Finding RB/Rpi-blb1/Rpi-sto1-like sequences in conventionally bred potato varieties

The main objectives in potato breeding are increasing yield abilities and improving resistance to numerous pathogens and pests. Among them, the late blight caused by the Phytophthora infestans oomycete is one of the most destructive potato diseases both in Russia and worldwide. Wild relatives of cultivated potato are traditionally used in breeding as the source of valuable R genes conferring resistance to pathogens. Of particular interest are Mexican wild species because Mexico is the centre of origin and diversity of P. infestans and at the same time, it is the centre of potato species diversity.  Mexican wild potato species S. bulbocastanum and S. stoloniferum are an important source of the R genes conferring broad-spec trum resistance against various isolates of P. infestans (Rpi-blb1,  Rpi-blb2, Rpi-sto1). Recently these genes have been transferred into cultivated potato gene pool using the cisgene  approach. At the same time there is a high probability of finding geno types with the Rpi-sto1 gene (functional homologues of  Rpi-blb1) among conventionally bred varieties because for about 40 years S. stoloniferum has been used in breeding as a source of the Rysto and Ry-fsto genes of the extreme resistance to the most important viral pathogen PVY. In this study 188 potato varieties bred in Russia and in near-abroad countries were screened for the presence of six gene-specific markers of the RB/Rpi-blb1 =  Rpi-sto1 and Rpi-blb2 genes conferring broad-spectrum resistance against P. infestans, and for the markers linked to the Rysto and Ry-fsto genes conferring extreme resistance to PVY. In addition, a marker for detecting male sterile mitochondrial DNA type gamma derived from S. stoloniferum was used. The genotypes selected through the molecular markers were divided into four groups: (A) 13 PVY resistant varieties carrying diagnostic markers of the Rysto, Ry-fsto genes and having sterile mt-type gamma; (B) four varieties possessing mt-type gamma and not having the markers of the R genes introgressed from S. stoloniferum; (C) eight genotypes carrying five gene-specific markers for the RB/Rpi-blb1/= Rpi-sto1; (D) the rest 166 (86.9 %) varieties not possessing any of the diagnostic markers associated with the S. stoloniferum genetic material. The sequences of the Rpi-sto1- and BLB1 F/R-amplicons were identical in all the genotypes of group ‘C’ and showed respective 99 % and 100 % similarity to the corresponding fragments of the Rpi-sto1 and Rpi-blb1 genes from the GenBank database. Among the genotypes of group ‘C’ various mt-types were detected, and some of them were male fertile

The Role of Vesicles in Transporting of Cholera Toxin

The review reports on the secretion pathways of the main virulence factor of Vibrio cholerae, cholera toxin, both through the two-stage Sec-dependent type 2 secretion system and with the help of vesicles of the outer membrane of V. cholerae. The ways of toxin transfer into the host organism, depending on its form, are discussed. The well-studied free soluble cholera toxin is secreted extracellularly and transmitted in a GM1-dependent manner through cholesterolrich lipid rafts. The transfer of cholera toxin associated with vesicles has advantages over free toxin, because substances inside the outer membrane vesicles are protected from external proteases and host antibodies by the membrane that forms the vesicle. Vesicular transporting of cholera toxin into the target cell occurs via clathrin-dependent, caveolin-dependent and lipid raft-dependent endocytosis. The specific transport route is determined by the structure of the vesicles. Clathrindependent endocytosis is described for V. cholerae strains cultivated at low osmolarity of the medium, whose outer membrane vesicles contain the cholera toxin subunit A inside. Lipid raft-dependent endocytosis is characteristic of vesicles in which cholera toxin is located on the surface. In addition, endocytosis of V. cholerae outer membrane vesicles through structures known as caveolae is presented

Comparative safety evaluation of intranasal and intramuscular immunisation with Ad26 and Ad5-vectored vaccines to prevent coronavirus infection

Scientific relevance. Intranasal vaccination may dramatically change current approaches to mass immunisation against various infections, shifting the focus from inducing systemic humoral and cellular immune responses to developing mucosal immunity and protective barriers on the mucous membranes, which are entry points for pathogens. Therefore, the safety of switching from intramuscular to intranasal vaccination should be evaluated.Aim. This study aimed to compare the safety of intranasal and intramuscular vaccination using the intermediate results of the phase III VCI-COV-III clinical trial in healthy volunteers.Materials and methods. The evaluation of the safety profile was based on the adverse events and adverse drug reactions (ADRs) documented by investigators in the interim report on the randomised, double-blind clinical trial of the Salnavac intranasal vaccine (InV) and the Gam-COVID-Vac intramuscular vaccine (ImV) against coronavirus infection. The report covered 42 days of observation in 137 healthy volunteers.Results. ADRs were reported for 17/68 (25%) and 30/69 (43.5%) volunteers in the InV group and the ImV group, respectively (p=0.036). The ADRs reported for the InV group were systemic and local, short-term, mostly mild, and generally consistent in incidence and nature with the ADRs typically observed with other nasal products.Conclusions. According to the preliminary data obtained in the clinical trial, the InV demonstrates a high level of safety. Its safety profile is comparable with those of other intranasal and intramuscular vaccines for the prevention of coronavirus infection

Выявление нетуберкулезных микобактерий, циркулирующих в разных регионах Сибири, и анализ их лекарственной устойчивости

The objective of the study: to identify the frequency of various species of non-tuberculous mycobacteria (NTM) isolated in the patients of TB services in Siberian Region during 2014-2018, to determine the correlation between rates of resistance to different antibacterial drugs.Subjects and methods. PCR hybridization on strip strips (Hain Lifescience GenoType Mycobacterium CM / AS) was used to genotype 475 NTM isolates of patients from various TB hospitals inSiberia. Drug susceptibility was tested by minimum inhibitory concentrations (MIC) on culture with serial micro-dilutions in a liquid medium using TREK Diagnostic Systems kits. 64 panels were used for slow-growing mycobacteria (SLOWMYCO), 46 - for fast-growing ones(RAPMYCO). Data processing was performed using the APL of 3.5.1 and the data analysis software of Rstudio, version 1.1.442.Results of the study1. Tomsk Region differs from the other considered regions ofSiberiadue to higher frequency of M. intracellulare among isolates identified by PCR hybridization. In other regions, M. fortuitum prevails among isolated.2. Incomparison with the reference data (CLSI M62, 2018), differences in the MIC of some NTM species isolated from patients in Siberia and theFar Eastwere observed, most likely related to the regional characteristics of these isolates.3. HTM isolates resistant to moxifloxacin are also resistant to linezolid, trimethoprim, and ciprofloxacin.4. Fast-growing clarithromycin resistant HTM isolates are predominantly sensitive to moxifloxacin, ciprofloxacin, and trimethoprim.Цель исследования: выявить частоту видов нетуберкулезных микобактерий (НТМ), выделенных от пациентов противотуберкулезных учреждений Сибирского региона за период 2014-2018 гг., определить корреляционные взаимосвязи между показателями резистентности к разным антибактериальным препаратам.Материалы и методы. Методом ПЦР-гибридизации на стрип-полосках (Hain Lifescience GenoType Mycobacterium CM/AS) проведено генотипирование 475 изолятов НТМ, выделенных от пациентов разных противотуберкулезных стационаров Сибири. Лекарственную устойчивость тестировали методом минимальных ингибирующих концентраций (МИК), культуральным методом серийных микроразведений в жидкой питательной среде с применением наборов TREK Diagnostic Systems. Использовали 64 панели для медленнорастущих микобактерий (SLOWMYCO), 46 – для быстрорастущих (RAPMYCO). Обработку данных проводили с использованием языка программирования 3.5.1 и среды анализа данных Rstudio, версии 1.1.442.Результаты исследования.1. Томская область отличается от остальных рассматриваемых регионов Сибири большей частотой встречаемости M. intracellulare среди изолятов, идентифицированных методом ПЦР-гибридизации. В остальных регионах наиболее часто встречающимся видом является M. fortuitum.2. В сравнении с референсными данными (CLSI M62, 2018) получены отличия МИК некоторых видов НТМ, выделенных у больных на территориях Сибири и Дальнего Востока, скорее всего связанные с региональными особенностями данных изолятов.3. Изоляты НТМ, устойчивые к моксифлоксацину, являются также резистентными к линезолиду, тримептоприму, ципрофлоксацину.4. Быстрорастущие изоляты НТМ, устойчивые к кларитромицину, преимущественно являются чувствительными к моксифлоксацину, ципрофлоксацину и триметоприму

Assembling the puzzle of superconducting elements: A Review

Superconductivity in the simple elements is of both technological relevance and fundamental scientific interest in the investigation of superconductivity phenomena. Recent advances in the instrumentation of physics under pressure have enabled the observation of superconductivity in many elements not previously known to superconduct, and at steadily increasing temperatures. This article offers a review of the state of the art in the superconductivity of elements, highlighting underlying correlations and general trends.Comment: Review, 10 pages, 11 figures, 97 references; to appear in Superc. Sci. Techno

Experience with olaparib in a patient with luminal HER2-positive metastatic breast cancer

Hereditary breast cancer (BC) accounts for about 5-10% of cases. BRCA-associated tumors have been identified as a separate group of malignant neoplasms with distinctive clinical manifestations and specific treatment features. Understanding of biological mechanisms leading to cancer in BRCA1/2 mutation carriers and discovery of potential molecular targets, such as poly (ADP-ribose) polymerase (PARP), involved in base excision repair mechanisms, led to the development of a new class of targeted drugs belonging to the PARP inhibitors group. PARP inhibition leads to the preservation of single-stranded DNA breaks, the arrest of the replication fork, and the realization of the “synthetic lethality” phenomenon due to the inability to repair double-stranded DNA breaks by homologous recombination in cells with mutations in the BRCA1/2 genes. Two randomized trials OlympiAD and EMBRACA evaluated and proved the effectiveness of PARP inhibitors in patients with metastatic BRCA-mutated HER2-negative breast cancer in comparison with standard chemotherapy. At the same time, data on the potential use of PARP inhibitors for the treatment of BRCA-mutated HER2-positive breast cancer patients are extremely limited. This article presents a clinical example of the use of olaparib in a patient with BRCA-mutated HER2-positive metastatic breast cancer

Modern methods for identification and quantification of cardiac glycosides

Scientific relevance. Cardiac glycosides have been used in medicine for over two centuries. Current studies suggest that biologically active substances from this group can be used to treat not only heart conditions but also viral infections, cancers, and other diseases. Therefore, quality control methods for cardiac glycosides are becoming increasingly relevant.Aim. Based on a review of Russian and international quality standards, as well as up-to-date scientific data, this study aimed to identify promising methods for the identification and quantification of cardiac glycosides in herbal drugs and herbal medicinal products, as well as to evaluate the possibility of substituting physicochemical methods for biological methods.Discussion. The methods that are currently used to standardise cardiac glycosides are either not selective or require laboratory animals (biological test systems). According to a study of pharmacopoeial methods for the identification of cardiac glycosides in herbal drugs and herbal medicinal products, chemical identification tests and thin-layer chromatography continue to be relevant. Quantitative testing of herbal drugs and extracts uses biological and non-selective (spectrophotometry) methods, whereas chromatography is described only in general and individual monographs for herbal drug preparations containing individual cardiac glycosides and medicinal products containing these preparations. Upon analysing quality standards and scientific publications, the authors identified potentially promising methods for the quantification of cardiac glycosides in herbal drugs, herbal drug preparations, and herbal medicinal products, namely chromatographic methods.Conclusions. Reverse-phase high-performance liquid cjromatography (HPLC) with spectrophotometric detection is the most suitable method for pharmacopoeial analysis. The development of an HPLC-based analytical procedure to determine the cardiac glycoside content will provide an opportunity to advance from biological or non-selective methods to more ethical and selective up-to-date techniques

Cytoplasmic genetic diversity of potato varieties bred in Russia and FSU countries

Male sterility in potato is little studied since traditional breeding is based  on the vegetative reproduction of highly heterozygous tetraploid varieties. The rapid development of hybrid diploid breeding contributes to growing interest in studying the male sterility of this important crop. In this work, a set of 6 cytoplasmic markers was employed to describe cytoplasmic genetic diversity of 185 potato cultivars bred in Russia and FSU countries. Three cytoplasm types were identified, T (40.0 %), D (50.8 %) and W/γ (8.7 %), which according to literature  are associated with male sterility. With a single exception (0.5 %), cytoplasm types characteristic of male fertile forms (A, P) were not found in the subset  of 185 cultivars. A comparison of these results with previously published data suggested expanding the subset  to up to 277 cultivars, all developed in Russia or FSU countries;  however,  the resulting  differentiation into three cytoplasm  types (T, D and W/γ) was nearly the same. Fertility phenotyping helped identify both  male-sterile and male-fertile genotypes within the three groups  of varieties with T-, D- and W/γ-type cytoplasm. Fifteen genotypes differing in cytoplasm  type and male sterility/fertility traits were selected for direct sequencing of 8 mtDNA loci. Fragments of the  nad2, nad7, cox2, atp6 and  CcmFc genes  were identical  in all 15 selected genotypes. The polymorphism, detected in the rps3, atp9 and CcmFc loci, was not associated with male sterility. Two SNPs in the nad1/atp6 and nad2 loci differentiated 7 genotypes with W/γ-type cytoplasm into five genotypes with tetrad sterility, and two with fertile pollen. The results of an NGS analysis confirmed  the association of these  SNPs with tetrad sterility in a larger set of 28 genotypes of different origin, all with W/γ-type cytoplasm.  A heteroplasmy state  was observed both in male-sterile and in male-fertile genotypes

MOLECULAR SCREENING OF POTATO VARIETIES BRED IN THE NORTHWESTERN ZONE OF THE RUSSIAN FEDERАTION

Increasing the resistance of varieties to diseases and pests is a priority in potato breeding. Many potato varieties created in the Leningrad Scientific Research Institute of Agriculture “Belogorka” are multi-species hybrids possessing complex resistance to the most spread pathogens in combination with a high plasticity, which determined their zoning in many regions of the Russian Federation. Potato varieties and hybrids created in different periods of time in this institute and in the Ltd Breeding company “LiGa” were included in marker-assisted selection with 22 markers of 14 R genes conferring extreme resistance to viruses PVY and PVX, resistance to cyst nematodes Globodera pallida (patotypes Pa2, Pa3) and G. rostochiensis (pathotype Ro1) and resistance to late blight. The data of molecular screening were compared with the results of the State Test of Varieties for Disease Resistance, as well as with other published sources. Markers of sterile and fertile cytoplasm types were also used in the study. Based on the results of molecular screening, varieties and hybrids with different combinations of markers of R genes conferring resistance to the most harmful pathogens (Rysto, Ry-fsto, Rx1, Rx2, Gro1-4, H1, Gpa2, R1, R3a, Rpi-blb1, Rpi-sto1) were detected. Among them, genotypes with sterile D-, T-, W/gamma cytoplasmic types were selected. Analysis of their pollen fertility, crossability and pedigree data showed that breeders succeed in obtaining fertile forms with the D- and T-types of cytoplasm carrying genetic material of Mexican polyploid species Solanum stoloniferum and S. demissum. The information about the presence of markers of R genes and about cytoplasm types of hybrids and varieties will allow an effective implementation of further breeding programs for resistance to a complex of pathogens, including pyramiding of R genes. The data of molecular screening made it possible to specify the pedigrees of several varieties and hybrids that have been created over several decades