Roles of polymorphisms and expression of genes coding for chemokines CX3C ligand 1 and CXC ligand 16 and their receptors in the development and progression of multiple sclerosis in Serbia

Multipla skleroza je hroniĉna inflamatorna, autoimunska, demijelinizaciona i neurodegenerativna bolest centralnog nervnog sistema (CNS-a). Hemokini i njihovi receptori predstavljaju znaĉajne medijatore inflamacije koji uĉestvuju u patogenezi odreĊenih hroniĉnih inflamatornih i autoimunskih bolesti meĊu kojima je i multipla skleroza. Ciljni hemokini u ovoj studiji, CX3C ligand 1 (CX3CL1) i CXC ligand 16 (CXCL16), specifiĉni su po tome što postoje u dve forme - kao transmembranski adhezivni molekuli i kao solubilni hemoatraktanti koji nastaju nakon proteolitiĉkog seĉenja vanćelijskih hemokinskih domena njihovih transmembranskih formi. U toku inflamatornog odgovora, na membrani endotelnih vaskularnih ćelija eksprimirani su CX3CL1 i CXCL16, a na membrani leukocita receptori za CX3CL1 (CX3CR1) i CXCL16 (CXCR6), te ovi hemokini i njihovi receptori posreduju u prodiranju leukocita iz krvi u tkivo zahvaćeno inflamacijom, podsticanjem hemotaksije i adhezije leukocita za aktivirani endotel krvnog suda. Ova studija obuhvata genetsko-epidemiološku analizu polimorfizama zamena pojedinaĉnih nukleotida u kodirajućim regionima gena, koje rezultuju zamenama aminokiselina. To su polimorfizmi V249I i T280M u genu za CX3CR1, i I123T i A181V u genu za CXCL16. U prethodnim studijama je pokazano da ovi genski polimorfizmi menjaju funkcionalna svojstva CX3CR1 i CXCL16, kao i da su asocirani sa patogenezom odreĊenih hroniĉnih inflamatornih bolesti. Uzimajući to u obzir, ova studija je imala za cilj da po prvi put ispita asocijaciju navedenih polimorfizama u genima za CX3CR1 i CXCL16 sa nastankom i progresijom multiple skleroze. Primenom alel-specifiĉne PCR metode i PIRA PCR-RFLP metode detektovani su genotipovi polimorfizama V249I i T280M u genu za CX3CR1, kod zdravih kontrola i pacijenata sa multiplom sklerozom. UtvrĊeno je da haplotip I249T280 u genu za CX3CR1 ima znaĉajno veću uĉestalost kod pacijenata sa relapsno-remitentnom (RR) formom, u odnosu na pacijente sa sekundarno-progresivnom (SP) formom multiple skleroze, što znaĉi da ovaj haplotip ima protektivni efekat na progresiju RR u SP formu bolesti...Multiple sclerosis is a chronic inflammatory, autoimmune, demyelinating and neurodegenerative disease of the central nervous system (CNS). Chemokines and their receptors are important mediators of inflammation, which are involved in pathogenesis of certain chronic inflammatory and autoimmune diseases including multiple sclerosis. Chemokines of interest in this study, CX3C ligand 1 (CX3CL1) and CXC ligand 16 (CXCL16), are specific in that they can exist either as transmembrane adhesion molecules or soluble chemoattractants being generated by proteolytic cleavage of their transmembrane forms’ extracellular domains. During the inflammatory response, CX3CL1 and CXCL16 are expressed on the surface of vascular endothelium, while the leukocytes produce membrane receptors for CX3CL1 (CX3CR1) and CXCL16 (CXCR6). Therefore, these chemokines and their receptors mediate the infiltration of leukocytes from blood into the inflamed tissue areas, by stimulation of both chemotaxis and adhesion of leukocytes to the activated endothelium of blood vessels. This study is based on genetic epidemiological analysis of single nucleotide polymorphisms, which are located in the coding regions of genes and result in amino acids’ substitutions. These are V249I and T280M substitutions in the gene coding for CX3CR1, and I123T and A181V substitutions in the gene coding for CXCL16. In previous studies these polymorphisms have been associated with the functional properties of CX3CR1 and CXCL16 as well as the pathogenesis of certain chronic inflammatory diseases. Therefore, this study aimed to investigate the association of the polymorphisms in CX3CR1 and CXCL16 genes with the development and progression of multiple sclerosis. Using the allele-specific PCR and PIRA PCR-RFLP methods, genotypes of CX3CR1 V249I and T280M polymorphisms were detected in healthy controls and patients with multiple sclerosis. Following statistical analysis showed significantly higher frequency of CX3CR1 I249T280 haplotype in patients with relapsingremitting (RR) form, compared to patients with secondary-progressive (SP) form of multiple sclerosis, so this haplotype had a protective effect on progression of RR to SP form of the disease..

Roles of polymorphisms and expression of genes coding for chemokines CX3C ligand 1 and CXC ligand 16 and their receptors in the development and progression of multiple sclerosis in Serbia

Multipla skleroza je hroniĉna inflamatorna, autoimunska, demijelinizaciona i neurodegenerativna bolest centralnog nervnog sistema (CNS-a). Hemokini i njihovi receptori predstavljaju znaĉajne medijatore inflamacije koji uĉestvuju u patogenezi odreĊenih hroniĉnih inflamatornih i autoimunskih bolesti meĊu kojima je i multipla skleroza. Ciljni hemokini u ovoj studiji, CX3C ligand 1 (CX3CL1) i CXC ligand 16 (CXCL16), specifiĉni su po tome što postoje u dve forme - kao transmembranski adhezivni molekuli i kao solubilni hemoatraktanti koji nastaju nakon proteolitiĉkog seĉenja vanćelijskih hemokinskih domena njihovih transmembranskih formi. U toku inflamatornog odgovora, na membrani endotelnih vaskularnih ćelija eksprimirani su CX3CL1 i CXCL16, a na membrani leukocita receptori za CX3CL1 (CX3CR1) i CXCL16 (CXCR6), te ovi hemokini i njihovi receptori posreduju u prodiranju leukocita iz krvi u tkivo zahvaćeno inflamacijom, podsticanjem hemotaksije i adhezije leukocita za aktivirani endotel krvnog suda. Ova studija obuhvata genetsko-epidemiološku analizu polimorfizama zamena pojedinaĉnih nukleotida u kodirajućim regionima gena, koje rezultuju zamenama aminokiselina. To su polimorfizmi V249I i T280M u genu za CX3CR1, i I123T i A181V u genu za CXCL16. U prethodnim studijama je pokazano da ovi genski polimorfizmi menjaju funkcionalna svojstva CX3CR1 i CXCL16, kao i da su asocirani sa patogenezom odreĊenih hroniĉnih inflamatornih bolesti. Uzimajući to u obzir, ova studija je imala za cilj da po prvi put ispita asocijaciju navedenih polimorfizama u genima za CX3CR1 i CXCL16 sa nastankom i progresijom multiple skleroze. Primenom alel-specifiĉne PCR metode i PIRA PCR-RFLP metode detektovani su genotipovi polimorfizama V249I i T280M u genu za CX3CR1, kod zdravih kontrola i pacijenata sa multiplom sklerozom. UtvrĊeno je da haplotip I249T280 u genu za CX3CR1 ima znaĉajno veću uĉestalost kod pacijenata sa relapsno-remitentnom (RR) formom, u odnosu na pacijente sa sekundarno-progresivnom (SP) formom multiple skleroze, što znaĉi da ovaj haplotip ima protektivni efekat na progresiju RR u SP formu bolesti...Multiple sclerosis is a chronic inflammatory, autoimmune, demyelinating and neurodegenerative disease of the central nervous system (CNS). Chemokines and their receptors are important mediators of inflammation, which are involved in pathogenesis of certain chronic inflammatory and autoimmune diseases including multiple sclerosis. Chemokines of interest in this study, CX3C ligand 1 (CX3CL1) and CXC ligand 16 (CXCL16), are specific in that they can exist either as transmembrane adhesion molecules or soluble chemoattractants being generated by proteolytic cleavage of their transmembrane forms’ extracellular domains. During the inflammatory response, CX3CL1 and CXCL16 are expressed on the surface of vascular endothelium, while the leukocytes produce membrane receptors for CX3CL1 (CX3CR1) and CXCL16 (CXCR6). Therefore, these chemokines and their receptors mediate the infiltration of leukocytes from blood into the inflamed tissue areas, by stimulation of both chemotaxis and adhesion of leukocytes to the activated endothelium of blood vessels. This study is based on genetic epidemiological analysis of single nucleotide polymorphisms, which are located in the coding regions of genes and result in amino acids’ substitutions. These are V249I and T280M substitutions in the gene coding for CX3CR1, and I123T and A181V substitutions in the gene coding for CXCL16. In previous studies these polymorphisms have been associated with the functional properties of CX3CR1 and CXCL16 as well as the pathogenesis of certain chronic inflammatory diseases. Therefore, this study aimed to investigate the association of the polymorphisms in CX3CR1 and CXCL16 genes with the development and progression of multiple sclerosis. Using the allele-specific PCR and PIRA PCR-RFLP methods, genotypes of CX3CR1 V249I and T280M polymorphisms were detected in healthy controls and patients with multiple sclerosis. Following statistical analysis showed significantly higher frequency of CX3CR1 I249T280 haplotype in patients with relapsingremitting (RR) form, compared to patients with secondary-progressive (SP) form of multiple sclerosis, so this haplotype had a protective effect on progression of RR to SP form of the disease..

Gene expression of chemokines CX3CL1 and CXCL16 and their receptors, CX3CR1 and CXCR6, in peripheral blood mononuclear cells of patients with relapsing-remitting multiple sclerosis - a pilot study

Background/Aim. In vitro and in vivo studies show that CX3CL1 and CXCL16 chemokines and their specific receptors, CX3CR1 and CXCR6, respectively, mediate mechanism of neuroinflammation during the pathogenesis of multiple sclerosis (MS). The aim of this study was to investigate relative messenger ribonucleic acid (mRNA) levels of CX3CL1, CXCL16, CX3CR1 and CXCR6 in peripheral blood mononuclear cells, as potential molecular markers of relapsing-remitting (RR) MS. Methods. The study included 43 unrelated RR MS patients, 20 of them with clinically active disease (relapse) and 23 with clinically stable disease (remission), and 28 unrelated healthy subjects as controls. Real-time polymerase chain reactions (PCR) were performed using TaqMan? gene expression assays. Relative expression (mRNA) level of each target gene in each sample of peripheral blood mononuclear cells was calculated as the mean normalized expression. Results. The levels of CX3CR1 mRNA were significantly higher in clinically active RR MS patients compared to controls [fold change = 1.38, p (Mann-Whitney U test) = 0.009], and significantly lower in clinically stable vs active RR MS patients [fold change = - 1.43, p (t-test) = 0.03]. Stable RR MS patients had significantly higher CXCL16 mRNA levels than controls [fold change = 1.33, p (Mann-Whitney U test) = 0.006]. A trend of increased CXCR6 gene expression was found in active RR MS patients compared to controls [fold change = 1.23, p (Mann-Whitney U test) = 0.08]. In either active or stable RR MS patients there were no significant correlations of the clinical parameters with expression levels of the target genes. Conclusion. The current results show that increased CX3CR1 mRNA levels in peripheral blood mononuclear cells could represent a proinflammatory molecular marker of clinically active RR MS

HAp:Co as tunable VIS-NIR reflective pigment

Radar-absorbent materials, used in stealth technology for defense aircrafts, vehicles, satellites, etc. from radar detection, are commonly based on graphite or semiconductive particles embedded in a polymer matrix. In this study, we employed Co2+ ion-substitution to improve Vis-NIR reflectivity of hydroxyapatite (Ca10(PO4)6(OH)2, HAP) powder. HAP:Co with nominally 5 at.% of Co was prepared with hydrothermal processing of a precipitate. Synthesized powder was characterized by XRD, Raman and ATR-FTIR spectroscopy, FE-SEM and TEM. Thermal stability of HAP:Co powder was examined by simultaneous TG-DTA analyzer. To modify its optical properties and obtain powders with a varietty of color tone, the HAP:Co powder was calcined at 800, 1000, and 1100 °C, in an air atmosphere, for 1 hour. Afterward, the calcined particles were used to prepare composites with poly(vinyl butyral), (PVB); the concentration range was 1 wt.% of the HAP:Co in PVB. The composite coatings, in the form of thin films on glass, were prepared by the solvent-casting technique, using ethanol as a fast evaporating solvent. Firstly, the HAP:Co particles were dispersed in ethanol, then PVB was added (Mowital B30H) and dissolved. To evaporate the solvent before spectrophotometric measurements, the coatings on glass were dried at room temperature for 72 hours. To comprehend optical properties of the coatings, diffuse reflection, transmission, and color coordinates were determined. We found that calcined HAP:Co particles have potential to be used in the formulation of coatings for camouflage protection

Insight in miRNome of severe multiple sclerosis: Pilot study of distinctive relapse-onset MS phenotypes

Molecular background and biomarkers of highly heterogenous and hardly predictable disease progression among relapse-onset MS patients are of high research interest. In the current pilot study, we aimed to employ next-generation sequencing to investigate the expression of whole small non-coding microRNAs (miRNome) in two groups of MS patients with highly distinctive progression phenotype: one with fast progressing, severely disabling course vs. mild course of MS, longitudinally followed 10 years. Peripheral blood mononuclear cells (PBMC) miRNome data was obtained from mild phenotype MS (n=4 patients) and progressive phenotype MS (n=5 patients), using TakaraBio SMARTer smRNA-Seq Kit on iSeq100 (Illumina). Pre-processing of raw sequencing data, quality control and miRNA differential expression analysis was performed using sRNAtoolbox pipeline. Functional interpretation of differentially expressed miRNA target genes was done in DIANA-miRPathv3.0. Tarbase v8.0 served as a resource of miRNA:gene interactions. Achieved read depth was approximately 1 million raw reads/sample, allowing detection of up to 92 mature miRNAs after genome alignment and miRbase v22 annotation. Differential expression analysis identified the significant upregulation of hsa-miR-23c (log2FC=4.29, Padj= 0.03) in progressive phenotype. Top significantly enriched KEGG pathways in hsa-miR-23c targets suggested regulation of molecular pathways involved in autoimmunity (antigen presentation, Epstein-Barr virus infection) and cancer. In conclusion, this pilot study indicates phenotype-related differences in expression of miRNAs, molecules with high regulatory and biomarker properties. Although detected in PBMC, has-miR-23c is highly expressed in the brain and target MS relevant genes such as, HLA (A, B, C), transferrin receptor, Nrf2, recently proposed to play important role in neurodegeneration

Crystal structure, optical properties and photo/electrocatalytic activity of nanostructured Zn1-xFeyO(1-x+1.5y)

Zink oxide-based materials have a great potential to be applied in photo and electro catalysts, opto-electronic (indoor illumination, LED), etc. Attractiveness of ZnO is attributed to wide bandgap energy at room temperature (3.37 eV), high electron mobility and transfer efficiency (115-155 cm2·V-1·s-1), large exciton binding energy (60 meV), intrinsic stability, nontoxicity, environmental compatibility and also, simple and not expensive synthesis procedure. A lot of different approaches can be used to modify the bandgap (i.e. optical absorption) of ZnO materials: metal and nonmetal ion doping, hydrogenation, the incorporation of crystalline defects in the form of V and I, modification of particles morphology and surface topology, etc. In this study, eco-friendly and rapid microwave processing of a precipitate was used to produce Fe-doped ZnO nanoparticles with 5, 10, 15 and 20 at.% of Fe (Zn1-xFeyO(1-x+1.5y)). The influence of different amount of Fe substituted Zn in ZnO on the crystal structure, morphological, textural, and optical properties as well as on functionality of ZnO particles was investigated. The crystal structure and phase purity of the Zn1-xFeyO(1-x+1.5y) particles were determined by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy and X-ray photoelectron spectroscopy (XPS). Effects of the Fe3+ amount on particles morphology and texture properties were observed with field emission scanning electron microscopy (FE–SEM), transmission electron microscopy (TEM) and nitrogen adsorption–desorption isotherm, respectively. Optical properties were studied using UV-Vis diffuse reflectance and photoluminescence spectroscopy. Functionality of ZnO particles was studied due to their photocatalytic and electrochemical activities. Photocatalytic activity was examined via decolorization of methylene blue under direct sunlight irradiation. Electrochemical behavior of the ZnO samples as anode material was evaluated by linear sweep voltammetry in 0.5 M Na2SO4 electrolyte

Crystal structure, optical properties and photo/electrocatalytic activity of nanostructured Zn1-xFeyO(1-x+1.5y)

Zink oxide-based materials have a great potential to be applied in photo and electro catalysts, opto-electronic (indoor illumination, LED), etc. Attractiveness of ZnO is attributed to wide bandgap energy at room temperature (3.37 eV), high electron mobility and transfer efficiency (115-155 cm2·V-1·s-1), large exciton binding energy (60 meV), intrinsic stability, nontoxicity, environmental compatibility and also, simple and not expensive synthesis procedure. A lot of different approaches can be used to modify the bandgap (i.e. optical absorption) of ZnO materials: metal and nonmetal ion doping, hydrogenation, the incorporation of crystalline defects in the form of V and I, modification of particles morphology and surface topology, etc. In this study, eco-friendly and rapid microwave processing of a precipitate was used to produce Fe-doped ZnO nanoparticles with 5, 10, 15 and 20 at.% of Fe (Zn1-xFeyO(1-x+1.5y)). The influence of different amount of Fe substituted Zn in ZnO on the crystal structure, morphological, textural, and optical properties as well as on functionality of ZnO particles was investigated. The crystal structure and phase purity of the Zn1-xFeyO(1-x+1.5y) particles were determined by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy and X-ray photoelectron spectroscopy (XPS). Effects of the Fe3+ amount on particles morphology and texture properties were observed with field emission scanning electron microscopy (FE–SEM), transmission electron microscopy (TEM) and nitrogen adsorption–desorption isotherm, respectively. Optical properties were studied using UV-Vis diffuse reflectance and photoluminescence spectroscopy. Functionality of ZnO particles was studied due to their photocatalytic and electrochemical activities. Photocatalytic activity was examined via decolorization of methylene blue under direct sunlight irradiation. Electrochemical behavior of the ZnO samples as anode material was evaluated by linear sweep voltammetry in 0.5 M Na2SO4 electrolyte.IX International School and Conference on Photonics : PHOTONICA2023 : book of abstracts; August 28 - September 1, 2023; Belgrad

Leptin signalling genetic variants and insulin resistance in multiple sclerosis patients

Last decade provided multiple evidence that link disturbances in metabolic processes and energy metabolism with diseases of central nervous system and neurodegeneration. Initial phases of insulin resistance (IR) are present in natural course of multiple sclerosis (MS) and leptin was recognized as a player in MS pathophysiology and moreover cognitive decline. We aimed to investigate association of genetic variants in leptin (LEP) rs7799039, its receptor LEPR rs1137101 and proliferator-activated receptor gamma co-activator 1-alpha (PGCA1A) rs8192678 with IR parameters (HOMA-IR index, area under the curve for insulin and glucose, Cederholm insulin sensitivity index (ISIced), the insulinogenic index in the first 30 min of oral glucose tolerance test (OGTT) in patients with MS. Seventy eight relapsing-remitting patients in clinical remission, free of corticosteroids for at least three months, were included in the study. None of the 3 variants’ genotypes were associated with HOMA-IR index, area under the curve for insulin and glucose and the insulinogenic index in the first 30 min of OGTT. PGC1A variant was significantly associated with ISIced (Kruskal-Wallis ANOVA, p = 0.04). Leptin gene variant was significantly associated with impaired GT (p=0.029 adjusted for gender and other two variants). None of the variant showed association with IR. In conclusion, we found that genetic variants in leptin signalling pathway affect glucose tolerance and insulin sensitivity in patients with MS. As both, leptin and PGC1A have role in preventing neuronal death and reducing oxidative stress neuronal damage current results favour further investigation toward preserving cognitive status and neuroprotection in MS.17th World Congress on Controversies in Neurology; March 23-25, Dubrovnik, Croatia, 2023

Association study of rs7799039, rs1137101 and rs8192678 gene variants with disease susceptibility/severity and corresponding LEP, LEPR and PGC1A gene expression in multiple sclerosis

Background: Leptin (LEP), leptin receptor (LEPR) and peroxisome proliferator-activated receptor gamma co-activator 1-alpha (PGC1A) are involved in the pathogenesis of multiple sclerosis (MS) by affecting the inflammatory response and reactive oxygen species production. LEP rs7799039 and LEPR rs1137101 genetic variants modify the serum LEP levels and PGC1A rs8192678 alters the PGC1A activity. The study objective was to explore the associations of these variants with susceptibility to MS, disease course/clinical parameters and also with peripheral blood mononuclear cell expression of the target genes and plasma LEP concentrations, in the study subjects. Methods: The study groups included 528 patients with MS and 429 controls. TaqMan® assays were used for genotyping and gene expression quantification. The Chi-square, parametric and nonparametric tests and simple/multiple logistic regression were performed for the statistical analysis of data. Results: A multiple logistic regression model including all three investigated variants, applied to patients (RRMS + SPMS) and controls, showed that PGC1A rs8192678 minor allele had an increased risk for the occurrence of disease, with OR (95%CI) = 1,32 (1,01–1,73), P = 0,04. Between-effect of gender and LEPR variant on the multiple sclerosis severity score (MSSS) was identified (P = 0,005). In male patients (relapsing-remitting and secondary progressive), LEPR minor allele carriers had increased MSSS (GG + AG vs AA, median (minimum–maximum) = 5,38 (0,64–9,88) vs 4,27 (0,78–9,63); P = 0,01, Padj = 0,03). In relapsing-remitting patients LEP rs7799039 affected the LEP gene expression (P = 0,006; Padj = 0,04). Conclusion: The current findings implicate an impact of investigated genetic variants on the pathogenesis of MS. © 2021 Elsevier B.V

The glutathione S-transferase T1 deletion is associated with susceptibility to multiple sclerosis

Background: Multiple sclerosis (MS) occurs as a result of interaction between genetic and environmental factors. Recent data support the view that oxidative damage is one of an early event in MS tissue injury. The safe elimination of reactive oxygen species and toxins via glutathione S-transferase (GST) pathways is required in order to protect cells against reactive oxygen-induced damage. The aim of our study was to analyze the possible association of GSTM1 and GSTT1 gene polymorphisms with the susceptibility and clinical parameters of MS, in 455 consecutive patients and 366 controls. Methods: A multiplex polymerase chain reaction (PCR) was used to detect the deletions in GSTM1 and GSTT1 genes. Results: Patients with MS had significantly higher frequency of GSTT1 null genotype compared to controls (37.36% vs. 21.86%, respectively, p LT 0.0001, adjusted OR 2.13 (1.56-2.90)), as well as double deletions (15.38% vs. 10.38%, respectively, p LT 0.05). The carriers of GSTM1 deletion had significantly earlier onset of MS compared to the wild-type carriers (28.31 +/- 8.45 vs. 30.64 +/- 9.30 years, respectively, p = 0.03). Conclusion: This study suggests the potential pathogenic role of GSTT1 deletion on MS susceptibility. There are no similar data published so far, yet this study should be replicated in other populations. (C) 2013 Elsevier B.V. All rights reserved