imagine (Institute of molecular genetics and genetic engineering)
Not a member yet
3088 research outputs found
Sort by
Stability and bioactive compounds assessment of yogurt containing novel natural starter cultures with the ability to promote longevity in Caenorhabditis elegans
Get PDFYogurt 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
Expression profile of CD81 gene transcripts in colorectal cancer
Get PDFThe activity profi le of alternative promoters may be an indicator of tumor characteristics. Alternative promoters of CD81 gene were shown to be differentially active in colon and rectal cancer tissue. The promoter active in colon and rectal cancer gives rise to transcripts CD81-205 and CD81-215, while the promoter active in normal gut mucosa gives rise to transcripts CD81-203 and CD81-213. This study aimed to analyze the relative abundance of the CD81 gene transcripts in colorectal cancer
Biocorrosion, biofouling and health risk: biological activity reaction tests of selected brackish groundwater occurrences in Serbia
Get PDFTargeted physiological groups of bacteria were cultivated and identified in the brackish
groundwaters of Obrenovačka Banja (OB), Lomnički Kiseljak (LK) and Velika Vrbnica (VV) using biological
activity reaction tests (BARTs) to assess the biocorrosion, biofouling and health risks. The highest density of
iron-related, sulfate-reducing, slime-forming, facultatively anaerobic heterotrophic, denitrifying bacteria and
representatives of Pseudomonas spp. was recorded in the OB sample, while the lowest density of the same
physiological groups of bacteria was recorded in the LK sample. Facultatively anaerobic heterotrophic
bacteria were the most abundant in the OB and LK samples, while, in contrast, heterotrophic aerobic bacteria
were the most abundant in the VV sample. All tested samples were characterized by a high degree of
biochemical activity associated with iron-related, sulfate-reducing, slime-forming, heterotrophic aerobic and
facultatively anaerobic bacteria. Also, high biochemical activity of denitrifying bacteria was recorded in the
OB sample, and the same activity of Pseudomonas species was recorded in the OB and VV samples. For OB
and LK groundwaters, the highest degree of risk was estimated for biocorrosion process, while for the OB
and VV occurrences, the highest degree of risk was estimated for biofouling process. The health risk was
present for all examined groundwaters. Caution is warranted in further use of all investigated occurrences
due to the established public health risk and an immediate revitalization of the OB, LK and VV wells is
necessary.2nd International Conference on Chemo and BioInformatics, ICCBIKG 2023, September 28-29, 2023 Kragujevac,
Serbi
Novel aryl hydrocarbon receptor modulator promotes immunosupressive immune response by stimulating T regulatory cells in the gut
Get PDFIntroduction: The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor which is highly expressed in mucosal tissues - by epithelial cells and immune cells such as Th17 CD4+ and T reg- ulatory cells (Treg). Besides its function of clearing environmental pollutants from the body, it was also revealed that AhR has immunoregulatory effects, thus becoming a potential therapeutic target for mod- ulating the immune response. For that purpose we tested a novel synthetic AhR modulator under the code name C43.
Methods: CYP1A1 (downstream effector of AhR) activation was tested by the EROD assay. Sort-purified CD4+ cells from mesenteric lymph nodes (MLN) were treated with C43 for 24 h. Zebrafish embryos were used to test the toxicity of C43. Male C57BL/6 mice orally received C43 (10 mg/kg) for 5 consecutive days, after which MLN were harvested. Phenotype and function of the cells were analyzed by flow cytometry. Results: C43 showed mild AhR agonistic activity. After treating the sort-purified CD4+ cells with C43, there was a shift in the Th17/Treg ratio in favour of the latter. C43 showed no signs of toxicity when tested on zebrafish embryos. MLN cells from mice that received C43 revealed a shift in the Th1/Treg ratio in favour of Tregs, with a documented rise of the portion of Tregs that expressed CYP1A1 in comparison with the control group of mice.
Conclusion: C43 can modulate the immune response through the intestine by promoting the im- munosuppressive Treg population
Influence of amino acid substitution on the antimicrobial activity of bacteriocin lactolisterin BU
Get PDFIntroduction: Lactolisterin BU (LBU) is a potent bacteriocin derived from Lactococcuslactis subsp. lactisbv. diacetylactis BGBU1-4. It exhibits antimicrobial properties against Gram-positive food spoilage andfoodborne pathogens. This research aimed to explore the impact of amino acid substitution in LBU onits antimicrobial activity by utilizing in silico prediction of LBU’ssecondary structure and amino acid substitutions.Methods: The secondary structure of LBU was predicted using Phyre2 software. Five variants of LBUwere selected and chemically synthesized, along with unaltered LBU and BHT-B,serving as controls. Peptides were twofold diluted in distilled water, resulting in final concentrations ranging from 1000 µg/mlto 0.5 µg/ml. An agarspot test, employing 5 µl of the dilution, was conducted on three indicatorstrains:Lactococcus lactis BGMN1-596, Listeria monocytogenes ATCC19111, and Staphylococcus aureusATCC25923. The presence of inhibition zones was analyzed after overnight incubation at 37°C (S. aureus)and 30°C (L. lactis and L. monocytogenes).Results: Phyre2 analysis unveiled the presence of two α-helices in LBU’s structure. The majority of LBUvariants displayed altered antimicrobial activity, with some changes being genusspecific, potentially attributable to variances in cell wall composition. Some variants completely lost their activity, underscoring the significance of native amino acids or their physicochemical properties in the correspondingpositions within LBU’s structure. Furthermore, it was confirmed that chemically synthesized LBU effectively retains its antimicrobial activity.Conclusion: Changesin amino acid composition give insight on structure-function relationship of LBU
Drying without dying: revealing the role of late embryogenesis abundant proteins during desiccation in Ramonda serbica
Get PDFIntroduction: Resurrection plants (such as Ramonda serbica) can survive a long desiccation period and
fully resume their metabolism upon watering. The hallmark of desiccation tolerance (DT) is the accumulation of protective, intrinsically disordered proteins(IDPs), called late embryogenesis abundant proteins (LEAPs). Although their high structural plasticity allows them to interact with various partners, no
specific cellular targets of LEAPs have been identified so far.
Methods: To identify LEAPsinvolved in DT, differential transcriptome and proteome analyses of hydrated
and desiccated R. serbica leaves were performed. The identified LEAPs were structurally characterised
and classified. To evaluate theirstructural propertiesin vitro and their potential functionsin vivo, the representative RsLEA proteins, were produced in Escherichia coli using recombinant DNA technology.
Results: Members of the LEA4 protein family represent the majority of desiccation-inducible LEAPs. Even
17 proteins belonging to the LEA4 protein family group were induced by desiccation. They show high disorder propensity (82 %), and at the same time, a high tendency to form α-helices (>80%). Although recombinant DNA technology has traditionally been used to overexpress and purify various globular
proteins, the production of IDPsis challenging due to their high susceptibility to proteolytic cleavage and
aggregation. Nevertheless, the representative LEAPs containing hexa-Histagsimmunoglobulin G-binding protein and a proteolytic TEV site were produced, purified and cleaved by TEV protease.
Conclusion: The combination of in silico and in vitro results will be crucial for the identification of endogenous partners of LEAPs, providing further insight into their role in DT
PB2037: NEW TERT VARIANT IN A FAMILY WITH APLASTIC ANEMIA
Get PDFBackground:TERT gene, the most frequently mutated gene in patients with telomere biology disorders (telomeropathies), encode telomerase reverse transcriptase enzyme. Heterozygous variants in the TERT gene impair telomerase activity by haploinsufficiency and pathogenic variants are associated with bone marrow failure syndrome and acute myeloid leukemia predisposition. TERT variants show incomplete penetrance and can also be found in asymptomatic family members. Some patients with telomeropathies present with severe symptoms at early age, and in other diseases may appear later in life like aplastic anemia, pulmonary or hepatic fibrosis. Affected families may show anticipation that may result in more severe forms of the disease in succeeding generations. Due to the rarity of the disease and the small number of clinical trials, telomeropathies are often unrecognized and misdiagnosed. Aims: To report a novel variant in TERT gene in familial hematopoietic disorder. Methods: Next Generation Sequencing of DNA isolated from peripheral blood of a patient (older sister) with clinical diagnosis of aplastic anemia, using TruSight One MiSeq platform (Illumina®) and segregation sequencing analysis of patient’s mother and younger sister. Results: We analyzed all three family members presented with a similar clinical appearance and hematology findings (moderate megaloblastic pancytopenia). Mother was diagnosed at age 14, but reevaluated before delivery in 2001 as hypoplastic MDS and trisomy 8 in karyotype with marked thrombocytopenia, being stable for years. Two daughters, both diagnosed as familiar aplastic anemia with normal karyotype, without elements of Fanconi anemia, at age of 13 and 14 yrs, also with profound thrombocytopenia without severe bleeding episodes. Moreover, lung and liver fibrosis were excluded. We identified a novel missense heterozygous variant c.2605G>A p.(Asp869Asn) in TERT gene in all three family members. This variant results in replacement of aspartic amino acid on 869 position in TERT enzyme polypeptide chain by asparagine. According to ACMG classification, detected variant is characterized as likely pathogenic, class 2. This variant is very rare and was detected in gnomAD exomes and gnomAD genomes data bases. It is located in highly conserved protein region and is very likely to disrupt the function of the enzyme. Summary/Conclusion: As patients with telomeropathies often have a history of macrocytosis and mild to moderate thrombocytopenia, that can be wrongly diagnosed as immune-mediated thrombocytopenia, myelodysplastic syndrome or moderate aplastic anemia, our findings indicate that TERT rare variants pass under-recognized in these patients. Therefore, this report emphasizes the importance for routine deep genetics screening for TERT rare variants in patients with family history of cytopenia, different bone marrow failure syndromes and aplastic anemia, regardless the age or clinical presentation. This investigation is able to identify clinically inapparent telomere biology disorder and improve outcomes through forehand diagnosis setting, genetic counseling and the precise therapy considerations especially stem cell grafting
22q11.2 microdeletion is the most common genomic abnormality in Serbian newborns with critical congenital heart disease and could be rapidly detected by Multiplex ligation probe amplification analysis
Get PDFBackground/Objectives: Genetic tests may facilitate rapid and
effective diagnostics but unfortunately their high costs usually
limit their application in all patients (1). We aimed to investigate
the utility of rapid, cost effective and high sensitive Multiplex
ligation probe amplification analysis (MLPA) for detection copy
number variants (CNV) in newborns with critical CHD, admitted to
the Neonatal Intensive Care Unit (NICU).
Methods: Study included 100 consecutive newborns admitted
to the NICU, University Children’s Hospital in Belgrade from
August 2014 to September 2019. Patients with viable trisomies
(21, 18 and 13) were excluded. All participants were tested by
MLPA analysis using SALSA MLPA P250-B2 Di George and SALSA
MLPA P311-B1 Congenital Heart Disease probemixes (MRC Holland,
The Netherland).
Results: Pathogenic CNVs were identified in ten (10%) patients.
Nine of them had 22q11.2 deletion detected by both kits while
one patient had 3p25 deletion detected by P311 kit.
Conclusion: Genetic evaluation of all newborns with critical
CHD admitted to the NICU by rapid and inexpensive MLPA analysis
using combination P250 and P311 SALSA probemixes could
contribute to high detection rate of pathogenic variants.Abstracts from the 55th European Society of Human Genetics
(ESHG) Conferenc
Analysis of cohort of patients with 22q11.2 deletion syndrome - a single-center experience from Serbia
Get PDFNeurodevelopmental disorders (NDDs), such as autism spectrum disorders (ASD),
intellectual disability (ID), schizophrenia, and bipolar disorder, are caused by
disruption of brain development. They affect approximately 4% of the European
population. However, molecular mechanisms underlying NDDs are still unknown.
One of the syndromes with a high risk for NDDs is 22q11.2 Deletion Syndrome
(22q11.2DS) caused by microdeletion 22q11.2. 22q11.2DS is the most common
microdeletion in humans; approximately, 25% of patients with 22q11.2DS develop
schizophrenia compared to 1% in the general population, while an ID is detected in
approximately 45% of patients and ASD in 14-50% of cases. We analyzed genomic
and clinical findings in our cohort of 35 patients with 22q11.2DS. The majority of
patients have 3 Mb deletion and nine patients have inherited 22q11.2 microdeletion
from their parents. Twenty-one different clinical presentations are revealed in the
cohort with developmental delay detected in about 50% of patients. Approximately
80% of patients have heart malformations, palatal clefts/velopharyngeal insufficiency
was detected in about 30% of them, facial dysmorphism in approximately 80% and
hypocalcemia was seen in about 20% of patients. Here we presented a cohort of
patients with 22q11.2DS which represents a good system for modeling NDDs in vitro.BOOK OF ABSTRACTS: 8th CONGRESS OF SERBIAN NEUROSCIENCE SOCIETY with international participation 31 May – 2 June 2023. Belgrade, Serbi
The role of specific SOX genes and microRNAs in reactivation and senescence of human astrocytes derived from pluripotent NT2/D1 cells
Get PDFAstrocytes are the main homeostatic cells in the brain with important roles both in
physiological and pathological conditions. They have a unique ability to become
reactivated in response to different types of brain pathologies, which serves as a
compensatory response that modulates tissue damage and recovery. Also, senescent
astrocytes have profound implications in age-related neurodegenerative disorders. The
molecular mechanisms underlying astrocyte reactivation and senescence are still not
well understood. To investigate the roles of SOX2 and SOX9 transcription factors and
miR-21 in these phenotypic alternations of astroglia, astrocytes derived from NT2/D1
cell line (NT2/A) were used as a model system. Western blot analyses showed that the
expression of both SOX2 and SOX9 decreases during the maturation of NT2/A and
they are re-expressed upon in vitro induced injury. Further modulation of the SOX2
and SOX9 expression will reveal their roles in the regulation of astrocytes
reactivation. Down-regulation of mir-21 in both immature and mature NT2/A by
using the antisense technology, induced the decline in cell proliferation revealed by
Ki67 proliferation marker. Also the premature cellular senescence was induced as
indicated by increase in SA-ß-gal activity and the expression of p21 and p53.
Additionally, in silico analysis predicted many of the genes, previously shown to be
upregulated in senescent astrocytes, as miR-21 targets.
Clarifying the roles of SOX genes and miRNAs in astrocyte reactivation and
senescence would contribute to better understanding of the functions of these cells at
the molecular level, which holds promise for development of new therapeutic
strategies.BOOK OF ABSTRACTS: 8th CONGRESS OF SERBIAN NEUROSCIENCE SOCIETY with international participation 31 May – 2 June 2023. Belgrade, Serbi