About 50% of patients with arrhythmogenic cardiomyopathy (ACM) carry a pathogenic or likely patho... more About 50% of patients with arrhythmogenic cardiomyopathy (ACM) carry a pathogenic or likely pathogenic mutation in the desmosomal genes. However, there is a significant number of patients without positive familial anamnesis. Therefore, the molecular reasons for ACM in these patients are frequently unknown and a genetic contribution might be underestimated. Here, we used a next-generation sequencing (NGS) approach and in addition single nucleotide polymor-phism (SNP) arrays for the genetic analysis of two independent index patients without familial medical history. Of note, this genetic strategy revealed a homozygous splice site mutation (DSG2–c.378+1G>T) in the first patient and a nonsense mutation (DSG2–p.L772X) in combination with a large deletion in DSG2 in the second one. In conclusion, a recessive inheritance pattern is likely for both cases, which might contribute to the hidden medical history in both families. This is the first report about these novel loss-of-function mut...
Cystic fibrosis (CF) is one of the most common monogenic diseases with an autosomal recessive inh... more Cystic fibrosis (CF) is one of the most common monogenic diseases with an autosomal recessive inheritance. Carrier screening leads to a reduction in the number of children born with CF disease. The aim of this study was to develop the custom panel for the diagnosis of heterozygous carriage of polymorphic variants in the CFTR gene and to establish their allelic frequencies (AF) in one of the Russian regions where ethnic Russians predominate. Patients and Methods: The diagnostic panel was designed on the basis of data from the register of CF patients in Russia for 2017 and validated on 22 blood samples of patients with previously genetically established CF. The study participants (n=642) for CF variants estimation were randomly selected from the population-based cohort study ESSE-Vologda. Genotypes were determined by real-time PCR on the QuantStudio 12K Flex Real-Time PCR System. Data processing was performed using the TaqMan Genotyper Software. Results: The proposed diagnostic panel allowed simultaneous analysis of 60 variants of the CFTR gene. A total of 23 carriers of the following variants were identified among 642 participants: F508del (rs113993960) with a frequency of 2.02%, L138ins (rs397508686) and 394delTT (rs121908769)-0.47%, CFTRdele2.3 (c.54-5940_273+10250del21080; p. S18Rfs*16)-0.31%, R117H (rs78655421), and G542X (rs113993959)-0.16%. The frequency of heterozygotes in the Russian population was 3.58% or 1:28 (CI95%: 2.28-5.33% by Clopper-Pearson exact method). Conclusion: High frequency of heterozygous CFTR variants carriers and availability of highly productive diagnostic panel for detection of CFTR variants suggest the prospect of carrier screening for some common CF variants among Russian population.
years. Only 55% and 63% of the cohort had admission and post-ACS admission lipid checked respecti... more years. Only 55% and 63% of the cohort had admission and post-ACS admission lipid checked respectively. 75% of the patients had Atorvastatin 80 mg prescribed for secondary prevention. Calculation of 40% reduction in Non-HDL-C was obtained for 104 patients, for whom data was available; of these, 37.5% (39) achieved this target, while 62.5% (65) did not. It was noted that patients (74%) who met this target were statin-naive. 63% of them were already on statin prior to ACS diagnosis. On multivariate regression analysis, cigarette smoking (p <0.98), obesity (p <0.59), age (p <0.98), failed to predict Non-HDL-C reduction compared to being statin-naive (p <0.002). Conclusions: Study shows inadequate adherence to NICE lipid guideline for secondary prevention in a high risk population to prevent further CV events. Moreover, an absolute target of Non-HDL-C may be of better practical significance as a focus of lipid modification response.
VI International Symposium on Edible Alliaceae, 2012
We have studied the physical organization of genes and non-coding DNA in two Allium species, A. c... more We have studied the physical organization of genes and non-coding DNA in two Allium species, A. cepa and A. fistulosum. Expressed sequence tags (ESTs) clones and polymerase chain reaction (PCR) products of gene fragments obtained with primers designed for the gene sequences available in the public GenBank were physically mapped onto chromosomes using a Tyr-FISH technique. The allinase and lacrymatory factor synthase (LFS) genes that encoded enzymes operating in one metabolic way were cloned sequences and physically mapped onto A. cepa and A. fistulosum chromosomes. The alliinase gene probe (1100 bp) hybridized to the distal region of the long arm of chromosome 4 of A. cepa. The LFS gene probe (550 bp) hybridized to the proximal region of the long arm of chromosome 5 in both species. Inter-simple sequence repeats (ISSRs) that are located between SSR loci were used for chromosomal location of microsatellites in A. fistulosum using common FISH. The chromosomal organization of the Tyl-copia group retrotransposons were investigated in A. fistulosum. Dispersed hybridization of the probe along the chromosome arms apart from telomeric ends was detected. Chromosomal distribution of DNA methylation pattern in A. fistulosum L. was studied using a specific antibodies against 5-methylcytosine (anti-5mC). Highly methylated distal regions in all chromosomes were found. Differences in the methylation level between corresponding regions of homologue chromosomes were shown. We describe recent progress in exploiting the ultrasensitive Tyr-FISH technique for development of visual gene maps for chromosomes of A. cepa and A. fistulosum. The results on the chromosomal location of individual genetic loci aided in assembling physical and genetic maps. We related the physical organization of expressed genes to the contrasting patterns of chiasma distribution and to the organization of repetitive DNA family and highly methylated DNA in these two Allium species.
This is the first report on the presence of Ty3/gypsy like retrotransposons in the centromeric re... more This is the first report on the presence of Ty3/gypsy like retrotransposons in the centromeric region of Allium cepa and Allium fistulosum. We identified the putative Ty3/gypsy centromeric retrotrans posons (CR) among the DNA sequences of A. cepa present in the NCBI database and evaluated their copy number in the genomes of A. cepa and A. fistulosum. The putative copy number of Ty3/gypsy CR constituted about 26000 for A. cepa and about 7000 for A. fistulosum. The chromosomal organization of Ty3/gypsy CR was analyzed with fluorescent in situ hybridization (FISH). Primers were designed using the sequence ET645811 of A. cepa (Genome Survey Sequence database), and a 300 bp product was obtained from genomic DNA of A. cepa and A. fistulosum. The PCR product displayed similarity to the reverse transcriptase of the CR Ty3/gypsy family and was used as a probe for FISH. Hybridization signals were mainly localized in the centromeric regions of A. cepa chromosomes. On the chromosomes of A. fistulosum, however, the hybridiza tion patterns were less specific. The evolution of the CR Ty3/gypsy in these two closely related Allium species is discussed.
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Papers by Anna Kiseleva