imagine (Institute of molecular genetics and genetic engineering)
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The first nationwide multicenter study ofAcinetobacter baumanniirecovered in Serbia: emergence of OXA-72, OXA-23 and NDM-1-producing isolates
Get PDFBackground The worldwide emergence and clonal spread of carbapenem-resistantAcinetobacter baumannii(CRAB) is of great concern. The aim of this nationwide study was to investigate the prevalence of CRAB isolates in Serbia and to characterize underlying resistance mechanisms and their genetic relatedness. Methods Non-redundant clinical samples obtained from hospitalized patients throughout Serbia were included in the prospective, observational, multicenter study conducted from January to June 2018. Samples were initially screened for the presence ofAcinetobacter baumannii-calcoaceticus(Acb) complex using conventional bacteriological techniques. Acb complexes recovered from clinical samples obtained from inpatients with confirmed bacterial infections were further evaluated for the presence ofA. baumannii. Identification to the species level was done by the detection of thebla(OXA-51)gene andrpoBgene sequence analysis. Susceptibility testing was done by disk diffusion and broth microdilution method. CRAB isolates were tested for the presence of acquired carbapenemases(bla(OXA-24-like),bla(OXA-23-like,)bla(OXA-58-like),bla(OXA-143-like),bla(IMP),bla(VIM),bla(GIM),bla(SPM),bla(SIM),bla(NDM)) by PCR. Clonal relatedness was assessed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Results Acb complex was isolated in 280 out of 2401 clinical samples (11.6%). Overall,A. baumanniiwas identified in 237 out of 280 Acb complex (84.6%). CRAB prevalence was found to be 93.7% (237/222). The MIC50/MIC(90)for imipenem and meropenem were 8/ gt 32 mu g/mL and 16/ gt 32 mu g/mL, respectively. Although susceptibility was high for colistin (95.7%;n = 227) and tigecycline (75.1%;n = 178), ten isolates (4.3%) were classified as pandrug-resistant. The following carbapenemases-encoding genes were found: 98 (44.2%)bla(OXA-24-like), 76 (34.5%)bla(OXA-23-like), and 7 (3.2%)bla(NDM-1). PFGE analysis revealed six different clusters. MLST analysis identified three STs: ST2 (n = 13), ST492 (n = 14), and ST636 (n = 10). Obtained results evaluated that circulating CRAB clones in Serbia were as follows:bla(OXA66)/bla(OXA23)/ST2 (32.4%),bla(OXA66)/bla(OXA23)/bla(OXA72)/ST2 (2.7%),bla(OXA66)/bla(OXA72)/ST492 (37.8%), andbla(OXA66)/bla(OXA72)/ST636 (27.1%). Conclusion This study revealed extremely high proportions of carbapenem resistance amongA. baumanniiclinical isolates due to the emergence ofbla(OXA-72),bla(OXA-23), andbla(NDM-1)genes among CRAB isolates in Serbia and their clonal propagation
GABA potentiate the immunoregulatory effects of Lactobacillus brevis BGZLS10-17 via ATG5-dependent autophagy in vitro
Get PDFThe characterization of mechanisms involved in the positive effects of probiotic bacteria in various pathophysiological conditions is a prerogative for their safe and efficient application in biomedicine. We have investigated the immunological effects of live bacteria-free supernatant collected from GABA-producing Lactobacillus brevis BGZLS10-17 on Concanavalin A-stimulated mesenteric lymph node cells (MLNC), an in vitro model of activated immune cells. We have shown that GABA containing and GABA-free supernatant of Lactobacillus brevis BGZLS10-17 have strong immunoregulatory effects on MLNC. Further, GABA produced by this strain exhibit additional inhibitory effects on proliferation, IFN-gamma and IL-17 production by MLNC, and the expression of MHCII and CD80 on antigen presenting cells. At the other hand, GABA-containing supernatants displayed the strongest stimulatory effects on the expression of immunoregulatory molecules, such as Foxp3(+), IL-10, TGF-beta, CTLA4 and SIRP-alpha. By looking for the mechanisms of actions, we found that supernatants produced by BGZLS10-17 induce autophagy in different MLNC, such as CD4(+) and CD8(+) T lymphocytes, NK and NKT cells, as well as antigen presenting cells. Further, we showed that the stimulation of Foxp3(+), IL-10 and TGF-beta expression by BGZLS10-17 produced GABA is completely mediated by the induction of ATG5 dependent autophagy, and that other molecules in the supernatants display GABA-, ATG5-, Foxp3(+)-, IL-10- and TGF-beta- independent, immunoregulatory effects
Evaluation of global haemostatic assays and fibrin structure in patients with pre-eclampsia
No full textIntroduction Haemostatic balance shifted towards hypercoagulability in normal pregnancy is even more pronounced in pre-eclampsia (P-EC). The aim of this study was to analyse haemostatic disturbances and fibrin clot properties in women with pre-eclampsia and to investigate their association with maternal and foetal outcomes. Methods Forty-six pregnant women diagnosed with pre-eclampsia were included in the study, with blood sampling done on the morning following admission to hospital, as well as after delivery (mean duration 4.8 days). Two global haemostatic assays-endogenous thrombin potential (ETP) and assay of overall haemostatic potential (OHP)-were employed, including fibrin clot turbidity measurements and scanning electron microscopy (SEM) of representative samples. Results Three thrombin generation parameters (ETP, t_lag and peak height) and OHP were significantly increased in pre-eclampsia compared with controls, whereas overall fibrinolytic potential (OFP-determined as a parameter of the OHP assay) had significantly lower values. Clot lysis time was significantly prolonged in patients with pre-eclampsia. In the pre-eclamptic group after delivery, we observed a significant elevation in the peak height and a reduction in the time to peak and OFP compared with values before delivery. Pre-eclamptic patients with renal complications had significantly higher values for ETP, peak height and D-dimer. Turbidity measurements and SEM revealed dense fibrin structure in patients with pre-eclampsia. Conclusion Patients with pre-eclampsia have enhanced coagulation and impaired fibrinolysis before, and even after, delivery. In particular, the presence of multi-organ dysfunction, such as renal dysfunction, may be associated with increased thrombin generation in pre-eclampsia
Thermal properties of 3-hydroxy fatty acids and their binary mixtures as phase change energy storage materials
No full textIn the present work, we describe the chemical synthesis of 3-HFAs as prominent derivatives of fatty acids and assess if they could be applied as phase change materials (PCM). In addition, 3-HFAs were obtained by depolymerization of a bacterial biopolymeric material, polyhydroxyalkanoate. Thermal properties of 3-hydoxyoctanoic, decanoic, and dodecanoic acids are reported for the first time. These materials showed the potential to be applied as PCM in temperature range from 33 degrees C to 66 degrees C. In order to expand the temperature range for application of 3-HFAs as PCM, eutectic mass ratios of three kinds of binary mixtures of 3-HFAs were calculated, and their properties were predicted using the Schroder-van Laar equation. Thermal properties of these mixtures were validated by differential scanning calorimetry (DSC) analysis. These results showed that eutectics considerably expanded the scope of applications of 3-HFAs as PCMs. 3-HFAs originating from biotechnologically obtained polyhydroxyalkanoates also showed potential to be applied in development of PCMs
Chapter 8 - Human genetic diversity in health and disease
No full textEstimated human genetic diversity is 0.1%, meaning that any two individuals differ at approximately one in every thousand base pairs. A small proportion of these genetic variants have functional consequences affecting health, including susceptibility to infections, diseases, and response to different drugs. Advancements in high-throughput sequencing technology have provided more insight into human genetic makeup and enabled better understanding of the link between genetic variations and health outcomes. However, despite considerable progress in human genetics, there are still many diseases that, given their complexity, cannot be explained simply by the genetic variants present and the involvement of factors such as epigenetics, environmental factors, and gene–environment interactions must be considered. We will discuss how human genetic diversity helps us to uncover the underlining mechanisms in monogenic, complex, and infectious diseases leading to a personalized approach to drug development and treatment
Diversity of non-starter lactic acid bacteria in autochthonous dairy products from Western Balkan Countries - Technological and probiotic properties
No full textThe aim of this review was to summarize the data regarding diversity of non-starter lactic acid bacteria (NSLAB) isolated from various artisanal dairy products manufactured in Western Balkan Countries. The dairy products examined were manufactured from raw cow's, sheep's or goat's milk or mixed milk, in the traditional way without the addition of commercial starter cultures. Dairy products such as white brined cheese, fresh cheese, hard cheese, yogurt, sour cream and kajmak were sampled in the households of Serbia, Croatia, Slovenia, Bosnia and Herzegovina, Montenegro, and North Macedonia. It has been established that the diversity of lactic acid bacteria (LAB) from raw milk artisanal dairy products is extensive. In the reviewed literature, 28 LAB species and a large number of strains belonging to the Lactobacillus, Lactococcus, Enterococcus, Streptococcus, Pediococcus, Leuconostoc and Weissella genera were isolated from various dairy products. Over 3000 LAB strains were obtained and characterized for their technological and probiotic properties including: acidification and coagulation of milk, production of aromatic compounds, proteolytic activity, bacteriocins production and competitive exclusion of pathogens, production of exopolysaccharides, aggregation ability and immunomodulatory effect. Results show that many of the isolated NSLAB strains had one, two or more of the properties mentioned. The data presented emphasize the importance of artisanal products as a valuable source of NSLAB with unique technological and probiotic features important both as a base for scientific research as well as for designing novel starter cultures for functional dairy food.This is the peer-reviewed version of the paper: (1) Terzić-Vidojević, A.; Veljović, K.; Tolinački, M.; Živković, M.; Lukić, J.; Lozo, J.; Fira, Đ.; Jovčić, B.; Strahinić, I.; Begović, J.; Popović, N.; Miljković, M.; Kojić, M.; Topisirović, L.; Golić, N. Diversity of Non-Starter Lactic Acid Bacteria in Autochthonous Dairy Products from Western Balkan Countries - Technological and Probiotic Properties. Food Research International 2020, 136. [https://doi.org/10.1016/j.foodres.2020.109494].Published version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1353
Bio Innovation of a Circular Economy for Plastics (BioICEP)
Get PDFProject ‘EcoPlastiC’ is putting PET plastics into a perpetual bio-cyclable loop. Polyethylene terephthalate (PET), a polymer used extensively in single-use packaging and beverage bottles as well as in the textiles industry, is not very circular. There is a growing need to convert unrecyclable post-use PET into new, high-performance bioplastics. In this case, post-use materials become ingredients for new products. The EU-funded EcoPlastiC project will optimise PET circularity. It will convert lower-grade PET and mixed recalcitrant PET plastic waste into high-performing biopolymers, through the development of a suite of breakthrough technologies adaptable to the waste input. For instance, it will develop a series of mechano-green, chemical and biocatalytic technologies to depolymerise PET. Moreover, microbiome processing will be used to produce new biopolymers.Principal Investigator: Dr Margaret Fournet, Athlone Institute of Technology, IrelandCoordinator for IMGGE: Dr Jasmina Nikodinović-RunićDuration period: 2020-202
Coding and noncoding variants of disease-causing genes of maturity-onset diabetes of the young: phenotype modulators and regulators of gene expression
Get PDFDijabetes adultnog tipa kod mladih (MODY) je nasledni, autozomno dominantni oblik dijabetesa, klinički i genetički veoma heterogen, koji nastaje kao posledica primarne disfunkcije β-ćelija pankreasa. MODY nastaje usled genetičkih varijanti u jednom od brojnih gena asociranih sa MODY dijabetesom, te je metodom sekvenciranja nove generacije i metodom istovremenog umnožavanja vezanih proba, prvi put u Srbij, analizirano 13 gena uzročnika MODY dijabetesa kod 29 klinički suspektnih pedijatrijskih pacijenata. Kombinovanjem ove dve metode detektovano je 20 različitih varijanti kod 75,9% pacijenata u 4 različita gena. Varijante u dva gena, GCK i HNF1B, detekotvane redom, kod 50% i 22,7% pacijenata, predstavljale su glavne uzročnike MODY dijabetesa u ovoj grupi pacijenata. U ovoj studiji identifikovana je i jedna nova prethodno neprijavljena varijanta u GCK genu, c.596T>C; p.Val199Ala, za koju su in silico predikcije nedvosmisleno pokazale da je patogena. Kako bi se detektovale varijante u promotorskom regionu, koje takođe dovode do MODY dijabetesa, analizirani su promotorski regioni četiri najčešća gena uzročnika MODY dijabetesa. Varijantni set u promotrskom regionu GCK gena (−282C>T; −194A>G; 402C>G) i varijanta c.-154-160insTGGGGGT u promotoru HNF1A gena, odabrane su kako bi se u funkcionalnim esejima u ćelijskoj kulturi ispitao njihov uticaj na ekspresiju datih gena. Analizirane su i interakcije ovih potencijalnih regulatornih elemenata sa transkripcionim faktorima u esejima usporene elektroforetske pokretljivosti. Rezultati funkcionalne analize odabarnih promotorskih varijanti ukazali su da varijante u promotoru, osim potencijalno patogenog efekta, mogu da imaju i ulogu modifikatora fenotipa, čime su dopunjena postojeća znanja o varijantama u promotorima MODY gena.Maturity-onset diabetes of the young (MODY) is an inherited, autosomal dominant form of diabetes, clinically and genetically very heterogeneous, resulting from primary β-cell dysfunction. Since MODY diabetes is caused by single gene variants in one of the many MODY-related genes, next generation sequencing and multiplex ligation-dependent probe amplification analysis were used to analyze 13 MODY-relate genes in 29 clinically suspected pediatric patients, for the first time in Serbia. Combining these two methods, 20 different variants, found in 4 genes, were identified in 75.9% patients. Variants in the GCK and HNF1B gene, detected in 50% and 22.7% patients, respectively, were the main cause of MODY diabetes in our patients. Also, a novel variant in the GCK gene: c.596T>C, p.Val199Ala was identified and predicted to be pathogenic by in silico algorithms. Due to the fact that promoter variants can also lead to MODY diabetes, promoter regions of the four most common MODY genes were screened. Two variants, the variant set in the promoter region of the GCK gene (−282C>T; −194A>G; 402C>G), and the variant c.-154-160insTGGGGGT in the promoter of the HNF1A gene, were selected to analyze their effect on gene expression in functional cell culture studies. Also, electrophoretic mobility shift assay was carried out to investigate the interaction of potential transcription factors with the promoter region surrounding the variant, as well as whether the variants affect the binding of those transcription factors. The results of the functional analysis of the selected promoter variants indicated that variants in the promoter, in addition to potentially pathogenic effect, may also play the role of a phenotype modifier, thus supplementing the existing knowledge about variants in the promoters of MODY genes
