Background/Objective: Canakinumab (CAN), a monoclonal antibody targeting interleukin-1 beta, has demonstrated efficacy in various autoinflammatory diseases (AIDs), particularly in inadequate response to colchicine in familial Mediterranean fever (FMF). This study aimed to evaluate the indications, efficacy, and safety of CAN based on real-life experience from a tertiary rheumatology clinic.Methods: This single-center study included 54 patients treated with CAN between May 2020 and September 2024. Patients were grouped as MEFV-positive FMF (n=42), MEFV-negative FMF (n=7), non-FMF autoinflammatory diseases (n=2), and adult-onset Still's disease (AOSD; n=3). Demographic and clinical data, treatment indications, response patterns, laboratory parameters, and adverse events were analyzed.Results: CAN was initiated mainly due to adverse effects (40.5%) or inadequate response (42.8%) to anakinra and colchicine. The median duration of CAN therapy was 22 months. Among MEFV-positive FMF patients, 81% achieved a complete response and 19% partial response. CAN significantly reduced attack frequency and duration, and improved inflammatory markers (CRP, ESR, WBC, and neutrophil count). Proteinuria decreased in a statistically significant but clinically modest manner following CAN treatment. Only 1 patient experienced reversible cytopenia. Dose intervals were successfully prolonged in 54.8% of MEFV-positive patients without loss of efficacy.Conclusions: Canakinumab is an effective and well-tolerated IL-1 beta inhibitor in FMF and other AIDs, particularly in patients who are inadequately responsive or intolerant to colchicine and anakinra. Real-world experience supports its sustained efficacy and the feasibility of dose interval extension in selected cases.

Capecitabine is a key component of colorectal cancer (CRC) chemotherapy; however, acquired resistance markedly limits its efficacy. MicroRNAs (miRNAs) are central regulators of drug response, and the flavonoid fisetin has been reported to exert anti-tumor and chemosensitizing effects. Here, we investigated whether fisetin modulates the expression of chemoresistance-associated miRNAs in a capecitabine-resistant CRC cell model. Candidate miRNAs were prioritized from the publicly available dataset GSE30894 using differential expression analysis, which identified miR-21, miR-181a, miR-203, miR-101, and miR-381 as differentially expressed between sensitive and resistant colorectal cancer cells. These miRNAs were quantified by RT-qPCR in parental HT29 and capecitabine-resistant HT29 (CR/HT29) cells under basal conditions and after 24-h exposure to capecitabine (40 µM), fisetin (120 µM), or the combination. Nuclear morphology was assessed by DAPI staining. At baseline, CR/HT29 cells showed lower expression of miR-381, miR-21, miR-203, and miR-101 and a trend towards higher miR-181a compared with parental HT29 cells. Treatment with capecitabine, fisetin, or their combination differentially modulated these miRNAs in the two cell lines. In both parental and resistant cells, fisetin-containing regimens reduced miR-21, while in CR/HT29 cells, fisetin significantly decreased miR-181a expression compared with fisetin-treated parental cells. Fisetin and fisetin + capecitabine also altered miR-203 and miR-101 levels, with a more pronounced suppression in HT29 than in CR/HT29 cells. These changes were accompanied by marked morphological alterations in resistant cells. Fisetin modulates chemoresistance-associated miRNAs in colorectal cancer, particularly in capecitabine-resistant HT29 cells, suggesting a chemosensitizing adjuvant role and offering an in vitro basis for in vivo studies.

Hepatic injury is one of the most critical problems in major liver surgeries, trauma, sepsis or shock. The novel Elabela (ELA) peptide was shown to exert protective effects against cardiac and renal injury. We hypothesized that ELA could also have protective effects in hepatic ischemia–reperfusion (HI/R) injury and associated remote organ injury. Male (n = 37) and female (n = 37) Sprague–Dawley rats were used. Rats were divided into short-term and long-term HI/R injury groups. Each group was then divided into saline-treated, N-acetylcysteine-treated (NAC, 150 mg/kg) and ELA-treated (40 μg/kg) subgroups. Immediately before hepatic ischemia and during reperfusion, rats were subcutaneously injected with saline, NAC or ELA, while injections in long-term groups were continued twice a day for four days. Short-term and long-term sham-operation groups received saline injections. Hepatic blood flow was measured via laser Doppler flowmetry. Intracardiac blood was obtained for analyses of aminotransferase, alanine aminotransferase, bilirubin, urea, creatinine and interleukin (IL)-6. Caspase-3 and 8-hydroxy-2'-deoxyguanosine levels were determined and histopathological analyses (hematoxylin–eosin and alpha-smooth muscle actin (SMA) immunohistochemical staining) were performed in hepatic tissues. Levels of malondialdehyde, antioxidant glutathione, myeloperoxidase activity, luminol and lucigenin-enhanced chemiluminescence were measured in liver, lung, and kidney. Significant improvement in hepatic blood flow was observed in both short- and long-term ELA-treated groups. HI/R-induced elevations in reactive oxygen species in all the studied tissues were decreased by ELA, indicating its efficient radical scavenging function similar to NAC treatment. ELA treatment improved hepatic function tests and alleviated liver fibrosis, as detected by increased alpha-SMA-immunoreactivity. Serum IL-6 levels were increased by ELA treatment, suggesting its role in the activation of IL-6-dependent intracellular pathways which may contribute to hepatocyte proliferation and liver regeneration. Similar to the common use of NAC in hepatic surgery, Elabela appears to have a therapeutic potential in alleviating the consequences of hepatic postreperfusion injury.

PURPOSE:Tofacitinib, a small molecule pan-JAK inhibitor, targets key inflammatory pathways that play a pivotal role in the pathophysiology of uveitis. Its ability to inhibit multiple cytokine signaling pathways makes it a promising candidate for the treatment of ocular inflammation. This study aimed to investigate the effect of oral tofacitinib on peptide-derived S-antigen (PDSAg)-induced chronic experimental autoimmune uveitis (EAU) in rats. MATERIALS AND METHODS:Nineteen albino Wistar rats were divided into five groups. Groups 1-3 (5 rats each) were immunized with 15 micrograms PDSAg to induce EAU; Group 2 received tofacitinib (5 mg/kg) by gavage twice daily. Group 3 received saline in the same manner as Group 2. Group 4 (2 rats) was a healthy control group. Group 5 (2 rats) received only tofacitinib. Uveitis development and treatment efficacy were evaluated using clinical scoring based on the the signs of anterior segment inflammation and histological scoring based on the deterioration of the retinal architectural structure. RESULTS:Uveitis confirmed based on histological evidence was observed in all EAU groups (Groups 1-3) compared to the healthy control group (Group 4) (p < 0.001, Mann-Whitney U test). Tofacitinib significantly delayed the onset of uveitis in Group 2 when compared with Groups 1 and 3, which did not receive tofacitinib (p < 0.001, Mann-Whitney U test). Histological scores also showed a trend toward reduction (p = 0.393, Mann-Whitney U test). CONCLUSIONS:Oral tofacitinib delayed uveitis onset and reduced clinical and histological findings, suggesting its potential as an alternative treatment for uveitis.

Objective Bamboo spine is the most severe complication of axial spondyloarthritis (AxSpA). This study aims to address whether haplotypes of endoplasmic reticulum aminopeptidase (ERAP)1 and ERAP2 single nucleotide polymorphisms (SNPs), previously associated with AxSpA, are associated with the development of bamboo spine in an AxSpA cohort. Methods The study included 192 patients with AxSpA followed in MARS (MARmara Spondyloarthritis) clinic and fulfilling the modified New York criteria. ERAP1 and ERAP2 polymorphisms were determined by real-time PCR on genomic DNA using the TaqMan SNP Genotyping Assay for SNPs rs72773968, rs3734016, rs26653, rs26618, rs27895, rs2287987, rs30187, rs10050860, rs17482078, rs27044, rs2549782, rs2248374. The HLA-B genotypes were assessed using the LABTypeTM SSO method. Results Male gender (P<0.001) and HLA-B27 positivity (P<0.05) were associated with an increased risk of developing a bamboo spine, and peripheral arthritis was significantly less prevalent in AxSpA patients with a bamboo spine (P<0.05). Of the ten ERAP1 haplotypes, haplotype 3 was significantly more prevalent in AxSpA patients with bamboo spine adjusted for HLA-B27 status (P<0.05). Haplotype 3 and -6 were significantly more prevalent exclusively in HLA-B27 (+) patients (P<0.05). Allele frequencies and genotype distributions of single ERAP1 SNPs alone were not significantly different between AxSpA patients with and without bamboo spines. There was no association between bamboo spine development and ERAP2 SNPs. Conclusions Our results indicate that HLA-B27 positivity, ERAP1 haplotype 3 and -6, and being male confer a significant risk for developing bamboo spine. Whether the association of haplotype 3 and -6 with bamboo spine is confined only to our Turkish patient cohort may deserve attention in other ethnicities.

ObjectivesBeh & ccedil;et disease (BD) has heterogeneous presentations, mainly mucocutaneous, vascular, and ocular manifestations. The mechanisms associated with different phenotypes have not been clarified. We aimed to investigate the expression of innate and adaptive immunity-related cytokines in these 3 main BD phenotypes in active and untreated states and remission after treatment to be able to develop a cytokine-based treatment algorithm.MethodsSerum samples were isolated from 41 patients with newly diagnosed active BD (aBD), which consisted of 19 mucocutaneous aBD, 11 ocular aBD (o-aBD), and 11 vascular aBD patients, 35 patients in remission (rBD), and 9 healthy controls (HC). Serum levels of each cytokine were measured with sandwich enzyme-linked immunosorbent assay and analyzed as both raw measurements and corrected levels for each 1 million white blood cells.ResultsThe study included 41 aBD patients (female/male [F/M]: 9/32; median age, 29 years), 35 rBD patients (F/M: 9/26; median age, 29 years), and 9 HC (F/M: 3/6; median age, 28 years). The serum interferon gamma level was significantly higher in the aBD group than in the rBD (116 vs. 92 pg/mL, p = 0.022). The serum interleukin 35 (IL-35) level was significantly higher in the HC group compared with aBD and rBD (p = 0.05). IL-17-related cytokines were lower in o-aBD. With treatment, they increased in o-aBD but decreased in mucocutaneous aBD and vascular aBD patients.ConclusionThis study supports the involvement of both innate and TH1-predominated adaptive immune responses across all BD phenotypes. The IL-17 and TH17-related immune responses appear less prominent in ocular BD, which may explain the ineffectiveness of IL-17 blockade in treating ocular BD. These findings support the need for further studies using comprehensive gene expression analyses to develop targeted treatment strategies for BD phenotypes.

Radiographic progression in Ankylosing spondylitis (AS) is driven by mechanical strain. A well-balanced spine provides a favorable weight distribution across the entheses. Pelvic parameters are useful in assessing the shape of the spine. The present study aimed to prospectively investigate the predictive value of pelvic parameters for radiographic progression in AS. This non-interventional, observational, and prospective study enrolled AS patients fulfilling the modified New York criteria (mNY) currently under follow-up in the MARS (MARmara Spondyloarthritis) outpatient clinics. The primary objective was to investigate the relationship between the baseline pelvic parameters and radiographic progression in the spine. Two trained radiologists (EB, OB) independently assessed the modified Stoke Ankylosing Spondylitis Spine Score (mSASSS). An orthopedic surgeon (AHA) and a radiologist (EB) derived the pelvic parameters. Patients with no bridging or bamboo spine were included in the final analysis. Risk assessment for radiographic progression, defined as a two-unit increase in mSASSS or developing a new syndesmophyte every two years, was done using uni- and multivariate logistic regression analyses. Radiographs of 69 AS patients were analyzed. The median (IQR 25–75) prospective follow-up was 47.7 (34.6–52.8) months. Only 33.3

Background and Objectives: The shape of the spine is critical in maintaining posture on the vertical axis, and the morphology of the pelvis affects the distribution and dynamics of mechanical stress along the spine. Pelvis and spine angulations, called spinopelvic parameters, vary individually, hence, the site of mechanical load in individuals. Particular types of spino-pelvic parameters are associated with different types of degenerative diseases. However, whether baseline pelvic parameters affect the location of new bone formation and the degree of radiographic damage in patients with SpA is not known. This study aimed to prospectively investigate the effect of pelvic parameters on syndesmophyte formation and radiographic damage in the spine in a heterogeneous group of SpA patients. Methods: Radiographic SpA (r-SpA) patients currently under follow-up in the MARS (MARmara Spondyloarthritis) outpatient clinic were included. Two readers independently scored the radiographs according to the modified Stoke Ankylosing Spondylitis Spine Score (mSASSS), and two readers calculated the spinopelvic parameters. The average scores were used. The relationship between pelvic parameters and radiographic progression was investigated using logistic regression analyses adjusted for potential confounders. Interactions were tested. Results: Sixty-nine patients completed the prospective follow-up period, with a median (IQR 25-75) follow-up period of 47.7 (34.6-52.8) months. Of these patients, 40 (58%) were male, and 39/68 (58.2%) were HLA*B27 positive, 38 (55.1%) were under treatment with biological therapies during the follow-up period. Radiographic progression developed in 23 (33.3%) of 69 patients. The mean (SD) mSASSS and the mean (SD) the number of syndesmophytes of these patients, respectively: at the time of enrolment were: 8.5 (4.6), 2.1 (1.8); after two years of follow-up were: 11.1 (4.28), 2.6 (1.9); at the end of the study were: 14.5 (4.7), 5 (2.7). Patients with radiographic progression statistically significantly had a higher baseline mSASSS (p<0.001), syndesmophyte count (p<0.001), PT measurement (p=0.037), and SS/PI (p=0.034) ratio than those without. No statistically significant difference existed between those with and without radiographic progression in gender, smoking, BMI, HLA*B27 positivity, and receiving biological treatment rates (Table 1). When PI, SS, and LL, were compared, PI was lower, and SS was higher in those with radiographic progression than those without. However, the difference between both parameters did not reach statistical significance. Nevertheless, the SS/PI ratio was statistically significantly higher in those with radiographic progression (in groups with and without radiographic progression, respectively; mean (SD): 0.78 (0.13) and 0.71 (0.15); p=0.034). Multivariate logistic regression analysis revealed that SS with an OR of 1.14 (95%CI: 1-1.31 p=0.043) and PI with an OR of 0.9 (95%CI: 0.82-0.996, p=0.041) were significant risk factors that are independent of the effects of previously reported risk factors or new bone formation. Conclusion: The SS/PI ratio is significantly higher in patients with radiographic progression. The baseline SS/PI ratio is correlated with the progression of mSASSS per unit time under follow-up, and the PI itself has a negative correlation with the progression in the lumbar spine. These correlations are more emphasized in the male sex and increase with age. In this model of new bone formation, information on HLA*B27, obesity, uveitis, smoking, and treatment had no additional contribution. REFERENCES: NIL. Acknowledgements: NIL. Disclosure of Interests: None declared.

Capecitabine is recommended as one of the first-line chemotherapy treatments for advanced or metastatic colorectal cancer. Researches have been conducted on capecitabine’s impact on the viability of human colon cancer cells and its potential to induce apoptosis. However, even in cases initially responsive to treatment, the development of acquired resistance significantly limits its efficacy. Challenges still exist in effectively treating patients with chemotherapy, and developing new cytotoxic drugs is hindered by drug resistance. Fisetin alters the cell cycle, inducing apoptosis, inhibiting cancer cell proliferation, and enhancing the therapeutic effectiveness of chemotherapy drugs. This work aims to create a plan for reversing capecitabine resistance. For this purpose, the role of capecitabine and/or fisetin combinations in cell proliferation and apoptosis has been determined in both wild-type and capecitabine-resistant HT29 cells (CR/HT29). We developed capecitabine-resistant cell line from wild-type HT29 cells. This study demonstrated the effects of capecitabine, fisetin, and their combinations on both resistant and wild-type cells through experiments including cell survival skills, cell proliferation, wound healing, colony formation, hoechst staining, and western blot analysis. We established capecitabine-resistant cell lines. P-gp expression increased in CR/HT29 cells. Capecitabine effects on a CR/HT29 cells less than wild-type HT29 cells. The combination of fisetin and capecitabine in cell proliferation caused greater reductions in wild-type HT29 cells than in capecitabine-resistant cells. Fisetin has also additive effects on the apoptotic pathway in CR/HT29 cells. This study provides new perspectives on the combination of capecitabine and/or flavonoid treatment in resistant cells.

Abstract Ankylosing Spondylitis is an autoimmune disease leading to inflammation in the joints and ligaments of the spine. ERAP1 is a major risk factor for AS and ERAP1 mutations may result in structural changes that alter the trimming efficiency, thereby altering the immune response. The underlying structural mechanisms of AS pathogenesis have not yet been fully elucidated. This study investigated ERAP1/ERAP2 allotypes using Molecular Dynamics in both monomeric and dimeric forms. ERAP1's domain IV has been found to be a favorable region for dimerization. Different allotype dimers exhibited different stability characteristics. Furthermore, the effects of allotypic variation were more pronounced in Hap2-/Hap8-coupled dimer structures and were more distinct in heterodimers. An analysis of the interchain region revealed that both H-bonding and electrostatic interactions between chains of Hap2–N392 heterodimer structures were lower than those between Hap2–Hap2 revealing that allotypic variations played a significant role in stabilizing and destabilizing dimer structures.

Objective: Ankylosing spondylitis (AS) is a chronic systemic inflammatory disease that leads to structural and functional impairments and reduced quality of life, with heterogeneous manifestations. The origin and possible role of extracellular vesicles represented by exosomes (EVexo) in the pathogenesis of AS were examined in this study. Materials and Methods: Extracellular vesicles (EVs) were isolated from serum from ten AS patients and ten healthy controls through Izon qEV2/35 nm columns. After assessing the isolate purity by bicinchoninic acid assay (BCA) and Enzyme-Linked ImmunoSorbent Assay (ELISA), the relationship between EVexo concentration and AS was tested by the BCA method. The EVexo surface markers were analyzed by flow cytometry (FC) to verify EVexo presence and reveal its origin. Results: In FC analysis, CD86+TSG101+ and CD3+TSG101+ exosome percentages of AS group were significantly higher than the control group (p<0.05). A significant difference was found between the AS and control groups in terms of CD3+IL17+ and CD3+IFNg+ and CD86+TNF alpha+ and CD86+IL12(p35)+ exosome percentages (p<0.01). Conclusion: The exosomes whose ratio increased in the AS process were derived from T cells expressing increased levels of IL-17A and IFNg in their membranes, and macrophages expressing increased levels of TNF alpha and IL-12(p35) in their membranes. The EVexo profile did not change according to the AS course.

Abstract Ankylosing spondylitis (AS) is a chronic inflammatory disorder affecting the axial skeleton and often associated with Human Leukocyte Antigen-B*27 (HLA-B*27) positivity. HLA-B*27 and its role in AS pathogenesis remain unclear despite the identification of multiple susceptibility alleles. As the most frequent subtype related to AS, HLA-B*27:05 differs from the non-associated HLA-B*27:09 subtype at a single position. This study focuses on the comparison of two subtypes in their binding to two arthritogenic peptides (ARGQPGVMG-DRASFIKNL) and a viral peptide (KK10) through 500 ns long molecular dynamic simulations. In the present study, it was found that peptide-MHC I complex stability and peptide presentation were similar when the peptides had similar C-terminal charges.