Seed size and cold stratification affect Acer negundo and Acer ginnala seeds germination

The aim of this work is to determine how the germination of seeds of the invasive tree Acer negundo depends on the period of cold stratification under the snow and the duration of stratification in the air on the branches of the trees. For comparison with A. negundo, we used seeds of Acer ginnala, introduced but not invasive tree in the Middle Urals. The period of stratification in the air modeled by collecting seeds in October and December. The duration of cold stratification under the snow was 0, 1, 2, 3 and 4 months. We hypothesized that the duration of stratification in the air did not affect the germination of A. negundo and A. ginnala seeds. Cold stratification under the snow had a positive effect on seed germination of both species. The best seed germination of A. negundo and A. ginnala was after 4 months of cold stratification under the snow, the germination rate differs: in A. negundo 12 ± 4% (small seeds) and 79 ± 7% (large seeds), in A. ginnala – 1 ± 2% (small seeds) and 18 ± 4% (large seeds). In both species, large seeds germinated at 7 to 18 times more intensively than small ones. In A. ginnala case, even after cold stratification under snow for 4 months, no more than 22% of the seeds germinated. The germination of A. ginnala seeds was 4–5 times lower than that of A. negundo seeds

Comparison of systemic and localized carrier-mediated delivery of methylprednisolone succinate for treatment of acute spinal cord injury

Localized carrier-mediated administration of drugs is a promising approach to treatment of acute phase of spinal cord injury (SCI) as it allows enhanced and/or sustained drug delivery to damaged tissues along with minimization of systemic side effects. We studied the effect of locally applied self-assembling micellar formulation of methylprednisolone succinate (MPS) with trifunctional block copolymer of ethylene oxide and propylene oxide (TBC) on functional recovery and tissue drug content after SCI in rats in comparison with local and systemic administration of MPS alone. Variations in the amplitude of motor evoked responses in the hindlimb muscles induced by epidural stimulation during acute phase of SCI and restoration of movements during chronic period after local vs. systemic application of MPS were evaluated in this study. Results demonstrate that local delivery of MPS in combination with TBC facilitates spinal cord sensorimotor circuitry, increasing the excitability. In addition, this formulation was found to be more effective in improvement of locomotion after SCI compared to systemic administration. LC–MS/MS data shows that the use of TBC carrier increases the glucocorticoid content in treated spinal cord by more than four times over other modes of treatment. The results of this study demonstrate that the local treatment of acute SCI with MPS in the form of mixed micelles with TBC can provide improved therapeutic outcome by promoting drug accumulation and functional restoration of the spinal cord

Infectious and non-infectious pericarditis in children

Pericardial diseases in children are heterogeneous in nature and can be both isolated and part of the systemic pathology. Data on the epidemiology and etiology of pericardial disease are contradictory and depend on the hospital profile, patients' age and study aims. Objective of the research-to study modern structure of pericardial diseases in children, clinical and instrumental features of individual forms and treatment tactics in real clinical practice according to the data of the Moscow multi-profile hospital. Study materials and methods: A complex clinical and laboratory-based examination was conducted in 121 patients aged from 1 month to 18 years, admitted to Morozov Children's City Clinical Hospital in Moscow in 2001-2016 with pericardium diseases. Results: pericardium inflammatory lesions were diagnosed in 86% of children, 57% of patients had infectious pericarditis (bacterial and idiopathic). The most severe course was in cases of bacterial, neoplastic pericarditis and postpericardiotomy syndrome (PPTS). A common feature of the severe course was the accumulation of a large pericardial effusion and the threat of a cardiac tamponade. In patients with idiopathic pericarditis and PPTS, herpesvirus infections markers, Mycoplasma pneumoniae, Chlamydophila pneumoniae, were more often detected with large effusions accumulation (p=0,02). Complications development was noted in 33 (27,3%) children: cardiac tamponade or the threat of its development in 23 (19%), recurrent course in 11 (9,1%). As anti-inflammatory therapy non-steroidal anti-inflammatory drugs were used (73,6% of patients); if they were inefficient-glucocorticosteroids (41,3%) and intravenous immunoglobulins (24,8%). Pericardiocentesis due to threat of cardiac tamponade was performed in 13 (10,74%) children. Conclusion: in pericarditis structure dominated infectious: bacterial and idiopathic (57%). Specific IgM antibodies to herpesviruses, Mycoplasma pneumoniae, Chlamydophila pneumoniae are possible markers of large pericardial effusion accumulation children with idiopathic pericarditis and PPTS. To assess the predictors of pericarditis adverse course incl. use of glucocorticosteroids, it is necessary to analyze long-term disease catamnesis. © 2017, Pediatria Ltd. All rights reserved

Infectious and non-infectious pericarditis in children

Pericardial diseases in children are heterogeneous in nature and can be both isolated and part of the systemic pathology. Data on the epidemiology and etiology of pericardial disease are contradictory and depend on the hospital profile, patients' age and study aims. Objective of the research-to study modern structure of pericardial diseases in children, clinical and instrumental features of individual forms and treatment tactics in real clinical practice according to the data of the Moscow multi-profile hospital. Study materials and methods: A complex clinical and laboratory-based examination was conducted in 121 patients aged from 1 month to 18 years, admitted to Morozov Children's City Clinical Hospital in Moscow in 2001-2016 with pericardium diseases. Results: pericardium inflammatory lesions were diagnosed in 86% of children, 57% of patients had infectious pericarditis (bacterial and idiopathic). The most severe course was in cases of bacterial, neoplastic pericarditis and postpericardiotomy syndrome (PPTS). A common feature of the severe course was the accumulation of a large pericardial effusion and the threat of a cardiac tamponade. In patients with idiopathic pericarditis and PPTS, herpesvirus infections markers, Mycoplasma pneumoniae, Chlamydophila pneumoniae, were more often detected with large effusions accumulation (p=0,02). Complications development was noted in 33 (27,3%) children: cardiac tamponade or the threat of its development in 23 (19%), recurrent course in 11 (9,1%). As anti-inflammatory therapy non-steroidal anti-inflammatory drugs were used (73,6% of patients); if they were inefficient-glucocorticosteroids (41,3%) and intravenous immunoglobulins (24,8%). Pericardiocentesis due to threat of cardiac tamponade was performed in 13 (10,74%) children. Conclusion: in pericarditis structure dominated infectious: bacterial and idiopathic (57%). Specific IgM antibodies to herpesviruses, Mycoplasma pneumoniae, Chlamydophila pneumoniae are possible markers of large pericardial effusion accumulation children with idiopathic pericarditis and PPTS. To assess the predictors of pericarditis adverse course incl. use of glucocorticosteroids, it is necessary to analyze long-term disease catamnesis. © 2017, Pediatria Ltd. All rights reserved

Анализ мутаций в генах CDC27, CTBP2, HYDIN и KMT5A при каротидных параганглиомах

Carotid paragangliomas (CPGLs) are rare neuroendocrine tumors that arise from paraganglionic tissue of the carotid body localizing at the bifurcation of carotid artery. These tumors are slowly growing, but occasionally they become aggressive and metastatic. Surgical treatment remains high-risk and extremely challenging; radiation and chemotherapy are poorly effective. The study of molecular pathogenesis of CPGLs will allow developing novel therapeutic approaches and revealing biomarkers. Previously, we performed the exome sequencing of 52 CPGLs and estimated mutational load (ML). Paired histologically normal tissues or blood were unavailable, so potentially germline mutations were excluded from the analysis with strong filtering conditions using 1000 Genomes Project and ExAC databases. In this work, ten genes (ZNF717, CDC27, FRG2C, FAM104B, CTBP2, HLA-DRB1, HYDIN, KMT5A, MUC3A, and PRSS3) characterized by the highest level of mutational load were analyzed. Using several prediction algorithms (SIFT, PolyPhen-2, MutationTaster, and LRT), potentially pathogenic mutations were identified in four genes (CDC27, CTBP2, HYDIN, and KMT5A). Many of these mutations occurred in the majority of cases, and their mutation type was checked using exome sequencing data of blood prepared with the same exome enrichment kit that was used for preparation of exome libraries from CPGLs. The majority of the mutations were germline that can apparently be associated with annotation errors in 1000 Genomes Project and ExAC. However, part of the mutations identified in CDC27, CTBP2, HYDIN, and KMT5A remain potentially pathogenic, and there is a large body of data on the involvement of these genes in the formation and progression of other tumors. This allows considering CDC27, CTBP2, HYDIN, and KMT5A genes as potentially associated with CPGL pathogenesis and requires taking them into account in further investigations. Thus, there is a necessity to improve the methods for identification of cancer-associated genes as well as pathogenic mutations.Каротидные параганглиомы (КПГ) - редкие нейроэндокринные опухоли, которые развиваются из параганглионарной ткани каротидного тельца и располагаются в области бифуркации сонной артерии. Эти опухоли характеризуются медленным ростом, однако в ряде случаев наблюдается агрессивное течение заболевания и метастазирование. Операции по удалению каротидных параганглиом сопряжены с высоким риском осложнений, в то время как лучевая и химиотерапия малоэффективны. Изучение молекулярного патогенеза КПГ может способствовать разработке новых подходов к терапии и открытию биомаркеров. Ранее нами выполнено высокопроизводительное секвенирование экзома 52 архивных образцов КПГ, одной из задач которого была оценка мутационной нагрузки. Из-за отсутствия парных образцов гистологически нормальных тканей или крови потенциально герминальные мутации были исключены из выборки с использованием баз данных 1000 Genomes Project и ExAC при строгих параметрах фильтрации. В настоящем исследовании проанализированы десять генов (ZNF717, CDC27, FRG2C, FAM104B, CTBP2, HLA-DRB1, HYDIN, KMT5A, MUC3A и PRSS3), характеризующихся наиболее высокой мутационной нагрузкой. В четырех генах (CDC27, CTBP2, HYDIN и KMT5A) идентифицированы потенциально патогенные мутации, согласно нескольким предсказательным алгоритмам (SIFT, PolyPhen-2, MutationTaster и LRT). Многие выявленные мутации оказались представленными в большом количестве образцов выборки, что заставило проверить их соматический статус с использованием дополнительных данных секвенирования экзома крови, выполненного с таким же набором для экзомного обогащения, как и при анализе опухолей КПГ. Обнаружено, что значительное число выявленных потенциально патогенных мутаций являются герминальными, что, по-видимому, связано с наличием ошибок аннотации в базах данных 1000 Genomes Project и ExAC. Однако часть идентифицированных мутаций в генах CDC27, CTBP2, HYDIN и KMT5A остаются предположительно патогенными, кроме того, имеются многочисленные данные о вовлеченности этих генов в формирование и прогрессию других видов опухолей. Это позволяет считать гены CDC27, CTBP2, HYDIN и KMT5A потенциально связанными с патогенезом КПГ и требует обратить на них особое внимание в дальнейших исследованиях. Таким образом, необходимо совершенствовать методики для выявления онко-ассоциированных генов и патогенных мутаций

Анализ мутаций в генах CDC27, CTBP2, HYDIN и KMT5A при каротидных параганглиомах

Carotid paragangliomas (CPGLs) are rare neuroendocrine tumors that arise from paraganglionic tissue of the carotid body localizing at the bifurcation of carotid artery. These tumors are slowly growing, but occasionally they become aggressive and metastatic. Surgical treatment remains high-risk and extremely challenging; radiation and chemotherapy are poorly effective. The study of molecular pathogenesis of CPGLs will allow developing novel therapeutic approaches and revealing biomarkers. Previously, we performed the exome sequencing of 52 CPGLs and estimated mutational load (ML). Paired histologically normal tissues or blood were unavailable, so potentially germline mutations were excluded from the analysis with strong filtering conditions using 1000 Genomes Project and ExAC databases. In this work, ten genes (ZNF717, CDC27, FRG2C, FAM104B, CTBP2, HLA-DRB1, HYDIN, KMT5A, MUC3A, and PRSS3) characterized by the highest level of mutational load were analyzed. Using several prediction algorithms (SIFT, PolyPhen-2, MutationTaster, and LRT), potentially pathogenic mutations were identified in four genes (CDC27, CTBP2, HYDIN, and KMT5A). Many of these mutations occurred in the majority of cases, and their mutation type was checked using exome sequencing data of blood prepared with the same exome enrichment kit that was used for preparation of exome libraries from CPGLs. The majority of the mutations were germline that can apparently be associated with annotation errors in 1000 Genomes Project and ExAC. However, part of the mutations identified in CDC27, CTBP2, HYDIN, and KMT5A remain potentially pathogenic, and there is a large body of data on the involvement of these genes in the formation and progression of other tumors. This allows considering CDC27, CTBP2, HYDIN, and KMT5A genes as potentially associated with CPGL pathogenesis and requires taking them into account in further investigations. Thus, there is a necessity to improve the methods for identification of cancer-associated genes as well as pathogenic mutations.Каротидные параганглиомы (КПГ) - редкие нейроэндокринные опухоли, которые развиваются из параганглионарной ткани каротидного тельца и располагаются в области бифуркации сонной артерии. Эти опухоли характеризуются медленным ростом, однако в ряде случаев наблюдается агрессивное течение заболевания и метастазирование. Операции по удалению каротидных параганглиом сопряжены с высоким риском осложнений, в то время как лучевая и химиотерапия малоэффективны. Изучение молекулярного патогенеза КПГ может способствовать разработке новых подходов к терапии и открытию биомаркеров. Ранее нами выполнено высокопроизводительное секвенирование экзома 52 архивных образцов КПГ, одной из задач которого была оценка мутационной нагрузки. Из-за отсутствия парных образцов гистологически нормальных тканей или крови потенциально герминальные мутации были исключены из выборки с использованием баз данных 1000 Genomes Project и ExAC при строгих параметрах фильтрации. В настоящем исследовании проанализированы десять генов (ZNF717, CDC27, FRG2C, FAM104B, CTBP2, HLA-DRB1, HYDIN, KMT5A, MUC3A и PRSS3), характеризующихся наиболее высокой мутационной нагрузкой. В четырех генах (CDC27, CTBP2, HYDIN и KMT5A) идентифицированы потенциально патогенные мутации, согласно нескольким предсказательным алгоритмам (SIFT, PolyPhen-2, MutationTaster и LRT). Многие выявленные мутации оказались представленными в большом количестве образцов выборки, что заставило проверить их соматический статус с использованием дополнительных данных секвенирования экзома крови, выполненного с таким же набором для экзомного обогащения, как и при анализе опухолей КПГ. Обнаружено, что значительное число выявленных потенциально патогенных мутаций являются герминальными, что, по-видимому, связано с наличием ошибок аннотации в базах данных 1000 Genomes Project и ExAC. Однако часть идентифицированных мутаций в генах CDC27, CTBP2, HYDIN и KMT5A остаются предположительно патогенными, кроме того, имеются многочисленные данные о вовлеченности этих генов в формирование и прогрессию других видов опухолей. Это позволяет считать гены CDC27, CTBP2, HYDIN и KMT5A потенциально связанными с патогенезом КПГ и требует обратить на них особое внимание в дальнейших исследованиях. Таким образом, необходимо совершенствовать методики для выявления онко-ассоциированных генов и патогенных мутаций

Comparative lipidomic analysis of inflammatory mediators in the aqueous humor and tear fluid of humans and rabbits

Introduction: Ocular inflammation is a key pathogenic factor in most blindness-causing visual disorders. It can manifest in the aqueous humor (AH) and tear fluid (TF) as alterations in polyunsaturated fatty acids (PUFAs) and their metabolites, oxylipins, lipid mediators, which are biosynthesized via enzymatic pathways involving lipoxygenase, cyclooxygenase or cytochrome P450 monooxygenase and specifically regulate inflammation and resolution pathways. Objectives: This study aimed to establish the baseline patterns of PUFAs and oxylipins in AH and TF by their comprehensive lipidomic identification and profiling in humans in the absence of ocular inflammation and comparatively analyze these compounds in the eye liquids of rabbits, the species often employed in investigative ophthalmology. Methods: Ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used for qualitative and quantitative characterization of lipid compounds in the analyzed samples. Results: A total of 28 lipid compounds were identified, including phospholipid derivatives and PUFAs, as well as 22 oxylipins. Whereas the PUFAs included arachidonic, docosahexaenoic and eicosapentaenoic acids, the oxylipins were derived mainly from arachidonic, linoleic and α-linolenic acids. Remarkably, although the concentration of oxylipins in AH was lower compared to TF, these liquids showed pronounced similarity in their lipid profiles, which additionally exhibited noticeable interspecies concordance. Conclusion: The revealed correlations confirm the feasibility of rabbit models for investigating pathogenesis and trialing therapies of human eye disorders. The identified metabolite patterns suggest enzymatic mechanisms of oxylipin generation in AH and TF and might be used as a reference in ocular inflammation studies. © 2020, Springer Science+Business Media, LLC, part of Springer Nature