TY  - JOUR
AU  - Mladenović Stokanić, Maja
AU  - Simović, Ana
AU  - Jovanović, Vesna B.
AU  - Radomirović, Mirjana
AU  - Udovićki, Božidar
AU  - Krstić Ristivojević, Maja
AU  - Đukić, Teodora
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Sabljić, Ljiljana
AU  - Lukić, Ivana
AU  - Kovačević, Ana
AU  - Cujić, Danica
AU  - Gnjatović, Marija
AU  - Smiljanić, Katarina
AU  - Stojadinović, Marija
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojanović, Marijana
AU  - Rajković, Andreja
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6436
AB  - In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species
VL  - 25
IS  - 1
SP  - 333
DO  - 10.3390/ijms25010333
ER  - 

@article{
author = "Mladenović Stokanić, Maja and Simović, Ana and Jovanović, Vesna B. and Radomirović, Mirjana and Udovićki, Božidar and Krstić Ristivojević, Maja and Đukić, Teodora and Vasović, Tamara and Aćimović, Jelena and Sabljić, Ljiljana and Lukić, Ivana and Kovačević, Ana and Cujić, Danica and Gnjatović, Marija and Smiljanić, Katarina and Stojadinović, Marija and Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojanović, Marijana and Rajković, Andreja and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species",
volume = "25",
number = "1",
pages = "333",
doi = "10.3390/ijms25010333"
}

Mladenović Stokanić, M., Simović, A., Jovanović, V. B., Radomirović, M., Udovićki, B., Krstić Ristivojević, M., Đukić, T., Vasović, T., Aćimović, J., Sabljić, L., Lukić, I., Kovačević, A., Cujić, D., Gnjatović, M., Smiljanić, K., Stojadinović, M., Radosavljević, J., Stanić-Vučinić, D., Stojanović, M., Rajković, A.,& Ćirković-Veličković, T.. (2023). Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences
MDPI., 25(1), 333.
https://doi.org/10.3390/ijms25010333

Mladenović Stokanić M, Simović A, Jovanović VB, Radomirović M, Udovićki B, Krstić Ristivojević M, Đukić T, Vasović T, Aćimović J, Sabljić L, Lukić I, Kovačević A, Cujić D, Gnjatović M, Smiljanić K, Stojadinović M, Radosavljević J, Stanić-Vučinić D, Stojanović M, Rajković A, Ćirković-Veličković T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences. 2023;25(1):333.
doi:10.3390/ijms25010333 .

Mladenović Stokanić, Maja, Simović, Ana, Jovanović, Vesna B., Radomirović, Mirjana, Udovićki, Božidar, Krstić Ristivojević, Maja, Đukić, Teodora, Vasović, Tamara, Aćimović, Jelena, Sabljić, Ljiljana, Lukić, Ivana, Kovačević, Ana, Cujić, Danica, Gnjatović, Marija, Smiljanić, Katarina, Stojadinović, Marija, Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojanović, Marijana, Rajković, Andreja, Ćirković-Veličković, Tanja, "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species" in International Journal of Molecular Sciences, 25, no. 1 (2023):333,
https://doi.org/10.3390/ijms25010333 . .

TY  - JOUR
AU  - Slavić, Marinela Šokarda
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Stanisavljević, Nemanja S.
AU  - Gardijan, Lazar
AU  - Božić, Nataša
AU  - Vujčić, Zoran
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6351
AB  - α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60–80 °C and is active and stable over a wide pH range (4.0–9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications
VL  - 249
SP  - 126055
DO  - 10.1016/j.ijbiomac.2023.126055
ER  - 

@article{
author = "Slavić, Marinela Šokarda and Kojić, Milan and Margetić, Aleksandra and Stanisavljević, Nemanja S. and Gardijan, Lazar and Božić, Nataša and Vujčić, Zoran",
year = "2023",
abstract = "α-Amylase from the thermophilic bacterial strain Anoxybacillus vranjensis ST4 (AVA) was cloned into the pMALc5HisEk expression vector and successfully expressed and purified from the Escherichia coli ER2523 host strain. AVA belongs to the GH13_5 subfamily of glycoside hydrolases and has 7 conserved sequence regions (CSRs) distributed in three distinct domains (A, B, C). In addition, there is a starch binding domain (SBD) from the CBM20 family of carbohydrate binding modules (CBMs). AVA is a monomer of 66 kDa that achieves maximum activity at 60–80 °C and is active and stable over a wide pH range (4.0–9.0). AVA retained 50 % of its activity after 31 h of incubation at 60 °C and was resistant to a large number of denaturing agents. It hydrolyzed starch granules very efficiently, releasing maltose, maltotriose and maltopentaose as the main products. The hydrolysis rates of raw corn, wheat, horseradish, and potato starch, at a concentration of 10 %, were 87.8, 85.9, 93.0, and 58 %, respectively, at pH 8.5 over a 3 h period. This study showed that the high level of expression as well as the properties of this highly stable and versatile enzyme show all the prerequisites for successful application in industry.",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications",
volume = "249",
pages = "126055",
doi = "10.1016/j.ijbiomac.2023.126055"
}

Slavić, M. Š., Kojić, M., Margetić, A., Stanisavljević, N. S., Gardijan, L., Božić, N.,& Vujčić, Z.. (2023). Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications. in International Journal of Biological Macromolecules
Elsevier., 249, 126055.
https://doi.org/10.1016/j.ijbiomac.2023.126055

Slavić MŠ, Kojić M, Margetić A, Stanisavljević NS, Gardijan L, Božić N, Vujčić Z. Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications. in International Journal of Biological Macromolecules. 2023;249:126055.
doi:10.1016/j.ijbiomac.2023.126055 .

Slavić, Marinela Šokarda, Kojić, Milan, Margetić, Aleksandra, Stanisavljević, Nemanja S., Gardijan, Lazar, Božić, Nataša, Vujčić, Zoran, "Highly stable and versatile α-amylase from Anoxybacillus vranjensis ST4 suitable for various applications" in International Journal of Biological Macromolecules, 249 (2023):126055,
https://doi.org/10.1016/j.ijbiomac.2023.126055 . .

TY  - JOUR
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Trifunović, Sara
AU  - Krstić-Ristivojević, Maja
AU  - Aćimović, Milica G.
AU  - Stanković Jeremić, Jovana
AU  - Lončar, Biljana
AU  - Tešević, Vele
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6203
AB  - As byproducts of essential oil distillation, hydrolates are used in natural cosmetics/biomedicine due to their beneficial skin effects. However, data on their safety with relevant biological targets, such as human skin cells, are scarce. Therefore, we have tested nine hydrolates from the Lamiaceae family with skin fibroblasts that are responsible for extracellular collagenous matrix builds. Thyme, oregano, and winter savoury hydrolates showed several times higher total phenolics, which correlated strongly with their radical scavenging and antioxidative capacity; there was no correlation between their viability profiles and the reducing sugar levels. No proteins/peptides were detected. All hydrolates appeared safe for prolonged skin exposure except for 10-fold diluted lavender, which showed cytotoxicity (~20%), as well as rosemary and lavandin (~10%) using viability, DNA synthesis, and cell count testing. Clary sage, oregano, lemon balm, and thyme hydrolates (10-fold diluted) increased fibroblast viability and/or proliferation by 10–30% compared with the control, while their viability remained unaffected by Mentha and winter savoury. In line with the STITCH database, increased viability could be attributed to thymol presence in oregano and thyme hydrolates in lemon balm, which is most likely attributable to neral and geranial. The proliferative effect of clary sage could be supported by alpha-terpineol, not linalool. The major volatile organic compounds (VOCs) associated with cytotoxic effects on fibroblasts were borneol, 1,8-cineole, and terpinene-4-ol. Further research with pure compounds is warranted to confirm the roles of VOCs in the observed effects that are relevant to cosmetic and wound healing aspects.
PB  - MDPI
T2  - Antioxidants
T1  - Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts
VL  - 12
IS  - 11
DO  - 10.3390/antiox12111988
ER  - 

@article{
author = "Smiljanić, Katarina and Prodić, Ivana and Trifunović, Sara and Krstić-Ristivojević, Maja and Aćimović, Milica G. and Stanković Jeremić, Jovana and Lončar, Biljana and Tešević, Vele",
year = "2023",
abstract = "As byproducts of essential oil distillation, hydrolates are used in natural cosmetics/biomedicine due to their beneficial skin effects. However, data on their safety with relevant biological targets, such as human skin cells, are scarce. Therefore, we have tested nine hydrolates from the Lamiaceae family with skin fibroblasts that are responsible for extracellular collagenous matrix builds. Thyme, oregano, and winter savoury hydrolates showed several times higher total phenolics, which correlated strongly with their radical scavenging and antioxidative capacity; there was no correlation between their viability profiles and the reducing sugar levels. No proteins/peptides were detected. All hydrolates appeared safe for prolonged skin exposure except for 10-fold diluted lavender, which showed cytotoxicity (~20%), as well as rosemary and lavandin (~10%) using viability, DNA synthesis, and cell count testing. Clary sage, oregano, lemon balm, and thyme hydrolates (10-fold diluted) increased fibroblast viability and/or proliferation by 10–30% compared with the control, while their viability remained unaffected by Mentha and winter savoury. In line with the STITCH database, increased viability could be attributed to thymol presence in oregano and thyme hydrolates in lemon balm, which is most likely attributable to neral and geranial. The proliferative effect of clary sage could be supported by alpha-terpineol, not linalool. The major volatile organic compounds (VOCs) associated with cytotoxic effects on fibroblasts were borneol, 1,8-cineole, and terpinene-4-ol. Further research with pure compounds is warranted to confirm the roles of VOCs in the observed effects that are relevant to cosmetic and wound healing aspects.",
publisher = "MDPI",
journal = "Antioxidants",
title = "Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts",
volume = "12",
number = "11",
doi = "10.3390/antiox12111988"
}

Smiljanić, K., Prodić, I., Trifunović, S., Krstić-Ristivojević, M., Aćimović, M. G., Stanković Jeremić, J., Lončar, B.,& Tešević, V.. (2023). Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts. in Antioxidants
MDPI., 12(11).
https://doi.org/10.3390/antiox12111988

Smiljanić K, Prodić I, Trifunović S, Krstić-Ristivojević M, Aćimović MG, Stanković Jeremić J, Lončar B, Tešević V. Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts. in Antioxidants. 2023;12(11).
doi:10.3390/antiox12111988 .

Smiljanić, Katarina, Prodić, Ivana, Trifunović, Sara, Krstić-Ristivojević, Maja, Aćimović, Milica G., Stanković Jeremić, Jovana, Lončar, Biljana, Tešević, Vele, "Multistep Approach Points to Compounds Responsible for the Biological Activity and Safety of Hydrolates from Nine Lamiaceae Medicinal Plants on Human Skin Fibroblasts" in Antioxidants, 12, no. 11 (2023),
https://doi.org/10.3390/antiox12111988 . .

TY  - JOUR
AU  - Šokarda Slavić, Marinela
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Ristović, Marina
AU  - Pavlović, Marija
AU  - Nikolić, Stefan
AU  - Vujčić, Zoran
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5992
AB  - The combination ofb-oligosaccharides from enzymatically hydrolysed barleyb-glucan has attracted inter-est recently due to its positive effects on human health. This study aimed to assess the impact of theendo-b-1,3-1,4-glucanase enzyme fromBacillus  subtilis168 on improving the nutritional and bioactiveproperties of barleyb-glucan. A new procedure for the isolation ofb-glucan was developed, at a lowertemperature (45°C), enabling purity from starch contamination, without affecting the yield (6 gb-glucanfrom 100 g of barley flour). The endo-b-1,3-1,4-glucanase is cloned intoE. colipQE_Ek enables the highproduction and purification (82% yield, 1.8 mg mL 1and 440 U mg 1) of an enzyme identical to thenatural one (25.5 kDa). The enzymatic reaction showed high efficiency ofb-glucan degradation by recom-binant enzyme, giving a mixture of products (of which 3-O-b-cellobiosyl-D-glucose and 3-O-b-cellotriosyl-D-glucose are the most abundant), the reduction of viscosity (17%) and increase in antioxidant capacitiesby 15.2%, 30.9% and 44.0% assessed by ABTS, DPPH and ORAC, respectively. These results indicatethe possible application of endo-b-1,3-1,4-glucanase enzyme in improving the properties of barleyb-glucan used as functional foods.
PB  - Wiley
T2  - International Journal of Food Science and Technology
T1  - Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase
VL  - 58
IS  - 12
SP  - 6825
EP  - 6835
DO  - 10.1111/ijfs.16647
ER  - 

@article{
author = "Šokarda Slavić, Marinela and Kojić, Milan and Margetić, Aleksandra and Ristović, Marina and Pavlović, Marija and Nikolić, Stefan and Vujčić, Zoran",
year = "2023",
abstract = "The combination ofb-oligosaccharides from enzymatically hydrolysed barleyb-glucan has attracted inter-est recently due to its positive effects on human health. This study aimed to assess the impact of theendo-b-1,3-1,4-glucanase enzyme fromBacillus  subtilis168 on improving the nutritional and bioactiveproperties of barleyb-glucan. A new procedure for the isolation ofb-glucan was developed, at a lowertemperature (45°C), enabling purity from starch contamination, without affecting the yield (6 gb-glucanfrom 100 g of barley flour). The endo-b-1,3-1,4-glucanase is cloned intoE. colipQE_Ek enables the highproduction and purification (82% yield, 1.8 mg mL 1and 440 U mg 1) of an enzyme identical to thenatural one (25.5 kDa). The enzymatic reaction showed high efficiency ofb-glucan degradation by recom-binant enzyme, giving a mixture of products (of which 3-O-b-cellobiosyl-D-glucose and 3-O-b-cellotriosyl-D-glucose are the most abundant), the reduction of viscosity (17%) and increase in antioxidant capacitiesby 15.2%, 30.9% and 44.0% assessed by ABTS, DPPH and ORAC, respectively. These results indicatethe possible application of endo-b-1,3-1,4-glucanase enzyme in improving the properties of barleyb-glucan used as functional foods.",
publisher = "Wiley",
journal = "International Journal of Food Science and Technology",
title = "Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase",
volume = "58",
number = "12",
pages = "6825-6835",
doi = "10.1111/ijfs.16647"
}

Šokarda Slavić, M., Kojić, M., Margetić, A., Ristović, M., Pavlović, M., Nikolić, S.,& Vujčić, Z.. (2023). Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase. in International Journal of Food Science and Technology
Wiley., 58(12), 6825-6835.
https://doi.org/10.1111/ijfs.16647

Šokarda Slavić M, Kojić M, Margetić A, Ristović M, Pavlović M, Nikolić S, Vujčić Z. Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase. in International Journal of Food Science and Technology. 2023;58(12):6825-6835.
doi:10.1111/ijfs.16647 .

Šokarda Slavić, Marinela, Kojić, Milan, Margetić, Aleksandra, Ristović, Marina, Pavlović, Marija, Nikolić, Stefan, Vujčić, Zoran, "Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase" in International Journal of Food Science and Technology, 58, no. 12 (2023):6825-6835,
https://doi.org/10.1111/ijfs.16647 . .

TY  - JOUR
AU  - Šokarda Slavić, Marinela
AU  - Kojić, Milan
AU  - Margetić, Aleksandra
AU  - Ristović, Marina
AU  - Pavlović, Marija
AU  - Nikolić, Stefan
AU  - Vujčić, Zoran
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6008
AB  - The combination ofb-oligosaccharides from enzymatically hydrolysed barleyb-glucan has attracted inter-est recently due to its positive effects on human health. This study aimed to assess the impact of theendo-b-1,3-1,4-glucanase enzyme fromBacillus  subtilis168 on improving the nutritional and bioactiveproperties of barleyb-glucan. A new procedure for the isolation ofb-glucan was developed, at a lowertemperature (45°C), enabling purity from starch contamination, without affecting the yield (6 gb-glucanfrom 100 g of barley flour). The endo-b-1,3-1,4-glucanase is cloned intoE. colipQE_Ek enables the highproduction and purification (82% yield, 1.8 mg mL 1and 440 U mg 1) of an enzyme identical to thenatural one (25.5 kDa). The enzymatic reaction showed high efficiency ofb-glucan degradation by recom-binant enzyme, giving a mixture of products (of which 3-O-b-cellobiosyl-D-glucose and 3-O-b-cellotriosyl-D-glucose are the most abundant), the reduction of viscosity (17%) and increase in antioxidant capacitiesby 15.2%, 30.9% and 44.0% assessed by ABTS, DPPH and ORAC, respectively. These results indicatethe possible application of endo-b-1,3-1,4-glucanase enzyme in improving the properties of barleyb-glucan used as functional foods.
PB  - Wiley
T2  - International Journal of Food Science and Technology
T1  - Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase
VL  - 58
IS  - 12
SP  - 6825
EP  - 6835
DO  - 10.1111/ijfs.16647
ER  - 

@article{
author = "Šokarda Slavić, Marinela and Kojić, Milan and Margetić, Aleksandra and Ristović, Marina and Pavlović, Marija and Nikolić, Stefan and Vujčić, Zoran",
year = "2023",
abstract = "The combination ofb-oligosaccharides from enzymatically hydrolysed barleyb-glucan has attracted inter-est recently due to its positive effects on human health. This study aimed to assess the impact of theendo-b-1,3-1,4-glucanase enzyme fromBacillus  subtilis168 on improving the nutritional and bioactiveproperties of barleyb-glucan. A new procedure for the isolation ofb-glucan was developed, at a lowertemperature (45°C), enabling purity from starch contamination, without affecting the yield (6 gb-glucanfrom 100 g of barley flour). The endo-b-1,3-1,4-glucanase is cloned intoE. colipQE_Ek enables the highproduction and purification (82% yield, 1.8 mg mL 1and 440 U mg 1) of an enzyme identical to thenatural one (25.5 kDa). The enzymatic reaction showed high efficiency ofb-glucan degradation by recom-binant enzyme, giving a mixture of products (of which 3-O-b-cellobiosyl-D-glucose and 3-O-b-cellotriosyl-D-glucose are the most abundant), the reduction of viscosity (17%) and increase in antioxidant capacitiesby 15.2%, 30.9% and 44.0% assessed by ABTS, DPPH and ORAC, respectively. These results indicatethe possible application of endo-b-1,3-1,4-glucanase enzyme in improving the properties of barleyb-glucan used as functional foods.",
publisher = "Wiley",
journal = "International Journal of Food Science and Technology",
title = "Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase",
volume = "58",
number = "12",
pages = "6825-6835",
doi = "10.1111/ijfs.16647"
}

Šokarda Slavić, M., Kojić, M., Margetić, A., Ristović, M., Pavlović, M., Nikolić, S.,& Vujčić, Z.. (2023). Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase. in International Journal of Food Science and Technology
Wiley., 58(12), 6825-6835.
https://doi.org/10.1111/ijfs.16647

Šokarda Slavić M, Kojić M, Margetić A, Ristović M, Pavlović M, Nikolić S, Vujčić Z. Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase. in International Journal of Food Science and Technology. 2023;58(12):6825-6835.
doi:10.1111/ijfs.16647 .

Šokarda Slavić, Marinela, Kojić, Milan, Margetić, Aleksandra, Ristović, Marina, Pavlović, Marija, Nikolić, Stefan, Vujčić, Zoran, "Improvement of nutritional and bioactive properties of barleyb-glucan-based food products usingBacillus subtilis168endo-b-1,3-1,4-glucanase" in International Journal of Food Science and Technology, 58, no. 12 (2023):6825-6835,
https://doi.org/10.1111/ijfs.16647 . .

TY  - JOUR
AU  - Milovanović, Vladimir
AU  - Minić, Rajna
AU  - Vakić, Jelena
AU  - Ivanović, Saša
AU  - Čupić, Vitomir
AU  - Borozan, Sunćica
AU  - Nešić, Andrijana N.
AU  - Živković, Irena
PY  - 2021
UR  - https://www.sciencedirect.com/science/article/pii/S0041010121000362
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4490
AB  - The MTT assay is routinely used to detect the activity of living cells. While working with Vipera ammodytes venom we detected the reduction of MTT without the presence of cells, in a concentration-dependent manner. By combining non-reducing PAGE, L-amino acid oxidase (LAAO) assays, and standard MTT assays, we established and confirmed that venom MTT reduction is catalyzed by only one enzyme, the LAAO. Even though it was previously known that the dimeric and tetrameric forms of LAAO are active, we conclude that the enzyme is also active in the monomeric form. Our results have led to the definition of a new MTT assay in a microtiter plate for in vitro testing of svLAAO activity i.e. from the venom of the V. ammodytes snake. Potentially, this method can be used for testing hemorrhagic venoms of other snakes as well as the LAAO neutralization capability of appropriate antivenoms.
PB  - Elsevier
T2  - Toxicon
T2  - ToxiconToxicon
T1  - MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation
VL  - 192
SP  - 57
EP  - 65
DO  - 10.1016/j.toxicon.2021.01.012
ER  - 

@article{
author = "Milovanović, Vladimir and Minić, Rajna and Vakić, Jelena and Ivanović, Saša and Čupić, Vitomir and Borozan, Sunćica and Nešić, Andrijana N. and Živković, Irena",
year = "2021",
abstract = "The MTT assay is routinely used to detect the activity of living cells. While working with Vipera ammodytes venom we detected the reduction of MTT without the presence of cells, in a concentration-dependent manner. By combining non-reducing PAGE, L-amino acid oxidase (LAAO) assays, and standard MTT assays, we established and confirmed that venom MTT reduction is catalyzed by only one enzyme, the LAAO. Even though it was previously known that the dimeric and tetrameric forms of LAAO are active, we conclude that the enzyme is also active in the monomeric form. Our results have led to the definition of a new MTT assay in a microtiter plate for in vitro testing of svLAAO activity i.e. from the venom of the V. ammodytes snake. Potentially, this method can be used for testing hemorrhagic venoms of other snakes as well as the LAAO neutralization capability of appropriate antivenoms.",
publisher = "Elsevier",
journal = "Toxicon, ToxiconToxicon",
title = "MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation",
volume = "192",
pages = "57-65",
doi = "10.1016/j.toxicon.2021.01.012"
}

Milovanović, V., Minić, R., Vakić, J., Ivanović, S., Čupić, V., Borozan, S., Nešić, A. N.,& Živković, I.. (2021). MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation. in Toxicon
Elsevier., 192, 57-65.
https://doi.org/10.1016/j.toxicon.2021.01.012

Milovanović V, Minić R, Vakić J, Ivanović S, Čupić V, Borozan S, Nešić AN, Živković I. MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation. in Toxicon. 2021;192:57-65.
doi:10.1016/j.toxicon.2021.01.012 .

Milovanović, Vladimir, Minić, Rajna, Vakić, Jelena, Ivanović, Saša, Čupić, Vitomir, Borozan, Sunćica, Nešić, Andrijana N., Živković, Irena, "MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation" in Toxicon, 192 (2021):57-65,
https://doi.org/10.1016/j.toxicon.2021.01.012 . .

TY  - JOUR
AU  - Protić-Rosić, Isidora
AU  - Nešić, Andrijana N.
AU  - Lukić, Ivana
AU  - Miljković, Radmila
AU  - Popović, Dragan M.
AU  - Atanasković-Marković, Marina
AU  - Stojanović, Marijana M.
AU  - Gavrović-Jankulović, Marija
PY  - 2021
UR  - https://www.sciencedirect.com/science/article/pii/S0161589021001905
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4604
AB  - Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.
PB  - Elsevier
T2  - Molecular Immunology
T2  - Molecular ImmunologyMolecular Immunology
T1  - Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion
VL  - 138
SP  - 58
EP  - 67
DO  - 10.1016/j.molimm.2021.06.015
ER  - 

@article{
author = "Protić-Rosić, Isidora and Nešić, Andrijana N. and Lukić, Ivana and Miljković, Radmila and Popović, Dragan M. and Atanasković-Marković, Marina and Stojanović, Marijana M. and Gavrović-Jankulović, Marija",
year = "2021",
abstract = "Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.",
publisher = "Elsevier",
journal = "Molecular Immunology, Molecular ImmunologyMolecular Immunology",
title = "Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion",
volume = "138",
pages = "58-67",
doi = "10.1016/j.molimm.2021.06.015"
}

Protić-Rosić, I., Nešić, A. N., Lukić, I., Miljković, R., Popović, D. M., Atanasković-Marković, M., Stojanović, M. M.,& Gavrović-Jankulović, M.. (2021). Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion. in Molecular Immunology
Elsevier., 138, 58-67.
https://doi.org/10.1016/j.molimm.2021.06.015

Protić-Rosić I, Nešić AN, Lukić I, Miljković R, Popović DM, Atanasković-Marković M, Stojanović MM, Gavrović-Jankulović M. Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion. in Molecular Immunology. 2021;138:58-67.
doi:10.1016/j.molimm.2021.06.015 .

Protić-Rosić, Isidora, Nešić, Andrijana N., Lukić, Ivana, Miljković, Radmila, Popović, Dragan M., Atanasković-Marković, Marina, Stojanović, Marijana M., Gavrović-Jankulović, Marija, "Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion" in Molecular Immunology, 138 (2021):58-67,
https://doi.org/10.1016/j.molimm.2021.06.015 . .

TY  - JOUR
AU  - Lopandić, Zorana
AU  - Dragačević, Luka
AU  - Popović, Dragan M.
AU  - Anđelković, Uroš
AU  - Minić, Rajna
AU  - Gavrović-Jankulović, Marija
PY  - 2021
UR  - https://www.mdpi.com/2218-273X/11/2/180
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4489
AB  - Fluorescently labeled lectins are useful tools for in vivo and in vitro studies of the structure and function of tissues and various pathogens such as viruses, bacteria, and fungi. For the evaluation of high-mannose glycans present on various glycoproteins, a three-dimensional (3D) model of the chimera was designed from the crystal structures of recombinant banana lectin (BanLec, Protein Data Bank entry (PDB): 5EXG) and an enhanced green fluorescent protein (eGFP, PDB 4EUL) by applying molecular modeling and molecular mechanics and expressed in Escherichia coli. BanLec-eGFP, produced as a soluble cytosolic protein of about 42 kDa, revealed β-sheets (41%) as the predominant secondary structures, with the emission peak maximum detected at 509 nm (excitation wavelength 488 nm). More than 65% of the primary structure was confirmed by mass spectrometry. Competitive BanLec-eGFP binding to high mannose glycans of the influenza vaccine (Vaxigrip®) was shown in a fluorescence-linked lectin sorbent assay (FLLSA) with monosaccharides (mannose and glucose) and wild type BanLec and H84T BanLec mutant. BanLec-eGFP exhibited binding to mannose residues on different strains of Salmonella in flow cytometry, with especially pronounced binding to a Salmonella Typhi clinical isolate. BanLec-eGFP can be a useful tool for screening high-mannose glycosylation sites on different microorganisms.
PB  - MDPI
T2  - Biomolecules
T2  - Biomolecules
T1  - BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms
VL  - 11
IS  - 2
SP  - 180
DO  - 10.3390/biom11020180
ER  - 

@article{
author = "Lopandić, Zorana and Dragačević, Luka and Popović, Dragan M. and Anđelković, Uroš and Minić, Rajna and Gavrović-Jankulović, Marija",
year = "2021",
abstract = "Fluorescently labeled lectins are useful tools for in vivo and in vitro studies of the structure and function of tissues and various pathogens such as viruses, bacteria, and fungi. For the evaluation of high-mannose glycans present on various glycoproteins, a three-dimensional (3D) model of the chimera was designed from the crystal structures of recombinant banana lectin (BanLec, Protein Data Bank entry (PDB): 5EXG) and an enhanced green fluorescent protein (eGFP, PDB 4EUL) by applying molecular modeling and molecular mechanics and expressed in Escherichia coli. BanLec-eGFP, produced as a soluble cytosolic protein of about 42 kDa, revealed β-sheets (41%) as the predominant secondary structures, with the emission peak maximum detected at 509 nm (excitation wavelength 488 nm). More than 65% of the primary structure was confirmed by mass spectrometry. Competitive BanLec-eGFP binding to high mannose glycans of the influenza vaccine (Vaxigrip®) was shown in a fluorescence-linked lectin sorbent assay (FLLSA) with monosaccharides (mannose and glucose) and wild type BanLec and H84T BanLec mutant. BanLec-eGFP exhibited binding to mannose residues on different strains of Salmonella in flow cytometry, with especially pronounced binding to a Salmonella Typhi clinical isolate. BanLec-eGFP can be a useful tool for screening high-mannose glycosylation sites on different microorganisms.",
publisher = "MDPI",
journal = "Biomolecules, Biomolecules",
title = "BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms",
volume = "11",
number = "2",
pages = "180",
doi = "10.3390/biom11020180"
}

Lopandić, Z., Dragačević, L., Popović, D. M., Anđelković, U., Minić, R.,& Gavrović-Jankulović, M.. (2021). BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms. in Biomolecules
MDPI., 11(2), 180.
https://doi.org/10.3390/biom11020180

Lopandić Z, Dragačević L, Popović DM, Anđelković U, Minić R, Gavrović-Jankulović M. BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms. in Biomolecules. 2021;11(2):180.
doi:10.3390/biom11020180 .

Lopandić, Zorana, Dragačević, Luka, Popović, Dragan M., Anđelković, Uroš, Minić, Rajna, Gavrović-Jankulović, Marija, "BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms" in Biomolecules, 11, no. 2 (2021):180,
https://doi.org/10.3390/biom11020180 . .

TY  - JOUR
AU  - Stojanović, Marijana M.
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Kovačević, Ana
AU  - Miljković, Radmila
AU  - Tobias, Joshua
AU  - Schabussova, Irma
AU  - Zlatović, Mario
AU  - Barisani-Asenbauer, Talin
AU  - Wiedermann, Ursula
AU  - Inic-Kanada, Aleksandra
PY  - 2020
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4292
AB  - Vaccines can have heterologous effects on the immune system, i.e., effects other than triggering an immune response against the disease targeted by the vaccine. We investigated whether monoclonal antibodies (mAbs) specific for tetanus could cross-react with Chlamydia and confer heterologous protection against chlamydial infection. The capability of two tetanus-specific mAbs, namely mAb26 and mAb51, to prevent chlamydial infection has been assessed: (i) in vitro, by performing a neutralization assay using human conjunctival epithelial (HCjE) cells infected with Chlamydia trachomatis serovar B, and (ii) in vivo, by using a guinea pig model of Chlamydiacaviae-induced inclusion conjunctivitis. The mAb26 has been superior in comparison with mAb51 in the prevention of chlamydial infection in HCjE cells. The mAb26 has conferred ≈40% inhibition of the infection, compared to less than 5% inhibition in the presence of the mAb51. In vivo, mAb26 significantly diminished ocular pathology intensity in guinea pigs infected with C. caviae compared to either the mAb51-treated or sham-treated guinea pigs. Our data provide insights that tetanus immunization generates antibodies which induce heterologous chlamydial immunity and promote protection beyond the intended target pathogen.
T2  - Vaccines
T1  - Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia
VL  - 8
IS  - 4
SP  - 719
DO  - 10.3390/vaccines8040719
ER  - 

@article{
author = "Stojanović, Marijana M. and Lukić, Ivana and Marinković, Emilija and Kovačević, Ana and Miljković, Radmila and Tobias, Joshua and Schabussova, Irma and Zlatović, Mario and Barisani-Asenbauer, Talin and Wiedermann, Ursula and Inic-Kanada, Aleksandra",
year = "2020",
abstract = "Vaccines can have heterologous effects on the immune system, i.e., effects other than triggering an immune response against the disease targeted by the vaccine. We investigated whether monoclonal antibodies (mAbs) specific for tetanus could cross-react with Chlamydia and confer heterologous protection against chlamydial infection. The capability of two tetanus-specific mAbs, namely mAb26 and mAb51, to prevent chlamydial infection has been assessed: (i) in vitro, by performing a neutralization assay using human conjunctival epithelial (HCjE) cells infected with Chlamydia trachomatis serovar B, and (ii) in vivo, by using a guinea pig model of Chlamydiacaviae-induced inclusion conjunctivitis. The mAb26 has been superior in comparison with mAb51 in the prevention of chlamydial infection in HCjE cells. The mAb26 has conferred ≈40% inhibition of the infection, compared to less than 5% inhibition in the presence of the mAb51. In vivo, mAb26 significantly diminished ocular pathology intensity in guinea pigs infected with C. caviae compared to either the mAb51-treated or sham-treated guinea pigs. Our data provide insights that tetanus immunization generates antibodies which induce heterologous chlamydial immunity and promote protection beyond the intended target pathogen.",
journal = "Vaccines",
title = "Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia",
volume = "8",
number = "4",
pages = "719",
doi = "10.3390/vaccines8040719"
}

Stojanović, M. M., Lukić, I., Marinković, E., Kovačević, A., Miljković, R., Tobias, J., Schabussova, I., Zlatović, M., Barisani-Asenbauer, T., Wiedermann, U.,& Inic-Kanada, A.. (2020). Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. in Vaccines, 8(4), 719.
https://doi.org/10.3390/vaccines8040719

Stojanović MM, Lukić I, Marinković E, Kovačević A, Miljković R, Tobias J, Schabussova I, Zlatović M, Barisani-Asenbauer T, Wiedermann U, Inic-Kanada A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. in Vaccines. 2020;8(4):719.
doi:10.3390/vaccines8040719 .

Stojanović, Marijana M., Lukić, Ivana, Marinković, Emilija, Kovačević, Ana, Miljković, Radmila, Tobias, Joshua, Schabussova, Irma, Zlatović, Mario, Barisani-Asenbauer, Talin, Wiedermann, Ursula, Inic-Kanada, Aleksandra, "Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia" in Vaccines, 8, no. 4 (2020):719,
https://doi.org/10.3390/vaccines8040719 . .