taken for analysis. DNA was isolated from the whole blood to determine FasL polymorphism (rs763110) by polymerase chain reaction. Protein levels of TNF-α were determined with ELISA. After a detailed literature search, we conducted an updated meta-analysis using the Review Manager 5 software. ----- Results: Rheumatoid arthritis patients had significantly higher TNF-α concentration in plasma (1.65 [1.2-2.42] pg/mL) than controls (0.99 [0.77-1.35] pg/mL, P

Rheumatoid arthritis (RA) is an inflammatory joint disease that eventually leads to permanent bone and cartilage destruction. Fas has already been established as the regulator of inflammation in RA, but its role in bone formation under arthritic conditions is not completely defined. The aim of this study was to assess the effect of Fas inactivation on the bone damage during murine antigen-induced arthritis. Subchondral bone of wild-type (WT) and Fas-knockout (Fas-/-) mice was evaluated by histomorphometry and microcomputerized tomography. Proportions of synovial bone and cartilage progenitors were assessed by flow cytometry. Synovial bone and cartilage progenitors were purified by fluorescence-activated cell sorting and expression of Fas and Fas-induced apoptosis were analyzed in vitro. Results showed that Fas-/- mice developed attenuated arthritis characterized by preserved epiphyseal bone and cartilage. A proportion of the earliest CD200+ bone and cartilage progenitors was reduced in WT mice with arthritis and was unaltered in Fas-/- mice. During osteoblastic differentiation in vitro, CD200+ cells express the highest levels of Fas and are removed by Fas ligation. These results suggest that Fas-induced apoptosis of early CD200+ osteoprogenitor population represents potential mechanism underlying the impaired bone formation in arthritis, so their preservation may represent the bone-protective mechanism during arthritis.-Lazic Mosler, E., Lukac, N., Flegar, D., Fadljevic, M., Radanovic, I., Cvija, H., Kelava, T., Ivcevic, S., Sucur, A., Markotic, A., Katavic, V., Marusic, A., Grcevic, D., Kovacic, N. Fas receptor induces apoptosis of synovial bone and cartilage progenitor populations and promotes bone loss in antigen-induced arthritis.

Various reports suggest that adolescents seem to be obsessed with tanning. Existing evidence on attitudes demonstrate that young adults focus on improving appearance through a tan, with protective behaviors often being absent. The popularity of tanning, primarily among the young adult population, further promotes unsafe sun-related behaviors. The aim of this study was to determine the knowledge level of Mostar University students on the harmful effects of ultraviolet radiation (UVR), their attitudes and sun-related behaviors, and whether medical students are more likely to apply preventive measures against UVR in comparison with students from other faculties. The survey included a total of 140 undergraduate Mostar University students: 70 medical students and 70 non-healthcare-related faculty students. The data were collected by completing the personal design questionnaire created for this study. This study shows that Mostar University students have a high level of knowledge about UVR and skin cancer, but their behavior is not sufficiently preventive and appropriate to their knowledge. Medical students often use UV protection measures and employ more preventive behavior than that of other faculty students, but it is still at a low level of prevention. Findings concurred with existing evidence that the knowledge of associated risks from UVR was not sufficient to motivate safer sun-related practices.

25-Hydroxyvitamin D (25-OHD) may have a prognostic value in colorectal cancer (CRC) patients. However, as 25-OHD concentration is strongly impacted by surgery, it is uncertain what is the most reliable time-point for 25-OHD assessment, pre- or post-operative. Therefore, we examined 515 CRC patients (AJCC I-III) who underwent surgery. Blood samples were collected either pre- operatively (n = 286 ; median = 1 day before surgery) or post-operatively (n = 229 ; median = 8 days). Serum 25-OHD concentration was determined by liquid chromatography-tandem mass spectrometry. Association between 25-OHD and survival was tested in the whole cohort, followed by stratified analyses in pre- and post-operatively sampled. Median 25-OHD in the cohort was 36.7 nmol/L and median follow-up time was 5.9 years. There were no differences between pre- and post-operative cohort in age, sex, 25- OHD, AJCC stage, or localization of tumor. After adjustment, higher 25-OHD (>50 nmol/L) was associated with better overall survival only in post-operative (HR = 0.53 ; 95% CI: 0.33-0.84 ; P = 0.006), but not in pre- operative cohort (HR = 1.13 ; 95% CI: 0.77-1.65 ; P = 0.53). In conclusion, higher post-operative 25-OHD levels were associated with better survival outcome in CRC patients, while no such association was found for pre-operative levels. Time-point of blood collection should be addressed carefully in future research as it might affect the prognostic value of 25-OHD in CRC.

Supplemental Digital Content is available in the text, While increased serum concentrations of CXCL9/10 are associated with acute cellular rejection (ACR) occurrence, the association between CXCL9/10 single nucleotide polymorphisms (SNPs) and ACR after liver transplantation (LT) remains unknown. In the present case-control study, polymorphisms of CXCL9 (rs10336) and CXCL10 (rs3921) were determined by polymerase chain reaction in 215 liver transplant recipients. ACR was defined as biopsy proven within 6 months after LT. As selected SNPs were in 3’-UTR region, their possible association with protein synthesis was assessed by measuring the plasma concentration of CXCL9/10 in a cohort of 40 new transplant patients using ELISA. There was no association between CXCL9/10 genotypes and overall incidence of ACR. However, patients with CXCL9 genotype AA developed ACR earlier than patients with GG genotype (P = .003), with similar results for CXCL10 gene (CC vs GG; P = .005). There was no statistically significant difference in plasma concentrations of CXCL9/10 between the rejectors and the non-rejectors. Of note, patients with AA CXCL9 genotype had significantly higher CXCL9 plasma concentrations than patients with AG (P = .01) or GG genotype (P = .045). In conclusion, the SNPs of CXCL9 (rs10336) and CXCL10 (rs3921) are not associated with the incidence of ACR. However, patients with CXCL9 genotype AA developed ACR earlier and the same genotype was associated with greater plasma concentrations suggesting the involvement of CXCL9 mediated processes in ACR development.

Background and Aims: Despite the improvement and optimization of immunosuppressive protocols, acute cellular rejection (ACR) is still a frequent complication after liver transplantation. Recent studies showed that increased concentrations of chemokines CXCL9 and CXCL10 are associated with the ACR occurrence. The aim of this study was to examine the association of CXCL9 and CXCL10 single nucleotide polymorphisms with ACR after liver transplantation. Method: DNA was isolated from the whole blood of 215 patients transplanted due to alcoholic liver disease from 1/2009 to 3/2017 in University Hospital “Merkur”, Zagreb, Croatia. Polymorphisms of CXCL9 (rs10336) and CXCL10 (rs3921) were determined by polymerase chain reaction using commercially available TaqMan SNP assays. ACR was defined as biopsy proven (Banff score ≥3) within 6 months after liver transplantation. All patients received the same immunosuppressive protocol comprising of calcineurin inhibitors, steroids (discontinued after 3 months) and mycophenolate mofetil. Results: 59 patients with ACR and 156 patients without ACR were included into the study. There were no statistically significant differences in age, sex or MELD score at the time of transplant between rejection and non-rejection groups. Genotypes were in Hardy-Weinberg equilibrium (p > 0.05), with strong linkage disequilibrium (D’ = 0.99, r = 0986) between CXCL9 and CXCL10. In the rejection group 22 (37.3%) patients had GG, 25 (42.4%) had AG and 12 (20.3%) had AA genotype of the CXCL9 polymorphism, with similar genotype distribution observed in the non-rejection group, in which 54 (34.6%) patients had GG, 75 (48.1%) had AG and 27 (17.3%) had AA genotype. Lack of association between CXCL9 genotypes and incidence of ACR was found in codominant, dominant, recessive, overdominant or log- additive model (p > 0.05). Similar results were obtained for CXCL10 genotypes. On the other hand, we observed significant association between both CXCL9 and CXCL10 genotypes and the time of ACR occurrence. Patients with CXCL9 genotype AA developed ACR earlier (median 5 days (IQR 4.0–7.5)) than patients with the GG genotype (median 8 days (IQR 6.0–18.3)) (p = 0.003), with similar results for CXCL10 gene (CC vs GG ; p = 0.005). Conclusion: Single nucleotide polymorphisms of CXCL9 (rs10336) and CXCL10 (rs3921) are not associated with the incidence of acute cellular rejection after liver transplantation, but are associated with the time of ACR onset.

Rheumatoid arthritis (RA) is characterised by periarticular bone destruction leading to permanent disability. Some other forms of arthritis, such as arthritis in systemic lupus erythematosus (SLE), rarely produce bone lesions. Corresponding to that, inactivation of Fas produces a murine SLE-like phenotype, but alleviates bone loss during antigen-induced arthritis (AIA). Aim of this study was to identify cell populations differentially regulated in non- resorptive vs. resorptive arthritis by comparing the synovial and bone marrow cellular composition in Fas-deficient and wild-type mice with AIA. After obtaining approval from the Ethical Committee, mice were immunized with methylated(m)BSA in complete Freund’s adjuvant, followed by intra-articular injection of mBSA. Five weeks post-immunization, arthritis was assessed by histology and μCT. After collagenase digestion and labelling, cellular phenotype was determined by flow cytometry for the following markers: CD3, CD4, CD8, CD11b, CD29, CD31, CD44, CD45, CD90.1, CD106, CD115, CD166, CD117, B220, Gr-1, Sca-1, and TER119. Micro-CT confirmed pronounced decrease in epiphyseal subchondral trabecular bone volume in wt mice with AIA (22.31±3.74%), in comparison to control group (31.18±3.14%, P=0.002, t-test), and the decrease was absent in Fas −/− mice (29.57±3.07% in control vs. 27.98±4.08% in AIA, P=0.60, t-test). Proportions of B220+, CD3+ and CD11b+ cells were significantly increased (P=0.04, P=0.008, P=0.019, respectively, Kruskal-Wallis test), while the proportions of CD106+ and CD166+ non-haemopoietic cells were significantly decreased in the synovial compartment of wild-type mice with AIA (P=0.04, and P=0.05, respectively, Kruskal-Wallis test). In all mice with AIA, proportions of CD3+, CD11b+ and Gr-1+ cells were strongly negatively associated with bone volume (ρ0.60, P