imagine (Institute of molecular genetics and genetic engineering)
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Neurorazvojni poremećaji kod dece sa 22q11.2 delecijskim sindromom i preporuke za pedijatrijsko praćenje
No full textNeurodevelopmental disorders are the most prevalent chronic diagnoses in pediatric primary care, with rising incidence and significant impact on cognitive, motor, social, and communication functioning. 22q11.2 deletion syndrome (22q11.2 DS)-the most common human microdeletion syndrome-presents with a broad spectrum of somatic and neurodevelopmental abnormalities. Nearly all individuals with 22q11.2 DS show neurodevelopmental difficulties, including delays in motor and speech milestones, cognitive impairments, and behavioral challenges. The disorder affects approximately 1 in 2,500 newborns and is also associated with congenital heart defects, palatal anomalies, hypocalcemia, and immunodeficiency. Neurodevelopmental manifestations typically begin in infancy with delayed motor and speech development and progress into school age with difficulties in learning, attention, and peer interaction. Intellectual disabilities are common, with a distribution skewed toward lower IQ scores. Children often exhibit a verbal-performance IQ discrepancy and may experience further cognitive decline in adolescence or adulthood. Over 40% of affected individuals meet criteria for autism spectrum disorder, attention-deficit/hyperactivity disorder, or both. They also have increased risks for psychiatric conditions such as anxiety, depression, and schizophrenia. Pediatricians, as primary care providers, play a critical role in early identification and long-term monitoring. Recommendations include routine developmental assessments, early interventions (e.g., speech and occupational therapy), and regular IQ and adaptive functioning evaluations, especially during educational transitions. Early diagnosis and individualized, multidisciplinary approaches are essential to improve developmental outcomes and quality of life in children with 22q11.2 DS.Neurorazvojni poremećaji predstavljaju najčešće hronične dijagnoze u primarnoj pedijatrijskoj zaštiti, sa sve većom učestalošću i značajnim uticajem na kognitivno, motoričko i socijalno funkcionisanje. Sindrom delecije 22q11.2 (22q11.2 DS) — najčešći mikrodelecijski sindrom kod ljudi — karakteriše širok spektar somatskih i neurorazvojnih abnormalnosti. Gotovo svi pojedinci sa 22q11.2 DS pokazuju neurorazvojne poteškoće, uključujući kašnjenje u razvoju motorike i govora, kognitivna oštećenja i probleme u komunikaciji i ponašanju. Ovaj poremećaj pogađa otprilike jedno od 2.500 novorođenčadi i povezan je i sa urođenim srčanim manama, anomalijama nepca, hipokalcemijom i imunodeficijencijom. Neurorazvojne manifestacije najčešće počinju u ranom detinjstvu kašnjenjem u razvoju motorike i govora, a nastavljaju se u školskom uzrastu kroz poteškoće u učenju, pažnji i socijalnim interakcijama. Intelektualne teškoće su česte, sa distribucijom učestalosti pomerenom ka nižim vrednostima koeficijenta inteligencije. Deca često pokazuju razliku između verbalnog i neverbalnog skora i mogu imati dodatni kognitivni pad tokom adolescencije ili odraslog doba. Više od 40% pacijenata ispunjava kriterijume za poremećaj iz spektra autizma, poremećaj pažnje/hiperaktivnosti, ili oba. Takođe imaju povećan rizik od psihijatrijskih stanja kao što su anksioznost, depresija i šizofrenija. Pedijatri, kao lekari primarne zdravstvene zaštite, imaju ključnu ulogu u ranom prepoznavanju i dugoročnom praćenju. Preporuke za praćenje uključuju rutinske razvojne procene, rane razvojne intervencije (npr. govornu i radnu terapiju) i redovno procenjivanje koeficijenta inteligencije i adaptivnog funkcionisanja, naročito tokom prelaznih obrazovnih perioda. Rana dijagnoza i individualizovani, multidisciplinarni pristupi su od suštinskog značaja za unapređenje razvojnih ishoda i kvaliteta života dece i odraslih sa 22q11.2 DS
DEVELOPMENT OF IPSC-BASED MODEL SYSTEM FROM PATIENTS WITH 22Q11.2 DUPLICATION SYNDROME FOR STUDYING NEURODEVELOPMENTAL DISORDERS
No full text22q11.2 Duplication Syndrome (22q11.2DupS) is associated with an elevated risk of developing
neurodevelopmental disorders (NDDs), such as autism spectrum disorders (ASD) and
attention-deficit/hyperactivity disorder. ASD is detected in 14-25% of patients, making
22q11.2DupS one of the genetic syndromes with the highest prevalence of ASD. To better
understand the molecular mechanisms underlying the pathophysiology of NDDs, we have
generated induced pluripotent stem cells (iPSCs) from patients with 22q11.2DupS. Mononuclear
cells from 22q11.2DupS patients and healthy individuals were reprogrammed into iPSC lines
using the CytoTuneTM-iPS 2.0 Sendai Reprogramming Kit. The established iPSC lines were
characterized by examining the expression of stem cell markers through RT-PCR and their
ability to differentiate into cells of three germ layers using the STEMdiffTM Trilineage
Differentiation Kit. We differentiated the iPSCs into neurons using the dual SMAD inhibition
method together with retinoic acid. The expression of neural and neuronal-specific markers in
neural progenitors and mature neurons was analyzed using RT-PCR and immunocytochemistry.
The established iPSC lines from three patients with an inherited form of 22q11.2DupS, their
mothers who carry the microduplication, and three healthy individuals demonstrated the
expression of stem cell markers and the ability to differentiate into the cells of all three germ
layers. Both iPSCs from carriers of the 22q11.2 microduplication and healthy controls
successfully differentiated into neurons, which was confirmed by the expression of neuronal
markers. The generated 22q11.2DupS iPSC lines provide a valuable resource for understanding
the molecular mechanisms underlying NDDs associated with 22q11.2 microduplication.Abstract book: FENS Regional Meeting 2025 , Oslo, Norway, 16-19 June 202
Effects of Decreased SOX3 Gene Expression on the Properties of U251 Glioblastoma Cells
No full textGliomi su primarni tumori centralnog nervnog sistema i smatra se da nastaju od
neuralnih matičnih ili progenitornih ćelija koje imaju tumor-inicirajuće genetičke promene
(1). Oni su među najčešćim tumorima mozga i čine oko 30% svih primarnih i 80% svih
malignih tumora mozga. Gliomi su heterogeni tumori i uzrok većine smrtnih ishoda
izazvanih primarnim tumorima mozga (1). Glavne osobine glioma su visoka stopa
maligniteta i smrtnosti, kao i povećan rizik od ponovnog javljanja (2).
Incidenca glioma u Norveškoj iznosi 7,4 slučaja na 100.000 ljudi godišnje (3).
Incidenca je viša kod muškaraca (8,8/100.000) nego kod žena (6,1/100.000), što odgovara
odnosu polova od 1,4:1 (3). Prisustvo varijacija po regionima je takođe zapaženo - stopa
incidence glioma u Japanu je više nego dvostruko niža u poređenju sa Severnom Evropom
i Sjedinjenim Američkim Državama (4-6), iako uzroci ovih razlika nisu poznati. Incidenca
glioma značajno raste sa starošću, naročito kada je reč o glioblastomu. Na 100.000 ljudi,
incidenca glioblastoma iznosi 0,15 kod dece, 0,41 kod mladih, 13,1 kod osoba uzrasta 65–
74 godina, te 15 kod osoba između 75 i 84 godine (https://oncohemakey.com/intracranial-
tumors/). Iako direktni uzroci porasta rizika sa godinama još uvek nisu u potpunosti
razjašnjeni, starenje se smatra opštim faktorom rizika za razvoj mnogih malignih bolesti
Analiza kvaliteta sekvenciranja celog genoma korišćenjem humane DNK izolovane iz krvi i pljuvačke
No full textСеквенцирање ДНК је процес којим се одређује редослед нуклеотида у ДНК молекулу.
Прва метода за секвенцирање ДНК, метода секвенцирања ДНК по Сангеру, објављена је
1977. године и од тада је у широкој примени (1). Ова метода секвенцирања представља
прву генерацију секвенцирања и заснива се на ензимској синтези ДНК, коришћењу
обележених (радиоактивно или флуоресцентно) дидеокси нуклеотида који представљају
терминаторе синтезе и електрофоретском раздвајање фрагмента ДНК.
У периоду између 2004. и 2006. године дошло је до развоја метода нове генерације
секвенцирања које се заснивају на паралелној анализи великог броја сегмената ДНК
(масовно паралелно секвенцирање) и потом повезивању добијених података
биоинформатичком анализом података (2).
Методе друге генерације секвенцирања заснивају се на масовном паралеленом
секвенцирању милиона кратких секвенци величине од 250 до 800 базних парова (2).
Процес обухвата припрему библиотеке, секвенцирање и обраду добијених података.
Недостатак методе друге генерације секвенцирања је немогућност детектовања више од
70% хуманих структурних геномских варијанти (2).
Методе треће генерације секвенцирања заснивају на масовном паралеленом
секвенцирању дугих секвенци (енг. long-read sequencing) дужине од једне килобазе до
неколико мегабаза (2).
У зависности да ли је материјал који ће бити секвенциран ДНК или РНК, секвенцирање
нове генерације може се најшире поделити на секвенцирање ДНК (енг. DNA-seq) и
секвенцирање РНК (енг. RNA-seq). Секвенцирање ДНК се надаље дели на секвенцирање
целог генома (енг. Whole Genome Sequencing - WGS), секвенцирање целог егзома (енг.
Whole Exome Sequencing - WES), секвенцирање епигенома (енг. Epigenome Sequencing) и
циљано (таргетно) секвенцирање (енг. Targeted Sequencing) (2).Sekvenciranje DNK je proces kojim se određuje redosled nukleotida u DNK molekulu.
Prva metoda za sekvenciranje DNK, metoda sekvenciranja DNK po Sangeru, objavljena je
1977. godine i od tada je u širokoj primeni (1). Ova metoda sekvenciranja predstavlja
prvu generaciju sekvenciranja i zasniva se na enzimskoj sintezi DNK, korišćenju
obeleženih (radioaktivno ili fluorescentno) dideoksi nukleotida koji predstavljaju
terminatore sinteze i elektroforetskom razdvajanje fragmenta DNK.
U periodu između 2004. i 2006. godine došlo je do razvoja metoda nove generacije
sekvenciranja koje se zasnivaju na paralelnoj analizi velikog broja segmenata DNK
(masovno paralelno sekvenciranje) i potom povezivanju dobijenih podataka
bioinformatičkom analizom podataka (2).
Metode druge generacije sekvenciranja zasnivaju se na masovnom paralelenom
sekvenciranju miliona kratkih sekvenci veličine od 250 do 800 baznih parova (2).
Proces obuhvata pripremu biblioteke, sekvenciranje i obradu dobijenih podataka.
Nedostatak metode druge generacije sekvenciranja je nemogućnost detektovanja više od
70% humanih strukturnih genomskih varijanti (2).
Metode treće generacije sekvenciranja zasnivaju na masovnom paralelenom
sekvenciranju dugih sekvenci (eng. long-read sequencing) dužine od jedne kilobaze do
nekoliko megabaza (2).
U zavisnosti da li je materijal koji će biti sekvenciran DNK ili RNK, sekvenciranje
nove generacije može se najšire podeliti na sekvenciranje DNK (eng. DNA-seq) i
sekvenciranje RNK (eng. RNA-seq). Sekvenciranje DNK se nadalje deli na sekvenciranje
celog genoma (eng. Whole Genome Sequencing - WGS), sekvenciranje celog egzoma (eng.
Whole Exome Sequencing - WES), sekvenciranje epigenoma (eng. Epigenome Sequencing) i
ciljano (targetno) sekvenciranje (eng. Targeted Sequencing) (2)
DOGS AS RESERVOIRS OF ECHINOCOCCUS SPP. TAPEWORMS AND RELEVANCE FOR TRANSMISSION TO HUMANS
No full textObjectives: Dogs are definitive hosts for Echinococcus spp. tapeworms. To assess the
relevance of hunting and stray dogs as reservoirs of Echinococcus spp. eggs, which are
infective for humans, feces samples were collected from several locations in Serbia with
a confirmed presence of Echinococcus spp. in other animals. None of the dogs received
anticestodal treatment prior to feces collection.
Materials and methods: Taeniid eggs were isolated using a combined flotation and
mesh filtration technique. Total gDNA was extracted from the collected eggs and
multiplex nested PCR, based on detection of specific Cox1 sequences of E. multilocularis,
E. granulosus and E. canadensis, was performed.
Results: Thus far, n = 30 samples of hunting dog and n = 120 samples of stray dog feces
were collected. Echinococcus spp. eggs were not present in any of the hunting dog
samples. Further molecular screening for other cyclophillidean tapeworms revealed two
positive dogs.
Conclusion: Despite the absence of regular anticestodal treatment and distinct
possibility of exposure to Echinococcus spp., none of the hunting dogs were shedding
tapeworm eggs. These preliminary findings suggst that hunting dogs may not be relevant
reservoirs of Echinococcus spp. for human infection. Processing of additional samples is
underway.Book of abstract: 57th Days of Preventive Medicine, International Congress Contemporary Challenges in Public Health, 23-26. September 2025. Niš, Serbi
67 Indukovane pluripotentne matične ćelije kao model za proučavanje pankreatitisa izazvanog lekovima: prošlost, sadašnjost i budućnost
No full textDrug-induced pancreatitis (DIP) represents a significant clinical challenge, accounting for approximately
0.1-2% of all acute pancreatitis cases. Among immunosuppressive therapies, thiopurines are particularly
notorious for causing pancreatitis, with an incidence of 3-5% in inflammatory bowel disease (IBD) patients.
The idiosyncratic and dose-independent nature of thiopurine-induced pancreatitis (TIP) has hindered the
development of predictive biomarkers and mechanistic understanding. Traditional in vitro models using
immortalized cell lines and animal models have proven insufficient to recapitulate the complexity of
human pancreatic responses to drugs. The advent of induced pluripotent stem cell (iPSC) technology has
opened new avenues for personalized disease modeling and drug toxicity assessment. This review explores
the evolution of iPSC-based models for studying drug-induced pancreatitis, from early bidimensional
cultures to sophisticated three-dimensional organoids, discussing their applications in elucidating
pharmacokinetic and pharmacodynamic mechanisms, identifying genetic risk factors, and developing
precision medicine approaches. Recent breakthrough studies demonstrate that iPSC-derived pancreatic
models from patients who developed TIP show enhanced sensitivity to thiopurine cytotoxicity, with distinct
mechanisms involving TPMT expression in stem cells and Rac1 protein levels in differentiated pancreatic
cells. These findings highlight the potential of iPSC technology for predictive toxicology and therapy
personalization, particularly in vulnerable pediatric populations where clinical trials are limited by ethical
constraints
Analiza podataka sekvence celog genoma ćelijskog klona U937 sa uklonjenim genom ZBTB14 i funkcije gena ZBTB14
No full textHumani gen ZBTB14 je smešten na hromozomu 18, na poziciji 18p11.21(GRCh38/hg38) i
obuhvata 8035 baznih parova (bp). Takođe, gen ZBTB14 je poznat pod nazivom Zinc finger
protein 161 (ZFP161) i Zinc finger 5 (ZF5). Ovaj gen se sastoji od dva egzona, koja su
razdvojena intronom, pri čemu se prva tri kodirajuća nukleotida nalaze u prvom egzonu, a
ostatak kodirajuće sekvence se nalazi u drugom egzonu. Gen ZBTB14 obuhvata otvoreni
okvir čitanja od 1347 bp i kodira protein ZBTB14.
Geni koji kodiraju proteine sa cinkanim prstima, kao što su Only Zinc Finger (OZF), Zinc
finger protein 134 (ZNF134) i ZNF135, smešteni su u klasteru na hromozomu 19, i imaju
visok stepen homologije sa sekvencom gena ZBTB14. Stoga se pretpostavlja da je gen
ZBTB14 evoluirao iz zajedničkog predačkog gena, to jest da se tokom evolucije odvojio od
drugih proteina sa cinkanim prstima (1)
Genomic profiling, implications for genotype-based treatment of 131 patients with phenylketonuria and characterization of novel p.Pro416Leu PAH variant
No full textPhenylketonuria (PKU) is the most common inborn disorder of amino acid metabolism caused by biallelic pathogenic variants in phenylalanine hydroxylase (PAH) gene. This study comprised genomic profiling and phenotypic characterization of 131 Serbian PKU patients along with implications for BH4 therapy. By combining Sanger sequencing, MLPA and WES re-analysis of previously unsolved cases, we identified 38 different disease causing variants in the PAH gene and classified them using ACMG guidelines. The most frequent variant was p.Leu48Ser (30.92%), followed by p.Arg408Trp (12.21%) and p.Ile306Val (8%). We detected one novel variant, p.Pro416Leu, which was classified as pathogenic, based on computational algorithms prediction, with destabilization as the mechanism of the effect upon PAH protein. For patients’ phenotypic classification, we used pre-treatment serum Phe level and Phe tolerance, indicating that 42% of patients had classic PKU, 22% had mild PKU and 32% were classified as MHP (4% remained unclassified). Furthermore, we performed genotypephenotype correlation study which emphasized the inconsistency of p.Leu48Ser variant. Given that not all PKU patients benefit from genotype-based therapy (Kuvan and sepiapterin), we assessed the potential responsiveness in our patients. We categorized all detected PAH genotypes accordingly and with 39.1% responsive and 44.5% probably responsive found that majority of patients may respond to BH4 therapy. Our study brings the updated spectrum of molecular genetic data, variant classification, characterization of novel p.Pro416Leu variant, detailed phenotypic characteristics and BH4 responsiveness for PKU patients from Serbia, therefore contributing to better understanding of molecular landscape of PKU.Supplementary Information:[https://doi.org/1 0.1038/s41598-025-04611-2
Bacterial exopolysaccharides mediate activation of flavin-containing monooxygenase 2 to extend lifespan in Caenorhabditis elegans
No full textThe gut microbiota plays a pivotal role in modulating host physiology and longevity through the production of microbial-derived molecules. Among these, bacterial exopolysaccharides (EPS) represent a structurally diverse group of surface polysaccharides with emerging roles in regulating host well-being. Here, we investigated the role of Lactobacillus strains with the capability to produce EPS, using Caenorhabditis elegans as a model organism. Results revealed significant lifespan extension in worms fed with EPS-producing bacteria, accompanied by improved health-span markers such as enhanced pharyngeal pumping and reduced lipofuscin accumulation. Transcriptomic profiling identified robust upregulation of the host detoxification and immune defense pathways, highlighting the flavin-containing monooxygenase gene fmo-2, as one of the major mediators of longevity and stress resistance triggered by EPS-producing lactobacilli. The effect was confirmed using fmo-2p::GFP reporter animals and was abrogated in fmo-2, hlh-30, and nhr-49 mutant backgrounds. Mechanistically, we demonstrated that EPS acts through a conserved transcriptional network that primarily relies on the activation of nhr-49/PPAR-α, with purified EPS being sufficient to activate fmo-2 expression. Our findings reveal that bacterial EPS activates host xenobiotic pathways to modulate aging, positioning it as a potential tool for microbiota-based longevity interventions. These insights show how microbial products can modulate fundamental biological processes across species, opening new strategies for age-related health interventions
Arabidopsis Cold Stress Tolerance Improvement via AthHOS1-targeting HOS1-amiRNA Approach
No full textIn this study, we examined the efficacy of the artificial microRNAs (amiRNAs)
technology in targeting the HOS1 gene for the enhancement of cold stress tolerance in
Arabidopsis thaliana Ler-0 ecotype. The impact of athHOS1-amiRNA overexpression on
the response of transgenic plants to cold stress was assessed using RT-qPCR in 3-week-old
seedlings of the T3 generation. Additionally, the response of wild-type plants of the same
age to cold stress (4oC) for various durations (6, 12, 24, 48, and 96 hours) was also
evaluated. Comparative analysis revealed that athHOS1-amiRNA downregulated
athHOS1 in transgenic plants after prolonged exposure to low temperature (48 h and 96
h) (Pearson’s correlation coefficient of -0.407; P< 0.05). Interestingly, while prolonged
cold stress at 96 h led to a significant upregulation of athHOS1 in wild-type plants, the
suppression of athHOS1-amiRNA in transgenic plants disrupted the expected circadian
rhythm of athHOS1 by preventing its upregulation. Furthermore, T3 plants that had been
cold-acclimated exhibited a 17% increase in freezing tolerance (-1 to -8°C) compared to
wild-type plants, indicating the success of this approach in enhancing Arabidopsis
tolerance to low temperatures, at least in the Ler-0 ecotype. In order to gain a deeper
understanding of the intricate dynamics of gene/protein networking during cold
acclimation and its interaction with the athHOS1-amiRNA approach, further
characterization is required. This includes measuring the expression levels and half-life of
athHOS1-amiRNA and HOS1 mRNA, as well as evaluating the protein level of HOS1 and
its direct targets, such as ICE1, in different Arabidopsis ecotypes and at different time
intervals of low temperature exposure