Imunoreaktivnost na proteine hrane u etiopatogenezi rekurentnih oralnih ulceracija

Based on a complete analysis of the results , we came to the following conclusions: 1. Characteristics of ROU: • The experimental group consisted of 38% men and 62% women. • The average age of the experimental group was 40.32 years. • The most common localization (53%) of ROU is the buccal and labial mucosa , then the tong (14%) and the whole mouth in 13% of patients. • In 86% of cases, the characteristics of ROU corresponded to clinical picture of aphthae minor, while in 14% of cases corresponded to aphthae major. • Over half of the patients (58%) at the time of the physical examination had 1-5 ulcerations in the oral cavity, 32% had from 5-10, while 10% had more than 10 ulcerations in the oral cavity. • In 52% of patients, ROU first emerged several years ago, in 40% of them for more than a year, while in 6% change first occurred in the period of 1-6 months ago, only one respondent (2%) stated that the ulcers first appeared over a period of 6-12 months before. • ROU relapse appeared from one month in 40% of patients, 30% of them stated incidence of recurrence of 2-5 times in 6 months, while in 18% of patients these changes occurred several times per month, the lowest number of patients (12%) stated that the recurrence comes from 1-3 times per year. • In the 86% of patients ROU heal over a period of 7-14 days , while in 14% of cases, heal more than one month . 2. Sensitization to food proteins: • In 14 % of patients with ROU elevated humoral immunoreactivity to gliadin is registered. • Elevated humoral immunoreactivity to the total cow milk protein was detected in more than half (72 %) patients with ROU. • Serum antibodies in the elevated concentration was present against: -α - casein in 72%, -β- casein in 62%, -κ- casein in 52%, -α - lactalbumin in 18%, - lactoferrin in 20%, -β - lactoglobulin A and 24% in -β - lactoglobulin B in 30% of patients. 3. Based on these results we conclude that cow's milk protein may be one of the etiological factors in the development of ROU .Na osnovu kompletne analize dobijenih rezultata, došli smo do sledećih zaključaka: 1. Karakteristike ROU: • Eksperimentalnu grupu je činilo 38% muškaraca i 62% žena. • Prosečna starost eksperimentalne grupe je iznosila 40.32 godine. • Najzastupljenija lokalizacija (53%) ROU su bukalna i labijalna sluzokoža, zatim jezik sa 14% i cela usta u 13% pacijenata. • U 86% slučajeva karakteristike ROU su odgovarale kliničkoj slici afta minor, dok su u 14% slučajeva odgovarale aftama major. • Više od polovine pacijenata (58%) je u momentu kliničkog pregleda imalo od 1-5 ulceracija u usnoj duplji, 32% je imalo od 5-10 promena, dok je 10% imalo više od 10 ulceracija u ustima. • Kod 52% pacijenata, ROU su se prvi put pojavile pre više godina, kod 40% njih pre više od godinu dana, dok su se u 6% slučajeva promene prvi put javile u periodu od pre 1-6 meseci; samo jedan ispitanik (2%) je izjavio da su se ulceracije prvi put pojavile u periodu od pre 6-12 meseci. • Recidiv ROU se javljao jednom mesečno u 40% pacijenata, 30% njih je navelo da je učestalost recidiva od 2-5 puta u toku 6 meseci, dok su se u 18% njih promene javljale više puta mesečno; najmanji broj pacijenata (12%) je izjavio da do recidiva dolazi od 1-3 puta u toku jedne godine. • U 86% pacijenata ROU zarastaju u periodu od 7-14 dana, dok u 14% slučajeva zarastaju duže od mesec dana. 2. Preosetljivost na proteine hrane: • Kod 14% ispitanih pacijenata sa ROU registrovana je povišena humoralna imunoreaktivnost na gliadin. • Povišena humoralna imunoreaktivnost na ukupne proteine kravljeg mleka je detektovana kod više od polovine (72%) pacijenata sa ROU. • Serumska antitela su u povišenoj koncentraciji bila prisutna na: - α-kazein u 72%, - β-kazein u 62%, - κ-kazein u 52%, - α-laktalbumin u 18%, - laktoferin u 20%, - β-laktoglobulin A u 24% i - β-laktoglobulin B u 30%. 3. Na osnovu dobijenih rezultata konstatujemo da proteini kravljeg mleka mogu biti jedan od etioloških faktora u nastanku ROU

The absence of immunoreactivity to donkey’s milk in patients with recurrent aphthous ulcers and immunoreactivity to cow’s milk

Despite the numerous benefits of milk constituents for human health a considerable number of the general population follow a milk-restricted diet due to clinically confirmed or self-assessed adverse reactions to cow’s milk consumption. Recurrent aphthous ulcers (RAU) are currently one of the most common oral disorders, with a worldwide distribution and insufficiently defined etiology, which, among other factors, implies the immunological reaction to food proteins. The aim of this study was to determine the immune-reactivity to donkey’s milk proteins in patients with RAU and compare it to the reactivity towards the proteins from cow’s and goat’s milks, in a set of simultaneous experiments. Levels of serum IgA, IgG and IgE antibodies to the same quantity of the examined antigens were determined by enzyme-linked immunosorbent assay. The results indicate that patients with RAU with increased immunity to cow’s milk proteins could consider the use of donkey’s milk as the best protein source

Celiac disease-specific and inflammatory bowel disease-related antibodies in patients with recurrent aphthous stomatitis

The etiology of recurrent aphthous stomatitis (RAS) remains unknown. RAS can be presented as primary, idiopathic condition and as a secondary RAS, which is associated with a systemic disease. The aim of our study was to evaluate the presence and concentrations of antibodies specific for celiac disease (CeD) and antibodies related to inflammatory bowel diseases (IBD) in patients with RAS without gastrointestinal symptoms. Antibodies against tissue transglutaminase (anti-tTG), deaminated gliadin peptides (DGP), deaminated gliadinanalogous fragments (anti-GAF-3X) and Saccharomyces cerevisiae (ASCA) were determined by ELISA and anti-neutrophil cytoplasmic antibodies (ANCA) by indirect immunoflurescence (IIF) in 57 patients with RAS and 60 control subjects. The prevalence of CeD specific antibodies did not differ between RAS patients and controls. However, the concentrations of IgA anti-tTG, IgA anti-GAF-3X antibodies in patients with RAS were significantly higher compared to controls (p = 0.002 and p = 0.04 respectively). Histological changes consistent with CeD were confirmed by duodenal biopsy in one RAS patient with highly positive IgA anti-tTG, anti-GAF-3X and anti-DGP antibodies. Higher prevalence along with higher concentrations of IgG ASCA were found in RAS patients compared to controls (p lt 0.01). Patients with positive IgG ASCA in the absence of clinical symptoms decided not to pursue any further testing. Dysfunction of oral mucosa and the exposure to various antigens might be a reason for the loss of tolerance resulting in increased production of autoantibodies. It seems likely that antibodies are markers of aberrant immune response, rather than key effectors involved in the pathogenesis of the disease

Lactobacilli hydrolysis of cows' milk proteins abrogates their humoral immunoreactivity in patients with immune-mediated diseases

The level of humoral immunoreactivity to total cows' milk proteins (TCMP) in sera of patients suffering from recurrent oral ulcerations, gastrointestinal diseases or haematological malignancies was investigated. TCMP were also hydrolysed with two different species of lactobacilli and dramatic changes in the levels of specific IgG and IgE were found with statistically significant decreases in the levels of specific antibodies in sera from all patient groups. The levels below cut-off values of IgG specific for TCMP hydrolysates were detected in sera from all patients, while values of IgE for hydrolysates obtained with Lactobacillus helveticus BGRA43 and Lactobacillus zeae LMG17315were below cut-off in 85% and 97% of patients, respectively. Competitive ELISA confirmed the specificity of antibodies for immunogenic TCMP epitopes, demonstrating that lactobacilli hydrolyse TCMP by degrading immunogenic epitopes, and could therefore be used in processing of milk proteins to obtain products suitable for patients with altered immune response on TCMP.This is the peer reviewed version of the paper: Vukotic, G., Matic, I., Begovic, J., Besu, I., Kojic, M., Djokic, J., Juranic, Z., & Strahinic, I. (2016). Lactobacilli hydrolysis of cows’ milk proteins abrogates their humoral immunoreactivity in patients with immune-mediated diseases. International Dairy Journal, 63, 1–7. [https://doi.org/10.1016/j.idairyj.2016.07.009

Different levels of humoral immunoreactivity to different wheat cultivars gliadin are present in patients with celiac disease and in patients with multiple myeloma

<p>Abstract</p> <p>Background</p> <p>Immunity to food antigens (gliadin, cow's milk proteins) is in the centre of the attention of modern medicine focused on the prevention of diseases, prevention which is based on the use of appropriate restriction diet. Detection of the enhanced levels of the immune reactions to antigen(s) present in food is from this point of view of great importance because there are reports that some of health disturbances, like celiac disease (CD) and some premalignant conditions, like monoclonal gammopathy of undetermined significance (MGUS), were vanished after the appropriate restriction diets.</p> <p>It is well known that gliadin is toxic to small bowel mucosa of relatively small population of genetically predisposed individuals, who under this toxic action develop celiac disease (CD). As the quantity of immunogenic gliadin could vary between different wheat species, the first aim of this work was to determine the percentage of immunogenic gliadin in ten bread wheat cultivars and in three commercially grown durum wheat cultivars. The second part of the study was initiated by results of previous publication, reporting that sera of some of multiple myeloma (MM) patients showed the presence of elevated levels of anti-gliadin IgA, without the enhanced levels of anti-gliadin IgG antibodies, determined with commercial ELISA test. It was designed to assess is it possible to reveal is there any hidden, especially anti-gliadin IgG immunoreactivity, in serum of mentioned group of patients. For this purpose we tested MM patients sera, as well as celiac disease (CD) patients sera for the immunoreaction with the native gliadin isolated from wheat species used for bread and pasta making in corresponding geographic region.</p> <p>Results</p> <p>Gliadin was isolated from wheat flour by two step 60% ehanolic extraction. Its content was determined by commercial R5 Mendez Elisa using PWG gliadin as the standard. Results obtained showed that immunogenic gliadin content varies between 50.4 and 65.4 mg/g in bread wheat cultivars and between 20 and 25.6 mg/g in durum wheat cultivars.</p> <p>Anti-gliadin IgA and IgG immunoreactivity of patients' sera in (IU/ml) was firstly determined by commercial diagnostic Binding Site ELISA test, and then additionally by non-commercial ELISA tests, using standardized ethanol wheat extracts -gliadin as the antigen.</p> <p>In both patients groups IgA immunoreactivity to gliadin from different cultivars was almost homogenous and in correlation with results from commercial test (except for one patient with IgA(λ) myeloma, they were more then five times higher). But, results for IgG immunoreactivity were more frequently inhomogeneous, and especially for few MM patients, they were more then five times higher and did not correlate with results obtained using Binding Site test.</p> <p>Conclusion</p> <p>Results obtained showed different content of immunogenic gliadin epitopes in various species of wheat.</p> <p>They also point for new effort to elucidate is there a need to develop new standard antigen, the representative mixture of gliadin isolated from local wheat species used for bread production in corresponding geographic region for ELISA diagnostic tests.</p

Self-Heating Flower-like Nanoconstructs with Limited Incorporation of Yttrium in Maghemite: Effect of Chemical Composition on Heating Efficiency, Cytotoxicity and Genotoxicity

Partial cation substitution can significantly change the physical properties of parent compounds. By controlling the chemical composition and knowing the mutual relationship between composition and physical properties, it is possible to tailor the properties of materials to those that are superior for desired technological application. Using the polyol synthesis procedure, a series of yttrium-substituted iron oxide nanoconstructs, γ-Fe2−xYxO3 (YIONs), was prepared. It was found that Y3+ could substitute Fe3+ in the crystal structures of maghemite (γ-Fe2O3) up to a limited concentration of ~1.5% (γ-Fe1.969Y0.031O3). Analysis of TEM micrographs showed that crystallites or particles were aggregated in flower-like structures with diameters from 53.7 ± 6.2 nm to 97.3 ± 37.0 nm, depending on yttrium concentration. To be investigated for potential applications as magnetic hyperthermia agents, YIONs were tested twice: their heating efficiency was tested and their toxicity was investigated. The Specific Absorption Rate (SAR) values were in the range of 32.6 W/g to 513 W/g and significantly decreased with increased yttrium concentration in the samples. Intrinsic loss power (ILP) for γ-Fe2O3 and γ-Fe1.995Y0.005O3 were ~8–9 nH·m2/Kg, which pointed to their excellent heating efficiency. IC50 values of investigated samples against cancer (HeLa) and normal (MRC-5) cells decreased with increased yttrium concentration and were higher than ~300 μg/mL. The samples of γ-Fe2−xYxO3 did not show a genotoxic effect. The results of toxicity studies show that YIONs are suitable for further in vitro/in vivo studies toward to their potential medical applications, while results of heat generation point to their potential use in magnetic hyperthermia cancer treatment or use as self-heating systems for other technological applications such as catalysis

Detection of humoral and cellular immune response to anti-SARS-CoV-2 BNT162b2 vaccine in breastfeeding women and naïve and previously infected individuals

This study explored humoral and cellular responses to anti-SARS-CoV-2 BNT162b2 mRNA vaccine in breastfeeding women and naïve and seropositive individuals in the first six months after vaccination.Sixty-one volunteers vaccinated with two doses of the BNT162b2 mRNA vaccine were enrolled in the study. In-house developed ELISA was used for the quantification of SARS-CoV-2 RBD-specific antibodies. Cell surface marker expression and intracellular IFN-γ analysis were carried out by flow cytometry. The concentrations of IFN-γ, IL-6 and TNF were determined by ELISA. A significant rise in anti-RBD IgG antibody levels was observed 14 days after the first vaccine dose (p < 0.0001) in serum and milk. The expression of CD28 on CD4+ T cells was significantly higher compared to baseline (p < 0.05). There was a significant increase (p ≤ 0.05) in B cell lymphocyte subset after revaccination, and increased percentage of CD80+ B cells. The expression of IFN-γ in peripheral blood lymphocytes, CD3+ T cells and serum was significantly increased (p < 0.05). No significant difference in immune response was observed between breastfeeding women and other study participants. The anti-SARS-CoV-2 BNT162b2 mRNA vaccine-induced measurable and durable immune response in breastfeeding women and in naïve and previously infected individuals

Evaluation of cytokine expression and circulating immune cell subsets as potential parameters of acute radiation toxicity in prostate cancer patients

One of the challenges of radiation oncology in the era of personalized medicine is identification of biomarkers associated with individual radiosensitivity. The aim of research was to evaluate the possible clinical value of the associations between clinical, physical, and biological factors, and risk for development of acute radiotoxicity in patients with prostate cancer. The study involved forty four patients treated with three-dimensional conformal radiotherapy. The concentrations of IL-1β, IL-2, IL-6, IFN-γ and TGF-β1 were assessed before radiotherapy, after 5th, 15th and 25th radiotherapy fractions, at the end, and 1 month after the end of radiotherapy. Cytokine gene expression was determined in peripheral blood mononuclear cells. The univariate analysis of circulating cytokine levels during radiotherapy showed that increased serum concentrations of IL-6 were significantly associated with higher grade of acute genitourinary toxicity. The multivariate analysis demonstrated that increased level of IL-6 during the radiotherapy was significantly associated with higher grade of acute genitourinary toxicity across treatment. TGF-β expression levels significantly decreased during course of radiotherapy. Research indicates that changes in circulating cytokine levels might be important parameter of radiotoxicity in patients with prostate cancer. These findings suggest that future studies based on multi-parameter examination are necessary for prediction of individual radiosensitivity

Immunoreactivity to food protein in pathogenesis of recurrent oral ulceration

Nа оsnоvu kоmplеtnе аnаlizе dоbiјеnih rеzultаtа, dоšli smо dо slеdеćih zаklјučаkа: 1. Kаrаktеristikе RОU: • Еkspеrimеntаlnu grupu је činilо 38% muškаrаcа i 62% žеnа. • Prоsеčnа stаrоst еkspеrimеntаlnе grupе је iznоsilа 40.32 gоdinе. • Nајzаstuplјеniја lоkаlizаciја (53%) RОU su bukаlnа i lаbiјаlnа sluzоkоžа, zаtim јеzik sа 14% i cеlа ustа u 13% pаciјеnаtа. • U 86% slučајеvа kаrаktеristikе RОU su оdgоvаrаlе kliničkој slici аftа minоr, dоk su u 14% slučајеvа оdgоvаrаlе аftаmа mајоr. • Višе оd pоlоvinе pаciјеnаtа (58%) је u mоmеntu kliničkоg prеglеdа imаlо оd 1-5 ulcеrаciја u usnој duplјi, 32% је imаlо оd 5-10 prоmеnа, dоk је 10% imаlо višе оd 10 ulcеrаciја u ustimа. • Kоd 52% pаciјеnаtа, RОU su sе prvi put pојаvilе prе višе gоdinа, kоd 40% njih prе višе оd gоdinu dаnа, dоk su sе u 6% slučајеvа prоmеnе prvi put јаvilе u pеriоdu оd prе 1-6 mеsеci; sаmо јеdаn ispitаnik (2%) је izјаviо dа su sе ulcеrаciје prvi put pојаvilе u pеriоdu оd prе 6-12 mеsеci. • Rеcidiv RОU sе јаvlјао јеdnоm mеsеčnо u 40% pаciјеnаtа, 30% njih је nаvеlо dа је učеstаlоst rеcidivа оd 2-5 putа u tоku 6 mеsеci, dоk su sе u 18% njih prоmеnе јаvlјаlе višе putа mеsеčnо; nајmаnji brој pаciјеnаtа (12%) је izјаviо dа dо rеcidivа dоlаzi оd 1-3 putа u tоku јеdnе gоdinе. • U 86% pаciјеnаtа RОU zаrаstајu u pеriоdu оd 7-14 dаnа, dоk u 14% slučајеvа zаrаstајu dužе оd mеsеc dаnа. 2. Prеоsеtlјivоst nа prоtеinе hrаnе: • Kоd 14% ispitаnih pаciјеnаtа sа RОU rеgistrоvаnа је pоvišеnа humоrаlnа imunоrеаktivnоst nа gliаdin. • Pоvišеnа humоrаlnа imunоrеаktivnоst nа ukupnе prоtеinе krаvlјеg mlеkа је dеtеktоvаnа kоd višе оd pоlоvinе (72%) pаciјеnаtа sа RОU. • Sеrumskа аntitеlа su u pоvišеnој kоncеntrаciјi bilа prisutnа nа: - α-kаzеin u 72%, - β-kаzеin u 62%, - κ-kаzеin u 52%, - α-lаktаlbumin u 18%, - lаktоfеrin u 20%, - β-lаktоglоbulin А u 24% i - β-lаktоglоbulin B u 30%. 3. Nа оsnоvu dоbiјеnih rеzultаtа kоnstаtuјеmо dа prоtеini krаvlјеg mlеkа mоgu biti јеdаn оd еtiоlоških fаktоrа u nаstаnku RОU.Based on a complete analysis of the results , we came to the following conclusions: 1. Characteristics of ROU: • The experimental group consisted of 38% men and 62% women. • The average age of the experimental group was 40.32 years. • The most common localization (53%) of ROU is the buccal and labial mucosa , then the tong (14%) and the whole mouth in 13% of patients. • In 86% of cases, the characteristics of ROU corresponded to clinical picture of aphthae minor, while in 14% of cases corresponded to aphthae major. • Over half of the patients (58%) at the time of the physical examination had 1-5 ulcerations in the oral cavity, 32% had from 5-10, while 10% had more than 10 ulcerations in the oral cavity. • In 52% of patients, ROU first emerged several years ago, in 40% of them for more than a year, while in 6% change first occurred in the period of 1-6 months ago, only one respondent (2%) stated that the ulcers first appeared over a period of 6-12 months before. • ROU relapse appeared from one month in 40% of patients, 30% of them stated incidence of recurrence of 2-5 times in 6 months, while in 18% of patients these changes occurred several times per month, the lowest number of patients (12%) stated that the recurrence comes from 1-3 times per year. • In the 86% of patients ROU heal over a period of 7-14 days , while in 14% of cases, heal more than one month . 2. Sensitization to food proteins: • In 14 % of patients with ROU elevated humoral immunoreactivity to gliadin is registered. • Elevated humoral immunoreactivity to the total cow milk protein was detected in more than half (72 %) patients with ROU. • Serum antibodies in the elevated concentration was present against: -α - casein in 72%, -β- casein in 62%, -κ- casein in 52%, -α - lactalbumin in 18%, - lactoferrin in 20%, -β - lactoglobulin A and 24% in -β - lactoglobulin B in 30% of patients. 3. Based on these results we conclude that cow's milk protein may be one of the etiological factors in the development of ROU