Using the synthetic form RS5 to obtain new introgressive lines of common wheat

The use of the gene pool of wild relatives, which have a significant reserve of genetic diversity, is of immediate interest for breeding common wheat. The creation and use of synthetic forms as “bridges” is an effective method of transferring valuable genetic material from wild relatives to cultivated wheat. For this purpose, genome addition, genome substitution and recombinant “secondary” synthetic forms have been created in the P.P. Lukyanenko National Center of Grain. The synthetic recombination form RS5 (BBAASDt ), in which the third genome consists of chromosomes of Aegilops speltoides (S) and Aegilops tauschii (Dt ), was obtained from crossing the synthetic forms Avrodes (BBAASS) and M.it./Ae. tauschii (BBAADt Dt ), in which the D genome from Ae. tauschii was added to the BBAA genomes of the durum wheat cultivar Mutico italicum. Introgression lines resistant to leaf rust, yellow rust and powdery mildew have been obtained from backcrosses with the susceptible common wheat cultivars Krasnodarskaya 99, Rostislav and Zhirovka. Twelve resistant lines that additionally have high technological characteristics of grain and flour have been selected. The cytological study (С-banding) has revealed chromosomal modifications in 6 of 8 lines under study. The rearrangements mainly affected the chromosomes of the D genome, 1D, 3D, 4D, 6D and 7D. It was found that in most cases the genetic material from the synthetic form RS5 in the studied lines was represented by substituted chromosomes from Ae. tauschii. In line 5791p17, the substitution of chromosomes 6D from Ae. tauschii and 7D from Ae. speltoides was revealed. Substitutions 4D(4Dt ), 6D(6Dt ) from Ae. tauschii and 7D(7S) from Ae. speltoides were obtained for the first time. Molecular analysis of 12 lines did not reveal effective leaf rust resistance genes, presumably present in synthetic forms of M.it./Ae. tauschii and Avrodes. It is assumed that the lines may carry previously unidentified genes for fungal disease resistance, in particular for resistance to leaf rust, from Ae. tauschii and Ae. speltoides

Use of a synthetic form Avrodes for transfer of leaf rust resistance from Aegilops speltoides to common wheat

Diploid wild relative of wheat – Aegilops speltoides – is a valuable source of genes for resistance to diseases. The synthetic form Avrodes (BBAASS) was used as a bridge to transfer leaf rust resistance genes from Ae. speltoides to common wheat. Introgression lines obtained from crosses of Avrodes and susceptible common wheat cultivars were evaluated in a field leaf rust nursery. Resistance levels varied from high to moderate. Testing of lines with the use of molecular markers has shown that some lines have the Lr28 and Lr35 genes inherited from synthetic form Avrodes. The majority of resistance lines have not been found to carry these genes. The Lr47 and Lr51 genes were not identified in the Avrodes and introgression lines. The analysis of chromosome pairing in F1 hybrids showed that the transfer of a genetic material from Avrodes to common wheat basically occurs through translocations. Lines with translocations on chromosomes 2D and 5D were identified by C-banding and FISH. The translocations differed in chromosomal location from known leaf resistance genes transferred to common wheat from Ae. speltoides. Hence it was assumed that new genes were introduced into the common wheat genome from Ae. speltoides. Introgression lines have been studied for productivity and technological qualities of grain. Lines AA60n9 and D37n10 combine high resistance to leaf rust with good characteristics of productivity and technological qualities of grain. The received results demonstrate a genetic diversity and a value of the investigated introgression lines for breeding of common wheat

A study of bread wheat lines from crosses with the synthetic form Avrodes in regard to their yellow rust resistance

The genome-substituted synthetic form Avrodes (AABBSS) was used for transferring resistance to yellow rust (Puccinia striiformis f. sp. tritici Eriks.) from Aegilops speltoides Tausch, (2n = 14) to bread wheat. The study involved 24 introgressive lines of bread wheat obtained using the Avrodes form. Yellow rust resistant lines P07-L.02, P07-L.1, P07-L.17, P07-L.43, P07-L.19, AS12-88, AS12-06, AS12-07, AS12- 51, Asp81-21, Asp63-21, Asp053-21, Asp04-21, Asp022-19, Asp023-19 and Asp029-20 were selected and can be used as new donors of disease resistance. The use of differential chromosome staining (C-banding) and fluorescence in situ hybridization (FISH) identified the genetic material of Ae. speltoides transmitted in the form of 5S(5D) chromosome substitution and translocations of T5BS.5BL-5SL, T2DL.2DS-2SS, T5D, as well as translocation of T1BL.1RS from Secale cereale L. The work revealed that the lines with single translocations of T1BL.1RS and T5BS.5BL-5SL were susceptible to yellow rust, while the lines in which the T2DL.2DS-2SS translocation and 5S(5D) substitutions were identified, as well as the lines with translocations of T1BL.1RS, T2DL.2DS-2SS and T5D showed resistance to the disease. Presumably, the selected introgression lines, obtained by means of crosses with Avrodes, may carry new genes or loci for yellow rust resistance

The development and study of common wheat introgression lines derived from the synthetic form RS7

Synthetic recombination form RS7 (BBAAUS), in which the first two genomes, A and B, originate from common wheat, and the third recombinant genome consists of Aegilops speltoides (S) and Ae. umbellulata (U) chromosomes, was obtained from crossing synthetic forms Avrodes (BBAASS) and Avrolata (BBAAUU). Resistant to leaf rust, yellow rust and powdery mildew, introgression lines have been obtained from backcrosses with the susceptible varieties of common wheat Krasnodarskaya 99, Fisht and Rostislav. PCR analysis showed the presence of amplification fragments with marker SCS421 specific for the Lr28 gene in the line 4991n17. The cytological study (С-banding and FISH) of 14 lines has revealed chromosomal modifications in 12 of them. In most cases, the lines carry translocations from Ae. speltoides, which were identified in chromosomes 1D, 2D, 3D, 2B, 4B, 5B and 7B. Also, lines with the substituted chromosomes 1S (1B), 4D (4S), 5D (5S) and 7D (7S) were identified. Lines that have genetic material from Ae. speltoides and Ae umbellulata at once were revealed. In the line 3379n14, translocations in the short arm of chromosome 7D from Ae. umbellulata and chromosomes 5BL, 1DL, 2DL from Ae. speltoides were revealed. The line 4626p16 presumably has a translocation on the long arm of chromosome 2D from Ae. umbellulata and the T7SS.7SL-7DL translocation from Ae. speltoides. The T1DS.1DL-1SL and T3DS.3DL-3SL translocations from Ae. speltoides, and T2DS.2DL-2UL and T7DL.7DS-7US from Ae. umbellulata have been obtained for the first time. These lines may carry previously unidentified disease resistance genes and, in particular, leaf rust resistance genes from Ae. speltoides and Ae. umbellulata

РЕВМАТИЧЕСКИЕ ЗАБОЛЕВАНИЯ И ТУБЕРКУЛЕЗНАЯ ИНФЕКЦИЯ У ДЕТЕЙ И ВЗРОСЛЫХ

The authors summarized publications on the risk to develop tuberculosis during the treatment of rheumatic diseases with genetically engineered biologic drugs. The article describes the approaches to optimization of tuberculosis control activities in patients with rheumatic diseases: immunodiagnostics of latent tuberculosis infection and prevention of tuberculosis disease in the patients receiving genetically engineered biological drugs.Авторы обобщили публикации по риску развития туберкулеза при лечении ревматических заболеваний генно-инженерными биологическими препаратами. Представлены подходы к оптимизации системы противотуберкулезных мероприятий у больных ревматическими заболеваниями: иммунодиагностика латентной туберкулезной инфекции и предупреждение развития туберкулеза у пациентов, получающих генно-инженерные биологические препараты

СРАВНИТЕЛЬНЫЙ АНАЛИЗ БИОЭКВИВАЛЕНТНОСТИ, КЛИНИЧЕСКОЙ ЭФФЕКТИВНОСТИ И БЕЗОПАСНОСТИ ОРИГИНАЛЬНОГО И ГЕНЕРИЧЕСКОГО ЦИКЛОСПОРИНА ПОСЛЕ ТРАНСПЛАНТАЦИИ ПОЧКИ

The aim of the study was the evaluation of pharmacokinetic parameters and clinical efficacy of generic cyclosporine (Ecoral) and Sandimmune Neoral. Materials and methods’. The pharmacokinetic parameters of different cyclosporine formulations. In 197 kidney graft recipients 319 comprehensive pharmacokinetic studies were performed. In 42 patients received in consecutive order original and generic Cyclosporine in the same dosage the complete pharmacokinetic study was perforfomed. AUC calculations based on dosing interval concentration values were fulfilled using linear trapezoidal rule. To evaluate clinical efficacy 235 short pharmacokinetic studies with concentration examination at 0, 1, 2 and 3 hours after taking Cyclosporine (Co, C1 C2 and C3) were performed in patients treated with Neoral (n = 75) or generic cyclosporine (n = 160). Clinical efficacy of generic cyclosporine was estimated by the prevalence of allograft dysfunction and biopsy proved acute rejection episodes as well as by one-years graft survival and events-free survival. The graft survival rate was calculated by Kaplan–Meyer method. Results. At 100–200 ng/ml maintenance concentration (estimated by C0 concentration) pharmacokinetic parameters did not significantly differ according to Cyclosporine formulation in both complete or short pharmacokinetic studies: AUC-4265 vs. 4204 and 3834 vs. 3670 ng/ml/h respectively; (p > 0.05), Cmax (1036 vs 931 and 813 vs 741 ng/ml respectively; (p > 0.05). Allograft dysfunction occurred in 5% of patients subjected to Neoral immunosuppression and in 8% of Equoral recipients (p > 0.05). However biopsy-proven acute rejection as a cause of graft dysfunction was seen only in patients receiving Ecoral (7% vs 0; p < 0.05). One-years graft survival rate did not differ between groups (99% and 94% in generic CyA and Neoral respectively; p > 0.05), whereas events-free graft survival was significantly lower in patients, receiving generic CyA than in Neoral group (88 vs 94% respectively; p = 0,03). Conclusion. Pharmacokinetic studies have shown the absorption profile of generic formulations Equoral do not differ significantly from that of Neoral. Prospective pilot trial demonstrated no difference between one-year graft survival or graft dysfunction rate, but lower eventsfree one-year graft survival as well as the tendency to higher acute rejection rate in patients treated with generics in comparison with those receiving Neoral should be noted. This issue is to be studied further. Цель исследования: сравнение биодоступности оригинального препарата циклоспорина (CyA) Сандиммуна Неорала и воспроизведенного препарата Экорала, их клинической эффективности и частоты побочных эффектов после трансплантации почки. Материалы и методы. Выполнено 319 фармакокинетических исследований у 197 реципиентов трансплантированной почки (ТП). У 42 из них выполнялось полное фармакокинетическое исследование, то есть определялась концентрация в крови СуА до приема и через 1; 1,5; 2; 3; 4; 6; 8 и 10 часов после приема препарата, причем в каждом случае сначала исследование проводилось на фоне Неорала, а затем через 2 недели после конверсии на Экорал в отношении 1:1. У 155 реципиентов было выполнено 235 укороченных фармакокинетических исследований на фоне приема Неорала (в 75 случаях) либо генерического препарата Экорал (у 160 человек) с определением концентрации в крови СуА до приема, через 1, 2 и 3 часа после приема. Оценивались основные фармакокинетические показатели: уровень СyA в крови до приема препарата (Co), пиковая концентрация (Cmax), время достижения пика (Tmax), площадь под фармакокинетичекой кривой «концентрация – время» (AUC). AUC рассчитывалась по методу трапеции (в полном фармакокинетическом исследовании) либо по формуле Gaspari (в укороченном). Для оценки клинической эффективности проанализированы отдаленные (в течение 12 месяцев после исследования) результаты наблюдения с оценкой функции трансплантата, частоты острого отторжения, однолетней выживаемости трансплантата и «бессобытийной» выживаемости, под которой понимали отсутствие признаков развивающейся дисфункции ТП, отторжения, побочных эффектов или смерти реципиента. Результаты. Основные фармакокинетические параметры значимо не различались после приема Неорала и Экорала как в полном (C max 1036 vs 931 нг/мл и AUC 4265 vs. 4204 нг/мл/ч), так и в укороченном исследовании (813 vs 741 нг/мл и 3834 vs. 3670 нг/мл/ч соответственно). Частота дисфункции трансплантата в этих группах также значимо не различалась (8% на Неорале vs 5% на Экорале), однако обратила на себя внимание тенденция к более частому отторжению на фоне Экорала (7% vs 0), что нашло отражение в более низкой «бессобытийной» выживаемости в этой подгруппе реципиентов (94 и 88% соответственно, p < 0,05). При этом выживаемость трансплантатов через год после исследования на фоне Неорала и Экорала была сопоставимой (94 и 99% соответственно). Заключение. Наши данные позволяют предположить, что при совпадении по важнейшим фармакокинетическим параметрам (Cmax, Tmax, AUC) между генериком Экоралом и Сандиммуном возможны некоторые различия по клинической эффективности. Этот вопрос, однако, требует дальнейшего специального изучения.

Excited-State Electronic Structure with Configuration Interaction Singles and Tamm–Dancoff Time-Dependent Density Functional Theory on Graphical Processing Units

Excited-state calculations are implemented in a development version of the GPU-based TeraChem software package using the configuration interaction singles (CIS) and adiabatic linear response Tamm–Dancoff time-dependent density functional theory (TDA-TDDFT) methods. The speedup of the CIS and TDDFT methods using GPU-based electron repulsion integrals and density functional quadrature integration allows full ab initio excited-state calculations on molecules of unprecedented size. CIS/6-31G and TD-BLYP/6-31G benchmark timings are presented for a range of systems, including four generations of oligothiophene dendrimers, photoactive yellow protein (PYP), and the PYP chromophore solvated with 900 quantum mechanical water molecules. The effects of double and single precision integration are discussed, and mixed precision GPU integration is shown to give extremely good numerical accuracy for both CIS and TDDFT excitation energies (excitation energies within 0.0005 eV of extended double precision CPU results)