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Comparative digestion of thermally treated vertebrates and invertebrates allergen pairs in real food matrix
No full textThe digestion stability of allergen pairs, tropomyosin, TM (fish and seafood allergen), and myosin light chain, MLC (chicken meat allergen) is compared among vertebrates and invertebrates in raw and cooked food matrix under standardized simulated in vitro gastrointestinal (GI) digestion. SDS-PAGE followed by multiple TM and MLC-specific antibodies in semidry WB revealed pepsin resistance of invertebrate TMs (abalone, oyster, shrimp) under diet-relevant conditions (raw, cooked). Vertebrate TMs (chicken, pork, beef) were less stable to digestion except that the raw chicken TM was observed pepsin resistant (not diet-relevant). Vertebrate (chicken) MLC was thermally stable. A new 28 kDa protein bound to anti-MLC antibody in cooked chicken and pork; could be the aggregates of MLC. Raw shrimp MLC showed pepsin resistance among invertebrates. A good correlation was observed between combined resistance of TM and MLC to GI digestion following the diet-relevant thermal treatment and reported protein allergenicity among vertebrates and invertebrates.Peer-reviewed version: [http://intor.torlakinstitut.com/handle/123456789/767
Determination of muscle fiber types expressing ANKRD2
Get PDFIntroduction: Ankyrin Repeat Domain 2 (ANKRD2) is expressed in skeletal muscle, where plays a role inmuscle development, differentiation and adaptation to stress. Human skeletal muscle consists of threemajor fiber types: type 1 (slow-twitch, oxidative), type 2A (fast-twitch, oxidative) and type 2X (fast-twitch,glycolytic). ANKRD2 is reported to be primarily expressed in type 1 myofibers. However, recent findingson human single myofibers and our study of chicken muscles have shown that this protein may also beexpressed in type 2A fibers. Hence, our objective was to examine whether ANKRD2 is present in humanfast, type 2A muscle fibers using immunohistochemistry.Methods: Samples of large leg musclessoleus, gastrocnemius, vastusintermedius and vastuslateralis wereobtained from human cadaveric tissue. Serial cryosections were independently stained with anti-ANKRD2and antibodies for different myosin heavy chain isoforms (6H1 for type 2X, BF35 for type 1 and 2A, antiMHCs for type 1 and anti-MHCf for type 2A and 2X fibers). Immunostained tissues were analyzed by fluorescent microscopy.Results: In addition to slow, type 1, ANKRD2 wasfound expressed in fast, type 2A myofibers, which bothhave oxidative metabolism. Further, we did not observe ANDRD2 expression in glycolytic, type 2Xmyiofibers. This pattern of ANKRD2 expression was consistent across all examined muscles.Conclusion: Our resultsimplicate that the regulatory mechanism of ANKRD2 expression in human skeletal muscle is associated with oxidative metabolism, rather than muscle contraction speed
Dynamic light scattering analysis of immune complexes in sera of rheumatoid arthritis patients
No full textThe size of circulating immune complexes (CICs) in rheumatoid arthritis (RA) could be an emerging criterion in disease diagnosis. This study analyzed size and electrokinetic potential of CICs from RA patients, healthy young adults, and RA patients age-matched controls aiming to establish their unique CIC features. Pooled CIC of 30 RA patients, 30 young adults, and 30 RA group's age-matched controls (middle-aged and oldеr healthy adults), and in vitro IgG aggregates from pooled sera of 300 healthy volunteers were tested using dynamic light scattering (DLS). Size distribution of CIC in healthy young adults exhibited high polydispersity. RA CIC patients and their age-matched control showed distinctly narrower size distributions compared with young adults. In these groups, particles clustered around two well-defined peaks. Particles of peak 1 were 36.1 ± 6.8 nm in RA age-matched control, and 30.8 ± 4.2 nm in RA patients. Particles of peak 2 of the RA age-matched control's CIC was 251.7 ± 41.2 nm, while RA CIC contained larger particles (359.9 ± 50.5 nm). The lower zeta potential of RA CIC, compared to control, indicated a disease-related decrease in colloidal stability. DLS identified RA-specific, but also age-specific distribution of CIC size and opened possibility of becoming a method for CIC size analysis in IC-mediated diseases
Erythrocyte fatty acid aberrations in Amyotrophic Lateral Sclerosis: Correlation with disease duration
Get PDFBackground: Recent literature data highlights metabolic changes in amyotrophic lateral sclerosis (ALS). To explore possible early metabolic changes, we aimed to analyse the fatty acids (FA) composition of erythrocytes in newly diagnosed als patients and to see whether fatty acid levels correlate with the ALSFRS-R score or disease duration. Methods: The severity of motor function involvement was assessed by the ALSFRS-R scale at the initial evaluation. The fatty acid profile of erythrocyte membranes was analysed by gas-liquid chromatography. The study comprised 26 clinically diagnosed als patients, with mean ALSFRS-R 38±8. The control group included 26 healthy volunteers. Results: Significantly higher levels of palmitic acid and total saturated FAs were detected in als patients. In als patients, total monounsaturated FA, palmitoleic, vaccenic, and oleic acid were also significantly increased. The levels of eicosapentaenoic acid, docosapentaenoic acid, total polyunsaturated FA (PUFA) and n-6 PUFA were significantly lower in als patients. Additionally, a-linolenic acid, the precursor of the n-3 PUFA family, was not detected in als patients. We found no significant correlation between the ALSFRS-R score and the abundance of individual FAs analysed. A moderate negative correlation was found between disease duration and DHA level, and a positive correlation was detected with MUFA. Conclusion: Experimental evidence presented may contribute to shaping a beneficial nutritional intervention
Antimikrobna antitela u fiziološkim i patološkim stanjima
No full textU ovom izlaganju biće predstavljeni naši dosadašnji rezultati na temu antimikrobnih antitela kod profesionalnih sportista, odraslih osoba koje se ne bave profesionalno sportom; promene nivoa antibakterijskih antitela pri starenju; poređenje specifičnosti serumskih i salivarnih IgA antitela kod mladih odraslih osoba, kao i nivoi ukupnih i antibakterijskih antitela u Covid-19 virusnoj infekciji i u sepsi. Nivoi i titri antimikrobnih antitela različitih klasa i potklasa određivani su ELISA testom iz uzoraka seruma, plazme ili salive. Ispitivano je vezivanje za izolovane antigene mikroorganizama (lipopolisaharid, peptidoglikan i zimozan) i za cele mikroorganizme, različitih vrsta i sojeva. Vezivanje prečišćenog salivarnog IgA za mikroorganizme praćeno je i protočnom citometrijom. Od rezultata izdvajamo da izlaganje fizičkom naporu visokog intenziteta dovodi do promena na nivou antibakterijskih antitela, i to u pravcu sveobuhvatnijeg prepoznavanja lipopolisaharida1. Suplementacija profesionalnih sportista određenim probioticima dovodi do održanja nivoa ukupnih salivarnih i serumskih IgA antitela, kao i IgG antitela specifičnih prema Enterococcus faecalis, koja su u ovoj populaciji pokazala sezonsku varijaciju. Pri starenju dolazi do smanjenja nivoa antipneumokoknih antitela i to zavisno od pola, pa su stariji mukarci naročito pogođeni ovim smanjenjem3. Salivarna IgA antitela razlikuju se po specifičnosti od serumskih IgA antitela i pružaju nespecifičnu zaštitu. Poređenje pacijenata sa sepsom, hospitalizovanih Covid-19 pacijenta i starosnih kontrola pokazalo je brojne razlike u nivoima kako ukupnih, tako i antibakterijskih antitela. Nivoi antibakterijskih antitela kod pacijentata koji nisu preživeli bili su niži nego kod onih koji su preživeli, što potvrđuje značaj ovih anititela za preživljavanje sepse
Sentinel Surveillance of Severe Acute Respiratory Infections in Serbia in 2022-2023 Season
No full textIntroduction:
Sentinel surveillance of severe acute respiratory infections
(SARI) was implemented in Serbia in November 2009. Nine
sentinel hospitals in 3 cities participated in the integrated SARI
surveillance system. Four of them are children’s hospitals. The
aim of this study is to provide a review of integrated sentinel
surveillance of SARI in Serbia in 2022-2023 influenza season,
from October 2022 to April 2023.
Methods:
Integrated Sentinel Surveillance of SARI has been done in
accordance with professional guidance for epidemiological
surveillance of influenza. Both epidemiological and virological
data were collected on a weekly basis. All specimens were tested
for influenza, SARS-CoV-2 and Respiratory syncytial virus
(RSV). Intensive care units (ICUs), pediatric and respiratory
disease wards were all represented. For laboratory confirmation,
Real time polymerase chain reaction (RT-PCR) was used.
Results:
Start of influenza season was registered in week 44/2022. A
total of 3027 SARI cases were reported with 100% recorded
age. Of these, 14% were 0-4 years old, 9% were 5-14, 7% were
15-29, 38% were 30-64 and 32% were 65. Among these cases,
607(20%) respiratory specimens were collected during the
surveillance period. The number of positive samples for
influenza was 208(34%), for SARS-CoV-2 was 37(6%) and
for RSV 12(2%). The highest proportion of laboratoryconfirmed
influenza cases was 58% in week 51/2022. then,
above 50% in week 7/2023 and 8/2023. All three influenza
viruses were confirmed: A(H1)pdm09, A(H3) and type B. B
viruses predominated, accounting for 38% of all sentinel SARI detections. There no were death associated to influenza, SARSCoV-
2 and RSV.
Conclusions:
Integration epidemiological with laboratory surveillance as
well as integrated sentinel SARI surveillance for influenza,
SARS -CoV-2 and RSV highlights the importance of maintaining
and improving national influenza surveillance capacity
especially in the frame of International Health Regulations.
Key messages:
Quality, representativeness and sustainability are the
guiding principles for integrated, SARS-CoV-2, RSV and
influenza sentinel SARI surveillance.
The threat of influenza remains, and it is essential for
countries to be vigilant for the emergence of non seasonal
influenza viruses of pandemic potential and prepare for the
next influenza season
A novel YtnP lactonase reduces the expression of p. aeruginosa MMA83 quorum sensing andvirulence factors gene expression
Get PDFIntroduction: Quorum quenching (QQ) isthe enzymatic degradation of cell-to-cellsignaling molecules.
In this study, the potential of the novel YtnP lactonase, the quorum quenching enzyme derived from S.
maltophilia, to reduce P. aeruginosa quorum sensing and virulence factor gene expression was investigated.
Methods: MMA83 culture (adjusted to 1.5x105 CFU/ml) was treated with recombinant YtnP lactonase
(final concentration 50 μg/ml) at 37°C for 12 hours under aeration. RNA isolation of the treated and untreated MMA83 culture was performed using the RNeasy Mini Kit (Qiagen, Germany) according to the
protocol. Quantitative reverse transcription-polymerase chain reaction (RT-qPCR), was used to analyze
the effect ofYtnP lactonase on the relative mRNA levels of the LasI/LasR, RhiI/RhiR, and PQS signaling network genes of P. aeruginosa MMA83 and virulence factor genes. The rpsL was used as an endogenous
control to normalize obtained data following the 2-ΔΔCt method.
Results: The QS genes belonging to three QS networks – LasI/LasR, RhiI/RhiR, and PQS of P. aeruginosa
MMA83 treated with YtnP lactonase were significantly downregulated. The RT -qPCR results show that
treatment with YtnP-lactonase decreased the relative mRNA levels of genes involved in the production
of elastase (lasB approximately 2-fold), alginate (algK approximately 2.2-fold), pyocyanin (phzM approximately 3.5-fold), pyoverdin (pvdS approximately 2-fold), and rhamnolipid (rhlC approximately 4-fold).
These results suggest that YtnP lactonase exerts an antivirulence effect at the transcription level.
Conclusion: YtnP lactonase, a quorum quenching (QQ) enzyme, has the potential to be used as an innovative enzyme-based antivirulence therapeutic to combat infections caused by P. aeruginosa
Epidemiological Predictors of Positive SARS-CoV-2 Polymerase Chain Reaction Test in Three Cohorts: Hospitalized Patients, Healthcare Workers, and Military Population, Serbia, 2020
Get PDFBackground: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its resulting coronavirus disease 2019 (COVID-19) has caused a fast-moving pandemic. Diagnostic testing, aimed to identify patients infected with SARS-CoV-2, plays a key role in controlling the COVID-19 pandemic in different populations. (2) Methods: This retrospective cohort study aimed to investigate predictors associated with positive polymerase chain reaction (PCR) SARS-CoV-2 test results in hospitalized patients, healthcare workers (HCWs), and military personnel (MP) during 2020, before the widespread availability of COVID-19 vaccines. Persons with a positive test result were compared with persons with a negative test result in three cohorts during the study period. (3) Results: A total of 6912 respondents were tested, and 1334 (19.3%) of them had positive PCR SARS-CoV-2 test results. Contact with a known COVID-19 case within 14 days (p < 0.001; OR: 1.48; 95% CI: 1.25–1.76), fever (p < 0.001; OR: 3.66; 95% CI: 3.04–4.41), cough (p < 0.001; OR: 1.91; 95% CI: 1.59–2.30), headache (p = 0.028; OR: 1.24; 95% CI: 1.02–1.50), and myalgia/arthralgia (p < 0.001; OR: 1.99; 95% CI: 1.65–2.42) were independently associated with positive PCR SARS-CoV-2 test results in the cohort of MP. Furthermore, fever (p < 0.001; OR: 2.75; 95% CI: 1.83–4.13), cough (p < 0.001; OR: 2.04; 95% CI: 1.32–3.13), headache (p = 0.008; OR: 1.76; 95% CI: 1.15–2.68), and myalgia/arthralgia (p = 0.039; OR: 1.58; 95% CI: 1.02–2.45) were independently associated with positive PCR SARS-CoV-2 test results in the cohort of HCWs. Moreover, independent predictors of positive PCR SARS-CoV-2 test results in hospitalized patients were contact with a known COVID-19 case within 14 days (p < 0.001; OR: 2.56; 95% CI: 1.71–3.83), fever (p < 0.001; OR: 1.89; 95% CI: 1.38–2.59), pneumonia (p = 0.041; OR: 1.45; 95% CI: 1.01–2.09), and neurological diseases (p = 0.009; OR: 0.375; 95% CI: 0.18–0.78). (4) Conclusions: According to data gathered from cohorts of hospitalized patients, HCWs, and MP, before the widespread availability of COVID-19 vaccines in Serbia, we can conclude that predictors of positive PCR SARS-CoV-2 test results in MP and HCWs were similar. Accurate estimates of COVID-19 in different population groups are important for health authorities
Food allergies on the rise: the role of anthropogenic chemicals
Get PDFFood allergies have increased dramatically in the last decade, especially in developedcountries. Food tolerance requires strict maintenance of a specific microbial portfolio inthe gastrointestinal tract, as changes in the gut microbiome can lead to its disruption,which in turn causes inflammation and pathogenic gut conditions leading to thedevelopment of food allergies. Any environmental factors that lead to a disturbanceand/or malfunction of the gastrointestinal tract and digestive performance favor thedevelopment of food allergies.Based on that, what do we know about the role of increasing anthropogenic chemicals,including emerging ones, resulting from the new global situation?There is awareness that their effects are multifaceted, e.g., chemicals affect the growth ofplants and animals and thus the quality of the food produced. In addition, chemicals affectour food during its production and processing, but also affect our body andgastrointestinal tract. It is time to fill the knowledge gaps and understand how theseinteractions between environmental triggers such as industrial and traffic pollution,transition and heavy metals, pesticides, chemtrails, etc., affect food allergens and theirallergenicity, adjuvant effects, and the increasing prevalence of food allergies.Some improvements in this area are already being made through advances in ‘omics’technologies (i.e., proteomics, genomics, metabolomics) and systems biology approachesthat will hopefully provide a scientific understanding of the relationship betweenincreasing food allergies and the increasingly present wide variety of anthropogenicchemicals in our environment
Thymic changes as a contributing factor in the increased susceptibility of old Albino Oxford rats to EAE development
Get PDFThe study was aimed to examine putative contribution of thymic involution to ageing-associated increase in susceptibility of Albino Oxford (AO) rats to the development of clinical EAE, and vice versa influence of the disease on the progression of thymic involution. To this end we examined (i) the parameters of thymocyte negative selection efficacy, the thymic generation of CD4+CD25+Foxp3+ T regulatory cells (Tregs) and thymic capacity to instruct/predetermine IL-17-producing T-cell differentiation, and thymopietic efficacy-associated accumulation of “inflammescent” cytotoxic CD28- T cells in the periphery, and (ii) the key underlying mechanisms in young and old non-immunised AO rats and their counterparts immunised for EAE (on the 16th day post-immunisation when the disease in old rats reached the plateau) using flow cytometry analysis and/or RT-qPCR. It was found that thymic involution impairs: (i) the efficacy of negative selection (by affecting thymocyte expression of CD90, negative regulator of selection threshold and the expression of thymic stromal cell integrity factors) and (ii) Treg generation (by diminishing expression of cytokines supporting their differentiation/maturation). Additionally, the results suggest that thymic involution facilitates CD8+ T-cell differentiation into IL-17-producing cells (previously linked to the development of clinical EAE in old AO rats). Furthermore, they confirmed that ageing-related decrease in thymic T-cell output (as indicated by diminished frequency of recent thymic emigrants in peripheral blood) resulted in the accumulation of CD28- T cells in peripheral blood and, upon immunisation, in the target organ. On the other hand, the development of EAE (most likely by increasing circulatory levels of proinflammatory cytokines) contributed to the decline in thymic output of T cells, including Tregs, and thereby to the progression/maintenance of clinical EAE. Thus, in AO rats thymic involution via multi-layered mechanisms may favour the development of clinically manifested autoimmunity, which, in turn, precipitates the thymus atrophy