Novel cinnamic acid-based PET derivatives as quorum sensing modulators

Poly(ethylene terephthalate) (PET) is widely used material in the healthcare due to its mechanical properties including resistance to chemicals and abrasion. However, it is susceptible to bacterial attachment and contamination. This study addresses some newly designed model compounds of PET with antimicrobial properties that could potentially be incorporated into PET materials. All compounds were synthesized for the first time by labeling an integral part of PET with chromophores in the form of esters of cinnamic and ferulic acids. After complete structural characterization, the effect of new compounds on microbial growth and communication (quorum sensing, QS) was analyzed and further investigated using molecular docking. The obtained results indicate that the introduction of chromophores that have one part of cinnamic acid enriched with a methoxy functional group in them acts as QS modulators. Moreover, compounds exhibited dose-dependent selectivity toward QS signaling pathways and the highest tested concentration of compounds showed Pseudomonas Quinolone Signal (PQS) inhibitory activity suggesting that these compounds have a potential effect on pyocyanin production. Docking studies demonstrated that compounds hold binding power to all four QS protein targets (LuxP, periplasmatic protein that binds AI-2 inducer and forms a complex able to transduce the autoinducer signal, RhIR protein that is a key QS transcriptional regulator that activates the genes involved in the synthesis of rhamnolipids and pyocyanin, AbaI protein that has a role in QS signal transduction, and LasR protein which is a key QS transcriptional regulator that activates transcription of genes coding for some virulence-associated traits) while the highest binding strength is observed with compounds 2 and 6 containing single cinnamic acid fragment, suggesting their further biomedical application

New concept for treatment of glycogen storage disease Ib and diabetes mellitus type 2: small molecule compounds able to adjust glucose level through binding glucose-6-phospate translocase (GlucoAdjust)

GlucoAdjust project aims to solve fast-growing health challenges important for the society, namely it will focus on discovery of new treatments for one rare disease glycogen storage disease type Ib (GSD Ib) and one common disease diabetes mellitus type 2 (DM type 2). We propose a completely new concept based on the identification of small molecule (SM) compounds able to directly bind to glucose-6-phospate translocase (G6PT) and thus fine-tune glucose level. To tackle these challenges, an interdisciplinary international team will screen large library of SMs combining in silico and in vitro approaches and identify SMs that directly bind to G6PT. SMs able to stabilize G6PT and increase its function thus correcting hypoglycemia are potential drugs for GSD Ib. On the other hand, SMs that inhibit G6PT may be used to induce hypoglycemia in DM type 2 treatment. To obtain highly functional results of testing SMs in vitro, human hepatocyte models for GSD Ib and DM type 2 as well as controls will be developed (differentiated from human healthy and GSD Ib iPSC and diabetic adipose stem cells). For the first time, whole transcriptome of human GSD Ib hepatocytes will be used to delineate molecular processes disturbed due to G6PT deficiency. As a result, RNA hallmarks of GSD Ib phenotype will be determined and used for evaluation of SMs’ effect in both models. To be efficient for GSD Ib, SMs will have to revert GSD Ib phenotype into normal. The key to discover satisfactorily effective, yet sufficiently mild inhibitor for DM type 2 will be to avoid hallmarks representing GSD Ib phenotype. Thus, a revolutionary concept of using GSD Ib as a model of hypoglycemia to better optimize DM type 2 treatment is proposed here. Results will be openly disseminated to make a wide scientific, educational, social and economic impacts. GlucoAdjust anticipates innovative results with a potential to be further translated into drugs able to improve lives of people with GSD Ib and DM type 2 worldwide.Principal Investigator:Dr Maja Stojiljkovic, IMGGEDuration period: 2024-202

Silver(I) complexes containing antifungal azoles: significant improvement of the anti-Candida potential of the azole drug after its coordination to the silver(I) ion

Inspired by the emergence of resistance to currently available antifungal therapy and by the great potential of metal complexes for the treatment of various diseases, we synthesized three new silver(I) complexes containing clinically used antifungal azoles as ligands, [Ag(ecz)2]SbF6 (1, ecz is econazole), {[Ag(vcz)2]SbF6}n (2, vcz is voriconazole), and [Ag(ctz)2]SbF6 (3, ctz is clotrimazole), and investigated their antimicrobial properties. The synthesized complexes were characterized by mass spectrometry, IR, UV-vis and 1H NMR spectroscopy, cyclic voltammetry, and single-crystal X-ray diffraction analysis. In the mononuclear complexes 1 and 3 with ecz and ctz, respectively, the silver(I) ion has the expected linear geometry, in which the azoles are monodentately coordinated to this metal center through the N3 imidazole nitrogen atom. In contrast, the vcz-containing complex 2 has a polymeric structure in the solid state in which the silver(I) ions are coordinated by four nitrogen atoms in a distorted tetrahedral geometry. DFT calculations were done to predict the most favorable structures of the studied complexes in DMSO solution. All the studied silver(I) complexes have shown excellent antifungal and good to moderate antibacterial activities with minimal inhibitory concentration (MIC) values in the ranges of 0.01–27.1 and 2.61–47.9 μM on the selected panel of fungi and bacteria, respectively. Importantly, the complexes 1–3 have exhibited a significantly improved antifungal activity compared to the free azoles, with the most pronounced effect observed in the case of complex 2 compared to the parent vcz against Candida glabrata with an increase of activity by five orders of magnitude. Moreover, the silver(I)-azole complexes 2 and 3 significantly inhibited the formation of C. albicans hyphae and biofilms at the subinhibitory concentration of 50% MIC. To investigate the impact of the complex 3 more thoroughly on Candida pathogenesis, its effect on the adherence of C. albicans to A549 cells (human adenocarcinoma alveolar basal epithelial cells), as an initial step of the invasion of host cells, was studied

Assessment of seasonal airborne resistome dynamics in response to air pollution exposure in the Belgrade metropolitan area (AirPollRes)

Antimicrobial resistance (AMR) and air pollution have been identified as one of the most serious threats to human health worldwide. The scarce data demonstrating their interdependence indicates a need to obtain evidence from a broader global area, especially from regions exposed to high air pollution. Considering that Serbia is a country struggling with excessive antibiotics use and misuse, a high percentage of multidrug-resistant bacterial isolates, and poor air quality, the Serbian capital Belgrade has been recognized as an interesting research model for the effects of air pollution on the airborne transmission of AMR. After optimization of air sampling and DNA extraction protocol, the air samples will be collected at nine locations in the Belgrade metropolitan area selected according to air pollution level during four seasons. The state-of-the-art shotgun metagenomic sequencing and bioinformatic analysis of obtained sequences will provide information about microbial community composition of airborne metagenomes. In addition, sequenced airborne metagenomes will be analyzed for the abundance and diversity of resistomes (antibiotic and biocide/metal resistance genes) and mobilomes using several databases and tools. Correlation analyses will offer us insight into the effect of air pollution and seasonal variations on abundance and diversity of airborne pathogens, resistome and mobilome in the Belgrade metropolitan area. In-depth approach of the AirPollRes project will provide the first insights into intersection of AMR and air pollution in the Belgrade metropolitan area, which is highly vulnerable to these health threats. As the AirPollRes is a pioneering project in this field, the expected short-term impact is the introduction of routine monitoring of pathogenic microbes and resistance determinants in the air in Belgrade, while the longer-term impact will be reflected in the improvement of human and animal health, allowing for a longer life with higher quality.Principal Investigator: Dr Katarina Novovic, IMGGEDuration period: 2024-202

Uncovering resistance to microtubule targeting drugs

Drugs that alter microtubule dynamics have been used for decades for treating different types of cancers. Such drugs arrest cell cycle progression in mitosis, and induce apoptosis. However, a fraction of cells manages to survive, escapes from the arrest and resumes proliferation. Understanding the strategies of these cells is important to uncover the early stages of emergence of resistance. To this aim, we performed laboratory evolution experiments in yeast and in mammalian cells when microtubule dynamics is impaired. Our results show that cells follow reproducible strategies to escape the effect of the drug. Via mutations, aneuplouidy and non genetics mechanisms they recover microtubule functionality and decrease the propensity to die.Book of abstract: 4th Belgrade Bioinformatics Conference, June 19-23, 202

Interakcije eNOS i ACE gena i duvanskog dima u hroničnoj opstruktivnoj bolesti pluća

Background: Chronic obstructive pulmonary disease(COPD) is a complex disorder with unexplained heritability.Interactions of genetic and environmental factors arethought to be crucial in COPD. So, we aim to examineinteractions of the endothelial nitric oxide synthase (eNOS)and angiotensin converting enzyme (ACE) genes and ciga-rette smoking in COPD. Methods:The eNOS G894T and ACE ID variants wereanalyzed in 122 COPD patients and 200 controls fromSerbia. The effect of the variants on COPD was assessed bylogistic regression. Interactions between eNOS, ACE andcigarette smoking in COPD were evaluated using a case-control model. Interaction between the genes was analyzedin silico. Results:No effect of the eNOS G894T and ACE ID variantson COPD was found in our study. Gene-gene interactionbetween the eNOS TT and ACE D was identified(p=0.033) in COPD. The interaction is realized within the complex network of biochemical pathways. Gene-environ-ment interactions between the eNOS T and cigarettesmoking (p=0.013), and the ACE II and cigarette smoking(p=0.009) were detected in COPD in our study. Conclusions:This is the first research to reveal interactionsof the eNOS and ACE genes and cigarette smoking inCOPD progressing our understanding of COPD heritabilityand contributing to the development of appropriate treat-ments.Uvod: Hronična opstruktivna bolest pluća (HOBP) je složeno oboljenje sa nerazjašnjenom genetičkom osnovom.Smatra se da su interakcije genetskih i spoljašnjih faktora ključne u HOBP. Stoga je naš cilj bio da ispitamo interakcijegena za endotelijalnu azot-monoksid sintazu (eNOS) i angiotenzin konvertujući enzim (ACE) i duvanskog dima uHOBP. Metode :eNOS G894T i ACE ID varijante su analiziranekod 122 HOBP pacijenta i 200 kontrola iz Srbije. Uticajvarijanti na HOBP je ispitan logističkom regresijom. Interakcije izme|u eNOS, ACE i duvanskog dima u HOBP suispitane korišćenjem modela slučaj-kontrola. Interakciaja između gena je analizirana in silico. Rezultati: Prema ovoj studiji eNOS G894T i ACE ID varijante nemaju uticaj na HOBP. Gen-gen interakcija između eNOS TT i ACE D je identifikovana (p=0,033) u HOBP.Ova interakcija se ostvaruje u okviru složene mreže bio-hemijskih puteva. Gen-sredina interakcije izme|u eNOS Ti duvanskog dima (p=0,013), i ACE II i duvanskog dima(p=0,009) su detektovane u HOBP u ovoj studiji.Zaključak: Ovo je prvo istraživanje u kome su otkriveneinterakcije između eNOS i ACE gena i duvanskog dima uHOBP što doprinosi našem razumevanju genetičke osnove HOBP i razvoju adekvatnog tretmana

Can we use biobanks to study infectious diseases?

Understanding the molecular and environmental basis of diseases in order to improve diagnosis and treatment represent a top priority for researchers. Much of the progress occurred following the growth of various omics technologies and the IT progress in developing large electronic databases capable of storing huge amounts of data. Biobanks represents the most valuable resource for personalized medicine as these are the large collection of various patient samples with well-annotated clinical data which strive to identify possible links between genetic predisposition and disease. A significant step forward are biobanks that are linked to the electronic health records of each participant enabling up-to-date source of relevant medical information and those “deeply phenotyped” for various other omics data, such as microbiome, epigenome, transcriptome, metabolome and proteome. Since infectious diseases still represent a huge threat to global human health, and host genetic factors have been implied as determining risk factors for observed variations in disease susceptibility, severity, and outcome, during this lecture we will discuss challenges and opportunities of using biobanks as a potential source to study infectious diseases based on the case example of isolated population-based longitudinal biobank “10,001 Dalmatians”. Results of a genome-wide association meta-analyses of 14 different infectious-related phenotypes identified 29 infection-related genetic associations, most belonging to rare variants, all of which have a role in immune response. These findings support the concept that host genetic susceptibility to bacterial and viral infections in adults is polygenic, where common variations have very low explained variance and/or “unfortunate” combinations of numerous rare variants. Expanding our understanding of rare variants may help in the construction of genetic panels which might predict an individual’s lifetime vulnerability to major infectious diseases. Furthermore, longitudinal biobanks are a valuable source of data for discovering host genetic variations involved in infectious disease susceptibility and severity. Because infectious diseases continue to exert selective pressure on our genomes, a global network of biobanks with access to genetic and environmental data is required to further explain complicated mechanisms underlying host-pathogen interactions and infectious disease vulnerability.Book of abstract: 4th Belgrade Bioinformatics Conference, June 19-23, 202

Supplementary material for: Spasovski, V.; Srzentić Dražilov, S.; Nikčević, G.; Baščarević, Z.; Stojiljković, M.; Pavlović, S.; Spasovski, D. Molecular Biomarkers in Perthes Disease: A Review. Diagnostics 2023, 13 (3), 471. https://doi.org/10.3390/diagnostics13030471.

Background: Perthes disease is a juvenile form of osteonecrosis of the femoral head that affects children under the age of 15. One hundred years after its discovery, some light has been shed on its etiology and the biological factors relevant to its etiology and disease severity. Methods: The aim of this study was to summarize the literature findings on the biological factors relevant to the pathogenesis of Perthes disease, their diagnostic and clinical significance, and their therapeutic potential. A special focus on candidate genes as susceptibility factors and factors relevant to clinical severity was made, where studies reporting clinical or preclinical results were considered as the inclusion criteria. PubMed databases were searched by two independent researchers. Sixty-eight articles were included in this review. Results on the factors relevant to vascular involvement and inflammatory molecules indicated as factors that contribute to impaired bone remodeling have been summarized. Moreover, several candidate genes relevant to an active phase of the disease have been suggested as possible biological therapeutic targets. Conclusions: Delineation of molecular biomarkers that underlie the pathophysiological process of Perthes disease can allow for the provision of earlier and more accurate diagnoses of the disease and more precise follow-ups and treatment in the early phases of the disease.Supplementary material for: [https://doi.org/10.3390/diagnostics13030471]Related to the published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1853

Stability and bioactive compounds assessment of yogurt containing novel natural starter cultures with the ability to promote longevity in Caenorhabditis elegans

Yogurt represent one of the oldest fermented foods containing viable lactic acid bacteria and many bioactive compounds that could exhibit beneficial effects on human health and train our immune system to better respond to invading pathogens. Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus are commonly used for yogurt preparation under controlled temperature and environmental conditions. In this study, we investigated probiotic features of S. thermophilus BGKMJ1-36 and L. bulgaricus BGVLJ1-21 strains isolated from artisanal sour milk and yogurt by using Caenorhabditis elegans as an in vivo model system. Further, we evaluated content of total fat, saturated fatty acids, proteins, and lactose, as well as vitamins and AA of yogurt prepared from above-mentioned starter cultures during 21 d of storage at 4°C to get insights of final product stability. We showed that S. thermophilus BGKMJ1-36 and L. bulgaricus BGVLJ1-21 strains applied in combination upregulated the expression of autophagy-related genes in C. elegans. Beside autophagy, we observed activation of TIR-1-dependent transcription of lysozyme-like antimicrobial genes involved in the immune defense of C. elegans. Upregulation of these genes strongly correlates with an increase in the longevity of the worms fed with yogurt culture bacteria. Further, we showed that yogurt prepared with S. thermophilus BGKMJ1-36 and L. bulgaricus BGVLJ1-21, as a final product, is rich with vitamin B2 and dominant AA known by their prolongevity properties. Taken together, our study pointed to the beneficial features of the tested starter cultures and yogurt and highlighted their potential to be used as a fermented food with added-value properties

Determination of muscle fiber types expressing ANKRD2

Introduction: Ankyrin Repeat Domain 2 (ANKRD2) is expressed in skeletal muscle, where plays a role in muscle development, differentiation and adaptation to stress. Human skeletal muscle consists of three major fiber types: type 1 (slow-twitch, oxidative), type 2A (fast-twitch, oxidative) and type 2X (fast-twitch, glycolytic). ANKRD2 is reported to be primarily expressed in type 1 myofibers. However, recent findings on human single myofibers and our study of chicken muscles have shown that this protein may also be expressed in type 2A fibers. Hence, our objective was to examine whether ANKRD2 is present in human fast, type 2A muscle fibers using immunohistochemistry. Methods: Samples of large leg musclessoleus, gastrocnemius, vastusintermedius and vastuslateralis were obtained from human cadaveric tissue. Serial cryosections were independently stained with anti-ANKRD2 and antibodies for different myosin heavy chain isoforms (6H1 for type 2X, BF35 for type 1 and 2A, antiMHCs for type 1 and anti-MHCf for type 2A and 2X fibers). Immunostained tissues were analyzed by fluorescent microscopy. Results: In addition to slow, type 1, ANKRD2 wasfound expressed in fast, type 2A myofibers, which both have oxidative metabolism. Further, we did not observe ANDRD2 expression in glycolytic, type 2X myiofibers. This pattern of ANKRD2 expression was consistent across all examined muscles. Conclusion: Our resultsimplicate that the regulatory mechanism of ANKRD2 expression in human skeletal muscle is associated with oxidative metabolism, rather than muscle contraction speed