Design and assembly of plant-based COVID-19 candidate vaccines: reсent development and future prospects.

An outbreak of a new variant of the coronavirus infection, known as COVID-19, occurred at the end of 2019 in China, in the city of Wuhan. It was caused by the SARS-CoV-2 virus. This variant of the virus is characterized by a high degree of variability and, as the current situation with its spread across different regions of the globe shows, it can lead to a progressive spread of infection among the human population and become the cause of a pandemic. The world scientific community is making tremendous efforts to develop means of protection,prevention and treatment of this disease based on modern advances in molecular biology, immunology and vaccinology. This review provides information on the current state of research in the field of vaccine development against COVID-19 with an emphasis on the role of plants in solving this complex problem. Although plants have long been used by mankind as sources of various medicinal substances, in a pandemic, plant expression systems become attractive as biofactories or bioreactors for the production of artificially created protein molecules that include protective antigens against viral infection. The design and creation of such artificial molecules underlies the development of recombinant subunit vaccines aimed at a rapid response against the spread of infections with a high degree of variability. The review presents the state of research covering a period of just over two years, i. e. since the emergence of the new outbreak of coronavirus infection. The authors tried to emphasize the importance of rapid response of research groups from various scientific fields towards the use of existing developments to create means of protection against various pathogens. With two plant expression systems - stable and transient - as examples, the development of work on the creation of recombinant subunit vaccines against COVID-19 in various laboratories and commercial companies is shown. The authors emphasize that plant expression systems have promise for the development of not only protective means under conditions of rapid response (subunit vaccines), but also therapeutic agents in the form of monoclonal antibodies against COVID-19 synthesized in plant cells.

[Clinical Findings in Two patients with DSD 46XY caused by new variant of the Desert Hedgehog Gene and review of the literature of the role of DHH signaling pathway in sex development].

Mutations in the gene DHH are an extremely rare cause of disorders of sex development 46,XY (DSD,46XY). The article describes the clinical cases of two unrelated patients with gonadal dysgenesis 46,XY with female phenotype. By using a next generation sequencing method, in both cases the same biallelic variant substitution c. 419T>G in the DHH gene was revealed. Taking into account the data on the role of DHH in the formation of the nervous system, the diagnosis of minifascicular polyneuropathy at the preclinical stage was confirmed in both cases. These cases demonstrate the value of using NGS, which allows simultaneous analysis of a wide range of candidate genes in DSD and the diagnosis of comorbidities before the development of the clinical picture. These are the first descriptions of patients with mutations in the DHH gene in the Russian population.

Peculiarities of the Inflammatory Process in the Reproductive Organs of C57Bl/6 Female Mice with Experimental Tuberculosis.

Intravenous infection of C57Bl/6 female mice with M. tuberculosis H37Rv led to involvement of the lungs and dissemination of the tuberculous infection to the abdominal and pelvic organs. M. tuberculosis were detected in the lungs and spleen in 14, 35, and 90 days and in the uterine horns in 90 days after infection. Morphological analysis of organs showed successive development of exudative necrotic tuberculosis of the lungs, acute and chronic nonspecific inflammation in the reproductive organs (vagina, uterus, and uterine horns). The inflammatory process in the reproductive organs was associated with the development of anaerobic dysbiosis, that was most pronounced in 35 days after infection. Antituberculous therapy was followed by reduction of M. tuberculosis count in the lungs and spleen in 60 and 90 days after infection, eliminatation of M. tuberculosis in the uterine horns, arrest of nonspecific inflammation in female reproductive organs, recovery of the balance between aerobic and anaerobic microflora, and development of candidiasis of the urogenital mucosa.

State of research in the field of the creation of plant vaccines for veterinary use.

Transgenic plants as an alternative of costly systems of recombinant immunogenic protein expression are the source for the production of cheap and highly efficient biotherapeuticals of new generation, including plant vaccines. In the present review, possibilities of plant system application for the production of recombinant proteins for veterinary use are considered, the history of the "edible vaccine" concept is briefly summarized, advantages and disadvantages of various plant systems for the expression of recombinant immunogenic proteins are discussed. The list of recombinant plant vaccines for veterinary use, which are at different stages of clinical trials, is presented.

[Bcl-2 family proteins and mechanisms of the acceleration and inhibition of tumor progression in vivo].

Earlier the authors demonstrated that the process of tumor progression in vivo may be inhibited or accelerated depending on the conditions of tumor growth (accelerated by tumor cell dissemination or delayed in locally growing tumors). It was also shown that tumor progression is inhibited in case of bcl-2 gene transduction in tumor cells. In this study, the research into mechanisms of the acceleration or inhibition of tumor progression and the role that Bcl-2 family proteins may play in these phenomena was continued. The results of the study demonstrated the following 1) immediate in vivo activation of endogenous proapoptotic Bax protein in disseminated tumor cells, not protected by Bcl-2 against apoptosis, and its correlation with accelerated tumor progression; 2) complete suppression of in vivo Bax activation in tumor cells protected by Bcl-expression, and inhibited tumor progression; 3) alternative character of Bcl-2 and Bax expression under the conditions of accelerated and inhibited tumor progression. Thus, the data presented support the hypothesis that the rates of tumor progression in vivo are regulated depending on the initial anti- and proapoptotic programs of tumor cells.

[Vulvovaginal candidiasis in girls (guidelines for a practitioner)].

There are many publications on the problems of the vaginal candidiasis treatment. Still, the actuality is in the frequency of the relapses and registration of the pediatric cases. Unfortunately, the majority of the pediatrists prescribe antibiotics but either forget to prescribe the simultaneous use of antifungal drugs, or prescribe nystatin, an antimycotic, that is known to be active only in the intestine lumen. As a result, candidiasis of the stomatopharynx or vagina is developed. Of no less importance is the fact of marketing many generics in Russia, which often prove to be false after their detailed investigation. The paper presents specific characteristics pertaining to the origin and development of vulvovaginal candidiasis in girls and indicates drugs with proved efficacy in normalization of the vagina microbiocenosis in girls.

[Construction of DNA-fragments' libraries with complete genomes of different variola strains].

Libraries of hybrid plasmids carrying DNA fragments of complete genomes of 8 variola virus strain from the Russian Collection belonging to 2 epidemical types and isolated in various geographic regions of the world were obtained. Genomic sequences of variola virus can be thus preserved for a long time in a biologically safe form and provide the research work on studying the genetic organization of this unique virus and on developing modern methods for rapid detection of variola virus and other orthopoxviruses.

[Specific features of normal vaginal microflora in girls of preschool age]. / Osobennosti normal'noi mikroflory vlagalishcha u devochek doshkol'nogo vozrasta.

The study of the vaginal microbial cenosis in 20 healthy girls aged 3-7 years did not confirm the notion on the dominating role of cocci (including epidermal staphylococci). The associations of 2-5 different microorganisms represented by more than 20 species in an amount of 4-6 Ig PFC/g of discharge were established. In the overwhelming majority of the examinees (84.2%) the microbial associations of the vagina were found to contain bifidobacteria. Gram positive cocci (staphylococci and streptococci) took the 2nd and 3rd places in the isolation rates and were detected in vaginal associations in 78.9% of the girls. Staphylococci were represented by 5 coagulase-negative staphylococcal species with S. simulans and S. epidermidis prevailing. Hemolytic streptococci variants alpha and beta were isolated in the proportion of 2:1. The latter belonged to serogroups C and F. No S. aureus, Lactobacillus sp., streptococci of groups A and B, yeast-like fungi were detected. Genital mycoplasms (M. hominis) could rarely be found in the vaginal discharge of the girls aged 3-7 years (5.3%). No resident and transitory components could be isolated from the normal vaginal microflora and no quantitative domination of any bacterial species (genus) was shown. The concentrations of all organisms in this association were moderate or low.

[Fundamentals of medical rehabilitation of patients with asymmetrical deformation of the maxilla complicated by cross occlusion]. / Printsipy meditsinskoi reabilitatsii bol'nykh s nesimmetrichnymi deformatsiiami verkhnei cheliusti, oslozhnennymi perekrestnym prikusom.

The problem of medical rehabilitation of patients with asymmetrical deformation of the facial skull, complicated by cross occlusion, is discussed. The authors suggest osteotomy of the maxillary complex with translocation of the osteotomied fragment in the sagittal, transverse, and vertical planes with osteotomy to a distance needed for elimination of deep cross occlusion. This method in combination with orthodontic treatment repairs the anatomic and functional abnormalities and brings about good esthetic results.

Analysis of the monkeypox virus genome.

Monkeypox virus (MPV) belongs to the orthopoxvirus genus of the family Poxviridae, is endemic in parts of Africa, and causes a human disease that resembles smallpox. The 196,858-bp MPV genome was analyzed with regard to structural features and open reading frames. Each end of the genome contains an identical but oppositely oriented 6379-bp terminal inverted repetition, which similar to that of other orthopoxviruses, includes a putative telomere resolution sequence and short tandem repeats. Computer-assisted analysis was used to identify 190 open reading frames containing >/=60 amino acid residues. Of these, four were present within the inverted terminal repetition. MPV contained the known essential orthopoxvirus genes but only a subset of the putative immunomodulatory and host range genes. Sequence comparisons confirmed the assignment of MPV as a distinct species of orthopoxvirus that is not a direct ancestor or a direct descendent of variola virus, the causative agent of smallpox.

Human monkeypox and smallpox viruses: genomic comparison.

Monkeypox virus (MPV) causes a human disease which resembles smallpox but with a lower person-to-person transmission rate. To determine the genetic relationship between the orthopoxviruses causing these two diseases, we sequenced the 197-kb genome of MPV isolated from a patient during a large human monkeypox outbreak in Zaire in 1996. The nucleotide sequence within the central region of the MPV genome, which encodes essential enzymes and structural proteins, was 96.3% identical with that of variola (smallpox) virus (VAR). In contrast, there were considerable differences between MPV and VAR in the regions encoding virulence and host-range factors near the ends of the genome. Our data indicate that MPV is not the direct ancestor of VAR and is unlikely to naturally acquire all properties of VAR.

Species-specific differences in the structure of orthopoxvirus complement-binding protein.

Vaccinia virus complement-binding protein (VCP) is secreted from the cells infected with the virus and controls the complement activation reactions. This protein contains four short consensus repeats (SCR), typical of the protein family of complement activation regulators. Organization of the VCP genes/proteins of orthopoxviruses-monkeypox (MPV), variola, cowpox and vaccinia viruses-and their cellular homologues (DAF and C4BP) were studied comparatively. For this purpose, VCP genes of three MPV strains were sequenced. VCP gene sequences of other human-pathogenic orthopoxvirus species and strains determined earlier were involved in the analysis. It has been demonstrated that a premature termination of the MPV VCP open reading frame results in a truncated protein sequence carrying a deletion of the C-terminal SCR4. This is an essential distinction of MPV from other orthopoxvirus species.

The fingerprints of the host innate immunity on the cells of primary virus-induced tumors.

As shown earlier, the cells transformed in vitro by various oncogenes, during subsequent in vivo growth were gradually replaced by descendant tumor cells, which co-expressed highly increased H(2)O(2)-catabolizing antioxidant activity (H(2)O(2)(CA)), and the ability to release PGE(2) (PGE(S)) in contact with natural killer cells; v-src was the only oncogene, which in vitro induced cells transformation characterised with the expression of [H(2)O(2)(CA)+PGE(S)] phenotype. Expression of [H(2)O(2)(CA)+PGE(S)] phenotype was providing tumor cells with significantly increased resistance to cytotoxic activities of macrophages and NK cells. However, the possible involvement of [H(2)O(2)(CA)+PGE(S)] phenotype in primary carcinogenesis remained obscure. Here, using three models of the primary virus-induced Syrian hamster tumors we demonstrated that Rous Sarcoma Virus-induced tumors arising after short latent period expressed [H(2)O(2)(CA) + PGE(S)] phenotype at appearance. During the latent periods of SV40- and SA7(C8)-induced tumors the cells expressing [H(2)O(2)(CA)+PGE(S)] phenotype gradually replaced the original [H(2)O(2)(CA)+PGE(S)]-phenotype-negative cell populations. The effectiveness of such selection correlated with the duration of in vivo tumor development. Thus it was shown, that selection of tumor cells expressing [H(2)O(2)(CA)+PGE(S)] phenotype is beginning and may be completed during the latent period of primary carcinogenesis. Taken together, data of this and preceding our studies on [H(2)O(2)(CA)+PGE(S)] phenotype demonstrate that in vivo the host innate immunity antitumor reactions are apparently responsible for the selection of rare tumor cell variants capable to defend themselves against CTA of Mph and NK.