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..

Varijante i transkripcija gena koji kodiraju komponente leptinskog signalnog puta, inflamacije i antioksidativne zaštite u patogenezi multiple skleroze

Multipla skleroza (MS) je hronična autoimunska bolest centralnog nervnog sistema (CNS) čiji osnovni patogenetski mehanizmi obuhvataju neuroinflamaciju, demijelinizaciju i neurodegeneraciju. Glavno patofiziološko obeležje MS je hiperaktivacija određenih komponenti imunskog sistema, koja dovodi do nastajanja i akumulacije strukturno-funkcionalnih oštećenja CNS kao posledice hronične inflamacije i oksidativnog stresa. Adipocitokin i hormon leptin potencijalni je molekularni marker MS, budući da učestvuje u patogenezi ove bolesti delovanjem na regulaciju imunskog i odgovora na oksidativni stres. Varijante i promene u transkripciji gena koji kodiraju komponente leptinskog signalnog puta, i sa njima povezane komponente inflamacije i antioksidativne zaštite, asocirane su sa nastankom i/ili kliničkim tokom MS kao i odgovorom na terapiju. Danas u Srbiji ima preko 9000 obolelih od MS, a samo 1 od 10 pacijenata dobija trenutno dostupnu konvencionalnu imunomodulatornu terapiju. Mogućnost modulisanja nivoa cirkulišućeg leptina sugeriše njegov potencijalni značaj u budućem razvoju personalizovanog pristupa u lečenju MS baziranog na poboljšavanju efekata ili supstituciji postojeće konvencionalne terapije.Multiple sclerosis (MS) is a chronic autoimmune disease of the central nervous system (CNS) with underlying pathogenetic mechanisms that include neuroinflammation, demyelination and neurodegeneration. The main pathophysiological feature of MS is the hyperactivation of certain immune system components, which leads to formation and accumulation of structural and functional damage in the CNS as a result of chronic inflammation and oxidative stress. The adipocytokine and hormone leptin is a potential molecular marker of MS, since it participates in the pathogenesis of this disease by acting on the regulation of immune system and response to oxidative stress. Variants and transcriptional changes of genes coding for components of the leptin signaling pathway, and associated components of inflammation and antioxidant protection, are associated with the onset and/or clinical course of MS as well as response to therapy. Today, there are over 9,000 MS patients in Serbia, and only 1 out of 10 patients receives currently available conventional immunomodulatory therapy. The possibility of modulating the level of circulating leptin suggests its potential importance in the future development of a personalized approach in treatment of MS based on improving the effects or substituting the existing conventional therapy

Community Pharmacists' Attitudes and Professional Practice in Relation to the Patient Safety Incidents

Background: Medicines dispensing is an error-prone activity, therefore potentially jeopardizing patient safety. This study aimed to assess the community pharmacists' attitudes towards the causes of dispensing errors and preventive measures, as well as their practice in incidents reporting. Materials and Methods: A cross-sectional survey was performed by distributing an adopted and validated questionnaire to a nationwide sample of community pharmacists in Serbia. The questionnaire included sections related to the participants' socio-demographic characteristics, their attitudes towards factors causing dispensing errors and corrective actions, as well as their practice in reporting. Statistical analyses were conducted using SPSS Statistics software ver. 21.0. The associations between categorical variables were analyzed using Chi-square test. Results: The study included 1,004 participants, mainly female (94.9%), with the mean age 40.9 +/- 9.9 years and mean work experience 14.3 +/- 10.0 years. More than a third of the participants (35.4%) indicated an increasing risk of dispensing errors. The main causes included illegible prescriber's handwriting (44.3%) and interruptions during dispensing (39.2%), while the major corrective actions were providing pharmacists with education in clinical pharmacy (71%) and reducing the interruptions during dispensing (63.9%). The majority of respondents (85.2%) stated that they routinely reported dispensing incidents. However, even 16.5% of them admitted to having fear sometimes or always. Additionally, only 58.1% of participants would use voluntary dispensing error reporting system. Conclusion: Serbian community pharmacists are aware of the existing risk in medicines dispensing and the corrective actions identified should be put into practice so as to manage them prospectively. Although the results indicate good practice in incidents reporting, conducting tailored educations and building of safety culture is necessary to improve patient safety

Polyphenol-Rich Aronia melanocarpa Juice Consumption Affects LINE-1 DNA Methylation in Peripheral Blood Leukocytes in Dyslipidemic Women

Cardiovascular disease (CVD) is associated with alterations in DNA methylation and polyunsaturated fatty acid (PUFA) profile, both modulated by dietary polyphenols. The present parallel, placebo-controlled study (part of the original clinical study registered as NCT02800967 at www.clinicaltrials.gov) aimed to determine the impact of 4-week daily consumption of polyphenol-rich Aronia melanocarpa juice (AMJ) treatment on Long Interspersed Nucleotide Element-1 (LINE-1) methylation in peripheral blood leukocytes and on plasma PUFAs, in subjects (n = 54, age range of 40.2 ± 6.7 years) at moderate CVD risk, including an increased body mass index, central obesity, high normal blood pressure, and/or dyslipidemia. The goal was also to examine whether factors known to affect DNA methylation (folate intake levels, MTHFR C677T gene variant, anthropometric and metabolic parameters) modulated the LINE-1 methylation levels upon the consumption of polyphenol-rich aronia juice. Experimental analysis of LINE-1 methylation was done by MethyLight method. MTHFR C677T genotypes were determined by the polymerase chain reaction–restriction fragment length polymorphism method, and folate intake was assessed by processing the data from the food frequency questionnaire. PUFAs were measured by gas–liquid chromatography, and serum lipid profile was determined by using Roche Diagnostics kits. The statistical analyses were performed using Statistica software package. In the comparison after vs. before the treatment period, in dyslipidemic women (n = 22), we observed significant decreases in LINE-1 methylation levels (97.54 ± 1.50 vs. 98.39 ± 0.86%, respectively; P = 0.01) and arachidonic acid/eicosapentaenoic acid ratio [29.17 ± 15.21 vs. 38.42 (25.96–89.58), respectively; P = 0.02]. The change (after vs. before treatment) in LINE-1 methylation directly correlated with the presence of MTHFR 677T allele, average daily folate intake, and the change in serum low-density lipoprotein cholesterol but inversely correlated with the change in serum triacylglycerols (R = 0.72, R2 = 0.52, adjusted R2 = 0.36, P = 0.03). The current results imply potential cardioprotective effects of habitual polyphenol-rich aronia juice consumption achieved through the modifications of DNA methylation pattern and PUFAs in subjects at CVD risk, which should be further confirmed. Hence, the precision nutrition-driven modulations of both DNA methylation and PUFA profile may become targets for new approaches in the prevention of CVD

Is There a FADS2-Modulated Link between Long-Chain Polyunsaturated Fatty Acids in Plasma Phospholipids and Polyphenol Intake in Adult Subjects Who Are Overweight?

Dietary polyphenols promote cardiometabolic health and are linked with long-chain polyunsaturated fatty acids in plasma phospholipids (LC-PUFA). The FADS2 polymorphisms are associated with LC-PUFA metabolism and overweight/obesity. This 4-week study examined the link between polyphenol intake, FADS2 variants (rs174593, rs174616, rs174576) and obesity in 62 overweight adults (BMI gt = 25), allocated to consume 100 mL daily of either: Aronia juice, a rich source of polyphenols, with 1177.11 mg polyphenols (expressed as gallic acid equivalents)/100 mL (AJ, n = 22), Aronia juice with 294.28 mg polyphenols/100 mL (MJ, n = 20), or nutritionally matched polyphenol-lacking placebo as a control (PLB, n = 20). We analyzed LC-PUFA (% of total pool) by gas chromatography and FADS2 variants by real-time PCR. Four-week changes in LC-PUFA, BMI, and body weight were included in statistical models, controlling for gender and PUFA intake. Only upon AJ and MJ, the presence of FADS2 variant alleles affected changes in linoleic, arachidonic, and eicosapentaenoic acid (EPA). Upon MJ treatment, changes in EPA were inversely linked with changes in BMI (beta= -0.73, p = 0.029) and weight gain (beta= -2.17, p = 0.024). Only in subjects drinking AJ, the link between changes in EPA and anthropometric indices was modified by the rs174576 variant allele. Our results indicate the interaction between FADS2, fatty acid metabolism, and polyphenol intake in overweight subjects

Dental health care for children with autism spectrum disorders

Introduction: Autism spectrum disorders (ASD) are pervasive disorders beginning in early childhood. The prevalence of these disorders has been on the rise over the last few decades. A disorder of this kind does not have any direct impact on the oral health, but the behavior of the affected children can markedly deteriorate it. Due to a poor compliance to oral hygiene and maintenance of oral health, dental film accumulates in the oral cavity, leading to both caries and its complications, and gingival-periodontal diseases. A high percentage of children with ASD are not sufficiently compliant during dental interventions, being hyperactive, tense, and often highly agitated. Any dental intervention in such children is therefore very complicated or it cannot be performed at all, consequentially broadening the spectrum of indications for dental extractions. In order to preserve the oral health in children with ASD, an individualized approach to each patient is thus necessary. Conclusion: The journey to a healthy oral cavity and teeth in children with ASD is full of twists and turns, but the desired goals can be realized, although requiring considerable patience and perseverance from both the parents and pediatric dentists

The biocorrosion activity of ZnO-based materials as biosensors

Due to their biocompatibility, chemical stability, high isoelectric point, electrochemical activity, high electron mobility and ease of synthesis by diverse methods, ZnO-based materials have attracted much interest as materials for biosensors. Its unique properties allow it to be used for single-molecule detection and determining various biomolecules, so it can be potentially utilized as biosensor for medical diagnosis. The materials being used as biosensors require special characteristics including high corrosion resistance. The aim of this research was to investigate biocorrosion properties of ZnO materials in Ringer’s physiological solution as a function of immersion time. ZnO powders were prepared by microwave (MW) processing of a precipitate in the presence of a different amount (5, 10 and 20 wt.%) of two different surfactants, CA and CTAB. The particles crystallinity and phase purity were investigated by X-ray powder diffraction (XRD) and Raman spectroscopy. Fourier-transform infrared (FTIR) spectroscopy was used to analyze surface chemistry. The particles morphology and textural properties were observed with field emission scanning electron microscopy (FE-SEM) and BET. The biocorrosion activity of the materials was measured by potentiodynamic polarization technique. Prepared samples were immersed in Ringer solution for different immersion times ranging from 30 min to 7 days. We found that all examined ZnO samples hаve low biocorrosion activity. Slight differences in biocorrosion activity between the samples are determined by particles morphology, textural properties and surface chemistry influenced by used surfactants

Enhanced photo(electro)catalytic properties of ZnO particles synthesized by CTAB-assisted microwave processing

ZnO/CTAB powder was prepared by microwave processing of a precipitate with the aid of cetyltrimethylammonium bromide (CTAB). The effects of CTAB on the crystal structure, morphology, optical and photo(electro)catalytic properties of ZnO particles were studied. The results showed that CTAB did not influenced crystal structure or phase purity of ZnO. However, even low concentration of CTAB vary particles morphology; cone-like particles were prepared by processing without CTAB, while a mixture of spheroidal and plate-like ZnO particles were produced when 0.001 M CTAB was used. It was found that synthesized ZnO powders have 0.10 eV lower band gap energy then bulk ZnO (3.37 eV). A high photocatalytic activity for decolorization of methylene blue water solution was established after 2 h of sunlight irradiation; efficiency was 100 and 67% for ZnO/CTAB and ZnO, respectively. Electrochemical test showed faster oxygen evolution kinetics when ZnO/CTAB was used as anode material. Enhanced photo(electro)catalytic activities of ZnO/CTAB particles are attributed to better absorption of visible light due to both, larger dimensions and surface sensitization by CTAB

Approaches to improve photo(electro)catalytic properties of ZnO-based materials

Due to their tunable multifunctional properties zinc oxide (ZnO) based materials have attracted extensive scientific and technological attention. Since they combine different properties such as electrochemical activities, chemical and photochemical stability, nontoxicity, biocompatibility, etc. ZnO-based materials have been used in electronics, optoelectronics, biosensing, bioimaging, drug and gene delivery, implants, antimicrobial and anticancer agents. Successful application of ZnO as photoelectrocatalysts arises from its wide band gap (3.37 eV) which can be easily adjusted by different approaches such as: metal and non-metal ion doping, hydrogenation, introducing of crystalline defects, modifying particle morphology and surface chemistry. During the years, to synthesize zinc oxide (ZnO) nanoparticles with improved visible light absorption we have used a fast and environmentally-friendly microwave processing of a precipitate which enable formation of crystalline defects. To further enhance photo(electro)catalytic properties we have employed approaches such as: (1) the incorporation of iron ions into the crystal structure (Zn1-xFexO), (2) sensitization of the particles’ surface with cetyltrimethylammonium bromide, Pluronic F127 and polyethylene oxide, and (3) composites with ruthenium oxide (ZnO/RuO2) and graphene oxide (ZnO/GO and ZnO/rGO). To correlate structural and functional properties, prepared materials were characterized using XRD, FTIR, Raman, UV-Vis DRS, and PL spectroscopy, also FESEM; photocatalytic activity of the samples were tested toward decolorization of methylene blue, while their photoelectrochemical activity for water splitting were tested through linear sweep voltammetry in different electrolytes