imagine (Institute of molecular genetics and genetic engineering)
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E-commerce readiness and training needs of small-scale dairy processors in Serbia: Understanding barriers and knowledge gaps
Get PDFThe objective of this study was to identify the requirements needed for selling dairy products through e-commerce, as well as current gaps and challenges that exist for small scale dairy processors (SSDPs), and need to be addressed in order to comply with those requirements. A mixed method research design was used for training needs assessment. Qualitative (in-depth interview with 7 online platform representatives (OPRs)) and quantitative approach (survey questionnaire with 58 SSDPs) were conducted. Interview transcripts were coded and codes were grouped into seven themes. Hierarchical cluster analysis was applied to 146 answers from 58 SSDPs. They were divided into 4 clusters. Mean sums of responses between clusters were compared by Mann-Whitney U test. OPRs suggested that SSDPs should be provided with tools and resources to help them achieve food safety and quality targets, as well as practical knowledge and skills. They reported that it is crucial to find a solution for the cold chain transportation, for maintaining consistent product quality. Survey results showed that SSDPs use kitchen equipment (79.3%) and kitchen cleaning products (81.0%) for dairy processing. In total, 43.1% process raw milk and only 24.1% have product label on the package. Only members of cluster 3 and 4 sell their products online (73.7% and 90.0%, respectively), mostly using their own social media platforms (57.9% and 60.0%, respectively), transporting products to end buyers by themselves in hand refrigerators (47.4% and 70.0%, respectively). By analyzing the differences among clusters of SSDPs, trainings can be tailored to the characteristics and knowledge gaps of each group
mRNA Vaccines Development: From Experimental Beginnings to the Nobel Prize
No full textRazvoj vakcina ima bogatu istoriju koja je obeležena revolucionarnim iskoracima i razvojem tehnologija
koje su značajno uticale na javno zdravlje. Ovaj rad prikazuje istorijski razvoj vakcina, sa posebnim fokusom
na nastanak i razvoj iRNK vakcina. Počevši od ranih inovacija u oblasti vakcina, rad pruža sveobuhvatan
pregled kako je iRNK tehnologija unela revoluciju ovu oblast. Razmatra se jedinstveni aspekt imunološkog
odgovora koji izazivaju iRNK vakcine, ističući njihovu sposobnost da proizvedu snažan i ciljan imunitet
protiv patogena. Takođe, rad analizira proces proizvodnje iRNK vakcina, objašnjavajući korake od dizajna
plazmida preko sinteze iRNK do enkapsulacije u lipidne nanopartikule. Diskusija uključuje analizu izazova
tokom faza razvoja i kako su ti izazovi prevaziđeni. Praćenjem evolucije različitih tehnologija vakcina i
naglašavanjem napretka koji su donele iRNK vakcine, ovaj rad naglašava njihov transformativni potencijal
u prevenciji infektivnih i tretmanu malignih bolesti.The development of vaccines has a rich history marked by groundbreaking innovations and evolving
technologies that have significantly improved public health. This paper explores the historical progression
of vaccines, with a particular focus on the emergence and development of mRNA vaccines. Starting from
early vaccine innovations, the study provides a comprehensive overview of how mRNA technology has
revolutionized the field. It delves into the unique aspects of the immune response elicited by mRNA
vaccines, highlighting their ability to produce robust and targeted immunity against pathogens.
Additionally, the paper examines the production process of mRNA vaccines, from plasmid design and the
synthesis of mRNA strands to their encapsulation in lipid nanoparticles. The discussion includes an analysis
of the challenges faced during the development phases and how these have been addressed. By tracing
the evolution of vaccine technology and emphasizing the advancements brought by mRNA vaccines, this
paper underscores their transformative potential in preventing infectious diseases and treating various
conditions. The findings suggest that mRNA vaccines represent a pivotal shift in vaccine development,
offering new avenues for future research and application in both infectious diseases and personalized
medicine
Characterisation of extracellular vesicles originating from thyroid cell lines and plasma of patients with thyroid nodules
No full textTiroidni karcinomi, iako najčešći endokrini maligniteti, predstavljaju dijagnostički izazov kada je u pitanju
diferencijacija malignih od benignih tumora. Drugi veliki problem tiroidne onkologije predstavlja detekcija
rekurence kod oko četvrtine oboljelih od diferenciranih tiroidnih karcinoma, kod kojih postoje
autoantitijela prema tiroglobulinu, markeru tirocita. Anti-tiroglobulinska antitijela interferiraju u testovima
za analizu tiroglobulina, onemogućavajući tačno mjerenje njegovih nivoa u cirkulaciji.
U svrhe poboljšanja dijagnostike veliki broj istraživanja bavi se ispitivanjem potencijala ekstracelularnih
vezikula (EV), što iziskuje karakterizaciju njihovih površinskih i unutarvezikularnih konstituenata. U ovoj
oblasti najveći broj istraživanja usmjeren je na mikroRNK i lncRNK molekule, dok se manji broj studija bavio
proteinskim markerima tiroidnih EV. Dosadašnji rezultati pokazuju značaj miR-146b, miR-222, lncRNK
SNHG9, kao i drugih RNK molekula, dok su predmet istraživanja proteina EV bili proteini toplotnog šoka,
kao i tiroidea-specifični proteini tirotropinski receptor (TSH-R) i tiroglobulin.
Zbog očekivane primjene EV u tiroidnoj onkologiji, postoji opravdana potreba za njihovom
karakterizacijom na prisustvo tiroidea-specifičnih površinskih i unutarvezikularnih markera, u cilju njihove
efikasnije izolacije i potencijalne dijagnostičke upotrebe.Thyroid carcinomas, although the most common endocrine malignancies, present a diagnostic challenge
in distinguishing malignant from benign tumors. Another significant issue in thyroid oncology is the
detection of recurrence in approximately a quarter of patients with differentiated thyroid carcinomas, who
are positive for autoantibodies against thyroglobulin, a marker of thyrocytes. Anti-thyroglobulin antibodies
interfere in thyroglobulin assays, rendering its analysis inaccurate.
To improve diagnostics, numerous studies are exploring the potential use of extracellular vesicles (EVs),
necessitating the characterization of their surface and intra-vesicular constituents. Most research in this
area has focused on microRNA and lncRNA molecules, while fewer studies have investigated protein
markers of thyroid EVs.
Current findings highlight the importance of miR-146b, miR-222, lncRNA SNHG9, and other RNA molecules,
while EVs protein studies have examined HSP and thyroid-specific proteins like thyrotropin receptor (TSHR)
and thyroglobulin.
Given the anticipated application of EVs in thyroid oncology, there is a justified need to characterize them
for thyroid-specific surface and intravesicular markers to enhance their isolation and potential diagnostic
use
PRIMARY CENTRAL NERVOUS SYSTEM LYMPHOMA: REAL-WORLD DATA OF 7-YEAR SINGLE-CENTER EXPERIENCE SHOW MAJORITY OF PATIENTS IN POOR PERFORMANS STATUS AND POSITIVE IMPACT OF TOTAL/MAXIMAL TUMOR RESECTION ON SURVIVAL
No full textUvod: Pacijenti sa primarnim limfomom centralnog nervnog sistema (PCNSL) se
često prezentuju ograničenom pokretljivošću i lošim opštim stanjem što ih čini
nepogodnim za kliničke studije, te je rezultate takvih studija teško tumačiti u
kontekstu svakodnevne kliničke prakse.
Cilj: Prezentovati kohortu neselektovanih pacijenata sa PCNSL iz svakodnevne
kliničke prakse i analizirati njihove kliničke karakteristike, lečenje i ishod.
Metode: Sproveli smo retrospektivnu studiju na jednom centru analizirajući
podatke prikupljene iz dokumentacije 26 pacijenata koji su lečeni u periodu 2017-
2023. godine. Pacijenti sa histologijom koja nije difuzni B-krupnoćelijski limfom
nisu uzimani u obzir.
Rezultati: Dve trećine (17, 65%) su bili slabog performans statusa (PS) (ECOG
3-4/Karnofsky ≤ 50%). Dijagnoza je postavljena nakon totalne ili maksimalne
resekcije tumora kod 12 pacijenata (46%). Preživljavanje bez progresije (PFS) je
bilo značajno duže kod pacijenata bez venske tromboembolije (VTE) (p = 0,047),
pacijenata kod kojih je na dijagnozi učinjena totalna ili maksimalna resekcija tumora
(p = 0,008), kod kojih je kompletirana indukciona terapija (p < 0,001), koji
su postigli odgovor na indukcionu terapiju (p < 0,001) i koji su lečeni konsolidacijom
zračnom terapijom (RT) ili autolognom transplantacijom (ASCT) (p < 0,001).
Totalna ili maksimalna resekcija tumora (p = 0,035), kompletiranje (p < 0,001) i
postizanje odgovora na indukcionu terapiju (p < 0,001), konsolidacija RT ili ASCT
(p < 0,001) pokazali su značajan pozitivan efekat na ukupno preživljavanje.
Zaključak: Značajan broj pacijenata u svakodnevnoj kliničkoj praksi se prezentuje
slabim PS. Značajno bolji rezultati lečenja su postignuti ako je inicijalno
sprovedena opsežna operacija, odnosno ako je sistemsko lečenje bilo maksimalno
(indukcija + konsolidacija).Introduction: Patients with primary central nervous system lymphoma (PCNSL)
often present with reduced mobility and poor general condition which make
them ineligible for clinical trials, thus leading to results that cannot be extrapolate
to real-life.
Objective: To present real-world cohort of unselected PCNSL patients and their
clinical and laboratory features, treatment and outcome.
Methods: We conducted single-center retrospective study analysing real-world
data from 26 patients managed from 2017 to 2023. Patients with histology other
than diffuse large B-cell lymphoma were excluded from the study.
Results: Two thirds (17 patients, 65%) presented with poor performance status
(PS) (ECOG 3-4/ Karnofsky index ≤ 50%). Diagnosis was made after total tumor
extirpation or maximal resection in 12 patients (46%). Progression-free survival
was significantly prolonged in patients without VTE (p = 0.047), patients who
experienced diagnostic total or maximal tumor resection (p = 0.008), in those
who completed induction treatment (p < 0.001), who achieved response after
induction treatment (p < 0.001) and received consolidative radiotherapy (RT) or
autologous stem cell transplant (ASCT) (p < 0.001). Total or maximal tumor resection
(p = 0.035), completion (p < 0.001) and response to induction treatment
(p < 0,001), consolidative RT or ASCT (p < 0.001) showed significant positive impact
on overall survival.
Conclusion: Significant number of patients in real-world setting are in poor
PS. In our cohort survival was significantly affected by VTE, total/maximal tumor
resection, completion and achievement of CR/PR to induction treatment,
and consolidation. Larger real-world databases are warranted for more reliable
conclusions.Book of abstract: NACIONALNI KONGRES 6. KONGRES HEMATOLOGA SRBIJE, NATIONAL CONGRESS 6TH CONGRESS OF HEMATOLOGISTS OF SERBIA, Hotel Crowne Plaza, Beograd
17–19. oktobar 2024. godin
Postbiotic lactobacilli induce cutaneous antimicrobial response and restore the barrier to inhibit the intracellular invasion of Staphylococcus aureus in vitro and ex vivo
No full textIntracellular pathogens including Staphylococcus aureus contribute to the non-healing phenotype of chronic wounds. Lactobacilli, well known as beneficial bacteria, are also reported to modulate the immune system, yet their role in cutaneous immunity remains largely unknown. We explored the therapeutic potential of bacteria-free postbiotics, bioactive lysates of lactobacilli, to reduce intracellular S. aureus colonization and promote healing. Fourteen postbiotics derived from various lactobacilli species were screened, and Latilactobacillus curvatus BGMK2-41 was selected for further analysis based on the most efficient ability to reduce intracellular infection by S. aureus diabetic foot ulcer clinical isolate and S. aureus USA300. Treatment of both infected keratinocytes in vitro and infected human skin ex vivo with BGMK2-41 postbiotic cleared S. aureus. Keratinocytes treated in vitro with BGMK2-41 upregulated expression of antimicrobial response genes, of which DEFB4, ANG, and RNASE7 were also found upregulated in treated ex vivo human skin together with CAMP exclusively upregulated ex vivo. Furthermore, BGMK2-41 postbiotic treatment has a multifaceted impact on the wound healing process. Treatment of keratinocytes stimulated cell migration and the expression of tight junction proteins, while in ex vivo human skin BGMK2-41 increased expression of anti-inflammatory cytokine IL-10, promoted re-epithelialization, and restored the epidermal barrier via upregulation of tight junction proteins. Together, this provides a potential therapeutic approach for persistent intracellular S. aureus infections
Pharmacogenomic landscape of Serbian population
No full textMany drugs account for an extensive interpatient variability in dose requirement for
achieving therapeutic benefit. Said variability is often largely influenced by common genetic
variants in several pharmacogenes. Herein we present an overview of pharmacogenomic
landscape of Serbian population on the example of 7 actionable pharmacogenes for
which Clinical Pharmacogenetics Implementation Consortium issued guidelines for
interpretation and use of haplotypes to estimate adequate dose in therapeutics for
treatment of cancer, rheumatic and cardiovascular diseases: TPMT, SLCO1B1, NUDT15,
UGT1A1, VKORC1, CYP2C9 and CYP4F2.
A cohort of 113 patients from Serbia underwent whole exome sequencing. Generated
FASTQ files were aligned to human reference genome build GRCh38 and variants were
annotated using in-house pipeline. Calling of star alleles was performed using opensourced
bioinformatics tool Stargazer applied to BAM and VCF files. Subsequent analysis
of frequency differences in Serbian versus European population was done using 1000
Genomes Project database and Chi square test in R program.
In 7 analyzed pharmacogenes 23 different haplotypes, denoted as star alleles, were
found in total. Nine of these haplotypes had significantly different frequencies in Serbian
comparing to European population – CYP4F2*2, variant rs9923231 T in VKORC1, TPMT*20
and TPMT*43, UGT1A1*36 and UGT1A1*80, while in actionable pharmacogene SLCO1B1
three haplotypes (*4, *20 and *37) with significantly higher frequencies in Serbian than in
European population were detected.
Presented results show that frequencies of several important pharmacogenomics star
alleles differ in Serbian comparing to European population. This could have serious clinical
implications and potentially lead to more frequent development of adverse drug reactions
when treating patients in need with standard drug doses. In Serbian population a potential
for preemptive genotype testing exist when prescribing therapeutics for cancer (TPMT,
NUDT15, UGT1A1, SLCO1B1) and rheumatic (SLCO1B1, TPMT) or cardiovascular diseases
(VKORC1, CYP2C9, CYP4F2), and population-specific pharmacogenomics aspect should be
taken into account for therapy optimization in clinical practiceBook of abstracts: 5th Belgrade Bioinformatics Conference, Serbia, Belgrade,17-20 june 2024
Non-coding transcripts of protein-coding genes as novel biomarkers for colorectal cancer diagnosis
No full textRecent research shows that non-coding RNA transcripts of protein-coding genes could
be an emerging novel class of diagnostic biomarkers. This study aimed to identify the
most promising biomarkers for colorectal cancer (CRC) screening among upregulated
non-coding transcripts of protein-coding genes in malignant CRC cells in comparison to
non-malignant cells grown in 3D. Malignant CRC cell lines HCT116, DLD-1 and SW620,
and a non-malignant human colon epithelial cell line HCEC-1CT, were cultivated in 3D
as spheroids in 24-well plates with low attachment surface for 7 days. RNA sequencing
was performed on ribosomal-depleted total RNA using Illumina’s NovaSeq6000 platform
that generated paired-end 150bp reads. Highly upregulated transcripts (>10 FPKM)
present in all malignant cell lines and absent in non-malignant cell line were filtered
and analyzed by a set of in silico tools to filter the best candidates for further validation
studies. The publicly available GSE164541 set consisting of triplicate tissue samples of
normal, adenoma and primary CRC tissues collected from five patients with CRC was
used for validation. As a result, 5 transcripts with retained introns ANXA3-204, LLGL2-
207, KRT19-204, KRT18-206 and KRT8-213 were analyzed by in silico tools. Only
ANXA3-204 was classified as non-coding according to both CPC2 and LGC online coding
potential prediction tools. Nucleus was predicted as subcellular localization for ANXA3-
204 by AnnoLnc2. Additionally, ANXA3-204 expression was upregulated in adenoma
(p=0.04) and CRC tissue samples, although statistical significance was not reached for
CRC, in comparison to normal tissue samples in the validation set. Transcriptomic analysis
revealed that non-coding ANXA3-204 transcript was highly upregulated in malignant CRC
cell lines and adenomas compared to control samples, making ANXA3-204 a potential
candidate for early CRC screening. Further studies are needed to confirm diagnostic
potential and regulatory role of ANXA3-204.Book of abstracts: 5th Belgrade Bioinformatics Conference, Serbia, Belgrade,17-20 june 2024
ENHANCING GUT HEALTH AND LONGEVITY THROUGH NOVEL STARTER CULTURES IN FERMENTED PRODUCTS
No full textFor nearly 10,000 years, fermented foods have been essential to the human diet, boasting a
significant variety today. The health advantages of fermented foods have been recognized for a long
time. These benefits may result from direct interactions of ingested live microorganisms with the
host, as a probiotic effect. They may also come indirectly as a result of the ingestion of microbial
metabolites synthesized during fermentation. Research suggests that probiotics found in dairy
products have positive effects on human health. In recent years, there has been extensive research
into the use of starter cultures with probiotic characteristics to address various health conditions. A
Western-type diet characterized by a high daily intake of saturated fats and refined carbohydrates
leads to the accumulation of a wide variety of molecular and cellular damages over time, leading to
several age-related diseases. According to the World Health Organization, the number of people
worldwide over 60 years of age was estimated to be 1 billion in 2019, with an expected increase to
2.1 billion by 2050. The increasing number of the elderly population will be accelerated in the
coming decades, particularly in developing countries, such as Serbia. We have tested several
carefully selected natural isolates of lactic acid bacteria, originating from artisanal dairy products
from specific geographical locations in the Balkan peninsula for the ability to decelerate the cellaging
process. Our results revealed that these strains possess exciting probiotic features such as
strengthening the epithelial intestinal barrier through stimulation of autophagy, upregulating the
tight junctions between the epithelial cells of the intestine which prevent the passage of harmful
substances from the intestine to other organs, activating the antimicrobial defense, and extending
the lifespan of Caenorhabditis elegans via autophagy activation, making them great candidates for
probiotic starter cultures for functional dairy food.Book of Abstracts : The 3rd International UNIFood Conference, UNIFood2024 Conference, Belgrade, June 28-29, 202
Cutting-Edge iPSC-Based Approaches in Studying Host—Microbe Interactions in Neuropsychiatric Disorders
No full textGut microbiota (GM), together with its metabolites (such as SCFA, tryptophan, dopamine, GABA, etc.), plays an important role in the functioning of the central nervous system. Various neurological and psychiatric disorders are associated with changes in the composition of GM and their metabolites, which puts them in the foreground as a potential adjuvant therapy. However, the molecular mechanisms behind this relationship are not clear enough. Therefore, before considering beneficial microbes and/or their metabolites as potential therapeutics for brain disorders, the mechanisms underlying microbiota–host interactions must be identified and characterized in detail. In this review, we summarize the current knowledge of GM alterations observed in prevalent neurological and psychiatric disorders, multiple sclerosis, major depressive disorder, Alzheimer’s disease, and autism spectrum disorders, together with experimental evidence of their potential to improve patients’ quality of life. We further discuss the main obstacles in the study of GM–host interactions and describe the state-of-the-art solution and trends in this field, namely “culturomics” which enables the culture and identification of novel bacteria that inhabit the human gut, and models of the gut and blood–brain barrier as well as the gut–brain axis based on induced pluripotent stem cells (iPSCs) and iPSC derivatives, thus pursuing a personalized medicine agenda for neuropsychiatric disorders
Effect of Type-2 Diabetes Mellitus on the Expression and Function of Smooth Muscle ATP-Sensitive Potassium Channels in Human Internal Mammary Artery Grafts
No full textHere we have shown for the first time altered expression of the vascular smooth muscle (VSM) KATP channel subunits in segments of the human internal mammary artery (HIMA) in patients with type-2 diabetes mellitus (T2DM). Functional properties of vascular KATP channels in the presence of T2DM, and the interaction between its subunits and endogenous ligands known to relax this vessel, were tested using the potassium (K) channels opener, pinacidil. HIMA is the most commonly used vascular graft in cardiac surgery. Previously it was shown that pinacidil relaxes HIMA segments through interaction with KATP (SUR2B/Kir6.1) vascular channels, but it is unknown whether pinacidil sensitivity is changed in the presence of T2DM, considering diabetes-induced vascular complications commonly seen in patients undergoing coronary artery bypass graft surgery (CABG). KATP subunits were detected in HIMA segments using Western blot and immunohistochemistry analyses. An organ bath system was used to interrogate endothelium-independent vasorelaxation caused by pinacidil. In pharmacological experiments, pinacidil was able to relax HIMA from patients with T2DM, with sensitivity comparable to our previous results. All three KATP subunits (SUR2B, Kir6.1 and Kir6.2) were observed in HIMA from patients with and without T2DM. There were no differences in the expression of the SUR2B subunit. The expression of the Kir6.1 subunit was lower in HIMA from T2DM patients. In the same group, the expression of the Kir6.2 subunit was higher. Therefore, KATP channels might not be the only method of pinacidil-induced dilatation of T2DM HIMA. T2DM may decrease the level of Kir6.1, a dominant subunit in VSM of HIMA, altering the interaction between pinacidil and those channels