Supplementary data for article: Grozdanović, M. M.; Burazer, L. M.; Gavrović-Jankulović, M. Kiwifruit (Actinidia Deliciosa) Extract Shows Potential as a Low-Cost and Efficient Milk-Clotting Agent. International Dairy Journal 2013, 32 (1), 46–52. https://doi.org/10.1016/j.idairyj.2013.03.001

Supplementary material for: [https://doi.org/10.1016/j.idairyj.2013.03.001]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1589

Supplementary data for article: Grozdanovic, M. M.; Drakulić, B. J.; Gavrović-Jankulović, M. Conformational Mobility of Active and E-64-Inhibited Actinidin. Biochimica et Biophysica Acta: General Subjects 2013, 1830 (10), 4790–4799. https://doi.org/10.1016/j.bbagen.2013.06.015

Supplementary material for: [https://doi.org/10.1016/j.bbagen.2013.06.015]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1398

Pregled najčešće korišćenih metoda u karakterizaciji alergena

The characterization of an allergen is a troublesome and difficult process, as it requires both the precise biochemical characterization of a (glyco)protein molecule and the establishment of its susceptibility to IgE antibodies, as they are the main link to histamine release in some hypersensitivity states (type I allergies). As the characterization of an allergen includes molecular weight determination of the allergenic molecule, its structure determination, physicochemical properties, IgE binding properties of the allergen molecule, and its allergenicity, an overal review of which biochemical and immunochemical methods are used in achieving this goal are presented in this paper. The information on the molecular level on the stuctures of allergens indicates that allergens are considerably heterogeneous protein structures, and that there is no particular aminoacid sequence which is responsible for the allergenicity. Therefore, information gained from detailed structural, functional and immunochemical studies of these intriguing molecules, which nowadays modulate a variety of pathophysiological conditions, would greatly improve our understanding of the underlying disease mechanisms, and the way to handle them.Okarakterisati alergen je težak i mukotrpan zadatak, jer zahteva preciznu biohemijsku karakterizaciju (gliko)proteinskog molekula, kao i ustanovljavanje njegove sposobnosti da vezuje IgE, jer je to glavna spona ka oslobađanju histamina u nekim stanjima preosetljivosti (alergije tipa I). Karakterizacija alergena podrazumeva određivanje molekulske mase, određivanje strukture, fizikohemijskih svojstava, IgE vezujućih osobina i njegovu alergenost. Ovaj rad daje pregled koje se biohemijske i imunohemijske metode najčešće koriste radi postizanja tog cilja. Informacije koje su do sada dobijene o strukturi alergena pokazuju da su ovi molekuli izuzetno heterogene strukture i da ne postoji određena aminokiselinska sekvencija koja bi mogla da predvidi alergenost datog molekula. Međutim, informacije dobijene iz detaljnih strukturnih studija ovih molekula će doprineti našem razumevanju patofizioloških procesa koji su u osnovi alergijskih oboljenja i unaprediće način na koji ih tretiramo

Supplementary data for article: Grozdanović, M. M.; Burazer, L. M.; Gavrović-Jankulović, M. Kiwifruit (Actinidia Deliciosa) Extract Shows Potential as a Low-Cost and Efficient Milk-Clotting Agent. International Dairy Journal 2013, 32 (1), 46–52. https://doi.org/10.1016/j.idairyj.2013.03.001

Supplementary material for: [https://doi.org/10.1016/j.idairyj.2013.03.001]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1589

Supplementary data for article: Grozdanovic, M. M.; Drakulić, B. J.; Gavrović-Jankulović, M. Conformational Mobility of Active and E-64-Inhibited Actinidin. Biochimica et Biophysica Acta: General Subjects 2013, 1830 (10), 4790–4799. https://doi.org/10.1016/j.bbagen.2013.06.015

Supplementary material for: [https://doi.org/10.1016/j.bbagen.2013.06.015]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1398

Izolovanje i karakterizacija 68 kD alergena iz ekstrakta kućnih grinja

House dust mites (HDM) represent a major source of allergens, contributing to the increasing incidence of type I hypersensitivity disease worldwide. Over 30 different IgE-binding proteins from the HDM extract were detected. Although group 1 and 2 have been identified as major allergens, due to the safety and efficacy of allergy diagnosis and immunotherapy, there is a need to carefully evaluate the clinical relevance of other allergens present in the HDM extract. In regard to this, a high molecular mass allergen of about 68 kD was purified from the HDM extract using a combination of gel permeation chromatography and reversed-phase chromatography. The IgG and IgE reactivity of the purified protein were preserved during the purification process, as confirmed by Western blot analysis with polyclonal rabbit antibodies and dot blot analysis with a pool of sera from subjects with house dust mite allergy, respectively. In addition, the IgE reactivity was confirmed using ELISA testing with nine patient sera. The biological potency of the 68 kD allergen was confirmed by skin prick testing in five allergic subjects, suggesting that the high molecular mass allergen is a good candidate for component-resolved diagnosis of house dust mite allergy and eventual therapeutic treatment.Grinje iz kućne prašine predstavljaju jedan od glavnih izvora alergena koji su u značajnoj meri doprineli porastu prvog tipa preosetljivosti. Preko 30 IgE-vezujućih proteina iz kućne prašine je detektovano do danas. Alergeni grupe 1 i 2 označeni su kao glavni alergeni kućne prašine. Međutim, da bi se poboljšala sigurnost i efikasnost dijagnoze i terapije alergijskih oboljenja izazvanih grinjama iz kućne prašine, neophodno je odrediti klinički značaj svih alergena iz ovog alergenskog izvora. U ovom radu izolovan je alergen visoke molekulske mase od 68 kD iz ekstrakta kućne prašine kombinovanjem gel-permeacione hromatografije i reversno-fazne hromatografije. IgG i IgE reaktivnost prečišćenog proteina je proverena u 'Western blot'-u i 'dot blot'-u sa poliklonskim zečijim antitelima na ekstrakt kućne prašine i 'pool'-om seruma osoba alergičnih na kućnu prašinu, redom. 64 % pacijenata je pokazalo IgE reaktivnost na prečišćeni protein u ELISA testu. Biološka reaktivnost prečišćenog alergena je potvrđena u kožnim probama na pet pacijenata, ukazujući da je prečišćen alergen dobar kandidat za dijagnozu alergije na kućnu prašinu pojedinačnim komponentama i eventualni terapeutski tretman

Parcijalna karakterizacija antibakterijskog peptida koji proizvodi probiotska bakterija Lactobacillus plantarum G2

The aim of this study was the partial characterization of the antimicrobial peptide bacteriocin G, produced by probiotic bacteria Lactobacillus plantarum G(2), which was isolated from a clinical sample of a healthy person. Antimicrobial substance was secreted in the supernatant of an L. plantarum G(2) culture, and showed a diverse spectrum of antimicrobial activity of all the tested strains of the genera Lactobacillus and the pathogenic bacteria Staphylococcus aureus and Salmonella abony. Isoclectric focusing revealed that bacteriocin G(2) is a cationic peptide (pI about 10) with a molecular mass of 2.2 kDa according to tricine sodium dodecyl sulphate polyacrylamide gel electrophoresis, SDS-PAGE. The antimicrobial activity of bactcriocin G(2) was diminished by the proteolytic action of trypsin and proteinase K. Bacteriocin G(2) preserved its biological activity in the temperature range 40-60 degrees C (15 min), which was lost at 80 degrees C. Bacteriocin G(2) was stable in the pH range 2-9, while treatment with 1 % Tween 80 and 1 % urea resulted in increased antimicrobial activity. The probiotic strain L. plantarum G(2) produces the antimicrobial substance proteinaceous in nature with bacteriocin characteristics. Bacteriocin production is one of the key properties of probiotic bacteria with clinical potential as anti-infective agents, which will increase the likelihood of its in vivo efficacy.Cilj ovog istraživanja je bila parcijalna karakterizacija antimikrobnog peptida iz probiotske bakterija Lactobacillus plantarum G2 izolovane iz kliničkog uzorka zdrave osobe. Antimikrobno jedinjenje iz L. plantarum G2, označeno kao bakteriocin G2, dobijeno iz supernatanta bakterijske kulture, pokazalo je širok spektar antimikrobne aktivnosti, inhibirajići rast svih ispitivanh vrsta roda Lactobacillus, kao i patogenih bakterija Staphylococcus aureus i Salmonella abony. Bakteriocin G2 je osetljiv na proteolitičko dejstvo tripsina i proteinaze K. Antimikrobna aktivnost je stabilna u opsegu 40-60°C (15 min), ali se gubi na temperaturi od 80°C. Ustanovljeno je da je bakteriocin stabilan na pH vrednostima između 2 i 9. Dejstvo Tween-a 80 i uree je dovelo do povećane inhibitorne aktivnosti. Prema IEF, bakteriocin G2 je katjonski protein, sa pI vrednošću oko 10, a molekulaska masa određena na osnovu tricin-SDS-PAGE je 2,2 kDa. Probiotski soj L plantarum G2 produkuje antimikrobno jedinjenje proteinske strukture sa karakteristikama bakteriocina. Sinteza bakteriocina je jedna od ključnih osobina probiotskih bakterija koje imaju klinički potencijal kao antiinfektivni agensi, jer značajno povećava verovatnoću njihove in vivo efikasnosti

La(OH)3 Multi-Walled Carbon Nanotube/Carbon Paste-Based Sensing Approach for the Detection of Uric Acid—A Product of Environmentally Stressed Cells

This paper aims to develop an amperometric, non-enzymatic sensor for detecting and quantifying UA as an alert signal induced by allergens with protease activity in human cell lines (HEK293 and HeLa). Uric acid (UA) has been classified as a damage-associated molecular pattern (DAMP) molecule that serves a physiological purpose inside the cell, while outside the cell it can be an indicator of cell damage. Cell damage or stress can be caused by different health problems or by environmental irritants, such as allergens. We can act and prevent the events that generate stress by determining the extent to which cells are under stress. Amperometric calibration measurements were performed with a carbon paste electrode modified with La(OH)3@MWCNT, at the potential of 0.3 V. The calibration curve was constructed in a linear operating range from 0.67 μM to 121 μM UA. The proposed sensor displayed good reproducibility with an RSD of 3.65% calculated for five subsequent measurements, and a low detection limit of 64.28 nM, determined using the 3 S/m method. Interference studies and the real sample analysis of allergen-treated cell lines proved that the proposed sensing platform possesses excellent sensitivity, reproducibility, and stability. Therefore, it can potentially be used to evaluate stress factors in medical research and clinical practice

Supplementary material for the article: Stojsavljević, A.; Rovčanin, B.; Krstić; Jagodić, J.; Borković-Mitić, S.; Paunović, I.; Živaljević, V.; Mitić, B.; Gavrović-Jankulović, M.; Manojlović, D. Cadmium as Main Endocrine Disruptor in Papillary Thyroid Carcinoma and the Significance of Cd/Se Ratio for Thyroid Tissue Pathophysiology. Journal of Trace Elements in Medicine and Biology 2019, 55, 190–195. https://doi.org/10.1016/j.jtemb.2019.06.009

Supplementary material for: [https://doi.org/10.1016/j.jtemb.2019.06.009]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/3293

Supplementary material for the article: Stojsavljević, A.; Rovčanin, B.; Krstić; Jagodić, J.; Borković-Mitić, S.; Paunović, I.; Živaljević, V.; Mitić, B.; Gavrović-Jankulović, M.; Manojlović, D. Cadmium as Main Endocrine Disruptor in Papillary Thyroid Carcinoma and the Significance of Cd/Se Ratio for Thyroid Tissue Pathophysiology. Journal of Trace Elements in Medicine and Biology 2019, 55, 190–195. https://doi.org/10.1016/j.jtemb.2019.06.009

Supplementary material for: [https://doi.org/10.1016/j.jtemb.2019.06.009]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/3293