EFFECTIVENESS OF DIABETIC KETOACIDOSIS TREATMENT WITH LOW-DOSIS VENOUS INSULIN INFUSIONS

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Morphometric characterization of fibrinogen's αc regions and their role in fibrin self-assembly and molecular organization

© 2017 The Royal Society of Chemistry. The flexible C-terminal parts of fibrinogen's Aα chains named the αC regions have been shown to play a role in fibrin self-assembly, although many aspects of their structure and functions remain unknown. To examine the involvement of the αC regions in the early stages of fibrin formation, we used high-resolution atomic force microscopy to image fibrinogen and oligomeric fibrin. Plasma-purified full-length human fibrinogen or des-αC fibrinogen lacking most of the αC regions, untreated or treated with thrombin, was imaged. Up to 80% of the potentially existing αC regions were visualized and quantified; they were highly heterogeneous in their length and configurations. Conversion of fibrinogen to fibrin was accompanied by an increase in the incidence and length of the αC regions as well as transitions from more compact conformations, such as a globule on a string, to extended and more flexible offshoots. Concurrent dynamic turbidimetry, confocal microscopy, and scanning electron microscopy revealed that trimming of the αC regions slowed down fibrin formation, which correlated with longer protofibrils, thinner fibers, and a denser network. No structural distinctions, except for the incidence of the αC regions, were revealed in the laterally aggregated protofibrils made of the full-length or des-αC fibrinogens, suggesting a pure kinetic effect of the αC regions on the fibrin architecture. This work provides a structural molecular basis for the promoting role of the αC regions in the early stages of fibrin self-assembly and reveals this stage of fibrin formation as a potential therapeutic target to modulate the structure and mechanical properties of blood clots

Candidate vaccine construction against tick-born encephalitis based on hybrid recombinant flagG-protE-protein

The present work describes the construction of the gene encoding the recombinant protein flagG­protE, its synthesis, purification and study. The recombinant flagG­protE protein is a promising molecule for developing a candidate recombinant vaccine against tickborne encephalitis by the ability to bind to monoclonal antibodies (MCA) against native protein E of tick­borne encephalitis virus. The antigenic determinants of two recombinant proteins were studied: protE and flagG­protE using a panel of 8 MCA. The recombinant protein protE comprises the tick­borne encephalitis virus envelope protein and the flagGprotE recombinant protein has an additional flagG domain encoding flagellin G of Salmonella typhi. It was found that the MCA tested revealed epitopes on the recombinant protein protE. This indicates that the investigated recombinant protein has an antigenic structure similar to the antigenic structure of the native tick­borne encephalitis virus protein E. In the study of the recombinant protein flagG­protE by the ability to bind a panel of 8 MCA, only five of them react with epitopes of the tested protein. MCA 4F6, 7F10, and 6B9 did not recognize the corresponding epitope in the recombinant flagG­protE protein, while in the recombinant protein protE, these epitopes were detected successfully. Our data indicate that the antigenic structure of recombinant protE­protein can be changed under the influence of the flagellin domain, which in turn can lead to the unavailability of some antigenic determinants. This fact must be taken into account when constructing recombinant molecules with antigenic properties. Nevertheless, the fundamentally important regions in the region of the fusion peptide and III domain are antigenically present on the surface of the recombinant protein. This should ensure the formation of neutralizing antibodies, and the presence of a complete amino acid sequence of protein E in the recombinant protein induces the formation of a T­cell immune response. The emergence of a new generation of vaccines against tick­borne encephalitis with a higher level of safety and immunogenicity will improve the vaccine prevention of the population from tick­borne encephalitis

Analysis of the efficiency of IFV techniques (ICSI, PICSI) in the assisted reproductive technology programs

The male factor is the cause of infertility in 40 % of cases. This study is dedicated to the efficiency assessment of ICSI and PICSI techniques and the determination of indications of these methods. There are some relative indicationsfor ICSI in clinical practice: advanced reproductive age of patients, a small number of obtained oocytes, prolonged infertility, repeated attempts of in vitro fertilization. However, normal sperm values are registered in 71.3 % of cases. PICSI method is more preferable at high level of DNA fragmentation and associated changes in sperm indicators. Nevertheless, the pregnancy rate after these methods was comparable - 19.3 % and 19.7 % respectively. The results indicate that the ICSI and PICSI methods do not have a negative impact on the quality of the embryos and do not increase the pregnancy rate

p53 Amino-Terminus Region (1–125) Stabilizes and Restores Heat Denatured p53 Wild Phenotype

BACKGROUND:The intrinsically disordered N-ter domain (NTD) of p53 encompasses approximately hundred amino acids that contain a transactivation domain (1-73) and a proline-rich domain (64-92) and is responsible for transactivation function and apoptosis. It also possesses an auto-inhibitory function as its removal results in remarkable reduction in dissociation of p53 from DNA. PRINCIPAL FINDINGS/METHODOLOGY:In this report, we have discovered that p53-NTD spanning amino acid residues 1-125 (NTD125) interacted with WT p53 and stabilized its wild type conformation under physiological and elevated temperatures, both in vitro and in cellular systems. NTD125 prevented irreversible thermal aggregation of heat denatured p53, enhanced p21-5'-DBS binding and further restored DBS binding activity of heat-denatured p53, in vitro, in a dose-dependent manner. In vivo ELISA and immunoprecipitation analysis of NTD125-transfected cells revealed that NTD125 shifted equilibrium from p53 mutant to wild type under heat stress conditions. Further, NTD125 initiated nuclear translocation of cytoplasmic p53 in transcriptionally active state in order to activate p53 downstream genes such as p21, Bax, PUMA, Noxa and SUMO. CONCLUSION/SIGNIFICANCE:Here, we showed that a novel chaperone-like activity resides in p53-N-ter region. This study might have significance in understanding the role of p53-NTD in p53 stabilization, conformational activation and apoptosis under heat-stress conditions

Nuclear and Chloroplast Microsatellites Show Multiple Introductions in the Worldwide Invasion History of Common Ragweed, Ambrosia artemisiifolia

BACKGROUND: Ambrosia artemisiifolia is a North American native that has become one of the most problematic invasive plants in Europe and Asia. We studied its worldwide population genetic structure, using both nuclear and chloroplast microsatellite markers and an unprecedented large population sampling. Our goals were (i) to identify the sources of the invasive populations; (ii) to assess whether all invasive populations were founded by multiple introductions, as previously found in France; (iii) to examine how the introductions have affected the amount and structure of genetic variation in Europe; (iv) to document how the colonization of Europe proceeded; (v) to check whether populations exhibit significant heterozygote deficiencies, as previously observed. PRINCIPAL FINDINGS: We found evidence for multiple introductions of A. artemisiifolia, within regions but also within populations in most parts of its invasive range, leading to high levels of diversity. In Europe, introductions probably stem from two different regions of the native area: populations established in Central Europe appear to have originated from eastern North America, and Eastern European populations from more western North America. This may result from differential commercial exchanges between these geographic regions. Our results indicate that the expansion in Europe mostly occurred through long-distance dispersal, explaining the absence of isolation by distance and the weak influence of geography on the genetic structure in this area in contrast to the native range. Last, we detected significant heterozygote deficiencies in most populations. This may be explained by partial selfing, biparental inbreeding and/or a Wahlund effect and further investigation is warranted. CONCLUSIONS: This insight into the sources and pathways of common ragweed expansion may help to better understand its invasion success and provides baseline data for future studies on the evolutionary processes involved during range expansion in novel environments

ВЛИЯНИЕ РЕЗИСТИНА НА ТЕЧЕНИЕ ИШЕМИЧЕСКОЙ БОЛЕЗНИ СЕРДЦА У ПАЦИЕНТОВ С САХАРНЫМ ДИАБЕТОМ 2-го ТИПА

Relevance. Through dermal coronary intravascular revascularization by means of stenting is an effective method of treatment CHD patients with the type 2 diabets. At the same time frequency of stents restenosing for this special cohort of patients is fluctuating from 12 by 40 %.Objective. To study prognostic significance of the new biomarker of intravascular inflammation of resisin in blood of CHD patients with DM 2 who suffered from stenting. Material and methods. In the study 60 patients (48 men and 12 women, in the middle age 60.9 years) with CHD and DM2 are included. The patients were divided into two groups: Patients with positive progress of comorbide pathology belonged to the first group (n = 30); in the second group (n =3 0) patients with unfavorable progress of cardiac vessel pathology were included. The further observation was carried during 12 months. Content of all the patients in blood of resistin with help of enzyme immunoassay analysis was determined, as well as lipid blood serum mixture and additional metabolic risk factors. By the indications control a coronary angiography was conducted. Restenosis of coronary arteries was counted as the narrowing of a coronal artery lumen of ≥70% in the place of an intervention.The result. Unfavorable flow, including restenosis of coronary arteries was revealed in 30 cases (50%). ROC-analysis showed great predictive significance of resistin – (area under a curve >5/35, Sensitivity 86.2, Specificit 70.0) in development of constrictive coronary atherosclerosis restenosis of coronary arteries after their stenosis.The conclusion. The study of resistin level in DM patients blood for the valuation of the disease prognosis and optimization of the tactics for choice of coronary pathology treatment seems expedient and informative. Чрескожная эндоваскулярная коронарная реваскуляризация посредством стентирования является эффективным методом лечения ишемической болезни сердца (ИБС) у больных сахарным диабетом 2-го типа (СД-2). Вместе с тем, частота рестенозирования стентов в этой особой когорте пациентов колеблется от 12 до 40%. Цель исследования – изучить прогностическую значимость нового биомаркера внутрисосудистого воспаления резистина в крови больных ИБС с СД-2, перенесших стентирование.Материал и методы. В исследование включены 60 больных с ИБС, ассоциированной с СД-2.  Проспективное наблюдение проводилось в течение 12 мес. У всех пациентов определялось содержание в крови резистина с помощью иммуноферментного анализа, а также оценивался липидный состав сыворотки крови и дополнительные метаболические факторы риска. Всем обследованным выполнено стентирование коронарных артерий. Рестенозом коронарных артерий считали сужение просвета венечной артерии более 70% в месте вмешательства. По результатам обследования пациенты были распределены на две группы: в 1-ю группу вошли 30 пациентов с благоприятным течением патологии; во 2-ю включены 30 больных с неблагоприятным течением сердечно-сосудистой патологии.Результаты. Анализ патогенетической значимости резистина в качестве биологического маркера рестеноза стентов у больных СД-2 показал, что у пациентов 1-й группы, у подавляющего большинства которых отсутствовала гиперрезистинемия, не регистрировались нестабильная стенокардия, повторные инфаркты миокарда, прогрессирование хронической сердечной недостаточности. Во 2-й группе нестабильная стенокардия регистрировалась у 24 (80%) пациентов, повторный нефатальный ИМ, мозговой инсульт не зарегистрированы, летальный исход имел место в 1 случае (3%), рестеноз стента – в 6 (20%), прогрессирование атеросклероза – в 16 случаях (53,3%). Таким образом, явной гиперрезистенимии, диагностировавшейся во 2-й группе у больных ИБС с СД-2, сопутствовало неблагоприятное течение патологии по сравнению с 1-й группой.Заключение. Проведенное исследование показало, что оценка уровня в крови резистина в качестве независимого маркера рестеноза коронарных стентов в дополнение к объективным клиническим критериям обсуждаемой патологии позволяет стратифицировать степень риска неблагоприятного течения ИБС у пациентов, перенесших коронарное стентирование и выделить особые когорты больных для целевого, более интенсивного наблюдения и осуществления обоснованного патогенетического лечения

New approaches in the diagnosis and treatment of latent tuberculosis infection

With nearly 9 million new active disease cases and 2 million deaths occurring worldwide every year, tuberculosis continues to remain a major public health problem. Exposure to Mycobacterium tuberculosis leads to active disease in only ~10% people. An effective immune response in remaining individuals stops M. tuberculosis multiplication. However, the pathogen is completely eradicated in ~10% people while others only succeed in containment of infection as some bacilli escape killing and remain in non-replicating (dormant) state (latent tuberculosis infection) in old lesions. The dormant bacilli can resuscitate and cause active disease if a disruption of immune response occurs. Nearly one-third of world population is latently infected with M. tuberculosis and 5%-10% of infected individuals will develop active disease during their life time. However, the risk of developing active disease is greatly increased (5%-15% every year and ~50% over lifetime) by human immunodeficiency virus-coinfection. While active transmission is a significant contributor of active disease cases in high tuberculosis burden countries, most active disease cases in low tuberculosis incidence countries arise from this pool of latently infected individuals. A positive tuberculin skin test or a more recent and specific interferon-gamma release assay in a person without overt signs of active disease indicates latent tuberculosis infection. Two commercial interferon-gamma release assays, QFT-G-IT and T-SPOT.TB have been developed. The standard treatment for latent tuberculosis infection is daily therapy with isoniazid for nine months. Other options include therapy with rifampicin for 4 months or isoniazid + rifampicin for 3 months or rifampicin + pyrazinamide for 2 months or isoniazid + rifapentine for 3 months. Identification of latently infected individuals and their treatment has lowered tuberculosis incidence in rich, advanced countries. Similar approaches also hold great promise for other countries with low-intermediate rates of tuberculosis incidence