Experimental application results of mesenchymal stem cell microvesicles in the mouse model of acute renal failure

An important role in restoration of damaged organs and tissues is played by mesenchymal stem cells (MSCs) and microvesicular particles (MV) produced by them. They can be a source of cytokines, anti- apoptotic and growth stimulating factors. In addition, MVs carry out transport of mRNA, miRNA, and signal proteins into damaged tissues. This increases the ability of cells to regenerate and to inhibit apoptosis, promote to angiogenesis and stimulate cell proliferation. The aim of our research was to study the immunoregulatory and pro-regenerative properties of mesenchymal stem cell microvesicles (MSC-MV) in a model of glycerol- induced acute renal failure (ARF) in mice. The experiments were carried out on CBA mice aged 3-4 months. AKI was induced by a single intramuscular injection of 50% glycerol. MSCs were obtained from the bone marrow of healthy animals and cultivated under standard conditions. Microvesicles were obtained by centrifugation at 12000g of MSC supernatant after induction of their apoptosis by culturing under oxygen deprivation conditions and in serum-free medium. MSC-MV was injected intravenously into the retroorbital sinus one day after induction of ARF. The MV dose was calculated as equivalent to (derived from) 1 million MSCs, which was 100 mL per mouse. Animals were taken out of the experiment on days 4 and 11 after MSC-MV injection. Blood plasma was taken to determine the level of creatinine, urine – for albumin analysis, kidneys – for histological examination. It has been shown that MVs induced by MSCs dose-dependently stimulated splenocyte proliferation in both spontaneous and Con-A induced tests. The addition of MV caused a decrease in doxorubicin-induced apoptosis of splenic lymphocytes in mice. Probably, in this case, MV produced by MSCs had an immunostimulatory and antiapoptotic effect. Also, MVs had a positive impact on the restoration of structure and function kidneys in a model of ARF in mice. The use of MSC-MV in treatment of acute renal failure induced by a single injection of 50% glycerol contributed to decrease albumin level urine and restoration of creatinine level in blood serum of animals. Morphological studies have shown decrease in the height cell and collecting duct diameter in the medulla and a decrease in the largest transverse diameter of superficial glomeruli in the renal cortex of sick mice. Thus, the obtained results indicate significant therapeutic and pro-regenerative properties of MSC-MV, which require further study

PHENOTYPIC CHARACTERIZATION AND INTRACELLULAR CYTOKINES OF MEMORY T-CELLS IN MULTIPLE SCLEROSIS PATIENTS AFTER T-CELL VACCINATION

Multiple sclerosis (RS) patients were treated with a vaccine consisted of autological mielin-reactive T-cells. Effector memory CD8+ CD45RO+CD62L- T-cells as well as central memory CD4+ CD45RO+CD62L+ and CD8+ CD45RO+CD62L+ T-cells were significantly increased in MS patients as compared with levels in healthy individuals. T-cells vaccination had no effect on the T-cells memory subpopulation composition. However, vaccine-treated RS patients had significant (by 2 fold) reduction of relative quantity of CD4+IFNγ+IL-4— and CD8+IFNγ+IL-4— memory T-cells, as well as suppression by 6 fold of CD4+ memory cells number, which produce both kinds of cytokines

Population immunity to SARS-CoV-2 virus in residents of the Irkutsk Region in the dynamics of the epidemic

Background. Currently, the COVID-19 pandemic in the world and in Russia remains the main event. In this regard, the study of the manifestations of the epidemic process of the new coronavirus infection COVID-19 and the patterns of its development are an urgent area of research. In the fight against this viral disease, an important role is assigned to the study of the development of population immunity to the SARSCoV-2 virus, which will make it possible to assess the dynamics of seroprevalence and the formation of post-infectious humoral immunity, forecasting the development of the epidemiological situation, elucidating the characteristics of the epidemic process, and will also contribute to planning activities for specific and non-specific prevention of the disease.The aim: to determine the dynamics of population immunity to SARS-CoV-2 among the population of the Irkutsk region during the COVID-19 pandemic.Materials and methods. As a part of the Rospotrebnadzor project of assessing population immunity to SARS-CoV-2 in the population of the Russian Federation, the research has being conducted among the population of the Irkutsk region in the periods from June 23, 2020 to July 19, 2020 (Stage 1), from September 16, 2020 to September 25, 2020 (Stage 2), from December 7, 2020 to December 18, 2020 (Stage 3) and from March 8, 2021 to March 14, 2021(Stage 4), taking into account the reacted one recommended by the WHO. The content of antibodies to SARS-CoV-2 was determined by ELISA using a set of tests for human serum or plasma for specific immunoglobulins of class G to the proteins of the SARS-CoV-2 coronavirus.The results. The research of the humoral immunity of volunteers shows that during the period of an epidemic rise in the incidence of COVID-19 in the Irkutsk region, a low level of seroprevalence was formed (Stage 1 – 5.8 ± 0.5 %, Stage 2 – 12.1 ± 0.7 %), and in conditions of a long-term maximum increase in the incidence rate – 25.9 ± 1.0 % (Stage 3) and 46.2 ± 1.2 % (Stage 4). A significant proportion (Stage 1 – 82.2 ± 3.2 %, Stage 2 – 86.1 ± 2.3 %) of asymptomatic forms of infection characterizes the high intensity of the latently developing epidemic process in the first two stages. High levels of IgG in reconvalescents of COVID-19 persisted for an average of 3 to 5 months.Conclusion. The results of assessing the population immunity to the SARS-CoV-2 virus in the population of the Irkutsk region indicate that the seroprevalence level at Stage 4 of the research was 46.2 %. After the disease, on average, 49.5 % of persons did not detect antibodies. The results obtained should be taken into account when organizing preventive measures, including vaccination, and predicting morbidity

ДИНАМИКА ВЫСШИХ ПСИХИЧЕСКИХ ФУНКЦИЙ У БОЛЬНЫХ СО СТЕНОЗИРУЮЩИМ ПОРАЖЕНИЕМ ВНУТРЕННИХ СОННЫХ АРТЕРИЙ ДО И ПОСЛЕ ХИРУРГИЧЕСКОЙ РЕВАСКУЛЯРИЗАЦИИ ГОЛОВНОГО МОЗГА

Obliterating atherosclerosis of internal carotid arteries is one of the main causes of ischemic stroke and discirculatory encephalopathy. It causes up to 40% of ischemic disorders of cerebral circulation. Currently, the strategy for stroke prevention is determined by the intensive development of surgical methods of treatment, primarily methods for managing lesions of brachiocephalic arteries. Based on the results of a number of international multicenter randomized studies, indications for reconstructive operations for BCA, tactics for managing patients in the postoperative period were formulated. A number of patients with atherosclerotic lesions of brachiocephalic arteries have reduced cognitive functions. The aim of the study is to compare cognitive functions (CF) in patients who underwent different surgical approaches in the treatment of obliterating atherosclerotic lesion of internal carotid arteries (ICA).MATERIAL AND METHODS. We studied higher mental functions (HMFs) in 116 patients with obliterating unilateral or bilateral lesion of ICA. The study of cognitive functions (MF) was performed prior to carotid endarterectomy (CE, group 1, n=73) and transluminal balloon angioplasty of ICAs (TBA of ICA, group 2, n=43), and on days 5–7 and 30–31 after cerebral revascularization (CR). To assess the overall severity of cognitive impairment, the summary indicators of main screening neuropsychological tests were used: MMSE; MoCA; Frontal Assessment Battery (FAB); Beck Depression Inventory and Hamilton Depression Rating Scale.RESULTS. Results Neuropsychologic disorders were reavealed in 98% of patients prior to surgery. An initially comparable condition of HMF in groups with CE and TBA of ICA was revealed. MMSE2 revealed a significant improvement in the results in group 1 both in comparison with the initial data (p<0.05) and in comparison with the results of the second test of group 2. The results of MMSE1 and MMSE2 in group 2 did not show significant differences (p>0.05). MMSE3 showed a significant improvement in the test results in each group compared to MMSE1 and MMSE2. MMSE3 results were significantly better (p<0.01) in group 1 than in group 2. MoCA revealed most significant differences in group 1 results with a significant decrease in cognitive dysfunction both in MoCA1, MoCA2, MoCA3 either within the group (p><0.05) or compared to MoCA3 data between CE group and TBA group (p><0.01). FAB showed that the FAB2 test amounted to 16.2 points in the group with CE and 14.6 points in the group with TBA of ICAs (p><0.05). A significant improvement in the performance of mnestic functions was noted when examining patients in the dynamics of FAB3: 17.3 and 15.6 points (p><0.05), respectively. According to Hamilton Depresion Rating Scale and Beck Depression Inventory, both groups of patients showed a moderate level of anxiety and depression (12.4 and 15.8 according to Hamilton Depression Rating Scale (p><0.05), 12.3 and 14.4 according to Beck Depression Inventory (p>0.05)). During the second test, the depressive mood of patients was reduced (on the Hamilton scale 8.4 and 13.8 points.CONCLUSION When comparing HMFs in patients who underwent a different surgical approach (CE vs TBA of ICAs) in the treatment of obliterating atherosclerotic lesions of ICAs, we found that: 1) the maximum improvement in HMFs appears by the 30th day of the postoperative period in comparison with preoperative parameters; 2) the most significant improvement HMFs test parameters by the 30th day of the postoperative period is noted in the group where CE was used as the method of surgical revascularization of the brain.ВВЕДЕНИЕ. Одной из основных причин ишемического инсульта и дисциркуляторной энцефалопатии является стенозирующий атеросклероз внутренних сонных артерий (ВСА). Он служит причиной до 40% случаев ишемических нарушений мозгового кровообращения. В настоящее время стратегию профилактики инсульта определяет интенсивное развитие хирургических методов лечения, прежде всего патологии брахиоцефальных артерий (БЦА). По результатам ряда международных мультицентровых рандомизированных исследований сформулированы показания к проведению реконструктивных операций на БЦА и тактика ведения пациентов в послеоперационном периоде. Ряд пациентов с атеросклеротическим поражением БЦА имеют сниженные когнитивные функции. Целью работы явилось сравнение когнитивных функций у пациентов, у которых использован различный хирургический подход при лечении стенозирующего атеросклероза ВСА.МАТЕРИАЛ И МЕТОДЫ. Изучены высшие психические функции у 116 больных со стенозирующим односторонним или двухсторонним поражением ВСА. Исследование КФ проводилось до каротидной эндартерэктомии (КЭЭ, группа 1, n=73) и транслюминальной баллонной ангиопластики ВСА (ТБА ВСА, группа 2, n=43), а также на 5–7-е сут и 30–31-е сут после реваскуляризации головного мозга. Для оценки общей тяжести когнитивных нарушений использовались суммарные показатели основных скрининговых нейропсихологических тестов: Mini Mental State Examination — MMSE; Montreal Cognitive Assasment — МоСА; Батарея тестов для оценки лобной дисфункции (БТЛД); шкала (тест-опросник) депрессии Бека и шкала Гамильтона.РЕЗУЛЬТАТЫ. Нарушения при нейропсихологическом исследовании до операции обнаружены у 98% больных. Выявлено исходно сопоставимое состояние высших психических функций в группах с КЭЭ и ТБА ВСА.При MMSE2 выявлено статистически значимое улучшение показателей в группе 1 как в сравнении с исходными данными (p<0,05), так и в сравнении с показателями второго тестирования груп пы 2. При этом сравнение данных MMSE1 и MMSE2 в группе 2 статистически значимых отличий не выявило (p>0,05). Данные по MMSE3 показали наличие статистически значимого улучшения в результатах тестирования в каждой из групп в сравнении с данными MMSE1 и MMSE2. При этом показатели MMSE3 были статистически значимо лучше (p<0,01) в группе 1, чем в группе 2.При MoCA-тестировании пациентов наибольшие отличия в результатах тестирования получены в группе 1, где имело место статистически значимое снижение когнитивной дисфункции в тестах MoCA1, MoCA2, MoCA3 и внутри группы (p<0,05), так и в сравнении данных MoCA3 между группой КЭЭ и группой ТБА (p><0,01). Тесты на оценку лобной дисфункции (БТЛД-тестирование) показали, что тест БТЛД2 составил 16,2 балла в группе с КЭЭ и 14,6 балла в группе с ТБА ВСА. Значительное улучшение показателей мнестических функций отмечалось при осмотре пациентов в динамике БТЛД3: 17,3 балла и 15,6 балла соответственно. Различия в обоих случаях оказались статистически значимыми (p<0,05).При оценке по шкалам Гамильтона и Бека в обеих группах пациентов отмечался умеренный уровень тревоги и депрессивной подавленности (по шкале Гамильтона 12,4 и 15,8 балла (p><0,05), по шкале Бека 12,3 и 14,4 балла (p>0,05)). При втором тестировании отмечалось статистически значимое снижение уровня депрессивного настроя пациентов (по шкале Гамильтона 8,4 и 13,8 балла (p<0,05), по шкале Бека 9,1 и 11,2 балла (p<0,05) статистически значимо в обоих случаях). При третьем тестировании пациентов в обеих группах отмечалось улучшение психологического фона. В группе с КЭЭ по шкале Гамильтона уровень достигал 5,8 балла, что в сравнении с группой с ТБА ВСА (11,0 баллов) имело наиболее статистически значимую разницу (p<0,001). Показатели, полученные при тестировании по шкале Бека, имели статистически значимые отличия (p<0,05) как внутри групп в динамике, так и между группами 1 и 2.ЗАКЛЮЧЕНИЕ. При сравнении ВПФ у пациентов, которым был использован различный хирургический подход (КЭЭ и трансбалонная ангиопластика) при лечении стенозирующего атеросклеротического поражения ВСА нами было выявлено, что: максимальное улучшение высших психических функций формируется к 30-м сут послеоперационного периода по сравнению с дооперационными по- казателями; наиболее значимое улучшение показателей тестов высших психических функций к 30-м сут послеоперационного периода отмечается в группе, в которой в качестве метода хирургической реваскуляризации головного мозга была использована КЭЭ

Alignathon: A competitive assessment of whole-genome alignment methods

© 2014 Earl et al. Multiple sequence alignments (MSAs) are a prerequisite for a wide variety of evolutionary analyses. Published assessments and benchmark data sets for protein and, to a lesser extent, global nucleotide MSAs are available, but less effort has been made to establish benchmarks in the more general problem of whole-genome alignment (WGA). Using the same model as the successful Assemblathon competitions, we organized a competitive evaluation in which teams submitted their alignments and then assessments were performed collectively after all the submissions were received. Three data sets were used: Two were simulated and based on primate and mammalian phylogenies, and one was comprised of 20 real fly genomes. In total, 35 submissions were assessed, submitted by 10 teams using 12 different alignment pipelines. We found agreement between independent simulation-based and statistical assessments, indicating that there are substantial accuracy differences between contemporary alignment tools. We saw considerable differences in the alignment quality of differently annotated regions and found that few tools aligned the duplications analyzed. We found that many tools worked well at shorter evolutionary distances, but fewer performed competitively at longer distances. We provide all data sets, submissions, and assessment programs for further study and provide, as a resource for future benchmarking, a convenient repository of code and data for reproducing the simulation assessments

Personalized peptide-based vaccination for treatment of colorectal cancer: rational and progress

Colorectal cancer (CRC) is one of the most common cancers globally and is associated with a high rate of morbidity and mortality. A large proportion of patients with early stage CRC who undergo conventional treatments develop local recurrence or distant metastasis and in this group of advanced disease, the survival rate is low. Furthermore there is often a poor response and/or toxicity associated with chemotherapy and chemo-resistance may limit continuing conventional treatment alone. Choosing novel and targeted therapeutic approaches based on clinicopathological and molecular features of tumors in combination with conventional therapeutic approach could be used to eradicate residual micrometastasis and therefore improve patient prognosis and also be used preventively. Peptide-based vaccination therapy is one class of cancer treatment that could be used to induce tumor-specific immune responses, through the recognition of specific antigen-derived peptides in tumor cells, and this has emerged as a promising anti-cancer therapeutic strategy. The aim of this review was to summarize the main findings of recent studies in exciting field of peptide-based vaccination therapy in CRC patients as a novel therapeutic approach in treatment of CRC

Analysis of canonical and non-canonical splice sites in mammalian genomes

A set of 43 337 splice junction pairs was extracted from mammalian GenBank annotated genes. Expressed sequence tag (EST) sequences support 22 489 of them. Of these, 98.71% contain canonical dinucleotides: GT and AG for donor and acceptor sites, respectively; 0.56% hold non-canonical GC-AG splice site pairs; and the remaining 0.73% occurs in a lot of small groups (with a maximum size of 0.05%). Studying these groups we observe that many of them contain splicing dinucleotides shifted from the annotated splice junction by one position. After close examination of such cases we present a new classification consisting of only eight observed types of splice site pairs (out of 256 a priori possible combinations). EST alignments allow us to verify the exonic part of the splice sites, but many non-canonical cases may be due to intron sequencing errors. This idea is given substantial support when we compare the sequences of human genes having non-canonical splice sites deposited in GenBank by high throughput genome sequencing projects (HTG). A high proportion (156 out of 171) of the human non-canonical and EST-supported splice-site sequences had a clear match in the human HTG. They can be classified after corrections as: 79 GC-AG pairs (of which one was an error that corrected to GC-AG), 61 errors that were corrected to GT-BG;canonical pairs, six AT-AC pairs (of which two were-errors that corrected to AT-AC), one case was produced from non-existent intron, seven cases were found in HTG that were deposited to GenBank and finally there were only two cases left of supported non-canonical splice sites. If we assume that approximately the same situation is true for the whole: set of annotated mammalian non-canonical splice-sites, then the 99.24% of splice site pairs should be GT-AG, 0.69% GC-AG, 0.05% AT-AC and finally only 0.02% could consist of other types of non-canonical splice sites. We analyze several characteristics of EST-verified splice sites and build weight matrices for the major groups, which can be incorporated into gene prediction programs. We also present a set of EST-verified canonical splice sites larger by two orders of magnitude than the current one (22 199 entries versus similar to 600) and finally, a set of 290 EST-supported non-canonical splice sites, Both sets should be significant for future investigations of the splicing mechanism

CLINICAL AND IMMUNOLOGICAL ASPECTS OF T C ELL-BASED VACCINE THERAPY IN PATIENTS WITH PROGREDIENT MULTIPLE SCLEROSIS

Abstract. Thirty-nine patients with a progredient clinical form of chronic multiple sclerosis (MS) were subject to multiple immunization with autologous polyclonal T-cell vaccines. Two years after initiating the vaccine therapy, no evidence for disease progression was noted in 16 patients (41% of total). Neurological improvement was observed in five cases (13%) from the vaccine-treated group. Of 22 control MS patients who did not receive the immunotherapy, only 6 persons (27%) exhibited stabilization of their clinical state. Clinical improvement was not detectable among this group of MS patients. A group of twenty-six MS patients was treated with Rebif, without evidence of disease progression in eleven cases (42%), and distinct neurological improvement noted in one patient (4%) from this group. One year after starting the vaccine therapy, a rise in serum IL-10 was detected in vaccine-treated patients, whereas IL-17 and IL-18 serum levels remained within the initial ranges. A correlation between the serum levels of anti-myelin antibodies and appropriate anti-idiotypic antibodies was revealed in these patients. In general, the results obtained suggest polyclonal T-cell vaccination as a potentially effective treatment approach, both at early and more advanced stages of the disorder