TY  - CONF
AU  - T. Milenković, Marina
AU  - Ušjak, Dušan
AU  - Tadić, Vanja
PY  - 2024
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2377
AB  - Antimicrobial resistance (AMR) has developed as
one of the top 10 global public health threats
facing humanity. As the nosocomial bacterial
strains are being increasingly resistant to most
clinically available antibiotics, there is a constant
need for exploration of new substances
that could kill them or inhibit their growth, or
alternatively inhibit some of their essential virulence
factors to counteract the lack of new antibacterials
and the rise of antibiotic resistance,
plants could represent a potential solution.
Plants produce a variety of bioactive secondary
metabolites that could be used to fuel the future
discovery pipeline. Aim of the present study was
to examine inhibitory activity of the supercritical
extract of J. communis L. green pseudofructus
(7SCO2) against the growth, biofilm production
and several virulence factors of significant nosocomial
bacterial pathogens. The extract was
obtained by fractional extraction with supercritical
CO2, and the qualitative and quantitative
analysis was performed using the GC-FID/MS
method. Clinical isolates of Pseudomonas aeruginosa,Acinetobacter baumannii, Staphylococcus
aureus (methicillin-sensitive-MSSA and methicillin-
resistant - MRSA), Enterococcus faecalis, and
Klebsiella pneumoniae, as well as their antibiotic
resistance profiles, were obtained from the Clinical
Hospital Centre “Dr Dragiša Mišović Dedinje”.
Minimum inhibitory concentrations (MICs) of
the 7SCO2 were determined by broth-microdilution
method. Examination of the anti-adhesive
effect of the extract was carried out using the
spectrophotometric method. The pyocyanin
production of Pseudomonas aeruginosa was determined
by the method described by Rampioni
et al. Most significant findings of this study
are potent antivirulence activity of the 7SCO2
against P. aeruginosa through the inhibition of
pyocyanin production. In addition, the biofilm
production of A. baumannii was inhibited by the
7SCO2 in concentration 50 μg/mL. Finally, notable
antivirulence activity of the 7SCO2 against
E. faecalis and S. aureus was detected, since it
significantly inhibited collagen and laminin adhesion
of these pathogens.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
T1  - HERBAL PRODUCTS AS AN ALTERNATIVE TO ANTIBIOTICS: APPLICATION POSSIBILITIES AND LIMITATIONS
EP  - 131
SP  - 131
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2377
ER  - 

@conference{
author = "T. Milenković, Marina and Ušjak, Dušan and Tadić, Vanja",
year = "2024",
abstract = "Antimicrobial resistance (AMR) has developed as
one of the top 10 global public health threats
facing humanity. As the nosocomial bacterial
strains are being increasingly resistant to most
clinically available antibiotics, there is a constant
need for exploration of new substances
that could kill them or inhibit their growth, or
alternatively inhibit some of their essential virulence
factors to counteract the lack of new antibacterials
and the rise of antibiotic resistance,
plants could represent a potential solution.
Plants produce a variety of bioactive secondary
metabolites that could be used to fuel the future
discovery pipeline. Aim of the present study was
to examine inhibitory activity of the supercritical
extract of J. communis L. green pseudofructus
(7SCO2) against the growth, biofilm production
and several virulence factors of significant nosocomial
bacterial pathogens. The extract was
obtained by fractional extraction with supercritical
CO2, and the qualitative and quantitative
analysis was performed using the GC-FID/MS
method. Clinical isolates of Pseudomonas aeruginosa,Acinetobacter baumannii, Staphylococcus
aureus (methicillin-sensitive-MSSA and methicillin-
resistant - MRSA), Enterococcus faecalis, and
Klebsiella pneumoniae, as well as their antibiotic
resistance profiles, were obtained from the Clinical
Hospital Centre “Dr Dragiša Mišović Dedinje”.
Minimum inhibitory concentrations (MICs) of
the 7SCO2 were determined by broth-microdilution
method. Examination of the anti-adhesive
effect of the extract was carried out using the
spectrophotometric method. The pyocyanin
production of Pseudomonas aeruginosa was determined
by the method described by Rampioni
et al. Most significant findings of this study
are potent antivirulence activity of the 7SCO2
against P. aeruginosa through the inhibition of
pyocyanin production. In addition, the biofilm
production of A. baumannii was inhibited by the
7SCO2 in concentration 50 μg/mL. Finally, notable
antivirulence activity of the 7SCO2 against
E. faecalis and S. aureus was detected, since it
significantly inhibited collagen and laminin adhesion
of these pathogens.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health",
title = "HERBAL PRODUCTS AS AN ALTERNATIVE TO ANTIBIOTICS: APPLICATION POSSIBILITIES AND LIMITATIONS",
pages = "131-131",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2377"
}

T. Milenković, M., Ušjak, D.,& Tadić, V.. (2024). HERBAL PRODUCTS AS AN ALTERNATIVE TO ANTIBIOTICS: APPLICATION POSSIBILITIES AND LIMITATIONS. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health
Serbian Society for Microbiology., 131-131.
https://hdl.handle.net/21.15107/rcub_imagine_2377

T. Milenković M, Ušjak D, Tadić V. HERBAL PRODUCTS AS AN ALTERNATIVE TO ANTIBIOTICS: APPLICATION POSSIBILITIES AND LIMITATIONS. in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health. 2024;:131-131.
https://hdl.handle.net/21.15107/rcub_imagine_2377 .

T. Milenković, Marina, Ušjak, Dušan, Tadić, Vanja, "HERBAL PRODUCTS AS AN ALTERNATIVE TO ANTIBIOTICS: APPLICATION POSSIBILITIES AND LIMITATIONS" in XIII Congress of microbiologists of Serbia: From biotechnology to human and planetary health (2024):131-131,
https://hdl.handle.net/21.15107/rcub_imagine_2377 .

TY  - JOUR
AU  - Filipić, Brankica
AU  - Ušjak, Dušan
AU  - Rambaher, Martina Hrast
AU  - Oljacic, Slavica
AU  - Milenković, Marina
PY  - 2024
UR  - https://www.frontiersin.org/articles/10.3389/fcimb.2024.1370062
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2333
AB  - Antimicrobial resistance is a global threat, leading to an alarming increase in the prevalence of bacterial infections that can no longer be treated with available antibiotics. The World Health Organization estimates that by 2050 up to 10 million deaths per year could be associated with antimicrobial resistance, which would equal the annual number of cancer deaths worldwide. To overcome this emerging crisis, novel anti-bacterial compounds are urgently needed. There are two possible approaches in the fight against bacterial infections: a) targeting structures within bacterial cells, similar to existing antibiotics; and/or b) targeting virulence factors rather than bacterial growth. Here, for the first time, we provide a comprehensive overview of the key steps in the evaluation of potential new anti-bacterial and/or anti-virulence compounds. The methods described in this review include: a) in silico methods for the evaluation of novel compounds; b) anti-bacterial assays (MIC, MBC, Time-kill); b) anti-virulence assays (anti-biofilm, anti-quorum sensing, anti-adhesion); and c) evaluation of safety aspects (cytotoxicity assay and Ames test). Overall, we provide a detailed description of the methods that are an essential tool for chemists, computational chemists, microbiologists, and toxicologists in the evaluation of potential novel antimicrobial compounds. These methods are cost-effective and have high predictive value. They are widely used in preclinical studies to identify new molecular candidates, for further investigation in animal and human trials.
PB  - Frontiers
T2  - Frontiers in Cellular and Infection Microbiology
T1  - Evaluation of novel compounds as anti-bacterial or anti-virulence agents
VL  - 14
DO  - 10.3389/fcimb.2024.1370062
ER  - 

@article{
author = "Filipić, Brankica and Ušjak, Dušan and Rambaher, Martina Hrast and Oljacic, Slavica and Milenković, Marina",
year = "2024",
abstract = "Antimicrobial resistance is a global threat, leading to an alarming increase in the prevalence of bacterial infections that can no longer be treated with available antibiotics. The World Health Organization estimates that by 2050 up to 10 million deaths per year could be associated with antimicrobial resistance, which would equal the annual number of cancer deaths worldwide. To overcome this emerging crisis, novel anti-bacterial compounds are urgently needed. There are two possible approaches in the fight against bacterial infections: a) targeting structures within bacterial cells, similar to existing antibiotics; and/or b) targeting virulence factors rather than bacterial growth. Here, for the first time, we provide a comprehensive overview of the key steps in the evaluation of potential new anti-bacterial and/or anti-virulence compounds. The methods described in this review include: a) in silico methods for the evaluation of novel compounds; b) anti-bacterial assays (MIC, MBC, Time-kill); b) anti-virulence assays (anti-biofilm, anti-quorum sensing, anti-adhesion); and c) evaluation of safety aspects (cytotoxicity assay and Ames test). Overall, we provide a detailed description of the methods that are an essential tool for chemists, computational chemists, microbiologists, and toxicologists in the evaluation of potential novel antimicrobial compounds. These methods are cost-effective and have high predictive value. They are widely used in preclinical studies to identify new molecular candidates, for further investigation in animal and human trials.",
publisher = "Frontiers",
journal = "Frontiers in Cellular and Infection Microbiology",
title = "Evaluation of novel compounds as anti-bacterial or anti-virulence agents",
volume = "14",
doi = "10.3389/fcimb.2024.1370062"
}

Filipić, B., Ušjak, D., Rambaher, M. H., Oljacic, S.,& Milenković, M.. (2024). Evaluation of novel compounds as anti-bacterial or anti-virulence agents. in Frontiers in Cellular and Infection Microbiology
Frontiers., 14.
https://doi.org/10.3389/fcimb.2024.1370062

Filipić B, Ušjak D, Rambaher MH, Oljacic S, Milenković M. Evaluation of novel compounds as anti-bacterial or anti-virulence agents. in Frontiers in Cellular and Infection Microbiology. 2024;14.
doi:10.3389/fcimb.2024.1370062 .

Filipić, Brankica, Ušjak, Dušan, Rambaher, Martina Hrast, Oljacic, Slavica, Milenković, Marina, "Evaluation of novel compounds as anti-bacterial or anti-virulence agents" in Frontiers in Cellular and Infection Microbiology, 14 (2024),
https://doi.org/10.3389/fcimb.2024.1370062 . .

TY  - JOUR
AU  - Mićović, Tijana
AU  - Ušjak, Dušan
AU  - Milenković, Marina
AU  - Samardžić, Stevan
AU  - Maksimović, Zoran
PY  - 2024
UR  - https://www.lekovitesirovine.rs/article/173
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2336
AB  - In this study, antimicrobial activity of essential oils extracted from the aerial flowering parts (herbs) of Hyssopus officinalis subsp. aristatus (Godr.) Nyman (Lamiaceae) collected from five different locations in Montenegro, or purchased in Serbia, were investigated. In addition, their antibacterial activity in combination with antibiotics was studied. The antimicrobial activity against selected standard bacterial and yeast strains was investigated using the broth microdilution method. Two standard antibiotics were used for comparison: the aminoglycoside antibiotic amikacin and the cephalosporin antibiotic ceftriaxone. The antimicrobial activity of the essential oil-amikacin combination was investigated using the checkerboard assay. The main components of the essential oils were 1,8-cineole, cis-pinocamphone, β-pinene and limonene in varying quantities. Most of the tested essential oils showed no significant antimicrobial activity. However, an essential oil rich in cis-pinocamphone showed moderate activity against both Staphylococcus aureus and Escherichia coli (MIC = 400 µg/mL). The overall effect of the essential oils and antibiotic combinations against E. coli or S. aureus ranged from additive (FICI = 0.625) to indifferent (FICI = 1.5), depending on the source of the essential oil.
PB  - Institute for Medicinal Plants Research "Dr. Josif Pancic"
T2  - Lekovite Sirovine
T1  - Antimicrobial activity of Hyssopus officinalis L. subsp. aristatus (Godr.) Nyman (Lamiaceae) essential oils from Montenegro and Serbia
EP  - 6
IS  - 1
SP  - 1
VL  - 43
DO  - 10.61652/leksir2343e173M
ER  - 

@article{
author = "Mićović, Tijana and Ušjak, Dušan and Milenković, Marina and Samardžić, Stevan and Maksimović, Zoran",
year = "2024",
abstract = "In this study, antimicrobial activity of essential oils extracted from the aerial flowering parts (herbs) of Hyssopus officinalis subsp. aristatus (Godr.) Nyman (Lamiaceae) collected from five different locations in Montenegro, or purchased in Serbia, were investigated. In addition, their antibacterial activity in combination with antibiotics was studied. The antimicrobial activity against selected standard bacterial and yeast strains was investigated using the broth microdilution method. Two standard antibiotics were used for comparison: the aminoglycoside antibiotic amikacin and the cephalosporin antibiotic ceftriaxone. The antimicrobial activity of the essential oil-amikacin combination was investigated using the checkerboard assay. The main components of the essential oils were 1,8-cineole, cis-pinocamphone, β-pinene and limonene in varying quantities. Most of the tested essential oils showed no significant antimicrobial activity. However, an essential oil rich in cis-pinocamphone showed moderate activity against both Staphylococcus aureus and Escherichia coli (MIC = 400 µg/mL). The overall effect of the essential oils and antibiotic combinations against E. coli or S. aureus ranged from additive (FICI = 0.625) to indifferent (FICI = 1.5), depending on the source of the essential oil.",
publisher = "Institute for Medicinal Plants Research "Dr. Josif Pancic"",
journal = "Lekovite Sirovine",
title = "Antimicrobial activity of Hyssopus officinalis L. subsp. aristatus (Godr.) Nyman (Lamiaceae) essential oils from Montenegro and Serbia",
pages = "6-1",
number = "1",
volume = "43",
doi = "10.61652/leksir2343e173M"
}

Mićović, T., Ušjak, D., Milenković, M., Samardžić, S.,& Maksimović, Z.. (2024). Antimicrobial activity of Hyssopus officinalis L. subsp. aristatus (Godr.) Nyman (Lamiaceae) essential oils from Montenegro and Serbia. in Lekovite Sirovine
Institute for Medicinal Plants Research "Dr. Josif Pancic"., 43(1), 1-6.
https://doi.org/10.61652/leksir2343e173M

Mićović T, Ušjak D, Milenković M, Samardžić S, Maksimović Z. Antimicrobial activity of Hyssopus officinalis L. subsp. aristatus (Godr.) Nyman (Lamiaceae) essential oils from Montenegro and Serbia. in Lekovite Sirovine. 2024;43(1):1-6.
doi:10.61652/leksir2343e173M .

Mićović, Tijana, Ušjak, Dušan, Milenković, Marina, Samardžić, Stevan, Maksimović, Zoran, "Antimicrobial activity of Hyssopus officinalis L. subsp. aristatus (Godr.) Nyman (Lamiaceae) essential oils from Montenegro and Serbia" in Lekovite Sirovine, 43, no. 1 (2024):1-6,
https://doi.org/10.61652/leksir2343e173M . .

TY  - CONF
AU  - Dunjić Manevski, Sofija
AU  - Cumbo, Marija
AU  - Gvozdenov, Maja
AU  - Tomić, Branko
AU  - Ušjak, Dušan
AU  - Đorđević
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2126
AB  - Introduction: The prothrombin Belgrade variant (c.1787G>A, p.Arg596Gln) is a rare mutation found in
Serbia, Japan, China, America, India and leads to antithrombin resistance. Prothrombin Belgrade mutation influencesthrombin-antithrombin interactions and leadsto impaired inactivation of mutated thrombin. Also, it affectssodium binding site in thrombin, which isimportant forswitching from fast thrombin
configuration (coagulant properties) to slow configuration (anticoagulant properties). It has only been
found in a heterozygous state, which could mean that homozygous carriers are incompatible with life.
By using prothrombin (FII) deficient plasma, we could reconstitute plasma of wild type, heterozygous and
homozygous carrier, which could give more insight into the mechanism of this mutation.
Methods: Recombinant wild type and mutated prothrombin were generated by transient transfection
in HEK293T cell line. Western blot analysis was performed to test the efficiency of transfection. Human
Prothrombin ELISA (Nordic BioSite, Sweden) was used in order to measure recombinant prothrombin
concentration. Overall Hemostasis Potential (OHP) assay was performed to assess recombinant protein
activity. Recombinant wild type and mutated prothrombin were added to FII deficient plasma (Siemens,
Germany) in order to create reconstituted plasma, in the final concentration of 0.1 mg/mL, as it is approximately the level of prothrombin in human plasma.
Results: Reconstituted plasma samples that correspond to non-carrier, heterozygous carrier, and homozygous mutation carrier plasma were reconstructed. Recombinant proteinstested by OHP assay were
functional.
Conclusion: Reconstituted plasma samples allow us to examine the mechanism of prothrombin Belgrade mutation in various assays and in homozygous form as well.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins
EP  - 71
SP  - 71
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2126
ER  - 

@conference{
author = "Dunjić Manevski, Sofija and Cumbo, Marija and Gvozdenov, Maja and Tomić, Branko and Ušjak, Dušan and Đorđević",
year = "2023",
abstract = "Introduction: The prothrombin Belgrade variant (c.1787G>A, p.Arg596Gln) is a rare mutation found in
Serbia, Japan, China, America, India and leads to antithrombin resistance. Prothrombin Belgrade mutation influencesthrombin-antithrombin interactions and leadsto impaired inactivation of mutated thrombin. Also, it affectssodium binding site in thrombin, which isimportant forswitching from fast thrombin
configuration (coagulant properties) to slow configuration (anticoagulant properties). It has only been
found in a heterozygous state, which could mean that homozygous carriers are incompatible with life.
By using prothrombin (FII) deficient plasma, we could reconstitute plasma of wild type, heterozygous and
homozygous carrier, which could give more insight into the mechanism of this mutation.
Methods: Recombinant wild type and mutated prothrombin were generated by transient transfection
in HEK293T cell line. Western blot analysis was performed to test the efficiency of transfection. Human
Prothrombin ELISA (Nordic BioSite, Sweden) was used in order to measure recombinant prothrombin
concentration. Overall Hemostasis Potential (OHP) assay was performed to assess recombinant protein
activity. Recombinant wild type and mutated prothrombin were added to FII deficient plasma (Siemens,
Germany) in order to create reconstituted plasma, in the final concentration of 0.1 mg/mL, as it is approximately the level of prothrombin in human plasma.
Results: Reconstituted plasma samples that correspond to non-carrier, heterozygous carrier, and homozygous mutation carrier plasma were reconstructed. Recombinant proteinstested by OHP assay were
functional.
Conclusion: Reconstituted plasma samples allow us to examine the mechanism of prothrombin Belgrade mutation in various assays and in homozygous form as well.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins",
pages = "71-71",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2126"
}

Dunjić Manevski, S., Cumbo, M., Gvozdenov, M., Tomić, B., Ušjak, D.,& Đorđević. (2023). Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 71-71.
https://hdl.handle.net/21.15107/rcub_imagine_2126

Dunjić Manevski S, Cumbo M, Gvozdenov M, Tomić B, Ušjak D, Đorđević. Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:71-71.
https://hdl.handle.net/21.15107/rcub_imagine_2126 .

Dunjić Manevski, Sofija, Cumbo, Marija, Gvozdenov, Maja, Tomić, Branko, Ušjak, Dušan, Đorđević, "Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):71-71,
https://hdl.handle.net/21.15107/rcub_imagine_2126 .

TY  - CONF
AU  - Divac Rankov, Aleksandra
AU  - Ušjak, Dušan
AU  - Mitić, Martina Mia
AU  - Kusić Tisma, Jelena
PY  - 2023
UR  - https://belbi.bg.ac.rs/
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1985
AB  - Cystic fibrosis (CF) is a monogenic autosomal recessive disease caused by mutations in
transmembrane conductance regulator (CFTR) gene. The golden standard for the diagnosis
of CF is sweat chloride testing (>60 mmol/L) together with the identification of two CFcausing
variants of CFTR gene. Nevertheless, about 0.01% of patients with elevated sweat
chloride and high clinical suspicion of CF do not carry any CF-causing variants.
Here we present analysis of whole exome sequencing (WES) results for two patients with
elevated sweat chloride levels and clinical presentation of CF in whom no CF-causing
mutations were detected after CFTR gene whole coding region sequencing, and large
insertion/deletion testing.
Genomic DNA was extracted from whole blood, subjected to library preparation using
DNA nanoball technology from BGI and sequenced on DNBSEQ-G400 (MGI). Produced
fastq files were mapped to hg38 reference genome using BWA/SAM tools. VCF files were
generated using GATK (BaseRecalibrator, HaplotypeCaller) and annotated with InterVar
and AnnoVar tools. Filtering of detected variants for disease relevance was done using
the following criteria: QC Filter, GnomAD Allele Frequency, Functional consequences and
phenotype-genotype relationship.
In both patients, similar number of variants predicted to impair protein function were
detected (27 and 25). In two genes (CACNA1H and MUC5B) missense type variants
were found in both patients and loss of function variants were found in 7 and 11 genes,
respectively. Functional assessment of selected variants is underway.
Bioinformatics analyses are a valuable tool enabling identification of underlining genetic
bases of disease phenotype, important in the context of optimal patient management and
targeted therapies.
PB  - Belgrade : Institute of molecular genetics and genetic engineering
C3  - 4th Belgrade Bioinformatics Conference
T1  - Decoding Cystic Fibrosis Phenotype
EP  - 44
SP  - 44
VL  - 4
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1985
ER  - 

@conference{
author = "Divac Rankov, Aleksandra and Ušjak, Dušan and Mitić, Martina Mia and Kusić Tisma, Jelena",
year = "2023",
abstract = "Cystic fibrosis (CF) is a monogenic autosomal recessive disease caused by mutations in
transmembrane conductance regulator (CFTR) gene. The golden standard for the diagnosis
of CF is sweat chloride testing (>60 mmol/L) together with the identification of two CFcausing
variants of CFTR gene. Nevertheless, about 0.01% of patients with elevated sweat
chloride and high clinical suspicion of CF do not carry any CF-causing variants.
Here we present analysis of whole exome sequencing (WES) results for two patients with
elevated sweat chloride levels and clinical presentation of CF in whom no CF-causing
mutations were detected after CFTR gene whole coding region sequencing, and large
insertion/deletion testing.
Genomic DNA was extracted from whole blood, subjected to library preparation using
DNA nanoball technology from BGI and sequenced on DNBSEQ-G400 (MGI). Produced
fastq files were mapped to hg38 reference genome using BWA/SAM tools. VCF files were
generated using GATK (BaseRecalibrator, HaplotypeCaller) and annotated with InterVar
and AnnoVar tools. Filtering of detected variants for disease relevance was done using
the following criteria: QC Filter, GnomAD Allele Frequency, Functional consequences and
phenotype-genotype relationship.
In both patients, similar number of variants predicted to impair protein function were
detected (27 and 25). In two genes (CACNA1H and MUC5B) missense type variants
were found in both patients and loss of function variants were found in 7 and 11 genes,
respectively. Functional assessment of selected variants is underway.
Bioinformatics analyses are a valuable tool enabling identification of underlining genetic
bases of disease phenotype, important in the context of optimal patient management and
targeted therapies.",
publisher = "Belgrade : Institute of molecular genetics and genetic engineering",
journal = "4th Belgrade Bioinformatics Conference",
title = "Decoding Cystic Fibrosis Phenotype",
pages = "44-44",
volume = "4",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1985"
}

Divac Rankov, A., Ušjak, D., Mitić, M. M.,& Kusić Tisma, J.. (2023). Decoding Cystic Fibrosis Phenotype. in 4th Belgrade Bioinformatics Conference
Belgrade : Institute of molecular genetics and genetic engineering., 4, 44-44.
https://hdl.handle.net/21.15107/rcub_imagine_1985

Divac Rankov A, Ušjak D, Mitić MM, Kusić Tisma J. Decoding Cystic Fibrosis Phenotype. in 4th Belgrade Bioinformatics Conference. 2023;4:44-44.
https://hdl.handle.net/21.15107/rcub_imagine_1985 .

Divac Rankov, Aleksandra, Ušjak, Dušan, Mitić, Martina Mia, Kusić Tisma, Jelena, "Decoding Cystic Fibrosis Phenotype" in 4th Belgrade Bioinformatics Conference, 4 (2023):44-44,
https://hdl.handle.net/21.15107/rcub_imagine_1985 .

TY  - CONF
AU  - Dunjić Manevski, Sofija
AU  - Cumbo, Marija
AU  - Gvozdenov, Maja
AU  - Tomić, Branko
AU  - Ušjak, Dušan
AU  - Đorđević, Valentina
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2123
AB  - Introduction: The prothrombin Belgrade variant (c.1787G>A, p.Arg596Gln) is a rare mutation found in
Serbia, Japan, China, America, India and leads to antithrombin resistance. Prothrombin Belgrade mutation influencesthrombin-antithrombin interactions and leadsto impaired inactivation of mutated thrombin. Also, it affectssodium binding site in thrombin, which isimportant forswitching from fast thrombin
configuration (coagulant properties) to slow configuration (anticoagulant properties). It has only been
found in a heterozygous state, which could mean that homozygous carriers are incompatible with life.
By using prothrombin (FII) deficient plasma, we could reconstitute plasma of wild type, heterozygous and
homozygous carrier, which could give more insight into the mechanism of this mutation.
Methods: Recombinant wild type and mutated prothrombin were generated by transient transfection
in HEK293T cell line. Western blot analysis was performed to test the efficiency of transfection. Human
Prothrombin ELISA (Nordic BioSite, Sweden) was used in order to measure recombinant prothrombin
concentration. Overall Hemostasis Potential (OHP) assay was performed to assess recombinant protein
activity. Recombinant wild type and mutated prothrombin were added to FII deficient plasma (Siemens,
Germany) in order to create reconstituted plasma, in the final concentration of 0.1 mg/mL, as it is approximately the level of prothrombin in human plasma.
Results: Reconstituted plasma samples that correspond to non-carrier, heterozygous carrier, and homozygous mutation carrier plasma were reconstructed. Recombinant proteinstested by OHP assay were
functional.
Conclusion: Reconstituted plasma samples allow us to examine the mechanism of prothrombin Belgrade mutation in various assays and in homozygous form as well.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins
EP  - 71
SP  - 71
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2123
ER  - 

@conference{
author = "Dunjić Manevski, Sofija and Cumbo, Marija and Gvozdenov, Maja and Tomić, Branko and Ušjak, Dušan and Đorđević, Valentina",
year = "2023",
abstract = "Introduction: The prothrombin Belgrade variant (c.1787G>A, p.Arg596Gln) is a rare mutation found in
Serbia, Japan, China, America, India and leads to antithrombin resistance. Prothrombin Belgrade mutation influencesthrombin-antithrombin interactions and leadsto impaired inactivation of mutated thrombin. Also, it affectssodium binding site in thrombin, which isimportant forswitching from fast thrombin
configuration (coagulant properties) to slow configuration (anticoagulant properties). It has only been
found in a heterozygous state, which could mean that homozygous carriers are incompatible with life.
By using prothrombin (FII) deficient plasma, we could reconstitute plasma of wild type, heterozygous and
homozygous carrier, which could give more insight into the mechanism of this mutation.
Methods: Recombinant wild type and mutated prothrombin were generated by transient transfection
in HEK293T cell line. Western blot analysis was performed to test the efficiency of transfection. Human
Prothrombin ELISA (Nordic BioSite, Sweden) was used in order to measure recombinant prothrombin
concentration. Overall Hemostasis Potential (OHP) assay was performed to assess recombinant protein
activity. Recombinant wild type and mutated prothrombin were added to FII deficient plasma (Siemens,
Germany) in order to create reconstituted plasma, in the final concentration of 0.1 mg/mL, as it is approximately the level of prothrombin in human plasma.
Results: Reconstituted plasma samples that correspond to non-carrier, heterozygous carrier, and homozygous mutation carrier plasma were reconstructed. Recombinant proteinstested by OHP assay were
functional.
Conclusion: Reconstituted plasma samples allow us to examine the mechanism of prothrombin Belgrade mutation in various assays and in homozygous form as well.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins",
pages = "71-71",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2123"
}

Dunjić Manevski, S., Cumbo, M., Gvozdenov, M., Tomić, B., Ušjak, D.,& Đorđević, V.. (2023). Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 71-71.
https://hdl.handle.net/21.15107/rcub_imagine_2123

Dunjić Manevski S, Cumbo M, Gvozdenov M, Tomić B, Ušjak D, Đorđević V. Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:71-71.
https://hdl.handle.net/21.15107/rcub_imagine_2123 .

Dunjić Manevski, Sofija, Cumbo, Marija, Gvozdenov, Maja, Tomić, Branko, Ušjak, Dušan, Đorđević, Valentina, "Reconstitution of non-carrier, heterozygous and homozygous prothrombin belgrade mutation carrier plasma using recombinant proteins" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):71-71,
https://hdl.handle.net/21.15107/rcub_imagine_2123 .

TY  - JOUR
AU  - Ušjak, Dušan
AU  - Novović, Katarina
AU  - Ivković, Branka
AU  - Tomić, Branko
AU  - Đorđević, Valentina
AU  - Milenković, Marina
PY  - 2023
UR  - https://doi.org/10.1080/08927014.2023.2215693
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1896
AB  - Biofilm production facilitates microbial colonization of wounds and catheters. Acinetobacter baumannii produces high levels of biofilm and causes difficult-to-treat nosocomial infections. Candida albicans is another strong biofilm producer which may facilitate A. baumannii adhesion by providing hyphae-mediated OmpA-binding sites. Here we tested the potential of 2′-hydroxychalcones to inhibit dual-species biofilm production of A. baumannii and Candida spp., and further predicted the mechanism of structure-related difference in activity. The results suggest that 2′-hydroxychalcones exhibit potent activity against Candida spp./A. baumannii dual-species biofilm production. Particularly active was trifluoromethyl-substituted derivative (p-CF3), which decreased C. albicans/A. baumannii biomass produced on vein-indwelling parts of the central venous catheterization set by up to 99%. Further, higher OmpA-binding affinity was also calculated for p-CF3, which together with demonstrated significant ompA-downregulating activity, suggests that superior antibiofilm activity of this chalcone against the tested dual-species community of A. baumannii is mediated through the OmpA.
T2  - Biofouling
T2  - Biofouling
T1  - Targeting outer membrane protein A (OmpA) – inhibitory effect of 2′-hydroxychalcone derivatives on Acinetobacter baumannii and Candida albicans dual-species biofilm formation
EP  - 11
SP  - 1
DO  - 10.1080/08927014.2023.2215693
ER  - 

@article{
author = "Ušjak, Dušan and Novović, Katarina and Ivković, Branka and Tomić, Branko and Đorđević, Valentina and Milenković, Marina",
year = "2023",
abstract = "Biofilm production facilitates microbial colonization of wounds and catheters. Acinetobacter baumannii produces high levels of biofilm and causes difficult-to-treat nosocomial infections. Candida albicans is another strong biofilm producer which may facilitate A. baumannii adhesion by providing hyphae-mediated OmpA-binding sites. Here we tested the potential of 2′-hydroxychalcones to inhibit dual-species biofilm production of A. baumannii and Candida spp., and further predicted the mechanism of structure-related difference in activity. The results suggest that 2′-hydroxychalcones exhibit potent activity against Candida spp./A. baumannii dual-species biofilm production. Particularly active was trifluoromethyl-substituted derivative (p-CF3), which decreased C. albicans/A. baumannii biomass produced on vein-indwelling parts of the central venous catheterization set by up to 99%. Further, higher OmpA-binding affinity was also calculated for p-CF3, which together with demonstrated significant ompA-downregulating activity, suggests that superior antibiofilm activity of this chalcone against the tested dual-species community of A. baumannii is mediated through the OmpA.",
journal = "Biofouling, Biofouling",
title = "Targeting outer membrane protein A (OmpA) – inhibitory effect of 2′-hydroxychalcone derivatives on Acinetobacter baumannii and Candida albicans dual-species biofilm formation",
pages = "11-1",
doi = "10.1080/08927014.2023.2215693"
}

Ušjak, D., Novović, K., Ivković, B., Tomić, B., Đorđević, V.,& Milenković, M.. (2023). Targeting outer membrane protein A (OmpA) – inhibitory effect of 2′-hydroxychalcone derivatives on Acinetobacter baumannii and Candida albicans dual-species biofilm formation. in Biofouling, 1-11.
https://doi.org/10.1080/08927014.2023.2215693

Ušjak D, Novović K, Ivković B, Tomić B, Đorđević V, Milenković M. Targeting outer membrane protein A (OmpA) – inhibitory effect of 2′-hydroxychalcone derivatives on Acinetobacter baumannii and Candida albicans dual-species biofilm formation. in Biofouling. 2023;:1-11.
doi:10.1080/08927014.2023.2215693 .

Ušjak, Dušan, Novović, Katarina, Ivković, Branka, Tomić, Branko, Đorđević, Valentina, Milenković, Marina, "Targeting outer membrane protein A (OmpA) – inhibitory effect of 2′-hydroxychalcone derivatives on Acinetobacter baumannii and Candida albicans dual-species biofilm formation" in Biofouling (2023):1-11,
https://doi.org/10.1080/08927014.2023.2215693 . .

TY  - CONF
AU  - Mitić, Martina Mia
AU  - Ušjak, Dušan
AU  - Milošević, Maja
AU  - Cumbo, Marija
AU  - Dunjić Manevski, Sofija
AU  - Tomić, Branko
AU  - Petrović, Ivana
AU  - Otašević, Petar
AU  - Micović, Slobodan
AU  - Bojić, Milovan
AU  - Đorđević, Valentina
PY  - 2023
UR  - https://belbi.bg.ac.rs/
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1999
AB  - Normal aortic valve consists of three cusps that develop in the embryonic stage. Unicuspid
aortic valve (UAV) is a rare congenital anomaly resulting in only one cusp with estimated
prevalence of 0.02% in general population. Aim of this study was to identify genetic
variants possibly associated with development of UAV. The study included 17 subjects,
namely 5 UAV patients and their healthy family members without UAV disorder. Total
DNA was isolated from venous blood samples and whole exomes sequencing (WES) was
performed using BGI’s WES protocol. Adapter-trimmed and quality-filtered reads (fastp)
were mapped to hg38 reference genome using BWA/SAMtools. VCF files were generated
using GATK (BaseRecalibrator, HaplotypeCaller) and annotated with InterVar and AnnoVar
tools. Rare heterozygous variants present in UAV patients were found in NOTCH1, TGFB2,
MYH6, EGFR, FBN2, C1R, ROBO4 and TBX5, genes associated with development of aortic
valves. Among these, most were missense mutations with damaging effects as predicted
using in silico tools (SIFT and/or Polyphen). Only mutation in MYH6 p.Ala1130Ser was
found in at least two different UAV patients. Also, rare homozygous missense mutation
p.Gly577Ser with high damaging potential was found in ADAMTS5 gene. Besides, highly
damaging heterozygous missense mutations were detected in gene interacting functional
partners (STRING) of genes associated with development of aortic valves: DVL1, THBS1,
NOTCH4, ADAMTS3, FBN1, NOTCH2, ADAM17, LRP5, WWTR1, C1S, ANKRD6 and TNNI1,
as well as homozygous in ACAN and KNG1. Taken together, malfunctions in ADAMTS5,
ACTA2, MYH6, FBN2, AXIN1, CELSR1 or TBX5 networks were found to be common in at
least two UAV patients, suggesting existence of genetic basis in UAV disorder, possibly as
a result of combined effects of multiple variants.
PB  - Belgrade : Institute of molecular genetics and genetic engineering
C3  - 4th Belgrade Bioinformatics Conference
T1  - Using whole exome sequencing to explore genetic basis of unicuspid aortic valve disease
EP  - 59
SP  - 59
VL  - 4
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1999
ER  - 

@conference{
author = "Mitić, Martina Mia and Ušjak, Dušan and Milošević, Maja and Cumbo, Marija and Dunjić Manevski, Sofija and Tomić, Branko and Petrović, Ivana and Otašević, Petar and Micović, Slobodan and Bojić, Milovan and Đorđević, Valentina",
year = "2023",
abstract = "Normal aortic valve consists of three cusps that develop in the embryonic stage. Unicuspid
aortic valve (UAV) is a rare congenital anomaly resulting in only one cusp with estimated
prevalence of 0.02% in general population. Aim of this study was to identify genetic
variants possibly associated with development of UAV. The study included 17 subjects,
namely 5 UAV patients and their healthy family members without UAV disorder. Total
DNA was isolated from venous blood samples and whole exomes sequencing (WES) was
performed using BGI’s WES protocol. Adapter-trimmed and quality-filtered reads (fastp)
were mapped to hg38 reference genome using BWA/SAMtools. VCF files were generated
using GATK (BaseRecalibrator, HaplotypeCaller) and annotated with InterVar and AnnoVar
tools. Rare heterozygous variants present in UAV patients were found in NOTCH1, TGFB2,
MYH6, EGFR, FBN2, C1R, ROBO4 and TBX5, genes associated with development of aortic
valves. Among these, most were missense mutations with damaging effects as predicted
using in silico tools (SIFT and/or Polyphen). Only mutation in MYH6 p.Ala1130Ser was
found in at least two different UAV patients. Also, rare homozygous missense mutation
p.Gly577Ser with high damaging potential was found in ADAMTS5 gene. Besides, highly
damaging heterozygous missense mutations were detected in gene interacting functional
partners (STRING) of genes associated with development of aortic valves: DVL1, THBS1,
NOTCH4, ADAMTS3, FBN1, NOTCH2, ADAM17, LRP5, WWTR1, C1S, ANKRD6 and TNNI1,
as well as homozygous in ACAN and KNG1. Taken together, malfunctions in ADAMTS5,
ACTA2, MYH6, FBN2, AXIN1, CELSR1 or TBX5 networks were found to be common in at
least two UAV patients, suggesting existence of genetic basis in UAV disorder, possibly as
a result of combined effects of multiple variants.",
publisher = "Belgrade : Institute of molecular genetics and genetic engineering",
journal = "4th Belgrade Bioinformatics Conference",
title = "Using whole exome sequencing to explore genetic basis of unicuspid aortic valve disease",
pages = "59-59",
volume = "4",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1999"
}

Mitić, M. M., Ušjak, D., Milošević, M., Cumbo, M., Dunjić Manevski, S., Tomić, B., Petrović, I., Otašević, P., Micović, S., Bojić, M.,& Đorđević, V.. (2023). Using whole exome sequencing to explore genetic basis of unicuspid aortic valve disease. in 4th Belgrade Bioinformatics Conference
Belgrade : Institute of molecular genetics and genetic engineering., 4, 59-59.
https://hdl.handle.net/21.15107/rcub_imagine_1999

Mitić MM, Ušjak D, Milošević M, Cumbo M, Dunjić Manevski S, Tomić B, Petrović I, Otašević P, Micović S, Bojić M, Đorđević V. Using whole exome sequencing to explore genetic basis of unicuspid aortic valve disease. in 4th Belgrade Bioinformatics Conference. 2023;4:59-59.
https://hdl.handle.net/21.15107/rcub_imagine_1999 .

Mitić, Martina Mia, Ušjak, Dušan, Milošević, Maja, Cumbo, Marija, Dunjić Manevski, Sofija, Tomić, Branko, Petrović, Ivana, Otašević, Petar, Micović, Slobodan, Bojić, Milovan, Đorđević, Valentina, "Using whole exome sequencing to explore genetic basis of unicuspid aortic valve disease" in 4th Belgrade Bioinformatics Conference, 4 (2023):59-59,
https://hdl.handle.net/21.15107/rcub_imagine_1999 .

TY  - CONF
AU  - Kusić Tisma, Jelena
AU  - Ušjak, Dušan
AU  - Mitić, Martina Mia
AU  - Divac Rankov, Aleksandra
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2178
AB  - Background: Cystic fibrosis (CF) is autosomal
recessive disorder characterized by chronic
respiratory problems and poor growth. CF is caused
by defect in transmembrane conductance regulator
(CFTR) protein. CF is diagnosed by sweat chloride
analysis (>60 mmol/L) with the identification of
two CF-causing variants of CFTR gene. With a
longstanding history of CFTR gene analysis, our
laboratory identified several patients with elevated
sweat chloride and clinical manifestations of CF in
whom no CF-causing mutations were detected after
sequencing of whole coding region and testing for
large insertion/deletion of CFTR gene. In order to
elucidate genetic background of conditions that
mimic CF we performed whole exome sequencing
(WES) in two such patients.
Methods: Library preparation was done using
DNA nanoball technology. Produced fastq files
were mapped to hg38. VCF files were generated
using GATK and annotated with InterVar and AnnoVar
tools. Variants filtering for disease relevance was done using the following criteria: QC, GnomAD
Allele Frequency, Functional consequences
and phenotype-genotype relationship.
Results: CACNA1H and MUC5B genes were
found to be impaired in both patients. Similar number
of variants predicted to impair protein function
were detected (27 and 25) in each patient. Loss of
function variants were found in 7 and 11 genes,
respectively.
Conclusion: Further assessment of selected
variants will clarify their functional effect and relevance
for the patient’s clinical phenotype. WES
analysis will help identify genetic aspects of disease
and assist in optimal patient management in about
0.01% of patients with elevated sweat chloride and
high clinical suspicion of CF that do not carry any
CF-causing variants.
PB  - Macedonian Academy of Sciences and Arts
C3  - International Journal of Medical Genetics
T1  - POTENTIAL NEW GENES INVOLVED IN CYSTIC FIBROSIS PHENOTYPE
EP  - 80
IS  - Supplement
SP  - 80
VL  - 26
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2178
ER  - 

@conference{
author = "Kusić Tisma, Jelena and Ušjak, Dušan and Mitić, Martina Mia and Divac Rankov, Aleksandra",
year = "2023",
abstract = "Background: Cystic fibrosis (CF) is autosomal
recessive disorder characterized by chronic
respiratory problems and poor growth. CF is caused
by defect in transmembrane conductance regulator
(CFTR) protein. CF is diagnosed by sweat chloride
analysis (>60 mmol/L) with the identification of
two CF-causing variants of CFTR gene. With a
longstanding history of CFTR gene analysis, our
laboratory identified several patients with elevated
sweat chloride and clinical manifestations of CF in
whom no CF-causing mutations were detected after
sequencing of whole coding region and testing for
large insertion/deletion of CFTR gene. In order to
elucidate genetic background of conditions that
mimic CF we performed whole exome sequencing
(WES) in two such patients.
Methods: Library preparation was done using
DNA nanoball technology. Produced fastq files
were mapped to hg38. VCF files were generated
using GATK and annotated with InterVar and AnnoVar
tools. Variants filtering for disease relevance was done using the following criteria: QC, GnomAD
Allele Frequency, Functional consequences
and phenotype-genotype relationship.
Results: CACNA1H and MUC5B genes were
found to be impaired in both patients. Similar number
of variants predicted to impair protein function
were detected (27 and 25) in each patient. Loss of
function variants were found in 7 and 11 genes,
respectively.
Conclusion: Further assessment of selected
variants will clarify their functional effect and relevance
for the patient’s clinical phenotype. WES
analysis will help identify genetic aspects of disease
and assist in optimal patient management in about
0.01% of patients with elevated sweat chloride and
high clinical suspicion of CF that do not carry any
CF-causing variants.",
publisher = "Macedonian Academy of Sciences and Arts",
journal = "International Journal of Medical Genetics",
title = "POTENTIAL NEW GENES INVOLVED IN CYSTIC FIBROSIS PHENOTYPE",
pages = "80-80",
number = "Supplement",
volume = "26",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2178"
}

Kusić Tisma, J., Ušjak, D., Mitić, M. M.,& Divac Rankov, A.. (2023). POTENTIAL NEW GENES INVOLVED IN CYSTIC FIBROSIS PHENOTYPE. in International Journal of Medical Genetics
Macedonian Academy of Sciences and Arts., 26(Supplement), 80-80.
https://hdl.handle.net/21.15107/rcub_imagine_2178

Kusić Tisma J, Ušjak D, Mitić MM, Divac Rankov A. POTENTIAL NEW GENES INVOLVED IN CYSTIC FIBROSIS PHENOTYPE. in International Journal of Medical Genetics. 2023;26(Supplement):80-80.
https://hdl.handle.net/21.15107/rcub_imagine_2178 .

Kusić Tisma, Jelena, Ušjak, Dušan, Mitić, Martina Mia, Divac Rankov, Aleksandra, "POTENTIAL NEW GENES INVOLVED IN CYSTIC FIBROSIS PHENOTYPE" in International Journal of Medical Genetics, 26, no. Supplement (2023):80-80,
https://hdl.handle.net/21.15107/rcub_imagine_2178 .

TY  - CONF
AU  - Cumbo, Marija
AU  - Tomić, Branko
AU  - Dunjić Manevski, Sofija
AU  - Gvozdenov, Maja
AU  - Ušjak, Dušan
AU  - Mitić, Martina Mia
AU  - Đorđević, Valentina
PY  - 2023
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/2135
AB  - Introduction: Thrombin, crucial member of the coagulation cascade, can influence growth and development of different types of cancer. Prothrombin, thrombin precursor, although predominantly secreted
from the liver into the bloodstream, can also be expressed in the cancer cells. According to latest data prothrombin can bind in vitro to transmembrane receptors, which have previously been shown to be up-regulated in cancers and activate migration and invasion. Despite the significant amount of data on the
effects of thrombin in cancer progression, there are little data of prothrombin´s effect. The aim of this
study was to further examine the effects of prothrombin and thrombin in cancer cell lines.
Methods: Colon cancer cell lines (Caco2, SW480, SW620, HT29 and HCT116) were treated with prothrombin, thrombin and direct thrombin inhibitor, dabigatran, for 24h and 48h. To assess the effects of
treatment on cell viability and proliferation MTT test was used, and wound healing assay was used for cell
migration potential.
Results: Detected effects of treatment with prothrombin, thrombin and dabigatran varied between cell
lines. Trend of lower cell viability, proliferation and migration was observed in cells treated with prothrombin in comparison to untreated controls.
Conclusion: Our resultsindicate that prothrombin, although considered an inactive zymogen, can exert
an effect on colon cancer cells proliferation and migration in vitro.
PB  - Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade
C3  - CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
T1  - Prothrombin influences proliferation and migration of colon cancer in vitro
EP  - 156
SP  - 156
UR  - https://hdl.handle.net/21.15107/rcub_imagine_2135
ER  - 

@conference{
author = "Cumbo, Marija and Tomić, Branko and Dunjić Manevski, Sofija and Gvozdenov, Maja and Ušjak, Dušan and Mitić, Martina Mia and Đorđević, Valentina",
year = "2023",
abstract = "Introduction: Thrombin, crucial member of the coagulation cascade, can influence growth and development of different types of cancer. Prothrombin, thrombin precursor, although predominantly secreted
from the liver into the bloodstream, can also be expressed in the cancer cells. According to latest data prothrombin can bind in vitro to transmembrane receptors, which have previously been shown to be up-regulated in cancers and activate migration and invasion. Despite the significant amount of data on the
effects of thrombin in cancer progression, there are little data of prothrombin´s effect. The aim of this
study was to further examine the effects of prothrombin and thrombin in cancer cell lines.
Methods: Colon cancer cell lines (Caco2, SW480, SW620, HT29 and HCT116) were treated with prothrombin, thrombin and direct thrombin inhibitor, dabigatran, for 24h and 48h. To assess the effects of
treatment on cell viability and proliferation MTT test was used, and wound healing assay was used for cell
migration potential.
Results: Detected effects of treatment with prothrombin, thrombin and dabigatran varied between cell
lines. Trend of lower cell viability, proliferation and migration was observed in cells treated with prothrombin in comparison to untreated controls.
Conclusion: Our resultsindicate that prothrombin, although considered an inactive zymogen, can exert
an effect on colon cancer cells proliferation and migration in vitro.",
publisher = "Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade",
journal = "CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia",
title = "Prothrombin influences proliferation and migration of colon cancer in vitro",
pages = "156-156",
url = "https://hdl.handle.net/21.15107/rcub_imagine_2135"
}

Cumbo, M., Tomić, B., Dunjić Manevski, S., Gvozdenov, M., Ušjak, D., Mitić, M. M.,& Đorđević, V.. (2023). Prothrombin influences proliferation and migration of colon cancer in vitro. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia
Institute of Molecular Genetics and Genetic Engineering (IMGGE), University of Belgrade., 156-156.
https://hdl.handle.net/21.15107/rcub_imagine_2135

Cumbo M, Tomić B, Dunjić Manevski S, Gvozdenov M, Ušjak D, Mitić MM, Đorđević V. Prothrombin influences proliferation and migration of colon cancer in vitro. in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia. 2023;:156-156.
https://hdl.handle.net/21.15107/rcub_imagine_2135 .

Cumbo, Marija, Tomić, Branko, Dunjić Manevski, Sofija, Gvozdenov, Maja, Ušjak, Dušan, Mitić, Martina Mia, Đorđević, Valentina, "Prothrombin influences proliferation and migration of colon cancer in vitro" in CoMBoS2 – the Second Congress of Molecular Biologists of Serbia, Abstract Book – Trends in Molecular Biology, Special issue 06-08 October 2023, Belgrade, Serbia (2023):156-156,
https://hdl.handle.net/21.15107/rcub_imagine_2135 .

TY  - DATA
AU  - Stevanović, Magdalena
AU  - Filipović, Nenad
AU  - Kuzmanović, Maja
AU  - Tomić, Nina
AU  - Ušjak, Dušan
AU  - Milenković, Marina
AU  - Zheng, Kai
AU  - Stampfl, Juergen
AU  - Boccaccin, Aldo
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1774
PB  - SAGE
T2  - Nanotechnology in Biomaterials
T1  - Supplementary data for the article: Stevanović, M. M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J., & Boccaccini, A. R. (2022). Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. Journal of Biomaterials Applications, 36(10), 1800–1811. [https://doi.org/10.1177/08853282211073731]
IS  - 10
VL  - 36
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1774
ER  - 

@misc{
author = "Stevanović, Magdalena and Filipović, Nenad and Kuzmanović, Maja and Tomić, Nina and Ušjak, Dušan and Milenković, Marina and Zheng, Kai and Stampfl, Juergen and Boccaccin, Aldo",
year = "2022",
publisher = "SAGE",
journal = "Nanotechnology in Biomaterials",
title = "Supplementary data for the article: Stevanović, M. M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J., & Boccaccini, A. R. (2022). Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. Journal of Biomaterials Applications, 36(10), 1800–1811. [https://doi.org/10.1177/08853282211073731]",
number = "10",
volume = "36",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1774"
}

Stevanović, M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J.,& Boccaccin, A.. (2022). Supplementary data for the article: Stevanović, M. M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J., & Boccaccini, A. R. (2022). Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. Journal of Biomaterials Applications, 36(10), 1800–1811. [https://doi.org/10.1177/08853282211073731]. in Nanotechnology in Biomaterials
SAGE., 36(10).
https://hdl.handle.net/21.15107/rcub_imagine_1774

Stevanović M, Filipović N, Kuzmanović M, Tomić N, Ušjak D, Milenković M, Zheng K, Stampfl J, Boccaccin A. Supplementary data for the article: Stevanović, M. M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J., & Boccaccini, A. R. (2022). Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. Journal of Biomaterials Applications, 36(10), 1800–1811. [https://doi.org/10.1177/08853282211073731]. in Nanotechnology in Biomaterials. 2022;36(10).
https://hdl.handle.net/21.15107/rcub_imagine_1774 .

Stevanović, Magdalena, Filipović, Nenad, Kuzmanović, Maja, Tomić, Nina, Ušjak, Dušan, Milenković, Marina, Zheng, Kai, Stampfl, Juergen, Boccaccin, Aldo, "Supplementary data for the article: Stevanović, M. M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J., & Boccaccini, A. R. (2022). Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. Journal of Biomaterials Applications, 36(10), 1800–1811. [https://doi.org/10.1177/08853282211073731]" in Nanotechnology in Biomaterials, 36, no. 10 (2022),
https://hdl.handle.net/21.15107/rcub_imagine_1774 .

TY  - THES
AU  - Ušjak, Dušan
PY  - 2022
UR  - https://eteze.bg.ac.rs/application/showtheses?thesesId=9037
UR  - https://fedorabg.bg.ac.rs/fedora/get/o:28480/bdef:Content/download
UR  - https://plus.cobiss.net/cobiss/sr/sr/bib/77543689
UR  - https://nardus.mpn.gov.rs/handle/123456789/21379
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1845
AB  - Acinetobacter baumannii je nozokomijalni, multirezistentni patogen, koga karakteriše sposobnost perzistencije na neživim površinama i mogućnost veoma brzog sticanja rezistencije na antibiotike. Danas su u svetu rasprostranjeni izrazito rezistentni sojevi protiv kojih u mnogim zdravstvenim ustanovama ne postoji efikasna terapija, a pronalazak alternativnih terapijskih pristupa je od izuzetne važnosti. Halkoni su jedinjenja sa potvrđenim antimikrobnim svojstvima i pokazanim različitim antivirulentnim aktivnostima. Ciljevi istraživanja ovog rada bili su određivanje profila rezistencije, ispitivanje mogućnosti kontaminacije antiseptika i ispitivanje produkcije biofilma identifikovanih kliničkih izolata A. baumannii, kao i sinteza derivata hidroksihalkona i ispitivanje njihovih antimikrobnih i antivirulentnih aktivnosti protiv ovih izolata. Osetljivost izolata na antibiotike ispitana je kombinacijom difuzionih, dilucionih i automatizovanih metoda, a identifikovani kolistin-rezistentni izolati dodatno su podvrgnuti sekvenciranju celog genoma (WGS) i genetički su okarakterisani. Takođe, mehanizmi rezistencije na kolistin ispitani su primenom komparativne analize genoma i Real-Time kvantitativne lančane reakcije polimeraze (RT-qPCR). Time-kill test je primenjen za ispitivanje perzistencije u antisepticima, a nivo produkcije biofilma ispitan je pod različitim uslovima kultivacije in vitro statičkom metodom uz bojenje safraninom. Derivati hidroksihalkona sintetisani su pomoću Claisen- Schmidt kondenzacije i njihove antimikrobne aktivnosti, samih i u kombinaciji sa antibioticima, ispitane su bujon-mikrodilucionom, Time-kill i Checkerboard analizom. Antivirulentne aktivnosti odabranih halkona procenjene su posredstvom uticaja na produkciju biofilma (monomikrobnog i polimikrobnog), vijabilnost biofilmskih ćelija, ekspresiju motiliteta, gensku ekspresiju faktora virulencije (OmpA, Bap i AbaI), adheziju A. baumannii na komponente ekstracelularnog matriksa (ECM), kao što su fibronektin i kolagen, i aktivnost sistema međućelijske komunikacije (Quorum- Sensing, QS). Klinički izolati A. baumannii gotovo uniformno bili su rezistentni na karbapeneme, a čak skoro 19% izolata bilo je rezistentno na kolistin, pripadajući tako ekstenzivno rezistentnom ili panrezistentnom fenotipu. Izolati su pokazali sposobnost kontaminacije antiseptika i produkcije velikih količina biofilma. Nutritivni sastav hranljivih medijuma značajno je uticao na nivo produkcije biofilma, dok se visok nivo produkcije održao pri širokom opsegu različitih temperatura inkubacije i u prisustvu subinhibitornih koncentracija antibiotika. Sintetisani halkoni ispoljili su umerenu antimikrobnu aktivnost, pri čemu su metoksi-supstituisani derivati u proseku najjače inhibirali rast. Takođe, zabeleženo je nekoliko sinergističkih interakcija halkona sa meropenemom, a inhibicija efluksnih pumpi predložena je kao potencijalni mehanizam. Halkoni su pokazali sposobnost značajne inhibicije motiliteta i produkcije biofilma, a metoksi-supstituisani derivat (o- OCH3) ispoljio je značajnu antivirulentnu aktivnost posredstvom nishodne regulacije ekspresije ompA, bap i abaI gena i inhibicije adhezije na komponente ECM. Na osnovu ovih rezultata, o- OCH3 halkon je identifikovan kao potentni antivirulentni agens protiv A. baumannii.
AB  - Acinetobacter baumannii is a nosocomial, multiresistant pathogen, able to persist on abiotic surfaces and to rapidly acquire antibiotic resistance. Nowadays, highly resistant strains are widely disseminated throughout the world, and the discovery of alternative therapeutic strategies is of utter importance. Chalcones are compounds whose antimicrobial properties are well-known and for which different antivirulence activities have been demonstrated. The aims of this research were to determine resistance profiles, to evaluate the possibility of antiseptic contamination, and to analyze the biofilm production of identified A. baumannii clinical isolates, as well as to synthesize hydroxychalcone derivatives and to investigate their antimicrobial and antivirulence activities against these isolates. Antibiotic susceptibility of the isolates was tested by combination of diffusion, dilution, and automated methods, and additionally, identified colistin-resistant isolates were subjected to whole genome sequencing (WGS) and were genetically characterized. Also, colistin resistance mechanisms were explored by using comparative genome analysis and Real-Time quantitative polymerase chain reaction (RT-qPCR). Time-kill test was used for the measurement of bacterial survival in antiseptics, whereas the level of biofilm production under different cultivation conditions was quantified by in vitro static method using safranin stain. Hydroxychalcone derivatives were synthesized by Claisen-Schmidt condensation, and their antimicrobial activities, alone and in combination with antibiotics, were investigated using broth-microdilution, Time-kill, and Checkerboard analyses. Antivirulence activities of selected chalcones were evaluated based on the impact on biofilm production (monomicrobial and polymicrobial), biofilm cell viability, motility, virulence factors (OmpA, Bap, and AbaI) gene expression, fibronectin- and collagen- mediated adhesion, and quorum-sensing (QS) activity. A. baumannii clinical isolates expressed extensive drug-resistant or pan-drug resistant phenotypes, being nearly uniformly resistant to carbapenems. Almost 19% of isolates were resistant to colistin as well. The isolates proved to be able to contaminate the antiseptic solutions and to produce large quantities of biofilms. Nutritional composition of growth media significantly affected the level of biofilm production. In contrast, wide range of different incubation temperatures and the presence of antibiotics at subinhibitory concentrations had little effect, and the bacteria managed to maintain high level of biofilm production. Moderate antimicrobial activity was displayed by synthesized chalcones, among which methoxy-substituted derivatives achieved greatest growth inhibition in average. Also, synergistic activity of chalcones and meropenem was present in several cases, for which efflux pump inhibition was proposed as the potential mechanism. The chalcones significantly inhibited motility and biofilm production, whereas methoxy-substituted derivative (o- OCH3) also displayed significant antivirulence activity, by downregulating the ompA, bap, and abaI gene expression and by inhibiting fibronectin- and collagen-mediated adhesion. It can be concluded that o-OCH3 has been identified as a potent antivirulence agent against A. baumannii.
PB  - Beograd : Univerzitet u Beogradu, Farmaceutski fakultet
T1  - Uticaj novosintetisanih derivata halkona na rast, produkciju biofilma i faktore virulencije multirezistentnih sojeva Acinetobacter baumannii
T1  - Influence of newly-synthesized chalcone derivatives on growth, biofilm production, and virulence factors expression of multiresistant Acinetobacter baumannii strains
UR  - https://hdl.handle.net/21.15107/rcub_imagine_1845
ER  - 

@phdthesis{
author = "Ušjak, Dušan",
year = "2022",
abstract = "Acinetobacter baumannii je nozokomijalni, multirezistentni patogen, koga karakteriše sposobnost perzistencije na neživim površinama i mogućnost veoma brzog sticanja rezistencije na antibiotike. Danas su u svetu rasprostranjeni izrazito rezistentni sojevi protiv kojih u mnogim zdravstvenim ustanovama ne postoji efikasna terapija, a pronalazak alternativnih terapijskih pristupa je od izuzetne važnosti. Halkoni su jedinjenja sa potvrđenim antimikrobnim svojstvima i pokazanim različitim antivirulentnim aktivnostima. Ciljevi istraživanja ovog rada bili su određivanje profila rezistencije, ispitivanje mogućnosti kontaminacije antiseptika i ispitivanje produkcije biofilma identifikovanih kliničkih izolata A. baumannii, kao i sinteza derivata hidroksihalkona i ispitivanje njihovih antimikrobnih i antivirulentnih aktivnosti protiv ovih izolata. Osetljivost izolata na antibiotike ispitana je kombinacijom difuzionih, dilucionih i automatizovanih metoda, a identifikovani kolistin-rezistentni izolati dodatno su podvrgnuti sekvenciranju celog genoma (WGS) i genetički su okarakterisani. Takođe, mehanizmi rezistencije na kolistin ispitani su primenom komparativne analize genoma i Real-Time kvantitativne lančane reakcije polimeraze (RT-qPCR). Time-kill test je primenjen za ispitivanje perzistencije u antisepticima, a nivo produkcije biofilma ispitan je pod različitim uslovima kultivacije in vitro statičkom metodom uz bojenje safraninom. Derivati hidroksihalkona sintetisani su pomoću Claisen- Schmidt kondenzacije i njihove antimikrobne aktivnosti, samih i u kombinaciji sa antibioticima, ispitane su bujon-mikrodilucionom, Time-kill i Checkerboard analizom. Antivirulentne aktivnosti odabranih halkona procenjene su posredstvom uticaja na produkciju biofilma (monomikrobnog i polimikrobnog), vijabilnost biofilmskih ćelija, ekspresiju motiliteta, gensku ekspresiju faktora virulencije (OmpA, Bap i AbaI), adheziju A. baumannii na komponente ekstracelularnog matriksa (ECM), kao što su fibronektin i kolagen, i aktivnost sistema međućelijske komunikacije (Quorum- Sensing, QS). Klinički izolati A. baumannii gotovo uniformno bili su rezistentni na karbapeneme, a čak skoro 19% izolata bilo je rezistentno na kolistin, pripadajući tako ekstenzivno rezistentnom ili panrezistentnom fenotipu. Izolati su pokazali sposobnost kontaminacije antiseptika i produkcije velikih količina biofilma. Nutritivni sastav hranljivih medijuma značajno je uticao na nivo produkcije biofilma, dok se visok nivo produkcije održao pri širokom opsegu različitih temperatura inkubacije i u prisustvu subinhibitornih koncentracija antibiotika. Sintetisani halkoni ispoljili su umerenu antimikrobnu aktivnost, pri čemu su metoksi-supstituisani derivati u proseku najjače inhibirali rast. Takođe, zabeleženo je nekoliko sinergističkih interakcija halkona sa meropenemom, a inhibicija efluksnih pumpi predložena je kao potencijalni mehanizam. Halkoni su pokazali sposobnost značajne inhibicije motiliteta i produkcije biofilma, a metoksi-supstituisani derivat (o- OCH3) ispoljio je značajnu antivirulentnu aktivnost posredstvom nishodne regulacije ekspresije ompA, bap i abaI gena i inhibicije adhezije na komponente ECM. Na osnovu ovih rezultata, o- OCH3 halkon je identifikovan kao potentni antivirulentni agens protiv A. baumannii., Acinetobacter baumannii is a nosocomial, multiresistant pathogen, able to persist on abiotic surfaces and to rapidly acquire antibiotic resistance. Nowadays, highly resistant strains are widely disseminated throughout the world, and the discovery of alternative therapeutic strategies is of utter importance. Chalcones are compounds whose antimicrobial properties are well-known and for which different antivirulence activities have been demonstrated. The aims of this research were to determine resistance profiles, to evaluate the possibility of antiseptic contamination, and to analyze the biofilm production of identified A. baumannii clinical isolates, as well as to synthesize hydroxychalcone derivatives and to investigate their antimicrobial and antivirulence activities against these isolates. Antibiotic susceptibility of the isolates was tested by combination of diffusion, dilution, and automated methods, and additionally, identified colistin-resistant isolates were subjected to whole genome sequencing (WGS) and were genetically characterized. Also, colistin resistance mechanisms were explored by using comparative genome analysis and Real-Time quantitative polymerase chain reaction (RT-qPCR). Time-kill test was used for the measurement of bacterial survival in antiseptics, whereas the level of biofilm production under different cultivation conditions was quantified by in vitro static method using safranin stain. Hydroxychalcone derivatives were synthesized by Claisen-Schmidt condensation, and their antimicrobial activities, alone and in combination with antibiotics, were investigated using broth-microdilution, Time-kill, and Checkerboard analyses. Antivirulence activities of selected chalcones were evaluated based on the impact on biofilm production (monomicrobial and polymicrobial), biofilm cell viability, motility, virulence factors (OmpA, Bap, and AbaI) gene expression, fibronectin- and collagen- mediated adhesion, and quorum-sensing (QS) activity. A. baumannii clinical isolates expressed extensive drug-resistant or pan-drug resistant phenotypes, being nearly uniformly resistant to carbapenems. Almost 19% of isolates were resistant to colistin as well. The isolates proved to be able to contaminate the antiseptic solutions and to produce large quantities of biofilms. Nutritional composition of growth media significantly affected the level of biofilm production. In contrast, wide range of different incubation temperatures and the presence of antibiotics at subinhibitory concentrations had little effect, and the bacteria managed to maintain high level of biofilm production. Moderate antimicrobial activity was displayed by synthesized chalcones, among which methoxy-substituted derivatives achieved greatest growth inhibition in average. Also, synergistic activity of chalcones and meropenem was present in several cases, for which efflux pump inhibition was proposed as the potential mechanism. The chalcones significantly inhibited motility and biofilm production, whereas methoxy-substituted derivative (o- OCH3) also displayed significant antivirulence activity, by downregulating the ompA, bap, and abaI gene expression and by inhibiting fibronectin- and collagen-mediated adhesion. It can be concluded that o-OCH3 has been identified as a potent antivirulence agent against A. baumannii.",
publisher = "Beograd : Univerzitet u Beogradu, Farmaceutski fakultet",
title = "Uticaj novosintetisanih derivata halkona na rast, produkciju biofilma i faktore virulencije multirezistentnih sojeva Acinetobacter baumannii, Influence of newly-synthesized chalcone derivatives on growth, biofilm production, and virulence factors expression of multiresistant Acinetobacter baumannii strains",
url = "https://hdl.handle.net/21.15107/rcub_imagine_1845"
}

Ušjak, D.. (2022). Uticaj novosintetisanih derivata halkona na rast, produkciju biofilma i faktore virulencije multirezistentnih sojeva Acinetobacter baumannii. 
Beograd : Univerzitet u Beogradu, Farmaceutski fakultet..
https://hdl.handle.net/21.15107/rcub_imagine_1845

Ušjak D. Uticaj novosintetisanih derivata halkona na rast, produkciju biofilma i faktore virulencije multirezistentnih sojeva Acinetobacter baumannii. 2022;.
https://hdl.handle.net/21.15107/rcub_imagine_1845 .

Ušjak, Dušan, "Uticaj novosintetisanih derivata halkona na rast, produkciju biofilma i faktore virulencije multirezistentnih sojeva Acinetobacter baumannii" (2022),
https://hdl.handle.net/21.15107/rcub_imagine_1845 .

TY  - JOUR
AU  - Stevanović, Magdalena
AU  - Filipović, Nenad
AU  - Kuzmanović, Maja
AU  - Tomić, Nina
AU  - Ušjak, Dušan
AU  - Milenković, Marina
AU  - Zheng, Kai
AU  - Stampfl, Juergen
AU  - Boccaccin, Aldo
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1773
AB  - Multidrug-resistant bacterial strains represent an emerging global health threat and a great obstacle for bone tissue
engineering. One of the major components of the extracellular matrix of the bone is a collagen protein, while selenium is an
element that has antimicrobial potential, and is also important for bone metabolism and bone health. Here we represent the
incorporation of selenium nanoparticles (SeNPs) synthesized by the green chemical reduction method into collagen gels to
produce a composite material, collagen/SeNPs, with antimicrobial properties. The samples were comprehensively
characterized by zeta potential measurements, dynamic light scattering inductively coupled plasma-mass spectrometry
(ICP-MS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), optical microscopy, field-emission
scanning electron microscopy (FE-SEM), and differential scanning calorimetry The cytotoxicity of the SeNPS, as well as
collagen/SeNPs, was tested on the MRC-5 cells. It was revealed that collagen/SeNPS expressed a lower cytotoxic effect.
Collagen/SeNPs showed significant antibacterial activity against all tested Gram-positive strains, the major causative agents
of orthopedic infections as well as Candida albicans. Furthermore, three-dimensional β-tricalcium phosphate (3D-TCP)
scaffolds were fabricated by a well-established 3D printing (lithography) method, and afterward preliminary coated by
newly-synthesized SeNPs or collagen/SeNPs. In addition, uncoated 3D-TCP scaffolds as well as coated by collagen/SeNPs
were subjected to biofilm formation. The production of Staphylococcus aureus biofilm on coated scaffolds by collagen/SeNPs
was significantly reduced compared to the uncoated ones.
PB  - SAGE
T2  - Nanotechnology in Biomaterials
T1  - Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles
IS  - 10
VL  - 36
DO  - 10.1177/08853282211073731
ER  - 

@article{
author = "Stevanović, Magdalena and Filipović, Nenad and Kuzmanović, Maja and Tomić, Nina and Ušjak, Dušan and Milenković, Marina and Zheng, Kai and Stampfl, Juergen and Boccaccin, Aldo",
year = "2022",
abstract = "Multidrug-resistant bacterial strains represent an emerging global health threat and a great obstacle for bone tissue
engineering. One of the major components of the extracellular matrix of the bone is a collagen protein, while selenium is an
element that has antimicrobial potential, and is also important for bone metabolism and bone health. Here we represent the
incorporation of selenium nanoparticles (SeNPs) synthesized by the green chemical reduction method into collagen gels to
produce a composite material, collagen/SeNPs, with antimicrobial properties. The samples were comprehensively
characterized by zeta potential measurements, dynamic light scattering inductively coupled plasma-mass spectrometry
(ICP-MS), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), optical microscopy, field-emission
scanning electron microscopy (FE-SEM), and differential scanning calorimetry The cytotoxicity of the SeNPS, as well as
collagen/SeNPs, was tested on the MRC-5 cells. It was revealed that collagen/SeNPS expressed a lower cytotoxic effect.
Collagen/SeNPs showed significant antibacterial activity against all tested Gram-positive strains, the major causative agents
of orthopedic infections as well as Candida albicans. Furthermore, three-dimensional β-tricalcium phosphate (3D-TCP)
scaffolds were fabricated by a well-established 3D printing (lithography) method, and afterward preliminary coated by
newly-synthesized SeNPs or collagen/SeNPs. In addition, uncoated 3D-TCP scaffolds as well as coated by collagen/SeNPs
were subjected to biofilm formation. The production of Staphylococcus aureus biofilm on coated scaffolds by collagen/SeNPs
was significantly reduced compared to the uncoated ones.",
publisher = "SAGE",
journal = "Nanotechnology in Biomaterials",
title = "Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles",
number = "10",
volume = "36",
doi = "10.1177/08853282211073731"
}

Stevanović, M., Filipović, N., Kuzmanović, M., Tomić, N., Ušjak, D., Milenković, M., Zheng, K., Stampfl, J.,& Boccaccin, A.. (2022). Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. in Nanotechnology in Biomaterials
SAGE., 36(10).
https://doi.org/10.1177/08853282211073731

Stevanović M, Filipović N, Kuzmanović M, Tomić N, Ušjak D, Milenković M, Zheng K, Stampfl J, Boccaccin A. Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles. in Nanotechnology in Biomaterials. 2022;36(10).
doi:10.1177/08853282211073731 .

Stevanović, Magdalena, Filipović, Nenad, Kuzmanović, Maja, Tomić, Nina, Ušjak, Dušan, Milenković, Marina, Zheng, Kai, Stampfl, Juergen, Boccaccin, Aldo, "Synthesis and characterization of a collagen-based composite material containing selenium nanoparticles" in Nanotechnology in Biomaterials, 36, no. 10 (2022),
https://doi.org/10.1177/08853282211073731 . .

TY  - JOUR
AU  - Ušjak, Dušan
AU  - Novović, Katarina
AU  - Filipić, Brankica
AU  - Kojić, Milan
AU  - Filipović, Nenad
AU  - Stevanović, Magdalena
AU  - Milenković, Marina T.
PY  - 2022
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1600
AB  - Aims To investigate the synergistic activity of colistin and selenium nanoparticles (SeNPs) against pandrug-resistant (PDR) Ac. baumannii. Methods and Results Chequerboard and time-kill assays were employed to explore the potential synergistic interactions between colistin and SeNPs against Ac. baumannii isolates (8), previously determined as colistin-resistant (MIC range 16-256 mu g ml(-1)). Also, whole-genome sequencing (WGS) and gene expression analyses were used to elucidate the mechanisms of colistin resistance. Exceptionally strong synergistic activity (FICI range 0.004-0.035) of colistin and SeNPs against colistin-resistant isolates was revealed. Colistin (0.5 or 1 mu g ml(-1)) used in combination with SeNPs (0.5 mu g ml(-1)) was able to reduce initial inoculum during the first 4 h of incubation, in contrast to colistin (0.5, 1 or 2 mu g ml(-1)) alone. Conclusions These findings propose colistin/SeNPs combination as a new option to fight PDR Ac. baumannii, the therapeutic possibilities of which should be proved in future in vivo studies. Significance and Impact of Study Here we present the first evidence of synergy between colistin and selenium compounds against bacteria in general. Also, WGS and gene expression analyses provide some new insights into Ac. baumannii colistin resistance mechanisms.
PB  - Wiley, Hoboken
T2  - Journal of Applied Microbiology
T1  - In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles
EP  - 1206
IS  - 3
SP  - 1197
VL  - 133
DO  - 10.1111/jam.15638
ER  - 

@article{
author = "Ušjak, Dušan and Novović, Katarina and Filipić, Brankica and Kojić, Milan and Filipović, Nenad and Stevanović, Magdalena and Milenković, Marina T.",
year = "2022",
abstract = "Aims To investigate the synergistic activity of colistin and selenium nanoparticles (SeNPs) against pandrug-resistant (PDR) Ac. baumannii. Methods and Results Chequerboard and time-kill assays were employed to explore the potential synergistic interactions between colistin and SeNPs against Ac. baumannii isolates (8), previously determined as colistin-resistant (MIC range 16-256 mu g ml(-1)). Also, whole-genome sequencing (WGS) and gene expression analyses were used to elucidate the mechanisms of colistin resistance. Exceptionally strong synergistic activity (FICI range 0.004-0.035) of colistin and SeNPs against colistin-resistant isolates was revealed. Colistin (0.5 or 1 mu g ml(-1)) used in combination with SeNPs (0.5 mu g ml(-1)) was able to reduce initial inoculum during the first 4 h of incubation, in contrast to colistin (0.5, 1 or 2 mu g ml(-1)) alone. Conclusions These findings propose colistin/SeNPs combination as a new option to fight PDR Ac. baumannii, the therapeutic possibilities of which should be proved in future in vivo studies. Significance and Impact of Study Here we present the first evidence of synergy between colistin and selenium compounds against bacteria in general. Also, WGS and gene expression analyses provide some new insights into Ac. baumannii colistin resistance mechanisms.",
publisher = "Wiley, Hoboken",
journal = "Journal of Applied Microbiology",
title = "In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles",
pages = "1206-1197",
number = "3",
volume = "133",
doi = "10.1111/jam.15638"
}

Ušjak, D., Novović, K., Filipić, B., Kojić, M., Filipović, N., Stevanović, M.,& Milenković, M. T.. (2022). In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles. in Journal of Applied Microbiology
Wiley, Hoboken., 133(3), 1197-1206.
https://doi.org/10.1111/jam.15638

Ušjak D, Novović K, Filipić B, Kojić M, Filipović N, Stevanović M, Milenković MT. In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles. in Journal of Applied Microbiology. 2022;133(3):1197-1206.
doi:10.1111/jam.15638 .

Ušjak, Dušan, Novović, Katarina, Filipić, Brankica, Kojić, Milan, Filipović, Nenad, Stevanović, Magdalena, Milenković, Marina T., "In vitro colistin susceptibility of pandrug-resistant Ac. baumannii is restored in the presence of selenium nanoparticles" in Journal of Applied Microbiology, 133, no. 3 (2022):1197-1206,
https://doi.org/10.1111/jam.15638 . .

TY  - JOUR
AU  - Filipović, Nenad
AU  - Ušjak, Dušan
AU  - Milenković, Marina T.
AU  - Zheng, Kai
AU  - Liverani, Liliana
AU  - Boccaccini, Aldo R.
AU  - Stevanović, Magdalena M.
PY  - 2021
UR  - https://www.frontiersin.org/articles/10.3389/fbioe.2020.624621
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1772
AB  - Although selenium nanoparticles (SeNPs) have gained attention in the scientific community mostly through investigation of their anticancer activity, a great potential of this nanomaterial was recognized recently regarding its antimicrobial activity. The particle form, size, and surface chemistry have been recognized as crucial parameters determining the interaction of nanomaterials with biological entities. Furthermore, considering a narrow boundary between beneficial and toxic effects for selenium per se, it is clear that investigations of biomedical applications of SeNPs are very demanding and must be done with great precautions. The goal of this work is to evaluate the effects of SeNPs surface chemistry and structure on antimicrobial activity against several common bacterial strains, including Staphylococcus aureus (ATCC 6538), Enterococcus faecalis (ATCC 29212), Bacillus subtilis (ATCC 6633), and Kocuria rhizophila (ATCC 9341), as well as Escherichia coli (ATCC 8739), Salmonella Abony (NCTC 6017), Klebsiella pneumoniae (NCIMB 9111) and Pseudomonas aeruginosa (ATCC 9027), and the standard yeast strain Candida albicans (ATCC 10231). Three types of SeNPs were synthesized by chemical reduction approach using different stabilizers and reducing agents: (i) bovine serum albumin (BSA) + ascorbic acid, (ii) chitosan + ascorbic acid, and (iii) with glucose. A thorough physicochemical characterization of the obtained SeNPs was performed to determine the effects of varying synthesis parameters on their morphology, size, structure, and surface chemistry. All SeNPs were amorphous, with spherical morphology and size in the range 70–300 nm. However, the SeNPs obtained under different synthesis conditions, i.e. by using different stabilizers as well as reducing agents, exhibited different antimicrobial activity as well as cytotoxicity which are crucial for their applications. In this paper, the antimicrobial screening of the selected systems is presented, which was determined by the broth microdilution method, and inhibitory influence on the production of monomicrobial and dual-species biofilm was evaluated. The potential mechanism of action of different systems is proposed. Additionally, the cytotoxicity of SeNPs was examined on the MRC-5 cell line, in the same concentration interval as for antimicrobial testing. It was shown that formulation SeNPs-BSA expressed a significantly lower cytotoxic effect than the other two formulations.
PB  - Frontiers
T2  - Frontiers in Bioengineering and Biotechnology
T2  - Frontiers in Bioengineering and Biotechnology
T1  - Comparative Study of the Antimicrobial Activity of Selenium Nanoparticles With Different Surface Chemistry and Structure
VL  - 8
DO  - 10.3389/fbioe.2020.624621
ER  - 

@article{
author = "Filipović, Nenad and Ušjak, Dušan and Milenković, Marina T. and Zheng, Kai and Liverani, Liliana and Boccaccini, Aldo R. and Stevanović, Magdalena M.",
year = "2021",
abstract = "Although selenium nanoparticles (SeNPs) have gained attention in the scientific community mostly through investigation of their anticancer activity, a great potential of this nanomaterial was recognized recently regarding its antimicrobial activity. The particle form, size, and surface chemistry have been recognized as crucial parameters determining the interaction of nanomaterials with biological entities. Furthermore, considering a narrow boundary between beneficial and toxic effects for selenium per se, it is clear that investigations of biomedical applications of SeNPs are very demanding and must be done with great precautions. The goal of this work is to evaluate the effects of SeNPs surface chemistry and structure on antimicrobial activity against several common bacterial strains, including Staphylococcus aureus (ATCC 6538), Enterococcus faecalis (ATCC 29212), Bacillus subtilis (ATCC 6633), and Kocuria rhizophila (ATCC 9341), as well as Escherichia coli (ATCC 8739), Salmonella Abony (NCTC 6017), Klebsiella pneumoniae (NCIMB 9111) and Pseudomonas aeruginosa (ATCC 9027), and the standard yeast strain Candida albicans (ATCC 10231). Three types of SeNPs were synthesized by chemical reduction approach using different stabilizers and reducing agents: (i) bovine serum albumin (BSA) + ascorbic acid, (ii) chitosan + ascorbic acid, and (iii) with glucose. A thorough physicochemical characterization of the obtained SeNPs was performed to determine the effects of varying synthesis parameters on their morphology, size, structure, and surface chemistry. All SeNPs were amorphous, with spherical morphology and size in the range 70–300 nm. However, the SeNPs obtained under different synthesis conditions, i.e. by using different stabilizers as well as reducing agents, exhibited different antimicrobial activity as well as cytotoxicity which are crucial for their applications. In this paper, the antimicrobial screening of the selected systems is presented, which was determined by the broth microdilution method, and inhibitory influence on the production of monomicrobial and dual-species biofilm was evaluated. The potential mechanism of action of different systems is proposed. Additionally, the cytotoxicity of SeNPs was examined on the MRC-5 cell line, in the same concentration interval as for antimicrobial testing. It was shown that formulation SeNPs-BSA expressed a significantly lower cytotoxic effect than the other two formulations.",
publisher = "Frontiers",
journal = "Frontiers in Bioengineering and Biotechnology, Frontiers in Bioengineering and Biotechnology",
title = "Comparative Study of the Antimicrobial Activity of Selenium Nanoparticles With Different Surface Chemistry and Structure",
volume = "8",
doi = "10.3389/fbioe.2020.624621"
}

Filipović, N., Ušjak, D., Milenković, M. T., Zheng, K., Liverani, L., Boccaccini, A. R.,& Stevanović, M. M.. (2021). Comparative Study of the Antimicrobial Activity of Selenium Nanoparticles With Different Surface Chemistry and Structure. in Frontiers in Bioengineering and Biotechnology
Frontiers., 8.
https://doi.org/10.3389/fbioe.2020.624621

Filipović N, Ušjak D, Milenković MT, Zheng K, Liverani L, Boccaccini AR, Stevanović MM. Comparative Study of the Antimicrobial Activity of Selenium Nanoparticles With Different Surface Chemistry and Structure. in Frontiers in Bioengineering and Biotechnology. 2021;8.
doi:10.3389/fbioe.2020.624621 .

Filipović, Nenad, Ušjak, Dušan, Milenković, Marina T., Zheng, Kai, Liverani, Liliana, Boccaccini, Aldo R., Stevanović, Magdalena M., "Comparative Study of the Antimicrobial Activity of Selenium Nanoparticles With Different Surface Chemistry and Structure" in Frontiers in Bioengineering and Biotechnology, 8 (2021),
https://doi.org/10.3389/fbioe.2020.624621 . .

TY  - JOUR
AU  - Ilić, M.
AU  - Samardžić, S.
AU  - Kotur-Stevuljević, J.
AU  - Ušjak, D.
AU  - Milenković, M.
AU  - Kovačević, N.
AU  - Drobac, M.
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1771
AB  - OBJECTIVE: Plants and plant extracts are of great scientific interest due to the
chemical diversity and pharmacological properties of present bioactive molecules. The Geranium L. species are widely used in ethnomedicine. In the current study, the total phenolic and
tannin content, antioxidant and antimicrobial
activity of methanol extracts of eight Geranium
species were investigated.
MATERIALS AND METHODS: The total phenolic and tannin content were determined by the
FC method. Antioxidant capacity was evaluated
in FRAP, DPPH, and biochemical assays, while
antimicrobial activity was examined using the
broth microdilution method.
RESULTS: The high total phenolic (170.64-
636.32 mg GAE/g dry extract) and tannin content (37.80-414.02 mg GAE/g DE), along with significant total antioxidant (FRAP values 1.13-8.80
mmol Fe2+/g) and DPPH radical scavenging activity (SC50 values 4.24-34.52 µg/mL) were observed. The prominent antioxidant capacity was
confirmed in biochemical assays (OS values -1.47
– -13.02). The extracts exhibited significant antimicrobial activity against ATTC strains (MICs
dominantly in the range of 12.5-200 µg/mL) as
well as against clinical isolates of E. coli (MICs
mostly 50 and 100 µg/mL). The pronounced antioxidant and antimicrobial activity can be due to
the high phenolic content, particularly due to the
presence of hydrolyzable tannins.
CONCLUSIONS: Based on the high content of
polyphenols, pronounced antioxidant and antimicrobial activities, the examined extracts are
promising natural antioxidant and antimicrobial
agents with the potential medicinal purpose and
use as a functional food.
PB  - Roma : Verduci Editore
T2  - European Review for Medical and Pharmacological Sciences
T1  - Polyphenol rich extracts of Geranium L. species as potential natural antioxidant and antimicrobial agents
EP  - 6294
IS  - 20
SP  - 6283
VL  - 25
DO  - 10.26355/eurrev_202110_26998
ER  - 

@article{
author = "Ilić, M. and Samardžić, S. and Kotur-Stevuljević, J. and Ušjak, D. and Milenković, M. and Kovačević, N. and Drobac, M.",
year = "2021",
abstract = "OBJECTIVE: Plants and plant extracts are of great scientific interest due to the
chemical diversity and pharmacological properties of present bioactive molecules. The Geranium L. species are widely used in ethnomedicine. In the current study, the total phenolic and
tannin content, antioxidant and antimicrobial
activity of methanol extracts of eight Geranium
species were investigated.
MATERIALS AND METHODS: The total phenolic and tannin content were determined by the
FC method. Antioxidant capacity was evaluated
in FRAP, DPPH, and biochemical assays, while
antimicrobial activity was examined using the
broth microdilution method.
RESULTS: The high total phenolic (170.64-
636.32 mg GAE/g dry extract) and tannin content (37.80-414.02 mg GAE/g DE), along with significant total antioxidant (FRAP values 1.13-8.80
mmol Fe2+/g) and DPPH radical scavenging activity (SC50 values 4.24-34.52 µg/mL) were observed. The prominent antioxidant capacity was
confirmed in biochemical assays (OS values -1.47
– -13.02). The extracts exhibited significant antimicrobial activity against ATTC strains (MICs
dominantly in the range of 12.5-200 µg/mL) as
well as against clinical isolates of E. coli (MICs
mostly 50 and 100 µg/mL). The pronounced antioxidant and antimicrobial activity can be due to
the high phenolic content, particularly due to the
presence of hydrolyzable tannins.
CONCLUSIONS: Based on the high content of
polyphenols, pronounced antioxidant and antimicrobial activities, the examined extracts are
promising natural antioxidant and antimicrobial
agents with the potential medicinal purpose and
use as a functional food.",
publisher = "Roma : Verduci Editore",
journal = "European Review for Medical and Pharmacological Sciences",
title = "Polyphenol rich extracts of Geranium L. species as potential natural antioxidant and antimicrobial agents",
pages = "6294-6283",
number = "20",
volume = "25",
doi = "10.26355/eurrev_202110_26998"
}

Ilić, M., Samardžić, S., Kotur-Stevuljević, J., Ušjak, D., Milenković, M., Kovačević, N.,& Drobac, M.. (2021). Polyphenol rich extracts of Geranium L. species as potential natural antioxidant and antimicrobial agents. in European Review for Medical and Pharmacological Sciences
Roma : Verduci Editore., 25(20), 6283-6294.
https://doi.org/10.26355/eurrev_202110_26998

Ilić M, Samardžić S, Kotur-Stevuljević J, Ušjak D, Milenković M, Kovačević N, Drobac M. Polyphenol rich extracts of Geranium L. species as potential natural antioxidant and antimicrobial agents. in European Review for Medical and Pharmacological Sciences. 2021;25(20):6283-6294.
doi:10.26355/eurrev_202110_26998 .

Ilić, M., Samardžić, S., Kotur-Stevuljević, J., Ušjak, D., Milenković, M., Kovačević, N., Drobac, M., "Polyphenol rich extracts of Geranium L. species as potential natural antioxidant and antimicrobial agents" in European Review for Medical and Pharmacological Sciences, 25, no. 20 (2021):6283-6294,
https://doi.org/10.26355/eurrev_202110_26998 . .

TY  - JOUR
AU  - Ušjak, Dušan
AU  - Dinić, Miroslav
AU  - Novović, Katarina
AU  - Ivković, Branka
AU  - Filipović, Nenad
AU  - Stevanović, Magdalena
AU  - Milenković, Marina T.
PY  - 2021
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1509
AB  - An increasing lack of available therapeutic options against Acinetobacter baumannii urged researchers to seek alternative ways to fight this extremely resistant nosocomial pathogen. Targeting its virulence appears to be a promising strategy, as it offers considerably reduced selection of resistant mutants. In this study, we tested antibiofilm potential of four synthetic chalcone derivatives against A. baumannii. Compound that showed the greatest activity was selected for further evaluation of its antivirulence properties. Real-time PCR was used to evaluate mRNA expression of biofilm-associated virulence factor genes (ompA, bap, abaI) in treated A. baumannii strains. Also, we examined virulence properties related to the expression of these genes, such as fibronectin- and collagen-mediated adhesion, surface motility, and quorum-sensing activity. The results revealed that the expression of all tested genes is downregulated together with the reduction of adhesion and motility. The conclusion is that 2 '-hydroxy-2-methoxychalcone exhibits antivirulence activity against A. baumannii by inhibiting the expression of ompA and bap genes, which is reflected in reduced biofilm formation, adhesion, and surface motility.
PB  - Wiley-V C H Verlag Gmbh, Weinheim
T2  - Chemistry & Biodiversity
T1  - Methoxy-Substituted Hydroxychalcone Reduces Biofilm Production, Adhesion and Surface Motility of Acinetobacter baumannii by Inhibiting ompA Gene Expression
IS  - 1
VL  - 18
DO  - 10.1002/cbdv.202000786
ER  - 

@article{
author = "Ušjak, Dušan and Dinić, Miroslav and Novović, Katarina and Ivković, Branka and Filipović, Nenad and Stevanović, Magdalena and Milenković, Marina T.",
year = "2021",
abstract = "An increasing lack of available therapeutic options against Acinetobacter baumannii urged researchers to seek alternative ways to fight this extremely resistant nosocomial pathogen. Targeting its virulence appears to be a promising strategy, as it offers considerably reduced selection of resistant mutants. In this study, we tested antibiofilm potential of four synthetic chalcone derivatives against A. baumannii. Compound that showed the greatest activity was selected for further evaluation of its antivirulence properties. Real-time PCR was used to evaluate mRNA expression of biofilm-associated virulence factor genes (ompA, bap, abaI) in treated A. baumannii strains. Also, we examined virulence properties related to the expression of these genes, such as fibronectin- and collagen-mediated adhesion, surface motility, and quorum-sensing activity. The results revealed that the expression of all tested genes is downregulated together with the reduction of adhesion and motility. The conclusion is that 2 '-hydroxy-2-methoxychalcone exhibits antivirulence activity against A. baumannii by inhibiting the expression of ompA and bap genes, which is reflected in reduced biofilm formation, adhesion, and surface motility.",
publisher = "Wiley-V C H Verlag Gmbh, Weinheim",
journal = "Chemistry & Biodiversity",
title = "Methoxy-Substituted Hydroxychalcone Reduces Biofilm Production, Adhesion and Surface Motility of Acinetobacter baumannii by Inhibiting ompA Gene Expression",
number = "1",
volume = "18",
doi = "10.1002/cbdv.202000786"
}

Ušjak, D., Dinić, M., Novović, K., Ivković, B., Filipović, N., Stevanović, M.,& Milenković, M. T.. (2021). Methoxy-Substituted Hydroxychalcone Reduces Biofilm Production, Adhesion and Surface Motility of Acinetobacter baumannii by Inhibiting ompA Gene Expression. in Chemistry & Biodiversity
Wiley-V C H Verlag Gmbh, Weinheim., 18(1).
https://doi.org/10.1002/cbdv.202000786

Ušjak D, Dinić M, Novović K, Ivković B, Filipović N, Stevanović M, Milenković MT. Methoxy-Substituted Hydroxychalcone Reduces Biofilm Production, Adhesion and Surface Motility of Acinetobacter baumannii by Inhibiting ompA Gene Expression. in Chemistry & Biodiversity. 2021;18(1).
doi:10.1002/cbdv.202000786 .

Ušjak, Dušan, Dinić, Miroslav, Novović, Katarina, Ivković, Branka, Filipović, Nenad, Stevanović, Magdalena, Milenković, Marina T., "Methoxy-Substituted Hydroxychalcone Reduces Biofilm Production, Adhesion and Surface Motility of Acinetobacter baumannii by Inhibiting ompA Gene Expression" in Chemistry & Biodiversity, 18, no. 1 (2021),
https://doi.org/10.1002/cbdv.202000786 . .

TY  - JOUR
AU  - Ušjak, Dušan
AU  - Ivković, Branka
AU  - Božić, Dragana D.
AU  - Bošković, Lidija
AU  - Milenković, Marina
PY  - 2019
UR  - https://www.sciencedirect.com/science/article/pii/S0882401018319132
UR  - https://imagine.imgge.bg.ac.rs/handle/123456789/1770
AB  - Acinetobacter baumannii and Pseudomonas aeruginosa are frequent multiresistant nosocomial pathogens that cause wound and pulmonary infections in hospitalized patients. As being increasingly resistant to most clinically available antibiotics, there is a constant need for exploration of new substances that could kill them or inhibit their growth, or alternatively inhibit some of their essential virulence factors. Chalcones are chemical compounds with well-documented antimicrobial potential. The aim of this study was to examine effectiveness of four newly-synthesized chalcones against the multiresistant clinical strains of A. baumannii and P. aeruginosa. Antibacterial activity of chalcones was investigated with broth-microdilution test and time-dependent killing assay. Synergistic effects of tested compounds with antibiotics (meropenem, amikacin and ciprofloxacin) were determined by checkerboard assay. The effects of chalcones on expression of virulence factors in P. aeruginosa (pyocyanin production, swimming and swarming motility) and A. baumannii (twitching and surface-associated motility), along with their biofilm production, were also examined. The obtained results indicate substantial antimicrobial activity of the tested chalcones (MICs = 100–175 μg/mL) and several synergistic interactions with antibiotics, as well as notable reduction in expression of all investigated virulence factors. These promising results may constitute a good basis for further research.
T2  - Microbial Pathogenesis
T2  - Microbial PathogenesisMicrobial Pathogenesis
T1  - Antimicrobial activity of novel chalcones and modulation of virulence factors in hospital strains of Acinetobacter baumannii and Pseudomonas aeruginosa
EP  - 196
SP  - 186
VL  - 131
DO  - 10.1016/j.micpath.2019.04.015
ER  - 

@article{
author = "Ušjak, Dušan and Ivković, Branka and Božić, Dragana D. and Bošković, Lidija and Milenković, Marina",
year = "2019",
abstract = "Acinetobacter baumannii and Pseudomonas aeruginosa are frequent multiresistant nosocomial pathogens that cause wound and pulmonary infections in hospitalized patients. As being increasingly resistant to most clinically available antibiotics, there is a constant need for exploration of new substances that could kill them or inhibit their growth, or alternatively inhibit some of their essential virulence factors. Chalcones are chemical compounds with well-documented antimicrobial potential. The aim of this study was to examine effectiveness of four newly-synthesized chalcones against the multiresistant clinical strains of A. baumannii and P. aeruginosa. Antibacterial activity of chalcones was investigated with broth-microdilution test and time-dependent killing assay. Synergistic effects of tested compounds with antibiotics (meropenem, amikacin and ciprofloxacin) were determined by checkerboard assay. The effects of chalcones on expression of virulence factors in P. aeruginosa (pyocyanin production, swimming and swarming motility) and A. baumannii (twitching and surface-associated motility), along with their biofilm production, were also examined. The obtained results indicate substantial antimicrobial activity of the tested chalcones (MICs = 100–175 μg/mL) and several synergistic interactions with antibiotics, as well as notable reduction in expression of all investigated virulence factors. These promising results may constitute a good basis for further research.",
journal = "Microbial Pathogenesis, Microbial PathogenesisMicrobial Pathogenesis",
title = "Antimicrobial activity of novel chalcones and modulation of virulence factors in hospital strains of Acinetobacter baumannii and Pseudomonas aeruginosa",
pages = "196-186",
volume = "131",
doi = "10.1016/j.micpath.2019.04.015"
}

Ušjak, D., Ivković, B., Božić, D. D., Bošković, L.,& Milenković, M.. (2019). Antimicrobial activity of novel chalcones and modulation of virulence factors in hospital strains of Acinetobacter baumannii and Pseudomonas aeruginosa. in Microbial Pathogenesis, 131, 186-196.
https://doi.org/10.1016/j.micpath.2019.04.015

Ušjak D, Ivković B, Božić DD, Bošković L, Milenković M. Antimicrobial activity of novel chalcones and modulation of virulence factors in hospital strains of Acinetobacter baumannii and Pseudomonas aeruginosa. in Microbial Pathogenesis. 2019;131:186-196.
doi:10.1016/j.micpath.2019.04.015 .

Ušjak, Dušan, Ivković, Branka, Božić, Dragana D., Bošković, Lidija, Milenković, Marina, "Antimicrobial activity of novel chalcones and modulation of virulence factors in hospital strains of Acinetobacter baumannii and Pseudomonas aeruginosa" in Microbial Pathogenesis, 131 (2019):186-196,
https://doi.org/10.1016/j.micpath.2019.04.015 . .