Role of gut microbiota and gut-associated lymphoid tissue in resistance of Albino Oxford rats to experimental autoimmune encephalomyelitis induction

Multipla skleroza je hronična, inflamacijska, neurodegenerativna, demijelinizirajuća bolest centralnog nervnog sistema (CNS). Etiologija multiple skleroze još uvek nije poznata, ali je sve više podataka koji pokazuju ulogu crevne mikrobiote u patogenezi bolesti. Više podataka o značaju crevne mikrobiote u autoimunosti CNS dobijeno je istraživanjima na eksperimentalnom autoimunskom encefalomijelitisu (EAE), životinjskom modelu multiple skleroze. U EAE-u se autoimunski odgovor protiv CNS-a pokreće u limfnim čvorovima koji dreniraju mesto imunizacije, gde se CD4+ T ćelije specifične za antigene CNS-a aktiviraju i diferenciraju u Th (engl. T helper) 1 ćelije koje produkuju IFN-γ i Th17 ćelije koje produkuju IL-17. Kada dospeju u CNS ove Th ćelije bivaju reaktivirane od strane lokalnih antigen-prezentujućih ćelija. Nakon reaktivacije, encefalitogene Th ćelije pokreću i održavaju inflamaciju unutar CNS-a. Albino oksford (AO) pacovi su veoma otporni na indukciju EAE-a. S druge strane, Dark Agouti (DA) pacovi razvijaju EAE čak i nakon blage imunizacije. U dosadašnjim istraživanjima je pokazano da se AO pacovi u odnosu na DA pacove razlikuju po slaboj aktivaciji Th1 i Th17 ćelija u relevantnim limfnim čvorovima u odgovoru na encefalitogenu imunizaciju. Takođe, pokazano je da relativno mali broj encefalitogenih ćelija infiltrira CNS AO pacova. Do sada nije vršeno ispitivanje uloge GALT-a i mikrobiote creva u otpornosti pacova soja AO na indukciju EAE-a. Sve je više istraživanja koja pokazuju da mikroorganizmi creva imaju važnu ulogu u regulaciji imunskog odgovora kroz interakciju sa limfnim tkivom creva (GALT, engl. gut-associated lymphoid tissue). U istraživanjima na EAE-u je pokazano da određeni mikroorganizmi creva pospešuju tok bolesti, dok drugi ostvaruju terapeutske efekte modulacijom odnosa Th1 i Th17 encefalitogenih ćelija i regulatornih T ćelija (Treg). Poznato je i da encefalitogene CD4+ T ćelije migriraju u GALT, uključujući mezenterične limfne čvorove i Pejerove ploče gde se mogu diferencirati u Treg...Multiple sclerosis is a chronic inflammatory neurodegenerative demyelinating disease of the central nervous system (CNS). Etiology of multiple sclerosis is still unknown, but data are showing that gut microbiota plays an important role in the pathogenesis of this disease. Information about the significance of gut microbiota in CNS autoimmunity has been dominantly obtained in studies in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. The autoimmune response against CNS starts in lymph nodes draining the site of immunization in EAE. There, CD4+ T cells are being activated and differentiated into Th1 (helper T cells) and Th17 cells that produce IFN-γ and IL-17, respectively. When Th cells arrive in the CNS, they are reactivated by local antigen-presenting cells. After the reactivation, these encephalitogenic T cells initiate and propagate CNS inflammation. Albino Oxford (AO) rats are highly resistant to EAE induction. On the other hand, Dark Agouti (DA) rats develop EAE even after the mild immunization. In our earlier studies it was shown that AO rats had less activated Th1 and Th17 cells in the relevant lymph nodes in response to encephalitogenic immunization, unlike DA rats. Also, it was shown that relatively small number of encephalitogenic cells infiltrated CNS in AO rats. Up until now, the studies of role of GALT and gut microbiota in resistance of AO rats in EAE induction have not been conducted. It has been increasingly appreciated that gut microbiota plays an important role in the regulation of the immune response through interaction with the cells of gut-associated lymphoid tissue (GALT). In EAE studies specific gut microorganisms have been proposed to promote the disease, while others have been shown to have therapeutic effects by modulating ratio and activity of encefalitogenic Th1 and Th17 cells and regulatory T cells (Treg). Encefalitogenic cells can also migrate to GALT, including mesenteric lymph nodes and Peyer’s patches, where they can be re-differentiated into Treg..

Use of honeybees (Apis mellifera L.) as bioindicators of spatial variations and origin determination of metal pollution in Serbia

Honeybees have been proposed and used as bioindicators for the last few decades, because of their nature. Until now they have mostly been used to determine the present pollution and to distinguish the differences between the sampling locations and the sampling periods. With the use of multivariate statistical methods honeybees can also be used to distinguish the origin of this pollution. In this study the concentrations of Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, Mg, Mn, Na, Ni, Sr and Zn were measured in the bodies of adult honeybees collected from nine different apiaries in Serbia. With the help of the statistical methods it was established that the least polluted area was the one that has no industrial activities or the intense traffic nearby. The most polluted was the urban region, followed by a region close to thermal power plants and ash disposal site. Using PCA and CA the origin of the analyzed metals were proposed. It was suggested that Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, Mn and Ni have anthropogenic origin mainly from the intensive agriculture, traffic and coal combustion.Supplementary material: [http://cherry.chem.bg.ac.rs/handle/123456789/3312

Supplementary data for the article: Zarić, N. M.; Ilijević, K.; Stanisavljević, L.; Gržetić, I. Use of Honeybees (Apis Mellifera L.) as Bioindicators of Spatial Variations and Origin Determination of Metal Pollution in Serbia. Journal of the Serbian Chemical Society 2018, 83 (6), 773–784. https://doi.org/10.2298/JSC171110018Z

Supplementary material for: [https://doi.org/10.2298/JSC171110018Z]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2179

Supplementary material for the article: Zaric, N. M.; Deljanin, I.; Ilijević, K.; Stanisavljević, L.; Ristić, M.; Gržetić, I. Assessment of Spatial and Temporal Variations in Trace Element Concentrations Using Honeybees (Apis Mellifera) as Bioindicators. PeerJ 2018, 2018 (7). https://doi.org/10.7717/peerj.5197

Supplementary material for: [https://doi.org/10.7717/peerj.5197]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2183

Supplementary material for the article: Zaric, N. M.; Deljanin, I.; Ilijević, K.; Stanisavljević, L.; Ristić, M.; Gržetić, I. Assessment of Spatial and Temporal Variations in Trace Element Concentrations Using Honeybees (Apis Mellifera) as Bioindicators. PeerJ 2018, 2018 (7). https://doi.org/10.7717/peerj.5197

Supplementary material for: [https://doi.org/10.7717/peerj.5197]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/2183

A simple and efficient DNA isolation method for Ornithogalum L. species (Hyacinthaceae, Asparagales)

We report an efficient, simple and cost-effective protocol for the isolation of genomic DNA from Ornithogalum species. Our modification of the standard CTAB protocol includes two polyphenol adsorbents (insoluble PVPP and activated charcoal), high NaCl concentrations (4 M) for removing polysaccharides, and addition of phenol to remove proteins and other contaminants. DNA yield obtained with our protocol was 223 and 312 μg DNA g-1 of dry leaf tissue. The absorbance ratio 260/280 nm was 1.879 (O. refractum) and 1.753 (O. sibthorpii), and the absorbance ratio 260/230 nm was 1.779 (O. refractum) and 1.545 (O. sibthorpii), revealing lack of contamination. PCR amplifications of one nuclear marker (26S rDNA) indicated that this DNA isolation protocol may be used for Ornithogalum plants containing many interfering compounds for further analyses in population genetics and phylogeographic studies

Antioxidant activity, the content of total phenols and flavonoids in the ethanol extracts of Mentha longifolia (L.) Hudson dried by the use of different techniques

In this study, we have examined the yield of extracted substances obtained by means of extraction using 70 % ethanol (v/v), the content of total phenols and flavonoids, as well as the antioxidant activity of the extracts obtained from the samples of the herbs dried by means of different techniques. Wild mint Mentha longifolia (L.) Hudson was dried naturally in a laboratory oven at a temperature of 45 °C and in an absorptive low temperature condensation oven at 35°C. The highest yield of extracts was obtained from the naturally dried herbs and the lowest from the herbs dried in the low temperature condensation drying oven. The content of total phenols and flavonoids was determined by spectrophotometric methods with an FC reagent and by the complexation reaction with aluminium-chloride, respectively. The extract of the naturally dried herbs had the highest overall content of phenols (113.8±2.0 mg of gallic acid/g of the dry extract) and flavonoids (106.7±0.3 mg of rutin/g of the dry extract). The highest antioxidant activity determined by the FRAP and DPPH assay was determined in the extracts obtained from naturally dried herbs (2.76±0.15 mmol Fe2+/mg of the dry extract and EC50=0.022±0.001 mg/ml), while the lowest was obtained from the extracts of herbs dried in the laboratory oven (1.13±0.11 mmol Fe2+/mg of the dry extract and EC50=0.033±0.001 mg/ml). The HPLC-DAD analysis result show that the greatest content of phenolic compounds show extract obtained from naturally dried plant material. The dominant phenolic component in the all extracts is Kaempferol 3-O-glucoside. The content of all phenolic compound strongly depend on the drying conditions

Stabilization of Black Locust Flower Extract via Encapsulation Using Alginate and Alginate–Chitosan Microparticles

Black locust flower extract contains various polyphenols and their glucosides contribute to the potential health benefits. After intake of these bioactive compounds and passage through the gastrointestinal tract, their degradation can occur and lead to a loss of biological activity. To overcome this problem, the bioactive compounds should be protected from environmental conditions. This study aimed to encapsulate the black flower extract in the microparticles based on biodegradable polysaccharides, alginate, and chitosan. In the extract, the total antioxidant content was found to be 3.18 ± 0.01 g gallic acid equivalent per 100 g of dry weight. Also, the presence of lipids (16), phenolics (27), organic acids (4), L-aspartic acid derivative, questinol, gibberellic acid, sterol, and saponins (2) was confirmed using the UHPLC–ESI–MS analysis. In vitro assays showed that the extract has weak anti-α-glucosidase activity and moderate antioxidant and cytotoxic activity against the HeLa cell line. The extrusion method with secondary air flow enabled the preparation of microparticles (about 270 μm) encapsulated with extract. An encapsulation efficiency of over 92% was achieved in the alginate and alginate–chitosan microparticles. The swelling study confirmed a lower permeability of alginate–chitosan microparticles compared with alginate microparticles. For both types of microparticles, the release profile of antioxidants in the simulated gastrointestinal fluids at 37 °C followed the Korsmeyer–Peppas model. A lower diffusion coefficient than 0.5 indicated the simple Fick diffusion of antioxidants. The alginate–chitosan microparticles enabled a more sustained release of antioxidants from extract compared to the alginate microparticles. The obtained results indicated an improvement in the antioxidant activity of bioactive compounds from the extract and their protection from degradation in the simulated gastric conditions via encapsulation in the polymer matrixes. Alginate–chitosan showed slightly slower cumulative antioxidant release from microparticles and better antioxidant activity of the extract compared to the alginate system. According to these results, alginate–chitosan microparticles are more suitable for further application in the encapsulation of black locust flower extract. Also, the proposed polymer matrix as a drug delivery system is safe for human use due to its biodegradability and non-toxicity

Application of an Interactive Whiteboard in the Realization of Entomological Programme Contents

In this paper an analysis of the application of an interactive whiteboard (IWB) in the biology teaching was presented. The types of interactive whiteboards, their advantages and disadvantages, the contextual factors on which the application depends, the positive and negative effects in teaching and learning and pedagogical aspects and strategies for applying interactive whiteboard are described. Also, an example is given how the IWB was applied in the realization of entomological programme content. The role and importance of insects in nature has been shown. It is very important that students understand the role of insect (especially insect pollinators) and their importance in nature. With the help of an IWB, this biology programme content was presented in the form of the concept maps. This allowed students to understand the significance of different entomological concepts and relations among them. Also, an overview of the researches about the uses of an IWB in biology teaching in the realization of various biological programme content was given. It was shown that the IWB contributed to the improving of processes of teaching and learning. It was concluded that IWB are determined as positive teaching tools, and researches show that they could have a positive impact on the motivation, perception, attention, behavior of students and their achievements in teaching biology. Application of IWB can be easily accomplished with little additional training (for teachers and students). In the future, training of teaching staff for the using of IWB in teaching biology is planned

Identification of seed coat phenolic compounds from differently colored pea varieties and characterization of their antioxidant activity

The phenolic composition of seed coats in four differently colored pea varieties (Pisum sativum L.) was investigated using UHPLC-LTQ OrbiTrap MS. The obtained findings revealed that the seed coats of the examined pea genotypes possess a unique phenolic composition compared to previously studied European cultivars. In total, 41 phenolic compounds have been identified. The seed coats of the studied cultivars contained certain amounts of rosmarinic acid, rutin, galangin, morin, naringin, hesperetin and pinocembrin as well as ten flavonol glycosides that had not been reported previously. Additionally, the total phenolic content, antioxidant activity and metal chelating capacity of extracts was determined using Folin-Ciocalteu's method, 2,2-diphenyl-1-picrylhydrazyl radical scavenging assay, ferric ion-reducing capacity and ferrous ion-chelating capacity assay, respectively. Dark colored genotypes MBK 168 and MBK 173 possessed the highest total phenolic contents as well the strongest antioxidant activities. On the other hand, bright colored genotypes MBK 88 and MBK 90 exhibited the strongest metal-chelating capacities. The examined pea seed coats may be considered as important potential contributors to human health due to the presence of bioactive phenolic constituents. In addition, our results could be used as a guideline for breeding new pea cultivars with high antioxidant activities applicable in the formulation of functional food products