Abstract Background and Aims Malnutrition is a usually observed condition among patients on hemodialysis (HD) and is considered one of studiest indicators of mortality and morbidity. Malnutrition inflammation score (MIS) is associated with inflammation, nutritional status, quality of life, and 5-year prospective mortality. Also, phase angle (PhA) measured by bioelectrical impedance analysis (BIA) has been studied as indicator of nutritional status or muscle function in HD patients and lower PhA was associated with a greater risk of malnutrition. The aims of this study are to analyze the prevalence of malnutrition in hemodialysis (HD) patients in Croatia using MIS, and to assess the association of MIS and PhA with body mass composition, sociodemographic characteristics and laboratory parameters related to HD. Method 166 HD patients aged 68.5 (58.8-76) years, 55 women (33%) and 111 (67%) men with mean HD duration of 4 (2-6) years from 6 HD center were included in this study. For each study participant, data about body composition measured with BIA, laboratory, sociodemographic and clinical parameters were obtained. Also, body mass index (BMI) was calculated, and the Malnutrition Inflammation Score (MIS) used to assess nutritional status. Results Using an MIS cut-off point of 5 for malnutrition, 53% of patients were malnourished (MIS≥5). Therefore, those malnourished HD patients were significantly older (p = 0.001), with longer HD duration (p = 0.002), lower residual renal function (p = 0.02), waist circumference (p = 0.001), BMI (p < 0.001), muscle (p = 0.002) and fat mass (p = 0.001) in kg, phase angle PhA (p < 0.001), serum creatinine (p = 0.002), albumin (p = 0.008), phosphate (p = 0.01), and urate (p = 0.008) level but have significantly higher ferritin (p = 0.004), and mean cellular volume level (p = 0.02). Therefore, total MIS score significantly negatively correlated with HD duration (r = −0.247, p < 0.001), waist circumference (r = −0.323, p < 0.001),muscle mass (r = −0.236, p < 0.001), fat mass (r = −0.352, p < 0.001), serum creatinine (r = −0.271, p < 0.001), phosphor (r = −0.245, p < 0.001), urate (r = −0.344, p < 0.001), and TIBC (r = −0.376, p < 0.001), albumin (r = 0.241, p < 0.001), ferritin (r = 0.273, p < 0.001). In contrast, PhA significantly positively correlated with muscle mass (kg) (r = 0.311, p < 0.001), creatinine (r = 0.313, p < 0.001), phosphate (r = 0.208, p = 0.01), urate (r = 0.305, p = 0.01) and Total Iron Binding Capacity (TIBC) (r = 0.245, p < 0.001) while significantly negative correlation with ferritin (r = −0.182, p = 0.03) was found. Also, there was a significant negative correlation between PhA and MIS (r = −0.362, p < 0.001). A multivariate regression analysis showed that age (ß = 0.09, p = 0.005), waist circumference (ß = −0.09, p = 0.004), serum albumin (ß = −0.23, p = 0.04), and urate (ß = −0.01, p = 0.02), were predictive factors for malnutrition. Conclusion The prevalence of malnutrition in HD patients in Croatian HD population was high. These results indicate and highlight the urgent need for individualized and structural nutritional screening (using MIS score and PhA) and nutritional support in Croatian HD patients, especially for older HD patients and those with longer HD duration.

BACKGROUND:The effectiveness of the COVID-19 vaccine may differ in hemodialysis patients. The aim of this prospective multicenter study was to determine the degree of serological response to the SARS-CoV-2 vaccine in the population of dialysis patients and its association with later SARS-CoV-2 infections.METHODS:A blood sample was taken for the determination of COVID-19 serological status (IgG antibodies) in 706 dialysis patients 16 weeks after vaccination with the second dose (Pfizer-BioNTech).RESULTS:Only 314 (44.5%) hemodialyzed patients had a satisfactory response to the COVID-19 vaccine. Eighty-two patients (11.6%) had a borderline response, while 310 patients (43.9%) had an unsatisfactory (negative) post-vaccinal antibody titer. A longer dialysis vintage had an increased odds ratio (OR) of 1.01 for the occurrence of COVID-19 positivity after vaccination. In the group of subsequently positive patients, 28 patients (13.6%) died from complications of COVID-19. We have found differences in mean survival time between patients with and without appropriate responses to vaccination in favor of patients with a satisfactory serological response.CONCLUSIONS:The results showed that the dialysis population will not have the same serological response to the vaccine as the general population. The majority of dialysis patients did not develop a severe clinical picture or die at the time of positivity for COVID-19.

Data on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) reinfections in kidney transplant recipients (KTRs) are lacking. We conducted a retrospective observational study between March 2020 and May 2022 to determine the rate of SARS-CoV-2 reinfections and their outcome in 2837 KTRs. Reinfection was defined as positive SARS-CoV-2 reverse transcription–polymerase chain reaction after initial infection and proven eradication. The primary outcomes were the need for hospitalization during the reinfection and mortality. Sixty-two patients developed SARS-CoV-2 reinfection at the median of 11 mo after the first infection (2.2% of the total cohort and 7% of patients who experienced acute coronavirus disease 2019 [COVID-19]). Sixty-six percent of patients received at least 1 dose of the anti–SARS-CoV-2 vaccine before the reinfection. Median age was 51 y, 42% were female, and 42% were asymptomatic. Twenty-two patients required hospitalization during the reinfection for a median of 10 d. Three patients died, all from respiratory insufficiency. Two were fully vaccinated, and 1 received 1 dose of vaccine. The bivariate analysis identified 10 significant predictors for the hospitalization during the reinfection (Table 1). In a multivariate analysis, proteinuria (P = 0.03; OR = 4.99) and rehospitalization after primary acute COVID-19 (P = 0.004; OR = 11.09) significantly contributed to the model. Hospitalization after reinfection was necessary in 11 patients (17.7%). Three patients died after recovery from the reinfection. Two of them were not vaccinated. TABLE 1. - Predictors of hospitalization during the SARS-CoV-2 reinfection (bivariate logistic regression analysis) Predictors β P OR 95% CI History CMV 1.96 0.02 7.13 1.2-39.1 Steroid dose 0.21 0.04 1.324 1.01-1.52 Creatinine 0.01 0.03 1.01 1.001-1.02 CKD-EPI eGFR –0.04 0.003 0.96 0.94-0.98 Proteinuria (g/24 h) 1.88 0.01 6.57 1.54-27.92 Primary SARS-CoV-2 infection Hospitalization during acute covid 1.38 0.02 4.0 1.28-12.5 Pneumonia 1.42 0.01 4.13 1.35-12.67 Stop MMF/Aza 1.18 0.04 3.26 1.01-10.59 Rehospitalization—post-COVID 2.13 <0.001 8.4 2.38-29.6 SARS-CoV-2 reinfection Diarrhea 1.96 0.02 7.13 1.30-39.1 CKD-EPI, Chronic Kidney Disease Epidemiology Collaboration; CMV‚ cytomegalovirus; COVID‚ coronavirus disease; eGFR‚ estimated glomerular filtration rate; MMF‚ mycophenolate mofetil; SARS-CoV-2‚ severe acute respiratory syndrome coronavirus 2. The hospitalization during the initial infection was identified as a risk factor for reinfection.1 In our study, hospitalization during the primary COVID-19 episode was a risk factor for hospitalization during the reinfection. However, hospitalization during the post–COVID-19 was a much stronger risk factor and remained significant in multivariate analysis. This suggests additional clinical problems that may influence the overall immunosuppressive status and increase the risk of reinfection in KTRs. Proteinuria has already been reported as a significant risk factor for hospitalization during the reinfection,2 whereas better renal allograft function had a protective role in this and previous studies.3 In cancer patients, prior vaccination did not affect mortality from reinfection.4 In the study by Morris et al, 2.4% of organ transplant recipients developed reinfection. Two of them were fully vaccinated. However, their study did not include the Omicron variant.5 In our study, vaccination did not affect hospitalization during the reinfection or mortality from reinfection. The findings in this report are subject to several limitations. The observational nature of the study limits the ability to draw causal conclusions. We had no individual-level viral strain data. Vaccine efficacy was not determined. Strengths of the present study include its multicenter nature with a large number of patients and 100% nationwide coverage of several countries. The results are accurate as our patients have negative reverse transcription–polymerase chain reaction tests after the first SARS-CoV-2 infection, showing the true reinfection incidence. However, very high heterogeneity among patients and variables raises the possibility that the predictions for hospitalization and outcomes may not stand up with the accrual of greater numbers. In conclusion, COVID-19 reinfection can occur in KTRs and may be severe. Further work is urgently needed to better understand COVID-19 reinfections.

Background: This report describes data collected by the Croatian Registry of Renal Biopsies (CRRB) for the year 2019. Patients and methods: nine centers (82%) provided data for 255 native kidney biopsies. We assessed the anthropometric data, data on serum creatinine concentration (sCr), 24 h proteinuria, haematuria, serum albumin level, arterial hypertension, histological diagnosis, and complications after renal biopsy. Results: examined group consisted of 58% males, median age 58 y (18-80 y) and 42% women, median age 57 y (20-86 y). Males had a more impaired renal function at the time of renal biopsy, nephrotic syndrome, and hypertension. The most prevalent clinical presentation were urinary abnormalities (34.9%). Among all biopsy cases, primary glomerular diseases were the most often found histology group (41.5%), and IgA nephropathy was the most frequent diagnosis(47.1%). Among secondary glomerular diseases, pauci-immune glomerulonephritis (PIGN) was most often found (30.9%). The highest proteinuria was observed in minimal change disease and diabetic nephropathy (DN). The highest sCR values were found in membranoproliferative glomerulonephritis (MPGN) and necrotizing vasculitis. Patients with MPGN and DN had the highest blood pressure levels. Conclusion: CRRB provides important data on the epidemiology of biopsy-proven kidney diseases from the whole territory of Croatia

Cilj: Klinički bolnički centar (KBC) Osijek najmlađe je transplantacijsko središte za bubreg u Hrvatskoj. Prva presadba (TX, od engl. transplantation) u Osijeku učinjena je 2007. godine, u godini ulaska Hrvatske u Eurotransplant. Cilj istraživanja je prikazati rezultate transplantacije bubrega u KBC-u Osijek, od prve transplantacije učinjene 2007. godine do siječnja 2020. godine. Ispitanici i metode: U istraživanje su bili uključeni svi ispitanici (144) kojima je učinjena transplantacija bubrega u KBC-u Osijek. Iz medicinskih zapisa prikupljeni su demografski i medicinski podatci, vremenski podatci o dijalizi i TX-u te kreatininemiji kao mjeri funkcije presatka. Analiza preživljenja učinjena je Kaplan-Meierovim testom. Rezultati: Jednogodišnje preživljenje presatka cenzurirano za smrt s funkcionirajućim presatkom bilo je 93,8 %, a petogodišnje 88,9 %. Tijekom medijana praćenja od 7 godina (interkvartilni raspon, IQR, od engl. interquartile range, 4 – 9 godina), od min. 0 godina do maks. 13 godina, preživljenje pacijenata s funkcionirajućim presatkom bilo je 86,8 %. Medijan kreatininemije godinu dana nakon TX-a bio je 132 μmol/l (IQR 100 – 174 μmol/l ), od min. 48 μmol/l do maks. 492 μmol/l, a nakon pet godina 120 μmol/l (IQR 103 – 161 μmol/l), od min. 53 μmol/l do maks. 341 μmol/l. Zaključak: Rezultati bubrežnog TX-a u KBC-u Osijek bili su na razini europskih rezultata.

Introduction: The involvement of the high-mobility group box 1 protein (HMGB1) in various autoimmune and inflammatory diseases has been documented; however, the role of this proinflammatory molecule in children with diabetes type 1 (T1DM) has not been addressed. The aim of this case-control study is to compare the serum level of HMGB1 in children with newly diagnosed T1DM (group 1) and a control group composed of healthy children. Material and methods: This case-control study included 136 children: group 1 (n = 96) and a control group (n = 40). Measurements were taken from serum for the following: HMGB1, white blood cell count, C-reactive protein, glucose, haemoglobin A(1C) and beta-cell autoantibodies (GADA-65, IA-2, ICA). HMGB1 was determined using enzyme-linked immunosorbent assay on a Labsystems iEMS Reader MF analyser (Labsystems Diagnostics Oy, Helsinki, Finland). Results: The level (median and interquartile range) of HMGB1 was statistically higher (p < 0.001) in children with T1DM: 8.7 (5.0-9.8) mu g/l, in comparison with the control group: 1.0 (0.6-1.4) mu g/l. No correlation was found between HMGB1 and HbA1c in group 1, or between HMGBI and BMI. A statistically higher percentage of positive children for autoantibodies were present in group 1 compared to the control group (p <= 0.001). HMGB1 serum levels were also tested and the presence of autoantibodies, and none of those antibodies correlated with the level of HMGB1. Conclusions: The higher level of HMGB1 in children with T1DM, compared to the control group, indicates that this proinflammatory molecule is a good candidate marker of inflammation in children with T1DM.

Periodontal disease is a chronic multifactorial disease the worldwide incidence of which is higher than the incidence of caries and represents one of the leading problems in dental medicine. It is manifested by the loss of the attachment apparatus of the tooth and leads to the loss of teeth. Numerous studies have shown the association of periodontal disease and various chronic systemic diseases such as diabetes mellitus and cardiovascular disease. It is believed that low-grade level of chronic inflammation and release of bacterial toxins and inflammatory mediators in the bloodstream aggravate a chronic systemic disease. The purpose of our research was to investigate the possible association of periodontal disease and chronic kidney disease via the inflammatory cytokines path. In this cross-sectional study, we surveyed a total of 80 subjects divided into two groups. First group included subjects with chronic renal disease stages III and IV, and the second group included patients with chronic renal disease stage V that were on hemodialysis. We compared periodontal status, as well as serum levels of different cytokines, interleukin 6, interleukin 17A and tumor necrosis factor α between the two groups. The results showed no significant between-group differences in periodontal status, but interleukin 6 levels were significantly higher in the hemodialysis group of patients and were also associated with a poorer periodontal status.

Interleukin (IL)-10 is an anti-inflammatory cytokine, and a decrease in its secretion is associated with obesity, metabolic syndrome and type 2 diabetes. However, it has not been established whether the intensity of the immune response during diabetes-associated chronic inflammation affects the devel-opment and/or progression of type 2 diabetes and its microvascular complications. The aim of this study was to investigate the role of single nucleotide polymorphism (SNP)-1082G/A for IL-10 gene in development of diabetes type 2 and its complications. DNA was extracted from blood cells of 240 overweight/obese subjects for IL-10 genotyping. Based on the presence of diabetes type 2, patients were divided in two groups: experimental group of 144 patients with diabetes type 2 and control group of 96 age- and gender-matched subjects without diabetes. Compared to control group, diabetic group had higher levels of leukocytes (p=0.012), fibrinogen (p=0.049) and plasminogen activator inhibitor-1 (PAI-1) (p=0.009), and lower levels of albumin (p=0.001). There were no differences in the frequency of SNP-1082G/A for IL-10 gene between the two groups (p=0.654). When considering diabetes related traits in all subjects in relation to specific genotype, a group with homozygous (AA) genotype had higher values of the mean fasting glucose (p<0.000001), HbA1c (p<0.000001) and HOMA-IR (p=0.003632), while the mean HOMA-B value (p=0.000178) was lower when compared to the groups with GG and GA genotypes. There was no difference in development of diabetic nephropathy, retinopathy and polyneuropathy between the IL-10 polymorphism genotypes. In conclusion, obese diabetes type 2 patients had an increased inflammation activity compared to obese non-diabetic individuals. There was no association of the investigated polymorphisms and development of type 2 diabetes and its microvascular complications. However, diabetes related traits clearly depended on the presence of specific IL-10 genotype.

Background/Aims: Immune responses are involved in arterial hypertension. An observational cross-sectional case control study was conducted to estimate the association between Toll-like receptor 4 (TLR4) expression and interleukin (IL)-17A serum levels in patients with controlled and non-controlled hypertension. Methods: We have enrolled 105 non-complicated otherwise healthy hypertensive patients: 53 with well-controlled blood pressure and 52 non-controlled. TLR4 peripheral monocytes expression and serum IL-17A levels were determined by flow cytometry and ELISA, respectively. Results: Non-controlled patients exhibited higher TLR4 expression than well-controlled (25.60 vs. 21.99, P=0.011). TLR4 expression was lower in well-controlled patients who were prescribed beta blockers (18.9 vs. 22.6, P=0.005) and IL-17A concentration was higher in patients using diuretics in either group (1.41 vs. 2.01 pg/ml, P<0.001; well-controlled 1.3 vs. 1.8 pg/ml, P= 0.023; non-controlled 1.6 vs. 2.3 pg/ml, P=0.001). Correlation between IL-17A concentration and hypertension duration was observed in non-controlled patients (Spearman correlation coefficient . ρ=0.566, P<0.001) whereas in well-controlled patients a correlation was found between hypertension duration and TLR4 expression (ρ=0.322, P=0.020). Conclusions: Arterial hypertension stimulates the immune response regardless of blood pressure regulation status. Prolonged hypertension influences peripheral monocyte TLR4 expression and IL-17A serum levels. Anti-hypertensive drugs have different immunomodulatory effects: diuretics are associated with higher IL-17A concentration and beta-blockers with lower TLR4 expression.

4. hrvatski kongres o hipertenziji s međunarodnim sudjelovanjem 4 th Croatian Congress of Hypertension with International Participation Objective: Innate and adaptive immune responses have been involved in arterial hypertension.We conducted a study about Toll-like receptor 4 (TLR4) and interleukin-17A (IL-17A) in well regulated and unregulated hypertensive patients.One of the objectives in our study was to evaluate if the type of the used anti-hypertension therapy could influence TLR4 expression or IL-17A concentration.Design and Method: 105 hypertensive patients, without any other acute or chronic disease, have been involved, divided in two groups: 53 well regulated and 52 unregulated hypertensive patients.The patients had their IL-17A serum concentration determined with ELISA method and the TLR4 expression on peripheral monocytes applying flow cytometry. Results:The expression of TLR4 was much lower in the group of well regulated patients who were prescribed beta blockers (18.9 vs. 22.6, P=0.005) and the concentration of IL-17A was significantly higher in the patients with diuretics, in both groups (for all patients 1.41 pg/ml vs. 2.01 pg/ml P<0.001, well regulated patients: 1.3 pg/ml vs. 1.8 pg/ml, P= 0.023, unregulated patients: 1.6 pg/ml 2.3 pg/ml, P= 0.001).No significant differences were observed in TLR4 expression or IL-17A levels in the patients who received renin-angiotensin-aldosterone blockers (ACE inhibitors and AT1 receptor blockers) as part of their hypertension therapy.

Evidence indicates that that innate and adaptive immune responses are involved in development of arterial hypertension. Here, we conducted a case-control study to estimate the association between the expression of the TLR-4 receptor and IL-17A serum cytokine level in well regulated and unregulated hypertensive patients.

AimTo determine the prognostic value of baseline red blood cell distribution width (RDW) in diffuse large B cell lymphoma (DLBCL) patients.MethodsData from 81 DLBCL patients diagnosed from 2006 to 2013 at the University Hospital Center Osijek, Osijek, Croatia, were reviewed. We evaluated disease outcome, overall survival (OS) and event-free survival (EFS), and demographic, clinical and laboratory factors affecting outcome. Univariate analysis and Cox regression analysis were used.ResultsMedian age of patients was 64 years, 29 were men (35.8%). Higher RDW levels (%) were found in patients with advanced Ann Arbor clinical stage (14.94 ± 1.82 vs 13.55 ± 1.54, P = 0.001) and in those with poor response to therapy (14.94 ± 1.82 vs 13.55 ± 1.54, P = 0.001). Patients with RDW>15% (cut-off was calculated by receiver operating characteristics) had significantly worse OS (median [range], 33 months [20-46] vs 74 months [65-82], P < 0.001) and EFS (27 months [15-40] vs 68 months [59-77], P < 0.001). Cox regression analysis showed that RDW>15% was an independent prognostic factor for OS (HR 3.654, 95% CI 1.128-11.836) and EFS (HR 2.611, 95% CI 1.012-6-739).ConclusionHigh baseline RDW is an independent prognostic marker of poor outcome in patients with DLBCL. RDW could be an easily available and inexpensive marker for the risk stratification in patients with DLBCL.

Single nucleotide polymorphisms (SNP) of toll-like and NOD-like receptors have been associated with altered receptor activity and modified production of proinflammatory cytokines leading to a number of diseases. Our aim was to determine whether SNP of TLR2 (Arg753Gln), TLR4 (Asp299Gly, Thr399Ile), and NLRP3 (Q705K) influence susceptibility to juvenile spondyloarthrtis (jSpA) and juvenile idiopathic arthritis (JIA). After the DNA extraction, 26 patients with jSpA, 11 with oligoarticular, polyarticular, or systemic JIA, and 40 healthy controls were genotyped for Arg753Gln, Asp299Gly, Thr399Ile, and Q705K SNP using real-time PCR–SNP analysis. Statistically significant difference in genotype frequency for Thr399Ile SNP of TLR4 was observed in the jSpA (χ2 = 6.705, p = 0.035) and not in the JIA group (χ2 = 3005, p = 0.223). Regarding Asp299Gly SNP, no significant difference in genotype frequency was found; however, allele frequency was significant in both jSpA and JIA patients. No significant difference in genotype or allele frequency was observed for Arg735Gln and Q705K SNP. The399Ile polymorphism of TLR4 may be responsible for altered immune response to microbial infection in variant carriers and represent a mechanism of triggering overproduction of proinflammatory cytokines and long-term inflammation in jSpA. SNP of TLR2, NLRP3, and TLR4 (Asp299Gly) were not associated with jSpA or JIA.

OBJECTIVEThe aims of this study were to determine the HCV-RNA viral load, genotype distribution, risk factors and symptoms of HCVRNA positive viral load in HCV antibody-positive patients from north-eastern Croatia.MATERIALS AND METHODSFrom January 2009 to December 2011, 203 HCV antibody- positive patients (130 men and 73 women; median age 44.5 years) were analyzed for HCV-RNA by the COBAS TaqMan HCV test and genotyped by the Linear Array HCV Genotyping test (both from Roche). All patients completed a structured questionnaire about risk factors and symptoms.RESULTSThe HCV-RNA percentage was 61.1% and was similar for men and women. The HCV-RNA viral load increased with age: while 55% of 20-50 year old patients were HCV-RNA positive, 73% of patients >50 years were positive (p=0.021). Genotype 1 was the most prevalent genotype (79.8%), followed by 3 (12.9%), 4 (6.5%), and 2 (0.8%); genotypes 5 and 6 were not determined. Patients with genotype 1 (median, 50 years) were older than patients with 3 (median, 33.5 years) or 4 (median, 38 years). The blood transfusions performed in Croatian hospitals before 1993 was significantly associated with HCV-RNA positive viral load (p<0.05).CONCLUSIONThese data indicated an elevated prevalence of genotype 1 in elderly HCV-RNA positive patients and it may continue to rise. Using RNA-based detection in HCV positive-antibody patients would allow early detection of HCV in the acute stage of HCV disease and the increased risk of HCV genotyperelated treatment failure.

Chronic infection with hepatitis C virus (HCV) is caused by an inadequate immune response. Experimental data suggest that the impaired activation of Toll-like receptors (TLRs) 2 and 4 contributes to chronic infection. We assessed the distribution of three single-nucleotide polymorphisms (SNPs) in the TLR2 (Arg753Gln) and TLR4 (Asp299Gly/Thr399Ile) genes in individuals from north-east Croatia and their effect on the outcome of antiviral therapy. The study consisted of 60 chronically infected patients and 40 healthy subjects. TLR polymorphisms were determined by the PCR-based melting curve analysis. HCV genotyping was performed using the Linear Array Hepatitis C Virus Genotyping Test. Thirty-three patients were treated with standard interferon and ribavirin therapy, and their viral load was evaluated at weeks 28 and 53 after the beginning of therapy. The majority of chronic infections were caused by genotype 1 (77%), followed by genotypes 3 (15%) and 4 (7%). Patients with genotype 1 had higher viral loads than patients infected with other genotypes (P = 0.0428). Healthy individuals and patients with chronic infection had similar frequencies of TLR2-Arg753Gln and TLR4-Asp299Gly/Thr399Ile SNPs. Heterozygous and homozygous TLR4-Asp299Gly/Thr399Ile polymorphisms correlated with higher viral loads and delayed responses to antiviral therapy. We have provided the first evidence that TLR4 polymorphisms influence the success of antiviral therapy in our region. This suggests that therapeutic strategies should be adjusted not only according to HCV genotype but also to individual TLR polymorphism(s).

To investigate associations between the postoperative immune response and the levels of extracellular circulating DNA (cDNA), C-reactive protein (CRP), neutrophil/lymphocyte (N/L) ratio, and regulatory T (Treg) cells in the peripheral blood and their role as potential predictors of postoperative septic complications.

Introduction Single nucleotide polymorphisms of Toll like receptors modify cellular immune response and induce pro-inflammatory cytokine production and therefore could be associated with enthesitis related arthritis (ERA) and/or oligoarticular and polyarticular juvenile idiopathic arthritis (JIA). Objectives To determine whether polymorphisms of TLR2 and TLR4 influence susceptibility to ERA or JIA. Methods DNA was extracted from blood samples of 19 ERA patients, 10 patients with oligoarticular or polyarticular JIA and 40 healthy controls, all diagnosed according to ILAR criteria. Polymorphisms of the TLR2 (Arg753Gln) and TLR4 (Asp299Gly, Thr399Ile) were determined using real time and multiplex PCR. Results All JIA patients were carriers of wild type allele for all three polymorphisms. Regarding Arg753Gln polymorphism of TLR2, only one patient with ERA (5.56%) and 2 healthy controls (5%) were carriers of heterozygous allele. There were no homozygous mutants. All ERA patients had wild type allele for Asp299Gly polymorphism of TLR4. For Thr399Ile polymorphism of TLR4, 21.05% ERA patients were heterozygous (CT variant), and none of the ERA patients was homozygous (TT variant), (CC vs CT variant in ERA OR 3.7500, 95% CI 1.0498-13.3952, p 0.0419, CC vs TT variant OR 31.0000, 95% CI 1.7309-555.1867, p 0.0197). In group of healthy controls, TLR4 polymorphisms Asp299Gly and Thr399Ile were in linkage disequilibrium ; 2 controls were hetrozygous and 6 homozygous variant carriers for both polymorphisms, whereas linkage disequilibrium was not found among patient groups (CC vs CT in controls OR 16.0000, 95% CI 3.5905- 71.2994, p 0.0003, CC vs TT OR 5.3333, 95% CI 2.0099-14.1524, p 0.1944). Conclusion Polymorphisms of TLR2 and TLR4 are not associated with oligoarticular/polyarticular JIA. There was also no evidence that variants of TLR are major risk factors for ERA, however, lack of susceptibility should be confirmed on larger group of patients, since four out of 19 patients (21, 05%) were heterozygous. A study on larger cohort is currently ongoing.

Postoperative increase in inflammation biologic markers is associated with a nonspecific inflammatory response to a surgical injury. We investigated the kinetics of changes in serum concentrations of procalcitonin (PCT), C-reactive protein (CRP) and interleukin-6 (IL-6) after abdominal surgeries and we focused on the behaviour of those markers in the case of development of the systemic inflammatory response syndrome (SIRS). In the single centre we conducted a prospective observational study and we included patients admitted to the ICU after elective abdominal surgery. A total of 41 patients were included and 8 (19.5%) of them had clinical and laboratory signs of SIRS. Sepsis was confirmed in one of the patients, a 72-year old patient operated due to having an abdominal aortic aneurysm. Plasma concentrations of PCT, CRP and IL-6 were measured in all the patients before surgery and at the postoperative day 1 (POD1), postoperative day 2 (POD2) and postoperative day 3 (POD3). Systemic release of PCT, CRP and IL-6 was present in all the measured time points after the abdominal surgery. Median concentrations of IL-6 (100.4 pg/mL) and PCT (1, 17 pg/mL) production were measured highest at POD1 and the median of CRP (147 mg/L) was measured at highest POD2. A larger increase of all three measured markers was found in patients with SIRS compared to those without. IL-6 at POD1 and POD2 was a good predictor of SIRS (areas under curves were 0.71 and 0.765, respectively), showing the highest accuracy among investigated markers at those time points. CRP at POD3 was a good predictor of SIRS (AUC was 0.76). A cut-off of 95 mg/mL in the level of CRP at POD3 yielded a sensitivity of 87.5% and specificity of 66.7% in detecting SIRS. IL-6 and CRP were the best in detecting postoperative SIRS after abdominal surgery with the highest area under ROC curve. This study is showing that PCT is not a good marker of SIRS caused only by surgical injury without sepsis.