Ice-Cap: A Method for Growing Arabidopsis and Tomato Plants in 96-well Plates for High-Throughput Genotyping

It is becoming common for plant scientists to develop projects that require the genotyping of large numbers of plants. The first step in any genotyping project is to collect a tissue sample from each individual plant. The traditional approach to this task is to sample plants one-at-a-time. If one wishes to genotype hundreds or thousands of individuals, however, using this strategy results in a significant bottleneck in the genotyping pipeline. The Ice-Cap method that we describe here provides a high-throughput solution to this challenge by allowing one scientist to collect tissue from several thousand seedlings in a single day 1,2. This level of throughput is made possible by the fact that tissue is harvested from plants 96-at-a-time, rather than one-at-a-time

ВПЛИВ НЕСТАЦІОНАРНИХ ЕЛЕКТРОМАГНІТНИХ ПРОЦЕСІВ НА ЕЛЕКТРОМЕХАНІЧНІ ПАРАМЕТРИ У СИНХРОННОМУ ЕЛЕКТРОПРИВОДІ

Purpose. Investigation of the influence of sharply variable electric load on non-stationary electromagnetic phenomena and electromechanical processes in a synchronous electric drive.Methodology. Theoretical and experimental methods of research and analysis of a set of parameters of sharply varying loads of electrotechnological complexes and electromagnetic processes in electrical equipment; physical and analytical methods for obtaining dependencies of the influence of electrotechnical loads on the technical condition, the breakdown of electrical equipment, additional losses and non-stationary electromagnetic phenomena in systems and equipment.Findings. The calculations of the excess of the oscillatory processes of the synchronous electric drive shaft are performed, which are excited by sharply varying loads, relative to the norms standardized by state standards. Proceeding from the primary requirements of the electric drive and providing normalized oscillations of the motor shaft and increasing requirements to reliability, efficiency and energy consumption per unit of output, the effect of non-stationary electromagnetic processes on the electromechanical parameters of the electric drive was investigated.Originality.  A mathematical technique is proposed for non-stationary electromagnetic processes in a synchronous electric drive, which allows one to investigate the effect of non-sinusoidal and unbalanced currents and voltages on electromechanical parameters. An engineering technique for estimating the oscillatory function of a shaft is given, depending on the nature of non-stationary electromagnetic processes in a synchronous motor.Practical value.  An engineering technique for estimating the oscillations of a synchronous electric drive shaft is proposed for non-stationary electromagnetic processes, which lead to an increase in the parameters of normative and technical documentation.Цель работы. Исследование влияния резкопеременной электрической нагрузки на нестационарные электромагнитные явления и электромеханические процессы в синхронном электроприводе.Методы исследования. Теоретические и экспериментальные методики исследований и анализа совокупности параметров резкопеременных нагрузок электротехнологических комплексов и электромагнитных процессов в электрооборудовании; физико-аналитические методики для получения зависимостей влияния электротехнологических нагрузок на техническое состояние, аварийность электрооборудования, добавочные потери и нестационарных электромагнитных явлений в системах и оборудовании.Полученные результаты. Выполнены расчеты превышения колебательных процессов вала синхронного электропривода, которые возбуждаются резкопеременными нагрузками, относительно нормированных государственными стандартами. Исходя из первостепенных требований электропривода и обеспечения нормируемых колебаний вала двигателя и возрастающих требований к надежности, эффективности и снижения энергозатрат на единицу вырабатываемой продукции было исследовано влияние нестационарных электромагнитных процессов на электромеханические параметры электропривода.Научна новизна. Предложена математическая методика нестационарных электромагнитных процессов в синхронном электроприводе, которая позволяет исследовать влияние несинусоидальности и несимметрии токов и напряжений на электромеханические параметры. Приведена инженерная методика оценки колебательной функции вала в зависимости от характера нестационарных электромагнитных процессов в синхронном электродвигателе.Практическая ценность. Предложена инженерная методика оценки колебаний вала синхронного электропривода при нестационарных электромагнитных процессах, приводящих к увеличению показателей нормативно-технической документации.Мета роботи. Дослідження впливу різкозмінного електричного навантаження на нестаціонарні електромагнітні явища та електромеханічні процеси у синхронному електроприводі.Методи дослідження. Теоретичні та експериментальні методики дослідження і аналізу сукупності параметрів різкозмінних навантажень електротехнологічних комплексів і електромагнітних процесів в електрообладнанні; фізико-аналітичні методики для отримання залежності впливу електротехнологічних навантажень на технічний стан, аварійність електрообладнання, додаткові втрати та нестаціонарних електромагнітних явищ в системах і обладнанні.Отримані результати. Виконано розрахунки перевищення коливальних процесів валу синхронного електроприводу, які збуджуються різкозмінними навантаженнями, відносно нормованих державними стандартами. Виходячи з першорядних вимог електроприводу і забезпечення нормованих коливань валу двигуна і зростаючих вимог до надійності, ефективності та зниження енерговитрат на одиницю вироблюваної продукції було досліджено вплив нестаціонарних електромагнітних процесів на електромеханічні параметри електроприводу.Наукова новизна. Запропоновано математичну методику нестаціонарних електромагнітних процесів в синхронному електроприводі, яка дозволяє досліджувати вплив несинусоїдальності і несиметрії струмів і напруг на електромеханічні параметри. Наведено інженерну методику оцінки коливальної функції валу залежно від характеру нестаціонарних електромагнітних процесів в синхронному електродвигуні.Практична цінність. Запропоновано інженерну методику оцінки коливань валу синхронного електроприводу при нестаціонарних електромагнітних процесах, що призводять до збільшення показників нормативно-технічної документації.

Chronic IL-1β-induced inflammation regulates epithelial-to-mesenchymal transition memory phenotypes via epigenetic modifications in non-small cell lung cancer.

Chronic inflammation facilitates tumor progression. We discovered that a subset of non-small cell lung cancer cells underwent a gradually progressing epithelial-to-mesenchymal (EMT) phenotype following a 21-day exposure to IL-1β, an abundant proinflammatory cytokine in the at-risk for lung cancer pulmonary and the lung tumor microenvironments. Pathway analysis of the gene expression profile and in vitro functional studies revealed that the EMT and EMT-associated phenotypes, including enhanced cell invasion, PD-L1 upregulation, and chemoresistance, were sustained in the absence of continuous IL-1β exposure. We referred to this phenomenon as EMT memory. Utilizing a doxycycline-controlled SLUG expression system, we found that high expression of the transcription factor SLUG was indispensable for the establishment of EMT memory. High SLUG expression in tumors of lung cancer patients was associated with poor survival. Chemical or genetic inhibition of SLUG upregulation prevented EMT following the acute IL-1β exposure but did not reverse EMT memory. Chromatin immunoprecipitation and methylation-specific PCR further revealed a SLUG-mediated temporal regulation of epigenetic modifications, including accumulation of H3K27, H3K9, and DNA methylation, in the CDH1 (E-cadherin) promoter following the chronic IL-1β exposure. Chemical inhibition of DNA methylation not only restored E-cadherin expression in EMT memory, but also primed cells for chemotherapy-induced apoptosis

Immunolocalization of dually phosphorylated MAPKs in dividing root meristem cells of Vicia faba, Pisum sativum, Lupinus luteus and Lycopersicon esculentum

Key message In plants, phosphorylated MAPKs display constitutive nuclear localization; however, not all studied plant species show co-localization of activated MAPKs to mitotic microtubules. Abstract The mitogen-activated protein kinase (MAPK) signaling pathway is involved not only in the cellular response to biotic and abiotic stress but also in the regulation of cell cycle and plant development. The role of MAPKs in the formation of a mitotic spindle has been widely studied and the MAPK signaling pathway was found to be indispensable for the unperturbed course of cell division. Here we show cellular localization of activated MAPKs (dually phosphorylated at their TXY motifs) in both interphase and mitotic root meristem cells of Lupinus luteus, Pisum sativum, Vicia faba (Fabaceae) and Lycopersicon esculentum (Solanaceae). Nuclear localization of activated MAPKs has been found in all species. Colocalization of these kinases to mitotic microtubules was most evident in L. esculentum, while only about 50 % of mitotic cells in the root meristems of P. sativum and V. faba displayed activated MAPKs localized to microtubules during mitosis. Unexpectedly, no evident immunofluorescence signals at spindle microtubules and phragmoplast were noted in L. luteus. Considering immunocytochemical analyses and studies on the impact of FR180204 (an inhibitor of animal ERK1/2) on mitotic cells, we hypothesize that MAPKs may not play prominent role in the regulation of microtubule dynamics in all plant species

Tumor Response to Combination Celecoxib and Erlotinib Therapy in Non-small Cell Lung Cancer Is Associated with a Low Baseline Matrix Metalloproteinase-9 and a Decline in Serum-Soluble E-Cadherin

IntroductionCyclooxygenase-2 overexpression may mediate resistance to epidermal growth factor receptor tyrosine kinase inhibition through prostaglandin E2-dependent promotion of epithelial to mesenchymal transition (EMT). Suppression of epithelial markers, such as E-cadherin, can lead to resistance to erlotinib. Prostaglandin E2 down-regulates E-cadherin expression by up-regulating transcriptional repressors, including ZEB1 and Snail. Furthermore, E-cadherin can be modulated by matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs), promoting tumor invasion and metastasis. Markers of EMT and tumor invasion were evaluated in patient serum from a phase I clinical trial investigating the combination of celecoxib and erlotinib in non-small cell lung cancer (NSCLC) patients.MethodsSamples from 22 subjects were evaluated. Soluble E-cadherin (sEC) was evaluated by enzyme linked immunosorbent assay in patient serum at baseline, week 4, and week 8 of treatment. Other markers of EMT and angiogenesis were evaluated by enzyme linked immunosorbent assay, including MMP-9, TIMP-1, and CCL15.ResultsSerum sEC, MMP-9, TIMP-1, and CCL15 levels were determined at baseline and week 8. Patients with a partial response to therapy had a significant decrease in sEC, TIMP-1, and CCL15 at week 8. In patients who responded to the combination therapy, baseline MMP-9 was significantly lower compared with nonresponders (p = 0.006).ConclusionssEC, MMP-9, TIMP-1, and CCL15 levels correlate with response to combination therapy with erlotinib and celecoxib in patients with NSCLC. A randomized phase II trial is planned comparing erlotinib and celecoxib with erlotinib plus placebo in advanced NSCLC. This study will prospectively assess these and other biomarkers in serum and tumor tissue

Differences in proteolytic activity and gene profiles of fungal strains isolated from the total parenteral nutrition patients

Fungal infections constitute a serious clinical problem in the group of patients receiving total parenteral nutrition. The majority of species isolated from infections of the total parenteral nutrition patients belong to Candida genus. The most important factors of Candida spp. virulence are the phenomenon of “phenotypic switching,” adhesins, dimorphism of fungal cells and the secretion of hydrolytic enzymes such as proteinases and lipases, including aspartyl proteinases. We determined the proteolytic activity of yeast-like fungal strains cultured from the clinical materials of patients receiving total parenteral nutrition and detected genes encoding aspartyl proteinases in predominant species Candida glabrata—YPS2, YPS4, and YPS6, and Candida albicans—SAP1–3, SAP4, SAP5, and SAP6. C. albicans released proteinases on the various activity levels. All C. glabrata strains obtained from the clinical materials of examined and control groups exhibited secretion of the proteinases. All 13 isolates of C. albicans possessed genes SAP1–3. Gene SAP4 was detected in genome of 11 C. albicans strains, SAP5 in 6, and SAP6 in 11. Twenty-six among 31 of C. glabrata isolates contained YPS2 gene, 21 the YPS4 gene, and 28 the YPS6 gene. We observed that clinical isolates of C. albicans and C. glabrata differed in SAPs and YPSs gene profiles, respectively, and displayed differentiated proteolytic activity. We suppose that different sets of aspartyl proteinases genes as well as various proteinase-activity levels would have the influence on strains virulence

Secondary contact and admixture between independently invading populations of the Western corn rootworm, diabrotica virgifera virgifera in Europe

The western corn rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), is one of the most destructive pests of corn in North America and is currently invading Europe. The two major invasive outbreaks of rootworm in Europe have occurred, in North-West Italy and in Central and South-Eastern Europe. These two outbreaks originated from independent introductions from North America. Secondary contact probably occurred in North Italy between these two outbreaks, in 2008. We used 13 microsatellite markers to conduct a population genetics study, to demonstrate that this geographic contact resulted in a zone of admixture in the Italian region of Veneto. We show that i) genetic variation is greater in the contact zone than in the parental outbreaks; ii) several signs of admixture were detected in some Venetian samples, in a Bayesian analysis of the population structure and in an approximate Bayesian computation analysis of historical scenarios and, finally, iii) allelic frequency clines were observed at microsatellite loci. The contact between the invasive outbreaks in North-West Italy and Central and South-Eastern Europe resulted in a zone of admixture, with particular characteristics. The evolutionary implications of the existence of a zone of admixture in Northern Italy and their possible impact on the invasion success of the western corn rootworm are discussed