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Genomic and clinical findings in patients with 22q11.2 duplication syndrome
Get PDFNeurodevelopmental disorders (NDDs), such as autism spectrum disorders (ASD),
schizophrenia, and intellectual disability, are caused by disruption of early brain
development. NDDs represent important public health challenge in modern societies
with prevalence of about 10 to 15% of all births and the tendency of increasing
worldwide. On the other side, treatments of NDDs are focused on symptoms due to
limited understanding of underlying pathophysiological mechanisms. Individuals with
the 22q11.2 duplication syndrome (22q11.2dup), caused by heterozygous 22q11.2
microduplication, have an elevated risk of developing NDDs. Literature data revealed
that ASD is detected in 14-25% of patients with 22q11.2dup while schizophrenia is
less common in these patients than in the general population, suggesting that
22q11.2dup might be protective against schizophrenia. We investigated genomic and
clinical findings in cohort of 8 patients with 22q11.2dup. The majority of patients
have 3Mb duplication. Five patients have 22q11.2 microduplication inherited from
their parents. Other CNVs or SNVs are detected in 5 out of 8 patients. Common
medical anomalies in our cohort of patients include developmental delay, facial
dysmorphism, heart malformations, anomalies of the skeletal system, and anomalies
affecting the eye. Characterization of a cohort of patients with 22q11.2dup is
important since 22q11.2dup represents a powerful model to get insights into the
molecular mechanisms underlying NDDs.BOOK OF ABSTRACTS: 8th CONGRESS OF SERBIAN NEUROSCIENCE SOCIETY with international participation 31 May – 2 June 2023. Belgrade, Serbi
Two main skeletal muscle molecular phenotypes of mouse dm1 models: a comparative transcriptomic analysis
Get PDFIntroduction:Myotonic dystrophy type 1 (DM1) is a rare, incurable multisystemic disease, with the main
symptoms being skeletal muscle weakness, atrophy, and myotonia. It is caused by CTG expansion in the
3' UTR of the DMPK gene whose RNA acquires toxic functions and sequesters MBNL proteins, resulting
in globally altered RNA metabolism. To better understand the DM1 transcriptome, we systematically analyzed
gene expression in the skeletal muscles of various mouse DM1 models.
Methods:We retrieved 13 publicly available RNA-seq datasets from mouse models expressing expanded
CTG repeats (HSALR, CTG480KI, TREDT960I) and Mbnl knockout models (SKO, DKO, TKO). Our bioinformatic
pipeline with unified parameters consisted of preprocessing, differential expression (DESeq2),
gene network analysis (WGCNA), comparison of gene network interactions with the STRING database,
and network node enrichment analysis (Cytoscape).
Results: In models expressing CTG repeats, the average number of up-regulated genes was 787, compared
to 676 in Mbnl knockout models, while there was 642 and 380 down-regulated genes, respectively
(log2FC>1, padj>0.05). Both model groups had network modules whose nodes were enriched for muscle
and secretory functions (FDR<0.05). There were modules related to immune response, lipid transfer,
and insulin in models expressing repeats and modules related to immunoglobulins and extracellular
matrix in knockout models.
Conclusion: Gene expression patterns separated Mbnl knockouts from models expressing CTG repeats
that had a greater number of smaller functionally distinct network modules. Our results revealed novel
pathway changes in DM1 skeletal muscles, among which immunological and secretory are particularly
interesting as molecular targets for further investigation
Effect of prothrombin Belgrade mutation, causing antithrombin resistance, on fibrin clot properties
No full textIntroduction Prothrombin Belgrade mutation is the result of the c.1787G>A substitution in the prothrombin gene. It is located in the antithrombin and sodium binding site and leads to impaired inactivation of thrombin by antithrombin, resulting in antithrombin resistance and thrombotic disorders. However, it negatively affects sodium binding and may have hypocoagulant effects. Considering that prothrombin Belgrade mutation mechanism is still not fully elucidated and that sodium binding is important for thrombin affinity towards fibrinogen, our aim was to determine whether this mutation affects fibrin clot formation and lysis. Methods Using HEK293T cell line, recombinant wild type and mutated prothrombin were generated by transient transfection. Samples that correspond to plasma of a non-carrier, heterozygous and homozygous carriers were reconstituted using prothrombin deficient plasma and recombinant proteins. Reconstituted samples were used in OHP assay (Overall Hemostasis Potential) to determine kinetic profiles of coagulation and fibrinolysis. Clot turbidity assay was performed to observe kinetics of clot formation and lysis more closely. Fibrin clots formed in reconstituted plasma samples were analyzed by confocal microscopy to determine density of fibrin network. Fibrin clots were additionally observed using electron microscopy to determine thickness of individual fibrin fibers. Results No significant difference found in OHP, OCP, OFP, and fibrin network density between wild type, heterozygous, and homozygous carrier reconstituted plasma samples. There were significant differences between samples for slope and slope time parameters in kinetic profiles and fibrin fiber thickness. Conclusions Results indicate that prothrombin Belgrade mutation has no significant impact on fibrinolysis, however it may affect kinetics of clot formation and its architecture
Bioplastics upcycling loop (BioPolyCycle)
Get PDFBioPolyCycle aims to develop bio-upcycling technologies for current bioplastics to produce novel advanced bio-based and biodegradable materials for food packaging and other applications. Our upcycling approach encompasses utilization and conversion of commercially available bioplastic (e.g. currently marketed polylactide (PLA), polycaprolactone (PCL) and polyhydroxyalkanoates (PHA)) to carbon rich feedstocks to be used for biotechnological production of raw biopolymers (such as PHA and bacterial cellulose – BC). Simultaneously, efficient biocatalysts will be developed to be integrated into new biomaterials for increased degradability. Thus, bioplastics will be upcycled into new materials or products of better quality and better environmental value, ensuring that micro-plastics are avoided. This will also allow the sustainable recycling or biological degradation of produced materials.Principal Investigator: Dr Maciej GuzikCoordinator for IMGGE: Dr Jasmina Nikodinović-RunićDuration period: 2023-202
The production of Delftia biomass in continuous system growing on depolymerized PET
No full textThis dataset contains the process settings of the production of Delftia biomass on
depolymerized PET in a continuous system. While the microorganism exhibits rapid growth in batch
systems, the downtime required for batch reactor preparation decreases overall efficiency. To overcome
this limitation, a continuous system was evaluated using depolymerized PET. The obtained biomass was
assessed for its potential conversion into biodegradable films, as previous studies on biodegradable
films have indicated superior performance with protein-rich biomass compared to PHA-rich.
Consequently, the biomass production was optimized to enhance protein content.readme.txt (3.885Kb)***Dataset contents***
Batch phase 20231120.png (14.71Kb)
Process data 20231120.csv (2.371Mb)File readme.txt (3.885Kb) is under licence public domain CC
The impact of synbiotic treatment on the levels of gut-derived uremic zoxins, inflammation, and gut microbiome of chronic kidney disease patients - a randomized trial
Get PDFObjectiveAltering dysbiotic gut flora through synbiotic supplementation has recently been recognized as a potential treatment strategy to reduce the levels of gut-derived uremic toxins and decrease inflammation. Assessing its efficacy and safety has been the main goal of our randomized, double-blind, placebo-controlled study.MethodsA total of 34 nondialyzed chronic kidney disease patients, aged ≥18 years, with an estimated glomerular filtration rate between 15 and 45 mL/minute, were randomized either to an intervention group (n = 17), receiving synbiotic (Lactobacillus acidophilus, Lactobacillus casei, and Bifidobacterium lactis, 32 billion colony forming units per day plus 3.2 g of inulin), or control group (n = 17), receiving placebo during 12 weeks. The impact of treatment on the dynamic of serum levels of gut-derived uremic toxins, total serum indoxyl sulfate, p-cresyl sulfate, and trimethylamine N-oxide, was defined as the primary outcome of the study. Secondary outcomes included changes in the stool microbiome, serum interleukin-6 levels, high-sensitivity C-reactive protein, estimated glomerular filtration rate, albuminuria, diet, gastrointestinal symptom dynamics, and safety. Serum levels of uremic toxins were determined using ultraperformance liquid chromatography. The stool microbiome analysis was performed using the 16S ribosomal ribonucleic acid gene sequencing approach.ResultsSynbiotic treatment significantly modified gut microbiome with Bifidobacteria, Lactobacillus, and Subdoligranulum genera enrichment and consequently reduced serum level of indoxyl sulfate (ΔIS –21.5% vs. 5.3%, P < .001), improved estimated glomerular filtration rate (ΔeGFR 12% vs. 8%, P = .029), and decreased level of high-sensitivity C-reactive protein (–39.5 vs. –8.5%, P < .001) in treated patients. Two patients of the intervention arm complained of increased flatulence. No other safety issues were noted.ConclusionSynbiotics could be available, safe, and an effective therapeutic strategy we could use in daily practice in order to decrease levels of uremic toxins and microinflammation in chronic kidney disease patients
Manuka Honey/2-Hydroxyethyl Methacrylate/Gelatin Hybrid Hydrogel Scaffolds for Potential Tissue Regeneration
Get PDFScaffolding biomaterials are gaining great importance due to their beneficial properties for medical purposes. Targeted biomaterial engineering strategies through the synergy of different material types can be applied to design hybrid scaffolding biomaterials with advantageous properties for biomedical applications. In our research, a novel combination of the bioactive agent Manuka honey (MHo) with 2-hydroxyethyl methacrylate/gelatin (HG) hydrogel scaffolds was created as an efficient bioactive platform for biomedical applications. The effects of Manuka honey content on structural characteristics, porosity, swelling performance, in vitro degradation, and in vitro biocompatibility (fibroblast and keratinocyte cell lines) of hybrid hydrogel scaffolds were studied using Fourier transform infrared spectroscopy, the gravimetric method, and in vitro MTT biocompatibility assays. The engineered hybrid hydrogel scaffolds show advantageous properties, including porosity in the range of 71.25% to 90.09%, specific pH- and temperature-dependent swelling performance, and convenient absorption capacity. In vitro degradation studies showed scaffold degradability ranging from 6.27% to 27.18% for four weeks. In vitro biocompatibility assays on healthy human fibroblast (MRC5 cells) and keratinocyte (HaCaT cells) cell lines by MTT tests showed that cell viability depends on the Manuka honey content loaded in the HG hydrogel scaffolds. A sample containing the highest Manuka honey content (30%) exhibited the best biocompatible properties. The obtained results reveal that the synergy of the bioactive agent, Manuka honey, with 2-hydroxyethyl methacrylate/gelatin as hybrid hydrogel scaffolds has potential for biomedical purposes. By tuning the Manuka honey content in HG hydrogel scaffolds advantageous properties of hybrid scaffolds can be achieved for biomedical applications
Triggering and identifying the polyurethane and polyethylene-degrading machinery of filamentous fungi secretomes
No full textThe uncontrollable disposal of plastic waste has raised the concern of the scientific community, which tries to face this environmental burden by discovering and applying new techniques. Regarding the biotechnology field, several important microorganisms possessing the necessary enzymatic arsenal to utilize recalcitrant synthetic polymers as an energy source have been discovered. In the present study, we screened various fungi for their ability to degrade intact polymers, such as ether-based polyurethane (PU) and low-density polyethylene (LDPE). For this, ImpranIil® DLN-SD and a mixture of long-chain alkanes were used as sole carbon sources, indicating not only the most promising strains in agar plate screening but also inducing the secretion of depolymerizing enzymatic activities, useful for polymer degradation. The agar plate screening revealed three fungal strains belonging to Fusarium and Aspergillus genera, whose secretome was further studied for its ability to degrade the aforementioned non-treated polymers. Specifically for ether-based PU, the secretome of a Fusarium species reduced the sample mass and the average molecular weight of the polymer by 24.5 and 20.4%, respectively, while the secretome of an Aspergillus species caused changes in the molecular structure of LDPE, as evidenced by FTIR. The proteomics analysis revealed that the enzymatic activities induced in presence of Impranil® DLN-SD can be associated with urethane bond cleavage, a fact which was also supported by the observed degradation of the ether-based PU. Although, the mechanism of LDPE degradation was not completely elucidated, the presence of oxidative enzymes could be the main factor contributing to polymer modification.The authors would like to thank the VIB Proteomics Core for the contribution regarding the mass spectrometry-based proteomics experiments (EPIC-XS, project number 823839, funded by the Horizon 2020 programme of the European Union).Related to accepted version: [https://imagine.imgge.bg.ac.rs/handle/123456789/1800]Related to other materials: [https://zenodo.org/record/7767083#.ZCP8WXZBxPa
3D in vitro osteosarcoma model based on the biomimetic scaffolds and perfusion bioreactor supports in vivo-like osteosarcoma cell behaviour
No full textOsteosarcoma is a primary bone tumour that has a high tendency to metastasise on distant organs
with a devastating prognosis once this process occurs. Lack of significant breakthroughs in
osteosarcoma drug treatment together with the lasting scarcity of knowledge about the tumour itself
indicates the inherent weaknesses of dominantly used cell monolayers and animal models in drug
screening. 3D in vitro models emerged as an innovative counterpart to the traditional models
acknowledging the role of tumour cell microenvironment. Our 3D in vitro osteosarcoma model is
based on a bone-like macroporous alginate scaffold with incorporated hydroxyapatite and a perfusion
bioreactor in order to accurately imitate the cell microenvironment in bone tissue. K7M2-wt
osteosarcoma cells were seeded onto the scaffolds (15x106
cells/cm3
of scaffold volume) and
cultivated under static conditions for 1 day to allow firm cell adherence within the scaffolds. Cellseeded scaffolds were then placed in perfusion bioreactors (3D Perfuse, Innovation Center of the
Faculty of Technology and Metallurgy, Belgrade, Serbia) and cultivated for 7 days (medium flow rate
0.27 cm3/min, medium superficial velocity 40 μm/s) whereas cell cultivation under static conditions
served as a control. The cells exhibited higher metabolic activity expressing pluripotency-associated
genes when cultivated under perfusion conditions which can be attributed to the synergistic effect of
improved mass transport of nutrients and the presence of biomechanical stimuli (shear stresses up to
20 mPa). Histological analyses revealed spontaneous cell assembly into spheroid-like structures under
both static and perfusion conditions, being more prominent in the latter case. In addition, higher
quantities of extracellular matrix proteins and cytoplasmatic protein tubulin were detected in
perfused cultures. Overall, osteosarcoma cells cultivated in our 3D in vitro model demonstrate in vivolike behaviour which renders this model highly promising as a valuable tool for osteosarcoma drug
screening and tumour research.Conference Programme Book:Tumour Microenvironment Meeting & 3D Model Workshop, 6-8 September 202
PB1776: EXPRESSION OF BCL2, BAX AND MDR1 GENES AS PHARMACOTRANSCRIPTOMIC MARKERS OF PROGNOSIS IN DE NOVO PATIENTS OF ADULT ACUTE MYELOID LEUKEMIA WITH NORMAL KARYOTYPE
No full textBackground:
Acute myeloid leukemia (AML) is a malignancy of hematopoetic tissue which occurs due to a halt in
differentiation, loss of proliferation control and dysregulated apoptosis of myeloid progenitor cells. It is the most
common acute leukemia in adults, accounting for 80% of all cases. Stratification of AML patients is based on their
karyotype. However, the constant introduction of new molecular markers of prognosis of AML patients calls for a
different approach. In many cancers, as well as AML, dysregulation of apoptosis constitutes the basis of
pathogenesis and this phenomenon is important to consider when the success of chemotherapy is in question.
Another important factor of chemotherapy success is the degree of drug efflux. These factors constitute tumor
resistance. Pharmacotranscriptomic markers of tumor resistance could be important targets for specific therapy,
which, when applied, could improve prognosis in AML. The anti-apoptotic gene BCL2 (B-cell lymphoma protein 2)
and the pro-apoptotic BAX (BCL2-associated X) expression levels are markers of non-pump tumor resistance
(genes associated with apoptosis), while a marker of tumor pump resistance (gene which codes a protein associated
with drug efflux) is the expression level of MDR1 (multi-drug resistance gene 1). These tumor resistance markers
could have a significant impact on AML prognosis.
Aims:
By analyzing the gene expression profiles of BCL2, BAX and MDR1 and using statistics to identify association with
clinical indicators of prognosis in adult AML patients with normal karyotype (AML-NK), the significance of these
genes as molecular targets can be further elucidated.
Methods:
Bone-marrow samples at diagnosis were collected from 51 adult patients with AML-NK, who were treated with the
same 3+7 protocol, followed by three consolidation cycles. Expressions of BCL2, BAX and MDR1 were analyzed
using the real-time polymerase chain reaction method. Clinical data and expression values were statistically
evaluated for possible associations
Results:
The presence of chemoresistance was found to be associated with overexpression of BCL2 (BCL2+) (p=0.018), while
underexpression of BAX in patients has shown a greater affinity towards relapse (p=0.034). Evaluating the
expressions of BCL2 and BAX in a combined effect has shown that 87% of patients with BAX /BCL2low status were
resistant to therapy (p=0.024). BCL2+ status was associated with high expression of MDR1 (p<0.001). Likewise,
high expression of MDR1 was associated with the absence of NPM1 and FLT3-ITD mutations (p=0.048 and
p=0.010, respectively).
Summary/Conclusion:This is the first study that focused only on AML-NK patients, when it comes to analysis of BCL2, BAX and MDR1
gene expression profiles. The results of this preliminary study have shown that high BCL2 expression would likely
lead to tumor resistance from chemotherapy, making anti-BCL2 treatment a viable option in patients with this
expression profile. A study on a larger group of patients could clarify the prognostic importance of the studied
pharmacotranscriptomics markers, contributing to a creation of personalized treatment of adult AML-NK patients.Book of abstract: 28th Congress of the European Hematology Association EHA2023 Annual Congress Edition June 2023PB177