Background: Array‐based genomic analysis is a gold standard for the detection of copy number variations (CNVs) as an important source of benign as well as pathogenic variations in humans. The introduction of chromosomal microarray (CMA) has led to a significant leap in diagnostics of genetically caused congenital malformations and neurodevelopmental disorders, with an average diagnostic yield of 15%. Here, we present our experience from a single laboratory perspective in four years’ postnatal clinical CMA application.Methods: DNA samples of 430 patients with congenital anomalies and/or neurodevelopmental disorders were analyzed by comparative genome hybridization using oligonucleotide‐based microarray platforms. Interpretation of detected CNVs was performed according to current guidelines. The detection rate (DR) of clinically significant findings (pathogenic/likely pathogenic CNVs) was calculated for the whole cohort and isolated or combined phenotypic categories.Results: A total of 140 non‐benign CNVs were detected in 113/430 patients (26.5%). In 70 patients at least one CNV was considered clinically significant thus reaching a diagnostic yield of 16.3%. The more complex the phenotype, including developmental delay/intellectual disability (DD/ID) as a prevailing feature, the higher the DR of clinically significant CNVs is obtained. Isolated congenital anomalies had the lowest, while the “dysmorphism plus” category had the highest diagnostic yield.Conclusion: In our study, CMA proved to be a very useful method in the diagnosis of genetically caused congenital anomalies and neurodevelopmental disorders. DD/ID and dysmorphism stand out as important phenotypic features that significantly increase the diagnostic yield of the analysis.

Objectives: Inflammatory cytokines like tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta, and IL-6 can cause brain injury, slow recovery, and adverse effects (ADEs) in ischemic stroke (IS) patients treated with recombinant tissue plasminogen activator (rtPA). We explored the relationship between selected polymorphisms within TNF-alpha, IL-1 beta and IL-6 genes, and post-IS outcome and ADEs in patients treated with rtPA.Methods: One hundred and sixty-six patients with IS treated with rtPA were included in this study. The modified Rankin Scale (mRS) was used to assess functional recovery 3 months after IS likewise thrombolytic therapy efficacy. Patients were classified into groups with favorable (0-1) or poor recovery based on their mRS score at the ninetieth day post-IS. During hospitalization, ADEs following rtPA were monitored. TNF-alpha-308 G/A (rs1800629), IL-1 beta-511 G/A (rs16944), and IL-6-174 G/C (rs1800795) polymorphisms were genotyped using Real-Time PCR. SPSS software version 22.0 was used for statistical analyses.Results: Patients with the TNF-alpha-308 G/A GG genotype had a higher mean NIHSS value at admission (12.75 +/- 5.176) than those carrying A-allele (10.56 +/- 3.979;p = 0.016). Individuals with the CC genotype of the IL-6-174 G/C polymorphism had significantly lower NIHSS scores (8.79 +/- 5.053) than those with G-allele (12.06 +/- 6.562) 24 hours after rtPA (p = 0.050). Patients with the GG genotype of the IL-6-174 G/C polymorphism had a significantly poorer outcome (p = 0.024; OR = 2.339; 95%CI 1.121-4.880), while patients who were G-allele carriers of the Il-6-174 G/C polymorphism and had the AA genotype of the IL-1 beta-511 G/A polymorphism were statistically significantly more likely to experience hemorrhagic transformation (p = 0.046; OR = 2.7273; 95%CI 1.0414-7.1426).Conclusion: GG genotype of the IL-6-174 G/C polymorphism is associated with poor recovery after IS treated with rtPA therapy.

Secondary peritonitis (SP) remains a major clinical challenge due to its high complication rates and it often results in sepsis and multi-organ dysfunction. This study investigated the association between four nitric oxide synthase (NOS) single-nucleotide polymorphisms (SNPs)—NOS3 c.-786T>C (rs2070744), NOS3 c.894G>T (rs1799983), NOS3 27 bp variable number tandem repeat (VNTR) (rs61722009), and NOS2 (rs2297518)—and sepsis-related complications in 202 patients with SP. Demographic and baseline clinical characteristics, Acute Physiology and Chronic Health Evaluation (APACHE) II scores, Mannheim Peritonitis Index, and complications (multiple organ dysfunction syndrome (MODS), multiple organ failure (MOF), acute respiratory distress syndrome (ARDS), and sepsis) were analyzed for associations with the NOS gene variants. Haplotype analysis was also performed. No SNP showed an association with in-hospital mortality. However, the NOS3 c.-786T>C TT genotype was significantly associated with an increased risk of MOF (p = 0.008), and remained independently associated after multivariate adjustment (p_MOF = 0.006). The T4bG haplotype was significantly more frequent among patients with MODS (p = 0.026), MOF (p = 0.048), and sepsis (p = 0.018). These findings suggest that NOS gene variants, particularly NOS3 c.-786T>C and the T4bG haplotype, may potentially serve as biomarkers for risk stratification in critically ill patients.

Background/Objectives: Hypoxic–ischemic encephalopathy (HIE) is a common neurological outcome of perinatal asphyxia, with cerebral palsy (CP) being the most severe lasting effect. Perinatal brain injury activates the immune system and induces the release of inflammatory mediators. Matrix Metalloproteinases (MMPs) play a crucial role in neuroinflammation and neurodegeneration. This study explored the potential link between MMP2 promoter polymorphisms and the development of CP in children with a history of perinatal asphyxia. Methods: We enrolled 212 patients (130 males and 82 females) with documented perinatal asphyxia, who underwent a comprehensive neurological assessment and neuroimaging, including ultrasound and magnetic resonance imaging (MRI). We genotyped the MMP2 promoter polymorphisms rs243866, rs243865, and rs243864 using real-time polymerase chain reaction. Haplotype frequencies were calculated using Haploview software. Results: As expected, patients with HIE are more likely to develop CP (p = 0.000). In a study of 104 patients who developed CP, the frequencies of the A (rs243866), T (rs243865), and G alleles (rs243864) were nearly twice as high compared to those without CP (p = 0.008, p = 0.019, and p = 0.008, respectively). Haplotype analysis supported these findings, showing that the ATG haplotype was significantly more common among patients who developed CP (p = 0.004). Additionally, in patients with MRI-confirmed brain damage, the ATG haplotype was more frequently observed (p = 0.019). Conclusions: The ATG haplotype of the MMP2 promoter may indicate a risk factor for developing cerebral palsy (CP) in patients who experience perinatal asphyxia and could serve as a potential diagnostic predictor of CP.

Poznato je da se neurorazvojni poremećaji (NRP) češće javljaju kod dečaka. U zavisnosti od specifične dijagnoze, odnos muškog i ženskog pola u ukupnom oboljevanju može prevazići 4:1. Jedno od mogućih objašnjenja ovog fenomena jeste da su osobe ženskog pola biološki zaštićene i moraju imati veće genetičko opterećenje da bi ispoljile NRP nego dečaci, što čini osnovu teorije protektivnog efekta ženskog pola. Neke studije su pokazale veću učestalost retkih de novo mutacija, posebno patogenih varijacija u broju kopija u genomu (CNV, copy number variation) kod devojčica sa ispoljenim NRP u odnosu na dečake sa istim poremećajem. U našem istraživanju ispitivali smo polne razlike u prevalenciji fenotipova i stopama detekcije CNV kod pacijenata upućenih na molekularnu kariotipizaciju. DNK pacijenata je analizirana na prisustvo CNV pomoću Agilent oligonukleotidnih platformi za komparativnu genomsku hibridizaciju na mikročipu. U našoj kohorti od 1412 pacijenata najveći broj je bio u pedijatrijskom uzrastu a odnos muškog i ženskog pola bio je 1,6: 1. Većina je imala NRP sa komorbiditetima. Iako su brojčano bile manje zastupljene, kod devojčica je bila značajno viša stopa detekcije klinički značajnih CNV ( dijagnostički doprinos): 20,3% (110/542) u poređenju sa 13,4% kod dečaka (117/870); p=7x10-4. Devojčice su imale složeniji fenotip (veći broj komorbiditeta), što bi moglo biti objašnjenje ovakvog nalaza, ali razlika u dijagnostičkom doprinosu kod devojčica i dečaka je bila značajna samo u grupama sa ≤2 komorbiditeta. Prevaga patogenih CNV kod devojčica je bila posebno izražena u podgrupama sa kongenitalnim srčanim manama, urogenitalnim anomalijama i poremećajima iz autističnog spektra. Sva tri entiteta su u opštoj populaciji češća kod muškog pola, što ukazuje na viši prag osetljivosti na ove poremećaje kod osoba ženskog pola. Veća zastupljenost patogenih CNV kod devojčica, potrebna za ispoljavanje istog fenotipa, podržava hipotezu o protektivnom efektu ženskog pola.