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Sistemska skleroza (SSc) je sistemska avtoimunska revmatska bolezen, katere patogeneza ni popolnoma razjasnjena, tudi obvladovanje glavnih kliničnih znakov, kot sta vnetje in fibroza, s farmakoterapijo pogosto ni učinkovito. Z namenom boljšega razumevanja patogeneze in odkrivanja novih terapevtskih učinkovin smo proučevali oksidativni stres in migracijo monocitov pri bolnikih s SSc ter učinke adiponektina in resolvina D1 na vnetje in fibrozo. Metodološki pristop je zajemal delo na vzorcih bolnikov s SSc in zdravih oseb, celičnih kulturah in živalskem modelu. Pri bolnikih s SSc smo izmerili povišan indeks oksidativnega stresa (na osnovi povišanih serumskih reaktivnih kisikovih metabolitov in/ali antioksidantne kapacitete), več prelomov DNA v levkocitih ter oslabljene popravljalne mehanizme poškodb DNA. Na površini monocitov bolnikov s SSc smo dokazali povečano izražanje adhezijske molekule CD62L, katere izražanje je bilo spodbujeno s komponentami seruma bolnikov. Spremembe v oksidativnem stanju in migratornem fenotipu monocitov so pri bolnikih s SSc sovpadale s pljučno prizadetostjo, prisotnostjo avtoprotiteles in uporabo farmakoterapije. Na skeletnomišičnih celicah in skeletnomišičnih fibroblastih smo pokazali, da se adiponektin vpleta v vnetje in mišično regeneracijo z aktivacijo znotrajceličnih signalnih poti ter s spodbujanjem izločanja interlevkina-6 in sfingozin-1-fosfata, kot možne posledice povečane ceramidazne aktivnosti adiponektinskih receptorjev. AdipoRon, analog adiponektina, primerljivih učinkov ni izkazal. Resolvin D1 je na mišjem modelu peritonealne fibroze znižal delež polimorfonuklearnih celic v sistemskem krvnem obtoku, medtem ko vpliva na lokalno vnetje in fibrozo v trebušni steni ni imel. Negativno je vplival na izražanje hemerina, regulatorja vnetnih procesov, ki se veže na isti receptor kot resolvin E1. V doktorski disertaciji smo pokazali, kako obsežnejše stanje oksidativnega stresa in večji potencial migracije monocitov pri SSc sovpada s klinično sliko bolezni ter tako pripomogli k boljšemu razumevanju patogeneze. Obenem smo dokazali, da ima adiponektin učinke na mišično vnetje in regeneracijo, resolvin D1 pa potencial obvladovanja vnetja v fibrotičnih boleznih tudi z regulacijo izražanja ligandov resolvinskih receptorjev. Systemic sclerosis (SSc) is a systemic autoimmune rheumatic disease whose pathogenesis remains poorly understood, and pharmacotherapy for the major clinical manifestations - inflammation and fibrosis - is often ineffective. To better understand the pathogenesis and find new therapeutic options, we investigated oxidative stress and monocyte migration in SSc and effects of adiponectin and resolvin D1 on inflammation and fibrosis. Our methodological approach included work with samples from SSc patients, cell cultures, and animal models. SSc patients exhibited a higher oxidative stress index (based on increased serum reactive oxygen metabolites and/or antioxidant capacity), more DNA breaks in leukocytes, and impaired DNA damage repair mechanisms. We found increased surface expression of the adhesion molecule CD62L on SSc monocytes and showed that CD62L expression is stimulated by SSc serum components. Both the altered oxidative status and monocyte phenotype were consistent with lung involvement, autoantibodies, and pharmacotherapy in SSc patients. In skeletal muscle cells and skeletal muscle fibroblasts, adiponectin acted on inflammation and muscle regeneration by activating intracellular signalling pathways and stimulating secretion of interleukin-6 and sphingosine-1-phosphate, possibly due to increased ceramidase activity of adiponectin receptors. AdipoRon, an adiponectin analogue, did not show comparable effects. In a mouse model of peritoneal fibrosis, resolvin D1 decreased the proportion of polymorphonuclear cells in the systemic circulation, whereas locally in the abdominal wall it had no effect on inflammation and fibrosis. It also downregulated the expression of chemerin, an important regulator of inflammation that binds to the same receptor as resolvin E1. In our work, we show that oxidative stress and increased migratory potential of monocytes in SSc correspond to the clinical picture of the disease. We show that adiponectin has effects on muscle inflammation and regeneration and that resolvin D1 has the potential to promote resolution of inflammation by regulating the expression of other resolvin receptor ligands.

High-intensity exercise can elicit acute changes in the biochemical and physiological processes in the body of an athlete, including increased oxidative stress and inflammation. The purpose of this study was to explore the effect of acute moderate-intensity exercise on total antioxidant capacity (TAC) and serum levels of anti-inflammatory adiponectin (APN), and inflammatory markers in competitive cyclists. Ten male cyclists (age 15–26 years, body mass index 19.4–24.7 kg/m$^2$) participated in this study. Each subject performed the maximal oxygen uptake test (VO$_{2peak}$) and completed a 10-min cycling exercise at a workload of 50% of the peak of VO$_{2peak}$. Blood samples were collected on three different occasions: after an overnight fasting and at the exercise workloads of 50% and 100% VO$_{2peak}$. We measured APN, TAC, inflammatory markers as well as assessed nutrient and energy intake for each participant. Baseline concentration of serum APN (10.92µg/mL) significantly increased at 50% and at 100% VO$_{2peak}$. In addition, TAC also increased after acute exercise (0.079 vs 0.093 nmol/µL). The concentration of APN at 50% VO$_{2peak}$ positively correlated with the CRP (r = 0.640, p = 0.046) and negatively correlated with TNF-α (r = −0.696, p = 0.025). This test showed that short (10min) and medium-intensity (50% VO$_{2peak}$) exercise activity in trained athletes evoked beneficial antioxidant and anti-inflammatory responses. Importantly, this response correlates with the increase in APN levels thereby showing that highly trained individuals have beneficial responses originating from adipose tissue. Our observations show that a short training at moderate activity can be an important preservative strategy during the recovery training period.

Neonatalna hipoksično-ishemična encefalopatija (HIE) je posledica obporodnega pomanjkanja kisika v osrednjem živčevju ter je eden najbolj pogostih vzrokov za trajno nevrološko okvaro otrok. Pri nekaterih otrocih se kasneje razvije cerebralna paraliza (CP). Pri HIE pride do povečanega nastanka reaktivnih kisikovih spojin in aktivacije vnetnih procesov. Genetska variabilnost v teh poteh bi lahko vplivala na nastanek CP po HIE. V raziskavi smo se osredotočili na pogoste polimorfizme v ključnih genih, ki sodelujejo pri antioksidativni obrambi in pri vnetnem odgovoru po HIE. Vključili smo 90 otrok z zmerno in hudo HIE. S kompetitivnim alelno-specifičnim PCR smo določili polimorfizme v izbranih genih za antioksidativne encime (SOD2 rs4880, CAT rs1001179, GPX1 rs1050450) in provnetne dejavnike (NLRP3 rs35829419, CARD8 rs2043211, IL1B rs1143623, IL1B rs16944, IL1B rs1071671, TNF rs1800629). Pri otrocih, ki so bili zdravljeni s TH in so po rojstvu opravili MR slikanje glave, je bila stopnja možganske okvare opredeljena po klasifikaciji Rutheford. Povezanost polimorfizmov z razvojem CP po HIE in z izidom na MR smo analizirali z uporabo standardnih statističnih testov. Povezave med izbranimi polimorfizmi in nastankom CP nismo dokazali. V skupini otrok, ki ni bila zdravljena s TH, je bil polimorfen alel NLRP3 rs35829419 nominalno značilno povezan z večjim tveganjem za nastanek CP (p = 0,034). V skupini, zdravljeni s TH, pa preučevani polimorfizmi niso bili povezani s pojavom CP. V skupini, zdravljeni s TH, je bil polimorfni alel CARD8 rs2043211 nominalno značilno povezan z normalno/blago možgansko poškodbo na MR glave (p = 0,040), IL1B rs1143623 C alel pa je bil nominalno povezan z zmerno/hudo možgansko poškodbo na MR (p = 0,040). Hipoteze, da so polimorfizmi v izbranih genih povezani s pojavom CP po HIE, nismo potrdili. Vpliv polimorfizmov na izid po HIE je bil do sedaj slabo poznan, zato rezultati predstavljajo izhodišče za nadaljnja raziskovanja na tem področju z večjim številom preiskovancev. Neonatal hypoxic-ischemic encephalopathy (HIE) results from peripartum or intrapartum brain tissue hypoxia and ischemia and is a common cause of long-term neurologic sequelae, including cerebral palsy (CP). Hypoxic-ischemic (HI) insult results in oxidative stress and activation of inflammatory processes. Genetic variability in these pathways could influence the development of CP after HIE. The aim of our study was to evaluate the association of the selected single nucleotide polymorphisms (SNPs) in the genes involved in response to oxidative stress and inflammation with the development of CP after HIE. Ninety children with moderate and severe HIE were included in the study. Genotyping using competitive allele-specific PCR was performed for polymorphisms in genes encoding antioxidant enzymes (SOD2 rs4880, CAT rs1001179, GPX1 rs1050450) and proinflammatory factors (NLRP3 rs35829419, CARD8 rs2043211, IL1B rs1143623, IL1B rs16944, IL1B rs1071676, TNF rs1800629) that could contribute to the development of CP after HIE. In children treated with TH where MRI was performed, it was classified according to the Rutheford classification. Standard statistical tests were used to evaluate the association of selected SNPs with the development of CP. No association between the selected SNPs and the development of CP was found. Polymorphic NLRP3 rs35829419 allele was nominally significantly associated with the development of CP in children who were not treated with TH (p = 0.034). None of the SNPs were associated with CP in children treated with TH. Among children treated with TH polymorphic allele CARD8 rs2043211 was nominally significantly associated with normal/mild MRI overall findings (p = 0.040) and IL1B rs1143623 C allele was nominally significantly associated with moderate/severe MRI overall findings (p = 0.040). The association of genetic variability with the outcome after HIE has so far not been well established, therefore the results of our study represent foundations for future research in larger studies.

Uvod: Problem slepote kot posledice glavkoma se povečuje zaradi staranja prebivalstva. Nastanek in napredovanje primarnega glavkoma z odprtim zakotjem (POAG) sta povezana z oksidativnim stresom, vnetjem in apoptozo ganglijskih celic mrežnice. Različni dejavniki lahko povzročijo večje nastajanje reaktivnih kisikovih spojin in sproščanje mediatorjev imunsko-vnetnega odziva. Matriksne metaloproteinaze (MMP) so encimi, ki sodelujejo pri preoblikovanju izvenceličnega matriksa v trabekulumu in ciliarniku ter na ta način uravnavajo očesni tlak (IOP). Edini dokazano učinkovit način zdravljenja glavkoma je znižanje IOP. Z zdravili pogosto ne dosežemo zadostnega znižanja IOP, pri nekaterih bolnikih pa kljub nizkemu IOP bolezen napreduje. Latanoprost je prvi prostaglandinski analog, ki ga evropske glavkomske smernice, skupaj s selektivno lasersko trabekuloplastiko, priporočajo kot prvi izbor za zdravljenja glavkoma. Zaenkrat ne poznamo kliničnih ali bioloških označevalcev, s katerimi bi lahko napovedali tveganje za nastanek POAG, njegov potek ter odgovor na določeno vrsto zdravljenja. Hipoteze: Genetska variabilnost lahko vpliva na potek POAG. Farmakogenetska variabilnost napoveduje odgovor na zdravljenje z latanoprostom in s selektivno lasersko trabekuloplastiko (SLT) pri bolnikih s POAG in očesno hipertenzijo (OHT). Z ugotavljanjem genetskih označevalcev lahko napovemo, katero zdravljenje bo pri posameznem bolniku učinkovitejše in povezano z manj neželenimi učinki. Metode: V raziskavo smo vključili 307 bolnikov s POAG ali z OHT, starejših od 40 let in 339 kontrol. V retrospektivnem delu smo pri bolnikih in kontrolah analizirali polimorfizme posameznih nukleotidov (SNP) v genih, ki so vključeni v obrambo pred oksidativnim stresom (GSTM1, GSTT1, GSTP1, SOD2, CAT in GPX1), vnetne poti (TNF, IL1B in IL6) ter delovanje encimov MMP (MMP2, MMP3, MMP9, MMP14). V prospektivni del raziskave smo vključili nezdravljene bolnike, ki smo jih naključno razvrstili za zdravljenje z latanoprostom ali s SLT ter ovrednotili odgovor na zdravljenje po 1,5 meseca. Pri vseh bolnikih smo analizirali SNP v genih, ki so vključeni v antioksidativne in vnetne poti ter za encime MMP. Pri bolnikih zdravljenih z latanoprostom smo analizirali tudi SNP v genih, ki sodelujejo pri farmakodinamiki in farmakokinetiki latanoprosta (CES1,CES1P1, BCHE , SLCO2A1, PTGFR). Introduction: The problem of blindness because of glaucoma is increasing due to the aging population. The onset and progression of primary open-angle glaucoma (POAG) is associated with oxidative stress, inflammation, and apoptosis of retinal ganglion cells. Various factors cause increased formation of reactive oxygen species and release of mediators involved in the immune-inflammatory response. Matrix metalloproteinases (MMP) are enzymes involved in the formation of the extracellular matrix in the trabecular meshwork and ciliary body and thus regulate intraocular pressure (IOP). The only proven effective treatment is reduction of IOP. Medication often does not achieve sufficient reduction of IOP, and in some patients the disease progresses despite low IOP. The first prostaglandin analogue latanoprost and selective laser trabeculoplasty (SLT), have been both recommended as the first-choice treatment for POAG by the European Glaucoma Society Guidelines. Currently, there are no clinical or biological markers that would help to predict the risk of POAG, its course or the response to a particular type of treatment. Hypotheses: Genetic variability may influence the course of POAG. Pharmacogenetic variability predicts response to latanoprost and selective laser trabeculoplasty in patients with POAG and ocular hypertension (OHT). By identifying genetic markers, we can predict which treatment will be more effective and associated with fewer side effects in an individual patient. Methods: A total of 307 patients with POAG or OHT older than 40 years and 339 controls were included in the study. In the retrospective part, we analyzed common single nucleotide polymorphisms (SNPs) in the genes involved in the pathways of oxidative stress (GSTM1, GSTT1, GSTP1, SOD2, CAT and GPX1), inflammation (TNF, IL1B and IL6) and in the activity of MMP (MMP2, MMP9, MMP14). The prospective part of the study included treatment-naive patients randomized to treatment with latanoprost or SLT, and evaluated the treatment response after 1.5 months. In these patients we also analyzed SNPs in the genes involved in the oxidative and inflammatory pathways, and in MMP genes. In addition, in the latanoprost treatment group, we analyzed SNPs involved in the pharmacodynamics and pharmacokinetics of latanoprost (CES1, CES1P1, BCHE , SLCO2A1, PTGFR). Results: Homozygotes for GSTM1 gene deletion had a lower risk of developing OHT, while homozygotes for MMP3 rs3025058 had a higher risk of developing OHT. There were more subjects with a higher degree of glaucoma impairment among carriers of at least one GPX1 polymorphic allele, while among carriers of at least one MMP3 polymorphic allele there were more subjects with a lower degree of impairment. The SOD2 rs4880, CAT rs1001179, and TNF 1800629 polymorphisms were associated with an absolute decrease in IOP, whereas carriers of at least one copy of GSTT1 had a lower relative decrease in IOP after latanoprost treatment. IL6 polymorphism was associated with a smaller absolute and relative decrease in IOP after SLT treatment. The clinical-pharmacogenetic model better predicted the effectiveness of IOP reduction (30% and more) in SLT treatment compared to the clinical model, whereas in latanoprost treatment both models had similar predictive value. Conclusion: SNPs in genes involved in oxidative and inflammatory pathways may be potential biological markers for predicting the risk and progression of POAG, as well as the response to latanoprost and SLT treatment. A clinical-pharmacogenetic prediction model could be helpful in selecting a more effective treatment.

Oxidative stress and neuroinflammation are involved in the pathogenesis and progression of glaucoma. Our aim was to evaluate the impact of selected single-nucleotide polymorphisms in inflammation and oxidative stress genes on the risk of glaucoma, the patients’ clinical characteristics and the glaucoma phenotype. In total, 307 patients with primary open-angle glaucoma or ocular hypertension were enrolled. The control group included 339 healthy Slovenian blood donors. DNA was isolated from peripheral blood. Genotyping was performed for SOD2 rs4880, CAT rs1001179, GPX1 rs1050450, GSTP1 rs1695, GSTM1 gene deletion, GSTT1 gene deletion, IL1B rs1143623, IL1B rs16944, IL6 rs1800795 and TNF rs1800629. We found a nominally significant association of GSTM1 gene deletion with decreased risk of ocular hypertension and a protective role of IL1B rs16944 and IL6 rs1800629 in the risk of glaucoma. The CT and TT genotypes of GPX1 rs1050450 were significantly associated with advanced disease, lower intraocular pressure and a larger vertical cup–disc ratio. In conclusion, genetic variability in IL1B and IL6 may be associated with glaucoma risk, while GPX and TNF may be associated with the glaucoma phenotype. In the future, improved knowledge of these pathways has the potential for new strategies and personalised treatment of glaucoma.

Background Inflammation and oxidative stress are recognized as important contributors to Parkinson’s disease pathogenesis. As such, genetic variability in these pathways could have a role in susceptibility for the disease as well as in the treatment outcome. Dopaminergic treatment is effective in management of motor symptoms, but poses a risk for motor and non-motor adverse events. Our aim was to evaluate the impact of selected single-nucleotide polymorphisms in genes involved in inflammation and oxidative stress on Parkinson’s disease susceptibility and the occurrence of adverse events of dopaminergic treatment. Methods In total, 224 patients were enrolled, and their demographic and clinical data on the disease course were collected. Furthermore, a control group of 146 healthy Slovenian blood donors were included for Parkinson’s disease’ risk evaluation. Peripheral blood was obtained for DNA isolation. Genotyping was performed for NLRP3 rs35829419, CARD8 rs2043211, IL1β rs16944, IL1β rs1143623, IL6 rs1800795, CAT rs1001179, CAT rs10836235, SOD2 rs4880, NOS1 rs2293054, NOS1 rs2682826, TNF-α rs1800629, and GPX1 rs1050450. Logistic regression was used for analysis of possible associations. Results We observed a nominally significant association of the IL1β rs1143623 C allele with the risk for Parkinson’s disease (OR = 0.59; 95%CI = 0.38–0.92, p = 0.021). CAT rs1001179 A allele was significantly associated with peripheral edema (OR = 0.32; 95%CI = 0.15–0.68; p = 0.003). Other associations observed were only nominally significant after adjustments: NOS1 rs2682826 A allele and excessive daytime sleepiness and sleep attacks (OR = 1.75; 95%CI = 1.00–3.06, p = 0.048), SOD2 rs4880 T allele and nausea/vomiting (OR = 0.49, 95%CI = 0.25–0.94; p = 0.031), IL1β rs1143623 C allele and orthostatic hypotension (OR = 0.57, 95%CI = 0.32–1.00, p = 0.050), and NOS1 rs2682826 A allele and impulse control disorders (OR = 2.59; 95%CI = 1.09–6.19; p = 0.032). We did not find any associations between selected polymorphisms and motor adverse events. Conclusions Apart from some nominally significant associations, one significant association between CAT genetic variability and peripheral edema was observed as well. Therefore, the results of our study suggest some links between genetic variability in inflammation- and oxidative stress-related pathways and non-motor adverse events of dopaminergic treatment. However, the investigated polymorphisms do not play a major role in the occurrence of the disease and the adverse events of dopaminergic treatment.