Bioreactor cultivation of CHO DP-12 cells under sodium butyrate treatment – comparative transcriptome analysis with CHO cDNA microarrays

Klausing S, Krämer O, Noll T. Bioreactor cultivation of CHO DP-12 cells under sodium butyrate treatment – comparative transcriptome analysis with CHO cDNA microarrays. BMC Proceedings. 2011;5(Suppl 8)

Doan Ye Cry Ma Honey / music by Albert V. Noll; words by Albert V. Noll

Cover: description reads darky song; Publisher: Oliver Ditson Company (Boston)https://egrove.olemiss.edu/sharris_a/1047/thumbnail.jp

Enhancing stability of recombinant CHO cells by CRISPR/Cas9-mediated site-specific integration into regions with distinct histone modifications

Chinese hamster ovary (CHO) cells are the most important platform for producing biotherapeutics. Random integration of a transgene into epigenetically instable regions of the genome results in silencing of the gene of interest and loss of productivity during upstream processing. Therefore, cost- and time-intensive long-term stability studies must be performed. Site-specific integration into safe harbors is a strategy to overcome these limitations of conventional cell line design. Recent publications predict safe harbors in CHO cells based on omics data sets or by learning from random integrations, but those predictions remain theory. In this study, we established a CRISPR/Cas9-mediated site-specific integration strategy based on ChIP-seq data to improve stability of recombinant CHO cells. Therefore, a ChIP experiment from the exponential and stationary growth phase of a fed-batch cultivation of CHO-K1 cells yielded 709 potentially stable integration sites. The reporter gene eGFP was integrated into three regions harboring specific modifications by CRISPR/Cas9. Targeted Cas9 nanopore sequencing showed site-specific integration in all 3 cell pools with a specificity between 23 and 73%. Subsequently, the cells with the three different integration sites were compared with the randomly integrated donor vector in terms of transcript level, productivity, gene copy numbers and stability. All site-specific integrations showed an increase in productivity and transcript levels of up to 7.4-fold. In a long-term cultivation over 70 generations, two of the site-specific integrations showed a stable productivity (>70%) independent of selection pressure

Doan Ye Cry, Ma Honey / words by Albert W. Noll

Cover: text reads: Songs in Negro Dialect; Publisher: Oliver Ditson Company (Boston)https://egrove.olemiss.edu/sharris_a/1061/thumbnail.jp

Разработка устройства управления беспроводной связью на базе чипа ESP8266

Цель выпускной квалификационной работы – разработка устройства управления беспроводной связью на базе чипа ESP8266.За время выполнения работы проведено ознакомление возможностями элементов используемых в работе, а именно микроконтроллеров на базе ядра ARM-Cortex-M3 и Wi-Fi модулей, выполненных на чипсете ESP8266. В соответствии с требованиями, отраженными в техническом задании разработана структура и выполнен ручной монтаж устройства. Конечным этапом достижения цели являлись разработка алгоритма функционирования устройства и выполнение его программной реализации.Разработанное устройство управления беспроводной связью рекомендовано для использования в системах мониторинга охранно-пожарной, тревожной и технологической сигнализации стационарных объектов.The objective of the graduation thesis is to develop the wireless control device based on ESP8266 chipset. In the course of the work, the author examined the capabilities of the elements used in the work, namely the microcontrollers based on the ARM-Cortex-M3 core and the Wi-Fi modules based on the ESP8266 chipset. In accordance with the requirements set forth in the Statement of Work, the author developed the structure and carried out the manual assembly of the device. The final objective is to develop the operation algorithm of the device and to ensure software implementation of this algorithm.The developed wireless control device is recommended to be used in monitoring systems of fire, security, emergency and process alarm devices for stationary objects

Effect of Bioactive and Antimicrobial Nanoparticles on Properties and Applicability of Dental Adhesives

The aim of the study was to examine the applicability of bioactive and antibacterial nanoparticles to an experimental adhesive. The adhesive (60 wt% BisGMA, 15 wt% TEGDMA, 25 wt% HEMA) was mixed with combinations of 5 wt% methacryl-functionalized polyhedral oligomeric silsesquioxane (MA-POSS) and one kind of bioactive/antibacterial nanoparticles: 1 wt% core-shell silica-silver nanoparticle (SiO2@Ag), 1 wt% bioactive glass with bismuth (BAG-Bi) or 1 wt% calcium phosphate (CAP). Pure adhesive served as control. The physicochemical (degree of conversion (DC), linear shrinkage (LS), shear and complex viscosity, water sorption (WS), sol fraction (SF)), biological (antimicrobial effect) and bioactive (mineral precipitation) properties were investigated. DC and LS remained unchanged. The combination of BAG-Bi/MA-POSS resulted in a significantly increased WS and SF compared to control. In addition, the combination of CAP/MA-POSS slightly increased the shear viscosity of the adhesive. The addition of the nanoparticles did not influence the antimicrobial effects compared to the pure adhesive. Improved mineral inducing capacity could be detected in all nanoparticle combinations. The combination of bioactive and/or antibacterial nanoparticles showed improved mineral inducing capacity, but no antibacterial properties. The material properties were not or only slightly affected. Keywords: POSS; SiO2@Ag; antimicrobial nanoparticles; bioactive glass; bioactive nanoparticles; calcium phosphate; dental adhesive

Adult Palatum as a Novel Source of Neural Crest-Related Stem Cells

Somatic neural and neural crest stem cells are promising sources for cellular therapy of several neurodegenerative diseases. However, because of practical considerations such as inadequate accessibility of the source material, the application of neural crest stem cells is strictly limited. The secondary palate is a highly regenerative and heavily innervated tissue, which develops embryonically under direct contribution of neural crest cells. Here, we describe for the first time the presence of nestin-positive neural crest-related stem cells within Meissner corpuscles and Merkel cell-neurite complexes located in the hard palate of adult Wistar rats. After isolation, palatal neural crest-related stem cells (pNC-SCs) were cultivated in the presence of epidermal growth factor and fibroblast growth factor under serum-free conditions, resulting in large amounts of neurospheres. We used immunocytochemical techniques and reverse transcriptase-polymerase chain reaction to assess the expression profile of pNC-SCs. In addition to the expression of neural crest stem cell markers such as Nestin, Sox2, and p75, we detected the expression of Klf4, Oct4, and c-Myc. pNC-SCs differentiated efficiently into neuronal and glial cells. Finally, we investigated the potential expression of stemness markers within the human palate. We identified expression of stem cell markers nestin and CD133 and the transcription factors needed for reprogramming of somatic cells into pluripotent cells: Sox2, Oct4, Klf4, and c-Myc. These data show that cells isolated from palatal rugae form neurospheres, are highly plastic, and express neural crest stem cell markers. In addition, pNC-SCs may have the ability to differentiate into functional neurons and glial cells, serving as a starting point for therapeutic studies. Stem Cells 2009;27:1899–191