ABSTRACT
Aim: To develop magnetic nanoparticles (MNPs) based on iron oxide for DNA isolation from blood cells for quantitative molecular genetic analyses of the V617F mutation in the Januskinase 2 (JAK2) gene. Materials and Methods: MNPs were synthesized by the coprecipitation method and coated with tetraethyl orthosilicate (TEOS). The size and shape of the complexes were estimated using transmission electron microscopy. Twenty blood samples from patients with myeloproliferative disorders were used for DNA isolation with the MNPs. DNA quality and compatibility for molecular genetic studies of the JAK2 V617F mutation were investigated by gel electrophoresis and real-time polymerase chain reaction (RT-PCR). Results: The average amount of DNA isolated from 150 µL of whole blood was 75.2 ng when MNPs were used and 72.5 ng when standard silica sorbent was used. There was no DNA damage observed after interaction with MNPs. RT-PCR demonstrated similar values for the JAK2 V617F mutant DNA ratios in the samples after DNA isolation with MNPs and by standard sorption on silica. Conclusions: MNPs with silicate capsules of sufficient thickness were obtained and the undesirable damaging effect of iron oxides on nucleic acids during isolation from cells were eliminated. Designed MNPs allow obtaining intact DNA for molecular genetic studies using the example of the JAK2 V617F for study.
Subject(s)
DNA/isolation & purification , Genetic Testing/methods , Hematologic Diseases/diagnosis , Magnetic Iron Oxide Nanoparticles/chemistry , Silicon Dioxide/chemistry , DNA/chemistry , DNA/genetics , DNA Mutational Analysis/methods , Hematologic Diseases/blood , Hematologic Diseases/genetics , Humans , Janus Kinase 2/genetics , MutationABSTRACT
Aim: The purpose of our study was to analyze the predictive ability of the multiplicative model of genetic risk of nonlacunar ischemic stroke (IS) for independent samples from Russia. Patients & methods: A total of 181 patients and 360 healthy controls were included in this study. The discriminative accuracy of model was evaluated by the area under the receiver operating characteristic curve (AUC). Results: Classification model based on 15 single-nucleotide polymorphisms (SNPs), which are associated with a cardioembolic subtype of IS, had an AUC of 0.62 in patients with corresponding subtypes and an AUC of 0.58 for all patients. Conclusion: Risk calculation approach based on IS-associated SNPs had satisfactory performance in predicting the predisposition to the disease.
Subject(s)
Brain Ischemia/genetics , Genetic Testing/methods , Stroke/genetics , Adult , Aged , Area Under Curve , Case-Control Studies , Female , Genetic Predisposition to Disease/genetics , Genotype , Humans , Male , Middle Aged , Polymorphism, Single Nucleotide/genetics , Precision Medicine/methods , ROC Curve , Risk Factors , RussiaSubject(s)
Blood Platelets/metabolism , Gene Expression Regulation, Neoplastic , Hematologic Neoplasms/metabolism , MicroRNAs/biosynthesis , Myeloproliferative Disorders/metabolism , RNA, Neoplasm/biosynthesis , Thrombocytosis/metabolism , Adult , Blood Platelets/pathology , Female , Hematologic Neoplasms/genetics , Hematologic Neoplasms/pathology , Humans , Male , MicroRNAs/genetics , Middle Aged , Myeloproliferative Disorders/genetics , Myeloproliferative Disorders/pathology , RNA, Neoplasm/genetics , Thrombocytosis/genetics , Thrombocytosis/pathologySubject(s)
Algorithms , Calreticulin/genetics , Fusion Proteins, bcr-abl/genetics , Janus Kinase 2/genetics , Leukemia, Myeloid, Acute/genetics , Myeloproliferative Disorders/genetics , Receptors, Thrombopoietin/genetics , Aged , Female , Fusion Proteins, bcr-abl/blood , Humans , Leukemia, Myeloid, Acute/blood , Male , Middle Aged , Mutation , Myeloproliferative Disorders/bloodSubject(s)
Adaptor Proteins, Signal Transducing/genetics , Myeloproliferative Disorders/genetics , Adult , Aged , Aged, 80 and over , Case-Control Studies , Female , Healthy Volunteers , Humans , Janus Kinase 2/genetics , Male , Middle Aged , Myeloproliferative Disorders/blood , Myeloproliferative Disorders/epidemiology , Polymorphism, Single Nucleotide , Young AdultABSTRACT
To study the molecular epidemiology of HIV-1 in Krasnoyarsk region, Russia, where HIV-1 has spread rapidly since 2000, we obtained pol sequences from individuals living in this region (n = 67) as well as in the geographically closely related Altay region (n = 13). In both regions, subtype A viruses specific for the former Soviet Union (IDU-A strains) were dominant (92.5%). Virus sequences clustered according to the geographic origin of the infected individuals rather than to their risk group, demonstrating the role of geographically defined epidemiological networks in the propagation of the HIV-1 epidemic in the region. Six viruses belonged to subtype B. Three of them were phylogenetically (and therefore epidemiologically) closely related to each other, demonstrating that even though IDU-A viruses dominate the epidemic, the spread of other virus strains does occur. Most viruses (75%) had an A62V mutation in reverse transcriptase, specific for HIV-1 strains in Russia. Remarkably, 26 of 47 (55%) patients under HAART with detectable virus loads did not have any known drug-resistant mutation, indicating the need to increase compliance to therapy.