Praćenje in vitro biousvojivosti i preuzimanja glikozilovanih alergena hrane upotrebom ćelijskih modela

extensive engagement of researchers in the elucidation of the mechanisms underlying processes of food digestion, allergen transport, and uptake by the immune cells and its effector immune responses. The development of the in vitro assays and cell-based models has allowed bridging the problem of food allergy research and the respect of the ethical norms in used research procedures. The red meat allergy is a novel type of food allergy characterized by the production of an IgE antibody against the carbohydrate galactose-α-1,3-galactose (α- Gal). Glycoproteins from non-primate mammals are rich with an α-Gal as posttranslational modification. Also, red meat allergy is characterized by the delayed onset of symptoms which may be related to the mechanism and the fate of α-Gal carrying proteins in the human gastrointestinal tract. Furthermore, the uptake, processing, and mechanisms of presentation of α-Gal by the immune cells are still unknown. Therefore this doctoral dissertation aimed to investigate how protein glycosylation by α-Gal affects their susceptibility to gastric digestion, does α-Gal conjugated to proteins affects their transport through the Caco-2 cell monolayer, which mimics the gastrointestinal layer, and to examine the influence of α-Gal epitopes on the protein surface on their uptake and processing by immature monocyte-derived dendritic cells (iMDDCs). The study revealed that the presence of the α-Gal glycosylation on protein surface had an impact on their susceptibility to gastric digestion and the digestion pattern of the obtained protein fragments upon pepsinolysis. Prolonged survival, up to 2h of digestion, was characteristic of the large proteins fragments bearing the α-Gal epitope. Importantly, transport through the Caco-2 monolayer of proteins conjugated to α-Gal was hampered in comparison to unconjugated proteins. Furthermore, differential centrifugation of Caco-2 cell lysates upon transport experiments revealed that α-Gal could be detected on the intact protein in the endosomal fraction of the cells...Alergije na hranu su rastući problem u ljudskoj populaciji širom sveta i rešavanje ovog problema zahteva opsežno angažovanje istraživača u rasvetljavanju mehanizama uključenih u procese varenja hrane, transporta alergena i njihovog unosa od strane imunih ćelija odgovornih za efektorske mehanizme imunoloških odgovora. Ovo je nezamislivo bez razvoja in vitro testova i ćelijskih modela koji premošćuju problem istraživanja alergija na hranu uz poštovanja etičkih normi u korišćenim istraživačkim metodama. Novu vrstu alergije na hranu, alergiju na crveno meso, karakteriše sinteza imunoglobulina E kao odgovor na prisustvo šećera galaktoza-α-1,3-galaktoza (α-Gal), koji je prisutan na površini glikoproteina primata. Takođe, alergiju na crveno meso karakteriše odložena pojava simptoma što može biti rezultat promena u mehanizmu obrade proteina koji nose α-Gal u gastrointestinalnom traktu čoveka. Dalje, unos, obrada i mehanizmi prezentacije α-Gal šećera od strane imunih ćelija još uvek nisu poznati. Stoga ciljevi ove doktorske disertacije su ispitivanje kako α-Gal glikozilacija proteina utiče na njihovu digestiju od strane pepsina, da li α-Gal glikozilacija proteina utiče na njihov transport kroz monosloj Caco-2 ćelija, koji oponaša gastrointestinalni epitel, kao i ispitivanje uticaja α-Gal glikozilacije na površini proteina na njihov unos i obradu od strane nezrelih dendritičnih ćelijama kultivisanih iz monocita (iMDDC). Iz dobijenih rezultata moze se zaključiti da prisustvo α-Gal glikozilacije na površini proteina utiče na njihovu podložnost na digestiju a najviše na obrazac dobijenih fragmenata proteina nakon pepsinolize. Veliki fragmenti proteina koji nose α-Gal prisutni su čak i do 2 sata digestije. Takođe, važno je istaći da je transport α-Gal glikozilovanih proteina kroz Caco-2 monosloj otežan u poređenju sa neglikozilovanim proteinima. Dalje, diferencijalnim centrifugiranjem lizata aco-2 ćelija nakon transcitoze pokazalo je da je α-Gal prisutan na intaktnim proteinima u endozomalnim frakcijama ćelija..

Radiation-Induced method for the controlled synthesis of nanostructured materials

The synthesis represents a crucial step for achieving the unique properties of nanomaterials. Gamma (γ) - radiation-induced synthesis possesses several advantages over other conventional synthetic routes such as production of clean materials, the possibility of simultaneous synthesis and sterilization, in-situ generation of radiolysis products and the lack of necessity for initiators and crosslinking reagents in the case of polymer-based materials fabrication. γ-radiation induced synthesis has been utilized in the development and modification of various types of systems, including metal, metal oxide and alloyed nanoparticles, carbon-based nanomaterials, polymer based nanomaterials and nanocomposites etc. In this lecture, fundamentals of radiation chemistry will be given with an emphasis on the synthesis of nanomaterials in aqueous solutions. The effects of the total dose, dose rate, and the addition of different solutes to tailoring the synthesized materials’ composition, form, shape, size and size distribution will be demonstrated. The possible applications of so obtained materials in industry and biomedicine will be presented

Morfološke, imunofenotipske i kliničke karakteristike limfocitno - plazmocitnog duodenitisa pasa

The most important morphological feature of the immune response in lymphocytic-plasmacytic duodenitis (LPD) in dogs is the presence of a mononuclear infiltrate in the duodenal mucosa. The ethiopatogenesis of this disease is still unknown, nor are known all the immunophenotypic characteristics of the infiltrate cells, which would be of help in the elucidation of the pathogenesis of LPD. The study involved 60 adult dogs of different breeds and both sexes: 54 dogs with symptoms of diarrhea and vomiting that lasted longer than three weeks and six clinically healthy dogs that served as controls. Hematological and biochemical analysis of blood, radiology, ultrasound and endoscopic diagnosis were carried out. Samples of duodenal mucosal biopta were examined by histopathological and immunohistochemical methods. During endoscopic examination, duodenal wall thickening, mucosal edema and hyperemia were observed in 46 dogs. Microscopic studies showed on the epithelial cells of the duodenum degenerative changes, and sometimes desquamation. Most of the intestinal crypts were dilated, irregularly shaped and filled with detritus. On immunohistochemical examination of samples of the duodenum in a number of dogs with LPD a predominant expression of CD3 + T lymphocytes was observed.Najvažnija morfološka karakteristika imunskog odgovora kod limfocitnoplazmocitnog duodenitisa (LPD) pasa je nalaz infiltrata mononuklearnih ćelija krvi u sluzokoži duodenuma. Do danas etiopatogeneza ove bolesti pasa nije u svemu poznata, a nisu poznate ni sve imunofenotipske karakteristike ćelija u infiltratu, koje bi bile značajne za razumevanje etiopatogeneze LPD. Ispitivanjem je bilo obuhvaćeno ukupno 60 odraslih pasa, različitih rasa i oba pola: 54 psa sa simptomima dijareje i povraćanja koje je trajalo duže od tri nedelje i šest klinički zdravih pasa koji su služili kao kontrola. Urađene su hematološke i biohemijske analize krvi, a zatim je sprovedena radiološka, ultrazvučna i endoskopska dijagnostika. Uzorci mukoze duodenuma, uzeti biopsijom, ispitani su histopatološkim i imunohistohemijskim metodama. Pri endoskopskom pregledu, kod 46 pasa je zapažen zadebljali zid duodenuma, hiperemičnost i edem sluznice. Mikroskopskim ispitivanjima, na epitelnim ćelijama duodenuma uočene su degenerativne promene, a nekada i deskvamacija. Većina crevnih kripti je bila dilatirana, nepravilnog oblika i ispunjena detritusom. Imunohistohemijskim ispitivanjem uzoraka duodenuma kod većine pasa sa LPD uočena je dominacija CD3+T limfocita

Fatty acids composition of the most common bivalves in Korean diet

Consumption of bivalve molluscs, such as oysters, mussels, clams and scallops, makes a significant part of the daily Korean diet. Bivalves provide high quality proteins with all the dietary-essential amino acids, lipids, vitamins, minerals and other bioactive nutrients, which offer a variety of health benefits to the consumer [1]. This food contains less than 5 percent of total fat, so it is considered a low-fat food. Beside the amount of total fat, the proportions of saturated, monounsaturated and polyunsaturated fatty acids (FA) (S, M and P, respectively), as well as ratio of n-3 (ω-3) and n-6 (ω-6) P in food are very important for the health diet [2]. Fourteen species of bivalves Anadara broughtonii (AB), Ruditapes philippinarum (Manila clam (RP)), Tegillarca granosa (TG), Pecten yessoensis (Yesso scallop (YS), Argopecten spp. (small scallop (SS), Chlamys farreri farreri (CF), Cyclina sinensis (CS), Leukoma jedoensis (LJ), Mytilus califorianus (MCa) Mytilus galloprovancialis (MG), Maretrix lusoria (ML), Mactra quadrangularis (MQ), Sinovacula constricta (SC) and Crassostrea gigas (Pacific oyster (PC)) were bought in two fish markets in Incheon, Korea, in order to determine FA composition using GC/EI-MS of fatty acid methyl esters (FAME). The FAME were identified by comparing their retention times with those of the FAME standards or by comparing their mass spectra with those stored in the NIST Mass Spectral Library. In the bivalve samples, 43 different FA were identified, of which 10 were S, 12 M and 13 P, other FA were 7 methyl-FA and 1 hydroxyl-FA. The P/S ratio and ω-6/ω-3 P ratio are the most significant markers of lipid composition in a healthy diet and both should be close to 1 [3]. Among analysed species, only YS and SS have P/S ratio close to 1 (1,20 and 1.16, respectively), while other species have value between 0.07 and 0.73. The obtained values for ω-6/ω-3 P ratio were from 0.008 to 0.55, which indicates that bivalve molluscs are the valuable source of ω-3 P (EPA and DHA). These ω-3 P play important roles in growth, development, and maintenance of health

Fatty acids composition of the most common bivalves in Korean diet

Consumption of bivalve molluscs, such as oysters, mussels, clams and scallops, makes a significant part of the daily Korean diet. Bivalves provide high quality proteins with all the dietary-essential amino acids, lipids, vitamins, minerals and other bioactive nutrients, which offer a variety of health benefits to the consumer [1]. This food contains less than 5 percent of total fat, so it is considered a low-fat food. Beside the amount of total fat, the proportions of saturated, monounsaturated and polyunsaturated fatty acids (FA) (S, M and P, respectively), as well as ratio of n-3 (ω-3) and n-6 (ω-6) P in food are very important for the health diet [2]. Fourteen species of bivalves Anadara broughtonii (AB), Ruditapes philippinarum (Manila clam (RP)), Tegillarca granosa (TG), Pecten yessoensis (Yesso scallop (YS), Argopecten spp. (small scallop (SS), Chlamys farreri farreri (CF), Cyclina sinensis (CS), Leukoma jedoensis (LJ), Mytilus califorianus (MCa) Mytilus galloprovancialis (MG), Maretrix lusoria (ML), Mactra quadrangularis (MQ), Sinovacula constricta (SC) and Crassostrea gigas (Pacific oyster (PC)) were bought in two fish markets in Incheon, Korea, in order to determine FA composition using GC/EI-MS of fatty acid methyl esters (FAME). The FAME were identified by comparing their retention times with those of the FAME standards or by comparing their mass spectra with those stored in the NIST Mass Spectral Library. In the bivalve samples, 43 different FA were identified, of which 10 were S, 12 M and 13 P, other FA were 7 methyl-FA and 1 hydroxyl-FA. The P/S ratio and ω-6/ω-3 P ratio are the most significant markers of lipid composition in a healthy diet and both should be close to 1 [3]. Among analysed species, only YS and SS have P/S ratio close to 1 (1,20 and 1.16, respectively), while other species have value between 0.07 and 0.73. The obtained values for ω-6/ω-3 P ratio were from 0.008 to 0.55, which indicates that bivalve molluscs are the valuable source of ω-3 P (EPA and DHA). These ω-3 P play important roles in growth, development, and maintenance of health

Deficits in social cognition in adults with temporal lobe epilepsy

Interesovanje za specifičnosti socijalne kognicije osoba sa epilepsijom postaje aktuelnije kako zbog višestrukog uticaja koji ovaj poremećaj ima na psihosocijalnu adaptaciju i kvalitet života pogođene osobe, uprkos sve naprednijem medicinskom tretmanu, tako i zbog toga što sama epilepsija predstavlja pogodnu paradigmu za rasvetljavanje posebnih kognitivnih procesa koji se nalaze u pozadini socijalnog ponašanja. U radu je dat sažet prikaz istraživanja socijalne kognicije kod osoba sa ubedljivo najčešćom formom epilepsije kod odraslih – epilepsijom temporalnog režnja (temporalna epilepsija, TE), sa fokusom na komponentama ove funkcije koje danas privlače najveću pažnju. Uopšteno, pacijenti sa TE ispoljavaju poteškoće mentalizacije postižući slabije rezultate na različitim za- dacima (afektivne i kognitivne) teorije uma. Zatim, za ovu populaciju su ka- rakteristični deficiti prepoznavanja emocija, naročito negativnih, kao i slabije razvijena kognitivna komponenta empatije. Nalazi koji u celini sugerišu veći ili manji stepen narušenosti socijalne kognicije kod osoba sa TE mogu doprineti dubljem razumevanju socioemocional- nih poteškoća sa kojima se one suočavaju u svakodnevnom životu. U tom smis- lu, istraživanja socijalne kognicije daju vetar u jedra razvoju i poboljšanju dosadašnjih praksi lečenja epilepsije, ukazujući na neophodnost snažnijeg na- glaska na psihosocijalnim problemima, podjednako u dijagnostici kao i tret- manu, da bi se obezbedio holistički, sveobuhvatan tretman i zbrinjavanje pacijenata.There is a growing interest in the characteristics of social cognition in people with epilepsy, due to the multiple effects that epilepsy has on the psychosocial adaptation and quality of life of the patient, despite increasingly advanced medical treatment, and because epilepsy itself represents a suitable paradigm for elucidation of the cognitive processes underlying social behavior. The paper provides a brief overview of research on social cognition in people with temporal lobe epilepsy (TLE), the most prevalent type of epilepsy found in adults, with a focus on the components of this function that attract the most attention in the literature. In general, patients show deficits in various (affective and cognitive) Theory of Mind tasks. Emotion recognition deficits are common in patients with TLE and widespread across negative emotions, as well as deficits in cognitive empathy. Findings that overall suggest a certain degree of impairment of social cognition in patients with TLE can contribute to a deeper understanding of the socio-emotional difficulties they face in everyday life. In this sense, research on social cognition gives rise to the development and improvement of current epilepsy treatment practices, indicating the necessity of a stronger emphasis on psychosocial problems, in both diagnosis and treatment, in order to ensure holistic, comprehensive patient care

Hydrogen evolution reaction on rhenium modified phosphate Mo- and phosphate W-bronze

Phosphate molybdenum bronze (MoPB) and phosphate tungsten bronze (WPB) were synthesized and used as support for rhenium. Rhenium-modified bronze samples were applied at the surface of the glassy carbon electrode and tested for hydrogen evolution reaction. The electrodes were investigated by cyclic voltammetry and linear sweep voltammetry. The influence of carbon black on the electrochemical activity of starting bronze and Re-modified bronze samples was tested

Deficits in social cognition in adults with temporal lobe epilepsy

Interesovanje za specifičnosti socijalne kognicije osoba sa epilepsijom postaje aktuelnije kako zbog višestrukog uticaja koji ovaj poremećaj ima na psihosocijalnu adaptaciju i kvalitet života pogođene osobe, uprkos sve naprednijem medicinskom tretmanu, tako i zbog toga što sama epilepsija predstavlja pogodnu paradigmu za rasvetljavanje posebnih kognitivnih procesa koji se nalaze u pozadini socijalnog ponašanja. U radu je dat sažet prikaz istraživanja socijalne kognicije kod osoba sa ubedljivo najčešćom formom epilepsije kod odraslih – epilepsijom temporalnog režnja (temporalna epilepsija, TE), sa fokusom na komponentama ove funkcije koje danas privlače najveću pažnju. Uopšteno, pacijenti sa TE ispoljavaju poteškoće mentalizacije postižući slabije rezultate na različitim za- dacima (afektivne i kognitivne) teorije uma. Zatim, za ovu populaciju su ka- rakteristični deficiti prepoznavanja emocija, naročito negativnih, kao i slabije razvijena kognitivna komponenta empatije. Nalazi koji u celini sugerišu veći ili manji stepen narušenosti socijalne kognicije kod osoba sa TE mogu doprineti dubljem razumevanju socioemocional- nih poteškoća sa kojima se one suočavaju u svakodnevnom životu. U tom smis- lu, istraživanja socijalne kognicije daju vetar u jedra razvoju i poboljšanju dosadašnjih praksi lečenja epilepsije, ukazujući na neophodnost snažnijeg na- glaska na psihosocijalnim problemima, podjednako u dijagnostici kao i tret- manu, da bi se obezbedio holistički, sveobuhvatan tretman i zbrinjavanje pacijenata.There is a growing interest in the characteristics of social cognition in people with epilepsy, due to the multiple effects that epilepsy has on the psychosocial adaptation and quality of life of the patient, despite increasingly advanced medical treatment, and because epilepsy itself represents a suitable paradigm for elucidation of the cognitive processes underlying social behavior. The paper provides a brief overview of research on social cognition in people with temporal lobe epilepsy (TLE), the most prevalent type of epilepsy found in adults, with a focus on the components of this function that attract the most attention in the literature. In general, patients show deficits in various (affective and cognitive) Theory of Mind tasks. Emotion recognition deficits are common in patients with TLE and widespread across negative emotions, as well as deficits in cognitive empathy. Findings that overall suggest a certain degree of impairment of social cognition in patients with TLE can contribute to a deeper understanding of the socio-emotional difficulties they face in everyday life. In this sense, research on social cognition gives rise to the development and improvement of current epilepsy treatment practices, indicating the necessity of a stronger emphasis on psychosocial problems, in both diagnosis and treatment, in order to ensure holistic, comprehensive patient care

An assessment of emotion recognition in the prosodic characteristics of speech

Uvod: O emocionalnom stanju sagovornika može se zaključiti i na osnovu auditivne analize, odnosno, interpretacije specifičnih prozodijskih karakteristika govora. Iako manje istraženo u odnosu na sposobnost prepoznavanja emocija u izrazu lica, poteškoće u obradi govorne prozodije mogu predstavljati značajnu prepreku za adekvatno socijalno funkcionisanje i biti jezgro disfunkcije pojedinih poremećaja u razvojnom i odraslom dobu. Cilj: Konstrukcija i provera osnovnih psihometrijskih karakteristika testa za procenu uspešnosti prepoznavanja emocija na osnovu prozodijskih karakteristika govora. Metode: U konstrukciji seta sadržinski neutralnih rečenica, izgovorenih specifičnim tonom, a karakterističnim za svaku od šest osnovnih emocija uz kontrolni/neutralni stimulus, učestvovalo je šest glumaca. Nakon prve faze istraživanja u kojoj je učestvovalo pet ispitanika, odabrano je 35 stimulusa sa najvećim procentom tačnih odgovora, nakon čega je druga faza istraživanja sprovedena na uzorku od 45 ispitanika – 34 sa našeg govornog područja i 11 kojima srpski jezik nije maternji. Na osnovu procenta tačnosti prepoznavanja, za finalni set su odabrana 3 stimulusa po emociji, ukupno 21. Rezultati: Pouzdanost finalnog seta je α = 0,68. Empirijska distribucija mera značajno odstupa od modela normalne raspodele (W = 0,94, p = 0,02). Najveći procenat tačnosti prepoznavanja je zabeležen za emociju ljutnje (91,1%), dok se gađenje pokazalo kao najteže za prepoznati (53,3%). Nisu pronađene značajne razlike između ispitanika čiji se maternji jezici razlikuju (U = 135,00, p = 0,17). Postoji statistički značajna negativna povezanost između godina i ukupnog skora na testu (rs = -0,31, p = 0,04), dok se statistički značajne razlike između muškaraca i žena nisu ispoljile (p = 0,49). Zaključak: Test procene govorne prozodije, kao mera sposobnosti prepoznavanja emocionalnog stanja drugog, pokazuje potencijal za dalje korišćenje i razvoj.Introduction: The emotional state of the interlocutor can be deduced by way of auditory analysis, that is, the interpretation of specific prosodic characteristics of speech. Despite being less researched than the ability to recognize emotions in facial expressions, difficulties in processing speech prosody can be a major obstacle to adequate social functioning and the core of the dysfunction of certain disorders both during development and in adulthood. Aim: To develop and examine basic psychometric characteristics of the test for evaluating the success of emotion recognition based on the prosodic characteristics of speech. Methods: Six actors participated in the construction of a set of content- neutral sentences, spoken in a specific tone and representative of each of the six basic emotions with a control/neutral stimulus. Following the first phase of the research, in which five subjects took part, 35 stimuli with the highest percentage of correct answers were selected, after which the second phase of the research was conducted on a sample of 45 participants – 34 native speakers and 11 non-native speakers of Serbian. Based on the percentage of recognition accuracy, 3 stimuli per emotion, or 21 in total, were selected for the final set. Results: The reliability of the final set is α=.68. The empirical distribution of measures deviates significantly from the normal distribution model (W=0.94, p=.02). The highest percentage of recognition accuracy was recorded for anger (91.1%), while disgust showed to be the most difficult to recognize (53.3%). No significant differences were found among the participants who spoke different mother tongues (U=135.00, p=.17). There is a significant negative correlation between age and the total test score (rs= -.31, p=.04), while no significant differences between males and females were noticed (p=.49). Conclusion: As a measure of the ability to recognize the emotional state of another person, the speech prosody assessment test shows potential for further use and development

Enterocytes in Food Hypersensitivity Reactions

Food hypersensitivity reactions are adverse reactions to harmless dietary substances, whose causes are hidden within derangements of the complex immune machinery of humans and mammals. Until recently, enterocytes were considered as solely absorptive cells providing a physical barrier for unwanted lumen constituents. This review focuses on the enterocytes, which are the hub for innate and adaptive immune reactions. Furthermore, the ambiguous nature of enterocytes is also reflected in the fact that enterocytes can be considered as antigen-presenting cells since they constitutively express major histocompatibility complex (MHC) class II molecules. Taken together, it becomes clear that enterocytes have an immense role in maintaining oral tolerance to foreign antigens. In general, the immune system and its mechanisms underlying food hypersensitivity are still unknown and the involvement of components belonging to other anatomical systems, such as enterocytes, in these mechanisms make their elucidation even more difficult. The findings from studies with animal models provide us with valuable information about allergic mechanisms in the animal world, while on the other hand, these models are used to extrapolate results to the pathological conditions occurring in humans. There is a constant need for studies that deal with this topic and can overcome the glitches related to ethics in working with animals