Путь пациента с редким диагнозом: нормативные документы и организация лечебно-диагностического процесса при орфанном заболевании в Российской Федерации

The main legislative document of the organization of medical care in the Russian Federation “On fundamental healthcare principles in the Russian Federation” and points related to the rare (orphan) diseases are  discussed. The organization of care, rules for managing a federal  registry of orphan diseases and routing of patients with main orphan nosological forms for which treatment is known are presented.Представлены правовые основы медицинской помощи лицам с редкими (орфанными) заболеваниями, используемые в Федеральном законе «Об основах охраны здоровья граждан в  Российской Федерации», правила ведения федерального регистра орфанных болезней и маршрутизация пациента с теми нозологическими формами, для которых предложено лечение

Chromosome instability, aging and brain diseases

Chromosome instability (CIN) has been repeatedly associated with aging and progeroid phenotypes. Moreover, brain‐specific CIN seems to be an important element of pathogenic cascades leading to neurodegeneration in late adulthood. Alternatively, CIN and aneuploidy (chromosomal loss/gain) syndromes exhibit accelerated aging phenotype

Somatic mosaicism in the diseased brain

It is hard to believe that all the cells of a human brain share identical genomes. Indeed, single cell genetic studies have demonstrated intercellular genomic variability in the normal and diseased brai

Комплексный анализ родства представителей подсемейства Cryptogrammoideae (Pteridaceae)

This research is the first comprehensive analysis of the intrageneric relationships inside the subfamily Cryptogrammoideae: 14 taxa of Coniogramme and one species of Cryptogramma were involved additionally in the molecular phylogenetic studies based on rbcL gene of plastid DNA; spore morphology of 32 taxa of cryptogrammoid ferns, namely 22 taxa of Coniogramme, nine species of Cryptogramma and one species of Llavea were studied using scanning electronic microscopy (SEM); 31 taxon of Cryptogrammoideae were studied using herbarium data from Herbaria across Europe and Asia (P, PE, LE, VLA, ALTB, TK) according to global botanical and geographical zones. As a result of this comprehensive analysis, we established a deep divergence of Coniogramme merillii in Coniogramme superclade: this species is the sister lineage to the remainder of Coniogramme. We revealed also the separateness of Co. suprapilosa from Co. rosthornii and Co. longissima, Co. africana from Co. lanceolata and Co. fraxinea, Co. robusta from Co. jinggangshanensis, Co. wilsonii and Co. japonica. Among Cryptogramma species, the relationship of Far Eastern Cr. gorovoi with Cr. crispa from the Caucasus and the Turkish endemic Cr. bithynica but not with any Far Eastern species was revealed. Spores of Coniogramme are characterized by simple smooth, granulate and papillate macroornamentation, spores of Cryptogramma species have the more coarse colliculate or tuberculate macro-ornamentation. Peculiarities of macro-ornamentation allow us to define six spore types in cryptogrammoid ferns: four spore types in Coniogramme and two spore types in Cryptogramma; the same spore type we assigned for Llavea cordifolia and Coniogramme suprapilosa. In Coniogramme, the grouping of species attending the spore type does not agree with existing classification and phylogenetic hypotheses. Genetic separateness of Co. suprapilosa corresponds with its exceptional verrucate spore sculpture not found in other Coniogramme species. In Cryptogramma, the grouping on the spore types corresponds with other morphological characteristics, existing system and molecular phylogeny. Spore ornamentation has diagnostic value in the recognition of cryptogrammoid taxa at the generic and section (in Cryptogramma) level

The Mechanisms of Resistance to Tyrosine Kinase Inhibitor Imatinib Therapy of Chronic Myeloid Leukemia

Imatinib mesylate is a potent and high selective inhibitor of Bcr-Abl tyrosine kinase, which is established now as the standard of Philadelphia chromosome positive (Ph) chronic myeloid leukemia (CML) treatment. The treatment of patients with chronic phase of CML with imatinib has resulted in high rates of hematologic and cytogenetic responses. Nevertheless, primary and acquired resistance have been observed in few CML patients. The mechanisms of resistance to imatinib and its clinical significance were discussed in this review

Наследственные заболевания легких и современные возможности генетической диагностики

The European Respiratory Society website gives the following criterion for the disease to be classified as rare (orphan) - the disease occurs in 1 person per 2 000. One of the well-studied rare lung diseases is cystic fibrosis (CF), which is often considered a medical care model for patients with other orphan diseases. However, effective diagnostics and therapies have not yet been developed for many other rare diseases. Moreover, their true prevalence remains unknown because these diseases often go undiagnosed. One of the problems in diagnosing rare diseases is the lack of knowledge among physicians.The aim of this review is to provide a brief clinical and genetic description of rare hereditary lung diseases and to show modern genetic diagnostics to raise awareness among physicians. Data from 95 articles on hereditary lung diseases were used.Results. The results of the analysis of lung diseases associated with bronchiectasis, fibrosis, pneumothorax, and hereditary storage diseases are presented. Genetics and diagnostics, including the three-step molecular genetic testing for cystic fibrosis, are considered in detail. The diagnosis has been developed for both neonatal screening and clinical manifestations. The emergence of targeted therapy based on genetic diagnosis makes neonatal screening even more relevant and leads to an increase in life expectancy. A patient registry was established within 10 years. A detailed analysis of the diagnosis of primary ciliary dyskinesia (PCD) is given, taking into account the absence of a single “golden” standard for the diagnosis of PCD. The genetic basis of the most common hereditary diseases and modern possibilities of their diagnosis are discussed, including sequencing of genes responsible for the development of orphan diseases using standard Sanger sequencing methods and next-generation sequencing, and creating multigene panels.Conclusion. New molecular diagnostic methods will help to understand the nature of orphan lung diseases, study their epidemiology, and develop new diagnostic algorithms. The study of the genetic causes of rare diseases may serve as a basis for the development of targeted therapy.На сайте Европейского респираторного общества (https://europeanlung.org/en/information-hub/factsheets/rare-and-orphan-lung-diseases/) приводится принятый в Европейском Союзе следующий критерий отнесения заболевания к редкому (орфанному): заболевание должно встречаться в популяции с частотой < 1 человека на 2 000 населения. Одним из хорошо изученных редких заболеваний легких является муковисцидоз (МВ) (кистозный фиброз), зачастую рассматриваемый как модель оказания медицинской помощи пациентам с другими орфанными заболеваниями. Однако в отличие от МВ, для многих других редких заболеваний действенные подходы к их раннему выявлению и эффективной терапии не разработаны. Более того, их истинная частота в популяции остается неизвестной, т. к. эти заболевания часто вообще не диагностируются. Одной из проблем диагностики редких заболеваний является недостаточная информированность врачей об этих болезнях.Целью обзора явилось описание краткой клинико-генетической характеристики редких наследственных заболеваний легких, оценка современных возможностей их генетической диагностики, а также повышение осведомленности врачей. Использовались данные 95 статей по наследственным заболеваниям легких.Результаты. Приведены результаты анализа заболеваний легких, сопровождающихся бронхоэктазами, фиброзом, пневмотораксом и наследственными болезнями накопления. Подробно рассмотрены генетика, диагностика, в т. ч. трехэтапное молекулярно-генетическое тестирование при МВ. Диагностика заболевания разработана как при неонатальном скрининге, так и по клиническим проявлениям. Появление таргетной терапии, основанной на генетическом диагнозе, делает неонатальный скрининг еще более актуальным, при этом продолжительность жизни пациентов увеличивается. Регистр пациентов создается в течение 10 лет. Приводится подробный анализ диагностики первичной цилиарной дискинезии (ПЦД) с учетом отсутствия единого метода — «золотого стандарта» диагностики ПЦД, как при МВ. Обсуждаются генетические основы наиболее частых наследственных заболеваний и современные возможности их диагностики (секвенирование генов, ответственных за развитие орфанных заболеваний, с использованием стандартных методов секвенирования по Сэнгеру и секвенирование нового поколения, создание мультигенных панелей).Заключение. Новые молекулярно-диагностические методы помогут не только понять природу орфанных заболеваний легких, но и позволят изучить их эпидемиологию и создать новые диагностические алгоритмы. Исследование генетических причин редких заболеваний имеет фундаментальное значение, т. к. может служить основой для разработки таргетной терапии

ROS-Induced DNA Damage Associates with Abundance of Mitochondrial DNA in White Blood Cells of the Untreated Schizophrenic Patients

Objective. The aim of this study was (1) to examine the leukocyte mtDNA copy number (CN) in unmedicated (SZ (m−)) and medicated (SZ (m+)) male patients with paranoid schizophrenia (SZ) in comparison with the healthy male controls (HC) and (2) to compare the leukocyte mtDNA CN with the content of an oxidation marker 8-oxodG in lymphocytes of the SZ (m−) patients. Methods. We evaluated leukocyte mtDNA CN of 110 subjects with SZ in comparison with 60 male HC by the method qPCR (ratio mtDNA/nDNA (gene B2M) was detected). SZ patients were divided into two subgroups. The patients of the subgroups SZ (m+) (N=55) were treated with standard antipsychotic medications in the hospital. The patients of the subgroup SZ (m−) (N=55) were not treated before venous blood was sampled. To evaluate oxidative DNA damage, we quantified the levels of 8-oxodG in lymphocytes (flow cytometry) of SZ (m−) patients (N=55) and HC (N=30). Results. The leukocyte mtDNA CN showed no significant difference in SZ (m+) patients and HC. The mtDNA CN in the unmedicated subgroup SZ (m−) was significantly higher than that in the SZ (m+) subgroup or in HC group. The level of 8-oxodG in the subgroup SZ (m−) was significantly higher than that in HC group. Conclusion. The leukocytes of the unmedicated SZ male patients with acute psychosis contain more mtDNA than the leukocytes of the male SZ patients treated with antipsychotic medications or the healthy controls. MtDNA content positively correlates with the level of 8-oxodG in the unmedicated SZ patients

Ribosomal DNA as DAMPs Signal for MCF7 Cancer Cells

Introduction: The cell free ribosomal DNA (cf-rDNA) is accrued in the total pool of cell free DNA (cfDNA) in some non-cancer diseases and demonstrates DAMPs characteristics. The major research questions: (1) How does cell free rDNA content change in breast cancer; (2) What type of response in the MCF7 breast cancer cells is caused by cf-rDNA; and (3) What type of DNA sensors (TLR9 or AIM2) is stimulated in MCF7 in response to the action of cf-rDNA?Materials and Methods: CfDNA and gDNA were isolated from the blood plasma and the cells derived from 38 breast cancer patients and 20 healthy female controls. The rDNA content in DNA was determined using non-radioactive quantitative hybridization. In order to explore the rDNA influence on MCF7 breast cancer cells, the model constructs (GC-DNAs) were applied: pBR322-rDNA plasmid (rDNA inset 5836 bp long) and pBR322 vector. ROS generation, DNA damage, cell cycle, expression of TLR9, AIM2, NF-kB, STAT3, and RNA for 44 genes affecting the cancer cell viability were evaluated. The methods used: RT-qPCR, fluorescent microscopy, immunoassay, flow cytometry, and siRNA technology.Results: The ratio R = cf-rDNA/g-rDNA for the cases was higher than for the controls (median 3.4 vs. 0.8, p < 10−8). In MCF7, GC-DNAs induce a ROS burst, DNA damage response, and augmentation of NF-kB and STAT3 activity. The number of the apoptotic cells decreases, while the number of cells with an instable genome (G2/M– arrest, micronuclei) increase. Expression of anti-apoptotic genes (BCL2, BCL2A1, BCL2L1, BIRC3, MDM2) is elevated, while expression of pro-apoptotic genes (BAX, BID, BAD, PMAIP1, BBC3) is lowered. The cells response for pBR322-rDNA is much more intense and develops much faster, than response for pBR322, and is realized through activation of TLR9- MyD88 - NF-kB- signaling. This difference in response speed is owing to the heightened oxidability of pBR322-rDNA and better ability to penetrate the cell. Induction of TLR9 expression in MCF7 is followed by blocking AIM2 expression.Conclusion: (1) Ribosomal DNA accumulates in cfDNA of breast cancer patients; (2) Cell free rDNA induce DNA damage response and stimulates cells survival, including cells with an instable genome; (3) Cell free rDNA triggers TLR9- MyD88- NF-kB- signaling, with significantly repressing the expression of AIM2