TY  - DATA
AU  - Gligorijević, Nikola
AU  - Lujić, Tamara
AU  - Mutić, Tamara
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - de Guzman, Maria Krishna
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2024
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6469
AB  - Research data for the unpublished paper: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. Densitometry analysis of ovalbumin band intensity after digestion in the presence of microplastics on SDS-PAGE gels.
T1  - Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6469
ER  - 

@misc{
author = "Gligorijević, Nikola and Lujić, Tamara and Mutić, Tamara and Vasović, Tamara and Aćimović, Jelena and de Guzman, Maria Krishna and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2024",
abstract = "Research data for the unpublished paper: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. Densitometry analysis of ovalbumin band intensity after digestion in the presence of microplastics on SDS-PAGE gels.",
title = "Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6469"
}

Gligorijević, N., Lujić, T., Mutić, T., Vasović, T., Aćimović, J., de Guzman, M. K., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2024). Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. .
https://hdl.handle.net/21.15107/rcub_cherry_6469

Gligorijević N, Lujić T, Mutić T, Vasović T, Aćimović J, de Guzman MK, Stanić-Vučinić D, Ćirković-Veličković T. Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. 2024;.
https://hdl.handle.net/21.15107/rcub_cherry_6469 .

Gligorijević, Nikola, Lujić, Tamara, Mutić, Tamara, Vasović, Tamara, Aćimović, Jelena, de Guzman, Maria Krishna, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties" (2024),
https://hdl.handle.net/21.15107/rcub_cherry_6469 .

TY  - DATA
AU  - Gligorijević, Nikola
AU  - Lujić, Tamara
AU  - Mutić, Tamara
AU  - Vasović, Tamara
AU  - de Guzman, Maria Krishna
AU  - Aćimović, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2024
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6465
AB  - Data used for the analysis of pepsin interaction with polystyrene microplastic of 10 and 100 µm in size. Binding isotherms data were used for the construction of binding isotherms from which binding between pepsin and polystyrene microplastic was described. Structural features of pepsin in the presence of polystyrene were analysed by CD spectrometry and spectrofluorimetry.
T1  - Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6465
ER  - 

@misc{
author = "Gligorijević, Nikola and Lujić, Tamara and Mutić, Tamara and Vasović, Tamara and de Guzman, Maria Krishna and Aćimović, Jelena and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2024",
abstract = "Data used for the analysis of pepsin interaction with polystyrene microplastic of 10 and 100 µm in size. Binding isotherms data were used for the construction of binding isotherms from which binding between pepsin and polystyrene microplastic was described. Structural features of pepsin in the presence of polystyrene were analysed by CD spectrometry and spectrofluorimetry.",
title = "Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6465"
}

Gligorijević, N., Lujić, T., Mutić, T., Vasović, T., de Guzman, M. K., Aćimović, J., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2024). Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. .
https://hdl.handle.net/21.15107/rcub_cherry_6465

Gligorijević N, Lujić T, Mutić T, Vasović T, de Guzman MK, Aćimović J, Stanić-Vučinić D, Ćirković-Veličković T. Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. 2024;.
https://hdl.handle.net/21.15107/rcub_cherry_6465 .

Gligorijević, Nikola, Lujić, Tamara, Mutić, Tamara, Vasović, Tamara, de Guzman, Maria Krishna, Aćimović, Jelena, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties" (2024),
https://hdl.handle.net/21.15107/rcub_cherry_6465 .

TY  - JOUR
AU  - Khulal, Urmila
AU  - Đukić, Teodora
AU  - Smiljanić, Katarina
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Rajković, Andreja
AU  - Ćirković-Veličković, Tanja
PY  - 2024
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6437
AB  - Meat samples were subjected to thermal processing combined with simulated INFOGEST in vitro gastrointestinal (GI) digestion. Protein modifications (PMs) were screened with commercially available PM-specific antibodies. Specific proteins at 20, 37, 50, and 65 kDa react to more than 3 PM-specific antibodies among meat proteins. Lysine methylation and methionine oxidation were the most prominent PMs in WB. Mass spectrometry confirmed bands at ≈20 kDa as allergenic proteins: sarcoplasmic calcium-binding protein in oyster, 37 kDa as tropomyosin in shrimp, oyster, and abalone. MS-identified PMs of shellfish allergens were aligned to the IgE binding epitopes. GI digestion-resistant peptides of shellfish proteins were identified as paramyosins in oyster and abalone and SBP in shrimp. Our results point to the high susceptibility of immunodominant epitopes of major shellfish allergens to PMs. In TPM, saturation of oxidative modification increases with thermal processing resulting in higher susceptibility of TPM to gastric digestion.
PB  - Elsevier
T2  - Journal of Functional Foods
T1  - Protein modifications screening of raw and thermally treated meat gastrointestinal digesta
VL  - 113
SP  - 106052
DO  - 10.1016/j.jff.2024.106052
ER  - 

@article{
author = "Khulal, Urmila and Đukić, Teodora and Smiljanić, Katarina and Vasović, Tamara and Aćimović, Jelena and Rajković, Andreja and Ćirković-Veličković, Tanja",
year = "2024",
abstract = "Meat samples were subjected to thermal processing combined with simulated INFOGEST in vitro gastrointestinal (GI) digestion. Protein modifications (PMs) were screened with commercially available PM-specific antibodies. Specific proteins at 20, 37, 50, and 65 kDa react to more than 3 PM-specific antibodies among meat proteins. Lysine methylation and methionine oxidation were the most prominent PMs in WB. Mass spectrometry confirmed bands at ≈20 kDa as allergenic proteins: sarcoplasmic calcium-binding protein in oyster, 37 kDa as tropomyosin in shrimp, oyster, and abalone. MS-identified PMs of shellfish allergens were aligned to the IgE binding epitopes. GI digestion-resistant peptides of shellfish proteins were identified as paramyosins in oyster and abalone and SBP in shrimp. Our results point to the high susceptibility of immunodominant epitopes of major shellfish allergens to PMs. In TPM, saturation of oxidative modification increases with thermal processing resulting in higher susceptibility of TPM to gastric digestion.",
publisher = "Elsevier",
journal = "Journal of Functional Foods",
title = "Protein modifications screening of raw and thermally treated meat gastrointestinal digesta",
volume = "113",
pages = "106052",
doi = "10.1016/j.jff.2024.106052"
}

Khulal, U., Đukić, T., Smiljanić, K., Vasović, T., Aćimović, J., Rajković, A.,& Ćirković-Veličković, T.. (2024). Protein modifications screening of raw and thermally treated meat gastrointestinal digesta. in Journal of Functional Foods
Elsevier., 113, 106052.
https://doi.org/10.1016/j.jff.2024.106052

Khulal U, Đukić T, Smiljanić K, Vasović T, Aćimović J, Rajković A, Ćirković-Veličković T. Protein modifications screening of raw and thermally treated meat gastrointestinal digesta. in Journal of Functional Foods. 2024;113:106052.
doi:10.1016/j.jff.2024.106052 .

Khulal, Urmila, Đukić, Teodora, Smiljanić, Katarina, Vasović, Tamara, Aćimović, Jelena, Rajković, Andreja, Ćirković-Veličković, Tanja, "Protein modifications screening of raw and thermally treated meat gastrointestinal digesta" in Journal of Functional Foods, 113 (2024):106052,
https://doi.org/10.1016/j.jff.2024.106052 . .

TY  - CONF
AU  - Lujić, Tamara
AU  - Gligorijević, Nikola
AU  - Jovanović, Vesna
AU  - Aćimović, Jelena
AU  - Mitić, Dragana
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6424
AB  - Microplastics is abundant in the environment, food and beverages and get ingested by humans. Its complex interplay with proteins lead to formation of corona. Tightly bound proteins represent hard corona, while weaker binding partners are found in soft corona. Separation of hard and soft corona of allergenic proteins of shrimps, eggs and cow’s milk, tropomyosin (TPM), ovalbumin (OVA) and beta-lactoglobulin (BLG) and identification of binding partners by proteomics was aim of our study.
Allergenic proteins were purified from egg white, shrimps and cow’s milk. Binding to polyethylene terephthalate microplastics (PET) (70-100 m) was probed at pH 7 for purified allergens and egg white proteins. After establishment of binding equilibrium, soft and hard corona were separated and analyzed by SDS PAGE, followed by identification of bound proteins by nanoLC-HRMS. Binding of all allergenic proteins was observed in both soft and hard corona. Soft corona contains exclusively intact, full length OVA, TPM and BLG. Hard corona is enriched for truncated OVA and oligomers of TPM. OVA fragments are partially or fully enfolded and have higher level of exposed hydrophobic patches resulting in higher affinity for PET microplastics. In comparison to OVA and TPM, hard corona of BLG is less abundant under similar conditions. BLG is compact globular protein with lower level of exposed hydrophobic patches in comparison to ovalbumin and tropomyosin. In hard corona, trace amounts of contaminating alfa-lactalbumin become enriched. In the presence of egg white protein extract OVA forms both SC and HC on microplastics, being the dominant protein of hard corona (with ovotransferrin). Lysozyme and ovomucin are present only in hard corona. Both proteins are known for their strong bioactivity and represent a small fraction of total egg white proteins.
Our results show that allergenic proteins form hard corona on PET microplastics. Among egg white proteins, minor proteins such as lysozyme and ovomucin become enriched. Denaturing effect of strong binding to microplastics may change functional characteristics of allergens and bioactive proteins of foods and should be further investigated in functional assays.
Acknowledgment: This study was supported by IMPTOX European Union's Horizon 2020 research and innovation program (grant number 965173).
PB  - Italian Proteomics Association
C3  - ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy
T1  - Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics
SP  - 11
EP  - 11
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6424
ER  - 

@conference{
author = "Lujić, Tamara and Gligorijević, Nikola and Jovanović, Vesna and Aćimović, Jelena and Mitić, Dragana and Vasović, Tamara and Stojadinović, Marija and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Microplastics is abundant in the environment, food and beverages and get ingested by humans. Its complex interplay with proteins lead to formation of corona. Tightly bound proteins represent hard corona, while weaker binding partners are found in soft corona. Separation of hard and soft corona of allergenic proteins of shrimps, eggs and cow’s milk, tropomyosin (TPM), ovalbumin (OVA) and beta-lactoglobulin (BLG) and identification of binding partners by proteomics was aim of our study.
Allergenic proteins were purified from egg white, shrimps and cow’s milk. Binding to polyethylene terephthalate microplastics (PET) (70-100 m) was probed at pH 7 for purified allergens and egg white proteins. After establishment of binding equilibrium, soft and hard corona were separated and analyzed by SDS PAGE, followed by identification of bound proteins by nanoLC-HRMS. Binding of all allergenic proteins was observed in both soft and hard corona. Soft corona contains exclusively intact, full length OVA, TPM and BLG. Hard corona is enriched for truncated OVA and oligomers of TPM. OVA fragments are partially or fully enfolded and have higher level of exposed hydrophobic patches resulting in higher affinity for PET microplastics. In comparison to OVA and TPM, hard corona of BLG is less abundant under similar conditions. BLG is compact globular protein with lower level of exposed hydrophobic patches in comparison to ovalbumin and tropomyosin. In hard corona, trace amounts of contaminating alfa-lactalbumin become enriched. In the presence of egg white protein extract OVA forms both SC and HC on microplastics, being the dominant protein of hard corona (with ovotransferrin). Lysozyme and ovomucin are present only in hard corona. Both proteins are known for their strong bioactivity and represent a small fraction of total egg white proteins.
Our results show that allergenic proteins form hard corona on PET microplastics. Among egg white proteins, minor proteins such as lysozyme and ovomucin become enriched. Denaturing effect of strong binding to microplastics may change functional characteristics of allergens and bioactive proteins of foods and should be further investigated in functional assays.
Acknowledgment: This study was supported by IMPTOX European Union's Horizon 2020 research and innovation program (grant number 965173).",
publisher = "Italian Proteomics Association",
journal = "ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy",
title = "Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics",
pages = "11-11",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6424"
}

Lujić, T., Gligorijević, N., Jovanović, V., Aćimović, J., Mitić, D., Vasović, T., Stojadinović, M., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics. in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy
Italian Proteomics Association., 11-11.
https://hdl.handle.net/21.15107/rcub_cherry_6424

Lujić T, Gligorijević N, Jovanović V, Aćimović J, Mitić D, Vasović T, Stojadinović M, Stanić-Vučinić D, Ćirković-Veličković T. Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics. in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy. 2023;:11-11.
https://hdl.handle.net/21.15107/rcub_cherry_6424 .

Lujić, Tamara, Gligorijević, Nikola, Jovanović, Vesna, Aćimović, Jelena, Mitić, Dragana, Vasović, Tamara, Stojadinović, Marija, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics" in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy (2023):11-11,
https://hdl.handle.net/21.15107/rcub_cherry_6424 .

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  - GEN
AU  - Ćirković-Veličković, Tanja
AU  - Gnjatović, Marija
AU  - Ćujić, Danica
AU  - Todorović, Aleksandra
AU  - Stanić-Vučinić, Dragana
AU  - Đukić, Teodora
AU  - Mladenović, Maja
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Krstić-Ristivojević, Maja
AU  - Jovanović, Vesna
AU  - Simović, Ana
AU  - Radosavljević, Jelena
AU  - Aćimović, Jelena M.
AU  - Radomirović, Mirjana Ž.
AU  - Stojanović, Marijana
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6014
AB  - Нови корона вирус (SARS CoV-2) који се појавио у Вухану 2019. године припада групи једноланчаних РНК вируса [1]. Представља нови инфективни агенс за хуману популацију и веома је брзо детектован у великом броју земаља. Узрочник је респираторних инфекција које могу да буду праћене и веома тешком клиничком сликом. Брзо ширење, одсуство имунитета на овај вирус и одсуство поузданих тестова за детекцију вируса у тренутку избијања пандемије су болест изазвану овим вирусом брзо претворили у здравствени и друштвени проблем највишег приоритета на глобалном нивоу. Иако су највеће биотехнолошке компаније убрзано почеле са развојем и масовном производњом дијагностичких тестова и вакцина, њихова доступност у тренуцима највеће потражње је и даље недовољна, а цене истих су лимитирајући фактор за бољу контролу болести и ширења пандемије [2]. Развој сопствених и одржива производња тестова и вакцина за COVID-19 су од великог друштвеног значаја. Важан предуслов за одрживу производњу тестова је доступност рекомбинантних антигена вируса и могућност производње истих на великој скали за потребе производње домаћих тестова. Овим техничким решењем се описује добијање два кључна антигена новог корона вируса рекомбинантном технологијом и њихова примена у серолошком ЕЛИСА тесту који производи Институт за примену нуклеарне енергије, ИНЕП, као и за добијање реагенаса за детекцију антигена новог корона вируса (специфичних антитела). У првој фази, оптимизоване су секвенце протеина које су подигле осетљивост постојећих серолошких тестова. Иновативност нашег приступа се огледа и у разрађеним експерименталним протоколима за добијање рекомбинантних протеина нуклеокапсида на великој скали, као и у солубилној форми, што олакшава поступак пречишћавања. Избор фрагмента нуклеокапсида који се хетеролого експримира у солубилној форми, а специфично детектује антитела и генерише јак имуни одговор током имунизације животиња (имуногеност) на основу прегледа познатих епитопских секвенци је кључна иновација овог техничког решења. Ово је први пример успешно примењеног рекомбинатног протеина произведеног у Србији у дијагностичком тесту који је регистрован
код Агенције за лекове и медицинска средства Републике Србије (број решења 515-02-02370-21-002), а који је примену нашао и на међународном нивоу.
T1  - Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6014
ER  - 

@misc{
author = "Ćirković-Veličković, Tanja and Gnjatović, Marija and Ćujić, Danica and Todorović, Aleksandra and Stanić-Vučinić, Dragana and Đukić, Teodora and Mladenović, Maja and Vasović, Tamara and Stojadinović, Marija and Krstić-Ristivojević, Maja and Jovanović, Vesna and Simović, Ana and Radosavljević, Jelena and Aćimović, Jelena M. and Radomirović, Mirjana Ž. and Stojanović, Marijana",
year = "2023",
abstract = "Нови корона вирус (SARS CoV-2) који се појавио у Вухану 2019. године припада групи једноланчаних РНК вируса [1]. Представља нови инфективни агенс за хуману популацију и веома је брзо детектован у великом броју земаља. Узрочник је респираторних инфекција које могу да буду праћене и веома тешком клиничком сликом. Брзо ширење, одсуство имунитета на овај вирус и одсуство поузданих тестова за детекцију вируса у тренутку избијања пандемије су болест изазвану овим вирусом брзо претворили у здравствени и друштвени проблем највишег приоритета на глобалном нивоу. Иако су највеће биотехнолошке компаније убрзано почеле са развојем и масовном производњом дијагностичких тестова и вакцина, њихова доступност у тренуцима највеће потражње је и даље недовољна, а цене истих су лимитирајући фактор за бољу контролу болести и ширења пандемије [2]. Развој сопствених и одржива производња тестова и вакцина за COVID-19 су од великог друштвеног значаја. Важан предуслов за одрживу производњу тестова је доступност рекомбинантних антигена вируса и могућност производње истих на великој скали за потребе производње домаћих тестова. Овим техничким решењем се описује добијање два кључна антигена новог корона вируса рекомбинантном технологијом и њихова примена у серолошком ЕЛИСА тесту који производи Институт за примену нуклеарне енергије, ИНЕП, као и за добијање реагенаса за детекцију антигена новог корона вируса (специфичних антитела). У првој фази, оптимизоване су секвенце протеина које су подигле осетљивост постојећих серолошких тестова. Иновативност нашег приступа се огледа и у разрађеним експерименталним протоколима за добијање рекомбинантних протеина нуклеокапсида на великој скали, као и у солубилној форми, што олакшава поступак пречишћавања. Избор фрагмента нуклеокапсида који се хетеролого експримира у солубилној форми, а специфично детектује антитела и генерише јак имуни одговор током имунизације животиња (имуногеност) на основу прегледа познатих епитопских секвенци је кључна иновација овог техничког решења. Ово је први пример успешно примењеног рекомбинатног протеина произведеног у Србији у дијагностичком тесту који је регистрован
код Агенције за лекове и медицинска средства Републике Србије (број решења 515-02-02370-21-002), а који је примену нашао и на међународном нивоу.",
title = "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6014"
}

Ćirković-Veličković, T., Gnjatović, M., Ćujić, D., Todorović, A., Stanić-Vučinić, D., Đukić, T., Mladenović, M., Vasović, T., Stojadinović, M., Krstić-Ristivojević, M., Jovanović, V., Simović, A., Radosavljević, J., Aćimović, J. M., Radomirović, M. Ž.,& Stojanović, M.. (2023). Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. .
https://hdl.handle.net/21.15107/rcub_cherry_6014

Ćirković-Veličković T, Gnjatović M, Ćujić D, Todorović A, Stanić-Vučinić D, Đukić T, Mladenović M, Vasović T, Stojadinović M, Krstić-Ristivojević M, Jovanović V, Simović A, Radosavljević J, Aćimović JM, Radomirović MŽ, Stojanović M. Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_6014 .

Ćirković-Veličković, Tanja, Gnjatović, Marija, Ćujić, Danica, Todorović, Aleksandra, Stanić-Vučinić, Dragana, Đukić, Teodora, Mladenović, Maja, Vasović, Tamara, Stojadinović, Marija, Krstić-Ristivojević, Maja, Jovanović, Vesna, Simović, Ana, Radosavljević, Jelena, Aćimović, Jelena M., Radomirović, Mirjana Ž., Stojanović, Marijana, "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus" (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6014 .

TY  - JOUR
AU  - Takić, Marija M.
AU  - Jovanović, Vesna B.
AU  - Pavićević, Ivan D.
AU  - Uzelac, Tamara N.
AU  - Aćimović, Jelena M.
AU  - Ristić-Medić, Danijela
AU  - Mandić, Ljuba M.
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2054
AB  - The interaction of polyphenolic molecules with human serum albumin (HSA) could lead to changes in the reactivity of the HSA Cys34 thiol group (HSA-SH). The influences of enterolactone (EL) and enterodiol (ED) binding on HSA-SH reactivity in fatty acid (FA)-free HSA, and in HSA with bound stearic acid (S) in S/HSA molar ratios of 1 : 1 and 4 : 1, were investigated by the determination of the pseudo first order rate constants (k') for the thiol reaction with 5,5'-dithiobis-(2-nitrobenzoic acid). The binding affinities and binding sites of EL and ED were also determined, using fluorescence measurements of the intrinsic fluorescence of Trp214 and diazepam (binding site marker). EL and ED binding to HSA increased the reactivity of HSA-SH in all assayed HSA-enterolignan complexes by 9.1-33.1%. The strongest effects were obtained for FA-free HSA-enterolignan complexes. S modulated/reduced the effect of EL on HSA-SH reactivity, while its influence on the effect of ED was negligible. The binding of enterolignans to HSA was investigated: the binding constants were the highest for FA-free HSA (EL: 11.64 x 10(4) M-1 and ED: 5.59 x 10(4) M-1 at 37 degrees C) and the lowest for S/HSA 4 : 1-enterolignan complexes (EL: 2.43 x 10(4) M-1 and ED: 1.92 x 10(4) M-1). When the S/HSA ratio was increased, the binding affinities and number of binding sites for EL and ED were decreased. At the same time, a high correlation between binding constants and increased Cys34 reactivity was found (r = 0.974). Competitive experiments using diazepam indicated that the binding of ED and of EL was located in the hydrophobic pocket of site II in HSA. Overall, it is evident that stearic acid could modulate the enterolignan effects on HSA-SH reactivity as well as their binding to HSA. This finding could be important for pharmacokinetics and the expression of enterolignan antioxidant effects in vivo after an intake of lignan rich food.
PB  - Royal Soc Chemistry, Cambridge
T2  - Food and Function
T1  - Binding of enterolactone and enterodiol to human serum albumin: increase of cysteine-34 thiol group reactivity
VL  - 7
IS  - 2
SP  - 1217
EP  - 1226
DO  - 10.1039/c5fo01346a
ER  - 

@article{
author = "Takić, Marija M. and Jovanović, Vesna B. and Pavićević, Ivan D. and Uzelac, Tamara N. and Aćimović, Jelena M. and Ristić-Medić, Danijela and Mandić, Ljuba M.",
year = "2016",
abstract = "The interaction of polyphenolic molecules with human serum albumin (HSA) could lead to changes in the reactivity of the HSA Cys34 thiol group (HSA-SH). The influences of enterolactone (EL) and enterodiol (ED) binding on HSA-SH reactivity in fatty acid (FA)-free HSA, and in HSA with bound stearic acid (S) in S/HSA molar ratios of 1 : 1 and 4 : 1, were investigated by the determination of the pseudo first order rate constants (k') for the thiol reaction with 5,5'-dithiobis-(2-nitrobenzoic acid). The binding affinities and binding sites of EL and ED were also determined, using fluorescence measurements of the intrinsic fluorescence of Trp214 and diazepam (binding site marker). EL and ED binding to HSA increased the reactivity of HSA-SH in all assayed HSA-enterolignan complexes by 9.1-33.1%. The strongest effects were obtained for FA-free HSA-enterolignan complexes. S modulated/reduced the effect of EL on HSA-SH reactivity, while its influence on the effect of ED was negligible. The binding of enterolignans to HSA was investigated: the binding constants were the highest for FA-free HSA (EL: 11.64 x 10(4) M-1 and ED: 5.59 x 10(4) M-1 at 37 degrees C) and the lowest for S/HSA 4 : 1-enterolignan complexes (EL: 2.43 x 10(4) M-1 and ED: 1.92 x 10(4) M-1). When the S/HSA ratio was increased, the binding affinities and number of binding sites for EL and ED were decreased. At the same time, a high correlation between binding constants and increased Cys34 reactivity was found (r = 0.974). Competitive experiments using diazepam indicated that the binding of ED and of EL was located in the hydrophobic pocket of site II in HSA. Overall, it is evident that stearic acid could modulate the enterolignan effects on HSA-SH reactivity as well as their binding to HSA. This finding could be important for pharmacokinetics and the expression of enterolignan antioxidant effects in vivo after an intake of lignan rich food.",
publisher = "Royal Soc Chemistry, Cambridge",
journal = "Food and Function",
title = "Binding of enterolactone and enterodiol to human serum albumin: increase of cysteine-34 thiol group reactivity",
volume = "7",
number = "2",
pages = "1217-1226",
doi = "10.1039/c5fo01346a"
}

Takić, M. M., Jovanović, V. B., Pavićević, I. D., Uzelac, T. N., Aćimović, J. M., Ristić-Medić, D.,& Mandić, L. M.. (2016). Binding of enterolactone and enterodiol to human serum albumin: increase of cysteine-34 thiol group reactivity. in Food and Function
Royal Soc Chemistry, Cambridge., 7(2), 1217-1226.
https://doi.org/10.1039/c5fo01346a

Takić MM, Jovanović VB, Pavićević ID, Uzelac TN, Aćimović JM, Ristić-Medić D, Mandić LM. Binding of enterolactone and enterodiol to human serum albumin: increase of cysteine-34 thiol group reactivity. in Food and Function. 2016;7(2):1217-1226.
doi:10.1039/c5fo01346a .

Takić, Marija M., Jovanović, Vesna B., Pavićević, Ivan D., Uzelac, Tamara N., Aćimović, Jelena M., Ristić-Medić, Danijela, Mandić, Ljuba M., "Binding of enterolactone and enterodiol to human serum albumin: increase of cysteine-34 thiol group reactivity" in Food and Function, 7, no. 2 (2016):1217-1226,
https://doi.org/10.1039/c5fo01346a . .

TY  - CONF
AU  - Aćimović, Jelena M.
AU  - Penezić, Ana Z.
AU  - Pavićević, Ivan D.
AU  - Jovanović, Vesna B.
AU  - Takić, Marija M.
AU  - Uzelac, T. N.
AU  - Mandić, Ljuba M.
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2307
PB  - Wiley-Blackwell, Hoboken
C3  - FEBS Journal / Federation of European of Biochemical Societies
T1  - Binding of FAs and Cu(II) ions to HSA changes its Cys34 thiol group antioxidant capacity and carbonylation pattern with methylglyoxal
VL  - 283
SP  - 417
EP  - 417
UR  - https://hdl.handle.net/21.15107/rcub_cherry_2307
ER  - 

@conference{
author = "Aćimović, Jelena M. and Penezić, Ana Z. and Pavićević, Ivan D. and Jovanović, Vesna B. and Takić, Marija M. and Uzelac, T. N. and Mandić, Ljuba M.",
year = "2016",
publisher = "Wiley-Blackwell, Hoboken",
journal = "FEBS Journal / Federation of European of Biochemical Societies",
title = "Binding of FAs and Cu(II) ions to HSA changes its Cys34 thiol group antioxidant capacity and carbonylation pattern with methylglyoxal",
volume = "283",
pages = "417-417",
url = "https://hdl.handle.net/21.15107/rcub_cherry_2307"
}

Aćimović, J. M., Penezić, A. Z., Pavićević, I. D., Jovanović, V. B., Takić, M. M., Uzelac, T. N.,& Mandić, L. M.. (2016). Binding of FAs and Cu(II) ions to HSA changes its Cys34 thiol group antioxidant capacity and carbonylation pattern with methylglyoxal. in FEBS Journal / Federation of European of Biochemical Societies
Wiley-Blackwell, Hoboken., 283, 417-417.
https://hdl.handle.net/21.15107/rcub_cherry_2307

Aćimović JM, Penezić AZ, Pavićević ID, Jovanović VB, Takić MM, Uzelac TN, Mandić LM. Binding of FAs and Cu(II) ions to HSA changes its Cys34 thiol group antioxidant capacity and carbonylation pattern with methylglyoxal. in FEBS Journal / Federation of European of Biochemical Societies. 2016;283:417-417.
https://hdl.handle.net/21.15107/rcub_cherry_2307 .

Aćimović, Jelena M., Penezić, Ana Z., Pavićević, Ivan D., Jovanović, Vesna B., Takić, Marija M., Uzelac, T. N., Mandić, Ljuba M., "Binding of FAs and Cu(II) ions to HSA changes its Cys34 thiol group antioxidant capacity and carbonylation pattern with methylglyoxal" in FEBS Journal / Federation of European of Biochemical Societies, 283 (2016):417-417,
https://hdl.handle.net/21.15107/rcub_cherry_2307 .

TY  - JOUR
AU  - Pavićević, Ivan D.
AU  - Jovanović, Vesna B.
AU  - Takić, Marija M.
AU  - Aćimović, Jelena M.
AU  - Penezić, Ana Z.
AU  - Mandić, Ljuba M.
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2399
AB  - Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states. (C) 2016 Elsevier B.V. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Pharmaceutical and Biomedical Analysis
T1  - Quantification of total content of non-esterified fatty acids bound to human serum albumin
VL  - 129
SP  - 43
EP  - 49
DO  - 10.1016/j.jpba.2016.06.043
ER  - 

@article{
author = "Pavićević, Ivan D. and Jovanović, Vesna B. and Takić, Marija M. and Aćimović, Jelena M. and Penezić, Ana Z. and Mandić, Ljuba M.",
year = "2016",
abstract = "Non-esterified fatty acids bound to the human serum albumin (HSA) contribute to several HSAs properties of special concern in pathologies, for instance to the reactivity of the free HSA-Cys34 thiol group (important antioxidative thiol pool in plasma), and to the affinity for binding of molecules and ions (for example cobalt as a prominent biomarker in heart ischemia). Therefore, the method for determination of FAs bound to HSA was developed. FAs were released from HSA (previously isolated from serum by ammonium sulfate precipitation) using acidic copper(II) sulfate in phosphoric acid, extracted by n-heptane-chloroform (4:1, v/v) mixture, spotted on TL silica-gel and then developed with n-heptane-chloroform-acetic acid (5:3:03, v/v/v). Common office flatbed scanner and software solution for densitometric image analysis, developed in R, were used. The linearity of calibration curve in concentration range from 0.1 to 5.0 mmol/L stearic acid was achieved. The method was proved to be precise (with RSD of 1.4-4.7%) and accurate. Accuracy was examined by standard addition method (recoveries 97.2-102.5%) and by comparison to results of GC. The method is sample saving, technically less demanding, and cheap, and therefore suitable for determination of FAs/HSA ratio when elevated concentrations of free FAs are reliable diagnostic/risk parameter of pathological states. (C) 2016 Elsevier B.V. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Pharmaceutical and Biomedical Analysis",
title = "Quantification of total content of non-esterified fatty acids bound to human serum albumin",
volume = "129",
pages = "43-49",
doi = "10.1016/j.jpba.2016.06.043"
}

Pavićević, I. D., Jovanović, V. B., Takić, M. M., Aćimović, J. M., Penezić, A. Z.,& Mandić, L. M.. (2016). Quantification of total content of non-esterified fatty acids bound to human serum albumin. in Journal of Pharmaceutical and Biomedical Analysis
Elsevier Science Bv, Amsterdam., 129, 43-49.
https://doi.org/10.1016/j.jpba.2016.06.043

Pavićević ID, Jovanović VB, Takić MM, Aćimović JM, Penezić AZ, Mandić LM. Quantification of total content of non-esterified fatty acids bound to human serum albumin. in Journal of Pharmaceutical and Biomedical Analysis. 2016;129:43-49.
doi:10.1016/j.jpba.2016.06.043 .

Pavićević, Ivan D., Jovanović, Vesna B., Takić, Marija M., Aćimović, Jelena M., Penezić, Ana Z., Mandić, Ljuba M., "Quantification of total content of non-esterified fatty acids bound to human serum albumin" in Journal of Pharmaceutical and Biomedical Analysis, 129 (2016):43-49,
https://doi.org/10.1016/j.jpba.2016.06.043 . .

TY  - JOUR
AU  - Penezić, Ana Z.
AU  - Jovanović, Vesna B.
AU  - Pavićević, Ivan D.
AU  - Aćimović, Jelena M.
AU  - Mandić, Ljuba M.
PY  - 2015
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1979
AB  - The potential of carbonylation with methylglyoxal to alter HSA's binding affinity for copper(II) ions and its influence on the release of copper(II) ions from copper-HSA complexes were studied. The affinity of HSA to coordinate copper(II) decreased upon carbonylation of the Cys34-SH group. Carbonylation of copper-HSA complexes caused a decrease in Cys34-SH content, conformational changes and the release of copper(II) ions. The ratio between the percentage of reduction in the Cys34-SH group content and the percentage of release of copper(II) from complexes is 2.12 +/- 0.28. Because the same ratio (1.96 +/- 0.36) was obtained upon oxidation of the Cys34-SH group (with no changes in HSA conformation), the binding/release of copper (II) by HSA depended mainly on the redox state of the Cys34-SH group. The contents of Cys34-SH and HSA-bound copper(II) ions in the diabetic group (0.457 +/- 0.081 mol SH per mol HSA, 10.7 +/- 0.01 mmol per mol HSA, resp.) were significantly lower (p  lt  0.01) compared to the control group (0.609 +/- 0.027 mol SH per mol HSA; 13.4 +/- 0.01 mmol per mol HSA, resp.). Very strong correlations between the values for HSA-SH and glycated haemoglobin, HbA1c, (R = -0.803, p  lt  0.01), and between the values for the HSA-bound copper(II) content and HSA-SH content (R = 0.841, p  lt  0.002) were found in the diabetic group. Thus, HSA carbonylation leads to decrease in HSA-SH content and to the impairment of its copper(II) binding capacity that could contribute to further enhancement of oxidative and carbonyl stress in diabetes (as well as in other diseases with carbonyl stress).
PB  - Royal Soc Chemistry, Cambridge
T2  - Metallomics
T1  - HSA carbonylation with methylglyoxal and the binding/release of copper(II) ions
VL  - 7
IS  - 10
SP  - 1431
EP  - 1438
DO  - 10.1039/c5mt00159e
ER  - 

@article{
author = "Penezić, Ana Z. and Jovanović, Vesna B. and Pavićević, Ivan D. and Aćimović, Jelena M. and Mandić, Ljuba M.",
year = "2015",
abstract = "The potential of carbonylation with methylglyoxal to alter HSA's binding affinity for copper(II) ions and its influence on the release of copper(II) ions from copper-HSA complexes were studied. The affinity of HSA to coordinate copper(II) decreased upon carbonylation of the Cys34-SH group. Carbonylation of copper-HSA complexes caused a decrease in Cys34-SH content, conformational changes and the release of copper(II) ions. The ratio between the percentage of reduction in the Cys34-SH group content and the percentage of release of copper(II) from complexes is 2.12 +/- 0.28. Because the same ratio (1.96 +/- 0.36) was obtained upon oxidation of the Cys34-SH group (with no changes in HSA conformation), the binding/release of copper (II) by HSA depended mainly on the redox state of the Cys34-SH group. The contents of Cys34-SH and HSA-bound copper(II) ions in the diabetic group (0.457 +/- 0.081 mol SH per mol HSA, 10.7 +/- 0.01 mmol per mol HSA, resp.) were significantly lower (p  lt  0.01) compared to the control group (0.609 +/- 0.027 mol SH per mol HSA; 13.4 +/- 0.01 mmol per mol HSA, resp.). Very strong correlations between the values for HSA-SH and glycated haemoglobin, HbA1c, (R = -0.803, p  lt  0.01), and between the values for the HSA-bound copper(II) content and HSA-SH content (R = 0.841, p  lt  0.002) were found in the diabetic group. Thus, HSA carbonylation leads to decrease in HSA-SH content and to the impairment of its copper(II) binding capacity that could contribute to further enhancement of oxidative and carbonyl stress in diabetes (as well as in other diseases with carbonyl stress).",
publisher = "Royal Soc Chemistry, Cambridge",
journal = "Metallomics",
title = "HSA carbonylation with methylglyoxal and the binding/release of copper(II) ions",
volume = "7",
number = "10",
pages = "1431-1438",
doi = "10.1039/c5mt00159e"
}

Penezić, A. Z., Jovanović, V. B., Pavićević, I. D., Aćimović, J. M.,& Mandić, L. M.. (2015). HSA carbonylation with methylglyoxal and the binding/release of copper(II) ions. in Metallomics
Royal Soc Chemistry, Cambridge., 7(10), 1431-1438.
https://doi.org/10.1039/c5mt00159e

Penezić AZ, Jovanović VB, Pavićević ID, Aćimović JM, Mandić LM. HSA carbonylation with methylglyoxal and the binding/release of copper(II) ions. in Metallomics. 2015;7(10):1431-1438.
doi:10.1039/c5mt00159e .

Penezić, Ana Z., Jovanović, Vesna B., Pavićević, Ivan D., Aćimović, Jelena M., Mandić, Ljuba M., "HSA carbonylation with methylglyoxal and the binding/release of copper(II) ions" in Metallomics, 7, no. 10 (2015):1431-1438,
https://doi.org/10.1039/c5mt00159e . .

TY  - JOUR
AU  - Pavićević, Ivan D.
AU  - Jovanović, Vesna B.
AU  - Takić, Marija M.
AU  - Penezić, Ana Z.
AU  - Aćimović, Jelena M.
AU  - Mandić, Ljuba M.
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1895
AB  - Fatty acids (FAs) binding to human serum albumin (HSA) could lead to the changes of Cys-34 thiol group accessibility and reactivity, i.e. its scavenger capacity and antioxidant property. The influence of saturated, mono and poly unsaturated, and fish oil FAs binding to HSA on the carbonylation level and the reactivity of HSA-SH and HSA modified with methylglyoxal (MG-HSA-SH) was investigated. Changes of thiol group reactivity were followed by determination of pseudo first order rate constant (k') for thiols reaction with 5,5'-dithiobis(2-nitrobenzoic acid). HSA changes were monitored using native PAG electrophoresis and fluorescence spectroscopy. For FA/HSA molar ratios screening, qTLC and GC were used. FAs increase thiol group carbonylation levels from 8% to 20%. The k' values obtained for FAs-free HSA-SH and FAs-free MG-HSA-SH are almost equal (7.5 x 10(-3) and 7.7 x 10(-3) resp.). Binding of all FAs amplify the reactivity (k' values from 14.6 x 10(-3) to 26.0 x 10(-3) s(-1)) of HSA-SH group for 2-3.5 times in the order: palmitic, docosahexaenoic, fish oil extract, stearic, oleic, myristic and eicosapentaenoic acid, due to HSA conformational changes. FAs-bound MG-HSA-SH samples follow that pattern, but their k' values (from 9.8 x 10(-3) to 14.3 x 10(-3) s(-1)) were lower compared to unmodified HSA due to additional conformation changes of HSA molecules during carbonylation. Carbonylation level and reactivity of Cys34 thiol group of unmodified and carbonylated HSA depend on type of FAs bound to HSA, which implies the possibility for modulation of -SH reactivity (scavenger capacity and antioxidant property) by FAs as a supplement. (C) 2014 Elsevier Ireland Ltd. All rights reserved.
PB  - Elsevier Ireland Ltd, Clare
T2  - Chemico-biological Interactions
T1  - Fatty acids binding to human serum albumin: Changes of reactivity and glycation level of Cysteine-34 free thiol group with methylglyoxal
VL  - 224
SP  - 42
EP  - 50
DO  - 10.1016/j.cbi.2014.10.008
ER  - 

@article{
author = "Pavićević, Ivan D. and Jovanović, Vesna B. and Takić, Marija M. and Penezić, Ana Z. and Aćimović, Jelena M. and Mandić, Ljuba M.",
year = "2014",
abstract = "Fatty acids (FAs) binding to human serum albumin (HSA) could lead to the changes of Cys-34 thiol group accessibility and reactivity, i.e. its scavenger capacity and antioxidant property. The influence of saturated, mono and poly unsaturated, and fish oil FAs binding to HSA on the carbonylation level and the reactivity of HSA-SH and HSA modified with methylglyoxal (MG-HSA-SH) was investigated. Changes of thiol group reactivity were followed by determination of pseudo first order rate constant (k') for thiols reaction with 5,5'-dithiobis(2-nitrobenzoic acid). HSA changes were monitored using native PAG electrophoresis and fluorescence spectroscopy. For FA/HSA molar ratios screening, qTLC and GC were used. FAs increase thiol group carbonylation levels from 8% to 20%. The k' values obtained for FAs-free HSA-SH and FAs-free MG-HSA-SH are almost equal (7.5 x 10(-3) and 7.7 x 10(-3) resp.). Binding of all FAs amplify the reactivity (k' values from 14.6 x 10(-3) to 26.0 x 10(-3) s(-1)) of HSA-SH group for 2-3.5 times in the order: palmitic, docosahexaenoic, fish oil extract, stearic, oleic, myristic and eicosapentaenoic acid, due to HSA conformational changes. FAs-bound MG-HSA-SH samples follow that pattern, but their k' values (from 9.8 x 10(-3) to 14.3 x 10(-3) s(-1)) were lower compared to unmodified HSA due to additional conformation changes of HSA molecules during carbonylation. Carbonylation level and reactivity of Cys34 thiol group of unmodified and carbonylated HSA depend on type of FAs bound to HSA, which implies the possibility for modulation of -SH reactivity (scavenger capacity and antioxidant property) by FAs as a supplement. (C) 2014 Elsevier Ireland Ltd. All rights reserved.",
publisher = "Elsevier Ireland Ltd, Clare",
journal = "Chemico-biological Interactions",
title = "Fatty acids binding to human serum albumin: Changes of reactivity and glycation level of Cysteine-34 free thiol group with methylglyoxal",
volume = "224",
pages = "42-50",
doi = "10.1016/j.cbi.2014.10.008"
}

Pavićević, I. D., Jovanović, V. B., Takić, M. M., Penezić, A. Z., Aćimović, J. M.,& Mandić, L. M.. (2014). Fatty acids binding to human serum albumin: Changes of reactivity and glycation level of Cysteine-34 free thiol group with methylglyoxal. in Chemico-biological Interactions
Elsevier Ireland Ltd, Clare., 224, 42-50.
https://doi.org/10.1016/j.cbi.2014.10.008

Pavićević ID, Jovanović VB, Takić MM, Penezić AZ, Aćimović JM, Mandić LM. Fatty acids binding to human serum albumin: Changes of reactivity and glycation level of Cysteine-34 free thiol group with methylglyoxal. in Chemico-biological Interactions. 2014;224:42-50.
doi:10.1016/j.cbi.2014.10.008 .

Pavićević, Ivan D., Jovanović, Vesna B., Takić, Marija M., Penezić, Ana Z., Aćimović, Jelena M., Mandić, Ljuba M., "Fatty acids binding to human serum albumin: Changes of reactivity and glycation level of Cysteine-34 free thiol group with methylglyoxal" in Chemico-biological Interactions, 224 (2014):42-50,
https://doi.org/10.1016/j.cbi.2014.10.008 . .

TY  - JOUR
AU  - Jovanović, Vesna B.
AU  - Pavićević, Ivan D.
AU  - Takić, Marija M.
AU  - Penezić-Romanjuk, Ana Z.
AU  - Aćimović, Jelena M.
AU  - Mandić, Ljuba M.
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1476
AB  - During investigation of the changes of the Cys34 thiol group of human serum albumin (HSA) (isolated by affinity chromatography with Cibacron Blue (CB)) in diabetes, we found that the HSA-SH content was higher (11-33%) than the total serum thiol content. The influence of fatty acids (FA) binding to HSA on this discrepancy was investigated in vitro (using fluorescence and CD spectroscopy and GC) and with HSA samples from diabetic (n=20) and control groups (n=17). HSA-bound FA determine the selection of HSA molecules by CB and enhance reactivity and/or accessibility of the SH group. A high content of polyunsaturated FA (35.6%) leads to weaker binding of HSA molecules to CB. Rate constants of DTNB reaction with the SH group of HSA applied to a CB column, bound-HSA and unbound-HSA fractions, were 4.8 x 10(-3), 21.6 x 10(-3), and 11.2 x 10(-3) s(-1), respectively. The HSA-SH group of diabetics is more reactive compared with control individuals (rate constants 20.9 x 10(-3)+/- 4.4 x 10(-3) vs 12.9 x 10(-3)+/- 2.6 x 10(-3) s(-1), P lt 0.05). Recovery values of the SH group obtained after chromatography of HSA with bound stearic acid ranged from 110 to 140%, while those for defatted HSA were from 98.5 to 101.7%. Thus, HSA-bound FA leads to an increase of HSA-SH content and a contribution to total serum thiols, which make the determination of the thiol group unreliable. (C) 2013 Elsevier Inc. All rights reserved.
PB  - Academic Press Inc Elsevier Science, San Diego
T2  - Analytical Biochemistry
T1  - The influence of fatty acids on determination of human serum albumin thiol group
VL  - 448
SP  - 50
EP  - 57
DO  - 10.1016/j.ab.2013.11.030
ER  - 

@article{
author = "Jovanović, Vesna B. and Pavićević, Ivan D. and Takić, Marija M. and Penezić-Romanjuk, Ana Z. and Aćimović, Jelena M. and Mandić, Ljuba M.",
year = "2014",
abstract = "During investigation of the changes of the Cys34 thiol group of human serum albumin (HSA) (isolated by affinity chromatography with Cibacron Blue (CB)) in diabetes, we found that the HSA-SH content was higher (11-33%) than the total serum thiol content. The influence of fatty acids (FA) binding to HSA on this discrepancy was investigated in vitro (using fluorescence and CD spectroscopy and GC) and with HSA samples from diabetic (n=20) and control groups (n=17). HSA-bound FA determine the selection of HSA molecules by CB and enhance reactivity and/or accessibility of the SH group. A high content of polyunsaturated FA (35.6%) leads to weaker binding of HSA molecules to CB. Rate constants of DTNB reaction with the SH group of HSA applied to a CB column, bound-HSA and unbound-HSA fractions, were 4.8 x 10(-3), 21.6 x 10(-3), and 11.2 x 10(-3) s(-1), respectively. The HSA-SH group of diabetics is more reactive compared with control individuals (rate constants 20.9 x 10(-3)+/- 4.4 x 10(-3) vs 12.9 x 10(-3)+/- 2.6 x 10(-3) s(-1), P lt 0.05). Recovery values of the SH group obtained after chromatography of HSA with bound stearic acid ranged from 110 to 140%, while those for defatted HSA were from 98.5 to 101.7%. Thus, HSA-bound FA leads to an increase of HSA-SH content and a contribution to total serum thiols, which make the determination of the thiol group unreliable. (C) 2013 Elsevier Inc. All rights reserved.",
publisher = "Academic Press Inc Elsevier Science, San Diego",
journal = "Analytical Biochemistry",
title = "The influence of fatty acids on determination of human serum albumin thiol group",
volume = "448",
pages = "50-57",
doi = "10.1016/j.ab.2013.11.030"
}

Jovanović, V. B., Pavićević, I. D., Takić, M. M., Penezić-Romanjuk, A. Z., Aćimović, J. M.,& Mandić, L. M.. (2014). The influence of fatty acids on determination of human serum albumin thiol group. in Analytical Biochemistry
Academic Press Inc Elsevier Science, San Diego., 448, 50-57.
https://doi.org/10.1016/j.ab.2013.11.030

Jovanović VB, Pavićević ID, Takić MM, Penezić-Romanjuk AZ, Aćimović JM, Mandić LM. The influence of fatty acids on determination of human serum albumin thiol group. in Analytical Biochemistry. 2014;448:50-57.
doi:10.1016/j.ab.2013.11.030 .

Jovanović, Vesna B., Pavićević, Ivan D., Takić, Marija M., Penezić-Romanjuk, Ana Z., Aćimović, Jelena M., Mandić, Ljuba M., "The influence of fatty acids on determination of human serum albumin thiol group" in Analytical Biochemistry, 448 (2014):50-57,
https://doi.org/10.1016/j.ab.2013.11.030 . .

TY  - JOUR
AU  - Jovanović, Vesna B.
AU  - Aćimović, Jelena M.
AU  - Sreckovic, Vesna S. Dimitrijevic
AU  - Mandić, Ljuba M.
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1647
AB  - It was verified that the serum N-acetyl-beta-D-glucosaminidase (NAG) activity is elevated in diabetes, but there are no reports about changes in the sialic acid (SA) content in the carbohydrate parts of the NAG A form and its influence on the total changes in NAG activity in type 1 diabetes mellitus patients with and without secondary complications. The NAG A form was isolated from the serum of 81 insulin-dependent diabetes mellitus (IDDM) patients with and without secondary complications (retinopathy, polyneuropathy and nephropathy) and 25 healthy persons, and purified and characterised. The content of alpha-2,6-bound SA, the isoenzyme patterns of the purified A form, and the total NAG and A form activities were determined. In all diabetic groups, the sialylation levels of the A form were 2-3.5 times lower compared to control, while their acidities (fractions with pI 4.25-5.1) increased, particularly with progression of secondary complications. Total serum NAG activities and percentages of A form were significantly higher (P  lt  0.001) in all diabetic groups and negatively correlated with the alpha-2,6-bound SA content of the A form. In addition, they decreased as secondary diabetic complications became more complex. The observed changes could be the consequence of structural changes in the A form due to significant increases in its acidity, i.e., negative charge, which originated from groups other than SA.
PB  - Serbian Chemical Soc, Belgrade
T2  - Journal of the Serbian Chemical Society
T1  - How the sialylation level of serum N-acetyl-beta-D-glucosaminidase A form in Type 1 diabetes mellitus influences their activity?
VL  - 79
IS  - 12
SP  - 1491
EP  - 1503
DO  - 10.2298/JSC140430076J
ER  - 

@article{
author = "Jovanović, Vesna B. and Aćimović, Jelena M. and Sreckovic, Vesna S. Dimitrijevic and Mandić, Ljuba M.",
year = "2014",
abstract = "It was verified that the serum N-acetyl-beta-D-glucosaminidase (NAG) activity is elevated in diabetes, but there are no reports about changes in the sialic acid (SA) content in the carbohydrate parts of the NAG A form and its influence on the total changes in NAG activity in type 1 diabetes mellitus patients with and without secondary complications. The NAG A form was isolated from the serum of 81 insulin-dependent diabetes mellitus (IDDM) patients with and without secondary complications (retinopathy, polyneuropathy and nephropathy) and 25 healthy persons, and purified and characterised. The content of alpha-2,6-bound SA, the isoenzyme patterns of the purified A form, and the total NAG and A form activities were determined. In all diabetic groups, the sialylation levels of the A form were 2-3.5 times lower compared to control, while their acidities (fractions with pI 4.25-5.1) increased, particularly with progression of secondary complications. Total serum NAG activities and percentages of A form were significantly higher (P  lt  0.001) in all diabetic groups and negatively correlated with the alpha-2,6-bound SA content of the A form. In addition, they decreased as secondary diabetic complications became more complex. The observed changes could be the consequence of structural changes in the A form due to significant increases in its acidity, i.e., negative charge, which originated from groups other than SA.",
publisher = "Serbian Chemical Soc, Belgrade",
journal = "Journal of the Serbian Chemical Society",
title = "How the sialylation level of serum N-acetyl-beta-D-glucosaminidase A form in Type 1 diabetes mellitus influences their activity?",
volume = "79",
number = "12",
pages = "1491-1503",
doi = "10.2298/JSC140430076J"
}

Jovanović, V. B., Aćimović, J. M., Sreckovic, V. S. D.,& Mandić, L. M.. (2014). How the sialylation level of serum N-acetyl-beta-D-glucosaminidase A form in Type 1 diabetes mellitus influences their activity?. in Journal of the Serbian Chemical Society
Serbian Chemical Soc, Belgrade., 79(12), 1491-1503.
https://doi.org/10.2298/JSC140430076J

Jovanović VB, Aćimović JM, Sreckovic VSD, Mandić LM. How the sialylation level of serum N-acetyl-beta-D-glucosaminidase A form in Type 1 diabetes mellitus influences their activity?. in Journal of the Serbian Chemical Society. 2014;79(12):1491-1503.
doi:10.2298/JSC140430076J .

Jovanović, Vesna B., Aćimović, Jelena M., Sreckovic, Vesna S. Dimitrijevic, Mandić, Ljuba M., "How the sialylation level of serum N-acetyl-beta-D-glucosaminidase A form in Type 1 diabetes mellitus influences their activity?" in Journal of the Serbian Chemical Society, 79, no. 12 (2014):1491-1503,
https://doi.org/10.2298/JSC140430076J . .

TY  - JOUR
AU  - Aćimović, Jelena M.
AU  - Penezić, Ana Z.
AU  - Pavićević, Ivan D.
AU  - Jovanović, Vesna B.
AU  - Mandić, Ljuba M.
PY  - 2014
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1814
AB  - alpha-Oxoaldehydes, which are produced in higher quantities in diabetes, uremia, oxidative stress, inflammation and aging, react with the amino, guanidine and thiol groups of proteins and cause the formation of advanced glycated end-products and protein cross-linking. To prevent these reactions, the efficiency of tow molecular mass thiols with an alpha-amino-beta-mercapto-ethane group (Cys, penicillamine and N-acetylcysteine (NAcCys, with a blocked amino group)) as scavengers of methylglyoxal, compared with glutathione (GSH) and the biguanidine derivative metformin, was investigated. The time courses of the reactions of the aforementioned compounds with methylglyoxal were assayed. The reactivity of their thiol and amino groups decreased in the order of Cys  gt  penicillamine  gt  GSH  gt  NAcCys and penicillamine  gt  Cys  gt  GSH, respectively. Human serum albumin (HSA) carbonylation in the absence or presence of methylglyoxal scavengers were monitored by the determination of the amino, guanidine and thiol groups' contents, as well as by spectrofluorimetry, CD and native and SDS PAGE. Cys and penicillamine were highly efficient in the prevention of the carbonylation of the HSA-amino (for 80%) and guanidine (for 84% and 55%, respectively) groups and the formation of fluorescent AGEs. GSH and metformin exhibited medium efficiency (reduction of amino group's carbonylation for 60% and guanidine for about 30%); the least efficient was NAcCys. The presence of Cys, penicillamine and NAcCys led to an almost complete protection of the HSA-thiol group's carbonylation, whereas metformin was inefficient. The efficiency in the prevention of protein cross-linking increased in the order of metformin, NAcCys  lt  GSH  lt  penicillamine  lt  Cys. Thus, the substances with an alpha-amino-beta-mercapto-ethane group as a pharmacophore exhibit great potential as an efficient methylglyoxal scavengers, and are thus promising compounds for medicinal chemistry. In addition, they protect the HSA-SH group and preserve its antioxidative potential, which is very important for the HSA's function in vivo.
PB  - Royal Soc Chemistry, Cambridge
T2  - Molecular BioSystems
T1  - The efficiency of compounds with alpha-amino-beta-mercapto-ethane group in protection of human serum albumin carbonylation and cross-linking with methylglyoxal
VL  - 10
IS  - 8
SP  - 2166
EP  - 2175
DO  - 10.1039/c4mb00217b
ER  - 

@article{
author = "Aćimović, Jelena M. and Penezić, Ana Z. and Pavićević, Ivan D. and Jovanović, Vesna B. and Mandić, Ljuba M.",
year = "2014",
abstract = "alpha-Oxoaldehydes, which are produced in higher quantities in diabetes, uremia, oxidative stress, inflammation and aging, react with the amino, guanidine and thiol groups of proteins and cause the formation of advanced glycated end-products and protein cross-linking. To prevent these reactions, the efficiency of tow molecular mass thiols with an alpha-amino-beta-mercapto-ethane group (Cys, penicillamine and N-acetylcysteine (NAcCys, with a blocked amino group)) as scavengers of methylglyoxal, compared with glutathione (GSH) and the biguanidine derivative metformin, was investigated. The time courses of the reactions of the aforementioned compounds with methylglyoxal were assayed. The reactivity of their thiol and amino groups decreased in the order of Cys  gt  penicillamine  gt  GSH  gt  NAcCys and penicillamine  gt  Cys  gt  GSH, respectively. Human serum albumin (HSA) carbonylation in the absence or presence of methylglyoxal scavengers were monitored by the determination of the amino, guanidine and thiol groups' contents, as well as by spectrofluorimetry, CD and native and SDS PAGE. Cys and penicillamine were highly efficient in the prevention of the carbonylation of the HSA-amino (for 80%) and guanidine (for 84% and 55%, respectively) groups and the formation of fluorescent AGEs. GSH and metformin exhibited medium efficiency (reduction of amino group's carbonylation for 60% and guanidine for about 30%); the least efficient was NAcCys. The presence of Cys, penicillamine and NAcCys led to an almost complete protection of the HSA-thiol group's carbonylation, whereas metformin was inefficient. The efficiency in the prevention of protein cross-linking increased in the order of metformin, NAcCys  lt  GSH  lt  penicillamine  lt  Cys. Thus, the substances with an alpha-amino-beta-mercapto-ethane group as a pharmacophore exhibit great potential as an efficient methylglyoxal scavengers, and are thus promising compounds for medicinal chemistry. In addition, they protect the HSA-SH group and preserve its antioxidative potential, which is very important for the HSA's function in vivo.",
publisher = "Royal Soc Chemistry, Cambridge",
journal = "Molecular BioSystems",
title = "The efficiency of compounds with alpha-amino-beta-mercapto-ethane group in protection of human serum albumin carbonylation and cross-linking with methylglyoxal",
volume = "10",
number = "8",
pages = "2166-2175",
doi = "10.1039/c4mb00217b"
}

Aćimović, J. M., Penezić, A. Z., Pavićević, I. D., Jovanović, V. B.,& Mandić, L. M.. (2014). The efficiency of compounds with alpha-amino-beta-mercapto-ethane group in protection of human serum albumin carbonylation and cross-linking with methylglyoxal. in Molecular BioSystems
Royal Soc Chemistry, Cambridge., 10(8), 2166-2175.
https://doi.org/10.1039/c4mb00217b

Aćimović JM, Penezić AZ, Pavićević ID, Jovanović VB, Mandić LM. The efficiency of compounds with alpha-amino-beta-mercapto-ethane group in protection of human serum albumin carbonylation and cross-linking with methylglyoxal. in Molecular BioSystems. 2014;10(8):2166-2175.
doi:10.1039/c4mb00217b .

Aćimović, Jelena M., Penezić, Ana Z., Pavićević, Ivan D., Jovanović, Vesna B., Mandić, Ljuba M., "The efficiency of compounds with alpha-amino-beta-mercapto-ethane group in protection of human serum albumin carbonylation and cross-linking with methylglyoxal" in Molecular BioSystems, 10, no. 8 (2014):2166-2175,
https://doi.org/10.1039/c4mb00217b . .

TY  - JOUR
AU  - Jovanović, Vesna B.
AU  - Penezić-Romanjuk, Ana Z.
AU  - Pavićević, Ivan D.
AU  - Aćimović, Jelena M.
AU  - Mandić, Ljuba M.
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1369
AB  - The thiol (Cys34) content of human serum albumin (HSA-SH) decreases during oxidative and carbonyl stress and, therefore, could represent a useful parameter in clinical practice. Nevertheless, the reliability of HSA-thiol determination with Ellman's method depends on the purity of isolated HSA. Determination of total serum thiols (mmol/L) and HSA-SH content (mmol -SH/mmol HSA) after HSA isolation from diabetic patient and control sera by a two-step precipitation with ammonium sulfate (AS), as well as HSA-SH contribution (%) to total serum thiols, was assessed. Purity and yield of isolated HSA were monitored spectrophotometrically and by native polyacrylamide gel electrophoresis. Precipitation of HSA from serum via a two-step method with AS produced HSA with 91.9 +/- 3.6% purity and 69.7 +/- 4.4% yield, allowing for precise (relative standard deviation of 3.2%) and reliable (comparing with total serum thiols) measurement of HSA-SH content with DTNB [5,5'-dithiobis-(2-nitrobenzoic acid)]. The content of the HSA-SH group in patients with type 2 diabetes was significantly (P  lt  0.05) lower compared with that of the healthy cohort (0.483 +/- 0.067 vs. 0.561 +/- 0.054 mmol -SH/mmol HSA). Because the proposed method of HSA isolation is simple, time-efficient, and technically less demanding, and it also enables reliable determination of HSA-SH content, it is suitable for clinical practice.
PB  - Academic Press Inc Elsevier Science, San Diego
T2  - Analytical Biochemistry
T1  - Improving the reliability of human serum albumin-thiol group determination
VL  - 439
IS  - 1
SP  - 17
EP  - 22
DO  - 10.1016/j.ab.2013.03.033
ER  - 

@article{
author = "Jovanović, Vesna B. and Penezić-Romanjuk, Ana Z. and Pavićević, Ivan D. and Aćimović, Jelena M. and Mandić, Ljuba M.",
year = "2013",
abstract = "The thiol (Cys34) content of human serum albumin (HSA-SH) decreases during oxidative and carbonyl stress and, therefore, could represent a useful parameter in clinical practice. Nevertheless, the reliability of HSA-thiol determination with Ellman's method depends on the purity of isolated HSA. Determination of total serum thiols (mmol/L) and HSA-SH content (mmol -SH/mmol HSA) after HSA isolation from diabetic patient and control sera by a two-step precipitation with ammonium sulfate (AS), as well as HSA-SH contribution (%) to total serum thiols, was assessed. Purity and yield of isolated HSA were monitored spectrophotometrically and by native polyacrylamide gel electrophoresis. Precipitation of HSA from serum via a two-step method with AS produced HSA with 91.9 +/- 3.6% purity and 69.7 +/- 4.4% yield, allowing for precise (relative standard deviation of 3.2%) and reliable (comparing with total serum thiols) measurement of HSA-SH content with DTNB [5,5'-dithiobis-(2-nitrobenzoic acid)]. The content of the HSA-SH group in patients with type 2 diabetes was significantly (P  lt  0.05) lower compared with that of the healthy cohort (0.483 +/- 0.067 vs. 0.561 +/- 0.054 mmol -SH/mmol HSA). Because the proposed method of HSA isolation is simple, time-efficient, and technically less demanding, and it also enables reliable determination of HSA-SH content, it is suitable for clinical practice.",
publisher = "Academic Press Inc Elsevier Science, San Diego",
journal = "Analytical Biochemistry",
title = "Improving the reliability of human serum albumin-thiol group determination",
volume = "439",
number = "1",
pages = "17-22",
doi = "10.1016/j.ab.2013.03.033"
}

Jovanović, V. B., Penezić-Romanjuk, A. Z., Pavićević, I. D., Aćimović, J. M.,& Mandić, L. M.. (2013). Improving the reliability of human serum albumin-thiol group determination. in Analytical Biochemistry
Academic Press Inc Elsevier Science, San Diego., 439(1), 17-22.
https://doi.org/10.1016/j.ab.2013.03.033

Jovanović VB, Penezić-Romanjuk AZ, Pavićević ID, Aćimović JM, Mandić LM. Improving the reliability of human serum albumin-thiol group determination. in Analytical Biochemistry. 2013;439(1):17-22.
doi:10.1016/j.ab.2013.03.033 .

Jovanović, Vesna B., Penezić-Romanjuk, Ana Z., Pavićević, Ivan D., Aćimović, Jelena M., Mandić, Ljuba M., "Improving the reliability of human serum albumin-thiol group determination" in Analytical Biochemistry, 439, no. 1 (2013):17-22,
https://doi.org/10.1016/j.ab.2013.03.033 . .

TY  - JOUR
AU  - Aćimović, Jelena M.
AU  - Jovanović, Vesna B.
AU  - Sreckovic, Vesna Dimitrijevic
AU  - Penezić-Romanjuk, Ana Z.
AU  - Mandić, Ljuba M.
PY  - 2013
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1578
AB  - Carbonylation of the protein amino, guanidine, and thiol groups with alpha-oxoaldehydes (which are produced in higher quantities in diabetes, uremia, oxidative stress, aging, and inflammation) is one of the important causes of vascular complications. For monitoring of the human serum albumin (HSA) carbonylation level, a spectrophotometric method based on the formation of colored adduct between guanidine group and thymol-sodium hypobromite reagent in the alkaline medium was investigated. Beer's law is obeyed in the concentration range of Arg and protein guanidine groups from 1 to 40 mM. Precision of the method (relative standard deviation) was in the range of 0.9 to 2%. Accuracy was examined by the standard addition method (recovery similar to 100%). The method was applied for monitoring of the carbonylation level of HSA with methylglyoxal in vitro and of HSA isolated (using affinity chromatography) from sera of 21 patients with type 2 diabetes and 12 healthy persons. The content of guanidine groups in HSA isolated from diabetics (19.64 +/- 1.07 mM/mM albumin) was significantly lower (P  lt  0.001) in comparison with a control group (21.87 +/- 1.02 mM/mM albumin). The method is simple and fast, has good accuracy and precision, and is suitable for clinical practice as well for in vitro protein carbonylation experiments.
PB  - Academic Press Inc Elsevier Science, San Diego
T2  - Analytical Biochemistry
T1  - Monitoring of the human serum albumin carbonylation level through determination of guanidino group content
VL  - 433
IS  - 2
SP  - 162
EP  - 167
DO  - 10.1016/j.ab.2012.10.028
ER  - 

@article{
author = "Aćimović, Jelena M. and Jovanović, Vesna B. and Sreckovic, Vesna Dimitrijevic and Penezić-Romanjuk, Ana Z. and Mandić, Ljuba M.",
year = "2013",
abstract = "Carbonylation of the protein amino, guanidine, and thiol groups with alpha-oxoaldehydes (which are produced in higher quantities in diabetes, uremia, oxidative stress, aging, and inflammation) is one of the important causes of vascular complications. For monitoring of the human serum albumin (HSA) carbonylation level, a spectrophotometric method based on the formation of colored adduct between guanidine group and thymol-sodium hypobromite reagent in the alkaline medium was investigated. Beer's law is obeyed in the concentration range of Arg and protein guanidine groups from 1 to 40 mM. Precision of the method (relative standard deviation) was in the range of 0.9 to 2%. Accuracy was examined by the standard addition method (recovery similar to 100%). The method was applied for monitoring of the carbonylation level of HSA with methylglyoxal in vitro and of HSA isolated (using affinity chromatography) from sera of 21 patients with type 2 diabetes and 12 healthy persons. The content of guanidine groups in HSA isolated from diabetics (19.64 +/- 1.07 mM/mM albumin) was significantly lower (P  lt  0.001) in comparison with a control group (21.87 +/- 1.02 mM/mM albumin). The method is simple and fast, has good accuracy and precision, and is suitable for clinical practice as well for in vitro protein carbonylation experiments.",
publisher = "Academic Press Inc Elsevier Science, San Diego",
journal = "Analytical Biochemistry",
title = "Monitoring of the human serum albumin carbonylation level through determination of guanidino group content",
volume = "433",
number = "2",
pages = "162-167",
doi = "10.1016/j.ab.2012.10.028"
}

Aćimović, J. M., Jovanović, V. B., Sreckovic, V. D., Penezić-Romanjuk, A. Z.,& Mandić, L. M.. (2013). Monitoring of the human serum albumin carbonylation level through determination of guanidino group content. in Analytical Biochemistry
Academic Press Inc Elsevier Science, San Diego., 433(2), 162-167.
https://doi.org/10.1016/j.ab.2012.10.028

Aćimović JM, Jovanović VB, Sreckovic VD, Penezić-Romanjuk AZ, Mandić LM. Monitoring of the human serum albumin carbonylation level through determination of guanidino group content. in Analytical Biochemistry. 2013;433(2):162-167.
doi:10.1016/j.ab.2012.10.028 .

Aćimović, Jelena M., Jovanović, Vesna B., Sreckovic, Vesna Dimitrijevic, Penezić-Romanjuk, Ana Z., Mandić, Ljuba M., "Monitoring of the human serum albumin carbonylation level through determination of guanidino group content" in Analytical Biochemistry, 433, no. 2 (2013):162-167,
https://doi.org/10.1016/j.ab.2012.10.028 . .

TY  - THES
AU  - Aćimović, Jelena M.
PY  - 2012
UR  - http://eteze.bg.ac.rs/application/showtheses?thesesId=1137
UR  - https://fedorabg.bg.ac.rs/fedora/get/o:7907/bdef:Content/download
UR  - http://vbs.rs/scripts/cobiss?command=DISPLAY&base=70036&RID=45164047
UR  - http://nardus.mpn.gov.rs/123456789/3490
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2642
AB  - α-Dikarbonilna jedinjenja su veoma reaktivna, reaguju sa nukleofilnim grupamabocnih ostataka Lys i Arg, N-terminalnom amino-grupom i tiol-grupom ostatka Cys napovršini molekula proteina i tako ih modifikuju. U disertaciji je ispitan doprinos tiolgrupemodifikaciji proteina metilglioksalom (MG), kinetika i konkurentnost overeakcije u odnosu na reakcije amino- i guanidino-grupe. Nađeno je da i pored malezastupljenosti tiol-grupe na površni molekula proteina (kod HSA oko 80 puta manja uodnosu na ukupan broj amino- i guanidino-grupa) ona reaguje u velikom procentu (do65 %), ucestvuje u umrežavanju molekula proteina (stvaranju dimera i oligomera) prekonagrađenih HSA-MG (HSA-amino-MG ili HSA-SH-MG) intermedijera sa udelom od4% u slucaju nedovoljne kolicine metilglioksala Povecanje koncentracije metilglioksalautice na brzinu reakcije sa tiol-grupom, na vreme uspostavljanja ravnoteže, na doprinostiol-grupe modifikaciji molekula proteina, ali ne utiče na procenat izreagovanih grupa ustanju ravnoteže (60 %), što omogucava ekstrapolaciju dobijenih rezultata na niže,fiziološke koncentracije metilglioksala.U okviru teze su razvijene spektrofotometrijske metode zasnovane na reakcijiamino-grupa sa p-benzohinonom i guanidino-grupa sa reagensom koji sadrži timol inatrijum-hipobromit. Metode su precizne (RSD u opsegu 1.2-1.8 %, odnosno 0.9–2.0%, resp.) i tačne (100.6 ± 1.2 %, 99.8 ± 0.8 %, redom), što ih cini pogodnim zakvantifikaciju ovih grupa za vreme karbonilovanja in vitro.Uticaj mikrookoline tiol-grupe na njenu reakciju sa metilglioksalom ispitan je saaminokiselinama (Cys, NAcCys i CMC), peptidom (GSH) i proteinom (HSA).Utvrđeno je da mikrookolina –SH grupe utice na njenu reaktivnost i na prinos proizvodareakcije. Reaktivnost tiol-grupe opada u nizu Cys>GSH>NAcCys, za sve ispitivaneodnose koncentracija reaktanata. CMC ne reaguje sa metilglioksalom. Prisustvo α-amino-β-merkapto-etanske grupe u tiol-jedinjenju povecava reaktivnost tiol-grupe.Predožen je reakcioni put za reakciju Cys i metilglioksala po kome slobodna α-amino-grupa molekula Cys (cija je tiol-grupa u reakciji sa metilglioksalom vecnagradila hemitioacetal), reaguje sa slobodnom acetil-karbonilnom grupomhemitioacetala. Stvara se tiazolidinski intermedijer, koji dalje prelazi u stabilniji proizvod Cys-MG-glikozilamin...
AB  - α-Dicarbonyles are reactive compounds that react with the nucleophilic groupsof Lys and Arg side chains, thiol group of Cys and N-terminal amino on the proteinsurface, modifying them. In this thesis, the contribution of thiol group to proteinmodification with methylglyoxal (MG), kinetics and competitivness of this reactionwith those of amino and guanidino groups reactions were inestigated. Although lessabundant, thiol groups present on the protein surface (approx. 80 times lower then totalnumber of amino and guanidino groups in HSA), are found to be reactive inconsiderable percentage (up to 65 %) and are participated in protein cross-linking(dimer and oligomer production), through HSA-MG intermediates (HSA-amino-MG orHSA-SH-MG), with 4% in case of unsufficient amount of MG. Increase inmethylglyoxal concentration influences the reaction rate with thiol group, the time forthe reaction to reach equilibrium and the contribution of thiol group to modification ofproteins, yet it has no influence on the percent of reacted groups (60 %) when chemicalequilibrium is reached, enabling extrapolation of the obtained results to lower,physiological concentrations of methylglyoxal.Spectrophotometric methods, based on the reaction of amino groups with pbenzoquinoneand guanidino groups with thimol and sodium hypobromite reagent, havebeen developed. The methods are precise (RSD 1.2-1.8 % and 0.9–2.0 %, respectively)and accurate (100.6±1.2 %; 99.8±0.8 %, resp.), and therefore suitable for quantificationof these groups during carbonylation process in vitro.The influence of microenviroment of thiol group on its reactivity with MG hasbeen examined using amino acids (Cys, NAcCys and CMC), peptide (GSH) and protein(HSA). It has been shown that microenviroment of thiol group influences its reactivityand the product yield. The reactivity of thiol group decreases in the orderCys>GSH>NAcCys, for all concentration ratios examined. CMC doesn’t react withMG. The presence of α-amino-β-mercapto ethane group in thiol compound increases thereactivity of thiol group.The reaction pathway for Cys and MG reaction was proposed, in which the freeα-amino group of Cys (whose thiol group is already in the form of hemithioacetale with MG) reacts with the free acetil carbonyl group of hemithioacetale...
PB  - Универзитет у Београду, Хемијски факултет
T2  - Универзитет у Београду
T1  - Modifikacija -SH grupe proteina α- dikarbonilnim jedinjenjima : identifikacija proizvoda, mogućnosti određivanja i primene u kliničkoj praksi
T1  - Modification of protein - SH group with α-dicarbonyl compounds: identification of products, the possibilities of determination and application in clinical practice
UR  - https://hdl.handle.net/21.15107/rcub_nardus_3490
ER  - 

@phdthesis{
author = "Aćimović, Jelena M.",
year = "2012",
abstract = "α-Dikarbonilna jedinjenja su veoma reaktivna, reaguju sa nukleofilnim grupamabocnih ostataka Lys i Arg, N-terminalnom amino-grupom i tiol-grupom ostatka Cys napovršini molekula proteina i tako ih modifikuju. U disertaciji je ispitan doprinos tiolgrupemodifikaciji proteina metilglioksalom (MG), kinetika i konkurentnost overeakcije u odnosu na reakcije amino- i guanidino-grupe. Nađeno je da i pored malezastupljenosti tiol-grupe na površni molekula proteina (kod HSA oko 80 puta manja uodnosu na ukupan broj amino- i guanidino-grupa) ona reaguje u velikom procentu (do65 %), ucestvuje u umrežavanju molekula proteina (stvaranju dimera i oligomera) prekonagrađenih HSA-MG (HSA-amino-MG ili HSA-SH-MG) intermedijera sa udelom od4% u slucaju nedovoljne kolicine metilglioksala Povecanje koncentracije metilglioksalautice na brzinu reakcije sa tiol-grupom, na vreme uspostavljanja ravnoteže, na doprinostiol-grupe modifikaciji molekula proteina, ali ne utiče na procenat izreagovanih grupa ustanju ravnoteže (60 %), što omogucava ekstrapolaciju dobijenih rezultata na niže,fiziološke koncentracije metilglioksala.U okviru teze su razvijene spektrofotometrijske metode zasnovane na reakcijiamino-grupa sa p-benzohinonom i guanidino-grupa sa reagensom koji sadrži timol inatrijum-hipobromit. Metode su precizne (RSD u opsegu 1.2-1.8 %, odnosno 0.9–2.0%, resp.) i tačne (100.6 ± 1.2 %, 99.8 ± 0.8 %, redom), što ih cini pogodnim zakvantifikaciju ovih grupa za vreme karbonilovanja in vitro.Uticaj mikrookoline tiol-grupe na njenu reakciju sa metilglioksalom ispitan je saaminokiselinama (Cys, NAcCys i CMC), peptidom (GSH) i proteinom (HSA).Utvrđeno je da mikrookolina –SH grupe utice na njenu reaktivnost i na prinos proizvodareakcije. Reaktivnost tiol-grupe opada u nizu Cys>GSH>NAcCys, za sve ispitivaneodnose koncentracija reaktanata. CMC ne reaguje sa metilglioksalom. Prisustvo α-amino-β-merkapto-etanske grupe u tiol-jedinjenju povecava reaktivnost tiol-grupe.Predožen je reakcioni put za reakciju Cys i metilglioksala po kome slobodna α-amino-grupa molekula Cys (cija je tiol-grupa u reakciji sa metilglioksalom vecnagradila hemitioacetal), reaguje sa slobodnom acetil-karbonilnom grupomhemitioacetala. Stvara se tiazolidinski intermedijer, koji dalje prelazi u stabilniji proizvod Cys-MG-glikozilamin..., α-Dicarbonyles are reactive compounds that react with the nucleophilic groupsof Lys and Arg side chains, thiol group of Cys and N-terminal amino on the proteinsurface, modifying them. In this thesis, the contribution of thiol group to proteinmodification with methylglyoxal (MG), kinetics and competitivness of this reactionwith those of amino and guanidino groups reactions were inestigated. Although lessabundant, thiol groups present on the protein surface (approx. 80 times lower then totalnumber of amino and guanidino groups in HSA), are found to be reactive inconsiderable percentage (up to 65 %) and are participated in protein cross-linking(dimer and oligomer production), through HSA-MG intermediates (HSA-amino-MG orHSA-SH-MG), with 4% in case of unsufficient amount of MG. Increase inmethylglyoxal concentration influences the reaction rate with thiol group, the time forthe reaction to reach equilibrium and the contribution of thiol group to modification ofproteins, yet it has no influence on the percent of reacted groups (60 %) when chemicalequilibrium is reached, enabling extrapolation of the obtained results to lower,physiological concentrations of methylglyoxal.Spectrophotometric methods, based on the reaction of amino groups with pbenzoquinoneand guanidino groups with thimol and sodium hypobromite reagent, havebeen developed. The methods are precise (RSD 1.2-1.8 % and 0.9–2.0 %, respectively)and accurate (100.6±1.2 %; 99.8±0.8 %, resp.), and therefore suitable for quantificationof these groups during carbonylation process in vitro.The influence of microenviroment of thiol group on its reactivity with MG hasbeen examined using amino acids (Cys, NAcCys and CMC), peptide (GSH) and protein(HSA). It has been shown that microenviroment of thiol group influences its reactivityand the product yield. The reactivity of thiol group decreases in the orderCys>GSH>NAcCys, for all concentration ratios examined. CMC doesn’t react withMG. The presence of α-amino-β-mercapto ethane group in thiol compound increases thereactivity of thiol group.The reaction pathway for Cys and MG reaction was proposed, in which the freeα-amino group of Cys (whose thiol group is already in the form of hemithioacetale with MG) reacts with the free acetil carbonyl group of hemithioacetale...",
publisher = "Универзитет у Београду, Хемијски факултет",
journal = "Универзитет у Београду",
title = "Modifikacija -SH grupe proteina α- dikarbonilnim jedinjenjima : identifikacija proizvoda, mogućnosti određivanja i primene u kliničkoj praksi, Modification of protein - SH group with α-dicarbonyl compounds: identification of products, the possibilities of determination and application in clinical practice",
url = "https://hdl.handle.net/21.15107/rcub_nardus_3490"
}

Aćimović, J. M.. (2012). Modifikacija -SH grupe proteina α- dikarbonilnim jedinjenjima : identifikacija proizvoda, mogućnosti određivanja i primene u kliničkoj praksi. in Универзитет у Београду
Универзитет у Београду, Хемијски факултет..
https://hdl.handle.net/21.15107/rcub_nardus_3490

Aćimović JM. Modifikacija -SH grupe proteina α- dikarbonilnim jedinjenjima : identifikacija proizvoda, mogućnosti određivanja i primene u kliničkoj praksi. in Универзитет у Београду. 2012;.
https://hdl.handle.net/21.15107/rcub_nardus_3490 .

Aćimović, Jelena M., "Modifikacija -SH grupe proteina α- dikarbonilnim jedinjenjima : identifikacija proizvoda, mogućnosti određivanja i primene u kliničkoj praksi" in Универзитет у Београду (2012),
https://hdl.handle.net/21.15107/rcub_nardus_3490 .

TY  - JOUR
AU  - Aćimović, Jelena M.
AU  - Jovanović, Vesna B.
AU  - Veselinovic, Milica R.
AU  - Serckovic, Vesna Dimitrijevic
AU  - Mandić, Ljuba M.
PY  - 2011
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1179
AB  - Objectives: Carbonylation of the protein amino, guanidino and thiol groups inane of the important causes of vascular complications in diabetes. We developed a simple spectrophotometric method for monitoring of the changes in the protein amino group contents during carbonylation. Design and methods: The method is based on the reaction of amino group with p-benzoquinone in the slightly alkaline media. It was applied during carbonylation in vitro with methylglyoxal and in vivo in 13 patients with type 2 diabetes and 20 healthy persons. Results: The method is simple, fast, precise (RSD in the range of 1.2-1.8%) and accurate (recovery about 100%). The content of amino groups in human serum albumin isolated from diabetics was significantly lower (p lt 0.01) in comparison with a control group. Conclusion: The method developed is suitable for quantification of protein amino groups during in vitro carbonylation as well as for clinical practice. (C) 2011 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.
PB  - Pergamon-Elsevier Science Ltd, Oxford
T2  - Clinical Biochemistry
T1  - Method for monitoring of the protein amino group changes during carbonylation
VL  - 44
IS  - 12
SP  - 994
EP  - 999
DO  - 10.1016/j.clinbiochem.2011.05.019
ER  - 

@article{
author = "Aćimović, Jelena M. and Jovanović, Vesna B. and Veselinovic, Milica R. and Serckovic, Vesna Dimitrijevic and Mandić, Ljuba M.",
year = "2011",
abstract = "Objectives: Carbonylation of the protein amino, guanidino and thiol groups inane of the important causes of vascular complications in diabetes. We developed a simple spectrophotometric method for monitoring of the changes in the protein amino group contents during carbonylation. Design and methods: The method is based on the reaction of amino group with p-benzoquinone in the slightly alkaline media. It was applied during carbonylation in vitro with methylglyoxal and in vivo in 13 patients with type 2 diabetes and 20 healthy persons. Results: The method is simple, fast, precise (RSD in the range of 1.2-1.8%) and accurate (recovery about 100%). The content of amino groups in human serum albumin isolated from diabetics was significantly lower (p lt 0.01) in comparison with a control group. Conclusion: The method developed is suitable for quantification of protein amino groups during in vitro carbonylation as well as for clinical practice. (C) 2011 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.",
publisher = "Pergamon-Elsevier Science Ltd, Oxford",
journal = "Clinical Biochemistry",
title = "Method for monitoring of the protein amino group changes during carbonylation",
volume = "44",
number = "12",
pages = "994-999",
doi = "10.1016/j.clinbiochem.2011.05.019"
}

Aćimović, J. M., Jovanović, V. B., Veselinovic, M. R., Serckovic, V. D.,& Mandić, L. M.. (2011). Method for monitoring of the protein amino group changes during carbonylation. in Clinical Biochemistry
Pergamon-Elsevier Science Ltd, Oxford., 44(12), 994-999.
https://doi.org/10.1016/j.clinbiochem.2011.05.019

Aćimović JM, Jovanović VB, Veselinovic MR, Serckovic VD, Mandić LM. Method for monitoring of the protein amino group changes during carbonylation. in Clinical Biochemistry. 2011;44(12):994-999.
doi:10.1016/j.clinbiochem.2011.05.019 .

Aćimović, Jelena M., Jovanović, Vesna B., Veselinovic, Milica R., Serckovic, Vesna Dimitrijevic, Mandić, Ljuba M., "Method for monitoring of the protein amino group changes during carbonylation" in Clinical Biochemistry, 44, no. 12 (2011):994-999,
https://doi.org/10.1016/j.clinbiochem.2011.05.019 . .

TY  - JOUR
AU  - Aćimović, Jelena M.
AU  - Stanimirović, Bojana
AU  - Todorović, Nina
AU  - Jovanović, Vesna B.
AU  - Mandić, Ljuba M.
PY  - 2010
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1122
AB  - Methylglyoxal (MG), a reactive alpha-oxoaldehyde that is produced in higher quantities in diabetes, uremia, oxidative stress, aging and inflammation, reacts with the thiol groups (in addition to the amino and guanidino groups) of proteins. This causes protein modification, formation of advanced glycated end products (AGEs) and cross-linking. Low molecular mass thiols can be used as competitive targets for MG, preventing the reactions mentioned above. Therefore, this paper investigated how the microenvironment of the thiol group in low molecular mass thiols (cysteine, N-acetylcysteine (NAcCys), carboxymethylcysteine (CMC) and glutathione (GSH)) and human serum albumin (HSA) affected the thiol reaction with MG. The SH group reaction course was monitored by (1)H-NMR spectroscopy and spectrophotometric quantification. Changes in the HSA molecules were monitored by SDS-PAGE. The microenvironment of the SH group had a major effect on its reactivity and on the product yield. The reactivity of SH groups decreased in the order Cys  gt  GSH  gt  NAcCys. CMC did not react. The percentages of the reacted SH groups in the equilibrium state were almost equal, regardless of the ratio of thiol compound/MG (1:1, 1:2, 1:5): 38.1 +/- 0.9%; 38.2 +/- 0.7% and 39.0 +/- 0.8% for Cys; 26.5 +/- 0.6%; 26.6 +/- 2.6% and 27.4 +/- 2.5% for GSH; 10.8 +/- 0.9%; and 11.2 +/- 0.7% and 12.2 +/- 0.9% for NAcCys, respectively. Our results explain why substances containing alpha-amino-beta-mercapto-ethane as a pharmacophore are successful scavengers of MG. In equilibrium, HSA SH reacted in high percentages both with an insufficient amount and with an excess of MG (55% and 65%, respectively). An analysis of the hydrophobicity of the microenvironment of the SH group on the HSA surface showed that it could contribute to high levels of SH modification, leading to an increase in the scavenging activity of the albumin thiol. (C) 2010 Elsevier Ireland Ltd. All rights reserved.
PB  - Elsevier Ireland Ltd, Clare
T2  - Chemico-biological Interactions
T1  - Influence of the microenvironment of thiol groups in low molecular mass thiols and serum albumin on the reaction with methylglyoxal
VL  - 188
IS  - 1
SP  - 21
EP  - 30
DO  - 10.1016/j.cbi.2010.07.013
ER  - 

@article{
author = "Aćimović, Jelena M. and Stanimirović, Bojana and Todorović, Nina and Jovanović, Vesna B. and Mandić, Ljuba M.",
year = "2010",
abstract = "Methylglyoxal (MG), a reactive alpha-oxoaldehyde that is produced in higher quantities in diabetes, uremia, oxidative stress, aging and inflammation, reacts with the thiol groups (in addition to the amino and guanidino groups) of proteins. This causes protein modification, formation of advanced glycated end products (AGEs) and cross-linking. Low molecular mass thiols can be used as competitive targets for MG, preventing the reactions mentioned above. Therefore, this paper investigated how the microenvironment of the thiol group in low molecular mass thiols (cysteine, N-acetylcysteine (NAcCys), carboxymethylcysteine (CMC) and glutathione (GSH)) and human serum albumin (HSA) affected the thiol reaction with MG. The SH group reaction course was monitored by (1)H-NMR spectroscopy and spectrophotometric quantification. Changes in the HSA molecules were monitored by SDS-PAGE. The microenvironment of the SH group had a major effect on its reactivity and on the product yield. The reactivity of SH groups decreased in the order Cys  gt  GSH  gt  NAcCys. CMC did not react. The percentages of the reacted SH groups in the equilibrium state were almost equal, regardless of the ratio of thiol compound/MG (1:1, 1:2, 1:5): 38.1 +/- 0.9%; 38.2 +/- 0.7% and 39.0 +/- 0.8% for Cys; 26.5 +/- 0.6%; 26.6 +/- 2.6% and 27.4 +/- 2.5% for GSH; 10.8 +/- 0.9%; and 11.2 +/- 0.7% and 12.2 +/- 0.9% for NAcCys, respectively. Our results explain why substances containing alpha-amino-beta-mercapto-ethane as a pharmacophore are successful scavengers of MG. In equilibrium, HSA SH reacted in high percentages both with an insufficient amount and with an excess of MG (55% and 65%, respectively). An analysis of the hydrophobicity of the microenvironment of the SH group on the HSA surface showed that it could contribute to high levels of SH modification, leading to an increase in the scavenging activity of the albumin thiol. (C) 2010 Elsevier Ireland Ltd. All rights reserved.",
publisher = "Elsevier Ireland Ltd, Clare",
journal = "Chemico-biological Interactions",
title = "Influence of the microenvironment of thiol groups in low molecular mass thiols and serum albumin on the reaction with methylglyoxal",
volume = "188",
number = "1",
pages = "21-30",
doi = "10.1016/j.cbi.2010.07.013"
}

Aćimović, J. M., Stanimirović, B., Todorović, N., Jovanović, V. B.,& Mandić, L. M.. (2010). Influence of the microenvironment of thiol groups in low molecular mass thiols and serum albumin on the reaction with methylglyoxal. in Chemico-biological Interactions
Elsevier Ireland Ltd, Clare., 188(1), 21-30.
https://doi.org/10.1016/j.cbi.2010.07.013

Aćimović JM, Stanimirović B, Todorović N, Jovanović VB, Mandić LM. Influence of the microenvironment of thiol groups in low molecular mass thiols and serum albumin on the reaction with methylglyoxal. in Chemico-biological Interactions. 2010;188(1):21-30.
doi:10.1016/j.cbi.2010.07.013 .

Aćimović, Jelena M., Stanimirović, Bojana, Todorović, Nina, Jovanović, Vesna B., Mandić, Ljuba M., "Influence of the microenvironment of thiol groups in low molecular mass thiols and serum albumin on the reaction with methylglyoxal" in Chemico-biological Interactions, 188, no. 1 (2010):21-30,
https://doi.org/10.1016/j.cbi.2010.07.013 . .

TY  - JOUR
AU  - Aćimović, Jelena M.
AU  - Stanimirović, Bojana
AU  - Mandić, Ljuba M.
PY  - 2009
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1015
AB  - Methylglyoxal is a highly reactive a-oxoaldehyde with elevated production in hyperglycemia. It reacts with nucleophilic Lys and Arg side-chains and N-terminal amino groups causing protein modification. In the present study, the importance of the reaction of the Cys thiol group with methylglyoxal in protein modification, the competitiveness of this reaction with those of amino and guanidine groups, the time course of these reactions and their role and contribution to protein cross-linking were investigated. Human and bovine serum albumins were used as model systems. It was found that despite the very low levels of thiol groups on the surface of the examined protein molecules (approx. 80 times lower than those of amino and guanidino groups), a very high percentage of it reacts (25-85%). The amount of reacted thiol groups and the rate of the reaction, the time for the reaction to reach equilibrium, the formation of a stable product and the contribution of thiol groups to protein cross-linking depend on the methylglyoxal concentration. The product formed in the reaction of thiol and an insufficient quantity of methylglyoxal (compared to the concentrations of the groups accessible for modification) participates to a significant extent (4%) to protein cross-linking. Metformin applied in equimolar concentration with methylglyoxal prevents its reaction with amino and guanidino groups but, however, not with thiol groups.
AB  - Metilglioksal je veoma reaktivni α-oksoaldehid koji se povećano stvara u hiperglikemiji. Reaguje sa nukleofilnim grupama bočnih ostataka Lys, Arg i N-terminalnom amino-grupom, što dovodi do modifikacije proteina. U ovom radu ispitivani su značaj reakcije SH grupe sa metilglioksalom u modifikaciji proteina, konkurentnost ove reakcije u odnosu na reakcije sa amino- i gvanidino-grupom, tok ovih reakcija i njihova uloga i doprinos u umrežavanju proteina. Kao model-sistemi upotrebljeni su humani i goveđi serum-albumin. Utvrđeno je da i pored veoma male zastupljenosti SH grupe na površini ispitivanih molekula proteina (oko 80 puta manja u odnosu na ukupan broj amino- i gvanidino-grupa), ona reaguje u velikom procentu (od 25-85 %). Količina izreagovanih SH grupa i brzina reakcije, vreme uspostavljanja ravnoteže reakcije, stvaranja stabilnog proizvoda i doprinos SH grupa umrežavanju proteina zavise od koncentracije metilglioksala. Proizvod stvoren u reakciji SH grupa i nedovoljne količine metilglioksala (u odnosu na koncentraciju grupa dostupnih za modifikaciju) učestvuje u umrežavanju proteina sa značajnim udelom (4 %). U ekvimolarnoj koncentraciji sa metilglioksalom metformin sprečava njegovu reakciju sa amino i guanidino grupama albumina, ali ne i sa tiol grupom.
PB  - Serbian Chemical Soc, Belgrade
T2  - Journal of the Serbian Chemical Society
T1  - The role of the thiol group in protein modification with methylglyoxal
T1  - Uloga tiolne grupe u modifikaciji proteina sa metilglioksalom
VL  - 74
IS  - 8-9
SP  - 867
EP  - 883
DO  - 10.2298/JSC0909867A
ER  - 

@article{
author = "Aćimović, Jelena M. and Stanimirović, Bojana and Mandić, Ljuba M.",
year = "2009",
abstract = "Methylglyoxal is a highly reactive a-oxoaldehyde with elevated production in hyperglycemia. It reacts with nucleophilic Lys and Arg side-chains and N-terminal amino groups causing protein modification. In the present study, the importance of the reaction of the Cys thiol group with methylglyoxal in protein modification, the competitiveness of this reaction with those of amino and guanidine groups, the time course of these reactions and their role and contribution to protein cross-linking were investigated. Human and bovine serum albumins were used as model systems. It was found that despite the very low levels of thiol groups on the surface of the examined protein molecules (approx. 80 times lower than those of amino and guanidino groups), a very high percentage of it reacts (25-85%). The amount of reacted thiol groups and the rate of the reaction, the time for the reaction to reach equilibrium, the formation of a stable product and the contribution of thiol groups to protein cross-linking depend on the methylglyoxal concentration. The product formed in the reaction of thiol and an insufficient quantity of methylglyoxal (compared to the concentrations of the groups accessible for modification) participates to a significant extent (4%) to protein cross-linking. Metformin applied in equimolar concentration with methylglyoxal prevents its reaction with amino and guanidino groups but, however, not with thiol groups., Metilglioksal je veoma reaktivni α-oksoaldehid koji se povećano stvara u hiperglikemiji. Reaguje sa nukleofilnim grupama bočnih ostataka Lys, Arg i N-terminalnom amino-grupom, što dovodi do modifikacije proteina. U ovom radu ispitivani su značaj reakcije SH grupe sa metilglioksalom u modifikaciji proteina, konkurentnost ove reakcije u odnosu na reakcije sa amino- i gvanidino-grupom, tok ovih reakcija i njihova uloga i doprinos u umrežavanju proteina. Kao model-sistemi upotrebljeni su humani i goveđi serum-albumin. Utvrđeno je da i pored veoma male zastupljenosti SH grupe na površini ispitivanih molekula proteina (oko 80 puta manja u odnosu na ukupan broj amino- i gvanidino-grupa), ona reaguje u velikom procentu (od 25-85 %). Količina izreagovanih SH grupa i brzina reakcije, vreme uspostavljanja ravnoteže reakcije, stvaranja stabilnog proizvoda i doprinos SH grupa umrežavanju proteina zavise od koncentracije metilglioksala. Proizvod stvoren u reakciji SH grupa i nedovoljne količine metilglioksala (u odnosu na koncentraciju grupa dostupnih za modifikaciju) učestvuje u umrežavanju proteina sa značajnim udelom (4 %). U ekvimolarnoj koncentraciji sa metilglioksalom metformin sprečava njegovu reakciju sa amino i guanidino grupama albumina, ali ne i sa tiol grupom.",
publisher = "Serbian Chemical Soc, Belgrade",
journal = "Journal of the Serbian Chemical Society",
title = "The role of the thiol group in protein modification with methylglyoxal, Uloga tiolne grupe u modifikaciji proteina sa metilglioksalom",
volume = "74",
number = "8-9",
pages = "867-883",
doi = "10.2298/JSC0909867A"
}

Aćimović, J. M., Stanimirović, B.,& Mandić, L. M.. (2009). The role of the thiol group in protein modification with methylglyoxal. in Journal of the Serbian Chemical Society
Serbian Chemical Soc, Belgrade., 74(8-9), 867-883.
https://doi.org/10.2298/JSC0909867A

Aćimović JM, Stanimirović B, Mandić LM. The role of the thiol group in protein modification with methylglyoxal. in Journal of the Serbian Chemical Society. 2009;74(8-9):867-883.
doi:10.2298/JSC0909867A .

Aćimović, Jelena M., Stanimirović, Bojana, Mandić, Ljuba M., "The role of the thiol group in protein modification with methylglyoxal" in Journal of the Serbian Chemical Society, 74, no. 8-9 (2009):867-883,
https://doi.org/10.2298/JSC0909867A . .

TY  - CONF
AU  - Ivanovska, J.
AU  - Mladenovic, S.
AU  - Nastasijevic, B.
AU  - Miletić, S.
AU  - Aćimović, Jelena M.
PY  - 2007
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/915
PB  - Blackwell Publishing, Oxford
C3  - FEBS Journal / Federation of European of Biochemical Societies
T1  - Efficiency of commercial enzymatic preparations in grain hydrolysis for preparation of substrates for beer fermentation
VL  - 274
SP  - 224
EP  - 224
UR  - https://hdl.handle.net/21.15107/rcub_cherry_915
ER  - 

@conference{
author = "Ivanovska, J. and Mladenovic, S. and Nastasijevic, B. and Miletić, S. and Aćimović, Jelena M.",
year = "2007",
publisher = "Blackwell Publishing, Oxford",
journal = "FEBS Journal / Federation of European of Biochemical Societies",
title = "Efficiency of commercial enzymatic preparations in grain hydrolysis for preparation of substrates for beer fermentation",
volume = "274",
pages = "224-224",
url = "https://hdl.handle.net/21.15107/rcub_cherry_915"
}

Ivanovska, J., Mladenovic, S., Nastasijevic, B., Miletić, S.,& Aćimović, J. M.. (2007). Efficiency of commercial enzymatic preparations in grain hydrolysis for preparation of substrates for beer fermentation. in FEBS Journal / Federation of European of Biochemical Societies
Blackwell Publishing, Oxford., 274, 224-224.
https://hdl.handle.net/21.15107/rcub_cherry_915

Ivanovska J, Mladenovic S, Nastasijevic B, Miletić S, Aćimović JM. Efficiency of commercial enzymatic preparations in grain hydrolysis for preparation of substrates for beer fermentation. in FEBS Journal / Federation of European of Biochemical Societies. 2007;274:224-224.
https://hdl.handle.net/21.15107/rcub_cherry_915 .

Ivanovska, J., Mladenovic, S., Nastasijevic, B., Miletić, S., Aćimović, Jelena M., "Efficiency of commercial enzymatic preparations in grain hydrolysis for preparation of substrates for beer fermentation" in FEBS Journal / Federation of European of Biochemical Societies, 274 (2007):224-224,
https://hdl.handle.net/21.15107/rcub_cherry_915 .

TY  - CONF
AU  - Jankovic, J.
AU  - Aćimović, Jelena M.
AU  - Stanimirović, Bojana
AU  - Ivanovska, J.
AU  - Mandić, Ljuba M.
PY  - 2007
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/916
PB  - Blackwell Publishing, Oxford
C3  - FEBS Journal / Federation of European of Biochemical Societies
T1  - Biochemical characterization of advanced glycation end products: Modifications of thiol amino-acid residue
VL  - 274
SP  - 241
EP  - 241
UR  - https://hdl.handle.net/21.15107/rcub_cherry_916
ER  - 

@conference{
author = "Jankovic, J. and Aćimović, Jelena M. and Stanimirović, Bojana and Ivanovska, J. and Mandić, Ljuba M.",
year = "2007",
publisher = "Blackwell Publishing, Oxford",
journal = "FEBS Journal / Federation of European of Biochemical Societies",
title = "Biochemical characterization of advanced glycation end products: Modifications of thiol amino-acid residue",
volume = "274",
pages = "241-241",
url = "https://hdl.handle.net/21.15107/rcub_cherry_916"
}

Jankovic, J., Aćimović, J. M., Stanimirović, B., Ivanovska, J.,& Mandić, L. M.. (2007). Biochemical characterization of advanced glycation end products: Modifications of thiol amino-acid residue. in FEBS Journal / Federation of European of Biochemical Societies
Blackwell Publishing, Oxford., 274, 241-241.
https://hdl.handle.net/21.15107/rcub_cherry_916

Jankovic J, Aćimović JM, Stanimirović B, Ivanovska J, Mandić LM. Biochemical characterization of advanced glycation end products: Modifications of thiol amino-acid residue. in FEBS Journal / Federation of European of Biochemical Societies. 2007;274:241-241.
https://hdl.handle.net/21.15107/rcub_cherry_916 .

Jankovic, J., Aćimović, Jelena M., Stanimirović, Bojana, Ivanovska, J., Mandić, Ljuba M., "Biochemical characterization of advanced glycation end products: Modifications of thiol amino-acid residue" in FEBS Journal / Federation of European of Biochemical Societies, 274 (2007):241-241,
https://hdl.handle.net/21.15107/rcub_cherry_916 .

TY  - CONF
AU  - Ivanovska, J.
AU  - Jankovic, J.
AU  - Aćimović, Jelena M.
AU  - Stanimirović, Bojana
AU  - Mandić, Ljuba M.
PY  - 2007
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/917
PB  - Blackwell Publishing, Oxford
C3  - FEBS Journal / Federation of European of Biochemical Societies
T1  - Kinetics of reaction of human serum albumine with methyl-glyoxal
VL  - 274
SP  - 241
EP  - 241
UR  - https://hdl.handle.net/21.15107/rcub_cherry_917
ER  - 

@conference{
author = "Ivanovska, J. and Jankovic, J. and Aćimović, Jelena M. and Stanimirović, Bojana and Mandić, Ljuba M.",
year = "2007",
publisher = "Blackwell Publishing, Oxford",
journal = "FEBS Journal / Federation of European of Biochemical Societies",
title = "Kinetics of reaction of human serum albumine with methyl-glyoxal",
volume = "274",
pages = "241-241",
url = "https://hdl.handle.net/21.15107/rcub_cherry_917"
}

Ivanovska, J., Jankovic, J., Aćimović, J. M., Stanimirović, B.,& Mandić, L. M.. (2007). Kinetics of reaction of human serum albumine with methyl-glyoxal. in FEBS Journal / Federation of European of Biochemical Societies
Blackwell Publishing, Oxford., 274, 241-241.
https://hdl.handle.net/21.15107/rcub_cherry_917

Ivanovska J, Jankovic J, Aćimović JM, Stanimirović B, Mandić LM. Kinetics of reaction of human serum albumine with methyl-glyoxal. in FEBS Journal / Federation of European of Biochemical Societies. 2007;274:241-241.
https://hdl.handle.net/21.15107/rcub_cherry_917 .

Ivanovska, J., Jankovic, J., Aćimović, Jelena M., Stanimirović, Bojana, Mandić, Ljuba M., "Kinetics of reaction of human serum albumine with methyl-glyoxal" in FEBS Journal / Federation of European of Biochemical Societies, 274 (2007):241-241,
https://hdl.handle.net/21.15107/rcub_cherry_917 .

TY  - JOUR
AU  - Aćimović, Jelena M.
AU  - Jovanović, Vesna B.
AU  - Mandić, Ljuba M.
PY  - 2005
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/527
AB  - The influence of urinary pigments and urine pH on the spectrophotometric determination of N-acetyl-beta-D-glucosaminidase (NAG; EC 3.2.1.30) activity with 2-methoxy-4-(2'-nitrovinyl)-phenyl-N-acetyl-beta-D-glucosaminide as a substrate was studied. The investigation was performed with human and rabbit urine samples. It was found that alkaline urine pH values influenced NAG activity in two ways: 1) NAG activity decreased due to enzyme instability with pH increase, and 2) NAG activity increased because of the contribution of urinary pigments to absorbance of 2-methoxy-4-(2'-nitrovinyl)-phenol (MNP) at 505nm. It was shown that besides the maximum (1) in the range of 350-360nm of the absorption spectra of alkaline urine, there was a maximum (II) in the range of 380-460nm. With the increase of pH, maximum 11 was shifted toward higher wavelengths and contributed to MNP absorption (5-90%). On the other hand, the maximum of MNP absorption was shifted toward lower wavelengths (495-400nm) with increasing pH. Two procedures to eliminate the influence of urinary pigments are presented. The justification of applying a correction to the values of NAG activity in human and rabbit urine (a model system for studying the toxic effects of cadmium) was discussed.
PB  - Wiley-Liss, Hoboken
T2  - Journal of Clinical Laboratory Analysis
T1  - Influence of pigments and pH of urine on the determination of N-acetyl-beta-D-glucosaminidase activity with 2-methoxy-4-(2 '-nitrovinyl)-phenyl-N-acetyl-beta-D-glucosaminide
VL  - 19
IS  - 6
SP  - 260
EP  - 266
DO  - 10.1002/jcla.20088
ER  - 

@article{
author = "Aćimović, Jelena M. and Jovanović, Vesna B. and Mandić, Ljuba M.",
year = "2005",
abstract = "The influence of urinary pigments and urine pH on the spectrophotometric determination of N-acetyl-beta-D-glucosaminidase (NAG; EC 3.2.1.30) activity with 2-methoxy-4-(2'-nitrovinyl)-phenyl-N-acetyl-beta-D-glucosaminide as a substrate was studied. The investigation was performed with human and rabbit urine samples. It was found that alkaline urine pH values influenced NAG activity in two ways: 1) NAG activity decreased due to enzyme instability with pH increase, and 2) NAG activity increased because of the contribution of urinary pigments to absorbance of 2-methoxy-4-(2'-nitrovinyl)-phenol (MNP) at 505nm. It was shown that besides the maximum (1) in the range of 350-360nm of the absorption spectra of alkaline urine, there was a maximum (II) in the range of 380-460nm. With the increase of pH, maximum 11 was shifted toward higher wavelengths and contributed to MNP absorption (5-90%). On the other hand, the maximum of MNP absorption was shifted toward lower wavelengths (495-400nm) with increasing pH. Two procedures to eliminate the influence of urinary pigments are presented. The justification of applying a correction to the values of NAG activity in human and rabbit urine (a model system for studying the toxic effects of cadmium) was discussed.",
publisher = "Wiley-Liss, Hoboken",
journal = "Journal of Clinical Laboratory Analysis",
title = "Influence of pigments and pH of urine on the determination of N-acetyl-beta-D-glucosaminidase activity with 2-methoxy-4-(2 '-nitrovinyl)-phenyl-N-acetyl-beta-D-glucosaminide",
volume = "19",
number = "6",
pages = "260-266",
doi = "10.1002/jcla.20088"
}

Aćimović, J. M., Jovanović, V. B.,& Mandić, L. M.. (2005). Influence of pigments and pH of urine on the determination of N-acetyl-beta-D-glucosaminidase activity with 2-methoxy-4-(2 '-nitrovinyl)-phenyl-N-acetyl-beta-D-glucosaminide. in Journal of Clinical Laboratory Analysis
Wiley-Liss, Hoboken., 19(6), 260-266.
https://doi.org/10.1002/jcla.20088

Aćimović JM, Jovanović VB, Mandić LM. Influence of pigments and pH of urine on the determination of N-acetyl-beta-D-glucosaminidase activity with 2-methoxy-4-(2 '-nitrovinyl)-phenyl-N-acetyl-beta-D-glucosaminide. in Journal of Clinical Laboratory Analysis. 2005;19(6):260-266.
doi:10.1002/jcla.20088 .

Aćimović, Jelena M., Jovanović, Vesna B., Mandić, Ljuba M., "Influence of pigments and pH of urine on the determination of N-acetyl-beta-D-glucosaminidase activity with 2-methoxy-4-(2 '-nitrovinyl)-phenyl-N-acetyl-beta-D-glucosaminide" in Journal of Clinical Laboratory Analysis, 19, no. 6 (2005):260-266,
https://doi.org/10.1002/jcla.20088 . .

TY  - JOUR
AU  - Mandić, Ljuba M.
AU  - Aćimović, Jelena M.
AU  - Jovanović, Vesna B.
PY  - 2005
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/692
AB  - Objectives: To have a reliable diagnostic test, the influence of urine pH on the determination of the total activity of N-acetyl-beta-D-glucosarninidase (NAG) and NAG isoenzyme activities was studied. Design and methods: After ultrafiltration and dialysis of the acidic and alkaline urines, the B, A, and A(2) forms of NAG were separated by ion-exchange chromatography on DEAE cellulose. Results: A significant decrease in the total activity of NAG in alkaline urines (pH around 8 or higher) was found, which makes this determination unreliable. Analysis of the isoenzymic profiles obtained for weakly acidic and alkaline urines (in the pH range from 5.5. to 10.0) showed that the percent fractions of the individual isoenzyme activities in the total NAG activity and their ratios changed only at pH values above 9.5. Conclusions: The determination of the denoted isoenzymes of urinary NAG after ultrafiltration, dialysis, and chromatographic separation on DEAE cellulose is reliable in a wide range of alkaline pH values Of urine. (c) 2005 The Canadian Society of Clinical Chemists. All rights reserved.
PB  - Pergamon-Elsevier Science Ltd, Oxford
T2  - Clinical Biochemistry
T1  - The possibility of determining N-acetyl-beta-D-glucosaminidase isoenzymes under alkaline conditions
VL  - 38
IS  - 4
SP  - 384
EP  - 389
DO  - 10.1016/j.clinbiochem.2005.01.002
ER  - 

@article{
author = "Mandić, Ljuba M. and Aćimović, Jelena M. and Jovanović, Vesna B.",
year = "2005",
abstract = "Objectives: To have a reliable diagnostic test, the influence of urine pH on the determination of the total activity of N-acetyl-beta-D-glucosarninidase (NAG) and NAG isoenzyme activities was studied. Design and methods: After ultrafiltration and dialysis of the acidic and alkaline urines, the B, A, and A(2) forms of NAG were separated by ion-exchange chromatography on DEAE cellulose. Results: A significant decrease in the total activity of NAG in alkaline urines (pH around 8 or higher) was found, which makes this determination unreliable. Analysis of the isoenzymic profiles obtained for weakly acidic and alkaline urines (in the pH range from 5.5. to 10.0) showed that the percent fractions of the individual isoenzyme activities in the total NAG activity and their ratios changed only at pH values above 9.5. Conclusions: The determination of the denoted isoenzymes of urinary NAG after ultrafiltration, dialysis, and chromatographic separation on DEAE cellulose is reliable in a wide range of alkaline pH values Of urine. (c) 2005 The Canadian Society of Clinical Chemists. All rights reserved.",
publisher = "Pergamon-Elsevier Science Ltd, Oxford",
journal = "Clinical Biochemistry",
title = "The possibility of determining N-acetyl-beta-D-glucosaminidase isoenzymes under alkaline conditions",
volume = "38",
number = "4",
pages = "384-389",
doi = "10.1016/j.clinbiochem.2005.01.002"
}

Mandić, L. M., Aćimović, J. M.,& Jovanović, V. B.. (2005). The possibility of determining N-acetyl-beta-D-glucosaminidase isoenzymes under alkaline conditions. in Clinical Biochemistry
Pergamon-Elsevier Science Ltd, Oxford., 38(4), 384-389.
https://doi.org/10.1016/j.clinbiochem.2005.01.002

Mandić LM, Aćimović JM, Jovanović VB. The possibility of determining N-acetyl-beta-D-glucosaminidase isoenzymes under alkaline conditions. in Clinical Biochemistry. 2005;38(4):384-389.
doi:10.1016/j.clinbiochem.2005.01.002 .

Mandić, Ljuba M., Aćimović, Jelena M., Jovanović, Vesna B., "The possibility of determining N-acetyl-beta-D-glucosaminidase isoenzymes under alkaline conditions" in Clinical Biochemistry, 38, no. 4 (2005):384-389,
https://doi.org/10.1016/j.clinbiochem.2005.01.002 . .