Analysis of Cytogenetic Abnormalities in Iranian Patients with Syndromic Autism Spectrum Disorder: A Case Series.

Objective: Autism spectrum disorder (ASD) is a heterogeneous neuropsychiatric group of pervasive developmental disorders mainly diagnosed through the complex behavioral phenotype. According to strong genetic involvement, detecting the chromosome regions and the key genes linked to autism can help to elucidate its etiology. The present study aimed to investigate the value of cytogenetic analysis in syndromic autism and find an association between autism and chromosome abnormalities. Materials & Methods: Thirty-six autistic patients from 30 families were recruited, clinically diagnosed with the Diagnostic and Statistical Manual of Mental Disorders (5th ed.; DSM-5). The syndromic patients with additional clinical features (including development delay, attention deficit, hyperactivity disorder, seizure, and language and intellectual impairment) were selected due to elevating the detection rate. Cytogenetics analysis was performed using GTG banding on the patients' cultured fibroblasts. Moreover, array-comparative genomic hybridization (CGH) was also performed for patients with a de novo and novel variant. Results: Karyotype analysis in 36 syndromic autistic patients detected chromosomal abnormalities in 2 (5.6%) families, including 46,XY,dup(15)(q11.1q11.2) and 46,XX,ins(7)(q11.1q21.3)dn. In the latter, array-CGH detected 3 abnormalities on chromosome 7, including deletion and insertion on both arms: 46,XX,del(7)(q21.11q21.3),dup(7)(p11.2p14.1p12.3)dn. Conclusion: We reported a novel and de novo cytogenetic abnormality on chromosome 7 in an Iranian patient diagnosed with syndromic autism. However, the detection rate in syndromic autism was low, implying that it cannot be utilized as the only diagnostic procedure.

P21-Associated ncRNA DNA Damage-Activated Expression in Bladder Cancer.

Úvod: Dlouhé nekódující ribonukleové kyseliny (long non-coding ribonucleic acids - lncRNA) jsou v poslední době vzhledem ke své úloze v procesu karcinogeneze předmětem zkoumání vědců zabývajících se nádory. Tyto transkripty regulují kritické kroky v normálních buněčných procesech, takže dysregulace jejich exprese se účastní patogeneze karcinomů. Z důvodu své blízkosti k lokusu CDKN1A má ncRNA spojená s P21 aktivovaná poškozením DNA (P21-associated ncRNA DNA damage activated - PANDA) v tomto ohledu zvláštní pozici. Podílí se na regulaci reakce na poškození DNA, stárnutí buněk a proliferace. Materiály a metody: V této studii jsme metodou kvantitativní polymerázové řetězové reakce hodnotili expresi této lncRNA ve tkáních karcinomu močového měchýře, sousedních nerakovinných tkání (adjacent non-cancerous tissues - ANCT) a v normálních vzorcích močového měchýře. Výsledky: Nebyl detekován žádný významný rozdíl v expresi PANDA, a to ani mezi nádorovými tkáněmi a ANCT (poměr exprese = 1,75; p = 0,11) nebo mezi nádorovými tkáněmi a normálními tkáněmi (poměr exprese = 2,72; p = 0,57). Úroveň exprese této lncRNA nebyla spojena s žádnými demografickými ani klinickými údaji o pacientech, jako je grade nádoru nebo recidiva, ani s rizikovými faktory souvisejícími s rakovinou, mezi něž patří např. kouření cigaret nebo závislost na opiu. Závěr: Tato studie tedy naznačuje, že PANDA není zapojena do patogeneze karcinomu močového měchýře. Hodnocení exprese jiných lncRNA by mohlo pomoci při identifikaci biomarkerů pro tyto karcinomy.

Sensitive Monogenic Noninvasive Prenatal Diagnosis by Targeted Haplotyping.

During pregnancy, cell-free DNA (cfDNA) in maternal blood encompasses a small percentage of cell-free fetal DNA (cffDNA), an easily accessible source for determination of fetal disease status in risk families through non-invasive procedures. In case of monogenic heritable disease, background maternal cfDNA prohibits direct observation of the maternally inherited allele. Non-invasive prenatal diagnostics (NIPD) of monogenic diseases therefore relies on parental haplotyping and statistical assessment of inherited alleles from cffDNA, techniques currently unavailable for routine clinical practice. Here, we present monogenic NIPD (MG-NIPD), which requires a blood sample from both parents, for targeted locus amplification (TLA)-based phasing of heterozygous variants selectively at a gene of interest. Capture probes-based targeted sequencing of cfDNA from the pregnant mother and a tailored statistical analysis enables predicting fetal gene inheritance. MG-NIPD was validated for 18 pregnancies, focusing on CFTR, CYP21A2, and HBB. In all cases we could predict the inherited alleles with >98% confidence, even at relatively early stages (8 weeks) of pregnancy. This prediction and the accuracy of parental haplotyping was confirmed by sequencing of fetal material obtained by parallel invasive procedures. MG-NIPD is a robust method that requires standard instrumentation and can be implemented in any clinic to provide families carrying a severe monogenic disease with a prenatal diagnostic test based on a simple blood draw.

FGFR2, FGF8, FGF10 and BMP7 as candidate genes for hypospadias.

Hypospadias is a common malformation, which results from failure of urethral tube closure, and whose molecular mechanisms are still largely unknown. The normal genital development is orchestrated by the urethral plate epithelium (UPE), at the genital tubercle (GT), which has polarizing activity, controlling a network of epithelial-mesenchymal interactions, which, when disturbed, may lead to hypospadias. Homeobox proteins (HOXs), fibroblast growth factors (FGFs) and bone morphogenic proteins (BMPs) are essential in this process. Hypospadias in the Hoxa13 -/- mice occurs as a result of the combined loss of Fgf8 and Bmp7 expression in the UPE. In both Fgf10 and Fgfr2 deficient mutant hypospadic male mice, cell proliferation is arrested prematurely and the maturation of the urethral epithelium is disrupted. Fgf8, Fgf10, and their receptor Fgfr2 are downstream targets of androgens (AR) during external genital development, an important fact given the pivotal role of AR in male sex differentiation. Therefore, we examined FGFR2, FGF10, FGF8, and BMP7 as candidate genes for hypospadias. DNA from 60 boys with familial, isolated, hypospadias was screened for mutations in FGFR2, FGF10, FGF8, and BMP7 genes, using DHPLC and DNA sequence analysis. The sequence variations c.590C>G and c.582-62G>A in FGF8, and, c.550+27C>T, c.727+180T>G, c.830T>C (p.Me186Thr), and c.2454C>T in FGFR2 were found uniquely in patients with hypospadias, as compared with 96 controls. No genetic variant in the other genes was detected. These results indicate that mutations are rare in FGF8 and FGFR2 in hypospadias, but gene variants may influence the risk.

Association of adenomatous polyposis coli (APC) gene polymorphisms with autism spectrum disorder (ASD).

We serendipitously identified a single nucleotide polymorphism (SNP), 8636C>A (rs1804197) in the 3'-untranslated region of the adenomatous polyposis coli (APC) gene to be associated with autism spectrum disorder (ASD). In order to gain further evidence for the association between the APC locus and ASD, we genotyped four additional adjacent common SNPs (rs2229992, rs42427, rs459552, and rs465899) in the coding regions within the APC gene in a set of Swedish ASDs and controls. One common haplotype TGAG was found to be associated with ASD after haplotype analysis using both Haploview v3.1.1 (P = 0.006) and COCAPHASE v2.403 (P = 0.030). This result is the first to suggest that the genomic locus at APC is associated with ASD, and that the APC gene itself is a good predisposing candidate to be evaluated in future studies due to its important role in neuronal development and function.

Cytokine single nucleotide polymorphisms in Iran.

Overall expression and secretion of cytokines are dependent on genetic nucleotide variations within or adjacent to regulatory regions of cytokine genes. This study allows the comparison of the prevalence of particular genetic markers. In 40 Iranian healthy subjects, cytokine single nucleotide polymorphisms (SNPs) were used to determine allelic and genotypic frequencies for the following cytokine genes: interleukin-1alpha (IL-1alpha) (T/C -889), IL-1beta (C/T -511, T/C +3962), IL-12 ( C/A-1188), interferon-gamma (IFN-gamma) (A/T UTR 5644), transforming growth factor-beta (TGF-beta) (C/T codon 10, G/C codon 25), tumor necrosis factor alpha (TNF-alpha) (G/A -308, G/A -238), IL-2 (T/G -330, G/T +166), IL-4 (T/G -1089, T/C -590, T/C -33), IL-6 (G/C -174, G/A nt565), IL-10 (G/A -1082, C/T -819, C/A -592), IL-1R (C/T pst11970), IL-1RA (T/C mspa 111100), IL-4RA (G/A +1902). All typing was performed using the PCR-SSP assay. Iranian and Italian, English, German, and Greek populations had similar cytokine profiles, but in some cases, the Iranian allele and genotype frequencies were significantly different from those of other Asian and African American populations for the majority of polymorphisms.