Structural Analysis of Glutaredoxin Domain of Mus Musculus Thioredoxin Glutathione Reductase

Thioredoxin glutathione reductase (TGR) is a member of the mammalian thioredoxin reductase family that has a monothiol glutaredoxin (Grx) domain attached to the thioredoxin reductase module. Here, we report a structure of the Grx domain of mouse TGR, determined through high resolution NMR spectroscopy to the final backbone RMSD value of 0.48±0.10 Å. The structure represents a sandwich-like molecule composed of a four stranded β-sheet flanked by five α–helixes, with the CxxS active motif located on the catalytic loop. We structurally characterized the glutathione-binding site in the protein and describe sequence and structural relationships of the domain with glutaredoxins. The structure illuminates a key functional center that evolved in mammalian TGRs to act in thiol-disulfide reactions. Our study allows us to hypothesize that Cys105 might be functionally relevant for TGR catalysis. In addition, the data suggest that the N-terminus of Grx acts as a possible regulatory signal also protecting the protein active site from unwanted interactions in cellular cytosol

NMR approach for monitoring post-mortem changes in Atlantic salmon fillets stored at 0 and 4°C

High resolution NMR technique has been used to monitor post-mortem changes in salmon (Salmo salar) fillets upon storage at 4 and 0 °C. Thirty-one different fish metabolites influencing freshness and taste properties have been unequivocally assigned by NMR using either available standard compounds or ad hoc acquired 2D 1H–1H TOCSY and 1H–13С HSQC spectra. The monitored fish metabolites include amino acids, dipeptides, sugars, vitamins, biogenic amines, as well as different products of the ATP degradation. The detection and monitoring of biogenic amines by NMR, upon fish storage, is information of interest for consumers, since some of these compounds are toxic. The data from this study shows that NMR spectroscopy also provides the amount of all metabolites necessary for the calculation of the K-index used to express fish freshness. A good correlation was found between the K-index increase and the formation of the undesired biogenic amines. The metabolite concentrations and the K-index found in this work were compared and found coherent with literature data. The performed study reveals the strengths and the suitability of the NMR approach to monitor different biochemical processes occurring during fish storage and qualitatively and quantitatively characterise fish metabolites determining fish quality

Characteristics of fatty acid composition of non-traditional feeds used in modern diets of young mink and its effect on the growth of young

From 2010 to 2015, a short-haired mink from Denmark and Poland was imported to the Savvatievo Animal Farm, which is large in size compared to the domestic one. To preserve and further increase the size of the mink, in addition to breeding work, the feeding standards were revised. Since 2010, new non-traditional feeds have been introduced into the diet of animals, the nutritional value of which is not known yet, the caloric content in 2015-2017 was increased to 165-170 kcal and by 2018-2021 it was brought to 180 kcal per 100 g of feed, while at the same time gradually reducing the amount of protein: in 2015-2017 to 7.5 g and in 2018-2021 to 6.5-7.0 g per 100 kcal of feed for young animals during the growth period (July-October). Feed with a high fat content is introduced. In this regard, it was decided to conduct a detailed analysis of some non-traditional feeds by fatty acid composition and determine the effect of the introduction of these feeds on the growth of young animals

Voluntary Refusal to Commit a Crime: Significance, General and Special Signs

The article describes and examines the significance of the rule of the voluntary refusal to commit a crime, as well as explores its general and special signs. It is noted that the voluntary refusal to commit a crime is the rule contained in any modern, progressive law. In this vein, there are different theoretical approaches to the determination of its value and signs. The signs are debatable in nature, and their establishment by the law enforcer may cause difficulties. The difference between voluntary refusal to commit a crime, which is implemented in three functions, is determined, definitions of general signs of voluntary refusal are proposed, their content is clarified. Special signs of voluntary refusal are disclosed

Structural Insights into Interaction between Mammalian Methionine Sulfoxide Reductase B1 and Thioredoxin

Maintenance of the cellular redox balance has vital importance for correcting organism functioning. Methionine sulfoxide reductases (Msrs) are among the key members of the cellular antioxidant defence system. To work properly, methionine sulfoxide reductases need to be reduced by their biological partner, thioredoxin (Trx). This process, according to the available kinetic data, represents the slowest step in the Msrs catalytic cycle. In the present paper, we investigated structural aspects of the intermolecular complex formation between mammalian MsrB1 and Trx. NMR spectroscopy and biocomputing were the two mostly used through the research approaches. The formation of NMR detectable MsrB1/Trx complex was monitored and studied in attempt to understand MsrB1 reduction mechanism. Using NMR data, molecular mechanics, protein docking, and molecular dynamics simulations, it was found that intermediate MsrB1/Trx complex is stabilized by interprotein β-layer. The complex formation accompanied by distortion of disulfide bond within MsrB1 facilitates the reduction of oxidized MsrB1 as it is evidenced by the obtained data

Усовершенствование методов создания гибридов капусты белокочанной

Relevance One of the basic directions of the cabbage crop breeding is the creation of F1 hybrids with a complex of economically valuable traits. This process is difficult and time-consuming as to get pure lines must be within 6-12 years hold inbreeding. Herewith not every line gives the desired heterotic effect that also requires additional verification. Methods Biotechnological method culture of isolated microspores in vitro, which allows in the first generation to receive a line with 100% homozygosity, was used to speed up the breeding process. Combination ability were performed in complete diallel cross on the basic morphological signs. Results Culture medium for cultivation of isolated microspores in vitro was optimized for each genotype of cabbage for the best embryoids regeneration. Maximum amount of embryoids was received on medium with pH 6.2 using ampicillin 100 mg/l and zeatin 1 mg/l: 466.7 ± 153.2 PCs/100 buds. A new source material for breeding – doubled haploid lines of cabbage was received. Lines – the best parents for F1 hybrids with high yield, compact rosette of leaves, with optimum inside and short outside cabbage stump was created. Studies have shown that optimization of breeding process in case of creation of pure lines of cabbage in 3 years with microspore culture requires to reduce the breeding process in 2 times.Актуальность Одним из основных направлений селекции капустных культур является создание гетерозисных гибридов F1 с комплексом хозяйственно ценных признаков. Этот процесс является длительным и трудоемким, поскольку для получения выровненных линий необходимо в течение 6-12 лет проводить инбридинг. При этом не каждая линия дает желаемый гетерозисный эффект, что, в свою очередь, требует дополнительной проверки. Методика Для ускорения некоторых этапов селекционного процесса в работе использовали биотехнологический метод культуры изолированных микроспор in vitro, который позволяет уже в первом поколении получать линии со 100% гомозиготностью. Комбинационную способность определяли в системе полных диаллельных скрещиваний по основным морфологическим признакам. Результаты В результате работы оптимизирован состав питательной среды культуры изолированных микроспор in vitro и индивидуально подобран для каждого генотипа капусты белокочанной для наибольшего выхода эмбриоидов. Максимальный выход эмбриоидов был получен на среде с рН 6,2 с использованием ампициллина 100 мг/л и зеатина 1 мг/л и составил 466,7±153,2 шт./100 бутонов. Получен принципиально новый исходный материал для селекции – удвоенные гаплоидные линии капусты белокочанной. Выделены линии, являющиеся наилучшими исходными формами при создании высокогетерозисных гибридов F1 по урожайности, линии с компактной розеткой листьев, с оптимальной наружной и короткой внутренней кочерыгой. Проведенные исследования показали, что с учетом оптимизации некоторых этапов селекционного процесса для создания чистых линий капусты белокочанной требуется 3 года, что сокращает селекционный процесс в 2 раза

Assessment of Astaxanthin Accumulation in Hepatocytes of Atlantic Salmon Fed Different Diets Using NMR Spectroscopy

This study aimed to assess the astaxanthin (Ax) accumulation in hepatocytes isolated from farmed Atlantic salmon fed different diets (rich marine, poor, poor with marine phospholipids (MPL) and poor with docosahexaenoic acid (DHA)). Nuclear magnetic resonance (NMR) spectroscopy was used for the Ax detection and quantification. The use of the 13C-enriched Ax allowed the assessment of short-time Ax metabolism. The substitution of fish oil and meal in fish feed on plant analogs and the addition of MPL caused further catabolism and decrease of Ax accumulation in hepatocytes from 17 to about 6 mg/kg or to almost zero in the case of DHA addition. Signals assignment of the native and 13C-enriched astaxanthin in acetone were performed using 1D and 2D NMR spectra.publishedVersio