ROLE OF PHAGOCYTES IMMUNE DEFENCE IN ANIMAL ORGANISM

Imunološki sustav je sustav organa koji štiti organizam od napada stranih mikroorganizama. Razlikujemo nespecifičnu i specifičnu imunost. Nespecifična imunost djeluje bez prethodnog kontakta organizma sa antigenom. Nespecifična imunost se temelji na mehanizmima kao što su anatomske prepreke, fiziološke prepreke, sustav komplementa, interferon, lizozim, te stanični mehanizmi (fagociti). Neutrofili iz krvotoka migriraju kroz endotelne stanice, nakon čega se moraju aktivirati da bi započela fagocitoza. Tako se aktivirani neutrofili usmjereno kreću prema mikroorganizmima, što je posljedica djelovanja kemotatičnih molekula. Proces fagocitoze započinje ingestijom kod koje se mikroorganizam uvlači u neutrofil, azavršava digestijom kod koje se probava mikroorganizma uništava lizirajućim enzimima i antimikrobnim peptidima. Nakon djelovanja na mjestu upale slijedi apoptoza neutrofila. Apoptozu reguliraju brojni citokini. Nakon apoptoze, apoptotičke neutrofile uklanjaju stanice mononuklearnog fagocitnog sustava.The immune system is the system that protects the body against attacks by foreign microorganisms. There are two types of immunity: innate and adaptive immunity. Innate immunity acts without prior contact with the antigen of the organism. Innate immunity is based on a number of mechanisms such as anatomic barriers, physiological barriers, the complement system, interferon, lysozyme, and cellular mechanisms (phagocytes). The neutrophils migrate from the bloodstream through the endothelial cells, and they must be activated to start the phagocytosis. Activated neutrophils have directed movement to microorganisms as a result of the chemotactic molecules. The process of phagocytosis begins with ingestion where the microorganism is drawn into the neutrophil, and ends with digestion in which the microorganisms is destroyed by lysing enzymes and antimicrobial peptides. Apoptosis is regulated by numerous cytokines. After apoptosis, apoptotic neutrophils are removed by mononuclear phagocytic syste

Dokazivanje gena za enterotoksine koagulaza-negativnih stafilokoka iz autohtonih hrvatskih fermentiranih kobasica.

The aim of this study was to determine the presence of enterotoxin genes in coagulase - negative staphylococci from traditional Croatian fermented sausages. A total of 39 isolates were collected from the dry sausage “kulen” and Slavonian homemade sausages as presumptive CNS, and were subjected to PCR identification to the genus level. All isolates belonged to the Staphylococcus genus. Strains were tested for the presence of 13 enterotoxin genes; seA, seB, seC, seG, seI, tsst1, seD, seE, seH, seJ, seM, seN and seO by the PCR method. All strains were negative to all enterotoxin genes. The observed results indicate the absence of risk related to enterotoxogenic potential of coagulase - negative staphylococci as indigenous microbiota from autochthonous Croatian traditional sausages.Cilj ovog rada bio je utvrditi prisutnost gena za enterotoksine u koagulaza - negativnih stafilokoka iz tradicionalnih hrvatskih fermentiranih kobasica. Ukupno je prikupljeno 39 izolata iz kulena i domaće slavonske kobasice te identificirano PCR - om do razine roda. Svi izolati pripadali su rodu Staphylococcus. PCR - om je određivana prisutnost 13 gena za enterotoksine: seA, seB, seC, seG, seI, tsst1, seD, seE, seH, seJ, seM, seN i seO. Ni u jednom izolatu nije utvrđena prisutnost gena za enterotoksine. Dobiveni rezultati upućuju na odsutnost rizika povezanog s toksogenim potencijalom koagulaza-negativnih stafilokoka kao dijela autohtone mikroflore hrvatskih fermentiranih kobasica

Dokazivanje gena za enterotoksine koagulaza-negativnih stafilokoka iz autohtonih hrvatskih fermentiranih kobasica.

The aim of this study was to determine the presence of enterotoxin genes in coagulase - negative staphylococci from traditional Croatian fermented sausages. A total of 39 isolates were collected from the dry sausage “kulen” and Slavonian homemade sausages as presumptive CNS, and were subjected to PCR identification to the genus level. All isolates belonged to the Staphylococcus genus. Strains were tested for the presence of 13 enterotoxin genes; seA, seB, seC, seG, seI, tsst1, seD, seE, seH, seJ, seM, seN and seO by the PCR method. All strains were negative to all enterotoxin genes. The observed results indicate the absence of risk related to enterotoxogenic potential of coagulase - negative staphylococci as indigenous microbiota from autochthonous Croatian traditional sausages.Cilj ovog rada bio je utvrditi prisutnost gena za enterotoksine u koagulaza - negativnih stafilokoka iz tradicionalnih hrvatskih fermentiranih kobasica. Ukupno je prikupljeno 39 izolata iz kulena i domaće slavonske kobasice te identificirano PCR - om do razine roda. Svi izolati pripadali su rodu Staphylococcus. PCR - om je određivana prisutnost 13 gena za enterotoksine: seA, seB, seC, seG, seI, tsst1, seD, seE, seH, seJ, seM, seN i seO. Ni u jednom izolatu nije utvrđena prisutnost gena za enterotoksine. Dobiveni rezultati upućuju na odsutnost rizika povezanog s toksogenim potencijalom koagulaza-negativnih stafilokoka kao dijela autohtone mikroflore hrvatskih fermentiranih kobasica

Aging affects rat inflammatory peritoneal exudate composition and macrophage inflammatory mediator production in a strain-dependent manner

The present study was designed to examine influence of aging on macrophage proinflammatory/anti-inflammatory capacity in rat model of thioglycollate-induced peritonitis. Peritoneal macrophages were isolated from young (3-months-old) and aged (18-months-old) Dark Agouti (DA) and Albino Oxford (AO) rats seven days post-injection of thioglycollate medium. Freshly isolated peritoneal exudate cells were examined for the expression of CD163, CCR7, CD14 and TLR4, whereas cytokine production (TNF-α, IL-6 and IL-10) and arginine metabolism end-products (NO and urea) were assayed in vitro under basal conditions and following stimulation with LPS. In DA rat inflammatory peritoneal exudate, aging diminished the frequency of cells with a “resolving macrophage” CD14+CD163+ phenotype. However, in AO rats, which exhibited stable frequency of CD14+CD163+ cells in inflammatory peritoneal exudate with aging, the proportion of CCR7-bearing peritoneal cells, presumably immigrating inflammatory monocytes, was diminished in aged animals. Under basal culture conditions, macrophages from aged rats of both strains released less amount of TNF-α, IL-6 and IL-10, but produced more urea than cells from young strain-matched rats. However, these changes were more pronounced in peritoneal macrophages from AO rats. Additionally, age-related decrease in the frequency of TLR4-expressing cells was observed among fresh peritoneal exudate cells from AO rats. Upon LPS stimulation, the production of prototypic inflammatory cytokines (TNF-α and IL-6) was diminished in macrophages from aged AO rats, whereas aging had the opposite effect on their production in DA rat macrophages. Moreover, aging increased NO production in LPS-stimulated macrophages from DA rats, whereas urea production was enhanced in macrophages from both strains, but this increase was strikingly more pronounced in macrophages from AO rats. Collectively, results suggest that aging affects inflammatory peritoneal exudate cellular composition and macrophage proinflamatory/immunomodulatory capacity in a strain- specific manne

Analysis of the Salt Content in Bread and Bakery Products after the Implementation of New National Regulation on Cereals and Cereal Products

Prekomjerni unos soli ključni je čimbenik u epidemiji prehipertenzije/hipertenzije. Hrvatska je 2014. objavila Strateški plan za smanjenje prekomjernog unosa kuhinjske soli u Republici Hrvatskoj 2014. – 2019. prema preporukama Svjetske zdravstvene organizacije te je preporučena količina soli 5 grama na dan (2 g natrija), dok prema najnovijim istraživanjima stanovnici u Hrvatskoj i dalje prosječno dnevno unose dvostruko više od preporučene dnevne količine (11,6 g na dan). U eksperimentalnom dijelu ovog istraživanja u slučajno odabranim uzorcima iz maloprodaje analitički je ispitan sadržaj natrija (te računski ukupni sadržaj soli) u više vrsta pekarskih proizvoda (kruh, peciva). U tu svrhu, primjenom sofisticirane analitičke opreme ICP-MS (induktivno spregnuta plazma s masenom spektrometrijom) utvrđene su količine natrija u različitim vrstama kruha, kruščića i peciva te uspoređene sa zahtjevima nacionalne regulative o maksimalno dozvoljenom sadržaju natrija u kruhu koji propisuje maksimalno 1,4 %. Prosječna vrijednost svih uzoraka iznosi 1,46 g / 100 g, što prelazi dozvoljenu količinu. Rezultati ukazuju na to da se svi proizvođači još uvijek nisu prilagodili odredbama pravilnika te da ne zadovoljavaju zahtjeve premašujući dozvoljene količine natrija, odnosno soli u kruhu i pecivima.Excessive salt intake is a key factor in the epidemic of prehypertension/hypertension. In 2014 the Republic of Croatia has published a National Strategic Plan for Reducing Excessive Salt Intake in the Republic of Croatia 2014-2019, according to the recommendations of the World Health Organization. The recommended intake of salt is 5 grams/day (2 grams of sodium), while according to the latest research, the population in Croatia on average still consume twice the recommended daily amount (11.6 grams/day). In the experimental part of this study, the sodium content (and the calculated total salt content) in several types of bakery products (bread, pastry) was analysed in randomly selected retail samples. For this purpose, using sophisticated analytical equipment ICP-MS (Inductively Coupled Plasma Mass Spectrometry), the amount of sodium in different types of bread, bakery products and pastry has been determined and compared with the requirements of the national regulation on maximum permitted sodium content in bread, which prescribes a maximum of 1.4 %. The average value of all samples is 1.46 grams/100 grams, which exceeds the permitted amount. The results indicate that not all manufacturers have yet complied with the regulations and that they do not meet the requirements exceeding the permitted amounts of sodium, i.e. salt in bread and pastries

Aging diminishes the resistance of AO rats to EAE: putative role of enhanced generation of GM-CSF Expressing CD4+T cells in aged rats

Background: Aging influences immune response and susceptibility to EAE in a strain specific manner. The study was designed to examine influence of aging on EAE induction in Albino Oxford (AO) rats. Results: Differently from 3-month-old (young) rats, which were resistant to EAE induction, the majority of aged (24-26-month-old) rats developed mild chronic form of EAE. On 16th day post-immunization, when in aged rats the neurological deficit reached plateau, more mononuclear cells, including CD4+ T lymphocytes was retrieved from spinal cord of aged than young rats. The frequencies of IL-17+ and GM-CSF+ cells within spinal cord infiltrating CD4+ lymphocytes were greater in aged rats. To their increased frequency contributed the expansion of GM-CSF + IL-17 + IFN-gamma+ cells, which are highly pathogenic in mice. The expression of the cytokines (IL-1 beta and IL-23/p19) driving GM-CSF + IL-17 + IFN-gamma + cell differentiation in mice was also augmented in aged rat spinal cord mononuclear cells. Additionally, in aged rat spinal cord the expansion of GM-CSF + IL-17-IFN-gamma- CD4+ T lymphocytes was found. Consistently, the expression of mRNAs for IL-3, the cytokine exhibiting the same expression pattern as GM-CSF, and IL-7, the cytokine driving differentiation of GM-CSF + IL-17-IFN-gamma- CD4 + lymphocytes in mice, was upregulated in aged rat spinal cord mononuclear cells, and the tissue, respectively. This was in accordance with the enhanced generation of the brain antigen-specific GM-CSF+ CD4+ lymphocytes in aged rat draining lymph nodes, as suggested by (i) the higher frequency of GM-CSF+ cells (reflecting the expansion of IL-17-IFN-gamma- cells) within their CD4+ lymphocytes and (ii) the upregulated GM-CSF and IL-3 mRNA expression in fresh CD4+ lymphocytes and MBP-stimulated draining lymph node cells and IL-7 mRNA in lymph node tissue from aged rats. In agreement with the upregulated GM-CSF expression in aged rats, strikingly more CD11b + CD45(int) (activated microglia) and CD45(hi) (mainly proinflammatory dendritic cells and macrophages) cells was retrieved from aged than young rat spinal cord. Besides, expression of mRNA for SOCS1, a negative regulator of proinflammatory cytokine expression in innate immunity cells, was downregulated in aged rat spinal cord mononuclear cells. Conclusions: The study revealed that aging may overcome genetic resistance to EAE, and indicated the cellular and molecular mechanisms contributing to this phenomenon in AO rats

GM-CSF-Producing Th Cells in Rats Sensitive and Resistant to Experimental Autoimmune Encephalomyelitis

Given that granulocyte macrophage colony-stimulating factor (GM-CSF) is identified as the key factor to endow auto-reactive Th cells with the potential to induce neuroinflammation in experimental autoimmune encephalomyelitis (EAE) models, the frequency and phenotype of GM-CSF-producing (GM-CSF+) Th cells in draining lymph nodes (dLNs) and spinal cord (SC) of Albino Oxford (AO) and Dark Agouti (DA) rats immunized for EAE were examined. The generation of neuroantigen-specific GM-CSF+ Th lymphocytes was impaired in dLNs of AO rats (relatively resistant to EAE induction) compared with their DA counterparts (susceptible to EAE) reflecting impaired CD4+ lymphocyte proliferation and less supportive of GM-CSF+ Th cell differentiation dLN cytokine microenvironment. Immunophenotyping of GM-CSF+ Th cells showed their phenotypic heterogeneity in both strains and revealed lower frequency of IL-17+ IFN-gamma+, IL-17+ IFN-gamma-, and IL-17-IFN-gamma+ cells accompanied by higher frequency of IL-17-IFN-gamma- cells among them in AO than in DA rats. Compared with DA, in AO rats was also found (i) slightly lower surface density of CCR2 (drives accumulation of highly pathogenic GM-CSF+ IFN-gamma+ Th17 cells in SC) on GM-CSF+ IFN-gamma+ Th17 lymphocytes from dLNs, and (ii) diminished CCL2 mRNA expression in SC tissue, suggesting their impaired migration into the SC. Moreover, dLN and SC cytokine environments in AO rats were shown to be less supportive of GM-CSF+ IFN-gamma+ Th17 cell differentiation (judging by lower expression of mRNAs for IL-1 beta, IL-6 and IL-23/p19). In accordance with the (i) lower frequency of GM-CSF+ Th cells in dLNs and SC of AO rats and their lower GM-CSF production, and (ii) impaired CCL2 expression in the SC tissue, the proportion of proinflammatory monocytes among peripheral blood cells and their progeny (CD45(hi) cells) among the SC CD11b+ cells were reduced in AO compared with DA rats. Collectively, the results indicate that the strain specificities in efficacy of several mechanisms controlling (auto) reactive CD4+ lymphocyte expansion/differentiation into the cells with pathogenic phenotype and migration of the latter to the SC contribute to AO rat resistance to EAE

Sex Bias in Pathogenesis of Autoimmune Neuroinflammation: Relevance for Dimethyl Fumarate Immunomodulatory/Anti-oxidant Action

In the present study, upon showing sexual dimorphism in dimethyl fumarate (DMF) efficacy to moderate the clinical severity of experimental autoimmune encephalomyelitis (EAE) in Dark Agouti rats, cellular and molecular substrate of this dimorphism was explored. In rats of both sexes, DMF administration from the day of immunization attenuated EAE severity, but this effect was more prominent in males leading to loss of the sexual dimorphism observed in vehicle-administered controls. Consistently, in male rats, DMF was more efficient in diminishing the number of CD4+ T lymphocytes infiltrating spinal cord (SC) and their reactivation, the number of IL-17+ T lymphocytes and particularly cellularity of their highly pathogenic IFN-gamma+GM-CSF+IL-17+ subset. This was linked with changes in SC CD11b+CD45+TCR alpha beta- microglia/proinflammatory monocyte progeny, substantiated in a more prominent increase in the frequency of anti-inflammatory phygocyting CD163+ cells and the cells expressing high surface levels of immunoregulatory CD83 molecule (associated with apoptotic cells phagocytosis and implicated in downregulation of CD4+ T lymphocyte reactivation) among CD11b+CD45+TCR alpha beta- cells in male rat SC. These changes were associated with greater increase in the nuclear factor (erythroid-derived 2)-like 2 expression in male rats administered with DMF. In accordance with the previous findings, DMF diminished reactive nitrogen and oxygen species generation and consistently, SC level of advanced oxidation protein products, to the greater extent in male rats. Overall, our study indicates sex-specificity in the sensitivity of DMF cellular and molecular targets and encourages sex-based clinical research to define significance of sex for action of therapeutic agents moderating autoimmune neuroinflammation-/oxidative stress-related nervous tissue damage

Ružička days : International conference 16th Ružička Days “Today Science – Tomorrow Industry” : Proceedings

Proceedings contains articles presented at Conference divided into sections: open lecture (1), chemical analysis and synthesis (3), chemical and biochemical engineering (8), food technology and biotechnology (8), medical chemistry and pharmacy (3), environmental protection (11) and meeting of young chemists (2)

ROLE OF PHAGOCYTES IMMUNE DEFENCE IN ANIMAL ORGANISM

Imunološki sustav je sustav organa koji štiti organizam od napada stranih mikroorganizama. Razlikujemo nespecifičnu i specifičnu imunost. Nespecifična imunost djeluje bez prethodnog kontakta organizma sa antigenom. Nespecifična imunost se temelji na mehanizmima kao što su anatomske prepreke, fiziološke prepreke, sustav komplementa, interferon, lizozim, te stanični mehanizmi (fagociti). Neutrofili iz krvotoka migriraju kroz endotelne stanice, nakon čega se moraju aktivirati da bi započela fagocitoza. Tako se aktivirani neutrofili usmjereno kreću prema mikroorganizmima, što je posljedica djelovanja kemotatičnih molekula. Proces fagocitoze započinje ingestijom kod koje se mikroorganizam uvlači u neutrofil, azavršava digestijom kod koje se probava mikroorganizma uništava lizirajućim enzimima i antimikrobnim peptidima. Nakon djelovanja na mjestu upale slijedi apoptoza neutrofila. Apoptozu reguliraju brojni citokini. Nakon apoptoze, apoptotičke neutrofile uklanjaju stanice mononuklearnog fagocitnog sustava.The immune system is the system that protects the body against attacks by foreign microorganisms. There are two types of immunity: innate and adaptive immunity. Innate immunity acts without prior contact with the antigen of the organism. Innate immunity is based on a number of mechanisms such as anatomic barriers, physiological barriers, the complement system, interferon, lysozyme, and cellular mechanisms (phagocytes). The neutrophils migrate from the bloodstream through the endothelial cells, and they must be activated to start the phagocytosis. Activated neutrophils have directed movement to microorganisms as a result of the chemotactic molecules. The process of phagocytosis begins with ingestion where the microorganism is drawn into the neutrophil, and ends with digestion in which the microorganisms is destroyed by lysing enzymes and antimicrobial peptides. Apoptosis is regulated by numerous cytokines. After apoptosis, apoptotic neutrophils are removed by mononuclear phagocytic syste