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Utilization of supercritical carbon dioxide in fabrication of cellulose acetate films with anti-biofilm effects against Pseudomonas aeruginosa and Staphylococcus aureus
No full textThis study discusses utilization of supercritical carbon dioxide for impregnation of cellulose acetate films with thymol in order to produce material with anti-biofilm activity against Pseudomonas aeruginosa and Staphylococcus aureus. Analysis of anti-biofilm activity of cellulose acetate beads impregnated with thymol suggested that optimal thymol loading was in the range from 26% to 30% for efficient reduction of biofilm formation and eradication of pre-formed biofilms. Polymer films were fabricated by the solvent casting method from polymer solutions of different contents, and loaded with thymol using supercritical carbon dioxide at 15.5 MPa and 35 degrees C. The film containing 30% of thymol (F1 30%) exhibited substantial anti-adhesion properties inhibiting biofilm formation on its surface and considerably reduced formation of biofilms on the surrounding surfaces (up to 80%) by all tested strains including antibiotic resistant P. aeruginosa DM50 and methicillin-resistant S. aureus.Peer-reviewed manuscript: [https://imagine.imgge.bg.ac.rs/handle/123456789/1627
Microbial Production of Violacein and Process Optimization for Dyeing Polyamide Fabrics With Acquired Antimicrobial Properties
Get PDFIn the present study, crude bacterial extract containing violacein is investigated for the preparation of antimicrobial polyamide fabrics. The optimal culture conditions of Janthinobacterium lividum (JL) for maximum biomass and violacein production were found to be 25 degrees C, pH 7.0, while the addition of ampicillin of 0.2 mg mL(-1) in the small scale increased violacein production 1.3-fold. In scale-up trials, the addition of 1% (v/v) glycerol in a fed-batch bioreactor, resulted in fivefold extracted crude violacein increase with final concentration of 1.828 g L-1. Polyamide 6.6 fabrics were dyed following three different processes; through simultaneous fermentation and dyeing (SFD), by incubating the fabric in the sonicated bacterial culture after fermentation and by using cell-free extract containing violacein. Maximum color change (Delta E) and color strength (K/S) obtained for SFD fabrics were 74.81 and 22.01, respectively, while no alteration of fastness and staining of dye at acid and alkaline perspiration or at water was indicated. The dyed fabrics presented significant antifungal activity against Candida albicans, C. parapsilosis, and C. krusei, as well as antibacterial properties against Escherichia coli, Staphylococcus aureus, and the S. aureus MRSA. We have shown that J. lividum cultures can be successfully used for violacein production and for simultaneous dying of fabrics resulting in dyed fabrics with antimicrobial properties without utilization of organic solvents
Comparative phylogeography of capitulate Campanula species from the Balkans, with description of a new species, C-daucoides
No full textCapitulate inflorescence is a specific, strongly adaptive and rare feature in the genus Campanula. We studied morphologically eight capitulate Campanula taxa from the Balkans (1537 individuals/52 populations) and one more species from Caucasus at the molecular level (using chloroplast markers trnG(UCC)-trnS(GCU) and psbA-trnH, 130 individuals/58 populations) to assess their relations and evolutionary histories. Although all studied taxa were well circumscribed at both the morphological and molecular levels (except morphologically distinct but genetically invariable C. moesiaca which acquired its single haplotype via past cytoplasmic introgression from C. cervicaria), their relations inferred from the two datasets were incongruent possibly due to the homoplasy of morphological characters frequently reported in Campanula. Interspecific hybridization and introgression affected majority of studied species and may be more common in Campanula than previously thought. These processes, along with incomplete lineage sorting and retention of ancestral polymorphisms, hampered our phylogenetic reconstructions and prevented us to fully resolve species relations, and to support monophyletic origin of capitulate Campanula species. Nonetheless, several cryptic taxa were delineated, and C. daucoides was described as a new capitulate Campanula species. Different evolutionary histories and multiple glacial refugia were inferred for all species represented by multiple samples (except C. moesiaca). According to our dating, their speciation was in most cases triggered by various geo-historic events such as the uplift of the Alpide belt, Messinian Salinity Crisis, or desiccation of the Pannonian Sea/Pliocene Lakes from the central Balkans, while their further diversification was mainly driven by the onset of the Quaternary and cycles of glacials/interglacials
Identification of molecular genetic markers of acute myeloid leukemia pathogenesis
Get PDFAkutna mijeloidna leukemija (AML) predstavlja grupu hematoloških malignih oboljenja koju karakteriše heterogeni genetički i epigenetički profil. AML sa normalnim kariotipom (AML-NK) predstavlja najveću podgrupu u okviru AML, koju odlikuje intermedijarni rizik. Međutim, samo 40% obolelih od AML-NK preživljava duže od 5 godina, pa je neophodno otkriti molekularno-genetičke markere značajne za prognozu i praćenje toka bolesti, rano otkrivanje minimalne rezidualne bolesti (MRB) i izbor odgovarajuće terapije, kako bi bila omogućena bolja stratifikacija pacijenata sa AML-NK. Prvi cilj ove studije je analiza mutacionog profila dečje (dAML) i adultne (aAML) koja će doprineti rasvetljenju patogeneze mijeloidnih leukemija. Drugi cilj predstavlja određivanje nivoa ekspresije WT1, EVI1, BAALC i MN1 gena, kao potencijalnih molekularno-genetičkih markera značajnih za AML-NK. Analizirani su neselektovani DNK uzorci 20 dAML i 20 aAML pacijenata, primenom metode targetovanog sekvenciranja nove generacije (eng. „targeted next generation sequencing“ – NGS), TruSeq Amplicon - Cancer Panel (TSACP) esejem, koji omogućava detekciju somatskih mutacija u 48 gena uključenih u kancerogenezu. U drugom delu ove studije nivo ekspresije gena WT1 (104 pacijenta), EVI1 (104 pacijenta), BAALC (111 pacijenata) i MN1 (111 pacijenata) u mononuklearnim ćelijama koštane srži AML-NK pacijenata i zdravih pojedinaca analiziran je real-time PCR metodom. Rezultati su pokazali da AML sadrži relativno mali broj genetičkih promena, u proseku samo 3 mutacije po pacijentu kod aAML i dAML grupe pacijenata. Zastupljenost najčešćih mutacija u pojedinačnim genima, koji su povezani sa patogenezom AML, razlikovala se kod dAML i aAML: IDH1 (0% kod dAML, 5% kod aAML), IDH2 (0% kod dAML, 10% kod aAML), NPM1 (10% kod dAML, 35% kod aAML). Takođe, prisustvo mutacija u 4 gena (JAK3, ABL1, GNAQ i EGFR) koji kodiraju tirozin kinaze ili proteine koji su udruženi sa tirozin kinazama detektovano je isključivo kod dAML pacijenata, dok je kod 5 gena (IDH1, APC, HNF1A, GNAS i SMARCB1) koji su uključeni u metilaciju i modifikaciju histona prisustvo mutacija utvrđeno isključivo kod aAML pacijenata...Acute myeloid leukemia (AML) is a group of haematological malignancies characterized by a heterogeneous genetic and epigenetic profile. AML with normal karyotype (AML-NK) represents the largest subgroup within the AML, characterized by an intermediate risk. However, only 40% of patients with AML-NK survive for more than 5 years, so it is necessary to detect molecular genetic markers significant for prognosis and disease monitoring, early detection of minimum residual disease (MRD), and the selection of appropriate therapy in order to enable better stratification of patients with AML-NK. The first aim of this study is the analysis of the mutation profile of childhood (cAML) and adult (aAML) AML which will contribute to the clarification of the pathogenesis of myeloid leukemia. The second aim is to determine the level of expression of WT1, EVI1, BAALC and MN1 gene, as potential molecular genetic markers relevant to AML-NK. Unselected DNA samples of 20 cAMLs and 20 aAML patients were analyzed using the targeted next generation sequencing (NGS) method, on the TruSeq Amplicon-Cancer Panel (TSACP), which enables the detection of somatic mutations in 48 genes involved in carcinogenesis. In the second part of this research, the level of expression of the genes WT1 (104 patients), EVI1 (104 patients), BAALC (111 patients) and MN1 (111 patients) was analyzed in mononuclear bone marrow cells of AML-NK patients and healthy individuals by the real-time PCR method. The results showed that the AML contains a relatively small number of genetic changes, on average only 3 mutations per patient in the aAML and cAML group. The incidence of the most common mutations in individual genes, which are associated with AML pathogenesis, differed in cAML and aAML: IDH1 (0% in cAML, 5% in aAML), IDH2 (0% in cAML, 10% in aAML), NPM1 (10% in cAML, 35% in aAML). Also, the presence of mutations in 4 genes (JAK3, ABL1, GNAQ, and EGFR) that encode tyrosine kinases or proteins that are associated with tyrosine kinases were detected exclusively in cAML patients, while in 5 genes (IDH1, APC, HNF1A, GNAS and SMARCB1) involved in methylation and histone modification, the presence of mutations was found exclusively in aAML patients..
Analysis of variants in the promoter region of CYP2C19 gene and their role in prediction of clopidogrel response in patients with myocardial infarcition and carotid stenosis
Get PDFKlopidogrel je antiagregacioni lek koji je sa aspirinom, u sklopu dvojne antiagregacione terapije, indikovan u stanjima koja nose rizik od ishemijskih događaja i tromboembolijskih komplikacija. Postoji velika interindividualna varijabilnost u odgovoru na klopidogrel. Značajan broj pacijenata kao posledicu neadekvatnog odgovora na terapiju može iskusiti različita neželjena dejstva. Kod oko 9% pacijenata koji su na terapiji klopidogrelom javlja se krvarenje kao neželjeni efekat. Ono samo po sebi može biti životno ugrožavajuće, a sekundarno može biti povezano sa pojavom ishemijskih događaja. Uzroci varijabilnosti u odgovoru na terapiju klopidogrelom, mogu biti kako stečeni, tako i posledica genetičkih faktora. Među genetičkim faktorima najznačajniji doprinos ima gen za enzim CYP2C19 koji je neophodan u procesu konverzije klopidogrela u aktivnu formu leka. Zbog toga su varijante CYP2C19 gena najčešće razmatrane u okviru farmakogenetike klopidogrela. Od posebnog kliničkog značaja su varijante rs4244285 (c.681G>A; CYP2C19*2) i rs4986893 (c.636G>A; CYP2C19*3), koje su povezane sa oslabljenim odgovorom na klopidogrel. Promotorska varijanta rs12248560 (c.-806C>T, CYP2C19*17) se dovodi u vezu sa pojačanim stepenom inhibicije trombocita i povišenim rizikom od krvarenja kod pacijenata na terapiji klopidogrelom. U fokusu ove studije bilo je ispitivanje uticaja CYP2C19 gena na pojavu pojačanog odgovora na klopidogrel koje vodi povišenom riziku od krvarenja. Istraživanje je sprovedeno u grupi pacijenata koji su pretrpeli infarkt miokarda i bili podvrgnuti perkutanoj koronarnoj intervenciji (PCI), kao i grupi pacijenata sa stenozom karotida koji su podvrgnuti karotidnoj endarterektomiji (CEA). U studiji su analizirane promotorske varijante rs12248560 (c.-806C>T, CYP2C19*17) i rs11568732 (c.-889T>G, CYP2C19*20) za koje su literaturni podaci i naši preliminarni rezultati pokazali da mogu biti povezane sa pojačanim odgovorom na klopidogrel. Dodatno su analizirane egzonske varijante rs4244285 (c.681G>A; CYP2C19*2) i rs4986893 (c.636G>A, CYP2C19*3) zbog mogućeg uticaja na efekte promotorskih varijanti. Zbog oskudnosti podataka koji se odnose na promotorsku varijantu rs11568732 (c.-889T>G, CYP2C19*20), jedan od ciljeva studije bio je da se u in vitro uslovima ispita njen funkcijski značaj...Wide inter-individual variability in clopidogrel response has been reported. Significant number of patients with inadequate response to clopidogrel experience different adverse effects. Bleeding as an adverse event occurs in about 9% of patients on clopidogrel therapy. It can be life threatening and secondarily, related to higher risk of ischemic events. Variability in clopidogrel response may be explained by different non-genetic and genetic factors. Among genetic factors, the most important role has CYP2C19 gene encoding enzyme responsible for conversion of clopidogrel to its active form. Hence, CYP2C19 gene variants are the most commonly investigated genetic variants related to clopidogrel pharmacogenetics. Clinically, the most relevant variants are rs4244285 (c.681G>A; CYP2C19*2) and rs4986893 (c.636G>A; CYP2C19*3) which lead to clopidogrel poor response. Additionally, promoter variant rs12248560 (c.-806C>T, CYP2C19*17) is associated with elevated platelet inhibition and increased risk of bleeding in patients on clopidogrel. This study was focused on investigating role of CYP2C19 gene in clopidogrel hyper-responsiveness which increases the risk of bleeding. This study was conducted in a group of patients with acute myocardial infarction who underwent percutaneous coronary intervention (PCI) as well as in patients with carotid stenosis who underwent carotid endarterectomy (CEA). In this study we examined two promoter rs12248560 (CYP2C19*17) and rs11568732 (CYP2C19*20) variants, for which literature and our preliminary results showed to be associated with elevated clopidogrel response. Additionally, we analyzed exonic variants rs4244285 (c.681G>A; CYP2C19*2) and rs4986893 (c.636G>A, CYP2C19*3) considering their potential influence on the promoter variants’ effects. Since literature data on rs11568732 (c.-889T>G, CYP2C19*20) variant is scarce, we aimed to investigate its functional significance. Further, one of our aims was the anlysis of the relevant CYP2C19 variants regarding clopidogrel therapy, which can be used to rationalize current protocols for antiplatelet therapy in Serbia..
Molecular basis of colorectal carcinogenesis
No full textКолоректална карциногенеза је вишестепени процес током кога долази до
патолошких неопластичних промена цревне мукозе и акумулације генетичких и
епигенетичких промена у ћелијама цревног епитела. Кључни фактори у настанку
колоректалног карцинома су онкогени и тумор-супресорски гени чије промене доводе до
дерегулације ћелијског циклуса, процеса који лежи у основи карциногенезе. Онкогени
најчешће погођени мутацијама у колоректалном карциному су KRAS и BRAF, док је
мутацијама најчешће инактивиран тумор-супресорски ген APC. Резултати новијих
истраживања указују да битну улогу у карциногенези имају молекули микроРНК, који
дејство остварују посредно, кроз регулацију онкогена и тумор-супресорских гена.
Молекуларни механизми који леже у основи малигне трансформације и настанка
колоректалног карцинома још увек нису у потпуности разјашњени. Открића до којих се
дошло у последњих неколико година, а посебно резултати обимних студија које су
реализоване уз помоћ модерних метода геномике и транскриптомике анализом великог
броја узорака туморског ткива, допринела су проширивању класичног модела
колоректалне карциногенезе у складу са новим сазнањима. Откривене су мутације у
бројним онкогенима и тумор-супресорским генима чији поремећаји могу довести до
настанка или прогресије тумора, као и у различитим другим молекулима који могу да
модулирају овај процес и тако утичу на карактеристике тумора, презентацију и тежину
болести. Новије студије спроведене захваљујући увођењу напредних технологија у
молекуларно-биолошка истраживања указала су на постојање четири молекуларна типа
колоректалног карцинома, при чему сваки има јединствене биолошке одлике и
специфичан профил генске експресије. Осим очигледног доприноса у сфери клиничке
примене, откриће молекуларних подтипова колоректалног карцинома представља основу
за даља истраживања механизама колоректалне карциногенезе која ће бити усмерена на
разјашњење улоге појединих молекула и процеса.Колоректална карциногенеза је вишестепени процес током кога долази до
патолошких неопластичних промена цревне мукозе и акумулације генетичких и
епигенетичких промена у ћелијама цревног епитела. Кључни фактори у настанку
колоректалног карцинома су онкогени и тумор-супресорски гени чије промене доводе до
дерегулације ћелијског циклуса, процеса који лежи у основи карциногенезе. Онкогени
најчешће погођени мутацијама у колоректалном карциному су КРАС и БРАФ, док је
мутацијама најчешће инактивиран тумор-супресорски ген АПЦ. Резултати новијих
истраживања указују да битну улогу у карциногенези имају молекули микроРНК, који
дејство остварују посредно, кроз регулацију онкогена и тумор-супресорских гена.
Молекуларни механизми који леже у основи малигне трансформације и настанка
колоректалног карцинома још увек нису у потпуности разјашњени. Открића до којих се
дошло у последњих неколико година, а посебно резултати обимних студија које су
реализоване уз помоћ модерних метода геномике и транскриптомике анализом великог
броја узорака туморског ткива, допринела су проширивању класичног модела
колоректалне карциногенезе у складу са новим сазнањима. Откривене су мутације у
бројним онкогенима и тумор-супресорским генима чији поремећаји могу довести до
настанка или прогресије тумора, као и у различитим другим молекулима који могу да
модулирају овај процес и тако утичу на карактеристике тумора, презентацију и тежину
болести. Новије студије спроведене захваљујући увођењу напредних технологија у
молекуларно-биолошка истраживања указала су на постојање четири молекуларна типа
колоректалног карцинома, при чему сваки има јединствене биолошке одлике и
специфичан профил генске експресије. Осим очигледног доприноса у сфери клиничке
примене, откриће молекуларних подтипова колоректалног карцинома представља основу
за даља истраживања механизама колоректалне карциногенезе која ће бити усмерена на
разјашњење улоге појединих молекула и процеса.Colorectal carcinogenesis is a multistep process characterized by pathological neoplastic
changes of the intestinal mucosa and the accumulation of genetic and epigenetic changes in the
intestinal epithelial cells. Key factors in the formation of colorectal cancer are oncogenes and
tumor suppressor genes and their alterations lead to deregulation of the cell cycle, the process
underlying carcinogenesis. Oncogenes most commonly affected by mutations in colorectal
cancer are KRAS and BRAF, while the most oftenly inactivated tumor suppressor gene is APC.
The results of recent studies indicate that microRNA molecules play an important role in
carcinogenesis, exerting their effect indirectly through regulation of oncogenes and tumor
suppressor genes. Molecular mechanisms underlying malignant transformation and colorectal
cancer have not yet been fully clarified. The findings of the last few years, and in particular the
results of extensive studies carried out using modern methods of genomics and transcriptomics
by analyzing a large number of tumor tissue samples, have contributed to the extension of the
classic model of colorectal carcinogenesis in line with new findings. Mutations have been
discovered in numerous oncogenes and tumor suppressor genes that can lead to tumor formation
or progression, as well as in various other molecules that can modulate this process and thus
affect the tumor characteristics, presentation, and severity of the disease. Recent studies
conducted by the advanced technologies introduced in molecular biological research have
indicated the existence of four molecular subtypes of colorectal cancer, each having unique
biological features and a specific gene expression profile. In addition to the obvious contribution
in the field of clinical application, the discovery of molecular colorectal cancer subtypes will
form the basis for further research into colorectal carcinogenesis mechanisms that will focus on
clarifying the role of particular molecules and processes
Supplementary material for the article: Savić, N. D.; Vojnovic, S.; Glišić, B. Đ.; Crochet, A.; Pavic, A.; Janjić, G. V.; Pekmezović, M.; Opsenica, I. M.; Fromm, K. M.; Nikodinovic-Runic, J.; et al. Mononuclear Silver(I) Complexes with 1,7-Phenanthroline as Potent Inhibitors of Candida Growth. Eur. J. Med. Chem. 2018, 156, 760–773. https://doi.org/10.1016/j.ejmech.2018.07.049
Get PDFSupplementary data for: [https://doi.org/10.1016/j.ejmech.2018.07.049]Research data for this article: [https://www.ccdc.cam.ac.uk/structures/search?id=doi:10.5517/ccdc.csd.cc1zdr7c&sid=DataCite]Related to published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1098]Related to accepted version: [https://imagine.imgge.bg.ac.rs/handle/123456789/2228
Kinetic profile of the antioxidant activity of propolis extract: 2,2-diphenyl-1-picrylhydrazyl radical bleaching assay
Get PDFThe objective of this paper was to assess the antiradical effectiveness of propolis extract (PE) based on 2.2-diphenyl-1-picrylhydrazyl radical (DPPH center dot) bleaching assay kinetic profile. The kinetic profile of scavenging DPPH center dot for PE exhibited one kinetic period characterized by one kinetic constant. The second-order rate constant (k(2)) for the oxidation of PE by DPPH center dot, determined for the first time in this study, was 0.17 dm(3)g(-1)s(-1). The obtained k(2) value was compared to that of synthetic antioxidants and natural extracts used in the food industry. Kinetic analysis of PE antiradical effectiveness showed that the k(2) was within the range values for natural colorants of fruit extracts and should be considered as a fast acting natural antioxidant source. The k(2) parameter indicates the extent of oxidation inhibition that is based on all of the kinetic profiles of DPPH bleaching rather than single point measurements. For this reason, the kinetic analysis should become a necessary step for more precise antioxidative characterization of propolis
Characterization of zebrafish (Danio rerio) muscle ankyrin repeat proteins reveals their conserved response to endurance exercise
Get PDFMuscle proteins with ankyrin repeats (MARPs) ANKRD1 and ANKRD2 are titin-associated proteins with a putative role as transcriptional co-regulators in striated muscle, involved in the cellular response to mechanical, oxidative and metabolic stress. Since many aspects of the biology of MARPs, particularly exact mechanisms of their action, in striated muscle are still elusive, research in this field will benefit from novel animal model system. Here we investigated the MARPs found in zebrafish for protein structure, evolutionary conservation, spatiotemporal expression profiles and response to increased muscle activity. Ankrd1 and Ankrd2 show overall moderate conservation at the protein level, more pronounced in the region of ankyrin repeats, motifs indispensable for their function. The two zebrafish genes, ankrd1a and ankrd1b, counterparts of mammalian ANKRD1/Ankrd1, have different expression profiles during first seven days of development. Mild increase of ankrd1a transcript levels was detected at 72 hpf (1.74 +/- 0.24 fold increase relative to 24 hpf time point), while ankrd1b expression was markedly upregulated from 24 hpf onward and peaked at 72 hpf (92.18 +/- 36.95 fold increase relative to 24 hpf time point). Spatially, they exhibited non-overlapping expression patterns during skeletal muscle development in trunk (ankrd1a) and tail (ankrd1b) somites. Expression of ankrd2 was barely detectable. Zebrafish MARPs, expressed at a relatively low level in adult striated muscle, were found to be responsive to endurance exercise training consisting of two bouts of 3 hours of forced swimming daily, for five consecutive days. Three hours after the last exercise bout, ankrd1a expression increased in cardiac muscle (6.19 +/- 5.05 fold change), while ankrd1b and ankrd2 were upregulated in skeletal muscle (1.97 +/- 1.05 and 1.84 +/- 0.58 fold change, respectively). This study provides the foundation to establish zebrafish as a novel in vivo model for further investigation of MARPs function in striated muscle
Epidermal Growth Factor Receptor Gene in Non-Small-Cell Lung Cancer: The Importance of Promoter Polymorphism Investigation
Get PDFRecently, epidermal growth factor receptor (EGFR) was a key molecule in investigation of lung cancer, and it was a target for a new therapeutic strategy, based on molecular analyses. In this review, we have summarized some issues considering the role of EGFR in lung cancer, its coding gene, and its promoter gene polymorphisms (SNPs) -216G/T and -191C/A in non-small-cell lung cancer (NSCLC). The position of the SNPs indicates their significant role in EGFR regulation. The accumulation of knowledge regarding SNPs lately suggests their significant and important role in the onset of carcinogenesis, the prediction of the onset of metastases, the response to therapy with TKI inhibitors, and the onset of toxic effects of the applied therapy. Based on this, we suggest further studies of the relationship of clinical significance to SNPs in patients with lung tumors