Dynamics in the activity of peroxidase and its isoforms in leaves of different apple cultivars

Background. Various approaches are used for identification of the most resistant fruit crop cultivars, including the analysis of different physiological and biochemical indicators. In Krasnodar Territory, Russia, one of the major stressors in summer is the hydrothermal stress. Drought and heat lead to an oxidative stress, as reactive oxygen species are produced in plant cells. Plants respond to oxidative damage by activating antioxidant enzymes, such as superoxide dismutase, catalase, and various peroxidases. Peroxidase is able to decompose hydrogen peroxide. Peroxidase activity was calculated under natural summertime changes in the hydrothermal pattern (control) and in simulated high-temperature conditions.Materials and methods. Three apple cultivars of Russian breeding, ‘Fortuna’, ‘Soyuz’ and ‘Prikubanskoe’, and cv. ‘Ligol’ of Polish origin were studied. In the summers of 2018–2019, their leaf samples were analyzed to assess peroxidase activity and its isozyme composition under control and stress conditions. Native electrophoresis in polyacrylamide gel was used for separation of peroxidase isoforms. Malondialdehyde content was measured to identify oxidative stress levels in apple leaves.Results. The tested indicators demonstrated a high degree of heterogeneity induced by both cultivar specificity and seasonal weather dynamics. Peroxidase isoforms with a molecular weight of 70 to 60 kDa, characterized by the maximum level of variability (1–4 isoforms), were isolated. Two other groups included 1–3 isoforms with a molecular weight of ~130–100 kDa, and one with a molecular weight of ~55 kDa. The highest enzyme activity was found in cvs. ‘Fortuna’ and ‘Soyuz’ in July 2018, the hottest month during the period of research. Under simulated conditions, the triploid cultivar ‘Soyuz’ was least susceptible to the stress impact

Evolution of Highly Polymorphic T Cell Populations in Siblings with the Wiskott-Aldrich Syndrome

Population level evolutionary processes can occur within a single organism when the germ line contains a mutation that confers a cost at the level of the cell. Here we describe how multiple compensatory mutations arose through a within-individual evolutionary process in two brothers with the immune deficiency Wiskott-Aldrich Syndrome (WAS). As a result, both brothers have T lymphocyte populations that are highly polymorphic at the locus of the germ line defect, and no single allele achieves fixation. WASP, the gene product affected in this disease, is specific to white blood cells where it is responsible for regulating actin cytoskeleton dynamics in a wide range of cellular responses. The brothers inherited a rare allele predicted to result in truncated WASP lacking the carboxy-terminal VCA domains, the region that directly catalyzes actin filament generation. Although the brothers' T cell populations are highly polymorphic, all share a corrective effect relative to the inherited allele in that they restore the VCA domain. This indicates massive selection against the truncated germ line allele. No single somatic allele becomes fixed in the circulating T cell population of either brother, indicating that a regulated step in maturation of the affected cell lineage is severely compromised by the germ line allele. Based on the finding of multiple somatic mutations, the known maturation pathway for T-lineage cells and the known defects of T cells and precursor thymocytes in mice with truncated WASP, we hypothesize that the presence of truncated WASP (WASPΔVCA) confers an extreme disadvantage in early developing thymocytes, above and beyond the known cost of absence of full-length WASP, and that the disadvantage likely occurs through dominant negative competition of WASPΔVCA with N-WASP, a protein that otherwise partially compensates for WASP absence in developing thymocytes

ИССЛЕДОВАНИЕ АНТИГЕН-СТИМУЛИРОВАННОЙ ПРОДУКЦИИ υ-ИНТЕРФЕРОНА EX VIVO В ПЕРИФЕРИЧЕСКОЙ КРОВИ У БОЛЬНЫХ АКТИВНЫМ ТУБЕРКУЛЕЗОМ ЛЕГКИХ

Tuberculosis (TB) is one of the most significant problems in the Russian Health Care. Russia remains on the list of the 22 countries with a high TB incidence and on the third place in the world with a high prevalence of Drug Resistant TB [1]. It is urgently needed to develop new TB diagnostic methods as well as effective measures of the specific TB prevention, including a development of the novel vaccines, so we have to know better about the most immunogenic antigens of Mycobacterium Tuberculosis. We studied the Interferon-Q production in the whole blood after stimulating immune response with different proteins of Mycobacterium Tuberculosis in patients with active TB. The study results permitted us to evaluate the immunogenicity of the previously known proteins (Ag85a и ESAT-6) in comparison to the recently identified ones (Rv2957, Rv2958c и Rv0447), analyzing simultaneously their relation to tuberculin, as well as to antigens of the different viruses (Human Immunodeficiency Virus, Cytomegalovirus, Epstein-Barr Virus, Influenza Virus). Protein Rv2958c, unlike protein ESAT-6, showed the high immunogenicity in comparison to tuberculin. The expressed immunogenicity of protein Rv2958c might be indicated a possible greatest specificity of immune response to this antigen in TB patients. Meanwhile, bacillary tuberculosis was strongly associated with low immune response to this protein. Also we were found statistical differences in immune responses of patients to the different Mycobacterium Tuberculosis antigens depending on the drug sensitivity. In addition it was interesting to know about a significantly low immune response of patients with Drug Resistant TB to protein pp65 CMV.Туберкулез является одной из наиболее серьезных проблем российского здравоохранения. Россия остается в списке из 22 стран с высокой заболеваемостью туберкулезом и на 3-м месте в мире по распространенности лекарственно-устойчивых форм заболевания. Требуется разработка как новых методов диагностики, так и эффективных мер специфической профилактики, включая новые вакцины, для создания которых необходимо знание наиболее иммуногенных антигенов Mycobacterium tuberculosis. В данной работе исследовалась продукция интерферона-гамма в цельной крови пациентов с активным туберкулезом в ответ на антиген-стимуляцию различными белками Mycobacterium tuberculosis. Результаты исследования позволили дать оценку иммуногенности ранее изученных белков (Ag85a и ESAT -6) в сравнении с недавно идентифицированными белками (Rv 2957, Rv 2958c и Rv0447) с одновременным изучением их отношений к туберкулину и антигенам различных вирусов (вирус иммунодефицита человека, цитомегаловирус, вирус Эпштейна – Барр, вирус гриппа). Белок Rv2958c, в отличие от белка ESAT T-6, показал большую иммуногенность при сравнении с туберкуэффективлином. Выраженная иммуногенность белка Rv2958c может свидетельствовать о возможно большей специфичности иммунного ответа на этот антиген у больных туберкулезом. Между тем бактериовыделение было ассоциировано с достоверно низким иммунным ответом на данный белок. Также выявлены статистические различия в иммунореактивности пациентов к различным антигенам Mycobacterium tuberculosis в зависимости от наличия или отсутствия лекарственной устойчивости возбудителя. Представляет интерес достоверно низкая иммунореактивность пациентов с лекарственно-устойчивым туберкулезом в отношении белка pp65 CMV

Ocrelizumab versus Interferon Beta-1a in Relapsing Multiple Sclerosis

Supported by F. Hoffmann–La Roche

DIAGNOSIS OF OCCIPITAL LOBE EPILEPSY

Objective. The study aimed to investigate the significance of MRI and video electro-encephalography (EEG) monitoring in diagnosis of occipital lobe epilepsy.Materials and methods. The study involved 103 patients aged 2-17 years old who were diagnosed with occipital lobe epilepsy. All patients underwent brain MRI scans using a CT scanner 1.5 TL in the T1 and T2 modes in the axial, sagittal and frontal planes. For video electro-encephalography monitoring the EEG-recorder Encephalan 9 was used.Results. For the analysis of MRI studies in patients with occipital epilepsy, we grouped the patients as follows: with signs of perinatal brain lesions – 39 patients (38%), with congenital abnormalities of brain development – 26 patients (25%), and those with no pathological findings in the brain structures – 38 (37%). The EEG monitoring results revealed no deviations from the normal background rhythm in 27 patients (26%). Fifty patients (49%) demonstrated abnormalities of the alpha rhythm. In 26 patients (25%) a slowdown of the background rhythm was detected. The results demonstrated that a slowdown and abnormalities of the background rhythm occurred more often in children with symptomatic occipital lobe epilepsy, while these EEG changes occurred the least in patients with Lennox-Gastaut Syndrome. All patients showed the epileptiform activities manifested by the “peak – slow wave” complexes similar to the benign epileptiform rhythm patterns.Conclusion. The results obtained from the brain MRI studies in patients with occipital lobe epilepsy indicate that the perinatal brain damage has a role in the pathogenesis of the disease. The EEG results show that the EEG patterns in Panayiotopoulos Syndrome are more variable as compared to Lennox-Gastaut Syndrome and symptomatic occipital lobe epilepsy

Окислительные процессы и антиоксидантные реакции у винограда при заражении милдью на фоне прайминга микроорганизмами

Priming plants with natural agents, including microorganisms, is a promising alternative to chemical methods of protection in growing plants and, particularly, in viticulture. However, the molecular mechanisms of the priming phenomenon are still not fully elucidated. The antioxidant system and reactive oxygen species are known to effectively modulate plant responses to various external influences. This study aimed to identify the relationship between the priming of grapes by microorganisms and the functioning of the antioxidant system in a protective response to downy mildew infection. The experiment was carried out on leaf discs of Muscat blanc susceptible to downy mildew infected with Plasmopara viticola and treated with microorganisms incompatible with the pathogen, as well as with the corresponding symbiotic microorganisms. During the compatible interaction between P. viticola and grapes, oxidative processes were suppressed with viniferin formation. Leaf treatments with Saccharomyces cerevisiae and Bacillus subtilis effectively curbed the development of downy mildew on grape leaves. Priming with these microorganisms did not lead to a significant change in the biochemical parameters of grapes. Nevertheless, subsequent downy mildew infection initiated the formation of viniferin and maintained H2O2 content at a high level. Thus, priming with microorganisms eliminates the physiological effects of compatible interactions between downy mildew and grapes associated with blocking oxidative processes. To suppress pathogen development, host defenses and antagonistic effects of microbial priming agents are requiredПрайминг растений естественными природными агентами, в том числе микроорганизмами, представляет интересную альтернативу химическим методам защиты в растениеводстве и в виноградарстве в частности. Однако молекулярные механизмы этого явления до конца не ясны. Известно, что антиоксидантная система растений и образование активных форм кислорода эффективно модулируют реакции растений на множество внешних воздействий, в том числе биотический стресс. В связи с этим цель исследования – выявление связи прайминга винограда микроорганизмами и функционирования элементов антиоксидантной системы в защитном ответе на заражение милдью. Эксперимент проводили на листовых дисках винограда восприимчивого к милдью сорта Мускат белый, зараженных Plasmopara viticola на фоне обработок несовместимыми и симбиотическими организмами. Определение содержания стильбенов, аскорбиновой кислоты проводили методом капиллярного электрофореза, содержание перекиси водорода, ТБК‑активных веществ и активность ферментов – спектрофотометрически. Совместимое взаимодействие между P. viticola и виноградом характеризовалось подавлением окислительных процессов и образованием микроботоксичной окисленной формы ресвератрола – виниферина. Эффективными в сдерживании развития милдью были обработки листьев Saccharomyces cerevisiae и Bacillus subtilis. Прайминг данными микроорганизмами не приводил к существенному изменению биохимических показателей винограда, однако последующее заражение инициировало образование виниферина и сохранение содержания H2O2 на высоком уровне. Таким образом, прайминг микроорганизмами нивелирует физиологические эффекты совместимого взаимодействия милдью и винограда, связанные с блокированием окислительных процессов у винограда. Для полного сдерживания развития патогена требуются защитные реакции растения-хозяина и антагонистическое воздействие со стороны микроорганизмов – агентов прайминг

Antigen induced production of υ-interferon ex vivo, in the peripheral blood of patients with active pulmonary tuberculosis

Tuberculosis (TB) is one of the most significant problems in the Russian Health Care. Russia remains on the list of the 22 countries with a high TB incidence and on the third place in the world with a high prevalence of Drug Resistant TB [1]. It is urgently needed to develop new TB diagnostic methods as well as effective measures of the specific TB prevention, including a development of the novel vaccines, so we have to know better about the most immunogenic antigens of Mycobacterium Tuberculosis. We studied the Interferon-Q production in the whole blood after stimulating immune response with different proteins of Mycobacterium Tuberculosis in patients with active TB. The study results permitted us to evaluate the immunogenicity of the previously known proteins (Ag85a и ESAT-6) in comparison to the recently identified ones (Rv2957, Rv2958c и Rv0447), analyzing simultaneously their relation to tuberculin, as well as to antigens of the different viruses (Human Immunodeficiency Virus, Cytomegalovirus, Epstein-Barr Virus, Influenza Virus). Protein Rv2958c, unlike protein ESAT-6, showed the high immunogenicity in comparison to tuberculin. The expressed immunogenicity of protein Rv2958c might be indicated a possible greatest specificity of immune response to this antigen in TB patients. Meanwhile, bacillary tuberculosis was strongly associated with low immune response to this protein. Also we were found statistical differences in immune responses of patients to the different Mycobacterium Tuberculosis antigens depending on the drug sensitivity. In addition it was interesting to know about a significantly low immune response of patients with Drug Resistant TB to protein pp65 CMV