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"Timofeeva, Olga"
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Conditional reprogramming and long-term expansion of normal and tumor cells from human biospecimens
This protocol demonstrates how to establish primary epithelial cell cultures
in vitro
from healthy human tissue and human cancer samples using ROCK inhibitor and irradiated feeder cells.
Historically, it has been difficult to propagate cells
in vitro
that are derived directly from human tumors or healthy tissue. However,
in vitro
preclinical models are essential tools for both the study of basic cancer biology and the promotion of translational research, including drug discovery and drug target identification. This protocol describes conditional reprogramming (CR), which involves coculture of irradiated mouse fibroblast feeder cells with normal and tumor human epithelial cells in the presence of a Rho kinase inhibitor (Y-27632). CR cells can be used for various applications, including regenerative medicine, drug sensitivity testing, gene expression profiling and xenograft studies. The method requires a pathologist to differentiate healthy tissue from tumor tissue, and basic tissue culture skills. The protocol can be used with cells derived from both fresh and cryopreserved tissue samples. As approximately 1 million cells can be generated in 7 d, the technique is directly applicable to diagnostic and predictive medicine. Moreover, the epithelial cells can be propagated indefinitely
in vitro
, yet retain the capacity to become fully differentiated when placed into conditions that mimic their natural environment.
Journal Article
The Art of Dying: Making a Will in Old English and Its Sociolinguistic Context
This paper explores the potential of legal documents for the study of the sociology of Old English. It gives a rationale for the use of legal genres, or charters, and introduces research databases and tools that may elucidate the interconnections between practitioners of legal Old English and their linguistic practices. A series of short case studies on wills illustrates what legal genres tell us about the correlation between linguistic variation, supralocalisation, and change and such variables as archive and gender.
Journal Article
STAT3 suppresses transcription of proapoptotic genes in cancer cells with the involvement of its N-terminal domain
Activation of STAT3 in cancers leads to gene expression promoting cell proliferation and resistance to apoptosis, as well as tumor angiogenesis, invasion, and migration. In the characterization of effects of ST3-H2A2, a selective inhibitor of the STAT3 N-terminal domain (ND), we observed that the compound induced apoptotic death in cancer cells associated with robust activation of proapoptotic genes. Using ChIP and tiling human promoter arrays, we found that activation of gene expression in response to ST3-H2A2 is accompanied by altered STAT3 chromatin binding. Using inhibitors of STAT3 phosphorylation and a dominant-negative STAT3 mutant we found that the unphosphorylated form of STAT3 binds to regulatory regions of proapoptotic genes and prevents their expression in tumor cells but not normal cells. siRNA knockdown confirmed the effects of ST3-HA2A on gene expression and chromatin binding to be STAT3 dependent. The STAT3-binding region of the C/EBP-homologous protein (CHOP) promoter was found to be localized in DNasel hypersensitive site of chromatin in cancer cells but not in nontransformed cells, suggesting that STAT3 binding and suppressive action can be chromatin structure dependent. These data demonstrate a suppressive role for the STAT3 ND in the regulation of proapoptotic gene expression in cancer cells, providing further support for targeting STAT3 ND for cancer therapy.
Journal Article
Transcutaneous spinal cord stimulation modulates quiet standing in healthy adults: stimulation site and cognitive style matter
The study explored the effects of transcutaneous electrical spinal cord stimulation (tES) on postural control. Subjects were divided into field-dependent (FD) and field-independent (FI) groups according to their cognitive style. FD subjects use an exteroceptive afferent stream for spatial orientation, while FI subjects use an interoceptive stream. In darkness, vertical posture is maintained by head-trunk stabilization in FD subjects and by independent movements of body segments in FI subjects. Previously, we showed that tES at the L1-L2 vertebral level decreased postural stability in FD subjects. Now, stimulation was applied at the T11-T12 vertebral level (midline, above the left or right dorsal roots). Quiet standing was assessed using stabilometry in 18 FD and FI participants. Participants stood on a force platform in soundproof chamber with eyes closed during tES. Midline and left tES significantly improved postural stability by up to 28% in FD participants, while posture did not change significantly in FI participants. Pronounced differences between the effects of T11-T12 and L1-L2 stimulation are associated with selective topographical activation of proximal and distal leg muscles during tES of the lumbar enlargement. This study highlights the importance of considering cognitive style in postural control research.
Journal Article
Sociolinguistic variation in Old English : records of communities and people
The first extensive study of Old English to utilise the insights and methodologies of sociolinguistics. A description of OE speech communities informed by the theory of social networks and communities of practice, with special attention to processes of supralocalisation and their correlation to political centralisation in Anglo-Saxon England.
eBook
Spinal Regulation of Posture: Effects of Transcutaneous Spinal Cord and Affective Sound Stimulation
The combined effects of transcutaneous electrical stimulation (tES) of the spinal cord and affective sound stimulation on postural control were investigated to elucidate the involvement of spinal networks in the maintenance of vertical stability. Healthy volunteers (n = 27) underwent tES and sound stimulation separately and combined quasi-randomly. All participants were field-dependent; i.e., participants used the exteroceptive afferent stream for spatial orientation. Centre-of-pressure parameters were analysed to assess postural stability. Results showed that tES at the T11–T12 vertebrae stabilised posture, tES at the L1–L2 vertebrae had no postural effect, and sound stimulation from the left destabilised posture. To assess the role of spinal regulation of postural disturbances, we compared the effects of combined tES with sound stimulation to those of sound stimulation alone. Stimulation at the T11–T12 level reduced the lateral sway induced by affective sounds, whereas L1–L2 tES did not. These results suggest that, in healthy individuals, spinal networks located at the T11–T12 and L1–L2 vertebral levels have distinct roles in maintaining upright posture, both when a person is standing still and when they are actively stabilising their posture during destabilising perturbations. T11–T12 spinal networks stabilise upright posture when destabilising information is solely transmitted from the supraspinal level.
Journal Article
Posture of Healthy Subjects Modulated by Transcutaneous Spinal Cord Stimulation
Transcutaneous electrical stimulation of the spinal cord is used to restore locomotion and body weight support in patients with severe motor disorders. We studied the effects of this non-invasive stimulation on postural control in healthy subjects. Stimulation at the L1–L2 vertebrae was performed to activate the extensor muscles of the lower limbs. Because postural regulation depends on the cognitive style, the effects of the stimulation were analyzed separately in field-dependent (FD) and field-independent (FI) participants. During the study, FD and FI participants (N = 16, 25 ± 5 years, all right dominant leg) stood on a force platform in a soundproof chamber with their eyes closed. Stimulation was applied in the midline between the L1 and L2 vertebrae or over the left or right dorsal roots of the spinal cord; under the control condition, there was no stimulation. Stimulation destabilized posture in healthy subjects, whereas patients with movement disorders usually showed an improvement in postural control. In the FD participants, left dorsal root and midline stimulation increased several postural parameters by up to 30%. Dorsal root stimulation on the side of the supporting leg reduced postural control, while stimulation on the side of the dominant leg did not. No significant changes were observed in the FI participants.
Journal Article
Bide Nu Æt Gode Þæt Ic Grecisc Cunne: Attitudes to Greek and the Greeks in the Anglo-Saxon Period
The Greeks were one of those outgroups to whom the Anglo-Saxons had reasons to look up to, because of the antiquity of their culture and the sanctity of their language, along those of the Hebrews and the Romans. Yet as a language Greek was practically unknown for most of the Anglo-Saxon period and contact with its native speakers and country extremely limited. Nevertheless, references to the Greeks and their language are not uncommon in the Anglo-Saxon sources (both Latin and vernacular), as a little less than 200 occurrences in the Dictionary of Old English (s.v. grecisc) testify.
This paper uses these data, supplementing them with searches in the Dictionary of Old English Web Corpus, Brepolis Library of Latin Texts - Series A, monumenta.ch and Medieval Latin from Anglo-Saxon Sources, and analyses lexical and syntactic strategies of the Greek outgroup construction in Anglo-Saxon texts. It looks at lexemes denoting ‘Greek’ and their derivatives in Anglo-Latin and Old English, examines their collocates and gleans information on attitudes towards Greek and the Greeks, and on membership claims indexed by Latin-Greek or English-Greek code-switching, by at the same time trying to establish parallels and influences between the two high registers of the Anglo-Saxon period.
Journal Article
Development of an industrial cyber-physical platform for small series production using digital twins
The article describes an industrial cyber-physical platform for small series production using digital twins under development at ITMO University (Saint Petersburg, Russia). The platform is based on the following approaches: group technology, adaptive and selective assembling, and digital twin of production systems and processes. The article presents a mechanism for constructing a unified manufacturing process, and results of an integrated multiscale simulation of an injection moulding process. The issues of ensuring identification and monitoring of objects of the industrial cyber-physical platform are considered. Specific service applications required to implement the smart product concept are discussed. The combination of the considered technologies is used to create digital twins of production system objects. All humans that have different roles in the product value stream can interact with the industrial cyber-physical platform at the three levels, receiving support in performing their tasks. This article is part of the theme issue ‘Towards symbiotic autonomous systems’.
Journal Article