imagine (Institute of molecular genetics and genetic engineering)
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GABA-Producing Natural Dairy Isolate From Artisanal Zlatar Cheese Attenuates Gut Inflammation and Strengthens Gut Epithelial Barrier in vitro
Get PDFProbiotic bacteria are recognized for their health-promoting properties, including maintenance of gut epithelial integrity and host immune system homeostasis. Taking into account the beneficial health-promoting effects of GABA, the presence of the gadB gene, encoding glutamate decarboxylase that converts L-glutamate to GABA, was analyzed in Lactic Acid Bacteria (LAB) natural isolates from Zlatar cheese. The results revealed that 52% of tested Lactobacillus spp. and 8% of Lactococcus spp. isolates harbor the gadB gene. Qualitative and quantitative analysis of GABA production performed by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) revealed the highest GABA production by Lactobacillus brevis BGZLS10-17. Since high GABA-producing LAB natural isolates are the most valuable source of naturally produced GABA, the probiotic properties of BGZLS10-17 were characterized. This study demonstrated high adhesion of BGZLS10-17 strain to Caco2 cells and the ability to decrease the adhesion of Escherichia coli ATCC25922 and Salmonella enterica C29039. Treatment of differentiated Caco-2 cells monolayer with BGZLS10-17 supernatant containing GABA alleviated inflammation (production of IL-8) caused by IL-1 beta and significantly stimulated the expression of tight junction proteins (zonulin, occludin, and claudin 4), as well as the expression of TGF-b cytokine leading to the conclusion that immunosuppression and strengthening the tight junctions can have significant role in the maintenance of intestinal epithelial barrier integrity. Taken together the results obtained in this study support the idea that using of GABA producing BGZLS10-17 probiotic strain could be a good strategy to modulate immunological response in various inflammatory diseases, and at the same time, it could be a good candidate for adjunct starter culture for production of GABA-enriched dairy foods and beverages offering new perspectives in designing the novel functional foods
In vitro digestion of meat- and cereal-based food matrix enriched with grape extracts: How are polyphenol composition, bioaccessibility and antioxidant activity affected?
Get PDFThe aim of this study was to evaluate the effect of enriching a complex food matrix (FM) with grape extracts on polyphenol content, composition, bioaccessibility and antioxidant activity during digestion. The grape extracts and FM were separately tested under the same conditions as controls. The FM by itself contains a significant amount of phenolic acids and flavonols, influencing the final recovery of polyphenols from grape extracts. The FM significantly increased the total recovery of polyphenols after digestion of grape seed extracts compared to those digested without the FM; however, a low recovery of proantocyanidins and total flavonoids was observed. Digestive fluids and FM compounds significantly increased the total polyphenol content of grape digests and significantly contributed to their ABTS(center dot+) scavenging activity and ferrous-ion-chelating capacity. The present study suggested that enrichment of meat-and cereal-based products with grape polyphenol extracts could be a good strategy to formulate a healthier diet.This is the peer reviewed version of the paper: Pešić, M. B., Milinčić, D. D., Kostić, A. Ž., Stanisavljević, N. S., Vukotić, G. N., Kojić, M. O., Gašić, U. M., Barać, M. B., Stanojević, S. P., Popović, D. A., Banjac, N. R., & Tešić, Ž. Lj. (2019). In vitro digestion of meat- and cereal-based food matrix enriched with grape extracts: How are polyphenol composition, bioaccessibility and antioxidant activity affected? Food Chemistry, 284, 28–44. [https://doi.org/10.1016/j.foodchem.2019.01.107][https://www.sciencedirect.com/science/article/abs/pii/S030881461930175X?via%3Dihub
Antimicrobial activity of novel chalcones and modulation of virulence factors in hospital strains of Acinetobacter baumannii and Pseudomonas aeruginosa
No full textAcinetobacter 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
Analysis of the role of SOX1 and SOX3 genes in promotion of malignant phenotype of glioblastoma cells
No full textGlioblastom, glioma tumor gradusa IV, je najčešći maligni tumor mozga kododraslih i jedan od najsmrtonosnijih tipova tumora. I pored agresivne terapije koja obuhvatahirurško uklanjanje tumora, radio- i hemio-terapiju, prosečno preživljavanje bolesnika saovim tipom tumora je oko 15 meseci. Glioblastom (GBM), pored tumorskih ćelija, sadrži ipopulaciju samo-obnavljajućih tumor-inicirajućih matičnih ćelija (matične ćelijeglioblastoma) koje se smatraju odgovornim za nastanak, progresiju, metastaziranje irezistenciju na terapiju.Geni SOXB1 podgrupe (SOX1, SOX2 i SOX3) kodiraju regulatorne proteine kojiimaju značajne uloge u mnogim procesima u toku razvića, kao što su održavanjepluripotentnosti matičnih ćelija i održavanje populacije neuralnih progenitora upluripotentnom i proliferišućem stanju. Ovi geni imaju i značajne funkcije u procesukarcinogeneze. Ekspresija gena ove podgrupe detektovana je u GBM. Funkcija gena SOX2je dobro proučena kod ovog tipa tumora; pokazano je da ovaj gen promoviše malignipotencijal ćelija GBM i neophodan je za održavanje tumorogenog potencijala matičnihćelija glioblastoma. Za razliku od gena SOX2, uloga gena SOX1 i SOX3 u ćelijama GBMjoš uvek nije dovoljno istražena. Stoga, u okviru ove doktorske disertacije analizirana jeuloga ovih gena u ćelijama glioblastoma.Dobijeni rezultati pokazuju da ćelijske linije GBM eksprimiraju gen SOX1. Poredtoga, u uslovima utišane ekspresije proteina SOX1 detektovano je smanjenjeproliferativnog kapaciteta, vijabilnosti i migratornog potencijala U251 ćelija GBM, kao ipovećanje broja ovih ćelija u senescenciji. Nakon dediferencijacije ćelijskih linija GBMdetektovano je povećanje ekspresije gena SOX1 u poređenju sa ekspresijom uočenom unjihovim parentalnim ćelijama. Nivo ekspresije gena SOX1 povećan je u kulturamamatičnih ćelija glioblastoma u poređenju sa ekspresijom ovog gena u imortalizovanim U87i U251 ćelijama; pri diferencijaciji ovih kultura uočeno je smanjenje ekspresije gena SOX1.Utišavanje ekspresije gena SOX1 u GNS166 kulturi matičnih ćelija GBM dovodi dosmanjenja proliferativnog kapaciteta i vijabilnosti ovih ćelija...one of deadliest cancers. Despite aggressive treatment, including surgical resection,chemotherapy and radiation, the median survival of patients with glioblastoma is 15months. A growing body of evidence indicates that GBM contains a population of selfrenewingtumor-initiating cells (glioblastoma stem cells - GSCs) that drive tumor initiation,propagation, metastasis and therapy resistance.SOXB1 genes (SOX1, SOX2 and SOX3) encode transcription regulators withimportant roles in embryonic development and carcinogenesis. Literature date revealed thatSOXB1 genes are expressed in GMB tumor samples. The role of one member of this group,SOX2 gene, is well documented in GBM. It was shown that SOX2 gene promotes malignantpotential of GBM cells and it is mandatory for maintenance of tumorogenicity of GSCs.Since the function of SOX1 and SOX3 genes in GBM still remains to be established, theaim of this thesis was to analyze the role of these genes in GBM.Obtained results demonstrated that all analyzed GBM cell lines express SOX1.Downregulation of this gene expression decreases proliferation, viability and migration,and induces senescence of U251 cells. Furthermore, dedifferentiation of GBM cells isaccompanied by increase of SOX1 level compared to that in parental cells. Expression ofthis gene was significantly increased in patient-derived GSC cultures compared to that inU87 and U251 cells; downregulation of SOX1 gene expression was seen upondifferentiation of GSCs. In addition, knock-down of this gene expression reducedproliferation and viability of GSCs.Results obtained in this thesis reveal SOX3 expression in GBM cell lines and tumortissue; the expression of this gene was elevated in the most of analyzed GBM samplescompared to expression levels detected in non-tumoral brain tissues. A high SOX3expression was not associated with the overall survival of GBM patients. Ectopicoverexpression of this gene increased proliferation, viability, migration and invasion ofGBM cells..
Frequency of FV Leiden and FII G20210A Mutations in Patients with Inherited Antithrombin Deficiency from Serbia
Get PDFPURPOSE: Thrombosis is multicausal disease in which both acquired and genetic risk factors play
important roles. The most frequent genetic risk factors known to date are the Factor V G1691A (FV
Leiden) and FII G20210A mutations. On the other hand, inherited antithrombin (AT) deficiency, caused
by mutations in the AT gene (SERPINC1) is a very rare disorder, but it is associated with significant risk
for thrombotic complications. AT deficiency is classified into two types: type-I is a quantitative disorder
characterized by decreased amount and activity of AT, while type-II is a qualitative - functional disorder.
Aim of our study was to analyze the frequency of FV Leiden and FII G20210A mutations in patients with
inherited AT deficiency from Serbia.
METHODOLOGY: A study was carried out in large group of AT deficiency patients from Serbia. Cohort
of 42 subjects (15m/27f; 36.7±18.7y) from 18 Serbian families included 24 symptomatic and 18
asymptomatic first-degree relatives. Among them, type-I AT deficiency were detected in 9 families (19
members: 6m/13f; 37.1±19.0y) and type-II in 9 families (23 members: 9m/14f; 36.5±18.8y). FV Leiden
and FII G2010A mutations were detected by PCR, followed by digestion with specific restriction enzymes
(PCR-RFLP).
RESULTS: We have detected 3 FV Leiden heterozygous carriers in 3 different families (1 with type-I
and 2 with type-II AT deficiency). All 3 carriers were symptomatic. Regarding FII G20210A mutation, 2
heterozygous carriers, both asymptomatic and from same family with type-I deficiency, were identified.
According to our findings in families with AT deficiency from Serbia frequency of FV Leiden and FII
G20210A mutation are 16.7% and 5.6%, respectively.
CONCLUSION: This is the first study in which frequency of FV Leiden and FII G20210A mutations in
patients with inherited AT deficiency from Serbia were examined. Results of our study suggest that
these mutations can be relevant for AT deficiency patients’ phenotype, but further studies are required.Synergy in Hemophilia; 16th International Hemophilia Congress of Turkey, April 2019, Antaly
Tailoring the physico-chemical and antimicrobial properties of agar-based films by in situ formation of Cu-mineral phase
No full textNew agar-based composite films with increasing Cu-carbonate and Cu-phosphate mineral phase content were prepared by in situ mineralization and solvent casting method. SEM and optical analysis revealed that Cu-carbonate phase had better compatibility with agar matrix than Cu-phosphate phase. Incorporation of both mineral phases improved mechanical and water vapor barrier properties of the obtained mineralized films, in concentration dependent manner. When 5 mM of carbonate precursor was incorporated into agar matrix, mechanical resistance was enchanced for 44% and water vapor barrier property for 40%. The release of Cu (II) was higher in acidic conditions for both mineralized composites and remained in the range of specific release limits for this metal. In addition, both mineralized composite films exhibited distinctive antimicrobial activity against Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria. Overall, the Cu-carbonate and Cu-phosphate mineralized agar films showed potential to be used for food packaging materials, agriculture or medical purposes
Genes and metabolic pathway of sarcoidosis: identification of key players and risk modifiers
Get PDFIntroduction: Sarcoidosis is a rare multisystem granulomatous disease with unknown etiology. The interplay of vitamin D deficiency and genetic polymorphisms in genes coding for the proteins relevant for metabolism of vitamin D is an important, but unexplored area. The aim of this study was to investigate the association between single nucleotide polymorphisms (SNPs) in CYP2R1 (rs10741657), CYP27B1 (rs10877012), DBP (rs7041; rs4588), and VDR (rs2228570) genes and sarcoidosis, as well as the association between these SNPs and 25(OH)D levels in sarcoidosis patients. Material and methods: For that purpose we genotyped 86 sarcoidosis patients and 50 healthy controls using the PCR-RFLP method. Results: Subjects carrying the CC genotype of CYP27B1 rs10877012 have 10 times lower odds of suffering from sarcoidosis. Moreover, DBP rs4588 AA genotype was shown to be a susceptibility factor, where carriers of this genotype had eight times higher odds for developing sarcoidosis. In addition, the A allele of the DBP gene (rs4588) was associated with lower levels of 25(OH)D in sarcoidosis patients. Conclusions: These results suggest that patients with vitamin D deficiency should be regularly tested for genetic modifiers that are related to sarcoidosis in order to prevent development of serious forms of sarcoidosis
Genomic data provide new insights on the demographic history and the extent of recent material transfers in Norway spruce
Get PDFPrimeval forests are today exceedingly rare in Europe, and transfer of forest reproductive material for afforestation and improvement has been very common, especially over the last two centuries. This can be a serious impediment when inferring past population movements in response to past climate changes such as the last glacial maximum (LGM), some 18,000 years ago. In the present study, we genotyped 1,672 individuals from three Picea species (P. abies, P. obovata, and P. omorika) at 400K SNPs using exome capture to infer the past demographic history of Norway spruce (P. abies) and estimate the amount of recent introduction used to establish the Norway spruce breeding program in southern Sweden. Most of these trees belong to P. abies and originate from the base populations of the Swedish breeding program. Others originate from populations across the natural ranges of the three species. Of the 1,499 individuals stemming from the breeding program, a large proportion corresponds to recent introductions from mainland Europe. The split of P. omorika occurred 23 million years ago (mya), while the divergence between P. obovata and P. abies began 17.6 mya. Demographic inferences retrieved the same main clusters within P. abies than previous studies, that is, a vast northern domain ranging from Norway to central Russia, where the species is progressively replaced by Siberian spruce (P. obovata) and two smaller domains, an Alpine domain and a Carpathian one, but also revealed further subdivision and gene flow among clusters. The three main domains divergence was ancient (15 mya), and all three went through a bottleneck corresponding to the LGM. Approximately 17% of P. abies Nordic domain migrated from P. obovata ~103K years ago, when both species had much larger effective population sizes. Our analysis of genomewide polymorphism data thus revealed the complex demographic history of Picea genus in Western Europe and highlighted the importance of material transfer in Swedish breeding program
Current state and prospects of biotechnology in Central and Eastern European countries. Part II: new and preaccession EU countries(CRO, RO, B&H, SRB)
No full textInnovation holds the potential for economic prosperity. Biotechnology (BT) has proved to be a viable vehicle for the development and utilization of technologies, which has brought not only advances to society, but also career opportunities to nation-states that have enabling conditions. In this review, we assess the current state of BT-related activities within selected new and preaccession EU countries (NPA) of CEE region namely Croatia, Romania, Bosnia and Herzegovina and Serbia, examining educational programs, research activity, enterprises, and the financing systems. The field of BT covers a broad area of activities, including medical, food and agriculture, aquaculture or marine, environmental, biofuels, bioinformatics, and many others. Under the European Commission (EC), member-states are to set their Research and Innovation Strategies for Smart Specialization (RIS3), to identify priorities or strengths in order to develop knowledge intensive economies. As the four countries highlighted in this review are in the early stages of implementing RIS3 or have not yet fully formulated, it presents an opportunity to learn from the successes and failures of those that have already received major structural funds from the EC. A critical point will be the ability of the public and private sectors' actors to align, in the implementation of RIS3 as new investment instruments emerge, and to concentrate efforts on a few select target goals, rather than distribute funding widely without respect to a long-term vision
