Single-point substitution F97M leads to in cellulo crystallization of the biphotochromic protein moxSAASoti.

To enhance the photoconversion performance of biphotochromic moxSAASoti protein, a substitution F97 M was introduced. In addition to enhancing the target properties, this substitution also resulted in the crystallization of the recombinant protein within living HeLa cells (also referred to as in cellulo crystallization). The phenomenon of protein crystallization in living cells is not unique, yet the mechanisms and application of in cellulo crystallization remain significant for further research. However, in cellulo crystallization is atypical for fluorescent proteins and detrimental for their biotechnological application. The objective of this study was to elucidate the underlying mechanisms responsible for the crystallization of moxSAASotiF97Min cellulo. For this purpose, the crystal structure of the green form of biphotochromic protein moxSAASotiF97M was determined at high resolution, which surprisingly has a space group, different from those of parent mSAASotiC21N. The analysis provided allowed to propose a mechanism of new crystal contacts formation, which might be a cause of in cellulo protein crystallization.

Additional booster doses in patients with chronic lymphocytic leukemia induce humoral and cellular immune responses to SARS-CoV-2 similar to natural infection regardless ongoing treatments: A study by ERIC, the European Research Initiative on CLL.

Profound immune dysregulation and impaired response to the SARS-CoV-2 vaccine put patients with chronic lymphocytic leukemia (CLL) at risk of severe COVID-19. We compared humoral memory and T-cell responses after booster dose vaccination or breakthrough infection. (Green) Quantitative determination of anti-Spike specific antibodies. Booster doses increased seroconversion rate and antibody titers in all patient categories, ultimately generating humoral responses similar to those observed in the postinfection cohort. In detail, humoral response with overscale median antibody titers arose in >80% of patients in watch and wait, off-therapy in remission, or under treatment with venetoclax single-agent. Anti-CD20 antibodies and active treatment with BTK inhibitors (BTKi) represent limiting factors of humoral response, still memory mounted in ~40% of cases following booster doses or infection. (Blue) Evaluation of SARS-CoV-2-specific T-cell responses. Number of T-cell functional activation markers documented in each patient. The vast majority of patients, including those seronegative, developed T-cell responses, qualitatively similar between treatment groups or between vaccination alone and infection cases. These data highlight the efficacy of booster doses in eliciting T-cell immunity independently of treatment status and support the use of additional vaccination boosters to stimulate humoral immunity in patients on active CLL-directed treatments.

The Relationship of p62 Gene Expression with Integrity of Mitochondrial DNA and the Level of Lipid Peroxidation Products in Skeletal Muscles of Rats of Different Ages Exposed to Different Feeding Protocols.

Sequestosome-1 (SQSTM1/p62) is one of the most important multifunctional proteins, which is necessary to maintain mitochondrial stability by eliminating damaged mitochondria through mitophagy. We studied the influence of age and diet on the expression of the p62 gene in the femoral and abdominal muscles of rats, as well as the integrity of some mitochondrial components. In the femoral muscles of 24-month-old rats receiving restricted ration, the expression of the p62 gene increased. We assume that activation of mitophagy contributed to a decrease in the levels of oxidative damage to mitochondrial DNA and LPO intensity in the femoral muscles of 24-month-old rats.

[Myocardial revascularization in chronic coronary artery disease. State of art].

The review addresses debatable issues of myocardial revascularization in chronic forms of ischemic heart disease, shows major differences between percutaneous coronary intervention and coronary artery bypass grafting in terms of long-term prognosis, and the dependence of the results on the clinical profile of the disease. The review of current publications demonstrates advantages of open surgery in long-term survival and prevention of adverse outcomes in target groups of patients.

[The teacher (scientific forensic legacy of professor M.I. Avdeev)]. / Ob uchitele (nauchnoe sudebno-meditsinskoe nasledie professora M.I. Avdeeva).

Main directions of scientific and scientific-practical activity of the professor Mikhail Ivanovich Avdeev are described in the article. These include: a set of scientific tasks to justify the staff and organizational structure, organization and justification of the expert work content in the specialized military forensic service, including the training programs development for specialization and thematic improvement of forensic experts; determining the limits of competence for forensic experts in determining the violent death type; systematization of causes and death conditions; establishing structure of sudden death causes in young persons; assessment of the pathogenetic role of trauma and pathology in the development of basal subarachnoid hemorrhages; formulation of the conceptual base content for forensic medicine; justification of a scientifically based sequence of forensic medicine reproduction; creation of a scientific school of military forensic experts; preparation and publication of about 50 textbooks, manuals and, monographs on forensic medicine, including the fundamental work «Forensic Medicine Course¼, «Forensic Examination of Living People¼, «Forensic Corpse Examination¼.

[Injuries after the biological exposure]. / Povrezhdeniya ot biologicheskogo vozdeistviya.

The aim of the scientific work is to justify the need to highlight the topic of injuries after the biological exposure in forensic medicine. The definition of the «biological trauma¼ concept is presented - it means impairment of the body structure and function as a result of a special injury effects typical for wildlife representatives (animals, plants). Biological exposure includes antigenic, toxin, allergic, bioelectric, bioorganic exposures and their combinations. Biological injuries should be distinguished from mechanical injuries caused by small, medium, and large mammals and reptiles. Options of the antemortem and postmortem biological factor effects are considered. The qualitative limits of the postmortem period are defined. A new method of forensic reconstruction of postmortem conditions is proposed. Forensic entomological, forensic microbiological examinations, and their complex combination with forensic examination are isolated as independent methods.

[Terminological and conceptual ambiguity of criminally-remedial regulation of the expert conclusion contents]. / Terminologicheskaya i smyslovaya neopredelennost' ugolovno-protsessual'noi reglamentatsii soderzhaniya zaklyucheniya eksperta.

Criminally-remedial law requirements to the expert conclusion are analyzed. A number of terms contained in the law that do not have an unambiguous interpretation are highlighted including the content of the expert conclusion, result, outcomes, methodology, method. The author definitions of these concepts are proposed.

[State and several prospects of studying pediatric pathology in forensic pediatrics]. / Sostoyanie i nekotorye perspektivy izucheniya patologii detskogo vozrasta v sudebnoi pediatrii.

Studies of Russian and foreign literature have shown that the features of mechanical injuries and the course of diseases in different pediatric age categories remain insufficiently investigated in forensic pediatrics, taking into account their physiological characteristics and concomitant pathology. Based on this, it is proposed to develop the diagnostic criteria for both mechanical injuries and diseases of children as priority and promising areas in scientific research, taking into account the age-related physiological characteristics, as well as the effects of external factors on the course of the pathomorphological process; to create a single algorithm for a comprehensive pathomorphological analysis of pediatric pathology, which allows forensic experts to fully answer questions concerning the prescription of injuries, the pathogenesis of diseases, and determining the severity of the harm to the child's health.

[Pathomorphological characteristics of sequelae of the black mamba bite]. / Patomorfologicheskaya kharakteristika posledstvii ukusa chernoi mamby.

Pathomorphological changes in internal organs due to the toxic effects of the black mamba venom are nonspecific in nature and presume its neurotoxic and cardiotoxic effects with the development of the DIC syndrome, pulmonary edema, and brain edema in the terminal period. In forensic medicine, the development of specific diagnostic criteria, as well as an algorithm for detecting toxins of the black mamba venom in biological fluids and tissues of internal organs can become a promising topic for scientific research.

[On the prospects of the impedance monitoring method for determining the prescription of death coming]. / O perspektivnosti metoda impedansometrii dlya opredeleniya davnosti nastupleniya smerti.

The aim of the scientific work is to analyze the prospects of using the impedance monitoring method to determine the prescription of death coming. The conducted exploratory analysis allows hypothesizing the dependence of impedance values and dispersion factors for the diagnostic zones studies on the postmortem interval; it also suggests that it is possible to determine this interval for the objects (pig corpses) studies with a combination of impedance values and the corresponding factors. Among large mammals, a swine is the closest to a human from the point of analyzing the postmortem period process, and it can be recommended as an adequate human corpse model. When determining the correlation between the postmortem interval and the impedance parameters, it is the simplicity and reproducibility of the method, the absence of expensive equipment, its portability and the rapid receipt of results that allow this method to be applied at the scene, reinforcing traditional forensic methods for determining the prescription of death coming. Interpretation of impedance monitoring results can be used to analyze the biological nature of the postmortem period processes.

[Practical value of microorganisms for forensic purposes (on the example of microbial flora of bony remnants from the historic burial site)]. / Prakticheskoe znachenie mikroorganizmov dlya tselei sudebnoi meditsiny (na primere mikroflory kostnykh ostankov istoricheskogo zakhoroneniya).

Phenotypic signs of dominants isolated from the surface of bony remnants from the historic burial site were analyzed in order to expand data on the biodiversity of microorganisms in the microbial flora of bony remnants and to assess the possibility of using the results of microbiological analysis in the evidence base of forensic examination and forensic archaeology. It was detected that only Deuteromycota and Eubacteria colonized all types of surfaces in the samples of bone fragments from the historic burial site (with the age in the range of 90-95 years); with the abundance of micromycetes, the proportion of Eubacteria naturally decreased, while with the increased bacterial background counts the rate of micromycetes detection decreased. The insignificant amount of nutrients in the bony remnants led to the decrease in the number and biological diversity of microorganisms contaminating them; species adapted to a hard-to-reach organic substrate dominated there. During the process of bony remnants decomposition, when the conditions of their location changed, inter-species competition and specific recolonization occurred by species of microorganisms most adapted to a hard-to-reach organic substrate in the abiotic and biotic conditions of existence given. The results obtained are important for the descriptive ecology and biology of specific groups of microorganisms in the postmortem microbiome and form the basis for a more thorough study of complex communications between species of microorganisms in the necrobiome of bony remnants - in the future it will allow putting forward original hypotheses about the involvement of microbes in the circulation of matter and energy, as well as to apply the information obtained in the evidence base of forensic examination and forensic archaeology.

[On the issue of forensic evaluation of mandibular fractures complicated with infections]. / K voprosu sudebno-meditsinskoi otsenki iskhodov perelomov nizhnei chelyusti, oslozhnennykh infektsionnym protsessom.

The analysis of 387 cases of mandibular fractures complicated by posttraumatic infections is presented. These infections were quite diverse - from local suppuration of soft tissues surrounding the fracture to widespread processes affecting fatty tissue planes, including the anterior and posterior mediastinum. Infectious complications determine the injury outcome in each case - recovery, disability, or death. Two clinical observations with lethal outcomes are described.

[Main milestones of forensic ballistics history in Russia]. / Osnovnye vekhi istorii sudebno-meditsinskoi ballistiki v Rossii.

The purpose of the scientific work is to outline the main stages of the development of forensic examination of gunshot injuries in Russia. The analysis of the special literature from 1865 to the present time has demonstrated that the issue of forensic examination of gunshot injuries has been analyzed quite deeply by Russian forensic specialists. Meanwhile, expert practice sets forth tasks that are associated with the emergence of new firearm samples and new methods of laboratory and instrumental diagnosis.

[Dirofilariasis in forensic practice]. / Dirofilyarioz v sudebno-meditsinskoi praktike.

Parasitic diseases are rare in forensic practice. The article presents two observations of dirofilariasis in the facial area. Clinical signs, characteristics, surgical treatment, and description of the parasite extracted from the subcutaneous tissue of the paraorbital region are given.

[Rare cases of head traumas incited by propeller blades of small vessels]. / Redkie sluchai travm golovy, prichinennykh lopastyami grebnykh vintov malomernykh sudov.

2 cases of death in males due to multiple head impacts by propeller blades of small vessels are given. The set of the following signs provides a basis for confirming the above-mentioned mechanism and properties of the traumatic object: multiplicity, oblong shape, parallel step-like arrangement of injury lengths, acute-angled M-shaped or Y-shaped injury ends, uneven edges, sometimes with the formation of small flaps and abrasion, lumpy wound walls, multi-fragmented nature of fractures in a limited area of the skullcap and facial skull bones, the location of damage to intracranial structures in the projection of external injuries and fractures of the cerebral skull bones.

[The nutrition of population of Russia from public health position: international recommendation practice].

The Member States of the United Nations, while planning and implementing concrete measures to protect citizens health, nutrition included, experience need in sharing best practices and in coordinated recommendations, relevant to common actual challenges. The article analyzes and systematizes main official documents of the leading international organizations in the field of population nutrition from the standpoint of public health and health care in dynamics more than over 70 years. The particular emphasis is made on their implementation to adolescents and youths aged 10-24 years, who need risk approach application in resolving their urgent nutritional problems, in relationship with influencing environmental factors, lifestyle, bad habits, physical activity, etc. The policies, programs and other documents that are currently implementing in the area of numerous problems associated with malnutrition, promote adoption of their own commitments, which must be relevant, achievable, specific, time-bound, measurable. In this regard, adopted and implemented leading national documents in this area are analyzed. The practicability of implementation of the UN and WHO recommendations with their adaptation to multiple-aspect specifics of the country, as well as need in prolonging research in this area are demonstrated.

[On using the corpses of large mammals as model objects for studying the postmortem period]. / Ob ispol'zovanii trupov krupnykh mlekopitayushchikh v kachestve model'nykh ob"ektov pri izuchenii postmortal'nogo perioda.

THE AIM OF THE STUDY: Is to justify using the corpses of large mammals as model objects for studying the postmortem period. Similarities in processes occurring postmortem in human and swine corpses (decomposition stages and the structure of dominant necrophilic organisms inhabiting the corpse), as well as similar changes of relative impedance parameters for the cartilaginous tissue and musculoskeletal flap of swine and human corpses have been established. The results obtained allow recommending the swine corpse as an adequate human corpse model both for scientific studies and for solving specific scientific and practical issues arising in the practice of forensic examination when determining the prescription of death coming and the postmortem period conditions.

[Development of osteoplastic gene-activated 3D matrices based on adenoviral vectors]. / Razrabotka osteoplasticheskikh gen-aktivirovannykh 3D-matriksov na osnove adenovirusnykh vektorov.

Gene therapy is one of the most promising approaches in regenerative medicine for the restoration of extensive bone defects in dentistry and maxillofacial surgery. Matrices obtained using three-dimensional printing from bioresorbable polymers, impregnated with adenoviral constructs with genes for osteoinductive factors, can ensure safe and effective formation of bone tissue. OBJECTIVE: To study the properties of three-dimensional matrices based on polylactic-co-glycolic acid and adenoviral constructs with the GFP gene in vitro. MATERIALS AND METHODS: The matrices were obtained by antisolvent three-dimensional printing. Transduction efficiency was assessed by fluorescence microscopy and flow cytometry. The cytocompatibility of the matrices was assessed by the MTT test and by staining cells with fluorescent dyes. RESULTS: Matrices based on polylactic-co-glycolic acid have high cytocompatibility on adipose tissue-derived mesenchymal stem cells. Impregnation of adenoviral vectors with the green fluorescent protein gene in 3D matrices ensures the release of viral particles within a week, maintaining their high transducing ability. CONCLUSION: The developed method for obtaining gene-activated matrices can serve as the basis for the creation of effective osteoplastic materials for bone regeneration.

The role of Tyr102 residue in the functioning of bacterial NAD+-dependent formate dehydrogenase of Pseudomonas sp. 101.

Once you have missed the first button …, you'll never manage to button up Johann Wolfgang von Goethe Formate oxidation is a final step of methanol oxidation in methylotrophic prokaryotes and is important for detoxification of formate in other organisms. The structural mechanism of the formate dehydrogenase (FDH) of Pseudomonas sp. 101 has been studied for about 30 years. In the active center of FDH, the oxidation of formic acid into carbon dioxide in a NAD+-dependent way takes place. Residues that form the active center of that enzyme, as well as those that form the so-called substrate channel, are engaged in the catalytic cycle. Our study allowed to characterize a new residue, Tyr102, involved in the work of the enzyme. This residue is located in the outer neck of the substrate channel (at the beginning of the path of the substrate to the active center) and acts as a "button" which connects two enzyme domains into an active, "buttoned up" conformation. Our study of the kinetic parameters of mutant enzymes has shown that Tyr102Phe substitution leads to an approximately 80-fold increase of the Michaelis constant relative to the native enzyme, unlike Phe311Trp and Phe311Tyr substitution of neighboring residue Phe311. Our analysis of the Tyr102Phe mutant in the open conformation by X-ray crystallography has shown that its overall fold remains almost the same as that of the native enzyme. Molecular dynamics simulations of the ternary complexes of the native FDH enzyme and its Tyr102Phe mutant showed that Tyr102Phe substitution results in the loss of an interdomain hydrogen bond between the Tyr102 and Gln313 residues, which, in turn, destabilizes the closed conformation and affects the isolation of the FDH active site from water molecules. Our structural investigations have shown that Tyr102Phe replacement also leads to the destruction of interdomain contacts of Phe102 with Phe311, Pro312 residues, and decreases the stability of the Leu103-Val127 beta bridge. Phylogenetic analysis also confirmed the importance of the Tyr102 residue for enzymes from the FDH family, in which it is absolutely conserved.

Influence of Gold Nanoparticles on the Tolerance of Wheat to Low Temperature.

It was shown for the first time that the treatment of winter wheat (Triticum aestivum L.) seeds with gold nanoparticles (average diameter 15.3 nm; solution concentration 20 µg/mL) increases plant tolerance to low temperature. It was found that an increase in tolerance under the influence of nanoparticles is accompanied by a number of changes depending on temperature conditions. In optimal temperature conditions, gold nanoparticles stimulated plant growth and the activity of the photosynthetic apparatus, whereas in conditions of low-temperature hardening (2°C, 7 days) they inhibited growth but maintained photosynthetic activity, contributing to the accumulation of soluble sugars (cryoprotectants) in the leaves. It is concluded that gold nanoparticles can be considered as adaptogens that increase plant tolerance to low temperatures; however, their effectiveness in this role and the subtle mechanisms of action require further study.