Macrophages – The Key Actors in Adipose Tissue Remodeling and Dysfunction

Adipose tissue (AT) is a very important endocrine and paracrine organ that regulates other tissues and organs. Dysfunction of AT leads to a wide range of disorders like obesity, insulin resistance, diabetes mellitus, cardiac disorders, tumors and others. Adipose tissue macrophages (ATMs) are the key actors in AT remodeling and dysfunction. Their role in AT dysfunction is nowadays increasingly investigated, but still their interplay and molecular mechanisms of actions have not been fully elucidated. In this chapter, we summarized the current knowledge about the role of macrophages in AT remodeling, dysfunction and related disorders and indicate the potential directions for future research

Production and characterization of inulinase enzymes from Aspergillus spp. for obtaining of fructooligosaccharides

Ova disertacija se bavi ispitivanjem mogućnosti primene izolata gljive rodaAspergillus spp. za produkciju enzima inulinaznog kompleksa u cilju dobijanjafruktooligosaharida. Razvijen je brz i pouzdan difuzioni test za detekciju proizvođačainulinaznog kompleksa korišćenog u ispitivanju 39 izolata vrste roda Aspergillus, prethodnoidentifikovanih do nivoa vrste, umnožavanjem CaM gena.Izolati su ispitani na mogućnost produkcije mikotoksina na genetskom nivouumnožavanjem biosintetskih klastera gena za fumonizin i ohratoksin, a analitičkim metodamaispitano je prisustvo mikotoksina u enzimskom preparatu. Ukrštanjem rezultata difuzionogenzimskog testa i mogućnosti produkcije mikotoksina odabran je soj identifikovan kaoAspergillus welwitschiae FAW1, koji ne produkuje mikotoksine i potencijalno je dobarproizvođač enzima za dobijanje FOS-ova, što ga čini pogodnim za upotrebu u proizvodnjihrane.Inulinazni enzimski kompleks je produkovan tokom fermentacije gljive na čvrstojpodlozi upotrebom različitih inducibilnih supstrata (tritikale, jerusalimska artičoka i pšeničnemekinje). Svi enzimi su okarakterisani i dobijene su različite aktivnosti u zavisnosti odsupstrata: egzoinulinazna – InuE (2,4 U/mL) i endoinulinazna – InuA (34 U/mL) dobijene suna tritikaleu, dok je najveća β-fruktofuranozidazna – FFase (6,3 U/mL) i fruktoziltransferazna– FTase aktivnost dobijena na podlozi sa jerusalimskom artičokom, pšeničnim mekinjama ipeptonom. Koprodukcijom inulinaznih enzimskih kompleksa pokazano je da se ista gljivamože koristiti za obe metode dobijanja FOS-ova (FOSs i FOSh) u zavisnosti od podloge nakojoj se uzgaja.Razvijena je nova zimogramska metoda za simultanu detekciju enzima InuA, InuE,FFase nakon jednog elektroforetskog razdvajanja enzima.Uspešno su prečišćeni glavni enzimi odgovorni za produkciju FOS-ova – InuA zadobijanje FOSh i FTase za dobijanje FOSs. Potvrđeno je prisustvo gena suc1 u genomu A.welwitschiae FAW1 koji se smatra odgovornim za ekspresiju enzima FTase i FFase. DobijeniFOSs i FOSh poseduju značajan antioksidativni potencijal što ih čini dobrim kandidatima zadodatak funkcionalnoj hrani.This dissertation examines the possibility of using isolates of Aspergillus spp. for theproduction of the inulinase complex enzyme for obtaining of fructooligosaccharides. A rapidand reliable diffusion test was developed for the detection of the inulinase complex enzymeproducers used in the examination of 39 Aspergillus spp. isolates from the black aspergilligroup, previously identified to the species level, by amplification of the CaM gene.The isolates were examined for the possibility of mycotoxin production on the geneticlevel by amplifying the biosynthetic gene clusters for fumonisin and ochratoxin. Thepresence of mycotoxins in the enzyme preparation was examined using analytical methods.By crossing the results of the diffusion enzyme test and the possibility of mycotoxinproduction, a strain identified as Aspergillus welwitschiae FAW1 was selected as apotentially good producer of enzymes for obtaining FOS, which does not producemycotoxins what makes it safe for use in food production.The inulinase enzyme complex was produced during the fermentation of the fungi ona different solid inducible substrates (triticale, Jerusalem artichoke and wheat bran). Allenzymes were characterized and different activities were obtained depending on thesubstrate: exoinulinase - InuE (2.4 U/mL) and endoinulinase - InuA (34 U/mL) were obtainedon triticale, while the highest β-fructofuranosidase - FFase (6.3 U/mL) andfructosyltransferase - FTase activity were obtained on a medium with Jerusalem artichoke,wheat bran and peptone. The co-production of inulinase enzyme complexes showed that thesame fungi can be used for both methods of obtaining FOS (FOSs and FOSh), depending onthe substrate on which it was grown.A new zymographic method for the simultaneous detection of enzymes InuA, InuEand FFase after one electrophoretic separation of the enzymes was developed.The main enzymes responsible for the production of FOS were successfully purified -InuA for obtaining FOSh and FTase for obtaining FOSs. The presence of the suc1 gene in theA. welwitschiae FAW1 genome, which is considered responsible for the expression of FTaseand FFase enzymes, was confirmed. The obtained FOSs and FOSh have significantantioxidant potential, which makes them good candidates for use in functional food

Production and characterization of inulinase enzymes from Aspergillus spp. for obtaining of fructooligosaccharides

Ova disertacija se bavi ispitivanjem mogućnosti primene izolata gljive roda Aspergillus spp. za produkciju enzima inulinaznog kompleksa u cilju dobijanja fruktooligosaharida. Razvijen je brz i pouzdan difuzioni test za detekciju proizvođača inulinaznog kompleksa korišćenog u ispitivanju 39 izolata vrste roda Aspergillus, prethodno identifikovanih do nivoa vrste, umnožavanjem CaM gena. Izolati su ispitani na mogućnost produkcije mikotoksina na genetskom nivou umnožavanjem biosintetskih klastera gena za fumonizin i ohratoksin, a analitičkim metodama ispitano je prisustvo mikotoksina u enzimskom preparatu. Ukrštanjem rezultata difuzionog enzimskog testa i mogućnosti produkcije mikotoksina odabran je soj identifikovan kao Aspergillus welwitschiae FAW1, koji ne produkuje mikotoksine i potencijalno je dobar proizvođač enzima za dobijanje FOS-ova, što ga čini pogodnim za upotrebu u proizvodnji hrane. Inulinazni enzimski kompleks je produkovan tokom fermentacije gljive na čvrstoj podlozi upotrebom različitih inducibilnih supstrata (tritikale, jerusalimska artičoka i pšenične mekinje). Svi enzimi su okarakterisani i dobijene su različite aktivnosti u zavisnosti od supstrata: egzoinulinazna – InuE (2,4 U/mL) i endoinulinazna – InuA (34 U/mL) dobijene su na tritikaleu, dok je najveća β-fruktofuranozidazna – FFase (6,3 U/mL) i fruktoziltransferazna – FTase aktivnost dobijena na podlozi sa jerusalimskom artičokom, pšeničnim mekinjama i peptonom. Koprodukcijom inulinaznih enzimskih kompleksa pokazano je da se ista gljiva može koristiti za obe metode dobijanja FOS-ova (FOSs i FOSh) u zavisnosti od podloge na kojoj se uzgaja. Razvijena je nova zimogramska metoda za simultanu detekciju enzima InuA, InuE, FFase nakon jednog elektroforetskog razdvajanja enzima. Uspešno su prečišćeni glavni enzimi odgovorni za produkciju FOS-ova – InuA za dobijanje FOSh i FTase za dobijanje FOSs. Potvrđeno je prisustvo gena suc1 u genomu A. welwitschiae FAW1 koji se smatra odgovornim za ekspresiju enzima FTase i FFase. Dobijeni FOSs i FOSh poseduju značajan antioksidativni potencijal što ih čini dobrim kandidatima za dodatak funkcionalnoj hrani.This dissertation examines the possibility of using isolates of Aspergillus spp. for the production of the inulinase complex enzyme for obtaining of fructooligosaccharides. A rapid and reliable diffusion test was developed for the detection of the inulinase complex enzyme producers used in the examination of 39 Aspergillus spp. isolates from the black aspergilli group, previously identified to the species level, by amplification of the CaM gene. The isolates were examined for the possibility of mycotoxin production on the genetic level by amplifying the biosynthetic gene clusters for fumonisin and ochratoxin. The presence of mycotoxins in the enzyme preparation was examined using analytical methods. By crossing the results of the diffusion enzyme test and the possibility of mycotoxin production, a strain identified as Aspergillus welwitschiae FAW1 was selected as a potentially good producer of enzymes for obtaining FOS, which does not produce mycotoxins what makes it safe for use in food production. The inulinase enzyme complex was produced during the fermentation of the fungi on a different solid inducible substrates (triticale, Jerusalem artichoke and wheat bran). All enzymes were characterized and different activities were obtained depending on the substrate: exoinulinase - InuE (2.4 U/mL) and endoinulinase - InuA (34 U/mL) were obtained on triticale, while the highest β-fructofuranosidase - FFase (6.3 U/mL) and fructosyltransferase - FTase activity were obtained on a medium with Jerusalem artichoke, wheat bran and peptone. The co-production of inulinase enzyme complexes showed that the same fungi can be used for both methods of obtaining FOS (FOSs and FOSh), depending on the substrate on which it was grown. A new zymographic method for the simultaneous detection of enzymes InuA, InuE and FFase after one electrophoretic separation of the enzymes was developed. The main enzymes responsible for the production of FOS were successfully purified - InuA for obtaining FOSh and FTase for obtaining FOSs. The presence of the suc1 gene in the A. welwitschiae FAW1 genome, which is considered responsible for the expression of FTase and FFase enzymes, was confirmed. The obtained FOSs and FOSh have significant antioxidant potential, which makes them good candidates for use in functional food

INVESTMENT PROJECT SELECTION BY APPLYING COPRAS METHOD AND IMPRECISE DATA

Investment projects can have a significant impact on the functioning and development of acompany. Therefore, the selection of one or more investment projects from the set of possible is animportant and difficult task for decision makers. This paper considers the investment projectsselection based on financial analysis criteria and use of imprecise data. In the proposed model, thealternative projects performances are expressed using crisp and interval values, and then the bestproject from the available is selected by using COPRAS and COPRAS-G methods. A numericalexample is given to demonstrate the applicability and effectiveness of the proposed approach

Production and characterization of inulinase enzymes from Aspergillus spp. for obtaining of fructooligosaccharides

Ova disertacija se bavi ispitivanjem mogućnosti primene izolata gljive rodaAspergillus spp. za produkciju enzima inulinaznog kompleksa u cilju dobijanjafruktooligosaharida. Razvijen je brz i pouzdan difuzioni test za detekciju proizvođačainulinaznog kompleksa korišćenog u ispitivanju 39 izolata vrste roda Aspergillus, prethodnoidentifikovanih do nivoa vrste, umnožavanjem CaM gena.Izolati su ispitani na mogućnost produkcije mikotoksina na genetskom nivouumnožavanjem biosintetskih klastera gena za fumonizin i ohratoksin, a analitičkim metodamaispitano je prisustvo mikotoksina u enzimskom preparatu. Ukrštanjem rezultata difuzionogenzimskog testa i mogućnosti produkcije mikotoksina odabran je soj identifikovan kaoAspergillus welwitschiae FAW1, koji ne produkuje mikotoksine i potencijalno je dobarproizvođač enzima za dobijanje FOS-ova, što ga čini pogodnim za upotrebu u proizvodnjihrane.Inulinazni enzimski kompleks je produkovan tokom fermentacije gljive na čvrstojpodlozi upotrebom različitih inducibilnih supstrata (tritikale, jerusalimska artičoka i pšeničnemekinje). Svi enzimi su okarakterisani i dobijene su različite aktivnosti u zavisnosti odsupstrata: egzoinulinazna – InuE (2,4 U/mL) i endoinulinazna – InuA (34 U/mL) dobijene suna tritikaleu, dok je najveća β-fruktofuranozidazna – FFase (6,3 U/mL) i fruktoziltransferazna– FTase aktivnost dobijena na podlozi sa jerusalimskom artičokom, pšeničnim mekinjama ipeptonom. Koprodukcijom inulinaznih enzimskih kompleksa pokazano je da se ista gljivamože koristiti za obe metode dobijanja FOS-ova (FOSs i FOSh) u zavisnosti od podloge nakojoj se uzgaja.Razvijena je nova zimogramska metoda za simultanu detekciju enzima InuA, InuE,FFase nakon jednog elektroforetskog razdvajanja enzima.Uspešno su prečišćeni glavni enzimi odgovorni za produkciju FOS-ova – InuA zadobijanje FOSh i FTase za dobijanje FOSs. Potvrđeno je prisustvo gena suc1 u genomu A.welwitschiae FAW1 koji se smatra odgovornim za ekspresiju enzima FTase i FFase. DobijeniFOSs i FOSh poseduju značajan antioksidativni potencijal što ih čini dobrim kandidatima zadodatak funkcionalnoj hrani.This dissertation examines the possibility of using isolates of Aspergillus spp. for theproduction of the inulinase complex enzyme for obtaining of fructooligosaccharides. A rapidand reliable diffusion test was developed for the detection of the inulinase complex enzymeproducers used in the examination of 39 Aspergillus spp. isolates from the black aspergilligroup, previously identified to the species level, by amplification of the CaM gene.The isolates were examined for the possibility of mycotoxin production on the geneticlevel by amplifying the biosynthetic gene clusters for fumonisin and ochratoxin. Thepresence of mycotoxins in the enzyme preparation was examined using analytical methods.By crossing the results of the diffusion enzyme test and the possibility of mycotoxinproduction, a strain identified as Aspergillus welwitschiae FAW1 was selected as apotentially good producer of enzymes for obtaining FOS, which does not producemycotoxins what makes it safe for use in food production.The inulinase enzyme complex was produced during the fermentation of the fungi ona different solid inducible substrates (triticale, Jerusalem artichoke and wheat bran). Allenzymes were characterized and different activities were obtained depending on thesubstrate: exoinulinase - InuE (2.4 U/mL) and endoinulinase - InuA (34 U/mL) were obtainedon triticale, while the highest β-fructofuranosidase - FFase (6.3 U/mL) andfructosyltransferase - FTase activity were obtained on a medium with Jerusalem artichoke,wheat bran and peptone. The co-production of inulinase enzyme complexes showed that thesame fungi can be used for both methods of obtaining FOS (FOSs and FOSh), depending onthe substrate on which it was grown.A new zymographic method for the simultaneous detection of enzymes InuA, InuEand FFase after one electrophoretic separation of the enzymes was developed.The main enzymes responsible for the production of FOS were successfully purified -InuA for obtaining FOSh and FTase for obtaining FOSs. The presence of the suc1 gene in theA. welwitschiae FAW1 genome, which is considered responsible for the expression of FTaseand FFase enzymes, was confirmed. The obtained FOSs and FOSh have significantantioxidant potential, which makes them good candidates for use in functional food

Povezanost nivoa adiponektina u serumu pacijenata u zavisnosti od metaboličkog sindroma i koronarne bolesti

Dosadašnje studije potvrđuju da adiponektin, protein koji luče adipociti, ostvaruje kardiovaskuloprotektivnu, antiaterogenu, anti- inflamatornu, antiadhezivnu, vazodilatatornu, antikataboličku i antidijabetičnu ulogu. Hipoadiponektinemija doprinosi patofiziologiji metaboličkog sindroma i koronarne bolesti, čak može biti nezavistan kardiovaskularni faktor rizika. Nivo adiponektina u serumu može biti rani marker ovih entiteta, metaboličkog sindroma i koronarne bolesti. Metod: Kliničko-laboratorijska studija preseka obuhvatila je 100 ispitanika oba pola sa metaboličkim sindromom i koronarnom bolešću. Korišćene su sledeće metode: klinički pregled, antropometrijska merenja, određivanje ukupne količne masnog tkiva osteodenzitometrijom, labora- torijska ispitivanja [koncentracija adiponektina u serumu ELISA metodom, insulina RIA metodom, glikemija OGTT-om, lipidni parametri (HOL, HDL, LDL, trigliceridi) i C-reaktivni protein], elektrokardiogramsko, ehokardiografsko merenje, analiza koronarografskih nalaza i izračunavanje aterogenih i HOMA-indeksa. Rezultati: Nivo adiponektina je u negativnoj korelaciji sa parametrima metaboličkog sindroma (gojaznost, hipertenzija, hiperglikemija i insulinska rezistencija, dislipidemija) i koronarne bolesti (broj suženja koronarnih arterija i progresija bolesti) i u pozitivnoj korelaciji sa ejekcijonom frakcijom, dijastolnom funkcijom i HDL-holesterolom. Adiponektin može biti značajan prediktor metaboličkog sindroma i koronarne bolesti i rani reprezentativni marker miokardne ishemije i vaskularne inflamacije, a hipoadiponektinemija nezavistan kardiovaskularni faktor rizika. Zaključak: Studija, prva te vrste u Srbiji, a jedna od retkih u svetu, daće značajan doprinos izučavanju adiponektina kao ranog reprezentativnog, dijagnostičkog i prediktivnog markera kod bolesnika sa metaboličkim sindromom i koronarnom bolešću.Up to date studies are confirming that adiponectin, a cytokine produced by adipocytes, exerts cardiovascular-protective role including antiatherogenic, anti-inflammatory, anti-adherent, vasodilator, anticatabolic and antidiabetic effects. Hypoadiponectinaemia contributes to the pathophysiology of the metabolic syndrome and coronary artery disease. The level of adiponectin in serum may be seen as an independent cardiovascular risk factor. In addition, the level of circulating adiponectin may be an early marker of the conditions such as metabolic syndrome and coronary artery disease. Methods: This research represents the clinical-laboratory cross-sectional study, based on the cases of 100 patients of both sexes with metabolic syndrome and/or coronary artery disease. Following methods were used: a clinical examination, anthropometric measurements, determine the total amounts of the fatty tissue osteodensitometry; laboratory testing included concentration of adiponectin in serum utilising super-sensitive sandwich ELISA-method, insulin level measured by RIA-method, glycemia OGTT-test, lipid parameters (HOL, HDL, LDL, triglycerides) and C-reactive protein, gained by standard routine biochemical protocols. All of the examinees were subjected to electrocardiogram and ehocardiogram measurement, the analysis of graphs-coronary graphically findings and calculation of atherogenic and HOMA-index. Results: The level of adiponectin showed statisticaly significant negative correllation with the key parameters of metabolic syndrome (obesity, hypertension, hyperglycemia, dyslipidemia and insulin resistance), as well as coronary artery disease. More accurately, the level of adiponectin is decreasing exact number of coronary arteries disease and simptomatology progression. Plasma levels of adiponectin positively correlated with ejection fraction, diastolic function and HDL-cholesterol. Adiponectin may be a significant predictor of metabolic syndrome and coronary heart disease, and an early representative marker of myocardial ischemia and vascular inflammation, as well as hypoadiponectinaemia independent cardiovascular risk factor. Conclusion: This study, the first of its kind in Serbia and one of the few in the world, will significantly contribute to acknowledgement of the role of adiponectin as an early representative, diagnostic and predictive markers in patients with metabolic syndrome and coronary artery disease

Morphological and molecular characterization of human adipose tissue and lipomas of different anatomical localization and examination of isolated mesenchymal cells in vitro

Lipomi su benigni tumori masnog tkiva sa još uvek nedovoljno razjašnjenom etiologijom i patogenezom. Lipomi su uglavnom lokalizovani potkožno ali se mogu naći i intramuskularno, u telesnim šupljinama i organima, mogu biti solitarni ili multipli, mogu varirati u veličini i histološkoj građi, ali podaci o njihovom molekularnom profilu i mehanizmima nastanka su vrlo oskudni. Mezenhimske matične ćelije iz lipoma (eng. lipomaderived stem cells - LDSCs) su predložene za primenu u tkivnom inženjerstvu i regenerativnoj medicini od strane autora nekoliko studija zato što je pokazano da imaju karakteristike slične mezenhimskim matičnim ćelijama iz masnog tkiva (eng. adipose-derived stem cells - ADSCs). Osim podataka o potencijalu izolovanih LDSCs da proliferišu i da se diferentuju u adipocite, osteoblaste i hondrocite, nema podataka o ostalim karakteristikama ovih ćelija a ni detaljnog poređenja sa ADSCs na ćelijskom i molekularnom nivou. Cilj jednog dela ovog istraživanja je bio ispitivanje molekularnog profila izolovanih LDSCs i poređenje sa ADSCs u kulturi in vitro, ispitivanje kapaciteta LDSCs da se diferentuju u adipocite i osteoblaste, na ćelijskom, proteinskom i genskom nivou, ispitivanje uticaja kondicioniranih medijuma LDSCs na odgovor makrofaga, kao i ispitivanje imunomodulacijske aktivnosti i efekta na zarastanje rana u sistemu indirektne ko-kulture in vitro, u poređenju sa ADSCs. Cilj drugog dela ovog istraživanja je bio morfološka i molekularna karakterizacija tkiva lipoma i potkožnog belog masnog tkiva (eng. subcutaneous white adipose tissue – scWAT) različitih anatomskih lokalizacija. Rezultati su pokazali da LDSCs i ADSCs poseduju karakteristike mezenhimskih matičnih ćelija i da imaju sličan fenotip ali različitu molekularnu osnovu za diferencijaciju. Obrazac ekspresije gena markera adipogeneze i prisustvo manjeg broja zrelih adipocita u kulturi LDSCs u odnosu na ADSCs nakon 21 dan adipogene diferencijacije ukazuju na slabiji kapacitet LDSCs za diferencijaciju u adipocite u poređenju sa ADSCs. Analiza osteogenih markera nakon 16 dana osteogene diferencijacije je pokazala da i LDSCs i ADSCs imaju fenotip karakterističan za osteoblaste, ali su bile u različitim fazama osteogeneze. Kondicionirani medijumi LDSCs (LDSC-CM) i ADSCs (ADSC-CM) dovode do funkcijske aktivacije nestimulisanih makrofaga i ispoljavaju anti-inflamacijsku aktivnost, sa izraženijim efektom LDSC-CM. LDSC-CM i ADSC-CM deluju povoljno na zarastanje rana u sistemu indirektne ko-kulture in vitro, bez značajnih razlika, i u nivou su pozitivne kontrole...Lipomas are benign adipose tissue tumors with still insufficiently clarified etiology and pathogenesis. Lipomas are mainly localized subcutaneously but can also be found intramuscularly, in the body cavities and organs, may be solitary or multiple, may vary in size and histological structure, but data on their molecular profile and mechanisms of formation are still scarce. Lipoma-derived stem cells (LDSCs) have been proposed for use in tissue engineering and regenerative medicine by the authors of several studies since it has been shown that these cells have characteristics similar to adipose-derived stem cells (ADSCs). In addition to data on the potential of isolated LDSCs to proliferate and differentiate into adipocytes, osteoblasts and chondrocytes, there is no data on other characteristics of these cells, nor a detailed comparison with ADSCs at the cellular and molecular levels. The aim of one part of this study was to examine the molecular profile of isolated LDSCs and comparison with ADSCs in culture in vitro, to examine the capacity of LDSCs to differentiate into adipocytes and osteoblasts on cellular, protein and gene level, to examine the influence of conditioned media of LDSCs on macrophages’ response, as well as to examine immunomodulatory activity and effect of wound healing in indirect coculture system in vitro, in comparison with ADSCs. The aim of the second part of this study was the morphological and molecular characterization of lipoma tissue and subcutaneous white adipose tissue (scWAT) of various anatomical localizations. The results showed that LDSCs and ADSCs possess the characteristics of mesenchymal stem cells and have similar phenotype but different molecular basis for differentiation. The pattern of expression of genes that are markers of adipogenesis and the presence of less number of mature adipocytes in LDSCs compared to ADSCs culture after 21 day of adipogenic differentiation, indicate lower capacity of LDSCs for differentiation into adipocytes compared to ADSCs. Analysis of osteogenic markers after 16 days of osteogenic differentiation showed that both LDSCs and ADSCs have a characteristic osteoblast-like phenotype, but they were at different stages of osteogenesis..

Prescribing antibiotics to preschool children in primary health care in Croatia

The use of antibiotics depends on cultural and socioeconomic factors, physician's characteristics as well as on microbiological considerations. Aim of our study was to asses antibiotic prescription among preschool children in primary health care in Croatia in relation to socioeconomic factors, symptoms and diagnoses, and type of health care provider. Retrospective longitudinal survey was conducted in 7 teaching primary health care offices in the Croatian capital of Zagreb during 2004, among 1700 preschool children. Antibiotics were prescribed to 611 (46%) children. Significantly more antibiotics were prescribed to boys (66.7%, P = 0.024) and to children whose parents had lower educational level. Most frequently antibiotics were prescribed for the symptoms such as fever (32%), cough (32.5%), nasal discharge (12%), and for the diagnoses such as respiratory diseases (J00-J99) (40%), infectious and parasitic diseases (A00-A99) (31%), and diseases of the middle ear and mastoid (H60-H95) (15%). Logistic regression analyses also predicted correlation of antibiotic prescriptions with socioeconomic factors, symptoms and diagnoses and health care of pediatrician. Prescription of antibiotics for preschool children in primary health care in Croatia related to socioeconomic factors, type of health care provider, certain symptoms and diagnosis groups which should be taken into account when assessing and planning primary health care for preschool children

COMPARATIVE REVIEW OF GINGIVAL RETRACTION AGENTS

In order to obtain an adequate impression of demarcation line area of a prepared tooth located in or below the gingival edge it is necessary to perform the dilatation and drying of the gingival sulcus using retraction cord impregnated with adequate retraction agents. The aim of the study was to carry out comparative analysis of advantages and disadvantages of commercially available gingival retraction agents. Commercial retraction agents include astringents (metal salts) and vasoconstrictors on the basis of epinephrine. Further research should be aimed at examining the possibility of using sympathomimetic vasoconstrictors (tetrahydrosolin and oxymetasolin) for gingival retraction

Osteogenic differentiation of dental pulp stem cells influenced by synthesized calcium phosphate-based nanomaterial in vitro

Dental pulp stem cells (DPSCs) are mesenchymal stem cells that may have a versatile and abundant application in regenerative medicine and dentistry due to their availability, possibility of isolation from different types of teeth and potential to differentiate into several cell types. In recent years, there is an emerging trend of the use of nanomaterials in medicine and dentistry that, by virtue of their unique properties, have become very attractive as a tool for the treatment of bone tissue defects. The aim of our study was to examine the potential of synthesized nanomaterial, intended for bone tissue engineering and regenerative applications, biphasic calcium phosphate coated with poly-D,L-lactide-co-glycolide (CP/PLGA), to influence the osteogenic differentiation of DPSCs. Cells were obtained from the mature healthy teeth by outgrowth of the cells from undigested pulp pieces during culturing, in standard cell culture conditions. Cells were subjected to osteogenic differentiation for seven and 14 days by culturing the cells with two concentrations of CP/PLGA nanoparticles in the presence or absence of osteogenic supplements in the media. Osteogenic differentiation was assessed by phase contrast microscopy as well as by Von Kossa and Alizarin Red S staining of formed inorganic deposits. The results showed that CP/PLGA influenced osteogenesis in concentration-dependent manner and differently in osteogenic and standard cell culture media. The use of calcium phosphate-based nanomaterials in combination with DPSCs, under certain conditions, could be a promising approach in regenerative medicine and dentistry