Adenocarcinoma of the lung with rare insertion mutation in egfr exon 19 that had partial response to gefitinib: a case report

Aim: Classic activating mutations L858R and deletions in exon 19 (19del) in the gene for epidermal growth factor receptor (EGFR) are associated with sensitivity of the non-small cell lung cancer (NSCLC) to therapy with tyrosine kinase inhibitors (TKI). Insertions in EGFR exon 19 (19ins) are rare mutations in NSCLC; response of cases with 19ins to TKI is not well studied. Here we report a case of NSCLC with 19ins in a Russian patient who was treated with gefitinib. We also overview cases of 19ins reported in the literature. Case description: A 48 years old female Russian patient was diagnosed with adenocarcinoma of the lung (T3N2M1, stage IV). Mutation 19ins was detected in the tumor biopsy by fragment analysis and genotyped by Sanger sequencing as p.I744_K745insKIPVAI. Treatment with gefitinib (250 mg/day) resulted in clinical and radiological improvements scored as partial response that lasted 12 months. Conclusion: Treatment with gefitinib of lung adenocarcinoma that carries mutation EGFR 19ins can result in durable response

Estimation of solvate sphere sizes of cation aquacomplexes in salt solution

The problem of determination of sizes of supermolecular formation (a cluster) consisting of the molecules of water coordinated around a central ion has been analytically solved. The obtained values of the cation aquacomplex point out the possibility of exciting their drift by applying asymmetric electric field with frequency in the range from hundreds of Hz to units of kHz on aqueous salt solution. Drift parameters are determined by inertial properties of aquacomplexes that allows dividing them

The use of wheatgrass (<i>Thinopyrum intermedium</i>) in breeding

Wheatgrass (Th. intermedium) has been traditionally used in wheat breeding for obtaining wheat-wheatgrass hybrids and varieties with introgressions of new genes for economically valuable traits. However, in the 1980s in the United States wheatgrass was selected from among perennial plant species as having promise for domestication and the development of dual-purpose varieties for grain (as an alternative to perennial wheat) and hay. The result of this work was the creation of the wheatgrass varieties Kernza (The Land Institute, Kansas) and MN-Clearwater (University of Minnesota, Minnesota). In Omsk State Agrarian University, the variety Sova was developed by mass selection of the most winter-hardy biotypes with their subsequent combination from the population of wheatgrass obtained from The Land Institute. The average grain yield of the variety Sova is 9.2 dt/ha, green mass is 210.0 dt/ ha, and hay is 71.0 dt/ha. Wheatgrass is a crop with a large production potential, beneficial environmental properties, and valuable grain for functional food. Many publications show the advantages of growing the Kernza variety compared to annual crops in reducing groundwater nitrate contamination, increasing soil carbon sequestration, and reducing energy and economic costs. However, breeding programs for domestication of perennial crops are very limited in Russia. This paper presents an overview of main tasks faced by breeders, aimed at enhancing the yield and cultivating wheatgrass efficiency as a perennial grain and fodder crop. To address them, both traditional and modern biotechnological and molecular cytogenetic approaches are used. The most important task is to transfer target genes of Th. intermedium to modern wheat varieties and decrease the level of chromatin carrying undesirable genes of the wild relative. The first consensus map of wheatgrass containing 10,029 markers was obtained, which is important for searching for genes and their introgressions to the wheat genome. The results of research on the nutritional and technological properties of wheatgrass grain for the development of food products as well as the differences in the quality of wheatgrass grain and wheat grain are presented

The problems of utilizing graphite of stopped graphite-uranium reactors

A list of radioactive nuclides, the activity of which forms the main part of total activity of graphite stack and graphite elements of the construction of stopped industrial graphite-uranium reactors has been defined. The analysis of activity part contributed by these nuclides at different moments of time after stopping reactor was carried out. A set of construction graphite elements, in which there is a possibility of self-sustaining release of the energy stored (Wigner's energy) was determined. It was stated that the most value of the Wigner's energy is achieved in graphite constructions operated in low-temperature region or at high values of flux densities of damaging neutrons and concurrent gamma radiation

Gamma-spectrometric control method of activity and nuclide composition of low-active solid radioactive waste

The gamma-spectrometric control method of low-active solid radioactive waste, based on direct measurement of activity and nuclide composition has been developed. The measurements were carried out in the geometry of standard steel container of 200 l. volume, where low-active wastes were placed. To take into account distribution non-homogeneities of solid waste over the geometry measured a special rotating platform was used, the low-active radioactive wastes being placed on it. Metrological certification was performed and the main errors of this method for 95 % of confidence probability were determined

Electrophysical methods of separation of metal cations in the moving salts solution

The results of experiments on the excitation of the phenomenon of selective drift of solvated ions under the influence of an external "asymmetric" electric field to the circulating solution of calcium chloride and magnesium salts in a polar liquid dielectric - water are shown. The purpose of the experiments was to determine the influence of the field frequency and amplitude of the field strength on the excitation phenomenon, and the study of the operating characteristics of the testing apparatus - a dividing cell. The dependences of the separation efficiency of solvated cations from the frequency of the external field and the excitation threshold of the phenomenon from the field strength in the separation cell are defined

The gene Sr38 for bread wheat breeding in Western Siberia

Present-day wheat breeding for immunity exploits extensively closely related species from the family Triticeae as gene donors. The 2NS/2AS translocation has been introduced into the genome of the cultivated cereal Triticum aestivum from the wild relative T. ventricosum. It contains the Lr37, Yr17, and Sr38 genes, which support seedling resistance to the pathogens Puccinia triticina Eriks., P. striiformis West. f. sp. tritici, and P. graminis Pers. f. sp. tritici Eriks. &amp; E. Henn, which cause brown, yellow, and stem rust of wheat, respectively. This translocation is present in the varieties Trident, Madsen, and Rendezvous grown worldwide and in the Russian varieties Morozko, Svarog, Graf, Marquis, and Homer bred in southern regions. However, the Sr38 gene has not yet been introduced into commercial varieties in West Siberia; thus, it remains of practical importance for breeding in areas where populations of P. graminis f. sp. tritici are represented by avirulent clones. The main goal of this work was to analyze the frequency of clones (a)virulent to the Sr38 gene in an extended West Siberian collection of stem rust agent isolates. In 2019–2020, 139 single pustule isolates of P. graminis f. sp. tritici were obtained on seedlings of the standard susceptible cultivar Khakasskaya in an environmentally controlled laboratory (Institute of Cytology and Genetics SB RAS) from samples of urediniospores collected on commercial and experimental bread wheat fields in the Novosibirsk, Omsk, Altai, and Krasnoyarsk regions. By inoculating test wheat genotypes carrying Sr38 (VPM1 and Trident), variations in the purity of (a)virulent clones were detected in geographical samples of P. graminis f. sp. tritici. In general, clones avirulent to Sr38 constitute 60 % of the West Siberian fungus population, whereas not a single virulent isolate was detected in the Krasnoyarsk collection. The Russian breeding material was screened for sources of the stem rust resistance gene by using molecular markers specific to the 2NS/2AS translocation. A collection of hybrid lines and varieties of bread spring wheat adapted to West Siberia (Omsk SAU) was analyzed to identify accessions promising for the region. The presence of the gene was postulated by genotyping with specific primers (VENTRIUP-LN2) and phytopathological tests with avirulent clones of the fungus. Dominant Sr38 alleles were identified in Lutescens 12-18, Lutescens 81-17, Lutescens 66-16, Erythrospermum 79/07, 9-31, and 8-26. On the grounds of the composition of the West Siberian P. graminis f. sp. tritici population, the Sr38 gene can be considered a candidate for pyramiding genotypes promising for the Novosibirsk, Altai, and Krasnoyarsk regions

Evaluation of synthetic wheat lines (Triticum durum/Aegilops tausсhii) for vegetative period and resistance to diseases

The growth of the total wheat production and increase of yield stability from year to year are some of the priorities of agriculture in Russia. The yield of commercial varieties significantly diversifies due to huge losses of their potential under the influence of negative biotic and abiotic factors. Increase of resistance to stress factors in the emerging varieties can be achieved by utilizing the diversity of the genetic resources of related wild species and genera in crosses. The results of a phenotypic evaluation of the synthetic hexaploid wheat lines of CIMMYT breeding created by crossing durum wheat varieties from Institute of Breeding and Genetics (Odessa, Ukraine) and variety Pandur from Romania (T. durum Desf., AB genome) with Aegilops (Ae. tausshii Coss., D genome), and also 15 synthetic wheat lines of Kyoto University breeding (Japan) created by crossing durum wheat variety Langdon with different ecological forms Aegilops are presented. Research was performed on the experimental field of Omsk SAU under conditions of southern forest-steppe of West Siberia in 2016. Between synthetics, there was revealed a genotypic difference in the vegetative period duration and resistance to diseases. Lines of hybrid combination Aisberg/Ae.sq.(511) were characterized as the most early-maturity genotypes among the lines studied. The hybrid combinations Ae.sq.(369) with variety Aisberg, Ae.sq.(310) and Ukr-Od 1530, Ae.sq.(223) and Pandur are characterized by complex resistance to fungal diseases. Most of the lines demonstrated high and moderate resistance to rust fungus, severity ranging from 5 to 70 % and severity of powdery mildew being 10–90 %. Lines derived from variety Ukr-Od 1530.94 and accessions Ae. tauschii (392); (629); (1027); (1031) and lines Langdon/Ku-2074; Langdon/Ku-2075; Langdon/Ku-2100; Langdon/Ku-2079 are characterized by complex resistance to powdery mildew, leaf and stem rust. The synthetic lines with a complex of economically valuable traits present interest as an initial material for breeding programs

УСЛОВИЯ ЭФФЕКТИВНОГО ПОДАВЛЕНИЯ ПЦР С ПОМОЩЬЮ LNA-ОЛИГОНУКЛЕОТИДОВ ДЛЯ ПРОСТОЙ И ВЫСОКОЧУВСТВИТЕЛЬНОЙ ДЕТЕКЦИИ СОМАТИЧЕСКИХ МУТАЦИЙ

PCR clamping/wild-type blocking PCR with non-extendable locked nucleic acid (LNA) oligonucleotides is used for sensitive detection of somatic mutations in tumors. Various  versions of the technique use different DNA polymerases and LNA oligonucleotides with and  without additional phosphorothioate modifications. Here we studied requirements for successful  PCR clamping with LNA oligonucleotides and Taq DNA polymerase for analysis of mutations in  KRAS and BRAF genes by means of real-time PCR and Sanger sequencing. We found that  addition of phosphorothioate linkages at the 5’-end of LNA oligonucleotide to protect from 5’- exonuclease activity of Taq DNA polymerase did not improve clamping. For most target  sequences, efficient clamping was observed at melting temperature of LNA oligonucleotide  20‑25°C above annealing/extension temperature of the PCR with a 2-step protocol. Under such  conditions, simple and sensitive detection of mutations in KRAS and BRAF genes was feasible using real-time PCR with TaqMan probes or Sanger sequencing.Специфическое блокирование амплификации аллеля дикого типа в ПЦР с помощью олигонуклеотидов, модифицированных по остатку рибозы (закрытые нуклеиновые кислоты, locked nucleic acids, LNA), используется для высокочувствительной детекции соматических мутаций в опухолях. Описаны различные версии метода анализа мутаций с использованием LNA-олигонуклеотидов как с дополнительной модификацией фосфотиоатными группами, так и без таких групп, при этом использовались  различные ДНК полимеразы. В работе проведен анализ оптимальных условий для успешного специфического блокирования ПЦР с помощью LNA-олигонуклеотидов при анализе мутаций в генах KRAS и  BRAF. Мы обнаружили, что фосфотиоатная защита на 5’-конце олигонуклеотидов не влияет на эффективность блокирования аллеля дикого типа. Выявлено, что для большинства последовательностей  эффективное блокирование наблюдается при проведении шага отжига и элонгации ПЦР при температуре на  20–25°С ниже температуры плавления LNA-олигонуклеотида. При таких условиях реакции возможна простая и высокочувствительная детекция мутаций в генах KRAS и BRAF с использованием как секвенирования по Сэнгеру, так и ПЦР в реальном времени с Taqman зондами