Karakteristike tkivno specifičnih makrofaga pre i posle aktivacije in vitro

Macrophages derived from different tissues: bone marrow, spleen, peritoneal cavity and alveolus, were examined from the aspects of their morphology and functional characteristics expression of Fc receptors (FcR), phagocytic activity towards yeast particles and nonspecific esterase (NSE) content¹ before and after in vitro activation. Twenty four-hour-adherent cells were isolated with the aim of analyzing the characteristics of resident tissue macrophages. Following cultivation in vitro 8-day-adherent cells were used to investigate the influence of macrophage activation on their morphology and function. Morphological analysis of cell smears, performed in respect to cell size, showed significant enlargement, especially in the population of alveolar cells cultured for 8 days and activated with colony-stimulating factors (CSFs) and lymphokines. It was also demonstrated that 24-hour- and 8-dayadherent macrophages derived from different tissues exhibited similar properties. All these cells were more than 90% FcR-positive (FcR+) NSE-positive (NSE+) and had phagocytic properties. However, within the population of alveolar macrophages there were some NSE+ cells lacking FcR and phagocytic activity, even after in vitro activation. These results confirmed that the properties of alveolar macrophages differing from those of macrophages from other tissues were dependent on their microenvironment.Morfološke i funkcionalne karakteristike makrofaga izolovanih iz različitih organa: kostne srži, slezine, peritonealne šupljine i alveola ispitivane su pre i posle aktivacije u in vitro uslovima. Da bi se ispitale karakteristike makrofaga koji se nalaze u ispitivanim tkivima analizirane su adherentne ćelije dobijene nakon inkubacije od 24 časa. Uticaj aktivacije makrofaga na njihovu morfologiju i funkciju (ekspresiju Fc receptora, sposobnost fagocitoze čestica kvasca i sadržaj enzima nespecifične esteraze) ispitivan je nakon kultivisanja adherentnih ćelija u toku 8 dana. Morfološkom analizom utvrđeno je značajno povećanje veličine alveolarnih makrofaga kultivisanih tokom 8 dana u prisustvu faktora koji stimulišu rast kolonija i limfokina. Pokazano je da adherentne ćelije iz različitih tkiva izolovane nakon 24 sata i 8 dana imaju slične funkcionalne karakteristike. Više od 90% tih ćelija je eksprimiralo Fc receptore, imalo sposobnost fagocitoze i sadržavalo nespecifičnu esterazu. Međutim, u populaciji alveolarnih makrofaga, pre kao i nakon in vitro aktivacije, utvđeno je prisustvo ćelija koje su sadržavale nespecifičnu esterazu u citoplazmi, ali nisu eksprimirale Fc receptore, niti su imale sposobnost fagocitoze. Ovi rezultati potvrđuju da su karakteristike alveolarnih makrofaga u odnosu na makrofage iz drugih tkiva zavisne od njihovog mikrookruženja

Proliferacija naivnih i aktiviranih T limfocita u prisustvu tkivno specifičnih makrofaga

In this study the antigen-presenting ability of tissue specific macrophages isolated from bone marrow, spleen, peritoneal cavity and lungs was analyzed. Murine macrophages were isolated by a one-step adherence procedure (for 24 hours) and pretreated with mytomycin C. The antigen-presenting ability of the macrophages was tested in T cell proliferation assays. The ability of macrophages to support antigenspecific proliferation of T lymphoblasts was investigated when sheep red blood cell (SRBC)-specific T blasts were stimulated in vitro by antigen in the presence of different numbers of tissue specific macrophages. On the other hand, the abilities of macrophages to induce proliferation of naïve T cells were analyzed in allogeneic and syngeneic mixed leukocyte reactions (MLRs). It was demonstrated that tissue specific macrophages supported antigen specific proliferation of T lymphoblasts in vitro. They also induced the activation of allogeneic and syngeneic T cells. Increasing the number of macrophages co-cultured with T cells, led to a certain inhibitory effect on T cell proliferation.U ovom radu je ispitivana sposobnost mišjih makrofaga izolovanih iz kostne srži, slezine, peritonealne šupljine i alveola da prezentuju antigen i indukuju proliferaciju T limfocita. Makrofazi su izolovani adherencijom tokom 24-časovne kulture i pretretirani mitomicinom C. Da bi se ispitala sposobnost makrofaga da indukuju proliferaciju limfoblasta, T limfociti specifični za ovčje eritrocite izolovani iz limfnih čvorova imunizovanih miševa u in vitro uslovima, su restimulisani antigenom u prisustvu tkivnih makrofaga izolovanih iz različitih tkiva. Pored toga, sposobnost makrofaga da indukuju proliferaciju in vivo naivnih T limfocita ispitivana je u mešanoj kulturi tkivnih makrofaga i alogenih ili singenih limfocita. Dokazano je da tkivni makrofazi izolovani iz kostne srži, slezine, peritonealne šupljine i alveola mogu da potpomognu antigen-specifičnu proliferaciju T limfoblasta u in vitro uslovima, kao i da aktiviraju naivne alogene i singene T limfocite. Povećanje broja makrofaga dovelo je do smanjenja T limfocitne proliferacije

Enhancing the customer relationship management in public libraries: findings from three developing countries

Purpose – The purpose of this paper is to investigate the possibilities of implementing the customer relationship management (CRM) concept and smart technologies in public libraries in developing countries. The goal was to explore the level of librarians’ awareness about CRM concept and their willingness to accept the CRM concept in libraries. Also, patrons’ satisfaction with the quality of services and relationships in public libraries is explored. Design/methodology/approach – The authors obtained data for this research through two online surveys. The first survey measures librarians’ level of awareness about CRM concept and their perception about CRM and smart technologies concept in public libraries in three developing countries: Serbia, Montenegro and Bosnia and Herzegovina. The second survey measures patrons’ satisfaction with services and relationship in public libraries in these three countries. Findings – The research results revealed that most surveyed librarians are familiar with the CRM concept. However, libraries in these three developing countries barely use CRM or smart technologies to improve the relationship with stakeholders. Also, most patrons are satisfied with relationships and services. The analysis of data indicates no significant difference in the satisfaction level among patrons between these three developing countries. Research limitations/implications – Not all libraries participated in this research. Therefore, the sample is not distributed evenly across different types of libraries or user groups. Future research should include different types of libraries. Practical implications – The results can be used by public libraries in developing countries to improve the relationship with stakeholders harnessing CRM concept and smart technologies. Social implications – The use of CRM in combination with the smart technologies can help leverage the quality of the relationship between public libraries and stakeholders which in turn would secure their support and loyalty in the future. Originality/value – The integration of CRM concept as a component of library business automation process is an idea that has not been discussed widely in the library community and could initiate a positive trend in public libraries in developing countrie

THE IMPORTANCE OF CASH FLOW AS QUALITY INDICATOR OF ENTREPRENEURIAL SUCCESS

U bilježenju poslovnih transakcija primjenjuju se dva ključna računovodstvena principa. Prema principu novčanog toka, prihodi, rashodi i ostale transakcije se ne priznaju dok no¬vac nije primljen ili plaćen. Prema obračunskoj metodi (obračunskoj osnovi), koji je danas opće prihvaćeni računovodstveni princip, performanse i pozicija poduzeća mjere se prizna¬vanjem ekonomskih događaja bez obzira na novčanu realizaciju transakcije. Obračunska računovodstvena osnova stoga omogućuje poduzećima kombiniranje informacija o teku¬ćim priljevima i odljevima s budućim očekivanim novčanim priljevima i odljevima u cilju stjecanja pouzdanije slike o tekućem financijskom položaju poduzeća. Obračunsko raču¬novodstvo i računovodstveni standardi poduzećima dozvoljavaju do određene mjere izbor (između različitih opcija) u računovodstvenom tretmanu poslovnih transakcija. Ponekad bilježenje računovodstvenih kategorija ne odražava istinsku novčanu supstancu transak¬cije, omogućujući prezentiranje veće ili manje dobiti u odnosu na ekonomsku stvarnost. Prepoznajući ovu mogućnost za poduzeća, investitori su u posljednjih desetak godina sve više pažnje posvećivali izvještaju o novčanom toku kao vrijednom izvoru u ocjeni kvalitete dobiti. Iako je ovo razboriti korak, investitori moraju biti posebno oprezni. Menadžment je implementirao kreativne načine umjetnog povećanja novčanog toka iz operativnih aktiv¬nosti. Novčani tok iz operativnih aktivnosti je ključni pokazatelj na koji se oslanjaju investi¬tori u procjeni poduzeća da kreira novac, kao i njegovu kvalitetu dobiti. Članak osvjetljuje važnost izvještaja o novčanom toku i razmatra određene probleme pri klasifikaciji u kreira¬nju izvještaja o novčanom toku.There are two key accounting principles in recording business transactions. Under the cash flow principle, revenues, expenses and other transactions are not recognized until cash is actually received or paid. Under the accrual method (accrual basis), which is nowadays widely accepted accounting principle, performance and position of a company is measu¬red by recognizing economic events regardless of when cash transactions occur. Accrual basis of accounting thus allows the current cash inflows and outflows information to be combined with information about future expected cash inflows and outflows in order to acquire a more reliable picture of a company’s current financial position. Accrual accoun¬ting and accounting standards allow to certain degree for companies to choose (among available options) accounting treatment of transactions. Thus certain accounting records may not reflect true economic substance of transactions allowing earnings to be presented in opposite to its economic reality. Recognizing this opportunity for companies, in the last decade investors have increasingly turned their attention to cash flow statement as a valu¬able source in evaluating quality of earnings. While this can be a wise step, investors must be particularly careful. Management employed creative ways as well to artificially inflate reported cash flow from operations. Cash flow from operations is a key indicator investors rely upon to assess both a company’s ability to generate cash and its quality of earnings. This article enlightens importance of cash flow statement and discusses certain classifica¬tion issues in creating cash flow statement

Monoklonsko antitelo 26 napravljeno na tetanus toksoidu reaguje i sa tetanus toksinom i β2-glikoproteinom I - karakteristike vezivanja in vitro i moguća primena

A murine monoclonal IgG1 antibody, marked as MAb26, specific for tetanus toxoid has been immunochemically characterized. By performing enzyme-linked immunosorbent assays (ELISAs) and western blot analyses, it was demonstrated that MAb26 reacted with tetanus toxoid, tetanus toxin and β2-glycoprotein I (β2GPI). According to the results, MAb26 recognized the sequential epitope on the tetanus heavy chain. The affinity constant, calculated from Scatchard plots of MAb26 binding to tetanus toxoid, was 1.145×108 M-1 and the measurement of the relative affinity of MAb26 by ELISA using thiocyanate elution showed a significantly higher affinity of MAb26 to the toxoid (p = 0.0012) in comparison to the toxin. Additionally, the reactivity of MAb26 toward the toxoid forms increased when the tetanus toxin was detoxified using 8 mM and higher formaldehyde concentrations. The similarity of the tetanus toxoid to several sera proteins, either at the level of its conformation (IL-1α) or at the level of peptide sequences (â2GPI, laminin) favors its role in autoimmunity by the mechanism of molecular mimicry. As the induction of an autoimmune disease is dependent on the breakdown of tolerance, which could be the result of an overt hyperstimulation, the control of the presence and concentration of self-reactive epitopes in vaccine preparations is a prerequisite. In this study, it was shown that MAb26 can: 1) discriminate between the tetanus toxin and different toxoid forms, which makes it a good candidate for antibody control during vaccine preparation; 2) due to its cross-reactivity with β2GPI, it could provide information on the presence of a potentially dangerous sequential epitope expressed at the protein surface.Ovaj rad opisuje imunohemijsku karakterizaciju mišjeg IgG1 monoklonskog antitela označenog kao MAt26. Enzimskim imunosorbentnim testom (ELISA) i Western blot analizom je pokazano da MAt26 reaguje sa tetanus toksoidom, tetanus toksinom i β2-glikoproteinom I (β2GPI). Prema našim rezultatima, MAt26 prepoznaje sekvencioni epitop na teškom lancu molekula tetanusa. Konstanta afiniteta MAt26 za tetanus toksoid, izračunata na osnovu Skačardovog dijagrama, je 1,145×108 M-1. Na osnovu elucije tiocijanatom, korišćene za određivanje relativnog afiniteta MAt26 za tetanus toksin i tetanus toksoid, postupkom baziranim na ELISA-i, pokazan je znatno veći (p = 0,0012) afinitet MAt26 ka toksoidnoj formi. Takođe, reaktivnost MAt26 ka toksoidnoj formi rasla je sa porastom koncentracije formaldehida, počevši od 8 mM, korišćenog u procesu detoksifikacije. Sličnost tetanus toksoida sa različitim serumskim proteinima na nivou konformacije i/ili peptidnih sekvencija (β2GPI, laminin) ukazuje na njegovu potencijalnu ulogu u indukciji autoimunosti mehanizmom molekulske mimikrije. Budući da nastanak autoimunske bolesti podrazumeva narušavanje tolerancije, na primer, prekomernom stimulacijom imunskog sistema, kontrola prisustva i koncentracije sebi sličnih epitopa se nameće kao neophodna. U ovom radu je pokazano da: 1) MAt26 može da pravi razliku između tetanus toksina i različitih toksoidnih formi što ga čini potencijalno dobrim antitelom koje bi se koristilo u kontroli tokom proizvodnje vakcina; 2) zahvaljujući unakrsnoj reaktivnosti sa β2GPI, MAt26 može da pruži informacije o prisustvu potencijalno opasnih epitopa na površini proteina

The Possible Role of Natural Idiotopes in Immune Memory

In this paper we report on the generation of Abs possessing specificities similar to those of Abs used in immunization, and on the generation of Id and anti-Id specificities in the sera of mice immunized with commensal bacterial antigens

Network connectivity is shown to change in C57BL/6 mice during a continuing immune response subsequent to tetanus toxoid hyperimmunization

We have already demonstrated (Stojanovic et al., 2009) a connection between tetanus toxoid (TTd) hyperimmunization and the induction of anti-phospholipid syndrome (APS) in BALB/c mice. Here we show that C57BL/6 mice subjected to an identical procedure do not exhibit any like pathology attributable to anti-phospholipid antibodies; we explain that this absence results from idiotypic connectivity. Six groups of C57BL/6 mice were hyperimmunized with TTd in aluminum hydroxide or glycerol, with or without pretreatments. Pretreated mice had been injected with polyclonal or nonspecific immune stimulators, such as complete Freund's adjuvant (CFA) or glycerol. The epitope specificity of induced antibodies was tested by indirect ELISA using a tetanus toxoid immunogen and these autoantigens: phospholipids, gangliosides, laminin. Idiotypic connectivity was tested by competitive ELISA and gauged from the degree to which the interaction of idiotypic/anti-idiotypic complementary antibodies was inhibited in the presence of immunized sera antibodies. Higher idiotypic connectivity was noted amongst pretreated mice. There was a positive correlation between higher connectivity and autoantibody levels that acted to favor the participation of natural autoantibodies in the inhibitory process. We conclude that idiotypic connectivity plays a protective role in immunization-induced autoimmunity

Efekti IL-17 na funkcionalnu aktivnost ćelija periferne krvi

Interleukin-17 (IL-17) is a proinflammatory cytokine produced mainly by activated CD4+ and CD8+ T cells, while its specific receptor is ubiquitously distributed. The inflammatory capacity of IL-17 is based on its ability to stimulate a wide range of stromal cells to produce and release a number of proinflammatory mediators, some with a known impact on hematopoiesis particularly granulopoiesis. Recent data indicate a role for IL-17 in the pathogenesis of several inflammatory diseases, transplant rejection and tumor growth. The purpose of this study was to determine functional responses including the respiratory burst, nitric oxide (NO) production, adhesiveness and metabolical activity/viability of human peripheral blood leukocytes (total white blood cells, mononuclear cells and granulocytes) from healthy donors in the presence of recombinant human (rh)IL-17. The obtained results showed that rhIL-17 did not induce significant changes in the respiratory burst, NO production, and metabolical activity of each peripheral blood cell fraction the tested, while a slight increase in phorbol-12-myristate-13-acetate (PMA) stimulated adhesiveness of granulocytes and mononuclear cells was noted. The absence of significant changes in tested functional activities of various peripheral blood cells suggests that IL-17 does not express its proinflammatory ability in steady-state, since the requirement for its action really does not exist.Interleukin 17 (IL-17) je proinflamatorni citokin koga produkuju aktivirane CD4+ i CD8+ T ćelije, dok je njegov receptor ubikvitarno distribuiran. Inflamatorni kapacitet IL-17 se zasniva na njegovoj sposobnosti da stimuliše širok spektar stromalnih ćelija da produkuju i oslobađaju različite proinflamatorne medijatore, među kojima neki imaju efekte na hematopoezu posebno granulopoezu. Dosadašnji podaci ukazuju na ulogu IL-17 u patogenezi različitih inflamatornih bolesti, odbacivanju transplanta i razvoju tumora. Cilj ovog rada je bio da se odrede funkcionalni odgovori, uključujući respiratorni prasak, produkciju azot monoksida (NO), adhezivnost i metaboličku aktivnost/vijabilnost različitih ćelija periferne krvi (ukupnih leukocita, mononuklearnih ćelija i granulocita) zdravih donora, u prisustvu IL-17. Dobijeni rezultati su ukazali da IL-17 ne dovodi do značajnih promena respiratornog praska, produkcije NO i metaboličke aktivnosti ćelija periferne krvi, ali da uzrokuje blago povećanje forbol-12-miristat-13-acetat (PMA) stimulisane adhezivnosti granulocita i mononuklearnih ćelija. Odsustvo značajnih promena u ispitivanim funkcionalnim aktivnostima različitih ćelija periferne krvi, ukazuje da IL-17 ne eksprimira proinflamatorno dejstvo kod zdravih osoba, jer najverovatnije i ne postoji potreba za njegovim delovanjem

Odnos između totalnog kapaciteta za vezivanje gvožđa i koncentracije transferina kod novorođene prasadi tretirane gvožđe-dekstranom

Serum iron concentration and iron saturation of transferrin (Trf) are measures of body iron stores after administration of iron supplements. In clinical and experimental research, the complex determination of Trf was replaced by the simple determination of total iron binding capacity (TIBC). The objective of this work was to define if TIBC could be an adequate measure for Trf in neonatal piglets after i.m. iron administration. Treated piglets received 150 mg of iron-dextran i.m. the first day of life, and were compared to the untreated control group. Prior to iron administration, as well as on days 2, 8 and 12 after iron administration, serum iron and TIBC concentration were analyzed by an automatized chemical analyzer and Trf was determined by densitometry of electrophoretic strips. Our results show that regardless of iron treatment, TIBC is not a measure of Trf concentration in neonatal piglets two days after birth. At day 8 of their life a high correlation coefficient of these two parameters was established in non-treated animals, while in iron-treated piglets the same correlation was established 12 days after iron treatment. Thus, we suggest that in neonatal piglets, TIBC could be used as a measure of Trf concentration only 12 days after i.m. iron treatment.Određivanje statusa gvožđa u organizmu jedinke posle primene određenog preparata ovog mikroelementa moguće je utvrditi određivanjem njegove koncentracije u serumu i zasićenja transferina (Trf) gvožđem. U kliničkoj i eksperimentalnoj praksi složeno određivanje koncentracije Trf zamenjeno je jednostavnim određ ivanjem ukupnog serumskog kapaciteta za vezivanje gvožđa (TIBC). Cilj ovog rada je bio da se na modelu porasta serumskog Fe po i.m. aplikaciji Fe-dextrana novorođenoj prasadi, utvrdi odnos TIBC i Trf po aplikaciji ovog mikroelementa, kako bi se utvrdilo da li visoke doze gvožđa u serumu utiču na vrednost TIBC kao mere za određivanje koncentracije Trf. Vrednosti za serumsko Fe, TIBC i Trf poređene su između grupe životinja koja je odmah po rođenju dobila 150 mg Fe-dextrana i kontrolne grupe u kojoj životoinje nisu tretirane Fe-dextranom. Krv je uzorkovana pre aplikacije Fe-dextrana, drugog, osmog i dvanaestog dana po aplikaciji preparata gvožđa. Koncentracija gvožđa u serumu i TIBC su određivani standarnim kliničkim biohemijskim analizama, dok je koncenracija Trf određena denzitometrijom elektroforetskih traka. Dobijeni rezultati ukazuju da bez obzira na primenu preparata Fe, TIBC nije adekvatna mera za Trf kod novorođene prasadi u prva tri dana po rođenju. Osmog dana života prasadi, utvrđen je visoki stepen korelacije ova dva parametra kod životinja koje nisu bile tretirane, dok je kod tretiranih jedinki taj stepen korelacije postignut dvanaestog dana. Na osnovu izloženih rezultata se može zaključiti da se kod novorođene prasadi TIBC može koristiti kao mera za Trf tek 12 dana nakon i.m. tretmana Fe-dextranom